Table of Contents2023.11.05 Nucleic Acids - DNA TSP 70........................................................................................12023.11.07 Bacteriophage..........................................................................................................22023.11.07 Precipitate Comparisons...........................................................................................32023.11.12 Decanted - Vinyl Production?.....................................................................................42023.11.14 TSP Culture Variants.................................................................................................62023.11.14 TSP culture variation................................................................................................92023.11.15 TSP Culture Variations............................................................................................102023.11.16 (Borax Polymer) & 100.6.........................................................................................122023.11.17 Solubility Methylation.............................................................................................162023.11.18 Borax polymer solubility..........................................................................................192023.11.19 Project Planning.....................................................................................................232023.11.20 UV NIR TSP 70 Nucleic Acid......................................................................................252023.11.21 Nucleic Acids (?) - DNA- PVA....................................................................................272023.11.22 Solubility-Health-Blood(Syn)....................................................................................312023.11.23 Blood(Syn)-Polymer Compare..................................................................................372023.11.24 Abstracts AI - NIR Synth..........................................................................................432023.11.25 Human Synth Blood Compare..................................................................................462023.11.26 Blood Synth Vinyl MID IR.........................................................................................502023.11.27 Research Paper:Blood Compare...............................................................................542023.11.28 Research Paper Video Audio....................................................................................572023.11.30 Another Mitigation Prospect....................................................................................602023.12.02 Abstracts 2022 Prepared.........................................................................................662023.12.04 Media Summaries - Vinyl.........................................................................................672023.12.05 Blood Clot Progession Study....................................................................................792023.12.06 Functional Group Worksheet...................................................................................812023.12.07 Functional Group Worksheet...................................................................................832023.12.08 Functional Group Worksheet...................................................................................952023.12.09 Functional Groups DNA...........................................................................................992023.12.10 Functional Group Worksheet..................................................................................1012023.12.11 Functional Group Worksheet..................................................................................1042023.12.12 Research Paper - Regroup.....................................................................................1072023.12.13 Alkenes - Polymerization.......................................................................................1172023.12.14 Mitigation -Methylation.........................................................................................118I
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Table of Contents2023.12.15 Paper Complete....................................................................................................1262023.12.16 Bacteriophage Inquiry...........................................................................................1282023.12.17 Enzyme - Bacteriophage - DNA...............................................................................1322023.12.18 DNA - Bacteriophage Inquiry..................................................................................1422023.12.19 DNA - Bacteriophage Inquiry..................................................................................1512023.12.23 DNA - Bacteriophage Inquiry..................................................................................1552023.12.24 Video Up - Regroup - Phage...................................................................................1582023.12.25 Christmas - On We Go............................................................................................1642023.12.26 Five TSP Separations.............................................................................................1662023.12.27 Amines Amides Vinyl.............................................................................................1692023.12.28 TSP 070 Separations LC DNA.................................................................................1722023.12.29 TSP 070 Acetone Study..........................................................................................1742023.12.30 TSP 070 Acetone-Phage-ImageDB..........................................................................1772023.12.31 Afflicted Skin........................................................................................................179II
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2023.11.05 Nucleic Acids - DNA TSP 70NucleotidesNucleic AcidsDNA?2023.11.05 Nucleic Acids - DNA TSP 701 2023.11.07 BacteriophageBacteriophage issue arises today.2023.11.07 Bacteriophage2 2023.11.07 Precipitate ComparisonsCompared NIR plotsNative Preciptate 20 Days (70.5) vs Titrated Precip (70.2)70.5 > 70.2:1195(Methyl) > 933(Methylene)1506 Aromatic Amine, NH Amide Protein > 1515 Polyamide1582 Aklyl Alcohol > 1611-1654 Vinyl, Polyamide, Acrylate2023.11.07 Precipitate Comparisons3 2023.11.12 Decanted - Vinyl Production?From UV analysis:TyrosineProteinStrong surface polymer forming. Grid like structure under slide. CDB polymer grid matrix.No definite VIS NIR. This does say that the alkyl alcohol is no longer in place here. However, what is occuringfrom both UV and NIR is:ProteinTyrosineFrom NIR comparison of 100.5 and 100.6.There is a shift from the amide/protein nature of 100.5 (titrate decant) to the vinyl region of 1643 in 100.6.Protein/tyrosine being identified via UV at 272 peak absorbance in 100.5.Polymer microspheres 0.25 microns.11/13/23 (100.5 started on 11/08/23)Therefore, 100.5 (decant, titration) is the culture showing the most visible change thus far in the vitals.Appearance of culture has changed from clear to cloudy. pH has steadily decreased. Strong signs of polymerformation under scope.100.5 NIR major feature shows a shift of absorbance from the 1610 nm region to the 1510 nm regeion. It alsolooks like 100.5 shows a relative increase in the 1420 nm region.1610 is the vinyl area1510 is the amide protein area1420 could be in the aromatic region, either ArCH or ArOH.Nov 14 2023It seems that the emphasis is 105.6 is the production either a protein or vinyl or both. It may be that they areone and the same thing.Notes from Lab Log 11/14/23 on the observation of TSP 100.2 after 6 days incubation.-Too early to say the consequences of what is happening here. But we definitely have synth cell productionhere. This means the time to do so has apparently been shortened from 14-20 days to 6 days. 100.2 wasestablished on 11/08/23. We can definitely say the L. enzyme is having an impact upon the culture, whether itwill be for better or worse hopefully we shall learn to the better. What does seem the case is that the cells areof the small variety (~4 microns diameter) and that the color is apparently a grayish color.I have added .08 gms of L.enzyme now to TSP 100.3 to attempt an acceleration of synth cell growth in thatculture also. TSP 100.4 serving as the control shows no such surface growth at this time, so the enzymedefinitely seems to be decreasing the synth cell production time.Alkyl AlcoholProtein Shift from 275nm to 265 nm UVUV Structural ShiftsTDS Shifts under observationNotes from Lab Log 11/14/23 on the observation of TSP 100.2 after 6 days incubation.2023.11.12 Decanted - Vinyl Production?4 -TSP 100.2 Enzyme Addition TSP 100.2Too early to say the consequences of what is happening here. But we definitely have synth cell productionhere. This means the time to do so has apparently been shortened from 14-20 days to 6 days. 100.2 wasestablished on 11/08/23. We can definitely say the L. enzyme is having an impact upon the culture, whether itwill be for better or worse hopefully we shall learn to the better. What does seem the case is that the cells areof the small variety (~4 microns diameter) and that the color is apparently a grayish color.I have added .08 gms of L.enzyme now to TSP 100.3 to attempt an acceleration of that culture also. TSP 100.4serving as the control shows no such surface growth at this time, so the enzyme definitely seems to bedecreasing the production time.TSP 100.6 Titration of TSP 100.0Acidified the precipitate with H2SO4 for UV work.Worked very well.Alkyl alcohol identified.There does seem to be some activity at 1080.This corresponds to an alcohol also, of form RCOH.915 Methyl1643 VinylFrom NIR comparison of 100.5 and 100.6.There is a shift from the amide/protein nature of 100.5 at 1523 nm (titrate decant) to this vinyl region of 1643in 100.6. .2023.11.12 Titration of TSP 100.0Acidified the precipitate with H2SO4 for UV work.Worked very well.Alkyl alcohol identified.There does seem to be some activity at 1080.This corresponds to an alcohol also, of form RCOH.915 Methyl1643 VinylFrom NIR comparison of 100.5 and 100.6.There is a shift from the amide/protein nature of 100.5 at 1523 nm (titrate decant) to this vinyl region of 1643in 100.6. .CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.12 Decanted - Vinyl Production?5 2023.11.14 TSP Culture VariantsI am in process of switching over to a digitial lab notebook as primary. Some paper notes will be carried inparallel but digital lab may easily become primary. Sortable by date, subject, notes, etc. Date entries andsubject entries will both be made.Let's mention some of the complexity that is taking place here. A single pH change introduced into the TSPculture allows for 7 different visual variants to develop, and this is only a start.1. Mature native culture (2 weeks incubation) - top layer where polymer-synth cells (blood) develops2. Mature native culture (2 weeks incubation) - middle layer comprising bulk of solution - to be systematicallyanalyzed with controls. Preliminary work has already been done and is recorded prior to complexity of pH andtemperature control became apparent over a couple of months of work.3. Mature native culture (2 weeks incubation) - bottom layer, white precipitate. Also to be systematicallystudied with.4. Native TSP culture, freshly developed over approx. 3 day period. ph ~3.8.5. Titrated fresh TSP culture to raise pH from native ~3.8 to ~7.23 separates TSP into two components:5-1: Transparent decantable solution of amber color. Forms bulk of solution5-2: Precipitate: greyish in color. Upon centrifuge, it will be found that this grey precipitate actually separatesinto two separate layer: white and grey.5-2-1 Grey precipitate (less dense)5-2-2 White precipitate (more dense)This brings us up to 7 visual variants with a single pH change introduced. No temperature control, roomtemperature on all.Next, we have another variant that develops in the decanted transparent solution, this is number 8:8. In the decanted titrate, after 2-3 days at room temperature, a polymer forms within the solution and turnsthe solution cloudy. All NIR signs are saying that this is a vinyl based polymer. Microscopic examination of thisagrees, as "microspheres" of size approx. 250 nm are now visible under the scope with improved techniques.Vinyl microsphere production of this size is a standard commercial operation.All of the above occurs as a result of a single pH modification with no temperature control. This lets us knowwhat we are up against and why dozens of variants in form are presenting themselves under insufficientlycontrolled and monitored conditions.Let's monitor culture vitals - in spreadsheet.As a prelude to this, here is another observation developing involving 3 culture variations with enzyme study:100.2 This culture applies .04 gm L. enzyme to 150 ml of DILUTE TSP.100.3 This culture applies .04 gm L. enzyme to 150 ml of CONCENTRATED(Evaporated) TSP.100.4 This culture is of CONCENTRATED TSP only.The ratio between the dilute and concentrated TSP is approximately a factor of 3, i.e., the reduction of approx.6000 ml to approx. 2000 ml. This means the impact of the enzyme in the dilute TSP is expected to be greaterthan that of the concentrate TSP. This is where an observation comes in that supports this. TSP 100.2 was tobe discarded in favor of TSP 100.3, but it is found that TSP 100.2 becomes valuable within this observation. Itwill therefore be maintained to some degree.The observation is that there is a color change between the three cultures. 100.2 is lightest in color with awhitish tint, 100.3 is intermediate with a slight white tint, and 100.4 is the darkest solution with an amber tint.I also am seeing some surface development on 100.2. This will have to be examined under the scope. It willpresent a risk to the benefits surmised from the L. enzyme.The signficance of this is that this is one of the many ways it can be determined that the L. enzyme is havingan impact upon the culture. Other methods of determination include:2023.11.14 TSP Culture Variants6 1. "Vitals" monitoring (pH, TDS (total dissolved solids), and PPT (parts per thousand (actually specific gravity))2. UV and NIR monitoring.It is consistently being established that the L. enzyme is having an impact upon the TSP culture from the onset.This is desirable, as interruption of protein development is a primary goal here.Now to vitals in spreadsheet.I have developed a model to establish the "interest level" in a culture. It is too complex to describe here but isbased upon some rather sophisticated probability modeling. Maybe another time, but here are my scores:TSP 100.8 95% DecantTSP 100.9 92% DecantTSP 100.5 62% DecantTSP 100.0 17%TSP 100.4 23%TSP 100.3 12%TSP 100.2 38% Enzyme studyIt took a great deal of work to develop this model but it is worth it to me. All encoded into the culturespreadsheet.This tells me interest level is in the list of100.8 decant100.9 decant100.5 decant100.2 decantIt is very demanding and time consuming to monitor these cultures so it helps tremendously to whittle downthe playing field.One of the first things learned from this is that it will be of higher interest to boost the enzyme concentration in100.3 from ~.04 gms to ~.12 gms to anticipate similar results in the concentrated TSP enzyme trial.Now bear in mind that 100.2 is now showing a significant surface layer development. This could be serioustroubling, don't know, but let's see under the scope.-Too early to say the consequences of what is happening here. But we definitely have synth cell productionhere. This means the time to do so has apparently been shortened from 14-20 days to 6 days. 100.2 wasestablished on 11/08/23. We can definitely say the L. enzyme is having an impact upon the culture, whether itwill be for better or worse hopefully we shall learn to the better. What does seem the case is that the cells areof the small variety (~4 microns diameter) and that the color is apparently a grayish color.I have added .08 gms of L.enzyme now to TSP 100.3 to attempt an acceleration of that culture also. TSP 100.4serving as the control shows no such surface growth at this time, so the enzyme definitely seems to bedecreasing the production time.A question: Is development within TSP 100.5, 100.9 etc vinyl or protein, or BOTH? NIR shows both with sometransitions taking place between the solution form of the titrate and the precipitate form of the titrate. Not sureyet which is dominant in which. Keep reviewing and collecting UV, VIS-NIR and NIR until you sort this out.Uniformity under scope indicated microspheres dominanting at 0.25 spherical diameter, but I think UV and NIRsay both protein and vinyl are active.There are definitely bio vinyl polymers. A hot topic is biodegradable vinyl polymers for use in medicine, i.e., thebody. So we could easily have a combination here.We have:CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.14 TSP Culture Variants7 "The optical absorption measurements for poly(vinyl alcohol) evidenced bands in the UV region (see Figure 2)at 223, 282 and 327 nm assigned to n?p* transition, due to the presence of the C=O group and p?p* transition,respectively which arise from unsaturated bonds, C=O and/or C=C, which are present in the tail-head of thepolymer."https://www.researchgate.net/figure/UV-absorption-spectrum-of-the-polyvinyl-alcohol_fig2_289203733I have a spectrum that shares a great deal in the general sense of what is shown. I am out of range for the 223but it certainly is following the profile. I have absorbance at ~338 and 282 general peak with derivativedetermination.Also we have:"Spectral examination of the solvolysis products of vinyl acetate and 1-methoxyvinyl ester copolymers showsunequivocally that the absorption bands in PVA at 225, 280, and 330 mµ are related to carbonyl-containingstructures."https://onlinelibrary.wiley.com/doi/abs/10.1002/pol.1963.100010412CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.14 TSP Culture Variants8 2023.11.14 TSP culture variationLet's mention some of the complexity that is taking place here. A single pH change introduced into the TSPculture allows for 7 different visual variants to develop, and this is only a start.1. Mature native culture (2 weeks incubation) - top layer where polymer-synth cells (blood) develops2. Mature native culture (2 weeks incubation) - middle layer comprising bulk of solution - to be systematicallyanalyzed with controls. Preliminary work has already been done and is recorded prior to complexity of pH andtemperature control became apparent over a couple of months of work.3. Mature native culture (2 weeks incubation) - bottom layer, white precipitate. Also to be systematicallystudied with.4. Native TSP culture, freshly developed over approx. 3 day period. ph ~3.8.5. Titrated frest TSP culture to raise pH from native ~3.8 to ~7.23 separates TSP into two components:5-1: Transparent decantable solution of amber color. Forms bulk of solution5-2: Precipitate: greyish in color. Upon centrifuge, it will be found that this grey precipitate actually separatesinto two separate layer: white and grey.5-2-1 Grey precipitate (less dense)5-2-2 White precipitate (more dense)This brings us up to 7 visual variants with a single pH change introduced. No temperature control, roomtemperature on all.Next, we have another variant that develops in the decanted transparent solution, this is number 8:8. In the decanted titrate, after 2-3 days at room temperature, a polymer forms within the solution and turnsthe solution cloudy. All NIR signs are saying that this is a vinyl based polymer. Microscopic examination of thisagrees, as "microspheres" of size approx. 250 nm are now visible under the scope with improved techniques.Vinyl microsphere production of this size is a standard commercial operation.All of the above occurs as a result of a single pH modification with no temperature control. This lets us knowwhat we are up against and why dozens of variants in form are presenting themselves under insufficientlycontrolled and monitored conditions.2023.11.14 TSP culture variation9 2023.11.15 TSP Culture VariationsThe cultures are very active and interesting. 100.7 has come to life, this is the petri dish series of the titrationdecant, set up on Nov 12, 72 hrs. The purpose of this series is to determine if surface area exposure is a factoron the polymer formation from the decant. It may note be as much a factor as first surmised as the polymerseems to uniformly distribute throughout the decant over time.One thing interesting is that four cultures, supposedly identical were poured from the same container. Butthere is variability between the four, with one dish showing much higher surface layer development. This is anexample of the high level of variation that can occur with cultures with unknown subtle factors at play.The next observation concerns TSP 100.0, the primary active culture of main volume (~1500 ml). This cultureis now already producing the synth cell-polymer layer in large quantity on the surface. This was abreakthrough development and it has now been replicated on a large scale reliably. It was thought this processrequired a minimum of 2 weeks up to 3 weeks, but it has now taken place in 7 days, culture started on Nov 082023.In addition, the three layer separation is also beginning to form. The three layers are an important feature ofthe mature culture. The three layers are:1. Top: synth cell - polymer layer, whitish in color.2. Middle: bulk of volume, amber color solution.3. Bottom: precipitate layer, whitish to light grey in color.These layer are not yet well defined in TSP 100, but there separation is perceptible. The solution remainsalmost entirely grey. Oxidation will eventually turn the main solution amber.I have my eye on a mid IR instrument, but unfortunately is just not likely to be financially feasible. I have arather extensive library of work available from previous years. If I am unable to acquire (seems likely) there isanother route that might be able to make headway. If I can make tentative identification of a material orfunctional group and can then acquire a sample of that material, then UV and NIR comparison may well besufficient to establish the confirmation level desired.I am already very close to substantiating the claim of polyvinyl alcohol. The factors are:1. UV analysis shows a general match2. MID IR - my plot of "CDB Protein Complex Sep 06 2015" looks to be one remarkable match to that from theSDBS database.3. NIR looks like it is going to match also, but I do not have a reference yet.So to push this confirmation level further:1. Work hard to find a NIR plot of PVA - needs to be 900 - 1700 nm preferably.2. ACQUIRE A SAMPLE of PVA and see if you can get a match. Then run what you have : UV, NIR and see whatyou can do.SOLUBILITY COMMENTS:This version of PVA has a solubility that depends upon pH it would seem. PVA is stated to be water soluble (pHimplied here of ~7). Our proposed PVA is indeed soluble in water. But the synth cell-polymer is not, it is solublein strong acid and possibly acetone, but not in water. These our important characteristics to keep in mind. Alsoour precipitate from the titration may well be a more pure form of PVA. Keep working on it...I found a source for PVA and it should be on the way. Will help a lot. This is a good alternative to access to aMID IR instrument if I can make a good tentative identification. You have never had cultures at this level ofspecificity before.Let's look for PVA reactions in the meantime.There is an interaction between PVA, boric acid and iodine. Working on acquiring PVA and boric acid."Interaction of Poly(viny1 Alcohol), Boric Acid, and Iodine Though it has been known for some time thatpoly(viny1 alcohol)(PVAL), iodine, and boric acid form a c~mplex,'~ no quantitative estimation of thecomposition or the stability values of the complex has been reported. In this communication, we are giving for2023.11.15 TSP Culture Variations10 the first time results regarding the composition, the molar extinction coefficient, and the stability constant ofthe complex. EXPERIMENTAL PVAL solution was prepared by weighing it exactly and dropping it into ameasured quantity of doubly distilled water, raising the temperature to 85OC, and then allowing it to cool.Iodine and boric acid solutions were prepared by standard methods.6 The molecular weight of PVAL was de-termined by a viscosity method.7 The optical density (log Zo/Z) and the spectra were recorded by using aSpecord-UV-VIS spectrophotometer. RESULTS AND DISCUSSION Absorption maxima of the PVAL-iodinecomplex formed in the presence and in the absence of boric acid has been reported'e2 in arbitrary unitsproportional to the absorbance. Zwick3 reported that the absorption maximum is shifted continuously from580 to 700 nm by increasing the con- centration of boric acid at a constant concentration of iodine. But weobserve that at a constant concentration of iodine, the intensity of color increases but the absorptionmaximum remains only at 14,500 cm-' (around 680 nm) and does not change with boric acid concentration(Fig. 1)"https://www.researchgate.net/publication/250427602_Interaction_of_polyvinyl_alcohol_boric_acid_and_iodineGood news, PVA and borax alone have a reaction. This is the "slime" affair.I have produced a reaction with TSP 100.5 and borax. It is creating what appears to be a brittle polymer.Polymer formation by reaction has been achieved.All signs are that this polymer shares some of the fundamental properties of PVA, but what version of polymeris formed is not certain. Brittle nature is important vs a flexible polymer. My take right now might be anacetate form. I have it under the scope photographed at 300x. It appears to have constructed a crystallineform. Application of heat indicates it may melt fairly easily. It appears quite pure.Another important observation. The amount of enzyme added to TSP 100.2 and 100.3 completely changes howthe culture responds. 100.2 has .04gms added to ~ 150ml of TSP 100.0. 100.3 has .12 grams added to ~150ml of TSP 100.0. The results ARE ENTIRELY DIFFERENT. TSP 100.2 now has a full fledged synth cell-polymermatrix that has formed on the top within 24 hrs. TSP 100.3 is now CLARIFIED & TRANSPARENT with no surfacegrowth visible. This indicates the amount of enzyme has made a tremendous difference in the result. A greaterconcentration of enzyme appears to be more destructive to the culture growth process. The difficult factor tounwind is that TSP 100.3 was approx. 3 times more concentrated than 100.1 so I suspected they wouldbalance each other out and act mostly the same. Not so, they have reacted completely differently. One caselooks very unfavorable and the other quite favorable and promising. The control shows the usual ambersolution color.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.15 TSP Culture Variations11 2023.11.16 (Borax Polymer) & 100.6I am looking at 100.8. This is the larger volume decant sample prepared for additional pI testing. It will nowalso be used and given equal priority to polymerized protein production (presumed identity). It is following thesame progression of 100.5 (also a titration decant) with:1. Increasing cloudiness2. Lowering of pH3. Increased TDS4. Increased PPTas recorded on spreadsheet during vitals monitor today.We notice that the "vitals" in no way capture the whole story. They are only a helpful indicator but they canmiss very important events. An example is what is happening between 100.2, 100.3 and 100.4100.4 is TSP control100.2 is .04 gms enzyme in dilute TSP100.3 is .12 gms enzyme in conc. TSPThe vitals indicate possible significance with TSP 100.2 which is true and of interest since it is forming a synthcell-polymer surface layer even though lighter concentration of enzyme has been added.However, 100.3 by vitals shows nothing of particular interest. Visually, however, it is more than important,especially as it compares to 100.2 The only difference should be an increased concentration of enzyme to 0.12gms. The culture is TRANSPARENT with NO synth cell - polymer surface layer. This is a huge difference, but thevitals miss it and only observation detects it. So stay alert is the lesson....Now let's review the NIR spectrum of the borax-decant reaction from yesterday. It was presumed that it mighthave close ties with a polyvinyl compound. We did notice and are aware, however, that the borax reactionproduct was neither water soluble or flexible.Note: (1) Weak peak (3) Strong peakTabular info comes from CRC NIR handbookPeak info:983 (2) OH from water (maybe but maybe not....) (979)Phenolic OH (990)1170 (2) Alkene, polyenes (1170)1347 (2) Methyl (1360) larger delta than normal1436 (3) Aromatic amine (1432)Methylene (1440)1480 (3) Polyamide (1480)Secondary Amine (1481)Amide/Protein (1483)and in general, high absorbance in the range of 1436 - 1480; aromatic amine is strong here.Now my initial thought was that polyvinyl alcohol related compounds might show up in this NIR spectrum.However, the OH is not dominant here if at all, and the compound is not soluble. The water question can besettled and separated out by sustained drying of the sample. Phenols are moderately soluble in water. Now weknow that this compound is highly insoluble. This indicates that OH solubility is not a strong factor here. Thepolymer and protein signatures are what dominate here. For now, I will presume water over phenol in the 983nm group but will keep an eye on this.Most conductive polymers are polyenes. Polyenes are a main target of astrobiology. Polyenes share somechemical reaction properties with PVC vs PVA. PVC is C2H3Cl (notice no OH in any way). So if of a plastic2023.11.16 (Borax Polymer) & 100.612 nature would seem to be more likely of PVC family than PVA since we are insoluble and somewhat crystallinenature under scope and brittle.THE MATERIAL DOES NOT PASS A CONDUCTIVE TEST WITH THE OHMMETER AT THIS POINT.so 983 peak remains undecided between OH water and alkene, polyene.Now the aromatic amines, polyamides, secondary amine, and amide protein all speak of being a polymerizedprotein, especially in light of the borax reaction which has produced it.This exists as the most current interpretation of this material that has been created, a polymerized protein.Unclear at this point if the polymerization is likely to come from aromatic or amide origin, or both for thatmatter. We know that in the end a "rubberized" polymer is developed, but this can come from the other TSPsurface synth-cell polymer layer. But out of curiosity, is rubber aromatic or amide based?Rubber is alkene based. OK, this gets very interesting. Synthetic rubber is a copolymer between styrene andbutadiene. Styrene is a VINYL group attached to an aromatic ring. Butadiene is an alkene.Guess what, all the signs are starting to point the same direction. Aromatic, vinyl, alkenes are the componentsof synthetic rubber, which is most characteristic of the mature clot gross properties. Aromatic, vinyl, alkenesappear to be dominant structures of our NIR plots. Do not keep trying to force a single functional group. Thecombination of these three functional groups along with synthetic blood go a long way toward explaining theculture findings in general.Start looking at these three functional groups in a combined sense, understanding that separations in theculture will likely break these into parts. We will now keep an eye on the NIR plots more closely with thesevarious combinations.Let's create more of the proposed polymerized protein, now designated as TSP 100.10. Done. Drying.I am trying to acidify TSP 100.10 material w/ moderately strong H2SO4. It is difficult. It may be slightly solublein H2SO4 but certainly not easily. This means that both polymeric forms, TSP surface layer synth cell-polymerlayer and this newly developed polymer from the decant and borax are BOTH very much insoluble in water anddifficult to solubilize in reagents as well. Just like the clot was....I need to conduct solubility tests on 100.10 just like I did on the synth surface polymer on TSP 070. More studythere again as TSP 100.0 matures.My goal is to attempt to dissolve (hydrolize) TSP 100.10 for a UV run to determine if protein is detected.OK, that was straightforward. Definite absorbance at 275 nm. Protein confirmed. Polymeric protein remainsconfirmed. Now our goal is to try and determine the nature of the polymer. Our first NIR run says we have:1. alkenes, polyenes2. methyl possible3. aromatic amines4. methylene5. polyamide5. secondary amine6. amide/protein7. polyamide, amideTherefore, the reduced and presumed polymer set is:1. polyenes2. aromatics3. amines4. polyamideSeems like an ideal polymerized protein to me. Qualifies as a constituent of the final clotting process. Missingquality seems to be the rubber nature, but this seems to fit quite well for the TSP 070 surface synth layer -polymer.The essence of the clot seems to be established at this point:1. Synth erythrocytes w/hemoglobin2. Two polymeric forms, one for structual density and the other for elasticity. Both highly insoluble, even inmany strong reagents/acids/caustics.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.16 (Borax Polymer) & 100.613 This is a major achievement. It would certainly be of benefit to understand how the two polymers may jointogether.Now the rubber property would be imparted by the vinyl group, which we now understand is a part of thecopolymeric process of synthetic rubber. We may well have, and are expected to have a copolymerarrangement here.This means that we would expect to see the vinyl component from one of the two polymers, and the strongestcandidate for that is now the TSP synth cell - polymer surface layer. Let's look at NIR plots.Guess what? (The database sure pays off). From notes of Nov 12, the vinyl component is in the precipitate!This is the other half of the TSP prior to titration!. We have all the parts we need. We have therefore separatedbetween:Three polymers, consistent of all needed parts:One separated polymer contains:1. polyenes2. aromatics3. amines4. polyamide(i.e., proteins and binding polymers)The second polymer, i.e. the precipitate of the titration:1. contains vinyl(i.e., imparts elasticity)and synthetic blood containing hemoglobin in a polymer matrix, also elastic in nature (thus protein & polymerhere also).This means you need to know if the precipitate contains protein also. I think NIR says yes. But let's work onthat one.This is all leading to the likely conclusion of three polymeric protein complexes:1. Decant2. Precipitate3. Surface synth cell layerEach one will have its own dominant chemical properties.This would therefore certainly be a copolymeric operation. Expected that decant + preciptitate = synth cellsurface layer with synthetic blood.The precipitate to the titration is 100.6. We have enough of this material to work with for now. It is importantto realize that this layer, upon centrifuge, actually contains two layers, an upper grey layer higher in volume,and a lower white layer minimal in volume. This layer now becomes important in seeking out the vinylcontribution to the overall polymer structure. For now we will focus only on the grey upper layer as itcomprises the bulk of the volume of centrifugation.We have it. NIR evaluation of the borax polymer vs the precipitated tells us the distribution and make up of thesynth surface layer.TSP 100.6 (the precipitate from titration) has high absorbance in the 1640 region. This is the vinyl dominantregion of NIR.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.16 (Borax Polymer) & 100.614 TSP 100.10 (the borax polymer) has high absorbance in the 1440 region. This is dominated by aromatic amine.The precipitate is not high in protein content. 293 nm and 232 peaks were identified in the 100.6 acidifiedprecipitate.So what this is saying is that the middle layer of the culture (most volume) contributes the bulk of protein. Theprecipitate contributes the elasticity.So our three main components (everything is temperature and pH dependent for expression (~18 expressionsthus far) are:1. Synthetic blood (w/hemoglobin) (scores of proteins within blood) (surface layer of mature culture)2. A set of proteins that can be polymerized with borax, structurally firm but not elastic. (within decant)3. A vInyl contribution for elasticity of the culture. (within precipitate)This makes up the majority of the clot structure.1,3-Butadiene (a synthetic rubber) from NIST data base UV spectral search shows a peak at 293. This is a vinylcompound and then a match for our spectrum of TSP 100.6 at this point (not the rest of the spectrum nor our232 peak found.NIST and Chemspider both have some UV searching capability if you can get a part of the name. VERYVALUABLE!NOTE:The styrene UV spectrum explains high absorbance towards the 232 peak[NOTE DEC 08 2023 I SEE NO BASIS FOR THIS PREVIOUS STATEMENT - IT IS NOT ACCEPTED AT THIS TIME].Synthetic rubber is a copolymer between styrene and butadiene.