Search

Cellulase and pentosanase were largely separated from polygalacturonase in the crude preparation, Enzyme 19-D by paper chromatography. A partial separation of pentosanase from other enzymes of the mixture was effected by paper electrophoresis. All of the enzymes studied except cellulase were inactivated by heating at 74–75 °C.; almost 50% of the cellulase activity was retained. Polygalacturonase and pectinesterase were inactive above pH 7.0. Most of the enzymes studied appeared to have two or more components.

Paper chromatography of phenols on polyamide impregnated paper was devised. Fourteen phenols in seven different solvents were studied. The Rf values were tabulated. The relation between the structure of phenols and Rf values was discussed. This method was applied for the identification of phenolic constituents of Chamaecyparis formosensis Matum, in wich p-cresol, o-cresol and carvacrol were identified.

Modified paper chromatography procedures for the analysis of fatty acids produced by anaerobic bacteria are described. Both ethylamine and hydroxylamine derivatives of fatty acids were prepared from inoculated anaerobic culture broth. The derivatives were spotted on chromatography paper and developed with appropriate solvents. Paper chromatography is a valuable alternative to gas liquid chromatography as an ancillary procedure in the identification of anaerobic bacteria in the clinical bacteriology laboratory.

1. (1) The binding of acid dyes by denatured protein on filter paper has been investigated with special reference to Light Green. 2. (2) The uptake of dye, measured by elution, has been shown to vary according to the method of denaturation, the nature of the dye solvent (particularly the salt and and ethanol concentration) and finally with the duration and temperature of the subsequent washing procedure. 3. (3) Albumin and globulin do not behave in the same manner with different denaturing agents so that different methods give different albumin/globulin dye uptake ratios. 4. (4) The stoichiometric relationship between dye and protein is consistently nonlinear even at lower concentrations of protein on the paper and the same pattern can be demonstrated with protein on cellulose acetate membrane. It is suggested that this non-linear relationship at low protein densities is due to interference by the cellulose supporting medium and other anions. 5. (5) Intrinsic constants and the number of binding sites at infinite dye concentration have been measured under standardized conditions for Light Green, Brom-Cresol Green and Lissamine Green. The results have been compared and differences discussed. 6. (6) In the quantitative measurement of protein based on dye uptake, the importance of restricting the protein density on the paper is stressed.

1. 1. An established method of chromatography and polychromatic staining of phospholipids on untreated paper has been more fully investigated with respect to sulphatides. 2. 2. A rapid staining method for sulpholipids on paper chromatograms is described. 3. 3. The optical properties of spherical aggregates of the sulphatides under crossed polarizers are noted. 4. 4. Sulphatides have been shown, by chromatography, to occur in many tissues including those of rat, mouse, ox, reptile, certain marine animals, Aplysia, Archidoris, Coryphella, in some microsomal fractions and in lipovitellin. Sulpholipids also occur in fungi.

In order to effectively control slime production in paper plants and other major water consuming industries, the efficacy of selected chemical compounds as inhibitors of growth of Sphaerotilus natans was investigated. It was determined that a few products were effective against Sp. natans in paper mill process water (80°F). These are listed in the following order of decreasing effectiveness: chlorine, Busan 90, chlorine dioxide and Slimacide V-10. In vitro studies can suggest those chemicals which may be useful; however, the choice of inhibitors depends largely on the individual plant, i.e., the mill processes' capacity to tolerate the concentration, contact time or cost of the specific chemical.

Paper chromatography is very helpful in the identification and comparison of antibiotics, but very few results from the application of this technique have been published so far. The awakening of interest in the search for antibiotics which are active against fungi pathogenic to plants and animals makes the investigator aware of the scant information that is available to help characterize such agents. A knowledge of the Rf values of the known antifungal agents would help advance the search for such therapeutic materials. Often these values, in conjunction with the biological inhibitory activities, are the only criteria by which one can determine whether the unknown agent appears to be a new material. When the Rf data are obtained for shake cultures, one can eliminate many previously described antibiotic-producing organisms from further study, thus allowing more research to be applied to antibiotics which are new and different. The studies reported in this paper should serve as a guide to the identification of some of the known antifungal antibiotics.