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.16 (Borax Polymer) & 100.615 2023.11.17 Solubility MethylationThe picture looks increasingly clear. It would seem that we have a genetically engineered co-polymerizationprocess taking place here. The process involves synthetic blood, an aromatic protein polymer, and a syntheticrubber equivalent polymer. Their origin is the Cross Domain Bacteria (CDB). The result is increased coagulationor a clotting of the blood. The end result is definitely a lethal threat.Need to outline for panel and conditions of disclosure.Lab needs:1.Look at TSP 070 on surface layer polymer. Expected now to be a combination of aromatic protein polymerand synthetic rubber (vinyl).2. Solubility studies on aromatic protein polymer. Can iron be used to polymerize something like Boron (Borax)did?Boron is a metalloid with 3 valence electrons."There are many forms of polymerization and different systems exist to categorize them.""In more straightforward polymerizations, alkenes form polymers through relatively simple radical reactions;""Step-growth and chain-growth are the main classes of polymerization reaction mechanisms. The former isoften easier to implement but requires precise control of stoichiometry. The latter more reliably affords highmolecular-weight polymers, but only applies to certain monomers.""The manner in which polymerization is conducted is a highly evolved technology. Methods include emulsionpolymerization, solution polymerization, suspension polymerization, and precipitation polymerization."3. Methylation - put an end to a polymer chain reaction, it would seem? Can you induce methylation in the lab?Mitigation implications. Ultrasound now understood more clearly also. NAC, Vit C, Citrate influence on culturesagain.4. Consider setting up the next paper: (?)Cross Domain Bacteria (CDB) : Synthetic Blood, BIoplastics, and the Global Blood Clot5. Head toward replication of TSP 070 activity, namely nucleic acids and bacteriophage.6. Increased understanding of enzyme influence. Concentration appears dramatic. Work towards enzymeisolation.7. Begin exploitation of NIST UV Spectra, can be very helpful. Suggest modeling of synthetic rubber bycombining UV plots of styrene and butadiene.UV Spectra on NIST site can be extremely helpful. Have not been aware of them. Some level of compensationfor lack of MID IR instrument exists with this information. If you get to the point of prospective chemical group,e.g., vinyl, NIST can be used to bring up related UV spectra. This can add an important layer of confirmation toNIR work (and generalities) by bringing a chemical group into the picture of confirmation instead of onlyfunctional groups. No knowledge yet of affordable NIR data base for structure but functional group analysis isin good position with CRC handbook.Looking at the comparison NIR plot between human blood and synthetic blood is most intriguing. There ismuch to be studied here. An acceptable reference spectrum for blood is now in question. However, the synthblood spectrum is not in doubt as to what it contains. Dramatic differences exist, and they are, in part at least,now anticipated and predictable.2023.11.17 Solubility Methylation16 Notice how a major feature of the decant (e.g. 100.5 and 100.9) is that they gradually transform from clearamber toward white as a polymerization takes place. This dynamic change will be important to get a handleon. Vitals and pI (variable value?) are two important tools as well as UV monitoring. Is the protein polymerchanging structure here and can you show this? It is almost certain to be the case.The refractometer will hold its zero calibration value even after turning off. This is great, saves a lot of extratrouble. With respect to vitals, refraction is ending up being the most sensitive indicator with a variation of~20% within cultures over time. TDS is next with a variation of ~8%. pH is the least senstive, with a variationof ~ 3%. All are useful, especially when combined. The idea of adopting an RMS as a reference frame for mydeveloped methods in probability analysis seems to be very convenient and logical and simple to implement.For the record of years past, the final work from work years ago resulted in the probability model of:Pr% = (200/pi) * atan(c * delta x)andc = 1/(Pr range) * tan(Pr level% * (pi/200))VALUES IN RADIANS.In this most recent implementation of my model, the RMS of the data set is used as the Pr range value. The Prlevel adopted for that reference range is 71%. Peak Probability will be 1.414 * Pr%. This is a logical referenceframe to adopt as an alternative to E90 or z=1 score, etc. This original work was done years ago and hasshown itself to be a valuable and useful model to many applications. Much simpler to implement than thestandard distribution curve and it is far more realistic to accomodate extreme values or handle the "long tail"problem.Culture change can be modeled very well with this model. Changes in any parameter can be modeled in adifference from a reference culture value. Differences can then be converted to a daily slope. Daily slopes canthen be modeled from a probabilistic sense using the model above and the interest level of each culture canbe established for each sample. Overall interest level can be assessed with the average of the three differentvitals monitored.This model can be applied rather easily to any situation of interest, e.g, financial, data samples, etc, any timeseries data.In terms of application of this model, TSP 100.5 and 100.9 are indeed high interest cultures. There is a lot ofchange going on with respect to the vitals as polymerization occurs. This change process can undoubtedly bestudied (and learned from) in great detail. It is an area where interference of protein development can beanticpated with sufficient knowledge of internal biochemistry taking place.TSP 100.5 is now pure white, compared to original amber color. Time span 7 days. TSP 100.9 is now a paleamber, time period 5 days.The paper has been started.Human Transformation : Synthetic Blood, Bioplastics,and the Global Blood Clot Clifford E CarnicomNov 17 2023CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.17 Solubility Methylation17 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.17 Solubility Methylation18 2023.11.18 Borax polymer solubilityA few solubility tests have been run on the borax polymer today. Results are:1. in strong acid (H2SO4), not soluble2. in strong alkaline (NaOH-KOH), slowly soluble3. in acetone, not soluble4. in ethanol, not solubleThese are helpful results. It has not been published yet, however, the deceased blood clot was eventuallydissolved in NaOH-KOH solution over a period of ~ 2 months. This was recorded on Aug 11 and Aug 13 2023 inVol 30 of the laboratory notebooks.This now becomes important as we have similar solubity behavior between the deceased blood clot (the mostdense and developed, rubber polymer properties) and the borax polymer that has been developed. We knowthat KOH-NaOH has generally been our only successful reagent for dissolution of any of the filaments. Weknow they help to break down the disulfide bonds, as in hair.A question is what other types of polymer bonds might they break down? In this case borate bonds areinvolved. So we have a good question coming up with the polyvinyl-borax polymer that can be made (the"slime" case). Will KOH-NOH work on that as well? If so, it shows it is effective on the polyvinyl bonds andborax, not just disulfide.Yes, lye does break down slime. YT video demo:https://www.youtube.com/watch?v=abJ1ETlhZ-wHe needed add some water instead of so much direct powder lye, but it still demonstrates the point. Waterwas used at the end and more clearly shows that breakdown has occurred.This is valuable as we are not dealing with disulfide bonds here. Vinyl bonds are involved, as they are in ourcase also.This was a valuable trial here. Another linkage in bond disruption between the clot polymer and one of theculture polymer forms.Back to our questions of yesterday then:Lab needs:1.Look at TSP 070 on surface layer polymer. Expected now to be a combination of aromatic protein polymerand synthetic rubber (vinyl).2. DONE Solubility studies on aromatic protein polymer. Can iron be used to polymerize something like Boron(Borax) did?Boron is a metalloid with 3 valence electrons.DONE. SEE LAB NOTES ABOVE.3. DONE Methylation - put an end to a polymer chain reaction, it would seem? Can you induce methylation inthe lab? Mitigation implications. Ultrasound now understood more clearly also. NAC, Vit C, Citrate influence oncultures again.MUCH BETTER UNDERSTANDING IN PLACE. METHYLATION IS ACROSS THE BOARD IN THE HUMAN BODY.AFFECTS MOST EVERYTHING. ANY DISTURBANCE IN METHYLATION IS GOING TO CAUSE MAJOR PROBLEMS.4. DONE Consider setting up the next paper: (?)Cross Domain Bacteria (CDB) : Synthetic Blood, BIoplastics, and the Global Blood ClotTitle changed to:Human Transformation: Synthetic Blood, BIoplastics, and the Global Blood ClotPAPER HAS BEEN STARTED.5. Head toward replication of TSP 070 activity, namely nucleic acids and bacteriophage.6. Increased understanding of enzyme influence. Concentration appears dramatic. Work towards enzymeisolation.2023.11.18 Borax polymer solubility19 7. Begin exploitation of NIST UV Spectra, can be very helpful. Suggest modeling of synthetic rubber bycombining UV plots of styrene and butadiene.8. DONE Blood Evaluation. Results highly favorable.I want to start learning about methylation. Here we go, it is bigtime right away:"DNA methylation is a biological process by which methyl groups are added to the DNA molecule. Methylationcan change the activity of a DNA segment without changing the sequence. When located in a gene promoter,DNA methylation typically acts to repress gene transcription""As of 2016, two nucleobases have been found on which natural, enzymatic DNA methylation takes place:adenine and cytosine."It is exactly what I envisioned. It does not apply only to an alkyl chain by any means. it is taking place here onan cyclic rings and aromatic rings as well. This says that it is quite feasible that methylation increase in thebody may well assist in the disruption of an aromatic polymer protein. This is very favorable information. Andby the way, DNA is a polymer as well. Yes, it absolutely is, it is a polynucleotide. Photos below.Basically methylation helps to regulate DNA to stay healthy and thwart mutations."Epigenetics, the study of the chemical modification of specific genes or gene-associated proteins of anorganism.""The principal type of epigenetic modification that is understood is methylation (addition of a methyl group).Methylation can be transient and can change rapidly during the life span of a cell or organism"A gene is a segment of DNA that contains sequences of many bases, varying in size from a few hundred to 2million. Each gene affects a specific aspect of health. For example, some genes contain instructions on how tomake specific proteins.A gene is a specific segment of DNA that tells cells how to functionDNA is the genetic material or code that tells cells in the body how to replicate themselves. Almost every cellin an organism!s body contains a complete copy of its genome, packaged into chromosomesMethylation is a massively important topic in human health. There is a lot of ground to cover here:"What is methylation?At the most basic level, methylation and demethylation refer to the transfer of methyl groups#simplestructures of one carbon and three hydrogen molecules (CH3)#to and from various bioactive compounds in thebody. These compounds (e.g., proteins, enzymes, hormones, and more) must be methylated in order tofunction optimally or to create other substances required by the body.For example, methylation is essential for the production of certain bioactive vitamins (e.g., folate and vitaminB12), amino acids, neurotransmitters, hormones, red blood cells, DNA, RNA, and antioxidants that directlyimpact cardiovascular, neurological, reproductive health, energy production, detoxification pathways, andmore.""Methylation influences just about every essential process in the body. It's needed for the production of aminoacids, neurotransmitters, hormones, red blood cells, DNA, RNA, and antioxidants; for proper detoxification ofhormones and toxins; and for determining whether certain genes will be expressed or not (the power ofepigenetics). It's a big deal.But when this vital process is compromised (which can happen as a result of a variety of dietary, lifestyle, andgenetic factors) your physical and mental well-being, from a cellular to whole-body level, will pay the price.The good news: A simple homocysteine test can help you get a glimpse of your own methylation cycle healthand help determine whether you need to make any dietary or lifestyle changes#"https://www.mindbodygreen.com/articles/what-is-methylationCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.18 Borax polymer solubility20 "What does methylation do to DNA, and why does it mater? As a metabolic process, it basically switches geneson and off and repairs DNA.This is a big deal, because gene expression has the power to influence many aspects of health, including one!srisk for certain diseases, such as neurological issues and some cancers.Methylation and demethylation, which is the opposite process, basically work together to control differentfunctions throughout the body."https://draxe.com/health/methylation/So quite easy to see how methylation is on par with enzymes in mitigation strategies.Let's look at our blood to keep pulse on status.Report: Blood status looks highly favorable. Methods make sense, pH regulation, methylation, proteininterruption, electrolytes.,salicylateWorking on research paper. Most dramatic blood culture images (the original appearance) is recorded on Sep19 2023. Actually occurred in TSP 01. Phenomenal result.I have a very significant plot now to look at. It is a NIR comparison of TSP 100.5 (Decant) over a 12 dayincubation period. This is from the time that it is decanted (amber color) until it turns white approx 10 dayslater.Main feature I am looking at is that appears there is a bulk of mass (absorbance) shift from the Nov 06 TSP100.5 (amber) at a very broad peak of ~1620 +/- 20 nm to a more focused peak at Nov 18 TSP 100.5 (White)of 1516. They do however, continue to share a great deal, and in fact the majority of the turf from 1530 - 1650nm.Interpretation of this is:1620 is alkene and vinyl1520 is protein, 1515 is polyamidethis makes a lot of sense. It is saying it starts out stronger in the vinyl section but then shifts over to a proteinnature. However, it still retains the vinyl nature. This means that a conglomerate is being formed, i.e. apolymerized protein as your surmise.This is exactly what you envision happening. This answers a very important question on culture dynamics.this makes a lot of sense. It is saying it starts out stronger in the vinyl section but then shifts over to a proteinnature. However, it still retains the vinyl nature. This means that a conglomerate is being formed, i.e. apolymerized protein as your surmise.This is exactly what you envision happening. This answers a very important question on culture dynamics.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.18 Borax polymer solubility21 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.18 Borax polymer solubility22 2023.11.19 Project PlanningLots going on, as usual.The paper is posted, I regard it as a good mark on grounding. Steady she goes..Adjust the list.1. Done: solubility study on borax polymer. All perfectly consistent w/ KOH-NaOH impact.2. Done: Methylation is at the heart of health. On solid ground3. Done : Paper: Human Transformation: Synthetic Blood, BIoplastics, and the Global Blood Clot10. DONE PDF of 2023 papers revised. Review lab notebook publishing schedule4. UV Spectra on NIST are a new asset. Can we find NIR database?5. Head toward replication of TSP 070 activity, namely nucleic acids and bacteriophage.6. Enzyme concentration studies. Enzyme isolation studies.7. MIxing the culture into blood in various ways. Impact?8. Keep sorting out the chemistry of the culture dynamics and progression1. Native---pI series investigation?2. Decant1. pI series investigation?2. Decant progression- transformpI series investigation3. . Precipitate4. Surface layer - synth cells - polymer5. NIR of Human Blood vs Synth layer - sort this out - alcohol?9. Duplicate TSP 0701. Nucleic acids2. Bacteriophage.The general sequence coming up is:1. Nucleic acids, bacterophage2. Health Lineage3. Panel on:1. Culture2. Mitigation4. Laboratory notes up - phasedQuite the software battle today. Need a database that will kick out a report that autosizes for printing a2023.11.19 Project Planning23 variable text field, such as this one. No dice, not an easy one. Jasper report may be a candidate but nobody upfront doing this on regular basis. Very surprising. You have a problem on this. Database lab log is perfect formy use, but not for distribution. Access and other heavy duty dababase progams will do it, but not the simplesoftware. Crystal reports, JasperReport. Astounding how difficutl that is.OK fantastic. I have found it. Online converter. Amazing service, files look great.Use CS View if need to whittle file, and then the online converter, or maybe just straight with the whole file.Beautiful solutionl.https://cloudconvert.com/Cloudconvert.com is my ticket. I have created an account. I am free to write uninterrupted now.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.19 Project Planning24 2023.11.20 UV NIR TSP 70 Nucleic Acid4. DONE UV Spectra on NIST are a new asset. Can we find NIR database? - NO.There simply are none that are available affordably. $1500 min for 1000 spectra, always with cost. NIR istaking off in popularity due to its ease of use. But you are on your own with respect to spectral interpretation.5. Head toward replication of TSP 070 activity, namely nucleic acids and bacteriophage.6. Enzyme concentration studies. Enzyme isolation studies.7. MIxing the culture into blood in various ways. Impact?8. Keep sorting out the chemistry of the culture dynamics and progression1. Native---pI series investigation?2. Decant1. pI series investigation?2. Decant progression- transformpI series investigation3. . Precipitate4. Surface layer - synth cells - polymer5. NIR of Human Blood vs Synth layer - sort this out - alcohol?9. Duplicate TSP 0701. Nucleic acids2. Bacteriophage.10. Construct a UV & NIR plot of polyvinyl alcohol?The general sequence coming up is:1. Nucleic acids, bacterophage2. Health Lineage3. Panel on:1. Culture2. MitigationThis is the topic to work on today:1. Native---pI series investigation?I will compare TSP 070 (Mature native culture Oct 19) against TSP 100 (Native Nov 08). Am interested in all 3issues: pI behavior, UV and NIR.First observation is that TSP 070 is transparent, TSP 100 is cloudy. This makes sense in that we know a matureculture eventually separates into 3 layers: top polymer, middle bulk of volume, bottom precipitate). Titrationand decanting is an entirely separate process of separation. For now only time and gravity are involved in thisanalysis.UV: On Nov 05 2023 we collected a UV spectrum on TSP 070, the mature culture. We had an absorbance of259 nm. We looked up on this and we found a strong existence of the literature on nucleic acid absorbance at259 nm. We have it again, as solid as can be withTSP 070, absorbance well defined at 259 nm.OK, there are VERY important differences between the UV spectra of TSP 070 (Mature) and TSP 100 (underincubation, no complete separation into 3 layers yet). TSP 070 (Mature) has a very clearly defined peak at 2592023.11.20 UV NIR TSP 70 Nucleic Acid25 nm. TSP 100 does not have this YET and instead has peaks at 273 and a very broad shallow peak at ~315. Soquite different, and the absorption at 273 once again indicates likely tyrosine presence. This is therefore a veryimportant difference to notice and to monitor. 315 nm is a little hard to pin down since it is no broad andshallow. Let's see if we can find a UV spectrum of DNA.This information leads us to suggest that the direction of the mature culture will be the creation on nucleicacids. Next step is to monitor the second culture, i.e., TSP 100 and seed if the event repeats itself. Also, anyother contenders for 259 nm absorption? What does NIR say? What is the UV profile of DNA? What is the NIRprofile of DNA?Ref UV DNA easy to come by:Images attached.https://www.researchgate.net/figure/UV-absorption-spectra-of-native-DNA-black-and-glycated-DNA-incubated-for-1-day-with-01_fig1_281243983https://www.researchgate.net/figure/UV-absorption-spectra-of-native-DNA-DNA-lysine-photoadduct-and_fig1_40728359I would say the case is pretty strong.Also found NIR of DNA:https://www.sshade.eu/data/SPECTRUM_OP_20181101_05should be looking for a NIR peak of about 1400 nm.possible a valley also near 1100.if you can come up with a good idea of what something is, you can often find a reference spectra to determineif you are right or not. This method is acutally preferable to having a full IR spectrum and NOT having any ideawhat it is. This shows the value of being able to come up with a good estimate of what might be involved. Casefor nucleic acids is developing because of this method, although originally you had no idea that DNA might beinvolved or even detectable with the culture. It may be.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.20 UV NIR TSP 70 Nucleic Acid26 2023.11.21 Nucleic Acids (?) - DNA- PVAThere is a good problem before us. Are we dealing with Nucleic Acids/DNA in TSP 70 or are there alternativessuch as alcohols or aromatics involved? Why does the human blood NIR have such a broad strong peak in theapparent alcolhol section? If you do have DNA/nucleic acids would you be able to isoate it with chilled alcohol?Is there anything else know to absorb so strongly at 259 nm? How do you reconcile the NIR plot of TSP 70 withUV plot and qualitative chemistry? How do you interpret the dynamics of culture change, as revealed by NIRcomparison, between TSP 70 (mature) and TSP 100 (developing)? From analysis of the reference NIR plot, weexpect to see a peak at ~1455. We have a broad shallow peak in that region but the peaks at 1500 and 1600swamp 1455 out. Need to discern between all three.A polyvinyl NIR andUV plot is now going to help you sort out the aromatic - alcohol problem.We also have the NIR culture dynamics of TSP 070 vs TSP 100 now available. The shifts between 1600 and1500 are very important now to identify as to what is actually changing. Alcohol - Aromatics - Vinyl?Would TSP 100 mature more quickly if incubated? Room temps are pretty low now at night. Notice, however,that TSP 100 is indeed showing the changes of maturation. Color change and 3 layer separation is now readilyvisible. Also in the spare TSP 100 material. We definitely have solid replication of maturation; process takes ~2 weeks to become definite and identifiable.Our previous list :1. Head toward replication of TSP 070 activity, namely nucleic acids and bacteriophage.2. Enzyme concentration studies. Enzyme isolation studies.3. MIxing the culture into blood in various ways. Impact?4. Keep sorting out the chemistry of the culture dynamics and progression1. Native---pI series investigation?2. Decant1. pI series investigation?2. Decant progression- transformpI series investigation5. . Precipitate6. Surface layer - synth cells - polymer7. NIR of Human Blood vs Synth layer - sort this out - alcohol?8. Duplicate TSP 0701. Nucleic acids2. Bacteriophage9. Construct a UV & NIR plot of polyvinyl alcohol?The general sequence coming up is:1. Nucleic acids, bacterophage2. Health Lineage3. Panel on:1. Culture2. MitigationA very smart move to acquire Polyvinyl Alcohol. Strong easy spectrum to acquire, 4% solution. 500ul in 3mlmore than sufficient for strong UV spectrum. Peaks at 240 and 268. Distinctive spectrum, not like anythingseen thus far. THe 240 peak shifted quite a bit with a strong concentration, the 268 held steady. The shape ofthe profile is another distinguishing feature to pay attention to in UV we see. If you were to just take the peaknumber of 268 you might want to say protein. VERY FAR FROM THE TRUTH, very far away from protein. Yousee one of your strong clues is the profile or slope of the spectrum, nothing like protein that we have seen. Sonow you pay close attention to this and it is all the more reason you must do your best to identify a referencecandidate and FIND THE UV SPECTRUM for that candidate before you draw strong conclusion based upon peakinfo alone, and often singular peak info.2023.11.21 Nucleic Acids (?) - DNA- PVA27 Now let's compare acquired PVA spectrum with what someone else said was a reference.By direct measurement and acquistion of 99.9% PVA I have the following:230 (220, 230)very strong 269 (270) weak 277 (280) weak (now revised with extended quartz cuvette use).Now I notice I have another small peak at ~ 277 nm. With the slopes of my curve considered, this is also amassive departure even from the claimed 282. SO THIS IS A RADICAL DIFFERENCE. Let's look at NIST. NISTdoes not have UV. Another name for polyvinyl alcohol is Ethenol (not Ethanol!).and guess what, it does not match at all with what is being claimed as a reference. This is important anddamages confidence in "reference data".This is the reference polyvinyl alcohol (PVA) information I have located before. This took place in Nov 14 notes(first day of digital lab notes):We have:"The optical absorption measurements for poly(vinyl alcohol) evidenced bands in the UV region (see Figure 2)at 223, 282 and 327 nm assigned to n?p* transition, due to the presence of the C=O group and p?p* transition,respectively which arise from unsaturated bonds, C=O and/or C=C, which are present in the tail-head of thepolymer."https://www.researchgate.net/figure/UV-absorption-spectrum-of-the-polyvinyl-alcohol_fig2_289203733I have a spectrum that shares a great deal in the general sense of what is shown. I am out of range for the 223but it certainly is following the profile. I have absorbance at ~338 and 282 general peak with derivativedetermination.Also we have:"Spectral examination of the solvolysis products of vinyl acetate and 1-methoxyvinyl ester copolymers showsunequivocally that the absorption bands in PVA at 225, 280, and 330 mµ are related to carbonyl-containingstructures."https://onlinelibrary.wiley.com/doi/abs/10.1002/pol.1963.100010412Neither NIST or Chemspider has a reference UV spectrum for polyvinyl alcohol. That is a surprise. NIST has MidIR. Another thing we learn is that Chemspider requires the chemical name to find something, it does not likepopular names. The chemical name for polyvinyl alcohol is ethenol which I had no idea of.So we have a little bit of an issue. I have UV by direct measurement. Reference claims of absorption are notworking well at all. Right now I trust my direct measurement the most.Another source is giving of PVA dissolved in water (just like I am doing) with peaks of:231, 281, and 324.https://link.springer.com/article/10.1007/s40089-015-0150-yanother source gave (see above);223, 282 and 327 nmanother source gave:225, 280, and 330 mµThe average reference from all three of these sources is: 226, 281, 327.Now I can not measure 226 easily so that one is out.Let's redo my spectrum from 250 to 350. The 324 peak is shown to be weak. The 281 is much stronger. Let'ssee if we can isolate these. Is ther any possibility I have contamination? It would not seem likely to me.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.21 Nucleic Acids (?) - DNA- PVA28 I have run it again. I have well defined peaks at: 268.5 276.5.These simply do not match the reference claimed. The 276.5 is distinctly separate, with respect to slope, of theclaimed 281 nm. The 327 does not even show up. This is entirely perplexing. I have a purchased commercialproduct that is stated to be, acts like, looks like, makes a film like PVA. But my own spectrum acquired directlywith no apparent difficulties does not match stated reference spectra. I am glad that I am doing this but find itto be MOST puzzling.OK, we are going to run another reference spectrum of some sort with a different material. There is simply noreason to doubt my instrument at this point.Isopropyl alcohol should be an example, but NIST and Chemspider does not have this either.I found NIR of isopropyl alcohol but not UV at:https://www.researchgate.net/figure/Isopropyl-alcohol-spectrum-generated-in-10-milliseconds-and-are-compared-to-Pacific_fig7_252229308OK, I am going to use acetic acid, vinegar, as a reference control. In addition I actually have 2 quartz cuvettesthat wil extend my range down to 190 nm. This is a crucial advantage. I will buy two more, take advantage ofthem, and I will extend my UV range considerably.For special and important cases, I will now be able to use quartz cuvettes to get down to 190 nm.This is great. A 25% increase in capability of the instrument for $25.Acetic acid comes out somewhat OK. I get a strong peak at 200 nm, NIST shows it at 210, another shows it at212, but at least we are in the neighborhood and can get significantly extended range now. Let's go back toPVA with the quartz cuvette. If you are willing to use 1 cuvette at a time your 4 cuvettes should last indefinitelyif you take care of them.OK, this change was incredibly valuable to make. I have not matched PVA where it counts. I have the peak nowat 230 nm. Close enough and shows me the kind of error that can exist even under very good conditions. Whathappens here is the peak at 327 simply does not exist for me. The peak at 281 is very weak, exists but poor tobank on. I have an additional peak at 269 that no one is talking about, also weak but more more defined.However, at 230, it is off the chart, and absorbance is of 2 magnitude. Extremely strong with 1 drop PVA in 3ml cuvette. You would simply never see this without the extended range, and the magnitude of this peak is somuch greater than the others that this peak alone is what serves as the reference match point. And now wehave matched PVA.This brings in a new lease to UV analysis. I may discover quite a bit more now with UV analysis of most anysample. Lot's that can go on below that 235 nm point.... This really does change things.Now we can move on. We now have uniqueness of PVA available with combination of UV and NIR card hasbeen made (nice film appearance).I think we need to remeasure UV on TSP 70 and TSP 100 with extended cuvette range and then combine withthe NIR data to sort out whether we have alcohols or aromatics involved.A DNA spectrum:https://www.degruyter.com/document/doi/10.1515/nanoph-2020-0225/html\Measured UV:PVA : 230 (220, 230)very strong 269 (270) weak 277 (280) weakNow for the chilled alcohol DNA test:I see absolutely no sign of DNA strands being formed and rising within chilled alcohol being placed on the topof TSP 070.There is, however, a definite precipitate being formed so an important reaction of some type is taking place.Recall our early test with alcohol that also produced a precipitate. Never investigated further. The nature ofthis precipitate will need to be determined. The precipitate forms in the TSP 070 portion of the culture-alcoholCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.21 Nucleic Acids (?) - DNA- PVA29 mixture, not in the alcohol. Further indication that it does not involve DNA, but a protein or polymer is verylikely. The ethanol may therefore not need to be chilled at all but it remains so for storage right now until testis made with room temperature ethanol.The precipitate does centrifuge to the bottom of the tube within the TSP 070 solution. Alcohol stays clearabove. Let' start by looking under the scope.There is very little material to work with, just enough to get under the scope.And yet another form. My vote is protein. It is composed of very small needle crystals, about .4 um in thicknessand 6-10 um long. Interspersed with occasional groups of undersized synth cells.I would say the Nucleic acid - DNA hypothesis is going out the door and that we likely have a protein insolution. Not sure what the ~210 nm UV absorbance is referring to yet. But combined with NIR work, this isgoing the direction of an aromatic protein within TSP 070, not an alcohol.[NOTE FOR DEC 09 2023: A THOROUGHLY CHILLED ALCOHOL DNA TEST GIVES A POSTIVE RESULT. NOSETTLING PRECIPITATE FORMED].[ALL 3 TESTS FOR DNA ARE POSTIVE: UV, NIR (HYDROXYL GROUP), CHILLED ALCOHOL)CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.21 Nucleic Acids (?) - DNA- PVA30 2023.11.22 Solubility-Health-Blood(Syn)I am seeing that the CRC NIR reference is not as specific as it appears to be. Acquiring a direct measurementcontrol NIR plot of Polyvinyl Alcohol (PVA) is turning out to be very insightful. CRC Handbook cannot be countedon as the almighty NIR reference that I had presumed it was. This introduces a major level of caution as well asadditional verification required on any NIR claims that are made. I will demonstrate this point with an analysisof the pure, directly acquired and measured PVA. CRC will be a very useful reference, but not an absolutereference, at it will now be combined more readily with the generalized charts such as from Galaxy Scientific.(The image icons within this database serve as major prompts; they must be opened jointly and separatelywithin an image editor or viewer)TO DEMONSTRATE THE WEAKNESSES IN THE PROCESS:Now let's make a standard analysis with the info I have available from below and measured directly:UV :PVA : 230 (220, 230)very strong 269 (270) weak 277 (280) weak TSP 070 213 261269 280 peaks indicate protein230 somewhat unknown but as we recall it is a good indicator of protein folding changesso these are both in support of protein.NIR:Peak 1201 (1200) 1489 (1490) 1574 (1570) 1669 (1670) Broad Absorption Peak 1450-1600Valley: 1005 (1000, 10010) 1284 (1280)Valley at 1005 would indicate LACK of secondary alcohol (-CH-OH) (1004)LACK of primary aromatic amine (1003)LACK of OH from tertiary alcohol (-C-OH) (1006)Peak of 1201 indicates OH from water (1200) (sample is dried thoroughly)Methyl (1195, 1194)Valley at 1284 would indicate LACK of [NO CRC SIGNAL LISTED 2012]Peak at 1489 indicates primary aromatic amine (1489.5)primary aromatic amine (1487)secondary amine (1486)amide/protein (1490)OH polymeric (1490)amide/protein 1492aromatic amine (1493) (1493.5)Peak at 1574 indicates polyamide (1570)amide/protein (1570)alcohol or water (1580)alkyl alcohol or water (1583)Now what we see here is the preponderance of information points VERY strongly toward the existence ofprotein.However, here is the chemical formula for polyvinyl alcohol:C2H4Oor[-CH2CHOH-]nand the Lewis diagram is insightful as it shows how the chain forms with alternating H, OH bonds.NOW THE BIG LESSON HERE IS THAT THERE IS NO PROTEIN WHATSOEVER INVOLVED. A PROTEIN ASSESSMENTWOULD BE ENTIRELY ERRONEOUS EVEN THOUGH THERE IS SUBSTANTIAL INFORMATION TO ACCEPT THATINTERPRETATION.So our job is to study this situation more thoroughly to prevent this from ever happening.1. FIrst lesson is no matter how many entries there are for a certain category that does not necessarily mean2023.11.22 Solubility-Health-Blood(Syn)31 that is what it is at all. A single alternate entry can (and is here) the proper choice (in this case "polymericalcohol" at 1490. Truth is the correct choice was spot on, you just needed to be observant to catch it.Therefore everything you know, see and can learn is important in the process, and in this case, physicalobservation and behavior and solubility of the material is key information in the process.2. Second lesson is that UV was a total loss and detour on both accounts. Protein inference was incorrect onBOTH accounts, folding and blip in the 270-280 region. I would say the shape of the UV profile is now asimportant as the peak absorbance. Certain classes of materials will likely exhibit similarity in slope profiles.NIST data base might be useful here but you can see how much even reference spectra can vary dependingupon scales used. This also shows the value of the colorimetric route on proteins - this should settle thequestion.What this means is that you need to be very CAREFUL at all times about being duped into something. You mustcheck, check, cross check and put all your wits together, and independent methods and controls are ALWAYSessential. No guessing or surmising is allowed when you draw the final conclusions.Now if you play your cards right, NIR actually gives us a perfect answer:1. Methyl group (does seem likely or necessary at the end of the chain.2. polymeric alcohol3. alcoholIf you play it wrong, you call it a protein because of redundancy and inference from reference sources.andMID IR WOULD HAVE LIKELY EASILY SETTLED THE AFFAIR BUT YOU DO NOT HAVE IT NOW.Now the lesson from all this is to back up and review what you know and what might be assumptions or biasinfluence.Back to our list.9. DONE Construct a UV & NIR plot of polyvinyl alcohol? (and we learned a great deal from it, didn't we...?)1. Head toward replication of TSP 070 activity, namely nucleic acids (now very questionable) andbacteriophage (still seems likely).2. Enzyme concentration studies. Enzyme isolation studies.3. MIxing the culture into blood in various ways. Impact?4. Keep sorting out the chemistry of the culture dynamics and progression1. Native---pI series investigation?2. Decant1. pI series investigation?2. Decant progression- transformpI series investigation5. . Precipitate6. Surface layer - synth cells - polymer7. NIR of Human Blood vs Synth layer - sort this out - alcohol?8. Duplicate TSP 0701. Nucleic acids2. BacteriophageThe general sequence coming up is:1. Nucleic acids, bacterophage2. Health Lineage3. Panel on:1. Culture2. MitigationThree good problems:1. Try now to settle out the nucleic acid -protein issue. 190nm should also help.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.22 Solubility-Health-Blood(Syn)32 2. NIR human blood -synth later comparison (sort out confusion there)3. Bacteriophage propositionLet's revisit this human blood - synthetic blood issue.Microscope settings that seem improved or best at 3200x.1. Only refers to screen center2. Condenser set at about 1/2 of eyepiece, eg. 40x=.65 set at .323. Light diaphram set in lower range eg 2-74. Exposure about 1/45. Gain looks to be close to 0.6. White balance about 1/67. Tint about 40%8. Hue 459. Saturation 3010. Brightness 9011. Contrast 9512 Gamma 100Fine adjustment of exposure with diaphragm.I did try, for the first time, to withdraw fluid from the sac of fluid that builds up in my lower feet. Very shallowneedle entry, at most 1/8 inch. Was not able to withdraw any fluid. Small drop of blood appeared on surfaceafter withdrawal. Looked under scope at blood, highly variable results, slide preparation not standard so willhold off judgement. Definite CDB presence, however.In weeks past, I was able to evaluate a MID IR blood sample of mine I had taken in Nov of 2015. As I recall, Iwas not able to identify any significant abberation in the functional groups expected between 3826 and 1536cm-1 (disregarding 1536-600 fingerprint region). It would certainly be of value to have MID IR again but that isalso quite a luxury financially. A true reference for blood has been lost by now. The fingerprint region would bea very important part of the picture to analyze. This comparison with human blood reference spectra wasmade on Oct 31 2023 in handwritten notebook Vol 33. I actually have close to 100 pages of handwritten notesin addition to these digital notes that were started on Oct 30 2023. So there is much lab work that is notwritten until the major switchover on Nov 16 2023. Also a comparison was made in hand notes on Nov 032023.I have been trying to relocate my notes about the solubility tests on the polymer-synth layer. They took placeon Nov 04 2023 hand notes. OK, resutl was that they polymer-synth layer is soluble in strong H2SO4, NOTKOH-NaOH. Also in acetone to some degree. Projection was made that this will be useful for LC work later on.Now here is where the conflict is arising. You are saying that you identified that the blood clot dissolves overtime in KOH-NaOH. You are also saying apparently that the blood clot did not dissolve in H2SO4.But now on Nov 04 2023 you are saying explicitly that the polymer-synth layer only dissolved in strong H2SO4and possible acetone, but definitely did not and does not dissolve in KOH-NaOH.THIS IS A HUGE DIFFERENCE AND A BREAK ON LOGIC. The presumption is that the polymer-synth layer and theblood clot would be subject to a similar breakdown mechanism, i.e., both dissolve in lye or both dissolve inacid. These notes say this is not the case.THIS IS A BIG FLAG AND THIS NEEDS TO BE EXAMINED PRONTO...Here are some other notes from Nov 18 (digital notebook) that are relevant:"These are helpful results. It has not been published yet, however, the deceased blood clot was eventuallydissolved in NaOH-KOH solution over a period of ~ 2 months. This was recorded on Aug 11 and Aug 13 2023 inVol 30 of the laboratory notebooks.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.22 Solubility-Health-Blood(Syn)33 This now becomes important as we have similar solubity behavior between the deceased blood clot (the mostdense and developed, rubber polymer properties) and the borax polymer that has been developed. We knowthat KOH-NaOH has generally been our only successful reagent for dissolution of any of the filaments. Weknow they help to break down the disulfide bonds, as in hair."So look at the above and what it is saying. It is saying the polymer-synth layer dissolves only in H2SO4 directly(and maybe acetone) and NOT base, but then when the borax polymer is formed IT dissolves in base. This isgetting very curious. The borax polymer is formed by combination of borax and TSP 100.5 (hand notes on Nov15 2023). The polymer formed is called 100.10. 100.5 is the decant, NOT the polyjmer-synth layer."These are helpful results. It has not been published yet, however, the deceased blood clot was eventuallydissolved in NaOH-KOH solution over a period of ~ 2 months. This was recorded on Aug 11 and Aug 13 2023 inVol 30 of the laboratory notebooks.This now becomes important as we have similar solubity behavior between the deceased blood clot (the mostdense and developed, rubber polymer properties) and the borax polymer that has been developed. We knowthat KOH-NaOH has generally been our only successful reagent for dissolution of any of the filaments. Weknow they help to break down the disulfide bonds, as in hair."So look at the above and what it is saying. It is saying the polymer-synth layer dissolves only in H2SO4 directly(and maybe acetone) and NOT base, but then when the borax polymer is formed IT dissolves in base. This isgetting very curious. The borax polymer is formed by combination of borax and TSP 100.5 (hand notes on Nov15 2023). The polymer formed is called 100.10. 100.5 is the decant, NOT the polyjmer-synth layer.OK we have 2-3 different compounds going on here reacting with acid and base. Let' sort these out:100.5 is the decant.100.10 is the borax polymer (made with borax combined with 100.5)The clot dissolves in KOH-NaOH slowly.The borax polymer (using 100.5 and borax) disslolves in KOH-NaOH.The polymer-synth layer dissolves in Strong H2SO4 (and maybe acetone) but NOT in KOH-NaOH.Also I am recalling the decant forms a precipitate when combined with a carbonate. See those questioningnotes also - what if they do not dissolve in the body as well?This says that the polymer-synth layer and the borax polymer are two very entirely different things. The boraxpolymer is apparently behaving closer to the clot in terms of acid-base behavior vs the borax polymer and thepolymer-synth layer.Think about this: We know that the decant mixed with borax creates a polymer. This polymer is very hard todissolve, requiring lye.Now you also tried an experiment with carbonate and the decant. You used washing soda (sodium carbonate).Apparently you wanted to see what would happen if it was placed in a base solution? It would be great to recallCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.22 Solubility-Health-Blood(Syn)34 what you motive was that started that process. The point here is that it created a precipitate. So the questionis how soluble is that precipitate? How do you know it does not build up in the body as well? What is thatprecipitate made of, other than to presume it is a carbonate form. Well, what are kidney stones? I suspectcalcium carbonate....There are potentially some serious issues with any use of carbonates, such as baking soda, washing soda.These are both carbonates, sodium as well. Could easily bring too much sodium into the body. It would only beable to be used sparingly. Think about this:Decant + carbonates are producing a precipitate. (baking soda)Decant + borax is producing a polymer.Neither one of these is desired in the body. Watch out - health risks here. Presents a dilemma when trying toincrease alkalinity in the body.Excess of baking soda can also cause edema, salt imbalance here."In many species the pancreatic duct epithelium secretes HCO3- ions at a concentration of around 140 mM bya mechanism that is only partially understood."You can work with this concentration amount and equate it to baking soda ingestion.Let's look at our NIR plot we made comparing synth layer to human blood:Human blood has (all statements will be relative in this study) high absorbance in the 900-950 nm regionrelative to polymer-synth layer. Synth refers to synthetic blood detected within the polymer layer. Synth layeris white and insoluble except in lye and possible partial in acetone.CRC 900-950 nm is dominated by the methy groups and methylene. I presume this is the fatty acids in blood.On Oct 31 and on Nov 03 2023 I compared human blood Mid IR data with reference data. I had a match with allgroups on my Mid IR spectra with the reference blood on the upper portion of the spectrum. Lipids, or alkanesare a part of that. So I do not see a problem here with normal blood absorbing in this region. The NIR spectraof the polymer-synth layer, however, shows very low relative absorbance here. This is our first majordifference.The next place we show NIR absorbance in blood is at 1524 nm, and it is relatively low. The 70% level of thepeak spans from about 1500-1550nm. Our closest match here is 1520 which has amide by CRC and this alsowould be a match to human blood.This is all we get out of the human blood NIR spectrum and it remains a match with expectations, as the MID IRplot of 2015 does. So no real surprises here.However, when we look at out polymer-synth layer we have an entirely different story. It has a very strongpeak at 1520 which matches both amide and POLY amide. But very high relative to human blood. In additionwe have a very strong broad peak centered about 1604. This is in the polyamide vinyl region (1598-1613) CRC.Also at 1700 we have higher methyl absorbance (even though we cannot see peak) than the polymer synthlayer. This is also consistent with the 900-950 nm region findings.Conclusion: for polymer-synth layer relative to human bloodLikely to have methyl disruption (i.e., decrease in fatty acids?)Likely to polymerization of amides taking placeLikely to have vinyl groups activeFatty acids have many health benefits, especially w.r.t. cardiovascular disease, cell membrane health, etc.Excess baking soda probably not a good idea, fish oils may be. VIt B complex, enzyme research, etc. Acidityproblems remain.Would be helpful to look for methyl disruption in human samples.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.22 Solubility-Health-Blood(Syn)35 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.22 Solubility-Health-Blood(Syn)36 2023.11.23 Blood(Syn)-Polymer CompareForefront topics are:1. Clot solubility conflict : polymer synth layer vs clot solubility2. Comparison between human blood and synth layer3. Bacteriophage prospectThese are major topics.Lots of issues coming up as usual.Carbonate precipitation with decant is now raising some important questions. Carbonate ions may be aproblem, e.g, baking soda. Also excess sodium with baking soda another problem. Also the false boraxpromotion issue. Look like significant health issues with any precipitate or polymer that forms. How would youknow they would dissolve - polymers not, and carbonates only in acids in general. You need to find out thenature of the precipitate with sodium carbonate added to the decant.Is there a magnesium carbonate? Yes, there most certainly is. Can cause diarreaha but could be a candidate.Mg citrate would also double up on the magnesium ion, so need to watch that on intestinal fortitude...Magnesium carbonate has poor absorption, it is stated. Too much magnesium in the blood dangerous.Need to run a culture control with magnesium citrate alone.Right dosage is likely important.Edema taking place in right foot on afflicted ankle/leg at night. Could be salt imbalance in connection withaffliction.Alka selter tablet has about 1 gm citric acid, 2 gms baking soda [~2.5 mm in vial) (and aspirin)Mg citrate has 100 mg Mgpink saltAM urine pH 5.3- 5.6. Too low. Want to get this up to 6.5 or so with minimum addition. Also increase waterintake.Body temp running avg. 97.5, not too bad. 96.5 is. 98 better.EnzymesVit B, CHuman blood-synth layer comparison to continue.Mid IR exploration of vinyl group with historical "CDB Protein Complex" IR data (cerca 2015) looks to be mostimportant. Initial peak match search brings up following candidates as:1. Butyl vinyl ether C6H12O2. Cyclohexyl vinyl ether C6H14OThese candidates have essentially exact matches with IR collected Sep 2015 of "CDB Protein Complex".Out of about 10 candidates from entire SDBS database. These are lowest molecular weight. I am guessing wehave something here.Look at physical properties, synthetic rubber, etc.Combination of human-synth blood comparison with polymerization probable is a worth topic.Let's get back to the blood comparison.Recalling from yesterday:Conclusion: for polymer-synth layer relative to human blood:1. Likely to have methyl disruption (i.e., decrease in fatty acids?)2. Likely to polymerization of amides taking place3. Likely to have vinyl groups active2023.11.23 Blood(Syn)-Polymer Compare37 andWould be helpful to look for methyl disruption in human samples.and always allow for error and mistakes!Main mitigation strategies are:1. Interfere with protein development2. Interfere with polymerization3. Interfere with acidity3. Support methylation4. Disrupt polymer structure within and below skin4. and the usual of everything before, antioxidation, etc.5. and now added on Dec 05 2023 the new information coming to light on acidic amino acids and pI resarch onblood proteins.We also learned some very important acid-base properties and that things are behaving quite differently:OK we have 2-3 different compounds going on here reacting with acid and base. Let' sort these out:100.5 is the decant.100.10 is the borax polymer (made with borax combined with 100.5)The clot dissolves in KOH-NaOH slowly.The borax polymer (using 100.5 and borax) dissolves in KOH-NaOH.The polymer-synth layer dissolves in Strong H2SO4 (and maybe acetone) but NOT in KOH-NaOH.This says that the polymer-synth layer and the borax polymer are two very entirely different things. The boraxpolymer is apparently behaving closer to the clot in terms of acid-base behavior vs the borax polymer and thepolymer-synth layer.Now let's look at UV for both human blood and the synth layer. Also look at the synth layer again under thescope.The UV spectrum of blood is far more interesting than I would ever have imagined. It is the most active UVspectrum I have encountered thus far. Busy all the way across from 190 nm to 400 nm. Here are peaks:Human Blood : 232 (230) 275 (270, 280) 346 (350) ~400+(another peak developing)We know 230 is commonly associated with protein peak folding. 275 would be classic protein area. 346unknown now.And here is interesting info, but it does not match our finding. We have a strong peak at 345, 346, not 365."Journal of Forensic SciencesJ Forensic Sci. 2020 Sep; 65(5): 1716#1721.Published online 2020 Apr 28. doi: 10.1111/1556-4029.14439Ultraviolet 365 as an Alternative Light Source for Detection of Blood SerumThese results show that ultraviolet (UV) at 365 nm (UV365) was effective in the detection of even smallamounts of blood plasma and serum, compared with UV at 395 nm, which was not. UV365 was also found tobe useful in distinguishing blood imprints from clotting blood which had been transferred to material versusblood that had been added directly. Taken together, these results demonstrate that UV365 may be utilized asa simple, nondestructive method for blood serum detection."Now the difference alone is what of greatest interest to me. I have a very clear strong peak at 345 nm, not365. 365 is closer to the bottom of a valley for me. So why this difference? What is 345 measuring and what is365 measuring? I could not imagine using 365 right now compared to what I am seeing at 345.I have found a good lead on interpretation of UV of blood (in this case it is mouse blood):"UV-Visible absorption spectra of blood and EB diluted in water. A. Five microliters of a BALB/c mouse bloodCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.23 Blood(Syn)-Polymer Compare38 were drawn and diluted to 250 m L (1:50 dilution) with distilled water, centrifuged and the supernatantcollected for analysis. Indicated peaks are: 1) protein amide backbone and nucleic acids (220 nm); 2) proteinswith chromophoric amino acids and other small chromophoric molecules (280 nm); 3) globin-heme interaction(340 nm); 4) soret band (420 nm); 5) oxyhemoglobin b -band ( , 540 nm); and 6) oxyhemoglobin a -band ( ,575 nm). Arrows show the common wavelength to measure hemoglobin (540 nm) and EB (620 nm).doi:10.1371/journal.pone.0110551.g001https://www.researchgate.net/figure/UV-Visible-absorption-spectra-of-blood-and-EB-diluted-in-water-A-Five-microliters-of-a_fig1_267273300So we have the following:peaks are:1) protein amide backbone and nucleic acids (220 nm);2) proteins with chromophoric amino acids and other small chromophoric molecules (280 nm);3) globin-heme interaction (340 nm);4) soret band (420 nm)and ours measured:Human Blood : 232 (230) 275 (270, 280) 346 (350) ~400+(another peak developing)So we have:1. A very general match at 220-230 nm region (also note nucleic acid in addition to protein reference).(remember mouse blood)2. A decent acceptable match at 275 - 280 nm for proteins, although some shift (remember reference here ismouse blood)3. A decent acceptable match at 340 - 345 nm. (remember mouse)4. Notice our external match quite likely developing at ~400 nm.So this is quite decent and gives us some acceptable UV references for blood.Now let's start looking at the synth layer.I have some very good photos of the synth cells under the scope. It takes some doing. First step is that slideneeds to be mildly flushed with water to clean up the surrounding polymer solution. Then the best results willbe obtained as the slide dries from the water under the cover slip. Most cells will congregate and not makegood viewing however some stray cells will provide a good view.The microscope at 8000x is working best at:1. Height of stage q tip height with one end cut off. This step is also critical. 1/8 inch change seems to affectgreatly.2. Brightness of lower dial lamp will have moderate impact, usually between 5-10.3. The diaphragm is set at 0.3 (1/4 of what the objective says)4. Exposure Timed 40%5. Gain 20%6. Color temp 20%7. Tint 45%8. Hue 45%9. Saturation 30%10. Brightness 95%11. Contrast 100%12. Gamma 100%I also have a good macro picture of the polymer synth layer on a glass slide. So good, we have1. UV of human blood, very distinctive.2. Microphotos of synth blood. Easy to compare to human blood photos. 8000x.3. Macro photo of synth polymer layer.4. We have a NIR comparison that demonstrates the polyamide/vinyl nature of the synth blood. Likelymethylation disturbance at play.5. Mid IR of vinyl groups is coming into play6. Macro photo or culture dish surface layer.7. UV comparison spectra between human blood and synthetic blood, high correspondence.8. Photo of "blood red" color change upon mixing polymer surface layer and strong H2SO4CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.23 Blood(Syn)-Polymer Compare39 also recall kastle meyer, hemoglobin, and pI determination.A couple more phenomenall things have happened.1. When you take the dried synth material and put in in strong H2S04, it turns immediately deep red. Slightpurple tint. Hard to avoid the situation with blood. I have photo.2. In addition, there is significant overlap in the UV spectrum with human blood, especially from 190 nm to 300nm. Peaks, valleys and general profile coincides as well.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.23 Blood(Syn)-Polymer Compare40 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.23 Blood(Syn)-Polymer Compare41 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.23 Blood(Syn)-Polymer Compare42 2023.11.24 Abstracts AI - NIR SynthI have done a good job today of learning to prepare abstracts for all papers of 2023. ChatGBT is integratedwithn a newer version of my word processor. I have given it a run, and it works quite well. BIggest difficultywas getting a suitable format for intergration within Wordpress and Substack, not the generation of theabstracts.Process is:1. An Abstract database has been made. Can now be organized by title, date, link, abstract. Very helpful.2. Output will come from the database itself. Convert file to HTML (great option available) and make sure it willpresent on a portrait view ok.3. This html can be loaded with Firefox from the hard drive. Firefox seems to be superior to Brave in severalrespects.4. It can then be uploaded into both Wordpress and Substack and works quite well on both.5. Last step is to attach the links recorded in the database to the titles of the papers.Steps of generating abstract are generally:1. Copy the paper into the Starmaker wordprocess (new and latest greatest version with ChatGBT integrated.2. Choose the summarize text option, mid length.3. Copy the summary into the database, with any minor changes required.A really helpful and valuable addition. Still took lots of work but should be even a little smoother next time.Suggest we work on one year at a time.OK, we are continuing to look at a comparison between the human blood and synth blood.1. I am curious, how much H2SO4 must be used to generate the red color?2. Why did this material not evaporate or dry on the ceramic plate, even after overnight? It has unusualvolatility properties.3. We have NIR plots of the polymer vs the acidified red version of the polymer - synth layer.It is very difficult to produce the purple - red color from the polymer. The color is actually a rich deep purple. Itrequires extremely concentrated sulfuric acid to produce it. Not sure yet if the heat generated in theacid-water reaction is a part of or requirement in the process. It requires about 1 ml of full strength H2SO4 withapprox. 1 ml of water. I am not sure if drying the polymer-synth layer assists the process or not, I have addedboth dried and fresh polymer. It is a remarkable color.So two properties are now unusual:1. Deep rich purple, or purple red color in concentrated sulfuric acid.2.The material seems very difficult to dry and/or vaporize.The polymer does seem to have a very stout vinyl type quality as it is dried."In most cases, purple blood is caused by a rare condition called sulfhemoglobinemia, where sulfur atoms bindto the iron in hemoglobin. This prevents hemoglobin from carrying oxygen effectively, causing a purplish hueto the blood.""Bacterial InfectionsCertain bacterial infections, like endocarditis or sepsis, can also turn blood purple. Bacteria producesulfur-containing compounds that bind to hemoglobin, inducing sulfhemoglobinemia.""Sulfhemoglobinemia is a condition in which sulfhemoglobin, a molecule formed when sulfur binds tohemoglobin, is present in the blood. This molecule gives the blood a purple color. Normally, hemoglobin carriesoxygen throughout the body, and it has a red color when oxygenated and a darker red color whendeoxygenated.However, when sulfhemoglobin forms, it changes the conformation of hemoglobin, altering its ability to bindand release oxygen. This leads to a decrease in oxygen-carrying capacity and a change in the color of theblood. The purple color is a result of the interaction between the sulfur and the iron in the hemoglobinmolecule."H2SO4 added to human blood turns it brown, which is expected. It is oxidizing the iron.However, H2SO4 added to the polymer turns it purple to purple red.2023.11.24 Abstracts AI - NIR Synth43 "When sulfuric acid is added to blood, the resulting color change can vary depending on several factors. Thecolor change is primarily due to the reaction between the acid and the components present in the blood.If the blood contains a high concentration of hemoglobin, which is responsible for carrying oxygen in red bloodcells, the reaction with sulfuric acid can result in a purple color. This occurs because the acid causes theoxidation of hemoglobin, leading to the formation of a purple-colored compound called hematin.On the other hand, if the blood has a lower concentration of hemoglobin or if other substances are present,such as bilirubin or other breakdown products of red blood cells, the reaction with sulfuric acid can result in abrown color. This is because the acid can react with these substances, leading to the formation ofbrown-colored compounds.It's important to note that the color change observed when sulfuric acid is added to blood may also beinfluenced by the concentration and purity of the acid used, as well as the specific characteristics of the bloodsample. Therefore, variations in the color change can occur in different situations.""When blood is exposed to sulfuric acid, it undergoes a chemical reaction known as a"deoxyhemoglobin-sulfuric acid reaction." This reaction causes the blood to turn purple.Sulfuric acid is a strong acid that can denature proteins, including hemoglobin, which is responsible forcarrying oxygen in our blood. Hemoglobin normally carries oxygen in its oxygenated form, which gives bloodits bright red color. However, when sulfuric acid comes into contact with hemoglobin, it breaks down thestructure of hemoglobin and converts it into a different form called deoxyhemoglobin. Deoxyhemoglobin has adarker color, appearing more purple or maroon.""Artificial blood, also known as synthetic blood or blood substitute, refers to a substance that can temporarilyfulfill the functions of natural blood in the human body. It is primarily developed to address the shortage ofdonated blood and to provide an alternative in emergency situations.There are two main types of artificial blood: oxygen-carrying solutions and oxygen-carrying particles.1. Oxygen-carrying solutions: These solutions are designed to carry and deliver oxygen to tissues, similar tothe red blood cells in natural blood. They are typically composed of hemoglobin-based molecules orperfluorocarbons. Hemoglobin-based solutions use modified hemoglobin molecules derived from human oranimal sources, while perfluorocarbons are synthetic compounds that can dissolve and carry oxygen.2. Oxygen-carrying particles: These particles are microscopic in size and are engineered to transport oxygen.They can be made from various materials, such as lipids, polymers, or nanoparticles. Oxygen-carrying particlesare designed to mimic the oxygen-carrying capacity of red blood cells.Artificial blood has several potential advantages over donated blood. It can be produced in large quantities,ensuring a consistent supply. It can also be stored for longer periods and does not require blood typing orcross-matching, reducing the risk of transfusion reactions. Additionally, artificial blood is free from infectiousagents that can be transmitted through donated blood.However, the development and approval of artificial blood have faced numerous challenges. One major hurdleis replicating the complex functions of natural blood, such as regulating blood pressure, clotting, and immuneresponses. Ensuring the safety and efficacy of artificial blood has also been a significant concern.While there have been advancements in the field of artificial blood, it is important to note that it is not yetwidely available for clinical use. Extensive research and rigorous testing are still ongoing to overcome theexisting limitations and bring artificial blood to the forefront of medical practice.""When it comes to blood, bacteria and synthetic biology can also play a role. For instance, bacteria can beengineered to produce specific proteins or enzymes that could be used in blood-related applications such asclotting factors or blood substitutes. Synthetic biology techniques can also be employed to modify blood cellsor create artificial blood components for various medical purposes."An interesting one:"The Carnicom Institute is a non-profit research organization that focuses on the study of geoengineering,aerosol operations, and related health issues. It was founded by Clifford E. Carnicom in 2008 and is based inNew Mexico, United States. The institute conducts independent scientific research and provides educationalresources to raise awareness about the potential risks associated with geoengineering activities.Cross-domain bacteria, also known as "Morgellons," is a term used to describe a condition where individualsexperience unusual skin sensations and the presence of fibers or filaments embedded in their skin. Someindividuals believe that these fibers are a result of a new type of bacteria that can infect both humans and theCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.24 Abstracts AI - NIR Synth44 environment. However, it is important to note that the existence and nature of cross-domain bacteria are still asubject of debate and controversy within the scientific community. Further research is needed to fullyunderstand this phenomenon."NIR of synth polymer layer:1196 Methyl (1194, 1195) weaker peak"Methyl groups typically exhibit absorption peaks in the near infrared (NIR) region around 1196 nm."1521 Amide/Protein (1520) Secondary Amine (1520) Polyamide (1515)"In the near infrared region, the amide bonds exhibit specific absorption bands around 1521 nm. By measuringthe intensity of the absorption at this wavelength, NIRS can provide information about the presence andconcentration of proteins in a sample."1581 Alcohol (1580) alkyl alcohol (1583)"The wavelength you mentioned, 1581 nm, falls within the near-infrared range and is commonly used in NIRSfor alcohol analysis. At this wavelength, alcohol molecules absorb light in a characteristic manner, allowing forthe quantification of alcohol concentration."1609 Vinyl (1613) Polyamide (1618)"In the case of vinyl, NIRS can be used to study its absorption properties in the near infrared range, specificallyat a wavelength of 1609 nm"".Overall, the use of NIRS at 1609 nm can help in characterizing vinyl and understanding its absorption behaviorin the near infrared range."1580-1609 Broad PeakThis is saying that we have a protein/polyamide vinyl, alkyl alcohol dominanting the composition. This would allbe consistent with solubility and physical properties. We seem to be on track quite well here.Now for the polymer synth layer in STRONG H2SO4 (Red-purple color) non volatile -sulfhemoglobinemia(hemoglobin is a part of the reaction).1440 Methylene (1440) aliphatic hydrocarbons, OH Aromatic (1420), OH Phenolic (1420)Work on this with physical propeties, UVCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.24 Abstracts AI - NIR Synth45 2023.11.25 Human Synth Blood CompareI am working on developing a comparison between human blood and the synth blood-polymer layer. Let'srecap the factors that have surfaced:FROM PREVIOUS DAYS:From NIR Analysis: Conclusion: for polymer-synth layer relative to human blood:1. Likely to have methyl disruption (i.e., decrease in fatty acids?)2. Likely to polymerization of amides taking place3. Likely to have vinyl groups active"UV-Visible absorption spectra of blood and EB diluted in water. A. Five microliters of a BALB/c mouse bloodwere drawn and diluted to 250 m L (1:50 dilution) with distilled water, centrifuged and the supernatantcollected for analysis. Indicated peaks are: 1) protein amide backbone and nucleic acids (220 nm); 2) proteinswith chromophoric amino acids and other small chromophoric molecules (280 nm); 3) globin-heme interaction(340 nm); 4) soret band (420 nm); 5) oxyhemoglobin b -band ( , 540 nm); and 6) oxyhemoglobin a -band ( ,575 nm). Arrows show the common wavelength to measure hemoglobin (540 nm) and EB (620 nm).doi:10.1371/journal.pone.0110551.g001https://www.researchgate.net/figure/UV-Visible-absorption-spectra-of-blood-and-EB-diluted-in-water-A-Five-microliters-of-a_fig1_267273300So we have the following:peaks are:1) protein amide backbone and nucleic acids (220 nm);2) proteins with chromophoric amino acids and other small chromophoric molecules (280 nm);3) globin-heme interaction (340 nm);4) soret band (420 nm)and ours measured:Human Blood : 232 (230) 275 (270, 280) 346 (350) ~400+(another peak developing)So we have:1. A very general match at 220-230 nm region (also note nucleic acid in addition to protein reference).(remember mouse blood)2. A decent acceptable match at 275 - 280 nm for proteins, although some shift (remember reference here ismouse blood)3. A decent acceptable match at 340 - 345 nm. (remember mouse)So this is quite decent and gives us some acceptable UV references for blood.Now let's start looking at the synth layer.I have some very good photos of the synth cells under the scope.I also have a good macro picture of the polymer synth layer on a glass slide. So good, we have1. UV of human blood, very distinctive.2. Microphotos of synth blood. Easy to compare to human blood photos. 8000x.3. Macro photo of synth polymer layer.4. We have a NIR comparison that demonstrates the polyamide/vinyl nature of the synth blood. Likelymethylation disturbance at play.5. Mid IR of vinyl groups is coming into play6. Macro photo or culture dish surface layer.7. UV comparison spectra between human blood and synthetic blood, high correspondence.8. Photo of "blood red" color change upon mixing polymer surface layer and strong H2SO4also recall kastly meyer, hemoglobin, and pI determination.9. MID IR correlation with non fitting albumin peaks.2023.11.25 Human Synth Blood Compare46 A couple more phenomenall things have happened.1. When you take the dried synth material and put in in strong H2S04, it turns immediately deep red. Slightpurple tint. Hard to avoid the situation with blood. I have photo.2. In addition, there is significant overlap in the UV spectrum with human blood, especially from 190 nm to 300nm. Peaks, valleys and general profile coincides as well.It is very difficult to produce the purple - red color from the polymer. The color is actually a rich deep purple. Itrequires extremely concentrated sulfuric acid to produce it.So two properties are now unusual:1. Deep rich purple, or purple red color in concentrated sulfuric acid.2.The material seems very difficult to dry and/or vaporize.The polymer does seem to have a very stout vinyl type quality as it is dried."In most cases, purple blood is caused by a rare condition called sulfhemoglobinemia, where sulfur atoms bindto the iron in hemoglobin. This prevents hemoglobin from carrying oxygen effectively, causing a purplish hueto the blood.""Bacterial InfectionsCertain bacterial infections, like endocarditis or sepsis, can also turn blood purple. Bacteria producesulfur-containing compounds that bind to hemoglobin, inducing sulfhemoglobinemia.""When blood is exposed to sulfuric acid, it undergoes a chemical reaction known as a"deoxyhemoglobin-sulfuric acid reaction." This reaction causes the blood to turn purple.Sulfuric acid is a strong acid that can denature proteins, including hemoglobin, which is responsible forcarrying oxygen in our blood. Hemoglobin normally carries oxygen in its oxygenated form, which gives bloodits bright red color. However, when sulfuric acid comes into contact with hemoglobin, it breaks down thestructure of hemoglobin and converts it into a different form called deoxyhemoglobin. Deoxyhemoglobin has adarker color, appearing more purple or maroon."NIR of synth polymer layer:1196 Methyl (1194, 1195) weaker peak"Methyl groups typically exhibit absorption peaks in the near infrared (NIR) region around 1196 nm."1521 Amide/Protein (1520) Secondary Amine (1520) Polyamide (1515)"In the near infrared region, the amide bonds exhibit specific absorption bands around 1521 nm. By measuringthe intensity of the absorption at this wavelength, NIRS can provide information about the presence andconcentration of proteins in a sample."1581 Alcohol (1580) alkyl alcohol (1583)"The wavelength you mentioned, 1581 nm, falls within the near-infrared range and is commonly used in NIRSfor alcohol analysis. At this wavelength, alcohol molecules absorb light in a characteristic manner, allowing forthe quantification of alcohol concentration."1609 Vinyl (1613) Polyamide (1618)"In the case of vinyl, NIRS can be used to study its absorption properties in the near infrared range, specificallyat a wavelength of 1609 nm"".Overall, the use of NIRS at 1609 nm can help in characterizing vinyl and understanding its absorption behaviorin the near infrared range."1580-1609 Broad PeakThis is saying that we have a protein/polyamide vinyl, alkyl alcohol dominanting the composition. This would allbe consistent with solubility and physical properties. We seem to be on track quite well here.Now for the polymer synth layer in STRONG H2SO4 (Red-purple color) non volatile1440 Methylene (1440) aliphatic hydrocarbons, OH Aromatic (1420), OH Phenolic (1420)CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.25 Human Synth Blood Compare47 Work on this with physical propeties, UVHuman blood-synth layer comparison to continue.Mid IR exploration of vinyl group with historical IR data (cerca 2015) looks to be most important. Initial peakmatch search brings up following candidates as:1. Butyl vinyl ether C6H12O2. Cyclohexyl vinyl ether C6H14OThese candidates have essentially exact matches with IR collected Sep 2015 of "CDB Protein Complex".Out of about 10 candidates from entire SDBS database. These are lowest molecular weight. I am guessing wehave something here.Look at physical properties, synthetic rubber, etc.Combination of human-synth blood comparison with polymerization probable is a worth topic.Therefore we have quite a bit of information to work with. Our main topics covered are:1. UV Spectrums compared2. NIR spectrums compared3. Expected impact upon blood4. Nature of vinyl5. Red color of polymer in H2SO4, Photos6. Photographs under scope - synth cells7. Polymer macro photographs8. We have reference MID IRs for both human blood and the "protein complex" from cerca 2015Major removal information related to this reaction (sulfuric acid + blood) has been scrubbed from AI Chat andYouTube after original inquiries 24 hours earlier. Any relationships to use of sulfuric acid in the reaction havebeen removed.So the conclusion is because this reaction is occurring with the synth polymer layer it is another level ofconfirmation of the existence of hemoglobin within the synth polymer layer. Scrubbing of information looks tobe pretty clear on this research.Now, is there anything more that we can learn with the equipment and means that we have?1. What are the vinyl groups in MID IR and is ther any sign of it within our historical MID IR blood spectra?2. Can you find any signs of the problem in historical MID IR?What should be the title of this paper?Human Blood vs. Synthetic Blood : A Comparison & a VERY Strong LeadIn addition, the more detailed vinyl identification candidate is an important topic.Look into the type of polymerization with the specific vinyl under investigation."Cyclohexyl vinyl ether is used in the production vinyl chloride polymers. In fluoropolymer resins, it contributessolubility, adhesion and crosslinking making a highly durable coating.""Cyclohexyl vinyl ether is an organic compound with the chemical formula C8H14O. It is a colorless liquid thatis used as a monomer in the production of various polymers and copolymers. It can also be used as a solventor as a reagent in organic synthesis. Cyclohexyl vinyl ether is known for its ability to undergo polymerizationreactions via radical or cationic mechanisms."CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.25 Human Synth Blood Compare48 "The vinyl group, which consists of a double bond between two carbon atoms, typically absorbs infrared (IR)radiation in the range of 1640-1680 cm^-1. This absorption is due to the stretching vibrations of thecarbon-carbon double bond."The bacteriophage is an important topic.Electrochemical measurements of organic redox reactions? That would open up a whole new ballgame.An important question arises. We now have a significant MID IR match level occurring between the "CDBProtein Complex" MID IR spectrum collected on Sep 05 2015 and the compound Cyclohexyl VInyl Etherinvolving 8 peaks. Quite strong.Now we also have a strong match between CEC collected blood spectrum of Nov 07 2015 and the spectrum ofalbumin, the primary protein in blood plasma, recorded on the CDBS.Now the question is, are there any peaks that show up on the CVE spectrum that also show up on the CECblood spectrum that do not show up on the reference MID IR listing for human blood?So the peaks on the Cyclohexyl vinyl ether spectrum that match the CDB Protein Complex of 09.05.15 are:2927, 2858, 1739, 1641, 1456, 1377, 1177, 1040.Now which of these peaks match CEC Blood Spectrum of 11.07.15?2919, 2857, 1646, 1452 and 1389.Now which of these peaks do not match my reference MID IR blood data of the notes of Nov 03 2023 (handnotes, Vol 33)? Also human blood reference work done on Oct 31 2023 (hand notes).2919 cm-1 does not show a match yet.2858 does show a match (lipids claimed).1646 does show a match (amide I C=O claimed).1452 does not show a match.1389 does show a match (lipids and proteins claimed).1042 does not show a match.Now in human blood we are apparently seeing a NIR entry of 1628 nm. This corresponds to vinyl andvinylidene (1630, 1631) and this is a huge red flag that should now be there.In addition, I am now to trace MID IR peaks of:2919 cm-1, 1452 cm-1 and 1042 cm-1. and look for any pattern of ill fit shared between MID IR and NIR.