1 A new diagonal electrophoretic technique for determining the order of the lysine-containing tryptic peptides of a protein is described. The protein is converted into its trifluoracetyl derivative, digested enzymatically (or chemically), and the resulting peptides separated by paper electrophoresis. The paper is then treated with ammonia vapour, which re-exposes the ɛ-amino groups of the lysine residues, and submitted to a second electrophoresis at right angles to the first direction. Peptides containing lysine residues, together with the N-terminal peptide of the protein, are found to lie off a diagonal formed by all other peptides, whence they may be readily purified. A study of these peptides enables the order of the lysine-containing tryptic peptides in the protein to be deduced. 2 The technique has been successfully tested with insulin. 3 When the method was applied to porcine pepsin, the four tryptic peptides isolated were easily ordered and the carboxyl-terminal sequence of the protein shown to be Arg-Gln-Tyr-Tyr-Thr-Val-Phe-Asp-Arg-Ala-Asn-Asn-Lys-Val-Gly-Leu-Ala-Pro-Val-Ala. The three basic amino acid residues in the molecule are thus found clustering towards the C-terminus of the polypeptide chain. 4 A common ancestral gene for porcine pepsin and bovine (calf) rennin is suggested by the close homology between the C-terminal sequences of the two proteins.

A critical review of the paper chromatography of Strychnos alkaloids has led to the following conclusions: 1. Paper chromatography is one of the most important methods of approach in elucidating the chemistry of Strychnos alkaloids. 2. Paper chromatography and electrophoresis, in the case of Strychnos tertiary alkaloids, are of great value for the rapid analytical identification of the various compounds. In the case of quaternary alkaloids these methods can give general information, but no absolute conclusions, about a great number of alkaloids so far known of this group, and facilitate the identification of some of the more common bases. 3. No absolute relation between RM value and structure can so far be derived from the RC values of Strychnos alkaloids. This is due to the extremely complicated structures of these alkaloids, which belong to several chemical groups, and to the difficulty of obtaining standard RC values for many quaternary alkaloids. 4. Paper electrophoresis of Strychnos tertiary alkaloids is of great importance for the identification of the different strychnine derivatives. 5. Paper chromatography and electrophoresis must be considered as the best methods hitherto used for the preparative purification of small quantities of Strychnos quaternary alkaloids.

Thermotolerant fungi have been found in slimes at temperatures of 38 to 52 °C in paper mills in Eastern Canada. The growth of some of these isolates on malt agar plates maintained at temperatures of 10 to 65 °C has been examined. Optimum temperatures varied between 30 and 50 °C, depending upon the species. Thiamine deficiency was noted in Trichosporon capitatum and Mycelium sterilium. The requirement of T. capitatum for this vitamin could be met by incorporating the thiazole and pyrimidine moieties together in the medium.

Mayor Gamble has just spoken to you on water-supply from the standpoint of the health of citizens. May I discuss with you water from the standpoint of a chemist as it is required by industrial concerns as the most necessary feature of manufacturing operations, especially in the manufacture of pulp and paper, which promises to be the next great industrial development in this southeastern territory which is governed by your membership. A recent survey by the Federal Government has shown an enormous wealth of pine and semi-hardwood timber in this practically untouched region. The figures of this survey are now attracting nation-wide attention, especially since the spruce woods of the Great Lakes and Eastern States are practically exhausted and this makes necessary the importation annually of some two hundred million dollars worth of pulp and paper. About fifteen years ago when it was first proposed to manufacture kraft from southern pine it was loudly proclaimed that this was impossible, but during the intervening years there has been so rapid a growth of the kraft pulp and paper industries that the South now dominates the field with the manufacture of more than sixty percent of the national output. Now when it is proposed to use southern pine for white paper the same cry is heard about the unsuitability of southern pine. The work in the Pulp and Paper Laboratory of the Industrial Committee of Savannah has completely disproved this contention. Erroneous ideas have been held regarding our pines: first, through the use of the name "yellow" pine, which applies only to the heart, while the sapwood gives a pulp actually lighter in color than spruce; and, second, the utilization of longleaf and slash pines for naval stores