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.25 Human Synth Blood Compare49 2023.11.26 Blood Synth Vinyl MID IRI have an interesting observation regarding the purple color on the synth-polymer layer, and I need to recallwhat I did. I believe it was the following:1.A small piece of the dried synth polymer layer (a tough vinyl sure fits the bill) in about a ml of H2O. Then Ithink a drop of conc. H2SO4, maybe two or three. No purple color change takes place under these conditions ofa weaker H2SO4 solution. Put it in the frig because of something I remember reading regarding chillinginducing a purple reaction with blood and sulfur.Remembered two days later to look at it. It is indeed purple, just as I have been able to achieve with very conc.H2SO4 at rooom temperature with the synth layer.So I will be:1. Looking at this more and for replication.2. Try to find what I read regarding H2SO4, blood and chilling purple reaction.l3. Ask the question, this means that the vinyl compound/product would have a reaction similar to with bloodand therefore share another characteristic of blood and hemoglobin interaction....It also means that sulfurinteracts with the synth polymer without doubt, in a way that is similar to how sulfur can react with blood."Sulfhemoglobinemia is a rare condition characterized by the presence of sulfhemoglobin in the blood.Sulfhemoglobin is a form of hemoglobin that contains sulfur instead of iron in its heme group. This alteration inthe structure of hemoglobin affects its ability to bind and transport oxygen, leading to a decrease inoxygen-carrying capacity in the blood.The exact cause of sulfhemoglobinemia is not fully understood, but it is thought to be a result of exposure tocertain chemicals or medications. Some known triggers include certain drugs like phenacetin, sulfonamides,and aniline dyes, as well as exposure to industrial chemicals and toxins.Symptoms of sulfhemoglobinemia can vary but may include cyanosis (bluish discoloration of the skin),shortness of breath, fatigue, dizziness, and confusion. In severe cases, it can lead to tissue damage and organdysfunction.Diagnosis of sulfhemoglobinemia is typically made through a blood test that measures the levels ofsulfhemoglobin in the blood. Treatment options are limited, and there is no specific cure for the condition. Inmost cases, the condition resolves on its own once the exposure to the triggering agent is removed.Supportive care, such as oxygen therapy, may be provided to manage symptoms and improve oxygenation.It is important to consult with a healthcare professional for a proper diagnosis and management ofsulfhemoglobinemia. They can provide personalized advice and guidance based on individual circumstances.""Sulfhemoglobinemia is a rare condition in which there is excess sulfhemoglobin (SulfHb) in the blood. Thepigment is a greenish derivative of hemoglobin which cannot be converted back to normal, functionalhemoglobin. It causes cyanosis even at low blood levels.It is a rare blood condition in which the ß-pyrrole ring of the hemoglobin molecule has the ability to bindirreversibly to any substance containing a sulfur atom.[1][2] When hydrogen sulfide (H2S) (or sulfide ions) andferrous ions combine in the heme of hemoglobin, the blood is thus incapable of transporting oxygen to thetissues.PresentationSymptoms include a blueish or greenish coloration of the blood (cyanosis), skin, and mucous membranes, eventhough a blood count test may not show any abnormalities in the blood. This discoloration is caused by greaterthan 5 grams per cent of deoxyhemoglobin, or 1.5 grams per cent of methemoglobin, or 0.5 grams per cent ofsulfhemoglobin, all serious medical abnormalities.[citation needed.""Occurs when a sulfer atom binds to porphyrin ring of hemoglobin, resulting in permanent oxidation of iron tothe ferric state, incapable of oxygen transportMost often associated with phenazopyridine, dapsone, metoclopramide, sumatriptanAlso associated with industrial chemicals, including trinitrotoluene, hydroxyl amine sulfate, dimethyl sulfoxide,Hydrogen Sulfide"2023.11.26 Blood Synth Vinyl MID IR50 "Sulfhemoglobinemia is a rare condition where the blood takes on a purple color due to the presence ofsulfhemoglobin. Sulfhemoglobin is a compound formed when sulfur combines with hemoglobin, the proteinresponsible for carrying oxygen in red blood cells. This condition can occur as a result of certain medications,chemicals, or toxins, and it can lead to symptoms such as shortness of breath, fatigue, and cyanosis(bluish-purple discoloration of the skin). Treatment for sulfhemoglobinemia typically involves identifying anddiscontinuing the causative agent, and in severe cases, blood transfusion may be necessary. It is important toconsult a healthcare professional for proper diagnosis and management of this condition."It is a morbid condition.Major removal information related to this reaction has been scrubbed from AI Chat and YouTube after originalinquiries 24 hours earlier. Any relationships to use of sulfuric acid in the reaction have been removed.So the conclusion is because this reaction is occurring with the synth polymer layer it is another level ofconfirmation of the existence of hemoglobin within the synth polymer layer. Scrubbing of information looks tobe pretty clear on this research.OK, there is some very clear evidence available of vinyl in the blood. RIght now it is from NIR, we will see if itexists in MID IR afterwards. If we look at our notes of Oct 31 2023 (handwritten) we have a clear entry ofabsorbance at 1627 nm within a human blood analysis. This is clearly indicated within the CRC reference asbeing of vinyl nature from 1621 - 1637. Five consecutive entries with no real competition except for acrylate at1621. This is a sufficient entry to show crossover between synth polymer analysis and human blood.OK, we are at the point now where we can provide direct evidence of vinyl influence on human blood via NIR.The method is to focus on human blood alone, not comparing it to anything, and look at with NIR. We alsofocus in on the vinyl region of 1627 nm and remove extraneous examinations. The evidence is quite strongnow of the existence of vinyl groups, polymerization, synthetic blood, within human blood. This is from ahuman blood sample 2023, unvaccinated individual.It now becomes less important, but the question arises, can you show evidence of vinyl presence in a humanblood MID IR spectrum available in 2015? We know we can show evidence from the MID IR of a "CDB ProteinComplex" MID IR spectrum of 2015 in combination with use of SDBS database."The typical wavelength range for infrared absorption of vinyl groups is around 1600-1650 cm^-1."Caution: Amides also absorb in ths region!"The amide absorption in the infrared region of human blood typically occurs at wavelengths between 1600and 1700 cm-1. This absorption is mainly attributed to the stretching vibrations of the amide I and amide IIbonds in proteins present in the blood." TheNow we definitely have absorption of the protein complex MID IR of Sep 05 2015 at 1638 cm-1. This confirmsthe likely existence of the vinly group detected within the cultures of 2015. Now what about in blood? We haveabsorption at 1646 but so does amide."The simplest vinyl compound is vinyl chloride (C2H3Cl).""An ether functional group is a functional group in organic chemistry that consists of an oxygen atom bondedto two alkyl or aryl groups. It is represented by the formula R-O-R', where R and R' are alkyl or aryl groups.Ethers are characterized by the presence of the oxygen atom in the molecule, which gives them uniquechemical and physical properties""Ether absorption in the infrared region typically occurs in the range of 1000-1300 cm-1. This absorption is dueto the C-O stretching vibration of the ether functional group. The exact position of the absorption peak mayvary depending on the specific structure of the ether molecule."CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.26 Blood Synth Vinyl MID IR51 Now, here is what we are seeing. The 2015 CDB MID IR shows strong absorption at 1038 cm-1. THis looks likeit MAY correspond to ether.Molecules that contain a C-O bond typically absorb infrared radiation at around 1038 cm-1. Ether, which hasthe chemical formula R-O-R', where R and R' can be any alkyl or aryl group, contains a C-O bond and canabsorb infrared radiation at this frequency.I have found the MID IR correlation that I am seeking. My spectrum with albumin matches EXTREMELY WELLexcept for 2 peaks that exist in the range of ethers and vinyls. 1298 and 1160 cm-1. SDBS will find that theclosest match to these two peaks is ethyl vinyl ether, exactly the top candiate from the CDB Protein complexMID IR of 2015.Poly(vinyl cyclohexyl ether) (PVChE) is a type of polymer that is derived from vinyl cyclohexyl ether monomers.It is often used in the field of biological materials due to its unique properties and potential applications.PVChE is biocompatible, meaning it is non-toxic and does not cause harm to living organisms. This makes itsuitable for use in medical and biological applications, such as drug delivery systems and tissue engineering.PVChE can be easily synthesized and modified to achieve desired properties, such as controlled release ofdrugs or enhanced mechanical strength. It can also be combined with other polymers or materials to createcomposite materials with specific functionalities.In addition, PVChE has good solubility in various solvents, allowing for easy processing and fabrication intodifferent forms, such as films, fibers, and hydrogels. This versatility makes it a valuable material for a widerange of biological applications.Overall, PVChE is a promising material in the field of biological materials due to its biocompatibility, tunableproperties, and processability. Ongoing research and development in this area aim to further explore itspotential applications and optimize its performance in various biological systems.Health observation: Besides severe skin damage (best analogy is a year + long internal burn and bee sting(eg, formic acid)) that covers the entire lower leg, angle and foot) significant edema is in place. Swelling isaggravated further with any increase in salts such as sodium (sodium bicarbonate used in pH control iscounterproductive it seems). The observation is that ultrasound applied diligently and with requisite pain atpoint of core pain reduces the swelling temporarily in that area so that leg is normally sized. US drives the fluidaway, albeit only locally. It is already known from under the scope that US separates the blood from thepolymer.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.26 Blood Synth Vinyl MID IR52 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.26 Blood Synth Vinyl MID IR53 2023.11.27 Research Paper:Blood CompareKeep refining the flow of the paper in progress:Human Blood vs. Synthetic Blood : The Path to the Blood Clot (revised title)After intro, we are on the opening subject of microscopic comparison first.This is the general flow outline of the research paper in progress:"I also have a good macro picture of the polymer synth layer on a glass slide. So good, we have1. UV of human blood, very distinctive.2. Microphotos of synth blood. Easy to compare to human blood photos. 8000x.3. Macro photo of synth polymer layer.4. We have a NIR comparison that demonstrates the polyamide/vinyl nature of the synth blood. Likelymethylation disturbance at play.5. Mid IR of vinyl groups is coming into play6. Macro photo or culture dish surface layer.7. UV comparison spectra between human blood and synthetic blood, high correspondence.8. Photo of "blood red" color change upon mixing polymer surface layer and strong H2SO4also recall kastly meyer, hemoglobin, and pI determination.9. MID IR correlation with non fitting albumin peaks.A couple more phenomenall things have happened.1. When you take the dried synth material and put in in strong H2S04, it turns immediately deep red. Slightpurple tint. Hard to avoid the situation with blood. I have photo."In terms of lab work,running questions are:1. How does the blood clot NIR compare to the poly-synth layer NIR?THis would give important information about changes that could explain the solublility behavior (i.e., or thelack of it) This would lead to an increased understanding of the polymerization process.2. I wonder what produced the purple color in the synth layer + H2SO4 in the refrigerator? I can't duplicatethat yet.3. The bacteriophage is a huge issue.4. MIght also want to revisit the nucleic acid prospect.5. Additional enzyme and polymerization study. Methylation also continues. Mitigation pursuits.Observation: The TSP 100.7D series in petri dishes does not seem to produce anything particularly unuusual. Ithink the motive for this series was an attempt to produce the polymer-synth layer assuming that increasedsurface area of petri dishes might be more benefiicial. This did not show to be the case. Thus far, largeVOLUME of culture COMBINED with large surface area seems to be the most favorable condition for productionof the polymer synth layer. I now have two very productive cultures. Both were created with 6500 ml culturesthat were concentrated down to about 1500 ml. The containers have approx a 7-8 inch diameter. There are theseries TSP 070 and TSP 100 which are very productive with strong separation into 3 distinct layers. I willdiscard the TSP 100.7D petri dish series. This series came from Nov 12. I did decide to look under the scope.Synth cell production is most certainly there; just not anything of the magnitude of the larger culture volumes.--This is our most current status with scope work at 8000x:The microscope at 8000x is working best at:1. Height of stage q tip height with one end cut off. This step is also critical. 1/8 inch change seems to affectgreatly.2. Brightness of lower dial lamp will have moderate impact, usually between 5-10.3. The diaphragm is set at 0.3 (1/4 of what the objective says)2023.11.27 Research Paper:Blood Compare54 4. Exposure Timed 40%5. Gain 20%6. Color temp 20%7. Tint 45%8. Hue 45%9. Saturation 30%10. Brightness 95%11. Contrast 100%12. Gamma 100%Today, at 3200x, it seems as though these settings can hold. It does seem that exposure control is mostprecise howeveObservation: US is positively decreasing the level of edema when it is applied at the core afflicted region of thelower leg. It drops the skin level down in swelling approx 1/4 or so, very noticeable. All consistent with knownproperties and behavior of the polymer - blood interaction under US energy. A level of pain accompanies theprocedure, approx 15 min duration.I am now in a position to start comparing the NIR of a blood clot to the NIR of the synth polymer layer. Themain uncertainties of comparison are:1. There is some ambiguity whether the clot comes from the deceased individual or a live individual (externallyclotted). The bias is strongly towards the deceased clot which is favorable, however this will likely be repeated.2. The trend is removed from the blood clot but not from the synth-polymer layer so no magnitudes of any kindcan be considered.With that being said, here is what we start to see. The main issue here involves similarity or difference in basicstructure and solubility. One thing we do know is that the clot dissolves in NaOH and the synth polymer layerdissolves in strong H2SO4. So we know already they are very different from one another and yet thepresumption is that they originate generally from the same source material. This means, since we knowe theyhave important differences, that comparison of the clot with other forms observed will be made.NIR Comparison:Start with the blood clot:Clot: 1130 (1130) 1374 (1370) 1443 (1440) `1575 (1570, 1580)and then the synth-polymer layer:Synth-Polymer: 1195 (1190, 1200) 1520 (1500) 1594 (1590) Broad 1582-1609 (1580-1610) 1700 (1700)CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.27 Research Paper:Blood Compare55 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.27 Research Paper:Blood Compare56 2023.11.28 Research Paper Video AudioCarry forward:Keep refining the flow of the paper in progress:Human Blood vs. Synthetic Blood : The Path to the Blood Clot (revised title)After intro, we are on the opening subject of microscopic comparison first.This is the general flow outline of the research paper in progress:"I also have a good macro picture of the polymer synth layer on a glass slide. So good, we have1. UV of human blood, very distinctive.2. Microphotos of synth blood. Easy to compare to human blood photos. 8000x.3. Macro photo of synth polymer layer.4. We have a NIR comparison that demonstrates the polyamide/vinyl nature of the synth blood. Likelymethylation disturbance at play.5. Mid IR of vinyl groups is coming into play6. Macro photo or culture dish surface layer.7. UV comparison spectra between human blood and synthetic blood, high correspondence.8. Photo of "blood red" color change upon mixing polymer surface layer and strong H2SO4also recall kastly meyer, hemoglobin, and pI determination.9. MID IR correlation with non fitting albumin peaks.A couple more phenomenall things have happened.1. When you take the dried synth material and put in in strong H2S04, it turns immediately deep red. Slightpurple tint. Hard to avoid the situation with blood. I have photo."3. The bacteriophage is a huge issue.4. MIght also want to revisit the nucleic acid prospect.5. Additional enzyme and polymerization study. Methylation also continues. Mitigation pursuits.I want to start revisiting the bacteriophage prospect. This event was recorded entriely in handwritten notes ofNov 07 2023. It involved the titration of TSP 70 (original large volume culture initiated on Oct 19) with a 20 dayold culture. It was apparently titrated from a level of 3.75 to 6.2. Along the way, it appears a pI of 4.31 wasdetermined. The analyte was apparently the mid level (main volume) solution of the TSP culture,understanding that it separates over time into 3 separate layers:1. Surface synth-polymer layer (white)2. Main volume of culture solution (amber)3. Bottom precipitate layer (not examined in depth to my recollection thus far)Let's try to find this titration record to determine what the equilibrium point was. The equilibrium titration pointwas recorded at a pH of 5.26. I used 50:1 NaOH-KOH titrant and was recorded in the notes of Nov 06 2023. Icarried the pH up to 6.2 therefore.It is important to realize that this was a 20 day old culture. Notice the difference in pI from the determinationof pI of a fresh culture. Fresh culture was 5.31 as I recall (human blood proteome match) whereas the 20 dayculture main solution (i.e., some separation has taken place by then) had the pI of 4.31. Variation of pI w.r.t.time was discussed within the notes - this remains uncertain at this time.Now the culture is approx. 39 days old so conditions are somewhat different. Interestingly enough, TSP 100series is about 20 days old (started on Nov 08 2023) so we actually do have a secondary reference in the2023.11.28 Research Paper Video Audio57 matter.What happens next is that you take a UV spectrum on the 6.2 remainder. At thiws point you see a strong peakshowing up at 269 nm, apparently quite definite. It is research of this peak that leads to the bacteriophageissue. I think the issue is that proteins are undersood to to center on 280 nm. DNA understood to center on260. Now tyrosine is know to be at 274 nm and this seems to be the lowest I can find for protein issues. It justseems too far out of range at 269 to place this in the protein category. I believe the peak was also pronouncedand has a different profile than protein.This also causes the nucleic acid issue to reemerge as that UV spectrum had an apparent peak at 259 nm.Problem there was that the peak was so broad that it was more difficult to isolate with complete confidencethat it had no protein nature. But this too will eventually need to be revisited. But for now, we will continue toaddress the bacteriophage issue.Do I still have the 269 nm sample available to study? Let's see. I have:TSP 70.1D YES I HAVE IT HERE. see below.TSP 70.2 see belowTSP 70.3 see belowTSP 70.4 see below.TSP 70.5 see below.and a H2SO4 control.and this is the information that I have recorded in the spreadsheet log. Good work here, I need this.Nov 07/ 2023 TSP 70.5C (B) X.XX-1107 TSP 70.5 Centrifuged TSP 70.0 Bottom Layer TSP 70.5 Centrifuged TSP70.0 Bottom Layer Have an interest in comparing this to TSP 70.2C (B) ?-1107. NIR comparison has beenmade, in notes for Nov 07.Nov 07/ 2023 TSP 70.2C (B) X.XX-1107 TSP 70.2 Centrifuged Titrate Precipitate TSP 70 was titrated. 70.1 isdecant, 70.2 is precipitate, then centrifuged This shows itself to actually be two layers. Presumed acidic asdecant has ph delta from 3.75 to 6.20.Nov 06/ 2023 TSP 70.4S (L) X.XX-1106 TSP 70.4 Solubilized Polymer-Synth Layer in Acetone Solubility tests onPolymer-Synth Layer of TSP 70.0 Acetone Success. No testing done whatsoever. Acetone control required.Presumption is that UV should also show peak at 260. Unproven - needs to be done.Nov 06/ 2023 TSP 70.3A (S)Nov 07/ 2023 TSP 70.1D (M) 6.20-1107 TSP 70.1 Titrate Decant Titrated 20 day TSP 70 Culture Decanted Thisis what shows to apparently contain bacteriophages. Big issue here. Absorbance is at 269nm. Matchesbacteriophage UV profile extremely well. 700-1100 NIR of same sample shows the alkyl alchol w/no hydrogenbonding at 962nm.I DO HAVE IT then. 70.3A is of interest for later as it is where the 259-260 peak shows up. But the 269 nmquestion looks like it requires replication.Now this culture is dated Nov 07 2023. So this is when titration was done. THe culture was approx 20 days old.The culture is now approx 40 days old but we have it which is great. Now the difference in appearance of thisculture from the original clear decant is that it has a synth polymer layer on top now and the solution is cloudy.So it has changed as we have seen before in the decant versions. It serve as a case of interest to see how theUV spectrum may have changed.This sample size is the largest of my set and is approx 40 ml. The other residual samples are on the order of afew ml. I will extract from the mid level solution area. I may also be able to get pH information at some point,however, I do not wish to disturb or mix the synth-polymer layer.Good records were helpful here. Impossible to retrace or recollect from memory.We have another important advantage now in UV. We can record with confidence from 190 nm to 400 insteadof 235 nm to 400 using the quartz cuvettes I now have in place. A much broader and better UV picture.I have replicated the spectrum. As the plot is difficult to interpret because of the broad flat peak, I have used adifferential approach. I have two peaks that at at the zero point (first once crosses, 2nd one asymptotic:CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.28 Research Paper Video Audio58 210.3 nm257 nmThe center of the broad flat peak is approx 252 nm so thus far there is no way that it approaches the proteinsection. If anything we have to wonder again if we are in the 260 region for nucleic acids again.Our notes of Nov 21 2023 are in some detail on this issue. It is somewhat problematic no matter how you goabout it. You are considering:1. protein2. nucleic acids or DNA3. bacteriophage4. or none of these and something new.Remember you did just LOOK at it under the scope and saw small crystals and apparently what seems to beprotein.molecule wavelength of maximum absorption (nm)some more information to consider. Notice all alkenes, 1, 2, 3 bonds causess displacement like we are seeing.ethene 171buta-1,3-diene 217 (UV NIST Spectrum does not seem to match this at all, they show a peak at ~291 nm.hexa-1,3,5-triene 258 (No UV spectrum available on NIST)We have some problems here. Best we may have is NIR.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.28 Research Paper Video Audio59 2023.11.30 Another Mitigation ProspectI am recognizing the similarity between nylon and solubility of the synth-polyamide layer. They are bothsoluble in strong sulfuric acid and maybe not much else. Acetone maybe some? That suggests that they sharethe same characteristic of both being polyamides.Strong research topic here....Note this comment:(Note: "Red cell membranes have a negative charge (zeta potential) that causes red cells to repel each other. In the presence of increased positively charged plasma proteins such as fibrinogen or immunoglobulins, thenegative charge on the red cell surface is diminished, allowing red cells to stick together." Ref.hematology.org)It would not be surprising if this ends up being quite important. Now start thinking about this. You have anisoelectric point of 5.33. This is when the charges are zero, i.e., electrically neutral. What charge would theprotein have if it were at pH 7.4?"The isoelectric point (pI) of a protein is defined as the pH at which the net charge of a protein molecule iszero. Accordingly, proteins are positively charged at a pH below their pI and negatively charged at a pH abovetheir pI. The protein pI varies greatly from extremely acidic to highly alkaline values ranging from about 4.0 to12.0. Hence, pI values have long been used to distinguish between proteins in methods for protein isolation,separation, purification, crystallization, etc. Amino acid composition of a protein sequence primarily defines itspI, based on the combination of dissociation constant (pKa) values of the constituent amino acids. Out oftwenty common amino acids, two amino acids, aspartic acid, and glutamic acid, are negatively charged andthree amino acids, lysine, arginine, and histidine, are positively charged at the neutral pH, as defined by theirpKa values."https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8667598/This looks like it has our answer. The pH of the culture is on the order of 3.8. The dominant pH of the humanblood proteome is 5.3. The pH of the culture is lower than the dominant pI of the blood proteome. This meansthat the blood proteins are becoming more positively charged with the introduction of the CDB.OK I have said it in the paper in progress:"((Note: !Red cell membranes have a negative charge (zeta potential) that causes red cells to repel eachother. In the presence of increased positively charged plasma proteins such as fibrinogen or immunoglobulins,the negative charge on the red cell surface is diminished, allowing red cells to stick together.# Ref.hematology.org)!The isoelectric point (pI) of a protein is defined as the pH at which the net charge of a protein molecule iszero. Accordingly, proteins are positively charged at a pH below their pI and negatively charged at a pH abovetheir pI. The protein pI varies greatly from extremely acidic to highly alkaline values ranging from about 4.0 to12.0. Hence, pI values have long been used to distinguish between proteins in methods for protein isolation,separation, purification, crystallization, etc. Amino acid composition of a protein sequence primarily defines itspI, based on the combination of dissociation constant (pKa) values of the constituent amino acids. Out oftwenty common amino acids, two amino acids, aspartic acid, and glutamic acid, are negatively charged andthree amino acids, lysine, arginine, and histidine, are positively charged at the neutral pH, as defined by theirpKa values.#Ref. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8667598/This looks like it has our answer. The pH of the culture is on the order of 3.8 (strongly acidic). The dominant pH2023.11.30 Another Mitigation Prospect60 of the human blood proteome is 5.3. The pI of the CDB culture is determined at 5.33. The pH of the CDBculture is lower than the dominant pI of the blood proteome as well as lower than the CDB culture pI. Thismeans that the blood proteins are being exposed to positively charged proteins with the introduction of theCDB.0. This would explain increased coagulation. # CEC)"This therfore provides an additional potential mitigation strategy. Increase intake of negatively chargedproteins.Mitigation prospects:1. Protein disruption : enzymes2. Methyl Disruption : Vit B, Fatty acids?3. pH regulation (risks as well, e.g, sodium increase, polymerization, misinformation, blood pH vs urine ph)4. Ultrasound5. Negatively charge protein intake?6. Antoxidants and the usual two dozen suspects7. Balm, e.g., methyl salicylateThe blood proteome distribution (dominant pI of 5.3) is at:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7159840/"Blood plasma is a major fluid compartment in human body, with a narrow normal range of pH (7.40±0.05)12.Thus the average pH of human blood is in the middle of the trough observed for the theoretical pI distributionof blood proteome (Figure 1[A]). Whether such a coincidence is by chance or by design, the density of normalproteoforms in the pI region 7.4±0.1 is very low (ca. $20 times lower than in the region around pI 5.3±0.1)."The mitigation idea would be to increase intake of proteins with a pH greater than the dominant pI of blood,which is 5.3."Accordingly, proteins are positively charged at a pH below their pI and negatively charged at a pH above theirpI."We need the latter part of this sentence. Negatively chared amino acids are:Aspartic acid pKa = 3.7 pI = 2.87Glutamic Acid pKa = 4.1 pi = 3.08This means that these proteins are negatively charged anytime that they are in a pH environment that isabove approximately 3.0. Blood is 7.0 so it is significantly above these pI values. So we ask what arenutritional sources of aspartic and glutamic acid?"What is Aspartic Acid (Aspartate)?Definition- Aspartic acid or aspartate, also known as amino succinic acid is a non-essential amino acid that issynthesized itself in the human body through different sources of foods. It is mainly responsible forsynthesizing proteins and regulating hormones so also known as building blocks.Aspartic acid is involved in synthesizing four different amino acids as it plays a vital role in Kreb!s cycle;methionine, isoleucine, lysine, and threonine. It is an aspartate family and a proteinogenic amino acid. It is alsoa neurotransmitter.Aspartic acid is the product formed by the hydrolysis of proteins. Aspartic acid was first identified in 1868 fromlegumin in plant seeds. As aspartic acid are non-essential amino they are synthesized in the body from oxaloacetic acid that is produced during the metabolism of carbohydrates.Structure of Aspartic Acid (Aspartate)Structure of Aspartic Acid (Aspartate)Aspartic acid consists of two functional groups, one amino group is basic in nature and the other is the acidiccarboxyl group. Therefore, amino acids molecule exists as a zwitterion. Aspartic acid is alanine with one of theß hydrogens replaced by a carboxylic acid group. The pKa of the ß carboxyl group of aspartic acid in apolypeptide is about 4.0. It is a dibasic amino acid having two carboxyl groups; one on alpha carbon atom andanother on the side chain. Aspartic acid has an alpha-keto homolog. Aspartic acid is divided into two forms;L-aspartic acid and D-aspartic acid. L configuration is a more common and dominant form. L-aspartic acid isCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.30 Another Mitigation Prospect61 typically involved in the production of antibodies and is part of protein synthesis in the body which isresponsible for increasing the immune system. D-aspartic acid is not involved in protein synthesis and ismainly found in the pituitary gland and testes which is used in the regulation, synthesis, and release oftestosterone and luteinizing hormone.Sources of Aspartic Acid (Aspartate)It is found in sugar cane and sugar beets molasses, asparagus, avocado, sprout seeds, and oat flakes.Animal Sources includes: Sausage meat, Luncheon meat and wild game.Other sources includes magnesium aspartate that is salt of aspartic acid and sweeter aspartame.Physical Properties of Aspartic Acid (Aspartate)Molecular weight: 133.10White, crystalline solidsPolarAcidicHydrophilicOrthorhombic, bisphenoidal leaflets or rodsSour in tasteChemical Properties of Aspartic Acid (Aspartate)Melting point: 270ºCSolubility: 5390 mg/L at 25 ºCDensity: 1.6603 at 13 ºCLogP: -3.89pKa: 2.77 because of two carboxyl moleculeBiosynthesis of Aspartic Acid (Aspartate)Protein extraction, chemical synthesis, and enzymatic conversion are three main methods to produce asparticacid. A large number of amino acids are produced in the extraction method from the hydrolysis of protein. Inthis method, L-aspartic acid should be separated. Chemical synthesis requires high temperature and pressurein a racemic mixture producing both isomers of aspartic acid. So, enzymatic conversion is the best method forthe production of aspartic acid. Bacterial fermentation is the best for the highest yield of amino acids.Pseudomonas, Bacillus, and Proteus are considered as the main producers but E. coli and Corynebacteriumglutamacium are mostly preferred by industries.Biosynthesis of Aspartic Acid (Aspartate)Figure: Biosynthesis of Aspartic Acid (Aspartate) from Fumaric Acid. Image Source: Kurt Rosentrater (MDPI).In the 1960s, the fermentation process is developed and patented that utilizes sugar-free medium and usesfumaric acid as a sole source of carbon. Ammonia serves as a nitrogen source that is used in catalysis.Ammonia and fumaric acid are used in combination as 1:1 or 1:2 ratio. the pH of the broth is initialized to 7and that naturally increases from 8.4 to 9.6 in the initial stage that allows for the production of acid.Fermentation can be done with or without agitation for 2 to 10 days at 27-40 ºC. L-aspartic acid will besecreted and accumulate in the culture broth. Different downstream processes are available to separateL-aspartic acid from the culture broth. But in the case of batch fermentation, ion exchange resins can be usedto separate and purify L-aspartic acid followed by crystallization. L-aspartic acid can be separated by adjustingthe broth to 90 ?C and a pH of 2.8 with sulfuric acid in continuous fermentation. After the pH is adjusted to 2.8,the isoelectric point will cause L-aspartic acid to precipitate out of the solution. It is then subjected to atwo-hour incubation period at 15 ºC to induce protein crystallization. Under these conditions, L-aspartic acidsyield 95%.Production of Aspartic Acid (Aspartate)Figure: Production of L-Aspartic Acid. Image Source: Kurt Rosentrater (MDPI).Functions and Uses of Aspartic Acid (Aspartate)It is easily available multivitamins that are found in different forms as tablets, powders, and fluids.Because of its role in regulating testosterone levels, D-aspartic acid is used as increasing muscle mass.It also helps in keeping the concentrations of NADH (Nicotinamide adenine dinucleotide) high in brain cells andalso increase the mind sharpness leading to further production of neurotransmitters as well as chemicalsneeded for normal mental functioning.Can also be used for increasing fertility.It is also used to produce poly aspartic acid that is used as a fertilizer synergist.It also aids in energy production, RNA and DNA synthesis, and liver detoxification.It also helps in removing excessive toxins from the cells like ammonia.It is also used as building blocks molecules for active pharmaceutical agents.It is useful in making culture medias, detergents, fungisides and germicides."https://thechemistrynotes.com/aspartic-acid/CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.30 Another Mitigation Prospect62 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.30 Another Mitigation Prospect63 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.30 Another Mitigation Prospect64 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.11.30 Another Mitigation Prospect65 2023.12.02 Abstracts 2022 PreparedPrepared abstracts for 2022 on both carnicominstitute.org and substack sites.2023.12.02 Abstracts 2022 Prepared66 2023.12.04 Media Summaries - VinylI have completed several important summaries of the work that has been done in 2022-2023. I have made avideo as well as two audio summaries of 2022 and 2023 abstracts. All work has been prepared with AIassistance; tools are very helpful and allow work to be done which would never be done otherwise.I suspect work on the order of 3 weeks full time has been completed within about 3 days of work. That is aleverage of about 20:1. I easily could have hired a team of 3 people working for those same 3 weeks.Estimated cost: 150 per day x 3 folks = $450/day. 450 * 21 days = $94,500. Not exactly an exaggeration andit show the power of the tools and shows how work that never would have been done has been completed forabout $100 of tools (at most) and 3 days of my time. It will never be quite the same again for this particulartype of work that helps others to access and understand better.Let's get back to the paper:tell me about polyamides.Polyamides are a class of polymers that are known for their toughness, strength, and resistance to wear andtear. They are commonly used in various applications, including textiles, engineering plastics, and automotiveparts.One of the most well-known polyamides is nylon, which was first developed in the 1930s. Nylon is a syntheticpolyamide that is made from a combination of chemicals, typically derived from petroleum. It is known for itshigh tensile strength, good elasticity, and excellent resistance to abrasion and chemicals.Polyamides have a unique molecular structure that consists of repeating units linked by amide bonds. Thisstructure gives them their characteristic properties. The amide bonds create a strong and rigid polymer chain,which contributes to the strength and durability of polyamides.In addition to nylon, there are several other types of polyamides, each with its own set of properties andapplications. Some common examples include:1. Polyamide 6 (PA 6): This type of polyamide is derived from caprolactam and is widely used in the productionof fibers, such as nylon 6. It is known for its excellent mechanical properties, high melting point, and goodresistance to chemicals.2. Polyamide 66 (PA 66): PA 66 is made from a combination of adipic acid and hexamethylenediamine. It issimilar to nylon 6 but has a higher melting point, better resistance to heat, and improved mechanicalproperties.3. Polyamide 11 (PA 11): PA 11 is a bio-based polyamide derived from castor oil. It is known for its excellentresistance to chemicals, UV radiation, and hydrolysis. It is often used in applications where high-performanceand sustainability are required, such as automotive components and sports equipment.4. Polyamide 12 (PA 12): PA 12 is a flexible and tough polyamide that is commonly used in the production oftubing, pipes, and hoses. It has good resistance to chemicals, impact, and abrasion.Polyamides are versatile materials that can be modified to meet specific requirements. They can be reinforcedwith fillers, such as glass fibers, to enhance their strength and stiffness. They can also be blended with otherpolymers to create hybrid materials with unique properties.Overall, polyamides are widely used in various industries due to their excellent mechanical properties,chemical resistance, and durability. Their versatility and ability to be tailored to specific applications makethem an important class of polymers.what is the difference between amines and amides?Amines and amides are both organic compounds that contain nitrogen, but they differ in their functionalgroups and chemical properties.Amines are compounds that have a nitrogen atom bonded to one or more alkyl or aryl groups. They can beclassified as primary, secondary, or tertiary amines, depending on the number of alkyl or aryl groups attached2023.12.04 Media Summaries - Vinyl67 to the nitrogen atom. Amines are basic in nature and can act as nucleophiles in chemical reactions. They oftenhave a strong ammonia-like odor.Amides, on the other hand, are compounds that have a nitrogen atom bonded to a carbonyl group (C=O). Thecarbonyl group is typically attached to an alkyl or aryl group. Amides can be classified as primary, secondary,or tertiary, similar to amines. Unlike amines, amides are not basic and do not have a strong odor. They havehigher boiling points and are less volatile than amines.In terms of their chemical properties, amines can undergo reactions such as nucleophilic substitution,alkylation, and oxidation. Amides, on the other hand, are less reactive and undergo reactions such ashydrolysis, reduction, and dehydration.In summary, the main difference between amines and amides lies in their functional groups and reactivity.Amines have a nitrogen atom bonded to alkyl or aryl groups and are basic in nature, while amides have anitrogen atom bonded to a carbonyl group and are less reactive.can amines form a polymer?Yes, amines can form polymers. A polymer is a large molecule composed of repeating subunits calledmonomers. Amines, which are compounds that contain a nitrogen atom bonded to one or more carbon atoms,can undergo polymerization reactions to form polyamines.Polyamines are polymers that consist of repeating amine units. They can be formed by various polymerizationmethods, such as step-growth polymerization or chain-growth polymerization. In step-growth polymerization,two or more monomers react together to form a polymer, while in chain-growth polymerization, monomers addone by one to the growing polymer chain.Polyamines have a wide range of applications, including as adhesives, coatings, and flocculants. They can alsobe used in biomedical and pharmaceutical fields, as well as in the production of certain plastics and fibers.what are the chemical and physical properties differences between polyamines and polyamides?Polyamines and polyamides are two different types of polymers with distinct chemical and physical properties.Chemical Properties:1. Composition: Polyamines are polymers composed of repeating amine (-NH2) functional groups, whilepolyamides are polymers composed of repeating amide (-CONH-) functional groups.2. Reactivity: Polyamines are generally more reactive than polyamides due to the presence of primary andsecondary amine groups. They can undergo various chemical reactions such as condensation, cross-linking,and complex formation. Polyamides, on the other hand, have lower reactivity but can still undergo certainreactions like hydrolysis and esterification.3. Solubility: Polyamines are typically soluble in polar solvents like water and alcohols due to the presence ofamine groups that can form hydrogen bonds. Polyamides, however, are generally insoluble in water but candissolve in certain organic solvents like formic acid, sulfuric acid, and phenol.Physical Properties:1. Melting and Boiling Points: Polyamines generally have lower melting and boiling points compared topolyamides. This is because polyamines have weaker intermolecular forces (such as hydrogen bonding)between the polymer chains, leading to lower energy requirements for breaking these forces. Polyamides havestronger intermolecular forces, resulting in higher melting and boiling points.2. Strength and Rigidity: Polyamides are known for their high strength and rigidity, making them suitable forapplications requiring structural integrity. Polyamines, on the other hand, are generally more flexible and havelower strength due to the presence of amine groups that disrupt the polymer chain packing.3. Thermal Stability: Polyamides exhibit higher thermal stability compared to polyamines. Polyamides canwithstand higher temperatures without significant degradation due to the strong amide linkages, whereaspolyamines may undergo thermal decomposition at lower temperatures.4. Electrical Conductivity: Polyamides are generally insulators, while some polyamines can exhibit conductivitydue to the presence of amine groups that can act as charge carriers.Overall, the chemical and physical properties of polyamines and polyamides differ significantly due to theirCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl68 chemical compositions and structural arrangements. These differences make them suitable for variousapplications in industries such as textiles, coatings, adhesives, and biomedical materials.give me some examples of both polyamines and polyamides. give some commercial product examples also.Polyamines are organic compounds that contain multiple amino groups (-NH2). Here are some examples ofpolyamines:1. Ethylenediamine: It is a common polyamine with the chemical formula C2H8N2. Ethylenediamine is used asa building block in the production of various chemicals, such as nylon, pesticides, and pharmaceuticals.2. Putrescine: It is a diamine with the chemical formula C4H12N2. Putrescine is naturally found in livingorganisms and plays a role in biological processes. It is also used in the production of polyamides,pharmaceuticals, and cosmetics.3. Spermidine: It is a triamine with the chemical formula C7H19N3. Spermidine is found in all living cells and isinvolved in cell growth and differentiation. It is used in various research applications, such as cell culture andmolecular biology experiments.Polyamides, on the other hand, are polymers that contain amide (-CONH-) linkages in their backbone. Here aresome examples of polyamides:1. Nylon: Nylon is a synthetic polyamide commonly used in textiles, such as clothing, stockings, and carpets. Itis also used in the production of various other products, including ropes, fishing lines, and automotive parts.2. Kevlar: Kevlar is a high-strength polyamide known for its exceptional toughness and resistance to impact. Itis used in the production of bulletproof vests, helmets, and other protective gear. Kevlar is also used in theaerospace industry for its lightweight and high strength properties.3. Polyamide-imide (PAI): PAI is a high-performance polyamide used in applications that require excellentmechanical properties, high temperature resistance, and chemical resistance. It is used in industries such asautomotive, electronics, and aerospace for components like bearings, seals, and electrical connectors.These are just a few examples of polyamines and polyamides, and there are many other compounds andcommercial products associated with these polymer families.now tell me three common ways that synthetic rubber is made.Sure! Here are three common ways that synthetic rubber is made:1. Polymerization: This method involves the polymerization of monomers to form long chains of rubbermolecules. The most common monomers used in synthetic rubber production are butadiene and styrene.Polymerization can be achieved through various techniques such as emulsion polymerization, solutionpolymerization, or bulk polymerization.2. Copolymerization: In this approach, two or more different monomers are combined to create copolymers. Byblending different monomers, the resulting synthetic rubber can have enhanced properties compared torubber produced from a single monomer. For example, the copolymerization of butadiene and styreneproduces a type of synthetic rubber called Styrene-Butadiene Rubber (SBR), which is widely used in tiremanufacturing.3. Vulcanization: Vulcanization is a process used to improve the properties of synthetic rubber. It involves theaddition of various chemicals, such as sulfur, accelerators, and activators, to the rubber compound. The rubberis then heated, which causes the chemical additives to react and cross-link the rubber molecules. Thiscross-linking process enhances the rubber's strength, elasticity, and resistance to heat, aging, and abrasion.These are just a few common methods used in the production of synthetic rubber. The specific techniquechosen depends on the desired properties and applications of the final rubber product.is butadience a vinyl product?No, butadiene is not a vinyl product. Butadiene is a colorless gas that is used as a raw material in theproduction of synthetic rubbers, such as styrene-butadiene rubber (SBR) and polybutadiene rubber (PBR). Vinylproducts, on the other hand, are made from vinyl chloride monomer and include materials like polyvinylchloride (PVC).what are examples of polymer vinyls?CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl69 Some examples of polymer vinyls include:1. Polyvinyl chloride (PVC): PVC is a widely used polymer vinyl that is known for its versatility and durability. Itis commonly used in construction materials, pipes, electrical cables, and vinyl flooring.2. Polyvinyl acetate (PVAc): PVAc is a polymer vinyl often used in adhesives, paints, and coatings. It is also themain component in white glue or wood glue.3. Polyvinyl alcohol (PVA): PVA is a water-soluble polymer vinyl that is used in various applications such asadhesives, coatings, and textiles. It is also used in the production of PVA films and fibers.4. Polyvinylidene chloride (PVDC): PVDC is a polymer vinyl that is known for its excellent barrier propertiesagainst gases and moisture. It is commonly used in food packaging films, shrink wrap, and protective coatings.5. Polyvinylpyrrolidone (PVP): PVP is a polymer vinyl that is used in pharmaceuticals, cosmetics, and personalcare products. It is often used as a binder, film-former, or stabilizer in various formulations.These are just a few examples of polymer vinyls, and there are many more variations and derivatives availablein the market.what are the chemical and physical property differences between dienes and trienes?Dienes and trienes are both types of hydrocarbons that contain double bonds. However, there are somedifferences in their chemical and physical properties:1. Chemical Properties:- Dienes have two carbon-carbon double bonds, while trienes have three carbon-carbon double bonds.- Dienes can undergo addition reactions with electrophiles, such as hydrogenation or halogenation, at eachdouble bond independently. Trienes can also undergo similar addition reactions, but with the presence of threedouble bonds, the reactions can be more complex.- Dienes can undergo polymerization reactions to form polymers, such as synthetic rubbers. Trienes can alsoundergo polymerization, but the presence of more double bonds can lead to more complex polymer structures.- Dienes can undergo Diels-Alder reactions, which involve cycloaddition reactions with dienophiles. Trienes canalso participate in Diels-Alder reactions, but with the presence of more double bonds, the reactions can havemore diverse outcomes.2. Physical Properties:- Generally, as the number of double bonds increases from dienes to trienes, the boiling points and meltingpoints tend to increase. This is due to the increased intermolecular forces (Van der Waals forces) between themolecules.- The presence of double bonds in both dienes and trienes can lead to increased reactivity, making them moreprone to oxidation and other chemical reactions compared to saturated hydrocarbons.- Both dienes and trienes are typically liquids or low-melting solids at room temperature, depending on thespecific structure and molecular weight.It's important to note that these properties can vary depending on the specific diene or triene compound, asdifferent substituents and structural arrangements can influence their behavior.so we know that dienes and trienes are alkanes.what it is called when you have two vinyl functional groups combinedWhen two vinyl functional groups are combined, it is called a vinyl group dimerization or vinyl dimerization.Look at our notes of Nov 16 2023:I am looking at 100.8. This is the larger volume decant sample prepared for additional pI testing. It will nowalso be used and given equal priority to polymerized protein production (presumed identity). It is following thesame progression of 100.5 (also a titration decant) with:1. Increasing cloudinessCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl70 2. Lowering of pH3. Increased TDS4. Increased PPTas recorded on spreadsheet during vitals monitor today.We notice that the "vitals" in no way capture the whole story. They are only a helpful indicator but they canmiss very important events. An example is what is happening between 100.2, 100.3 and 100.4100.4 is TSP control100.2 is .04 gms enzyme in dilute TSP100.3 is .12 gms enzyme in conc. TSPThe vitals indicate possible significance with TSP 100.2 which is true and of interest since it is forming a synthcell-polymer surface layer even though lighter concentration of enzyme has been added.However, 100.3 by vitals shows nothing of particular interest. Visually, however, it is more than important,especially as it compares to 100.2 The only difference should be an increased concentration of enzyme to 0.12gms. The culture is TRANSPARENT with NO synth cell - polymer surface layer. This is a huge difference, but thevitals miss it and only observation detects it. So stay alert is the lesson....Now let's review the NIR spectrum of the borax-decant reaction from yesterday. It was presumed that it mighthave close ties with a polyvinyl compound. We did notice and are aware, however, that the borax reactionproduct was neither water soluble or flexible.Note: (1) Weak peak (3) Strong peakTabular info comes from CRC NIR handbookPeak info:983 (2) OH from water (maybe but maybe not....) (979)Phenolic OH (990)1170 (2) Alkene, polyenes (1170)1347 (2) Methyl (1360) larger delta than normal1436 (3) Aromatic amine (1432)Methylene (1440)1480 (3) Polyamide (1480)Secondary Amine (1481)Amide/Protein (1483)and in general, high absorbance in the range of 1436 - 1480; aromatic amine is strong here.Now my initial thought was that polyvinyl alcohol related compounds might show up in this NIR spectrum.However, the OH is not dominant here if at all, and the compound is not soluble. The water question can besettled and separated out by sustained drying of the sample. Phenols are moderately soluble in water. Now weknow that this compound is highly insoluble. This indicates that OH solubility is not a strong factor here. Thepolymer and protein signatures are what dominate here. For now, I will presume water over phenol in the 983nm group but will keep an eye on this.Most conductive polymers are polyenes. Polyenes are a main target of astrobiology. Polyenes share somechemical reaction properties with PVC vs PVA. PVC is C2H3Cl (notice no OH in any way). So if of a plasticnature would seem to be more likely of PVC family than PVA since we are insoluble and somewhat crystallinenature under scope and brittle.THE MATERIAL DOES NOT PASS A CONDUCTIVE TEST WITH THE OHMMETER AT THIS POINT.so 983 peak remains undecided between OH water and alkene, polyene.Now the aromatic amines, polyamides, secondary amine, and amide protein all speak of being a polymerizedCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl71 protein, especially in light of the borax reaction which has produced it.This exists as the most current interpretation of this material that has been created, a polymerized protein.Unclear at this point if the polymerization is likely to come from aromatic or amide origin, or both for thatmatter. We know that in the end a "rubberized" polymer is developed, but this can come from the other TSPsurface synth-cell polymer layer. But out of curiosity, is rubber aromatic or amide based?Rubber is alkene based. OK, this gets very interesting. Synthetic rubber is a copolymer between styrene andbutadiene. Styrene is a VINYL group attached to an aromatic ring. Butadiene is an alkene.Guess what, all the signs are starting to point the same direction. Aromatic, vinyl, alkenes are the componentsof synthetic rubber, which is most characteristic of the mature clot gross properties. Aromatic, vinyl, alkenesappear to be dominant structures of our NIR plots. Do not keep trying to force a single functional group. Thecombination of these three functional groups along with synthetic blood go a long way toward explaining theculture findings in general.Start looking at these three functional groups in a combined sense, understanding that separations in theculture will likely break these into parts. We will now keep an eye on the NIR plots more closely with thesevarious combinations.Let's create more of the proposed polymerized protein, now designated as TSP 100.10. Done. Drying.I am trying to acidify TSP 100.10 material w/ moderately strong H2SO4. It is difficult. It may be slightly solublein H2SO4 but certainly not easily. This means that both polymeric forms, TSP surface layer synth cell-polymerlayer and this newly developed polymer from the decant and borax are BOTH very much insoluble in water anddifficult to solubilize in reagents as well. Just like the clot was....I need to conduct solubility tests on 100.10 just like I did on the synth surface polymer on TSP 070. More studythere again as TSP 100.0 matures.My goal is to attempt to dissolve (hydrolize) TSP 100.10 for a UV run to determine if protein is detected.OK, that was straightforward. Definite absorbance at 275 nm. Protein confirmed. Polymeric protein remainsconfirmed. Now our goal is to try and determine the nature of the polymer. Our first NIR run says we have:1. alkenes, polyenes2. methyl possible3. aromatic amines4. methylene5. polyamide5. secondary amine6. amide/protein7. polyamide, amideTherefore, the reduced and presumed polymer set is:1. polyenes2. aromatics3. amines4. polyamideSeems like an ideal polymerized protein to me. Qualifies as a constituent of the final clotting process. Missingquality seems to be the rubber nature, but this seems to fit quite well for the TSP 070 surface synth layer -polymer.The essence of the clot seems to be established at this point:1. Synth erythrocytes w/hemoglobin2. Two polymeric forms, one for structual density and the other for elasticity. Both highly insoluble, even inmany strong reagents/acids/caustics.This is a major achievement. It would certainly be of benefit to understand how the two polymers may jointogether.Now the rubber property would be imparted by the vinyl group, which we now understand is a part of thecopolymeric process of synthetic rubber. We may well have, and are expected to have a copolymerarrangement here.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl72 This means that we would expect to see the vinyl component from one of the two polymers, and the strongestcandidate for that is now the TSP synth cell - polymer surface layer. Let's look at NIR plots.Guess what? (The database sure pays off). From notes of Nov 12, the vinyl component is in the precipitate!This is the other half of the TSP prior to titration!. We have all the parts we need. We have therefore separatedbetween:Three polymers, consistent of all needed parts:One separated polymer contains:1. polyenes2. aromatics3. amines4. polyamide(i.e., proteins and binding polymers)The second polymer, i.e. the precipitate of the titration:1. contains vinyl(i.e., imparts elasticity)and synthetic blood containing hemoglobin in a polymer matrix, also elastic in nature (thus protein & polymerhere also).This means you need to know if the precipitate contains protein also. I think NIR says yes. But let's work onthat one.This is all leading to the likely conclusion of three polymeric protein complexes:1. Decant2. Precipitate3. Surface synth cell layerEach one will have its own dominant chemical properties.This would therefore certainly be a copolymeric operation. Expected that decant + preciptitate = synth cellsurface layer with synthetic blood.The precipitate to the titration is 100.6. We have enough of this material to work with for now. It is importantto realize that this layer, upon centrifuge, actually contains two layers, an upper grey layer higher in volume,and a lower white layer minimal in volume. This layer now becomes important in seeking out the vinylcontribution to the overall polymer structure. For now we will focus only on the grey upper layer as itcomprises the bulk of the volume of centrifugation.We have it. NIR evaluation of the borax polymer vs the precipitated tells us the distribution and make up of thesynth surface layer.TSP 100.6 (the precipitate from titration) has high absorbance in the 1640 region. This is the vinyl dominantregion of NIR.TSP 100.10 (the borax polymer) has high absorbance in the 1440 region. This is dominated by aromatic amine.The precipitate is not high in protein content. 293 nm and 232 peaks were identified in the 100.6 acidifiedprecipitate.So what this is saying is that the middle layer of the culture (most volume) contributes the bulk of protein. Theprecipitate contributes the elasticity.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl73 So our three main components (everything is temperature and pH dependent for expression (~18 expressionsthus far) are:1. Synthetic blood (w/hemoglobin) (scores of proteins within blood) (surface layer of mature culture)2. A set of proteins that can be polymerized with borax, structurally firm but not elastic. (within decant)3. A vInyl contribution for elasticity of the culture. (within precipitate)This makes up the majority of the clot structure.1,3-Butadiene (a synthetic rubber) from NIST data base UV spectral search shows a peak at 293. This is a vinylcompound and then a match for our spectrum of TSP 100.6 at this point (not the rest of the spectrum nor our232 peak found.NIST and Chemspider both have some UV searching capability if you can get a part of the name. VERYVALUABLE!The styrene UV spectrum explains high absorbance towards the 232 peak. Synthetic rubber is a copolymerbetween styrene and butadiene.Also our notes of Nov 26 2023 look to be of interest:I have an interesting observation regarding the purple color on the synth-polymer layer, and I need to recallwhat I did. I believe it was the following:1.A small piece of the dried synth polymer layer (a tough vinyl sure fits the bill) in about a ml of H2O. Then Ithink a drop of conc. H2SO4, maybe two or three. No purple color change takes place under these conditions ofa weaker H2SO4 solution. Put it in the frig because of something I remember reading regarding chillinginducing a purple reaction with blood and sulfur.Remembered two days later to look at it. It is indeed purple, just as I have been able to achieve with very conc.H2SO4 at rooom temperature with the synth layer.So I will be:1. Looking at this more and for replication.2. Try to find what I read regarding H2SO4, blood and chilling purple reaction.l3. Ask the question, this means that the vinyl compound/product would have a reaction similar to with bloodand therefore share another characteristic of blood and hemoglobin interaction....It also means that sulfurinteracts with the synth polymer without doubt, in a way that is similar to how sulfur can react with blood."Sulfhemoglobinemia is a rare condition characterized by the presence of sulfhemoglobin in the blood.Sulfhemoglobin is a form of hemoglobin that contains sulfur instead of iron in its heme group. This alteration inthe structure of hemoglobin affects its ability to bind and transport oxygen, leading to a decrease inoxygen-carrying capacity in the blood.The exact cause of sulfhemoglobinemia is not fully understood, but it is thought to be a result of exposure tocertain chemicals or medications. Some known triggers include certain drugs like phenacetin, sulfonamides,and aniline dyes, as well as exposure to industrial chemicals and toxins.Symptoms of sulfhemoglobinemia can vary but may include cyanosis (bluish discoloration of the skin),shortness of breath, fatigue, dizziness, and confusion. In severe cases, it can lead to tissue damage and organdysfunction.Diagnosis of sulfhemoglobinemia is typically made through a blood test that measures the levels ofsulfhemoglobin in the blood. Treatment options are limited, and there is no specific cure for the condition. Inmost cases, the condition resolves on its own once the exposure to the triggering agent is removed.Supportive care, such as oxygen therapy, may be provided to manage symptoms and improve oxygenation.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl74 It is important to consult with a healthcare professional for a proper diagnosis and management ofsulfhemoglobinemia. They can provide personalized advice and guidance based on individual circumstances.""Sulfhemoglobinemia is a rare condition in which there is excess sulfhemoglobin (SulfHb) in the blood. Thepigment is a greenish derivative of hemoglobin which cannot be converted back to normal, functionalhemoglobin. It causes cyanosis even at low blood levels.It is a rare blood condition in which the ß-pyrrole ring of the hemoglobin molecule has the ability to bindirreversibly to any substance containing a sulfur atom.[1][2] When hydrogen sulfide (H2S) (or sulfide ions) andferrous ions combine in the heme of hemoglobin, the blood is thus incapable of transporting oxygen to thetissues.PresentationSymptoms include a blueish or greenish coloration of the blood (cyanosis), skin, and mucous membranes, eventhough a blood count test may not show any abnormalities in the blood. This discoloration is caused by greaterthan 5 grams per cent of deoxyhemoglobin, or 1.5 grams per cent of methemoglobin, or 0.5 grams per cent ofsulfhemoglobin, all serious medical abnormalities.[citation needed.""Occurs when a sulfer atom binds to porphyrin ring of hemoglobin, resulting in permanent oxidation of iron tothe ferric state, incapable of oxygen transportMost often associated with phenazopyridine, dapsone, metoclopramide, sumatriptanAlso associated with industrial chemicals, including trinitrotoluene, hydroxyl amine sulfate, dimethyl sulfoxide,Hydrogen Sulfide""Sulfhemoglobinemia is a rare condition where the blood takes on a purple color due to the presence ofsulfhemoglobin. Sulfhemoglobin is a compound formed when sulfur combines with hemoglobin, the proteinresponsible for carrying oxygen in red blood cells. This condition can occur as a result of certain medications,chemicals, or toxins, and it can lead to symptoms such as shortness of breath, fatigue, and cyanosis(bluish-purple discoloration of the skin). Treatment for sulfhemoglobinemia typically involves identifying anddiscontinuing the causative agent, and in severe cases, blood transfusion may be necessary. It is important toconsult a healthcare professional for proper diagnosis and management of this condition."It is a morbid condition.Major removal information related to this reaction has been scrubbed from AI Chat and YouTube after originalinquiries 24 hours earlier. Any relationships to use of sulfuric acid in the reaction have been removed.So the conclusion is because this reaction is occurring with the synth polymer layer it is another level ofconfirmation of the existence of hemoglobin within the synth polymer layer. Scrubbing of information looks tobe pretty clear on this research.OK, there is some very cliear evidence available of vinyl in the blood. RIght now it is from NIR, we will see if itexists in MID IR afterwards. If we look at our notes of Oct 31 2023 (handwritten) we have a clear entry ofaborbance at 1627 nm within a human blood analysis. This is clearly indicated within the CRC reference asbeing of vinyl nature from 1621 - 1637. Five consecutive entries with no real competition except for acrylate at1621. This is a sufficient entry to show crossover between synth polymer analysis and human blood.OK, we are at the point now where we can provide direct evidence of vinyl influence on human blood via NIR.The method is to focus on human blood alone, not comparing it to anything, and look at with NIR. We alsofocus in on the vinyl region of 1627 nm and remove extraneous examinations. The evidence is quite strongnow of the existence of vinyl groups, polymerization, synthetic blood, within human blood. This is from ahuman blood sample 2023, unvaccinated individual.It now becomes less important, but the question arises, can you show evidence of vinyl presence in a humanblood MID IR spectrum available in 2015? We know we can show evidence from the MID IR of a "CDB ProteinComplex" MID IR spectrum of 2015 in combination with use of SDBS database.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl75 "The typical wavelength range for infrared absorption of vinyl groups is around 1600-1650 cm^-1."Caution: Amides also absorb in ths region!"The amide absorption in the infrared region of human blood typically occurs at wavelengths between 1600and 1700 cm-1. This absorption is mainly attributed to the stretching vibrations of the amide I and amide IIbonds in proteins present in the blood." TheNow we definitely have absorption of the protein complex MID IR of Sep 05 2015 at 1638 cm-1. This confirmsthe likely existence of the vinly group detected within the cultures of 2015. Now what about in blood? We haveabsorption at 1646 but so does amide."The simplest vinyl compound is vinyl chloride (C2H3Cl).""An ether functional group is a functional group in organic chemistry that consists of an oxygen atom bondedto two alkyl or aryl groups. It is represented by the formula R-O-R', where R and R' are alkyl or aryl groups.Ethers are characterized by the presence of the oxygen atom in the molecule, which gives them uniquechemical and physical properties""Ether absorption in the infrared region typically occurs in the range of 1000-1300 cm-1. This absorption is dueto the C-O stretching vibration of the ether functional group. The exact position of the absorption peak mayvary depending on the specific structure of the ether molecule."Now, here is what we are seeing. The 2015 CDB MID IR shows strong absorption at 1038 cm-1. THis looks likeit MAY correspond to ether.Molecules that contain a C-O bond typically absorb infrared radiation at around 1038 cm-1. Ether, which hasthe chemical formula R-O-R', where R and R' can be any alkyl or aryl group, contains a C-O bond and canabsorb infrared radiation at this frequency.I have found the MID IR correlation that I am seeking. My spectrum with albumin matches EXTREMELY WELLexcept for 2 peaks that exist in the range of ethers and vinys. 