Paper used for Japanese umbrella is vat-paper as well as machine-paper and oiled these Japanese paper, which is chiefly made from paper-mulberry.This report shows the superiority of those oiled-papers of various kind which are practically used after having examined their physical and chemical characteristics, and concluded as follows.(1) Japanese paper, if colored, increases its intensity.(2) Colored paper, when oiled, is not generally intensified, but, in case of white paper, it increases intensity.(3) To speak of oil, perilla oil or the one mixed up with a small quantity of paulownia oil is recommended.(4) When oil is twice coated, intensity is increased, but inferior oil such as substitute oil may not be very much beneficial even if twice coated.(5) As oiling method, hand-appling with waste-silk available at present is not abtained good results, it should be improved.

Chromatographic resolution of protein mixtures: egg albumin and lysozyme, and another containing insulin and protamine (Mugil cephalus) was achieved on paper strips using a several step elution with neutral salt solutions of increasing concentrations. Experimental data concerning the influence of cations and anions on chromatographic separation of proteins on paper strips are presented.

Differentiation of radioactive DNA and RNA deposited on filter paper discs can be accomplished by a relatively simple procedure. RNA can be efficiently removed by incubating the dises, impaled on pins, with 0.2 ml of 0.5 n NaOH for 90 min at 37°C. DNA can be removed after NaOH hydrolysis by treating the discs with 5% TCA for 30 min at 90°C. A correction is necessary to determine the actual amounts of DNA and RNA in order to account for the loss of DNA (13.8%) during the NaOH hydrolysis procedure.

A simplified procedure for filter paper electrophoresis is described in which disturbing factors such as evaporation, heating, buffer concentration gradients, and pH changes in the electrode vessels were reduced to a minimum. Artificial mixtures of highly purified proteins could be separated and the components isolated. The application of the method to a variety of studies on serum proteins is demonstrated. Protein concentration in paper segments was determined by two different methods of protein estimation. Curves were obtained showing the same five major peaks for normal serum as found by the classical methods of free electrophoresis. Comparisons were made of the areas of the various components under the curves obtained with the different methods. Two dimensional electrophoresis was applied to serum and serum components. It proved of value in demonstrating the heterogeneity of fractions such as the γ-globulin of serum. The polysaccharide dextran was used as an index of the extent of electro-osmotic flow during the course of the various experiments. The ratio of the distance of electroosmotic flow and the distance of protein migration was shown to be constant for a given type of paper. For serum albumin on Munktell 20 paper this ratio was 0.35. A formula for mobilities applicable to liquid in a highly porous supporting medium is presented. Mobility values for human serum albumin at various pH levels on paper showed approximate agreement with those obtained in free solution giving a similar isoelectric point.

Summary1. Paper electrophoresis provides a simple reproducible method for the study of serum (plasma) lipoproteins. Characteristic species lipoprotein patterns were demonstrated in normal man, monkey, dog and rabbit. 2. This method combined with chemical determinations of serum (plasma) lipids permits detailed investigations of normal and abnormal lipid metabolic states. 3. Elevation of serum (plasma) lipid fractions induced in these species by various experimental procedures were accompanied by a lowering of α lipoprotein and varying degrees of elevation of the β + O fraction.

Larvae of the silkworm, Bombyx mori L., were found to feed on filter paper which had previously been heated to approximately 200°C for 15 hr. Feeding on filter paper was largely accelerated when the paper had been impregnated with sucrose. β-Sitosterol and soybean sterol, previously demonstrated to stimulate feeding of the silkworm on agar-diets, showed rather little stimulating action on paper-feeding.