1298 and 1160 cm-1. SDBS will find that theclosest match to these two peaks is ethyl vinyl ether, exactly the top candiate from the CDB Protein complexMID IR of 2015.Poly(vinyl cyclohexyl ether) (PVChE) is a type of polymer that is derived from vinyl cyclohexyl ether monomers.It is often used in the field of biological materials due to its unique properties and potential applications.PVChE is biocompatible, meaning it is non-toxic and does not cause harm to living organisms. This makes itsuitable for use in medical and biological applications, such as drug delivery systems and tissue engineering.PVChE can be easily synthesized and modified to achieve desired properties, such as controlled release ofdrugs or enhanced mechanical strength. It can also be combined with other polymers or materials to createcomposite materials with specific functionalities.In addition, PVChE has good solubility in various solvents, allowing for easy processing and fabrication intodifferent forms, such as films, fibers, and hydrogels. This versatility makes it a valuable material for a widerange of biological applications.Overall, PVChE is a promising material in the field of biological materials due to its biocompatibility, tunableproperties, and processability. Ongoing research and development in this area aim to further explore itspotential applications and optimize its performance in various biological systems.Health observation: Besides severe skin damage (best analogy is a year + long internal burn and bee sting(eg, formic acid)) that covers the entire lower leg, angle and foot) significant edema is in place. Swelling isaggravated further with any increase in salts such as sodium (sodium bicarbonate used in pH control iscounterproductive it seems). The observation is that ultrasound applied diligently and with requisite pain atpoint of core pain reduces the swelling temporarily in that area so that leg is normally sized. US drives the fluidaway, albeit only locally. It is already known from under the scope that US separates the blood from thepolymer.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl76 "Polystyrene with DVB: Cross-linking between polymer chains can be introduced into polystyrene bycopolymerizing with p-divinylbenzene (DVB). DVB has vinyl groups (-CH=CH2) at each end of its molecule,each of which can be polymerized into a polymer chain like any other vinyl group on a styrene monomer.""Addition polymers from conjugated dienesPolymers from conjugated dienes usually give elastomer polymers having rubber-like properties.Table 2. Addition homopolymers from conjugated dienesMonomer Polymer name Trade name UsesH2C=CH-C(CH3)=CH2 polyisoprene natural or some synthetic rubber applications similar to natural rubberH2C=CH-CH=CH2 polybutadiene polybutadiene synthetic rubber select synthetic rubber applicationsH2C=CH-CCl=CH2 polychloroprene Neoprene chemically-resistant rubberAll the monomers from which addition polymers are made are alkenes or functionally substituted alkenes."Recall the outline of our paper: (life is quite complex here):Outline of our paper:I also have a good macro picture of the polymer synth layer on a glass slide. So good, we have1. UV of human blood, very distinctive.2. Microphotos of synth blood. Easy to compare to human blood photos. 8000x.3. Macro photo of synth polymer layer.4. We have a NIR comparison that demonstrates the polyamide/vinyl nature of the synth blood. Likelymethylation disturbance at play.5. Mid IR of vinyl groups is coming into play6. Macro photo or culture dish surface layer.7. UV comparison spectra between human blood and synthetic blood, high correspondence.8. Photo of "blood red" color change upon mixing polymer surface layer and strong H2SO4also recall kastly meyer, hemoglobin, and pI determination.9. MID IR correlation with non fitting albumin peaks.A couple more phenomenall things have happened.1. When you take the dried synth material and put in in strong H2S04, it turns immediately deep red. Slightpurple tint. Hard to avoid the situation with blood. I have photo.2. In addition, there is significant overlap in the UV spectrum with human blood, especially from 190 nm to 300nm. Peaks, valleys and general profile coincides as well.Also you have a recollection of a compound which is "essentially two vinyl groups joined together"but you cannot find it.what was it?I FOUND IT! GOOD WORK.It is butadiene.Now what we need to to do is establish a pattern of likely behavior:vinly - polymer - butadienerubber componentspolyamide - sythetic bloodstyreneCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl77 combinations.polyamindes dissolving in surfuric acidspolyamides, nylon, protein differences, sulfuric acid, formic acidis butadiene a vinyl?No, butadiene is not a vinyl. Butadiene is a colorless gas that is used in the production of synthetic rubber,while vinyl refers to a type of plastic that is made from vinyl chloride monomers.So no wonder it was and is confusing. Butadiene is not technically a vinyl since "vinyl" is a particular form ofplastic. But VINYL FUNCTIONAL GROUPS make up butadiene, and even though it is a gas, it is consideredessentially as two vinyl groups joined by a single carbon bond. So from my perspective, it is definitely a vinlycompound. Semantics going on here..."1,3-Butadiene is a simple conjugated diene with the formula C4H6and can be viewed structurally as two vinyl groups ( CH2=CH2) joined together with a single bond."is butadiene a polymer?No, butadiene is not a polymer. It is a small molecule that is commonly used as a monomer to producesynthetic rubbers such as polybutadiene, which is a polymer.is triene a polymer?look into all spectroscopy matches involving butadiene, ethers vinyl...look at the mid ir matches again.separate between amines and amides, polyamides, etc.No, triene is not a polymer. Triene refers to a molecule that contains three double bonds. It is a type ofunsaturated hydrocarbon.You have a lot of data now, you need to synthesize it into a composite picture that demonstrates a polymericprogression.I have mapped out a 7 way grid to collect data from different culture states. WIll within that grid to try andsynthesize information and transfer here as it develops.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.04 Media Summaries - Vinyl78 2023.12.05 Blood Clot Progession StudyI have a ton of data here to try and assimilate and synthesize. I have to get a handle on to approach it. Wehave 4 main steps:1. The native culture characteristics, as best we can determine.2. The incubated culture where it separates into 3 layers. Top, middle, bottom.3. More detailed analysis of the three layers with an eye as to how the both separate and have combined.4. A projection into the blood clot.Let's start by trying to get a handle on the native culture. The first standard culture was TSP 070, created onOct 11 2023. It still exists and is quite mature now, almost two months old.NIR study will be a mainstay of course, maybe some UV. Some solubility. We know highly affected by pH inaddition to time separation.Let's collect what we have on relevant NIR data.Looks like our 1st NIR plot involving TSP 70 series was on Nov 07. So it was a month old by the time we startedlooking at it that way, apparently.This project of separation is looking very difficult between discerning the original native culture vs the timeseparated culture. I think we are going to have to focus on everything that is collected and look primarily forrepetion, either in TSP 070 or TSP 100.TSP 070 came into existence on Oct 08. Our studies immediately afterwards primarily involved titration and pIstudies, trying to learn about the acidic nature of the culture.But it looks like our first data came in on Oct 28 regarding functional groups. So it was about 20 days old. Nowwhat happened here is I was exploring the use of chromatography. Surprisingly it may have been moreinsightful than I knew at the time. I was working with the decant so this means the main volume AFTERprecipitation by pH.UV:274 nm Protein (best estimate is tyrosine as a dominant amino acid). HIghly soluble so the OH group makessense here.315 nm UNKNOWN - repeated later?NIR:1033 Polyfunctional alkyl alcohols (1029) Ethers and Esters also containing alcohols (we know everything isstill soluble her4e).963 Alkyl alcoholsNow what we see is that on Nov 12 BOTH TSP 100 and TSP 070, regardless of incubation time both displayedabsorbance at ~315 nm. So yes, we do have a repeat. But we do not know what 315 refers to.Ideas are:1. Synthesized metal oxide nanoparticles2. Carbohydrates in H2SO4 but we are not in H2SO4 as far as we know.Oct 28 2023So what comes out of the first functional group analysis of TSP middle layer is:1. Protein (with increased interest upon tyrosine).2. Alkyl alcohols3. Ether and Ester Alcohols4. Calcium ions (from a water test).5. Interest in metallic nanoparticles (315 nm).6. Acidic (pH is ~3.8)Two references do indicate nanoparticles at 315nm.On Oct 29 we make a NIR run with the carbonate precipitate from the decant.There certainly is no purported "AI" that can handle this problem that lies ahead. Synthesizing informationfrom widely disparate sources and methods is required. Will need to plug away one day at a time to try toextract the relevant info. Ranking is going to end up being very important so we need to identify the factorsahead of time as much as possible. Considerations are:2023.12.05 Blood Clot Progession Study79 1.age of sample2. type of sample3. solubility of sample4. pH of sample if possible, or direction towards biological pH of 7.4Let's see if I can enter the incoming information into a spreadsheet so I do not have to enter it twice.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.05 Blood Clot Progession Study80 2023.12.06 Functional Group WorksheetI am getting a spreadsheet-worksheet set up to start approaching this problem. Lots of info to synthesize. Ihave one advantage already, I can tally up in real time as I go the frequencies of functional groups appearing.Then I will work in a system for ranking including the variables of age, pH, solubility, etc. I have to get it rightto begin with so I do not have to repeat myself. I will need to go through each daily lab notes starting from Oct06 (TSP 070 creation) to the present day. This is going to take some work, but it give me the info I need.The goal is to see what functinal group and chemical progression exists from the native culture > syntheticblood > clot exists based upon the lab data.Retracing the collected information day by day has brought up an interesting observation for the notes of Nov03 2023 where human blood was compared again refernce values in the MID IR literature and synth blood andhuman blood were contemporaneously compared vi NIR.I had made a note of 1296 cm-1 being measured and not fitting anywhere in the reference values. I thendismissed that as a fluke. I see not that the measurement was not made accurately and that the actual valuefor the Aug 16 2015 human blood spectrum was at 1310 cm-1. In addition, I see another absorbance peak at1167 cm-1.I consulted 3 different reference sources. The 1310 is now actually explained by reference source no. 2 asprotein so this is now accepted. 1167 cm-1 still exists as a valid measurement and I have no curent match.It looks like 1167 does match with a potential vinyl group.My notes of Nov 26 2023 are rather of interest. This takes some sorting out. Here are the notes, which aresubsequently to be adjusted:"It now becomes less important, but the question arises, can you show evidence of vinyl presence in a humanblood MID IR spectrum available in 2015? We know we can show evidence from the MID IR of a "CDB ProteinComplex" MID IR spectrum of 2015 in combination with use of SDBS database."The typical wavelength range for infrared absorption of vinyl groups is around 1600-1650 cm^-1."Caution: Amides also absorb in ths region!"The amide absorption in the infrared region of human blood typically occurs at wavelengths between 1600and 1700 cm-1. This absorption is mainly attributed to the stretching vibrations of the amide I and amide IIbonds in proteins present in the blood." TheNow we definitely have absorption of the protein complex MID IR of Sep 05 2015 at 1638 cm-1. This confirmsthe likely existence of the vinly group detected within the cultures of 2015. Now what about in blood? We haveabsorption at 1646 but so does amide."The simplest vinyl compound is vinyl chloride (C2H3Cl).""An ether functional group is a functional group in organic chemistry that consists of an oxygen atom bondedto two alkyl or aryl groups. It is represented by the formula R-O-R', where R and R' are alkyl or aryl groups.Ethers are characterized by the presence of the oxygen atom in the molecule, which gives them uniquechemical and physical properties""Ether absorption in the infrared region typically occurs in the range of 1000-1300 cm-1. This absorption is dueto the C-O stretching vibration of the ether functional group. The exact position of the absorption peak mayvary depending on the specific structure of the ether molecule."Now, here is what we are seeing. The 2015 CDB MID IR shows strong absorption at 1038 cm-1. THis looks likeit MAY correspond to ether.Molecules that contain a C-O bond typically absorb infrared radiation at around 1038 cm-1. Ether, which hasthe chemical formula R-O-R', where R and R' can be any alkyl or aryl group, contains a C-O bond and canabsorb infrared radiation at this frequency.2023.12.06 Functional Group Worksheet81 I have found the MID IR correlation that I am seeking. My spectrum with albumin matches EXTREMELY WELLexcept for 2 peaks that exist in the range of ethers and vinys. 1298 and 1160 cm-1 [NOTE BELOW, THESE ARENOW SUPERCEDED WITH 1310 CM-1 AND 1167 CM-1]. SDBS will find that the closest match to these two peaksis ethyl vinyl ether, exactly the top candiate from the CDB Protein complex MID IR of 2015."First off, my MID IR sample here (Aug 16 2015) is actually blood serum, not whole blood. So Albumin is actuallyour best reference. Two peaks remain of interest. 1310 cm-1 and 1167 cm-1. Any measurement in notebookson Nov 03 2023 about a peak of 1296 cm-1 are now superceded with 1310 cm-1 measurement; the pick wasnot identified accurately in our spectrum. So two peaks remain in interest, 1310 cm-1 and 1167 cm-1.Next, I have MID IR reference spectrum for Albumin. These two peaks simply do not fit the spectrum. Myadditional peaks did match and they indeed are what caused albumin to show up in the list.However, a more focused search on those particular peaks led to the strongest candidate to be ethyl vinylether as mentioned above. I now remain curious about this because of the correlation to the CBB ProteinComplex MID IR Spectrum. It is therefore now time to revisit the SDBS search using these two peak values andsee if ethyl vinyl ether remains a viable candidate.Now 1 of 3 reference sources did assign a 1308 cm-1 absorption to a protein, but the other two did not. Let'slook at SDBS. Blood and serum have minor differences here. We will use blood as the question is whether avinyl group unexpectedly shows up in the blood, even years ago.OK, we have an important level of confirmation going on here on vinyl groups in the blood, back as far as 2015and 2017, two different samples. The average two peaks of interest are 1298 and 1160 cm-1. WIth searchspecs of 1 (that's one!) cm-1 and threshold of 71%, the closest match in the entire database is indeed againethyl vinyl ether. There is no requirement that vinyl be included as a condition in the search, the peaks aloneare justification for this compound. This is a very high level of confirmation of vinyl groups in blood.Strong acids dissolve nylon (polyamides).Synth-polymer of TSP dissolves in strong H2SO4.Clot dissolves in KOH-NaOH as I recall.Solubiiity tests took place on Nov 04 2023. Also alcohol detection methods took place on Nov 04 2023.Health observation. WIth sustained use of ultrasound am drawing polymer materials from my lower leg/ankle.Sustained project over a year long. Painful with much patience and perseverance required. As the ankle/lowerleg improves slowly, the material is now moving lower into the foot. Noticeable "edema" is more frequent, butthis is not just water. It will be the water soluble protein/polymer complex. Theory is that whatever can comeout of the skin represents an improvement. Be prepared for the long haul.On Nov 05 2023 the work took a strong turn toward the prospect of nucleic acids within the polmer synthmaterial. I have strong well defined absorbance at 260 nm. Will focus on the profile of the curve here also.Chilled alcohol test failed, but do nucleic acids behave the same. You also had hydrolyzed the polymer synthmaterial with strong H2SO4.Notes of Nov 12 show influence of L. enzyme upon functional groups of culture. All influences appear positive;polyamide,vinyl, aliphatic hydrocarbons, and halogenation.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.06 Functional Group Worksheet82 2023.12.07 Functional Group WorksheetContinuing to collect the data, one page, one day at a time. Now on Nov 13, 2023 of handwritten notes. Amaccumulating a detailed analysis of acquired chemical knowledge from the culturing process. The startingpoint is when the culture process stabilized from the development of TSP 070 onwards. Two large cultures arein active monitoring and use. I will attempt to provide a summary view of the worksheet results as I completethe process.Switch to the digital log is starting to take place around Nov 14 2023.Complexity of data management isacknowledged.On Nov 14 2023 the probability model developed was specified again:c = (1/range) * tan ((Problevel * pi) / 200.)Problevel = (200 / pi) * arctan(c * deltax)Very elegant to use.This was developed in radians, there is no need to convert anything. Problevel is 0-100 (i.e, percent, e.g, 70.1).Range is (e.g, RMS). Delta x is measured sample value.Have been adopting the RMS value as the reference range now at 70.% (.707). This takes care of sign andcentering issues. Very easy to implement. Peak value = 1.414 * Problevel."The form factor of an AC waveform is the ratio of its RMS value divided by its average value. Square-shapedwaveforms always have crest and form factors equal to 1, since the peak is the same as the RMS and averagevalues. Sinusoidal waveforms have an RMS value of 0.707 (the reciprocal of the square root of 2) and a formfactor of 1.11 (0.707/0.636)."Factor of 2 comes from peak to peak.In our case here, however, it does look like using the range at a 95% level of confidence is very suitable. Wecan then cut off values at >= 80% (or whatever level is desired, for example).Nov 16 2023 has a full switchover to the digital log. Doubtful that I see a return to handwritten notes exceptfor computational or diagram work, etc. required.Nov 16 Notes have the following:We have it. NIR evaluation of the borax polymer vs the precipitated tells us the distribution and make up of thesynth surface layer.TSP 100.6 (the precipitate from titration) has high absorbance in the 1640 region. This is the vinyl dominantregion of NIR.TSP 100.10 (the borax polymer) has high absorbance in the 1440 region. This is dominated by aromatic amine.The precipitate is not high in protein content. 293 nm and 232 peaks were identified in the 100.6 acidifiedprecipitate.Recall that TSP 100.6 is:TSP 100.6C (B) X.XX-1110 Centrifuged Precipitate TSP 100.0 post TitrationTwo layers grey/white centrifuge precipitate, grey dominatesWhat interests me here is the UV 293 nm and 232 peaks were not expounded upon. What might these tell us?I actually did make some comments, very interesting comments:2023.12.07 Functional Group Worksheet83 1,3-Butadiene (a synthetic rubber) from NIST data base UV spectral search shows a peak at 293. This is a vinylcompound and then a match for our spectrum of TSP 100.6 at this point (not the rest of the spectrum nor our232 peak found.NIST and Chemspider both have some UV searching capability if you can get a part of the name. VERYVALUABLE!The styrene UV spectrum explains high absorbance towards the 232 peak. Synthetic rubber is a copolymerbetween styrene and butadiene.We also know that we have this information regarding the 232 region and protein folding:"Revisiting absorbance at 230 nm as a protein unfolding probe"https://www.sciencedirect.com/science/article/pii/S0003269709001894Now this is important. poly Butadiene ALONE is a synthetic rubber, it does not need styrene."Polybutadiene - WikipediaPolybutadiene rubber is a polymer formed from the polymerization of the monomer 1,3-butadiene.Polybutadiene has a high resistance to wear and is used especially in the manufacture of tires, whichconsumes about 70% of the production."Styrene butadiene is just another format of synthetic rubber that brings in the aromatic:"Styrene-butadiene rubber (SBR) | Britannicastyrene-butadiene rubber (SBR), a general-purpose synthetic rubber, produced from a copolymer of styreneand butadiene. Exceeding all other synthetic rubbers in consumption, SBR is used in great quantities inautomobile and truck tires, generally as an abrasion-resistant replacement for natural"Yes, indeed butadiene has what is needed. It is regarded as two vinyl groups joined together. That iseverything we need to form synthetic rubber. It is all falling into place. The styrene is now just a variation to beconsidered, it makes a stronger rubber."Butadiene1,3-Butadiene is the organic compound with the formula CH2=CH-CH=CH2. It is a colorless gas that is easilycondensed to a liquid. It is important industrially as a precursor to synthetic rubber. The molecule can beviewed as the union of two vinyl groups. It is the simplest conjugated diene. Wikipedia""What have golf balls, tyres and Lego got in common?They are all made from polymers, from which the main starting material is butadiene.""1,3-Butadiene (/?bju?t?'da?i?n/)[8] is the organic compound with the formula CH2=CH-CH=CH2. It is acolorless gas that is easily condensed to a liquid. It is important industrially as a precursor to synthetic rubber.The molecule can be viewed as the union of two vinyl groups. It is the simplest conjugated diene.Although butadiene breaks down quickly in the atmosphere, it is nevertheless found in ambient air in urbanand suburban areas as a consequence of its constant emission from motor vehicles.[9]The name butadiene can also refer to the isomer, 1,2-butadiene, which is a cumulated diene with structureH2C=C=CH-CH3. This allene has no industrial significance."and the answer is yes, both sulfuric acid and HCL can dissolve tires. So solubility tests are therefore quiteimportant, including a revisit on the clot situation."What Will Dissolve Tire Rubber?There are many chemicals that can dissolve tire rubber, but the most common and effective is sulfuric acid.When mixed with water, sulfuric acid creates a highly corrosive solution that can quickly break down rubber.Other strong acids like hydrochloric acid and nitric acid can also dissolve tire rubber, but they are not ascommonly used because they are more dangerous to handle."The chain is all falling into place, isn't it?...CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet84 This was our solubiltiy information obtained on Nov 18 2023 and corresponding comments:A few solubility tests have been run on the borax polymer today. Results are:1. in strong acid (H2SO4), not soluble2. in strong alkaline (NaOH-KOH), slowly soluble3. in acetone, not soluble4. in ethanol, not solubleThese are helpful results. It has not been published yet, however, the deceased blood clot was eventuallydissolved in NaOH-KOH solution over a period of ~ 2 months. This was recorded on Aug 11 and Aug 13 2023 inVol 30 of the laboratory notebooks.This now becomes important as we have similar solubity behavior between the deceased blood clot (the mostdense and developed, rubber polymer properties) and the borax polymer that has been developed. We knowthat KOH-NaOH has generally been our only successful reagent for dissolution of any of the filaments. Weknow they help to break down the disulfide bonds, as in hair.A question is what other types of polymer bonds might they break down? In this case borate bonds areinvolved. So we have a good question coming up with the polyvinyl-borax polymer that can be made (the"slime" case). Will KOH-NOH work on that as well? If so, it shows it is effective on the polyvinyl bonds andborax, not just disulfide.Yes, lye does break down slime. YT video demo:https://www.youtube.com/watch?v=abJ1ETlhZ-wHe needed add some water instead of so much direct powder lye, but it still demonstrates the point. Waterwas used at the end and more clearly shows that breakdown has occurred.This is valuable as we are not dealing with disulfide bonds here. Vinyl bonds are involved, as they are in ourcase also.This was a valuable trial here. Another linkage in bond disruption between the clot polymer and one of theculture polymer forms.Here is some progression information that I obtained, also from Nov 18 2023I have a very significant plot now to look at. It is a NIR comparison of TSP 100.5 (Decant) over a 12 dayincubation period. This is from the time that it is decanted (amber color) until it turns white approx 10 dayslater.Main feature I am looking at is that appears there is a bulk of mass (absorbance) shift from the Nov 06 TSP100.5 (amber) at a very broad peak of ~1620 +/- 20 nm to a more focused peak at Nov 18 TSP 100.5 (White)of 1516. They do however, continue to share a great deal, and in fact the majority of the turf from 1530 - 1650nm.Interpretation of this is:1620 is alkene and vinyl1520 is protein, 1515 is polyamidethis makes a lot of sense. It is saying it starts out stronger in the vinyl section but then shifts over to a proteinnature. However, it still retains the vinyl nature. This means that a conglomerate is being formed, i.e. apolymerized protein as your surmise.This is exactly what you envision happening. This answers a very important question on culture dynamics.Let' see if this plot has been addressed yet. No, it has not, another important addition.Am looking at the reaction between baking soda and citric acidand wish to also contrast it withbaking soda + (either calcium citrate, magnesium citrate, or sodium citrate, for example)well, baking soca + citric acid creates sodium citrate + co2 gas.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet85 The reaction is: H3C6H5O7(aq) + 3 NaHCO3(s) ? 3 CO2(g) + 3 H2O(l) + Na3C6H5O7(aq)Notice that 3 moles of sodium bicarbonate are used to produce 1 mole of sodium citrate.(Think about mixing magnesium sulfate + citric acid...)"Sodium citrate is the trisodium salt of citric acid. It has a role as a flavouring agent and an anticoagulant. Itcontains a citrate(3-).ChEBISodium citrate is the sodium salt of citric acid. It is white, crystalline powder or white, granular crystals, slightlydeliquescent in moist air, freely soluble in water, practically insoluble in alcohol. Like citric acid, it has a sourtaste. From the medical point of view, it is used as alkalinizing agent. It works by neutralizing excess acid inthe blood and urine. It has been indicated for the treatment of metabolic acidosis.DrugBankSodium Citrate is the sodium salt of citrate with alkalinizing activity. Upon absorption, sodium citratedissociates into sodium cations and citrate anions; organic citrate ions are metabolized to bicarbonate ions,resulting in an increase in the plasma bicarbonate concentration, the buffering of excess hydrogen ion, theraising of blood pH, and potentially the reversal of acidosis. In addition, increases in free sodium load due tosodium citrate administration may increase intravascular blood volume, facilitating the excretion ofbicarbonate compounds and an anti-urolithic effect."This is the important aspect of what we are after:organic citrate ions are metabolized to bicarbonate ions, resulting in an increase in the plasma bicarbonateconcentration, the buffering of excess hydrogen ion, the raising of blood pH, and potentially the reversal ofacidosis.The strategy is to create citrate ions without increasing the sodium intake in a signiificant fashion.the creation of sodium citrate, somewhat acts as the precursor to the reaction:baking soda + magnesium citrate >here is a response (search was on carbonate citrate reaction, since nothing came up strong with the above, asif there is no signfiicant reaction involved. But once calcium is part of the equation, this is of interest. It says tome the process is counteproductive, and that baking soda + citric acid is where the interest lies, not bakingsoda + a citrate form:Why does Calcium (specifically Calcium Carbonate) inhibit the Citrate buffer system?Is it because citrate chelates calcium and then is unable to react with other chemicals being added? If so cansomeone explain this reaction to me, or if not provide what the reason is? If anyone has any studies they'vefound on it that would be incredibly helpful! Thank you!"Inhibited" is not probably the right term, as there is no catalyst inhibition, affecting the reaction kinetics.If we consider about neutral pHrange, the citrate buffer keeps pHby equilibrium reactionsHA2-?--?H++A3-HA2-+OH-?--?H2O+A3-with pKa3=6.4Carbonate as a base spends the buffer capacityCO32-+2HA2-?--?CO2(g)+H2O+2A3-If it was sodium carbonate, sodium ions would be just bystander ions.But calcium ions form chelate with citrate 3:2, decreasing concentration of free hydrogen citrate + citrate ions.So the overall calcium carbonate effect is being base and chelating citrate.I.E., CALCIUM CHELATES WITH CITRATE, SO THIS WOULD TIE UP THE CITRATE. NOT WHAT IS DESIRED.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet86 Now back to this idea, trying to get rid of sodium in the picture:(Think about mixing magnesium sulfate + citric acid...)Health Benefits of Magnesium Citrate - WebMDMagnesium citrate is formed by combining magnesium oxide with citric acid, a reaction that createsmagnesium citrate and water. Magnesium citrate dissolves readily in water, so it may...Now, at this point, why not just take magnesium citrate straight and forget about making it with somethingelse.Look at this statement again:In addition, increases in free sodium load due to sodium citrate administration MAY increase intravascularblood volume, facilitating the excretion of bicarbonate compounds and an anti-urolithic effect."so urolithic is the production of stones, anti-urollthic will be the reduction of stone formation.NOTICE IT SAYS MAY. ALSO DO WE WISH TO EXCRETE BICARBONATE COMPOUNDS? Well, we know the boraxpolymer is certainly a carbonate compound, and we know that this is very much like a stone.Borax again is what?Borax is a sodium salt with boron. It is not a carbonate like baking soda or washing soda, which are. Youformed a carbonate precipitate by adding sodium carbonate to the decant. That definitely formed a carbonatecompound. You need to be careful here that you do enhance the formation of precipitates within the body.What are the risks of:too many sodium ions in the body?too many sulfate ions in the body?too many carbonates in the body?too much magnesium in the body?Lots of serious questions here.How much salt is too much?The U.S. Food and Drug Administration (FDA) recommends that adults eat less than 2,300 milligrams of sodiuma day. That!s about one teaspoon of table salt. #If you eat more than that in one day, it!s not going to hurtyou,$ says Zumpano. #But if you consistently exceed the recommended amount, you!re eating too much, andit can impact your health.$Most Americans consume about 3,400 milligrams of sodium a day. Only elite athletes or people who profuselysweat could possibly need this much sodium.soEffects of eating too much saltA sodium-heavy diet makes you retain fluid, which leads to swelling in the short term. But far more concerningis its impact on your body over time. Excess sodium intake can negatively affect your kidney function. It alsoleads to high blood pressure, the top risk factor for stroke.#If you only check your blood pressure once a year at your checkup, it could be normal that day,$ explainsZumpano. #Perhaps you!ve fasted, had a cup of coffee, peed a lot, and so your blood pressure seems fine. Butit might not be so normal if you were tested after eating out at a restaurant.$Signs your sodium intake is too highIt!s important to know how sodium affects you, as everyone is different. After eating a salty meal, look for thefollowing signs to see if you!re more sensitive to sodium!s effects.BloatingSodium attracts water. If you eat a lot of salty foods, you!ll experience fluid retention (when sodium holdswater in your body). The result? You feel swollen and look puffier, especially around the abdomen and eyes.You may also notice swelling in your hands and feet.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet87 Increased thirstRecent research shows higher levels of sodium don!t cause people to be thirstier. But despite that, somepeople can!t help but chug down more liquids after eating salty foods.Rise in blood pressureSodium is primarily located in our blood, and it pulls water to it. So, if you eat too much sodium, more waterenters your bloodstream. The greater volume of blood pushes against blood vessel walls, causing high bloodpressure (hypertension).Poor sleep qualityEating too much sodium can mess with your sleep, especially if you eat it in the evening. A spike in bloodpressure, along with urges to drink and pee, makes staying asleep difficult.How to flush sodium out of your systemIf you!re feeling the effects of too much salt, there are a few ways to rid yourself of excess sodium. "The verybest thing to do is sweat,# says Zumpano. "The body naturally removes sodium through sweat, tears andurine.#To even out your sodium level, get sweaty by exercising or sitting in a sauna. Drink plenty of fluids and eat alow-sodium diet the next day, too.Eating too much salt is bad for your health. Making a conscious effort to manage the sodium in your diet willpay off $ you!ll avoid the bloating and thirst that follows a salty meal and the risk of high blood pressure downthe road. That!s definitely a win-win.So we have a case where ONE foot is swelling at night. ONE FOOT. THE AFFLICTED LOWER, LEG AND ANKLE. Sothe indication is that we have a salt imbalance within THAT foot, but not both feet, so not general in any senseof the word. So the question is, what is actually producing the imbalance. Sodium attracts water. It says thatWITHIN that foot, there may be an excess of sodium. How and why is the question there?One thing we do know, the vascular system in the afflicted foot is totally screwed up. We also know from NIRthat the polymer withdraws the water from the skin. There is lots to think of here.This is really getting involved, difficult and interesting to sort out. Not all today....Look at my notes of Oct 23 2032 (handwritten notes) and we can see we did a series of ion tests on theculture. The most striking finding was the presence of calcium ions. We got a reading of 120 mg/L. This is high.Now we will study this article in the future closely, and watch closely what it says about calcium effects andlevels, and 120 mg/L is nothing to be dismissed.In the past five decades or so evidence has been accumulating about an environmental factor, which appearsto be influencing mortality, in particular, cardiovascular mortality, and this is the hardness of the drinkingwater. In addition, several epidemiological investigations have demonstrated the relation between risk forcardiovascular disease, growth retardation, reproductive failure, and other health problems and hardness ofdrinking water or its content of magnesium and calcium. In addition, the acidity of the water influences thereabsorption of calcium and magnesium in the renal tubule. Not only, calcium and magnesium, but otherconstituents also affect different health aspects. Thus, the present review attempts to explore the healtheffects of hard water and its constituents.Keywords: Alzheimer's disease, calcium, cancer, cardiovascular disease, diabetes, hard water, magnesium,reproductive healthGo to:INTRODUCTIONWater is essential for hydration and therefore, for life. It is also very important in food preparation and cooking,sanitation and hygiene, and a wide range of other uses. The drinking water supply has a primary objective ofprotecting human health, including ensuring access to adequate quantities of safe water. It is estimated thatapproximately 17% of the world's population uses water from the unprotected and remote sources, 32% fromsome form of protected sources and 51% from some sort of centralized (piped) system to the dwelling or aplot. Of the latter, a small but increasing proportion applies some form of treatment within the home.Individual water consumption occurs both at home and elsewhere, such as at schools and workplaces.Drinking-water is consumed not only as water per se but also in beverages and incorporated in food-stuffs. Inresponse to increasing global and local water scarcity, there is an increasing use of sources such asrecovered/recycled water, harvested rainwater, and desalinated water. 884 million people lack access to safeCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet88 water supplies; approximately one in eight people.