By the application of the crossing paper electrophoresis, antigen-antibody reactions could be detected on the filter paper. Antiserum was applied on a line on the anodic side of the filter paper and antigen was applied on a line drawn obliquely to the former on the cathodic side. By the electrophoresis, the antiserum was separated into its components and crossed over by the antigenic proteins. Antigen-antibody precipitates occurred in the zone of γ-globulin of the antiserum.The line of precipitate formed by the reaction of egg albumin with its homologous antiserum was dissolved and moved, if excess of egg albumin had crossed with it. The antiserum obtained in the earlier stage of immunization, however, showed a line of complex, which was not dissolved by the excess of egg albumin. The complex was suspected not to be precipitate. Thus it was inferred that the anti-egg albumin antibody of the earlier stage would be “incomplete”, which would become complete precipitin afterwards.The cross reactions of the anti-hen's egg albumin with heterogenous antigens, egg albumins of duck, goose, guinea hen, and quail, could also be deteced.By the crossing paper electrophoresis of bovine serum and its rabbit antiserum, at least 6 lines of pricipitate were detected. Two-dimensional application of the method was also carried out: The antiserum alone was first separated on a line in the first dimension. Then the antigenic bovine serum was applied on a line vertical to the first one. And the second electrophoresis was carried out to the 2nd direction vertical to the first one. The proteins of bovine serum migrated into the zone of γ-globulin of the antiserum to form lines of precipitate. By the two-dimensional technique, however, the sensibility of the method could not be raised. A comparison with the technique of Garbar's immunoelectrophoresis was made.The cross reactions of the anti-bovine serum rabbit antiserum with heterogenous antigens, human, swine and goat serum, were detected by the method.

SummaryNine serum protein fractionation experiments using the continuous flow paper electrophoresis technique were performed. Six experiments were performed using the serum collected from individual patients 24 and 48 hours after receiving therapeutic doses of I-131 and 3 using the serum of euthyroid individuals which was incubated in vitro for 1/2 hour at 37°C with I–131 labelled thyroxine prior to fractionation. Protein fractions obtained were identified by conventional paper electrophoresis and analyzed for radioactivity, total protein and cholesterol, the latter being taken as an index of lipid contents. Similar results were obtained for both in vivo and in vitro experiments. Beta globulins were found to be capable of forming thyroid hormone protein complexes to a larger extent than gamma globulins, but the largest amount of radioactive hormone was found as an alpha-2 globulin-thyroid hormone complex. A patient showing a marked abnormality in his plasma proteins showed the highest percent of radioacti...

In spite of many attempts to modify the filtrate reaction of Brdicka's cancer test, none of them gave better results than the original one. We already reported that in patients with acute leukemia the first maximum of Brdicka's filtrate wave was often higher than the second maximum. Such phenomenon presumably depends on the high contents of polysaccharides in molecules of these blood mucoproteins. In this paper further efforts were made to clarify the factors influencing on the shape of the protein double wave, and to find out the polarographic characteristics and clinical significance of electrophoretically separated mucoprotein with special reference to the wave form. Among many experimental conditions, the temperature in polarographic cells was found to be most sensitive factor to affect the protein wave shape, so that all experiments were carried out at the constant temperature (20°C±0.05). Serum mucoprotein was evidently separated into two fractions, M1 fraction an M2 fraction, by paper electroyhoresis in McIlvaine's buffer of pH 4, 4. These two fractions showed polarographically different shape each other. The M1 fraction, the most rapidly anodic moving one, showed left side elevation in shape; in other words, the first maximum was higher than the second maximum. On the contraly, M2 fraction showed right side elevation in shape; the second maximum was higher than the first maximum. Furthermore, it was very interesting from clinical point of view that the characteristic shape of M1 fraction was to some extent variable depending on diseases. From the clinical observation on 259 cases including 119 cases of cancer and 11 cases of leukemia, it was revealed that in neoplastic patients the wave shape of M1 fraction showed clearcut difference from those in normal; Δh (distance from the second maximum to the first maximum) of M1 fraction in neoplastic patients was larger than that in normal. Thus, the cancer detection rate has markedly elevated as compared with the original filtrate test, namely from 77% to 90%o in our study.