[1] Among them a good percentage consumes hard water,which is considered to be a significant etiological factor around the globe causing many diseases such ascardiovascular problems, diabetes, reproductive failure, neural diseases, and renal dysfunction and so on.Hard water is usually defined as water, which contains a high concentration of calcium and magnesium ions.However, hardness can be caused by several other dissolved metals; those forms divalent or multivalentcations, including aluminum, barium, strontium, iron, zinc, and manganese. Normally, monovalent ions such assodium and potassium do not cause hardness. These divalent cations have a propensity to come together withanions in the water to form stable salts. The type of anion found in these salts distinguishes between the twotypes of hardness-carbonate and non-carbonate hardness [Table 1].Table 1Carbonate and non-carbonate hardness compoundsAn external file that holds a picture, illustration, etc.Object name is IJPVM-4-866-g001.jpgCarbonate hardness is caused by the metals combined with a form of alkalinity. Alkalinity is the capacity ofwater to neutralize acids and is attributed to compounds such as carbonate, bicarbonate, hydroxide, andsometimes borate, silicate, and phosphate. In contrast, non-carbonate hardness forms when metals combinewith anything other than alkalinity. Carbonate hardness is sometimes called temporary hardness because itcan be removed by boiling water. Non-carbonate hardness cannot be broken down by boiling the water, so it isalso known as permanent hardness. In general, it is necessary to distinguish between the two types ofhardness because the removal method differs for the two. Total hardness includes both temporary andpermanent hardness caused by the calcium and magnesium, on the basis of which water is categorized as softor hard and very hard [Table 2]. The ratio of calcium and magnesium in water is also a crucial factor indicatingthe hardness and in the causation of several hard water health problems. Hardness generally entersgroundwater as the water percolates through minerals containing calcium or magnesium. The most commonsources of hardness are limestone (which introduces calcium into the water) and dolomite (which introducesmagnesium.) Since, hardness enters the water in this manner groundwater generally has a greater hardnessthan surface water.[2]Table 2Concentrations of dissolved calcium and magnesium in soft and hard waterAn external file that holds a picture, illustration, etc.Object name is IJPVM-4-866-g002.jpgGo to:POTENTIAL HEALTH EFFECTSHard water has no known adverse health effect, WHO says at its Geneva Conference. In addition, hard water,particularly very hard water, could provide an important supplementary contribution to total calcium andmagnesium intake.[3] The health effects of hard water are mainly due to the effects of the salts dissolved in it,primarily calcium and magnesium. To a large extent, individuals are protected from excess intakes of calciumby a tightly regulated intestinal absorption mechanism through the action of 1, 25-dihydroxy-vitamin D, thehormonally active form of vitamin D. Although, calcium can interact with iron, zinc, magnesium, andphosphorus within the intestine, thereby reducing the absorption of these minerals. On the other hand, themajor cause of hypermagnesemia is renal insufficiency associated with a significantly decreased ability toexcrete magnesium.[4] Increased intake of magnesium salts may cause a change in bowel habits (diarrhea).Drinking-water in which both magnesium and sulfate are present in high concentrations (~250 mg/l each) canhave a laxative effect. Laxative effects have also been associated with excess intake of magnesium taken inthe form of supplements, but not with magnesium in the diet.Cardiovascular diseaseIn most large-scale studies, an inverse relationship between the hardness of drinking-water and cardiovasculardisease has been reported.[5,6,7,8,9,10,11] However, no such association has been found in some otherstudies,[12,13] particularly in those involving small geographical areas a clear association is often notfound.[14] The extent to which confounding variables, such as climatic, socioeconomic, or major risk factors,may account for the inverse relationship is unclear. Nevertheless, in a number of studies, a weak inverserelationship was reported after allowance was made for climatic and socioeconomic factors[15] and after majorrisk factors such as hypertension, smoking habits, and elevated serum lipids were taken into account.[16,17]An inverse relationship between hardness and cardiovascular disease has been reported in men after allowingfor climatic and certain social factors, but only up to about 170 mg of calcium carbonate/l.[18] A variety ofhypotheses have been proposed to explain the possible inverse association.[19,20,21,22,23,24] However,none has been fully substantiated, nor has a particular element been found to be conclusively associated withthe cardiovascular disease. It may be correlated with a high level of magnesium in hard water, which has someanti-stress actions against coronary heart disease. In a study of regional differences in mortality incardiovascular diseases in 76 municipalities in mid-Sweden a considerable gradient was found between thewestern areas with a high mortality and soft water and the eastern areas with a low mortality and hard water.The hardness of water defined as the sum of the content of calcium and magnesium, was shown to be ofCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet89 considerable influence of the differences in mortality compared to major risk factors.[16] The incidence ofcoronary heart disease varies widely in different geographical regions over the world and seriousepidemiological studies have been carried out to identify variables that could explain this fact. The role ofwater hardness has been widely investigated and evaluated for many years in several studies where regionaldifferences in cardiovascular disease have been discussed.[16,25,26,27,28,29] Earlier studies have foundpositive correlations between water and dietary magnesium and calcium and blood pressure.[30,31,32,33] InFinland and South Africa it was found that the incidence of death ascribed to ischemic heart disease isinversely correlated with the concentration of magnesium in drinking water[29] and also in a Swedish casecontrol study magnesium, and calcium in drinking water were associated with a lower mortality from acutemyocardial infarction in women but not with the total incidence.[26,34] However, other studies could notconfirm these findings,[31,32] conversely, in a study of magnesium in drinking water supplies and mortalityfrom acute myocardial infarction in North-West England, there was likewise evidence of an associationbetween magnesium and cardiovascular mortality.[25] In a Swedish study, the skeletal muscle magnesiumlevels were a significantly higher in persons living in an area with a higher water magnesium.[35] Theconcentration of magnesium in striated muscle has been used as a marker to evaluate the ion content in thesoft tissue.CancerImportant findings in this field were provided recently by Taiwanese scientists. In most of their studies, theauthors indicated a negative statistical association of various types of cancer morbidity/mortality with thehardness of water and calcium. In a review of these publications, it is worth noting the results concerning thepossible association between the risk of gastric cancer and the levels of calcium and magnesium.[36] Somestudies suggest there was a significant protective effect of calcium intake from drinking water on the risk ofgastric cancer. Magnesium also exerted a protective effect against gastric cancer, but only for the group withthe highest levels of magnesium exposure.[37] In another matched case-control study, the authors found apossible association between the risk of colon cancer and hardness levels in drinking water from municipalsupplies in Japan (obtained trend analyses showed an increasing odd ratio for the cancer with decreasinghardness in drinking water).[38,39] Similar epidemiological trends were also achieved for the relationsbetween hardness levels in drinking water, and the risk of rectal cancer and pancreatic cancer mortality,however, the researchers did not find any association with magnesium levels (the adjusted odds ratios werenot statistically significant for the relationship between magnesium concentrations in drinking water and rectalcancer).[40] One of the strongest epidemiological evidences of significant protective effect of magnesiumintake from the drinking water was that gave for the risk for esophageal cancer and ovarian cancer.[41,42]Unfortunately, these authors did not find any results pertaining to the similar trend between drinking-watermagnesium and liver cancer. The first strong evidence concerning the possible ecological relation betweenexposure to water magnesium and hepatic cancer was reported in Eastern Europe.[43]Cerebrovascular mortalitySome reports suggest there is a significant protective effect of magnesium intake from the drinking water onthe risk of cerebrovascular disease.[44] Despite their inherent limitations, studies on the ecological correlationbetween mortality and environmental exposures have been used widely to generate or discreditepidemiological hypotheses. Dietary calcium is the main source of calcium intake. Epidemiological studieshave shown that dietary calcium is inversely associated with the blood pressure. With much of theepidemiological literature suggesting a relationship between the dietary calcium and blood pressure, it wouldseem reasonable to expect that intake of dietary calcium could reduce the risk of cardiovascular events, suchas stroke that are commonly associated with hypertension.[45] However, controlling for magnesium levelseliminates the perceived effect of calcium levels on cerebrovascular mortality. In the general population, themajor proportion of magnesium intake is through food, and a smaller proportion is through drinking water. Forindividuals with the borderline magnesium deficiency, waterborne magnesium can make an importantcontribution to their total intake. In addition, the loss of magnesium from food is lower when the food is cookedin magnesium-rich water. Magnesium in water can also play a critical role because of its high bioavailability.Magnesium in water appears as hydrated ions, which are more easily absorbed than magnesium in food. Thecontribution of water magnesium among persons who drink water with a high magnesium levels could becrucial in the prevention of magnesium deficiency. The significant association between mortality from thecerebrovascular disease and the levels of magnesium in drinking water is supported by knowledge of thefunctions of magnesium. Magnesium is an enzyme (Na+/K+ ATPase) activator and regulates cellular energymetabolism, vascular tone, and the cell membrane ion transport. A lack of magnesium leads to a decrease inthe concentration of intracellular potassium and an increase in calcium levels. Magnesium deficiency mayincrease the contractility of blood vessels. Magnesium causes vasodilation by stimulation of endothelialprostacyclin release and in vivo, prevents vasoconstriction of the intracranial vessels after experimentalsubarachnoid hemorrhage. In addition, fear of cerebrovascular disease should not deter anyone from drinkingwater with a low magnesium levels. In conclusion, the results of the present study show that there is asignificant protective effect of magnesium intake from drinking water on the risk of cerebrovascular disease.This is an important finding for the Taiwan water industry and human health risk assessment.[46]Malformations of central nervous systemThere is good evidence that environmental influences must play some part, possibly a major part, in theetiology of neural tube malformations in the human embryo. Almost all that evidence, however, relates toCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet90 non-specific and uncertain markers of as yet unidentified specific teratogens. For example, the frequency ofmalformations of the central nervous system varies greatly from country to country.[47,48] It also varies fromarea to area within countries: In the United States for the period 1950-59, mortality from spina bifida(myelomeningocele) was 2-3 times greater on the Atlantic coast than on the Pacific coast;[49] in South Walesthe frequency of central nervous system malformations in the coal-mining valleys is almost twice as high as inthe coastal plain;[50] in England and Wales as a whole the frequency is highest in the north, the north-west,and Wales and lowest in the East, Southeast, and South.[51,52] It is higher in first-born than in later borninfants and in infants born to younger and older mothers than to mothers in mid-reproductive life.[51,53] It ishigher among infants born in the poorer than in the well-to-do strata of society.[54,55,56] It tends to be higheramong winter than among summer births.[51,53,55,57,58,59] Striking secular swings in frequency have beenreported in the Birmingham, Scotland, Dublin, and Boston. 70-71 Penrose[48] seems to have been the first tospeculate !the geographical variations observed in the incidence (of anencephalus) might suggest a# causalagent, such as the presence or absence of trace elements in the water supply." This suggestion has beentaken up by Fedrick.[60] She related data on anencephalus for 10 different areas in the United Kingdom (from10 different studies and relating to 10 different time-periods) to information about the water supplies of thoseareas obtained from various sources. Despite such manifestly unsatisfactory data, she found that thefrequency of anencephalus was a significantly related to measurements of the total hardness, calcium content,and pH of the local water supplies. Stocks[52] examined the mean annual death rates (still-births plus infantdeaths) for congenital malformations in the 15 hospital regions of England and Wales. Mortality rates werehighest in the north and the west and lowest in the Southeast. Having observed that mortality fromcardiovascular disease followed much the same regional pattern, he proceeded to correlate death rates fromcongenital malformations in the 15 regions with death rates of women aged 25-54 from the certain causes inthe corresponding regions. He found that mortality from malformations of the neural tube correlated veryclosely with mortality from the cardiovascular diseases, whereas other malformations produced insignificantnegative correlations. He concluded that because mortality from cardiovascular disease in the countyboroughs of England and Wales has been shown to be strongly an associated with softness of their watersupplies,[61] a water factor might be responsible for the regional variations of mortality from the centralnervous system malformations. In this paper, and against this background, we relate area differences inmortality from malformations of the central nervous system in South Wales to estimates of the hardness of thewater supplies in these areas. We also present new data on perinatal mortality from anencephalus in thecounty boroughs of England and Wales and relate them to estimates of the hardness of their water supplies.Alzheimer's diseaseThe issue of aluminum as a cause for Alzheimer's disease has been contentious. In special circumstances suchas renal failure and massive exposure to aluminum in certain occupations aluminum may cause brainpathology similar to Alzheimer Disease. However, there is no definite evidence of the role of this metal in thecausation or development of Alzheimer disease. In a survey of 88 county districts within England and Wales,rates of Alzheimer's disease in people under the age of 70 years were estimated from the records of thecomputerized tomographic (CT) scanning units that served these districts. Rates were adjusted to compensatefor differences in the distance from the nearest CT scanning unit and for differences in the size of thepopulation served by the units. Aluminum concentrations in water over the past 10 years were obtained fromwater authorities and water companies. The risk of Alzheimer's disease was 1.5 times higher in districts wherethe mean aluminum concentration exceeded 0.11 mg/l than in districts where concentrations were less than0.01 mg/l.[62]DiabetesHard water is indicative of the presence of higher levels of magnesium. In certain areas, drinking wateractually contains 100% or more of the recommended daily allowance about magnesium, which is around300-400 mg daily with levels varying according to gender and age. Because, all kinases and other ATP-relatedenzymes and channels regulating insulin action are dependent on magnesium, it is not surprising that serummagnesium concentrations have been found to be decreased in non-diabetic subjects with metabolicsyndrome and that hypomagnesaemia is a common feature in subjects with type-2 diabetes. Whether the lowintracellular magnesium content is secondary to or precedes insulin resistance is unclear; however, recentevidence suggests that sub-clinical magnesium deficiency may precipitate a diabetic state. Studies are neededto determine the role of sub-clinical magnesium status in diabetes risk. This should include measures ofglycosylated hemoglobin, an indicator of glycemic control that has been found to respond to oral magnesiumsupplementation and to correlate negatively with serum ionized magnesium or serum total magnesium in type2 diabetics.[63]Childhood atopic dermatitisAtopic dermatitis (or eczema) is an inflammatory, chronically relapsing, non-contagious and pruritic skindisorder. The environment plays an important part in the etiology of atopic eczema, but the specific causes areunknown. Exposure to hard water is thought to be a risk factor for eczema. The prevalence of symptoms ofatopic eczema among Japanese, Nottinghamshire and Spanish children is the most. The reasons for such ahigh prevalence are unknown. The study used data on water hardness and chlorine content of the watersupply; prevalence of atopic dermatitis diagnosed by physicians and episodes of wheezing reported by theparents; and potential confounding factors by socioeconomic and health-care status per municipality. Theprevalence of atopic eczema was significantly higher in the highest water hardness category than that in theCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet91 lowest respectively. A significant relationship between the chlorine content of the water supply and theprevalence of atopic dermatitis was observed after adjustment for confounding factors. Water hardness mayincrease the risk of atopic dermatitis among elementary-school children in Japan, as well as in the UnitedKingdom.[64]Kidney stonesThe hardness of water is due to the presence of carbonate and sulfate salts of calcium and magnesium. Morethan 3/4th of kidney stones are generally composed of calcium salt and usually occur as calcium oxalate andless commonly as calcium phosphate. The remaining 20% of stones are composed of uric acid, struvite andcystine stone. Stones form in urine that is supersaturated and this saturation is dependent on chemical free ionactivity, which makes the urine under-saturated. In this situation, the stone will not grow and may evendissolve. Increased urinary ion excretion and decreased urine volume will both an increase free ion activity andfavor stone formation and growth. Formation of kidney stones (nephrolithiasis) is based on genetic, metabolic,nutritional and environmental factors. Metabolic factors involved in stone formation include hypercalciuria,hypocitraturia (due to renal disease), hyperuricosuria, hyperoxalaturia, cystinuria and infections.Environmental and nutritional factors include dehydration, high salt intake, a diet rich in animal proteins andcalcium rich diet when oxalate intake is restricted. The impact of water hardness of urinary stone formationremains unclear, despite a weak correlation between water hardness and urinary calcium, magnesium, andcitrate excretion. Several studies have shown no association between water hardness and the incidence ofurinary stone formation. A correlation between water hardness and urinary calcium, citrate, and magnesiumlevels has been observed although the significance of this is not known. Some studies suggest that in thepreventive approach to calcium nephrolithiasis, intake of soft water has been preferable to hard water since itis associated with a lower risk for recurrence of calcium stones.[65]Reproductive healthThere are few reports of the effect of water hardness over reproductive health of men, most of thememphasized on the effect of its constituents, calcium, and magnesium,[66] while others on some otherconstituents like fluoride.[67] However, some reports show the occurrence of reproductive failure and stillbirthin India in hard water regions of India.[68,69] Some of these are showing the effect of excess calcium on thereproductive system and its negative influence on fertility.[68,70,71] These reports demonstrated theoxidative stress induced infertility in men by calcium,[72] but showed beneficial effects of magnesium.[4]There are also some reports of effects of waterborne fluoride on growth, reproduction and survival whichshowed long-term exposure of fluoride causes a progressive decline in reproduction.[67] On the contrary, infemale, magnesium sulfate present in hard water is indicated to prevent eclampsia in patients withpre-eclampsia. Magnesium sulfate decreases the risk of developing eclampsia around 50% and also decreasesmaternal mortality.[73] The WHO considers that magnesium sulfate is the elective drug for the prevention ofeclampsia in patients suffering from pre-eclampsia. Magnesium sulfate has also been demonstrated to preventpreterm labor.[74]Digestive health and constipationEven GI health is also reported being benefited from hard water since it provides potentially alleviating effectson the onset of constipation in the 85% cases. A rich union of calcium and magnesium in hard water, in a rightcombination, helps to combat constipation. The calcium in hard water results in teaming up with excess bileand its resident fats to lather up the soap like insoluble substance, which is emitted from the body duringbowel movements. Indeed, many renowned scientists have considered hard water as a boon as it has somefantastic health benefits that seem to encourage longer life expectancy and improved health. Magnesium saltrepresents with a laxative effect. This provides a rapid evacuation of intestine. Magnesium citrate, magnesiumphosphate, and magnesium hydroxide are also used. The American Gastroenterology Association recommendsmilk of magnesia for the management of constipation as one of the therapeutic options; however, theRehabilitation Nursing Foundation discourages the routine use of saline magnesium laxatives due to possibleside effects such as abdominal cramping, watery stools, and potential for dehydration and hypermagnesium.They only indicate the use of these laxatives in end-stage patients when other options have failed, and withand adequate prospective evaluation of magnesium levels.Bone mineral densityThe correlation between calcium and magnesium in drinking water and its impact on bone health areunidentified. There is some evidence that high-calcium water is beneficial to bone.[75] It has been reported ina study that spine mineral density was significantly higher in women aged 30-70 years living in Sangemini, aregion of central Italy, who drank the local high-calcium water (318 mg/l), compared with women in the sameregion who drank low calcium water (Other health effectsThe results of several studies have suggested that a variety of other diseases are also inversely correlated withthe hardness of water, including anencephaly[78,79] and various types of cancer.[80,81,82,83,84,85,86]However, the significance of these results is unclear, and it has been suggested that the associations mayreflect disease patterns that can be explained by social, climatological, and environmental factors, rather thanby the hardness of the water.Go to:CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet92 CONCLUSIONSHardness is important for drinking-water from the point of view of both aesthetic acceptability and operationalconsiderations. Although, there is some evidence from epidemiological studies for a protective effect ofmagnesium or hardness on cardiovascular mortality, the evidence is being debated and does not provecausality. Further studies are being conducted. In spite of this, drinking-water may be a source of calcium andmagnesium in the diet and could be important for those who are marginal for calcium and magnesium intake.Where drinking-water supplies are supplemented with or replaced by dematerialized water that requiresconditioning, consideration should be given to adding calcium and magnesium salts to achieve concentrationssimilar to those that the population received from the original supply. Consumers should be informed of themineral composition of their water when it has been altered by piped suppliers or treatment devicemanufacturers and by means for supplementing if desired. The contribution of drinking-water minerals formineral nutrition should be considered where changes in supply are proposed or where novel sources, such asseawater or brackish water, are exploited for drinking-water. There are insufficient data to suggest eitherminimum or maximum concentrations of minerals at this time, and so no guideline values are proposed.MOST DEFINITELY TO BE CONTINUED...Let's start looking at making probability estimates of the various functional groups now.Probability estimates are looking very reasonable and conservative at this point. 70% probablility level isprobably quite low for such an extensive data set. RMS looks to capture the data set at a much higher level, eg80, 85%.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet93 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.07 Functional Group Worksheet94 2023.12.08 Functional Group WorksheetContinuing:From Nov 16 2023 Notes:This exists as the most current interpretation of this material that has been created, a polymerized protein.Unclear at this point if the polymerization is likely to come from aromatic or amide origin, or both for thatmatter. We know that in the end a "rubberized" polymer is developed, but this can come from the other TSPsurface synth-cell polymer layer. But out of curiosity, is rubber aromatic or amide based?Rubber is alkene based. OK, this gets very interesting. Synthetic rubber is a copolymer between styrene andbutadiene. Styrene is a VINYL group attached to an aromatic ring. Butadiene is an alkene.Guess what, all the signs are starting to point the same direction. Aromatic, vinyl, alkenes are the componentsof synthetic rubber, which is most characteristic of the mature clot gross properties. Aromatic, vinyl, alkenesappear to be dominant structures of our NIR plots. Do not keep trying to force a single functional group. Thecombination of these three functional groups along with synthetic blood go a long way toward explaining theculture findings in general.So our three main components (everything is temperature and pH dependent for expression (~18 expressionsthus far) are:1. Synthetic blood (w/hemoglobin) (scores of proteins within blood) (surface layer of mature culture)2. A set of proteins that can be polymerized with borax, structurally firm but not elastic. (within decant)3. A vInyl contribution for elasticity of the culture. (within precipitate)This makes up the majority of the clot structure.1,3-Butadiene (a synthetic rubber) from NIST data base UV spectral search shows a peak at 293. This is a vinylcompound and then a match for our spectrum of TSP 100.6 at this point (not the rest of the spectrum nor our232 peak found.NIST and Chemspider both have some UV searching capability if you can get a part of the name. VERYVALUABLE!The styrene UV spectrum explains high absorbance towards the 232 peak. Synthetic rubber is a copolymerbetween styrene and butadiene.I must have looked up the UV on NIST for styrene based upon leads. Let's do this again.What are the physical properties of styrene? (AI)The physical properties of styrene include:1. Appearance: Styrene is a clear, colorless liquid.2. Odor: It has a sweet, floral, and somewhat fruity odor.3. Melting Point: The melting point of styrene is around -30.6 degrees Celsius (-23.1 degrees Fahrenheit).4. Boiling Point: Styrene has a boiling point of around 145 degrees Celsius (293 degrees Fahrenheit).5. Density: The density of styrene is approximately 0.91 grams per cubic centimeter.6. Solubility: Styrene is soluble in a wide range of organic solvents, such as ethanol, acetone, and benzene.7. Vapor Pressure: It has a relatively low vapor pressure at room temperature.8. Flammability: Styrene is highly flammable and can form explosive mixtures with air.9. Toxicity: Exposure to styrene vapor or liquid can cause irritation to the eyes, skin, and respiratory system.2023.12.08 Functional Group Worksheet95 Prolonged or high-level exposure may have more severe health effects.It is important to note that these properties may vary slightly depending on the specific grade or purity of thestyrene.what is the UV absorbance of styrene? (AI)The UV absorbance of styrene can vary depending on factors such as concentration, solvent, and wavelengthof light used for measurement. Generally, styrene exhibits weak absorption in the UV region. Some studieshave reported a maximum absorption peak around 250-260 nm. However, it is recommended to consultspecific literature or experimental data for accurate and detailed information on the UV absorbance of styreneunder specific conditions.This shows you that there is a lot of variability in the assessment of UV absorption of sytrene. Anywhere from232 (apparently? up to 260). That is a wide range. Profile of absorption is therefore important also.So where and how and why did I make the comment about styrene at 232 nm?Guess what, I can create my own spectrum to get me in the ball park with styrofoam. It is a form ofpolystyrene. Great.But we have this problem:"HCL will not melt the styrofoam. Needs to be petroleum based. For that matter, you could pour battery acid(sulfuric acid - H2SO4) in a styrofoam cup and it would not melt the cup. Both Hydrochloric and Sulfuric acidsare inorganic (they do not contain Carbon) Styrofoam does. . First rule in Chemistry-101, like dissolves like."This is important as it also affects interpretation of clot, synth-polymer layer, and the borax polymer. Need tonow revisit all of that.So acetone and gasoline work, but acetone may damage the quartz cuvettes? Let' see. At $20 a pop, I cannotafford that.So yes, acetone will damage quartz cuvettes. However, I can afford plastic cuvettes and this will get me downto 230 nm which should be adequate. So let's try that.Behavior of sample was strange so I reconstructed the control series, in two stages.1. 1st control is H2O.2. 2nd control is H2O + 100 ul acetone.3. Sample is H2O + 100 ul (acetone + sytrofoam mix) and then sample with H2O + 300 ul (acetone +styrofoam mix).Acetone has its own distinctive UV profile but that is to be dismissed and disregarded here. The only impact ofthe styrofoam was to shift the strong acetone peak at ~288 nm to:1. 290 nm with the 100 ul addition of the styrofoam.2. 298 nm with the 300 ul addition of the styrofoam.So there is nothing real dramatic here with, either way, no noticeably different peak behavior in the 230 - 260nm range. So these results are somewhat ambiguous.I find this information:Polystyrene (PSt) exhibits a new UV absorption band at around 290nm.https://link.springer.com/article/10.1007/BF00310794which is what we get for the lower concentration sample (slight shift from acetone). However, as we notice, astronger concentration shifted the peak further to the right. Nevertheless, from what we find and measuredirectlyThe UV peak of styrene looks to be about 244. Not exactly 232. Remember though that this is a borax polymer,it is solid and inflexible, so you do not expect it to be of rubber nature.So this says styrene is some where in the neighborhood of 240-260, depending on who you are listening to,and that polystyrene is somewhere on the order of 290 - 300 depending upon concentration,at least asdissolved in acetone.This is far too much variation for me and therefore attributing the peak at 232 to styrene seems useless andfruitless to me. You might try an NIR plot but I see no value in UV here.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.08 Functional Group Worksheet96 NOTE:The styrene UV spectrum explains high absorbance towards the 232 peak[NOTE DEC 08 2023 I SEE NO BASIS FOR THIS PREVIOUS STATEMENT - IT IS NOT ACCEPTED AT THIS TIME].Let's carry on keeping the styrene proposition in mind but not seeing any data collection for that compoundyet. I think we need a NIR spectrum.OK, NIR looks like a great way to approach this problem. Dissolving styrofoam in acetone created a residualpolymer material which did not remain in solution, It truly does form a visible plastic. Making an NIR card ofthis looks to be the best approach without any doubt. It is a good solid sample to work with, but in aputty-plastic form. The solution approach seems to be much harder to extract from. The NIR card should be agood test.I have an excellent NIR collection on styrofoam (polystyrene). Let's see what it shows.Polystyrene (Styrofoam) CI1146 (moderate) ArCH (Aromatic CH) (1143)(1142)1192 (weak) Methyl (1194) (1195)1405 (moderate) OH Methanol - OH with Hydrogen Bonding (1408) Methylene (1395)1638 (weak) CH from vinyl group (CH2=CH-) CH from vinyl associated group (1637) CH Vinyl - AliphaticHydrocarbons (1635)1682 (strong) Aromatic CH (1680) (1685) CH Methyl (1682)Certainly looks to be on track.Now look at notes of Nov 18. Instead of thinking only aromatic protein, with what you know about polystyreneand the dominant compoent of aromatic CH, you see that a progression of the following may well be in order:1. water soluble protein and vinyl groups (are there aromatics here as well?)2. non water soluble polyamide (like nylon, etc), not just consideration of an aromatic protein3. rubber clot (vinyl and styrene)."I have a very significant plot now to look at. It is a NIR comparison of TSP 100.5 (Decant) over a 12 dayincubation period. This is from the time that it is decanted (amber color) until it turns white approx 10 dayslater.Main feature I am looking at is that appears there is a bulk of mass (absorbance) shift from the Nov 06 TSP100.5 (amber) at a very broad peak of ~1620 +/- 20 nm to a more focused peak at Nov 18 TSP 100.5 (White)of 1516. They do however, continue to share a great deal, and in fact the majority of the turf from 1530 - 1650nm.Interpretation of this is:1620 is alkene and vinyl1520 is protein, 1515 is polyamidethis makes a lot of sense. It is saying it starts out stronger in the vinyl section but then shifts over to a proteinnature. However, it still retains the vinyl nature. This means that a conglomerate is being formed, i.e. apolymerized protein as your surmise.This is exactly what you envision happening. This answers a very important question on culture dynamics."What is really important here is the demonstrated transition or ships from alkene and vinyl (alcohol also?aromatic also?, tyrosine, etc) all water soluble towards a protein and polyamide, which we know to beinsoluble. This is a critical feature to demonstrate.IT IS POLYMERIZING.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.08 Functional Group Worksheet97 Now revisting the nucleic acid - DNA issue. This still remains as a distinct possibility. You now have two maturecultures to compare to one another TSP 070 and TSP 100. We see that as the culture matures, over 1-2months, the middle layer by UV presents a spectrum that seems to coincide quite strongly with nucleic acid -DNA properties. Even if we restrict ourselves to the 260/280 ratio we find:1. TSP 100: Nov 08 2023 (1 month age).77/.57 = 1.352. TSP 070 (Oct 08 2023 (2 months age)1.62 / 1.23 = 1.32Both samples were restricted to 300 ul in 3ml H2O.This presents an estimate of DNA (Nucleic acid) concentration of 15 % DNA, 85% Protein.Corrleation of profile with Nucleic Acids can also not be dismissed.This situation simply does not correspond with protein only. The profile as is cannot occur in a protein onlyenvironment and the peak for TSP 070 as a 60 day old culture is extremely well defined and far away from 280region. 280 region also is not peaking in any way as 260 area is. I say you have a strong case of nucleic acidsor DNA being present.Concentration graph (now modified) image source is:https://www.agilent.com/cs/library/applications/A260A280-spectral-scanning-5994-2538EN-agilent.pdfAlcohol test. NIR test.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.08 Functional Group Worksheet98 2023.12.09 Functional Groups DNAI now have the NIR of the middle layer of TSP 070 and TSP 100 for the nucleic acid inquiry. This will be itemizedhere but preliminary review shows that it supports the claim for nucleic acid/DNA existence.Also the following links are going to be helpful:https://bio.libretexts.org/Courses/University_of_California_Davis/BIS_2A%3A_Introductory_Biology_(Britt)/01%3A_Readings/1.17%3A_Protein_Structurehttps://socratic.org/questions/what-are-the-functional-groups-of-dnahttps://biologydictionary.net/hydroxyl-group/NIR TSP 100 (30 day culture) & TSP 070 (60 day culture). Both spectra are essentially identical, but TSP 070(older culture) is more clearly defined in general.1200 nm (moderate) OH (1200) hydroxyl1505 nm (strong) aromatic amine (1502.5) alkyl alcohol, containing one OH, R-C-OH (1500) NH combinationband (NH2-C=O - NH2 - urea)1597 nm (strong) polyamide (C=O/N-H combination) (1598)1642 nm (weak) vinyl group (CH2=CH-) (1637)1692 nm (moderate) Protein (CONH2) (1690) Aromatic CH (1689) Methyl (1693-1695) 8 instancesThis is an important discerning NIR plot. The first stage of analysis is to acquire the functional groups of bothproteins and DNA. I will discern from the NIR plot which of those exist and what can be associated with proteinor DNA. All functional groups will enter into the worksheet, however. The question confined here is whether ornot the DNA/nucleic acid prospect is supported above and beyond the UV 260/280 finding.The links now become important.The functional groups of DNA are:The functional groups are amine, amide, hydroxyl, glycoside linkage, and phosphodiesterThe functional groups of protein are:Amino acids are the monomers that make up proteins. Each amino acid has the same core structure, whichconsists of a central carbon atom, also known as the alpha (a) carbon, bonded to an amino group (NH2), acarboxyl group (COOH), and a hydrogen atom.Peptide bond formation is a condensation reaction. The carboxyl group of the first amino acid is linked to theamino group of the second incoming amino acid. In the process, a molecule of water is released and a peptidebond is formed. Try finding the backbone in the dipeptide formed from this reaction. The pattern you arelooking for is: N-C-C-N-C-CThe sequence and the number of amino acids ultimately determine the protein's shape, size, and function.Each amino acid is attached to another amino acid by a covalent bond, known as a peptide bond, which isformed by a dehydration synthesis (= condensation) reaction. The carboxyl group of one amino acid and theamino group of the incoming amino acid combine, releasing a molecule of water and creating the peptidebond.and this is a crucial statement:Amino acids are the function units of protein. Each amino acid molecule contains a carboxyl group at one end,and an amino group at the other end. Amino groups consist of a nitrogen bonded to two hydrogens, which isattached to the carbon backbone of the amino acid. When the protein is formed, the carboxyl group loses thehydroxyl group attached to it, while the amino group loses a hydrogen. With the loss of these molecules, theamino group binds to the carbonyl group, forming a peptide bond. What else is produced during this reaction?So after the protein is formed, the hydroxyl group no longer exists, and water is formed in the process. Oursample has been dried, however, and all water removed. Therefore there should be no hydroxyl group presentin our NIR sample.2023.12.09 Functional Groups DNA99 However, DNA maintains a hydroxyl group. Our NIR plot is therefore further supportive of the DNA prospectwithin the TSP cultures, above and beyond the UV which shows a strong 260/280 ratio presence. This is acritical assessment.The fully chilled ethanol DNA tests (both sample and ethanol are chilled prior to mixing) are fully successful inindicating the presence of DNA. There is a white layer formed at the alcohol-sample interface. Materialproduced is minimal but the appearance is fully in keeping with all previous DNA control tests over the years.All three DNA inquiry tests are with positive results:1. UV 260/280 absorbance ratio.2. NIR Hydroxyl group detection3. Chilled alcohol tests.This is an important achievement. Bear in mine that DNA extraction was succussful on 3 prior events in pastyears, so this is once again fully consistent. What is different here is that the method derives from a watersoluble portion of a mature culture. No solid extraction processes were required.Now we return to the worksheet.These are the NIR peaks of TSP 100.5 identified on Nov 18 2023: (CONCENTRATED Decanted TSP 100.0 postTitration)TSP 100.5 NOV 18: 1397 1520 1598 16461397 Methyl (1396) Methylene (1395)1520 Amide/protein (1520) Amine (1520)1598 Polyamide (C=0/N-H) (1598)1646 Vinyl (1637) Methyl (1654)The acid-base behaviors of the:1. synth polymer2. carbonate precipitate3. rubberized blood clotneed to be sorted out.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.09 Functional Groups DNA100 2023.12.10 Functional Group WorksheetContinuing, now with Nov 23 2023.The acid-base behaviors of the:1. synth polymer2. carbonate precipitate3. rubberized blood clotneed to be sorted out.Human blood-synth layer comparison to continue.Bacteriophage issue has not been addressed properly.Mid IR exploration of vinyl group with historical "CDB Protein Complex" IR data (cerca 2015) looks to be mostimportant. Initial peak match search brings up following candidates as:1. Butyl vinyl ether C6H12O2. Cyclohexyl vinyl ether C6H14OThese candidates have essentially exact matches with IR collected Sep 2015 of "CDB Protein Complex".Out of about 10 candidates from entire SDBS database. These are lowest molecular weight. I am guessing wehave something here.Look at physical properties, synthetic rubber, etc.Combination of human-synth blood comparison with polymerization probable is a worth topic.A new variation on the growth forms. The fungal filament form is well established. I do have a major culture ofonly that form that has been developed over the last 2-3 months. It creates a large mass ofmold--fungal-filament appearing material.What is different today is that a spare portion of TSP 100 set aside is producing a pure large growth form on itssurface, approx 2" x 1". A large central mass with appendages extending out on the surface. Colors are grey,white, pink and blue grey. It is a very clean form of this variation that comes fom a purified TSP 100 culture of~30 days age. This was overflow material from the culture that has been sitting idle. This culture produces thesynth-polymer layer also. It will be of interest if the synth cells can be identified within such a mass, as if so, itrepresents a clotting form in itself. It is solid. Database image included.This is a major topic that looks like it was never followed up with, notes are from Nov 27 2023:I am now in a position to start comparing the NIR of a blood clot to the NIR of the synth polymer layer. Themain uncertainties of comparison are:1. There is some ambiguity whether the clot comes from the deceased individual or a live individual (externallyclotted). The bias is strongly towards the deceased clot which is favorable, however this will likely be repeated.2. The trend is removed from the blood clot but not from the synth-polymer layer so no magnitudes of any kindcan be considered.With that being said, here is what we start to see. The main issue here involves similarity or difference in basicstructure and solubility. One thing we do know is that the clot dissolves in NaOH and the synth polymer layerdissolves in strong H2SO4. So we know already they are very different from one another and yet thepresumption is that they originate generally from the same source material. This means, since we knowe theyhave important differences, that comparison of the clot with other forms observed will be made.NIR Comparison:Start with the blood clot:2023.12.10 Functional Group Worksheet101 Clot: 1130 (1130) 1374 (1370) 1443 (1440) `1575 (1570, 1580)and then the synth-polymer layer:Synth-Polymer: 1195 (1190, 1200) 1520 (1500) 1594 (1590) Broad 1582-1609 (1580-1610) 1700 (1700)Clot:1130 (1130) ArCH (1142, 1143)1374 (1370) Methyl (1370)1443 `Aromatic Amine (1443) (1445) (1441) (1446) OH from sugar (1441) Methylene (1440) ArCH (1446)1575 (1570, 1580) Amide (1570) Polyamide (1570) Alcohol (1580) OH (1580)We are now at a very important juncture point in the work, and this is the NIR analysis of the blood clots thattook place approximately in Jun of 2023. First step is to recall the notebook volume(s). It appears to be Vol 30.Jun 22 2023 mentions receipt of blood clots.The first plot visited is that of the Live Individual, Vaxxed. Plot is detrended, normalized. All of the abovematches fine, but two more peaks are to be added:1504 Aromatic Amine (1502.5)1626 Vinyl, Vinyllidene (1630) (1621) (1631)I see now that the NIR of the blood clots must be revisited. I do not have all data that could be available. Firstissue is that there is some confusion on which sample is being used above. All direct statements state that it isSample C, which is from a living individual whose blood was allowed to settle outside the body and formed aclot. My notes (i.e., NIR plot) sometimes refer to it as from the deceased individual and this is where itbecomes confusing. I do think it is from Sample C notes of Jun 22 2023.We have enough information to establish the overlap of the culture with the blood clot. But let's look at theclots again and attempt to prepare NIR cards for each sample that we have.I have now prepared NIR cards of all three clot samples, L, C, D.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.10 Functional Group Worksheet102 CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.10 Functional Group Worksheet103 2023.12.11 Functional Group WorksheetLots of data now available to interpret. chi-square application?I have vinyl gloves here - solubility tests.Are DNA results included in worksheet?Variability of clot NIR, additional clots.Protein, polymer, synth blood, filaments are all in place.I have a strong NIR plot of the deceased clot. One question I have is variability between other clots howeverthe deceased clot seems to be the epitome of the end of the progression under study. Rubber nature is strongwith this clot.Blood Clot Deceased:935 Methylene (930) (930) weak984 ArOH (990) Water (979) Not Likely, sample is dried. strong1179 Alkenes, polyenes (1170) weak1422 ArOH (1420) Phenolic OH (1420) Aromatic CH (1417) strong1454 Aromatic Amine (1452.5) Water (1453) Not LIkely, sample is dried Alcohol (1452) weak1518 Amide/Protein (1520) Amine (1520) Polyamide (1515) strong1648 Methyl (1654) Vinyl (1637) strong1682 Methyl (1682) Aromatic CH (1685) moderateThis certainly looks like a very representative sampling of the functional groups encountered.OK, the analyis of clot vs., culture progression is complete.ON TAP:1. Finish paper.2. Bacteriophage inquiry.3. Chi square investigations.4. Binomial test?The paper is proceeding well and falling into place at the level required. This will give me some breathingroom.CI BLOOD CLOT PROGRESSION WORKSHEET (not original formatting)Native Form Native Form Native FormDate of Methods of Date Origin Age In H2O H2O pH HYDROCARBONS ALCOHOLS SDBS PROTEIN POLYMERStudy Sample Name Study of Sample Days Soluble Insoluble (0 if Unknown) Compound 1 Compound 2Compound 3 Compound 4 Compound 5 Compound 6 Compound 7 Compound 8 Compound 9 Compound 10Compound 11 Compound 12 Compound 13 Compound 14 Compound 15 Compound 16 Compound 17Compound 18 Compound 19 Compound 20 Compound 21 Compound 22 Compound 23 Compound 24Compound 25 Compound 26 Compound 27 Compound 28 Compound 29 Compound 30 Compound 31Compound 32 COMMENTS10/28/23 TSP 070 Decant LC, UV, NIR 10/08/23 20 1 acidic Alkyl Alcohol Ether Alcohol Ester Alcohol ProteinTyrosine Calcium Ions Nanoparticles10/29/23 TSP 070 Decant LC, UV, NIR 10/08/23 21 1 acidic Alcohol Protein Tyrosine2023.12.11 Functional Group Worksheet104 10/29/23 TSP Carbonate Decant LC, UV, NIR 10/08/23 21 1 acidic Methyl Alcohol Alkyl Alcohol Protein AmideAromatic CH Aromatic AmineMethyl Alcohol Aromatic Amine10/30/23 2nd Level Decant from Carbonate Decant UV 10/08/23 22 1 12.0 Protein10/30/23 2nd Level Precipitate from Carbonate Decant - H2SO4 Hydrolyzed UV 10/08/23 22 1 acidic Protein10/31/23 Human Blood NIR 10/31/23 0 1 7.4 est Vinyl VinylideneVinyl Vinylidene10/31/23 Synthetic Blood - Polymer NIR 10/08/23 23 1 acidic Methyl Alcohol Amine Polyamide Aromatic AminePolyamide11/02/23 Synthetic Blood - Polymer Meter 10/08/23 25 1 acidic Hemoglobin MeterKastle Meyer Hemoglobin Kastle Meyer11/03/23 Human Blood - REFERENCE MID IR 08/16/15 3001 1 7.4 est Ethyl Ether Vinyl High level confirmationof ethyl vinyl ether (or related compounds) in the blood of 2015 and 2017 with MID IR11/04/23 Synthetic Blood - Polymer - (H2SO4 Hydrolyzed) VIS-NIR 10/08/23 27 1 solid Methyl Alcohol AlkylAlcohol Aromatic CHMethyl Aromatic CH11/05/23 Synthetic Blood - Polymer - (H2SO4 Hydrolyzed) DNA PROSPECT UV 10/08/23 28 1 solid Nucleic Acids- DNANucleic Acids - DNA11/07/23 TSP 100.5D (M) 7.20-1110 CONCENTRATED Decanted TSP 100.0 post Titration NIR 11/06/23 1 1acidic Alkenes Vinyl11/07/23 TSP 70.1 Titrate Decant UV 10/08/23 30 1 acidic Alkyl Alcohol Bacteriophage?11/07/23 TSP 70.5 Centrifuged TSP 70.0 Bottom Layer NIR 10/08/23 30 1 solid Methyl Alkyl Alcohol ProteinAmide Aromatic Amine11/07/23 TSP 70.2 Centrifuged Titrate Precipitate NIR 10/08/23 30 1 acidic Methylene Polyamide VinylVinylidene AcrylatePolyamide VinylVinylVinylVinyl11/13/23 TSP 100.6C (B) X.XX-1110 Centrifuged Precipitate TSP 100.0 post TitrationNIR 11/10/23 3 1 acidic Methyl Alkyl Alcohol Vinyl Two layers grey/white centrifuge precipitate, grey dominates11/13/23 TSP 100.5D (M) 7.20-1110 CONCENTRATED Decanted TSP 100.0 post Titration UV, VIS-NIR 11/10/23 31 acidic Protein Tyrosine Amber solution 95% of volume11/13/23 TSP 100.0N (M) 3.81-1108 VIS-NIR 11/08/23 5 1 acidic Alkyl Alcohol Protein11/13/23 Urine - Polymer (Blood) Matrix Vinyl (Polymer Deduction)11/13/23 TSP 100.5D (M) 7.20-1110 CONCENTRATED Decanted Polymer Suface Layer Microscope 11/10/23 3 1solid Protein Tyrosine Vinyl (Observation 250nm) Amber solution 95% of volume11/15/23 Borax Polymer (developed in lab from TSP 100.5 & Borax) NIR 11/10/23 5 1 solid Methyl MethyleneAlkenes Phenolic OH Protein Amine Amide Polyamide Aromatic CH Butadiene (293 nm)Polyenes11/18/23 TSP 100.5D (M) 7.20-1110 CONCENTRATED Decanted TSP 100.0 post Titration 11/10/23 8 1 acidicMethyl Protein Amine Amide Polyamide Vinyl11/20/23 TSP 070 UV 10/08/23 43 1 acidic Nucleic Acids - DNA11/20/23 TSP 100 UV 11/08/23 12 1 acidic Protein Tyrosine11/21/23 TSP 070 UV 10/08/23 44 1 acidic Nucleic Acids - DNA11/22/23 Synthetic Blood - Polymer NIR 10/08/23 45 1 solid Amine Amide Polyamide Vinyl11/23/23 CDB Protein Complex 2015 MID IR 06/01/15 3097 1 film Butyl Ethyl Ether (SDBS)Ethyl Vinyl Ether (SDBS)Cyclohexyl vinyl ether C6H14O (SDBS)11/23/23 Synthetic Blood - Polymer - (H2SO4 Hydrolyzed) TSP 070 UV 10/08/23 46 1 solid Protein AmideNucleic Acids - DNA11/23/23 Synthetic Blood - Polymer - TSP 070 - Dried. Combined with H2SO4 Qualitative 10/08/23 46 1 solidHemoglobin11/09/23 TSP 070 Native pI Determination - Match Human Blood Proteome Titration 10/08/23 32 1 acidicHemoglobin11/24/23 Synthetic Blood - Polymer - (H2SO4 Hydrolyzed) - Purple - Sulfhemoglobinemia NIR 10/08/23 47 1solid Methylene Phenolic OH Hemoglobin Aromatic OH11/26/23 Human Blood - Vinyl Inquiry NIR 11/07/15 2941 1 7.4 est VinylVinylVinylVinylVinylideneVinylideneAcrylate11/26/2023 Human Blood - Vinyl Inquiry MID IR 11/07/15 2941 1 7.4 est Ethyl Vinyl Ether (SDBS) Ether Vinyl12/08/2023 TSP 070 DNA UV 10/08/23 61 1 Nucleic Acids - DNANIR 10/08/23 Nucleic Acids - DNACI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.11 Functional Group Worksheet105 Qualitative 10/08/23 Nucleic Acids - DNA12/08/2023 TSP 100 DNA UV 11/08/23 30 1 Nucleic Acids - DNA12/10/2023 Blood Clot Live Vaxxed NIR 06/23/2023 170 1 solid Methyl Methylene Alcohol Amide PolyamideAromatic CH Aromatic Amine Vinyl Vinylidenesolid Alcohol Aromatic CH Aromatic Amine Vinyl VinylideneAlcohol Aromatic Amine Vinyl VinylideneAromatic AmineAromatic Amine12/11/2023 Blood Clot Deceased NIR 06/23/2023 171 1 solid Methyl Methylene Alkenes Polyenes AlcoholProtein Amine Amide Polyamide Aromatic CH Aromatic OH Aromatic Amine VinylMethyl Aromatic CH Aromatic OHCount 36 20 16 RMS 7.1 12 1 5 3 2 9 7 2 1 1 4 2 14 5 5 5 8 9 8 3 10 9 1 23 8 2 1 0 0 0 1 1Range 23.0cFREQUENCY 0.552490 32 78 65 53 87 84 53 32 32 73 53 92 78 78 78 86 87 86 65 89 87 32 95 86 53 32 0 0 0 32 32Combined Clot AllGroups Frequency Pr% Groups Method MethodsVinyl 95 X NIR UVMethyl 90 X NIR NIRProtein 90 X NIR LCAromatic Amine 90 X NIR MID IRAlcohol 85 X NIR VISAlkyl Alcohol 85 NIR MicroscopyAmide 85 X NIR TitrationPolyamide 85 X NIR QualitativeAromatic CH 85 X NIR MeterNucleic Acids - DNA 85Vinylidene 85 NIRMethylene 80 X NIRHemoglobin 80Tyrosine 80Amine 80 X NIRAlknesN=15/30 Polyenes 10/12 36 samplesAromatic OHCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.11 Functional Group Worksheet106 2023.12.12 Research Paper - RegroupRegrouping.1. Research paper needs to pull itself in toward close. Look at bring Prob level to 75% vs 80%. How does itaffect inclusion/overlap list?It does not change anything. Leave at 80% Probability.2. Does vinyl dissolve in acid/base? Compare to clot behavior.Vinyl does not dissolve quickly or easily in either strong H2SO4 or NaOH-KOH.Same as clot behavior. This is therefore, an important observation.3. Outline the chemistry/functional groups towards synthetic rubber.From previous notes:"Rubber is alkene based. OK, this gets very interesting. Synthetic rubber is a copolymer between styrene andbutadiene. Styrene is a VINYL group attached to an aromatic ring. Butadiene is an alkene.Guess what, all the signs are starting to point the same direction. Aromatic, vinyl, alkenes are the componentsof synthetic rubber, which is most characteristic of the mature clot gross properties. Aromatic, vinyl, alkenesappear to be dominant structures of our NIR plots. Do not keep trying to force a single functional group. Thecombination of these three functional groups along with synthetic blood go a long way toward explaining theculture findings in general.2. Two polymeric forms, one for structual density and the other for elasticity. Both highly insoluble, even inmany strong reagents/acids/caustics.This is a major achievement. It would certainly be of benefit to understand how the two polymers may jointogether.Now the rubber property would be imparted by the vinyl group, which we now understand is a part of thecopolymeric process of synthetic rubber. We may well have, and are expected to have a copolymerarrangement here.The picture looks increasingly clear. It would seem that we have a genetically engineered co-polymerizationprocess taking place here. The process involves synthetic blood, an aromatic protein polymer, and a syntheticrubber equivalent polymer. Their origin is the Cross Domain Bacteria (CDB). The result is increased coagulationor a clotting of the blood. The end result is definitely a lethal threat.Mid IR exploration of vinyl group with historical "CDB Protein Complex" IR data (cerca 2015) looks to be mostimportant. Initial peak match search brings up following candidates as:1. Butyl vinyl ether C6H12O2. Cyclohexyl vinyl ether C6H14OThese candidates have essentially exact matches with IR collected Sep 2015 of "CDB Protein Complex".Out of about 10 candidates from entire SDBS database. These are lowest molecular weight. I am guessing wehave something here.Look at physical properties, synthetic rubber, etc.now tell me three common ways that synthetic rubber is made.Sure! Here are three common ways that synthetic rubber is made:1. Polymerization: This method involves the polymerization of monomers to form long chains of rubbermolecules. The most common monomers used in synthetic rubber production are butadiene and styrene.Polymerization can be achieved through various techniques such as emulsion polymerization, solutionpolymerization, or bulk polymerization.2. Copolymerization: In this approach, two or more different monomers are combined to create copolymers. Byblending different monomers, the resulting synthetic rubber can have enhanced properties compared torubber produced from a single monomer. For example, the copolymerization of butadiene and styreneproduces a type of synthetic rubber called Styrene-Butadiene Rubber (SBR), which is widely used in tiremanufacturing.2023.12.12 Research Paper - Regroup107 3. Vulcanization: Vulcanization is a process used to improve the properties of synthetic rubber. It involves theaddition of various chemicals, such as sulfur, accelerators, and activators, to the rubber compound. The rubberis then heated, which causes the chemical additives to react and cross-link the rubber molecules. Thiscross-linking process enhances the rubber's strength, elasticity, and resistance to heat, aging, and abrasion.These are just a few common methods used in the production of synthetic rubber. The specific techniquechosen depends on the desired properties and applications of the final rubber product.is butadience a vinyl product?No, butadiene is not a vinyl product. Butadiene is a colorless gas that is used as a raw material in theproduction of synthetic rubbers, such as styrene-butadiene rubber (SBR) and polybutadiene rubber (PBR). Vinylproducts, on the other hand, are made from vinyl chloride monomer and include materials like polyvinylchloride (PVC).what are examples of polymer vinyls?Some examples of polymer vinyls include:1. Polyvinyl chloride (PVC): PVC is a widely used polymer vinyl that is known for its versatility and durability. Itis commonly used in construction materials, pipes, electrical cables, and vinyl flooring.2. Polyvinyl acetate (PVAc): PVAc is a polymer vinyl often used in adhesives, paints, and coatings. It is also themain component in white glue or wood glue.3. Polyvinyl alcohol (PVA): PVA is a water-soluble polymer vinyl that is used in various applications such asadhesives, coatings, and textiles. It is also used in the production of PVA films and fibers.4. Polyvinylidene chloride (PVDC): PVDC is a polymer vinyl that is known for its excellent barrier propertiesagainst gases and moisture. It is commonly used in food packaging films, shrink wrap, and protective coatings.5. Polyvinylpyrrolidone (PVP): PVP is a polymer vinyl that is used in pharmaceuticals, cosmetics, and personalcare products. It is often used as a binder, film-former, or stabilizer in various formulations.These are just a few examples of polymer vinyls, and there are many more variations and derivatives availablein the market.what are the chemical and physical property differences between dienes and trienes?Dienes and trienes are both types of hydrocarbons that contain double bonds. However, there are somedifferences in their chemical and physical properties:1. Chemical Properties:- Dienes have two carbon-carbon double bonds, while trienes have three carbon-carbon double bonds.- Dienes can undergo addition reactions with electrophiles, such as hydrogenation or halogenation, at eachdouble bond independently. Trienes can also undergo similar addition reactions, but with the presence of threedouble bonds, the reactions can be more complex.- Dienes can undergo polymerization reactions to form polymers, such as synthetic rubbers. Trienes can alsoundergo polymerization, but the presence of more double bonds can lead to more complex polymer structures.- Dienes can undergo Diels-Alder reactions, which involve cycloaddition reactions with dienophiles. Trienes canalso participate in Diels-Alder reactions, but with the presence of more double bonds, the reactions can havemore diverse outcomes.2. Physical Properties:- Generally, as the number of double bonds increases from dienes to trienes, the boiling points and meltingpoints tend to increase. This is due to the increased intermolecular forces (Van der Waals forces) between themolecules.- The presence of double bonds in both dienes and trienes can lead to increased reactivity, making them moreprone to oxidation and other chemical reactions compared to saturated hydrocarbons.- Both dienes and trienes are typically liquids or low-melting solids at room temperature, depending on thespecific structure and molecular weight.It's important to note that these properties can vary depending on the specific diene or triene compound, asdifferent substituents and structural arrangements can influence their behavior.so we know that dienes and trienes are alkanes.CI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.12 Research Paper - Regroup108 what it is called when you have two vinyl functional groups combinedWhen two vinyl functional groups are combined, it is called a vinyl group dimerization or vinyl dimerization.This makes up the majority of the clot structure.1,3-Butadiene (a synthetic rubber) from NIST data base UV spectral search shows a peak at 293. This is a vinylcompound and then a match for our spectrum of TSP 100.6 at this point (not the rest of the spectrum nor our232 peak found.NIST and Chemspider both have some UV searching capability if you can get a part of the name. VERYVALUABLE!The styrene UV spectrum explains high absorbance towards the 232 peak. Synthetic rubber is a copolymerbetween styrene and butadiene.Chemicals that dissolve rubberTo understand what dissolves rubber, we need to examine the chemical composition of rubber itself. Rubber isprimarily composed of polymers, which are long chains of repeating units called monomers. While rubber isresistant to many chemicals, there are specific substances that can break down these polymer chains anddissolve rubber. Some of these chemicals include:Organic solvents: Certain organic solvents, such as acetone and benzene, have the ability to dissolve rubber.These solvents can penetrate the polymer chains and weaken the structure of rubber, causing it to breakdown.Strong acids and bases: Strong acids, like sulfuric acid, and strong bases, like sodium hydroxide, can alsodissolve rubber. These chemicals react with the rubber"s polymer chains, causing them to break apart anddissolve.Certain oils and fuels: Some oils and fuels, particularly those with high aromatic content, can dissolve rubber.These substances can penetrate the rubber and cause it to swell, leading to its eventual dissolution.Factors affecting rubber dissolutionSeveral factors can influence the dissolution of rubber. These include:Temperature: Higher temperatures can increase the solubility of rubber in certain chemicals. Therefore,exposure to elevated temperatures can accelerate the dissolution process.Duration of exposure: The longer rubber is exposed to dissolving agents, the greater the chances of it beingdissolved. Prolonged exposure allows the chemicals to penetrate deeper into the rubber, leading to itsbreakdown.Rubber composition: Different types of rubber have varying resistance to specific chemicals. Factors such asthe presence of additives or the type of polymer used in the rubber"s composition can affect its susceptibilityto dissolution.Tell me about synthetic rubber.Synthetic rubber is a type of rubber that is artificially produced rather than being derived from natural sourcessuch as latex from rubber trees. It is created through a chemical process known as polymerization, wherevarious monomers are combined to form long chains called polymers.There are several types of synthetic rubber, each with its own unique properties and uses. Some commontypes include:1. Styrene-butadiene rubber (SBR): SBR is one of the most widely used synthetic rubbers. It is highly resistantCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.12 Research Paper - Regroup109 to abrasion and has good flexibility, making it suitable for applications such as tires, conveyor belts, andfootwear.2. Polybutadiene rubber (BR): BR is known for its exceptional elasticity and low-temperature flexibility. It iscommonly used in the production of tires, as well as in various industrial applications where high resilience isrequired.3. Neoprene: Neoprene is a versatile synthetic rubber that is resistant to oil, heat, and weathering. It iscommonly used in the manufacturing of wetsuits, gaskets, hoses, and electrical insulation.4. Nitrile rubber (NBR): NBR is known for its excellent resistance to oil, fuel, and chemicals. It is widely used inthe automotive and aerospace industries for applications such as seals, gaskets, and hoses.5. Ethylene-propylene-diene rubber (EPDM): EPDM is a synthetic rubber with excellent weather resistance andelectrical insulation properties. It is commonly used in roofing materials, automotive parts, and electricalinsulation.Synthetic rubber offers several advantages over natural rubber, including better resistance to wear, aging, andchemicals. It can be formulated to have specific properties, making it suitable for a wide range of applications.Additionally, synthetic rubber production helps reduce dependence on natural rubber sources and providesmore consistent supply and quality control.However, synthetic rubber production also has environmental implications. The process of manufacturingsynthetic rubber involves the use of petrochemicals, which are derived from non-renewable fossil fuels. Effortsare being made to develop more sustainable and eco-friendly alternatives to synthetic rubber production.Is vinyl used in synthetic rubberNo, vinyl is not used in the production of synthetic rubber. Synthetic rubber is typically made frompetroleum-based materials such as styrene and butadiene. Vinyl, on the other hand, is a type of plastic madefrom vinyl chloride monomers.what is butadieneButadiene is a colorless gas with a mild aromatic odor. It is a highly flammable compound that is commonlyused in the production of synthetic rubber. Butadiene is also used as a chemical intermediate in themanufacturing of various products, including plastics, resins, and coatings. It can be produced from petroleumor by the thermal cracking of natural gas liquids.Does butadiene have any vinyl groups within itNo, butadiene does not have any vinyl groups within it. Butadiene is a hydrocarbon compound with thechemical formula C4H6, consisting of four carbon atoms and six hydrogen atoms. It is a conjugated diene,meaning it has two double bonds between carbon atoms.What is a vinyl functional groupA vinyl functional group is a chemical group consisting of two carbon atoms bonded together with a doublebond. It is represented by the symbol "-CH=CH2". The vinyl group is commonly found in organic compoundsand is known for its reactivity in various chemical reactions. It is often used in the synthesis of polymers, suchas polyvinyl chloride (PVC), and in the production of pharmaceuticals and other organic compounds.What is styreneStyrene is a colorless liquid hydrocarbon that is derived from petroleum. It is primarily used in the productionof polystyrene, a versatile plastic material commonly found in packaging, insulation, and disposable products.Styrene is also used in the manufacturing of other plastics, rubber, resins, and synthetic fibers. It is importantto note that prolonged exposure to styrene can have health effects, and it is classified as a possible humancarcinogen by the International Agency for Research on Cancer.Is there a relationship between a diene and a vinyl groupCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.12 Research Paper - Regroup110 Yes, there is a relationship between a diene and a vinyl group. A diene is a molecule that contains two doublebonds, while a vinyl group is a functional group consisting of a carbon-carbon double bond. In a diene, one ofthe double bonds can be considered as a vinyl group.THIS IS CRUCIAL INFO. YOU HAD TO KNOW WHAT TO ASK TO GET THS INFO. OTHERWISE VINYL RELATIONSHIPWAS NOT DISCLOSED.How do you make synthetic rubber from butadiene and styreneTo make synthetic rubber from butadiene and styrene, you can follow these steps:1. Obtain butadiene and styrene: Butadiene and styrene are both commercially available chemicals. You canpurchase them from chemical suppliers.2. Polymerization: Mix the butadiene and styrene in the desired ratio. Typically, a ratio of 75-80% butadieneand 20-25% styrene is used for making a synthetic rubber called styrene-butadiene rubber (SBR).3. Initiation: Add a suitable initiator, such as a peroxide or an organic metal compound, to the mixture. Thisinitiates the polymerization reaction.4. Polymerization reaction: Heat the mixture to a specific temperature, typically around 50-70 degrees Celsius,to initiate the polymerization reaction. The reaction can be carried out in a reactor vessel under controlledconditions.5. Stirring and heating: Stir the mixture continuously to ensure uniform mixing and heat it for a specificduration to complete the polymerization process. The reaction time can vary depending on the desiredproperties of the synthetic rubber.6. Cooling and solidification: Once the polymerization is complete, cool the mixture to room temperature tosolidify the synthetic rubber.7. Post-treatment: The solidified synthetic rubber may require further processing steps like washing, drying,and cutting into desired shapes or sizes.8. Quality control: Test the synthetic rubber for its physical and chemical properties to ensure it meets therequired specifications. This may include testing for tensile strength, elongation, hardness, and other relevantparameters.Note: The exact process conditions and additives used may vary depending on the specific requirements andintended applications of the synthetic rubber being produced. It is always recommended to follow establishedindustrial protocols and safety guidelines when working with chemicals and conducting polymerizationreactions.NOTE: LEWIS DIAGRAMS DO NOT TRANSCRIBE CORRECTLY. BUT THE IDEA IS HELPFUL.What is the Lewis diagram for styreneTo draw the Lewis diagram for styrene (C8H8), we first need to determine the number of valence electrons foreach atom. Carbon (C) has 4 valence electrons, and hydrogen (H) has 1 valence electron.Styrene consists of a benzene ring (C6H6) with an attached ethenyl group (C2H3). The benzene ring hasalternating double and single bonds, while the ethenyl group has a double bond.Here's the Lewis diagram for styrene:H|H ! C = C ! H|C/ \C C/ \H HIn this diagram, the double bonds are represented by two lines, while the single bonds are represented by aCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.12 Research Paper - Regroup111 single line. The carbon atoms in the benzene ring are connected by single bonds, while the carbon atoms inthe ethenyl group are connected by a double bond. The hydrogen atoms are attached to the carbon atoms.Note: The Lewis diagram is a simplified representation of the molecule's structure, showing only the valenceelectrons and the connections between atoms. It does not show the three-dimensional shape of the molecule.What is the Lewis diagram for butadieneThe Lewis diagram for butadiene (C4H6) can be represented as follows:H H H H\ / \ /C=C=C=C| |H HWhat is the difference between a dieneand a butadieneA diene is a hydrocarbon compound that contains two double bonds in its structure. It is a general term used todescribe any molecule with two double bonds, regardless of the number of carbon atoms present. On the otherhand, butadiene is a specific type of diene that contains four carbon atoms and two double bonds. In otherwords, all butadienes are dienes, but not all dienes are butadienes.Describe the chemical reaction between butadiene and styrene to form synthetic rubberThe chemical reaction between butadiene and styrene to form synthetic rubber is known as copolymerization.It involves the combination of monomer molecules of butadiene and styrene to create a polymer chain.The reaction is initiated by a catalyst, typically an organometallic compound such as a lithium or nickelcatalyst. The catalyst helps in breaking the double bonds present in the monomer molecules and facilitates theformation of new bonds.During the reaction, the double bond in butadiene undergoes a process called addition polymerization. Thedouble bond is opened up, and the carbon atoms on either side of the double bond attach to the monomerunits of styrene. This results in the formation of a long polymer chain with repeating units of butadiene andstyrene.The copolymerization of butadiene and styrene allows for the incorporation of different properties from bothmonomers. Butadiene contributes flexibility and resilience to the synthetic rubber, while styrene adds strengthand rigidity. This combination of properties makes the resulting synthetic rubber suitable for variousapplications, such as tire manufacturing, automotive parts, and industrial products.Overall, the chemical reaction between butadiene and styrene to form synthetic rubber involves the addition ofmonomer units through copolymerization, resulting in the creation of a polymer chain with desirable propertiesfor rubber production.Does styrene have a vinyl group in itYes, styrene does have a vinyl group in its chemical structure. The vinyl group is a functional group consistingof a carbon-carbon double bond (C=C). In the case of styrene, the vinyl group is attached to a benzene ring.Does the chemical compound butadiene Contain a vinyl groupYes, the chemical compound butadiene contains a vinyl group. The vinyl group is a functional group consistingof a carbon-carbon double bond and a single bond to a hydrogen atom. In butadiene, there are two vinylCI Laboratory Notebook Vol XXXIII : Appendix (Digital Format)2023.12.12 Research Paper - Regroup112