Filter paper electrophoretic analyses of serum were carried out in a series of eighteen proved cases of multiple myeloma, twenty-three normal controls and 130 patients with non-myelomatous diseases, and compared with the results of classical Tiselius electrophoresis and chemical fractionation. Abnormal electrophoretic patterns were found in all of the cases of multiple myeloma studied, and tall narrow spikes representing abnormal proteins with varying mobilities in the globulin range were demonstrated in 78 per cent. The latter were considered diagnostic of myelomatosis since similar patterns have not been described in any other disease and since none were observed in any of the present controls. In six of the eighteen cases the diagnosis was first established on the basis of these electrophoretic patterns and subsequently confirmed by bone marrow biopsy. In eight others they were merely confirmatory in previously diagnosed cases. Of special interest were the diagnostic patterns found in four patients in whom myelomatosis was not suspected. There were significant quantitative differences between the results obtained by filter paper and classical electrophoresis, and the results of both differed from those of chemical fractionation. However, the diagnostic peaks of myelomatosis were equally well demonstrated by both electrophoretic methods. It is concluded (1) that the electrophoretic patterns of serum obtained by both the filter paper and Tiselius methods exhibit highly characteristic peaks in a high proportion of individuals with multiple myeloma, (2) that such patterns are of great diagnostic value and (3) that the filter paper method offers the advantages of simplicity, low cost and speed, without any loss in sensitivity or specificity, which make it especially valuable for screening large numbers of specimens in patients suspected of having myelomatosis.

CHANG (1955) reported the lowering of the blood hemoglobin of chicks due to free gossypol feeding and postulated that free gossypol in the diet leads to interference with the synthesis of the proteins of the blood in these chicks. In view of the useful information that has been obtained in recent years using the electrophoresis technique in the study of normal and pathological sera, an experiment was planned to study the electrophoretic patterns of serum samples obtained from birds fed a normal diet, as well as from birds fed free gossypol in the diet. An electrophoresis cell using filter paper strips, as originally designed by Durrum (1950) and marketed by Spinco, was employed. Later it was discovered that the resolution of the globulins of chick serum is very poor on filter paper. More serum samples were obtained from birds used in a study of the effect of protein level on …

Summary 1. The literature on the use of paper chromatography in taxonomic work is reviewed briefly. 2. The technique of paper chromatography on disks is described and the method has been applied to a number of species of Lymnaea. 3. The chromatographic patterns of the species studied are described and discussed. 4. The implications of the discovery of species-specific substances in the body surface mucus of snails are discussed, both from the taxonomic and parasitological points of view. 5. The possibilities of applying two-dimensional chromatography to the free amino-acids of snail blood are discussed.

This study investigated effects of industrial wastes of a paper mill on fish. Chemical analyses were made of the effluent and of the river water before it entered the mill and at the effluent entry point. Static bioassay techniques were used to determine tolerances to the effluent of ten fish species which were abundant in the paper mill area. Fish were collected from the effluent entry point into the river. An abundant fish fauna was found. Charmin Paper Products Company, Cheboygan, Michigan, treats its waste products thoroughly and is an example of what an industry can do to reduce water pollution. The common shiner, Notropis cornutus , was found to be intolerant to even low concentrations of the effluent. Thus, its presence in an area would indicate a very low level of pollution of this type.

Paper chromatography of freeze-dried larvae of Mayetiola destructor (Say) in a solvent system of 1-butanol:methanol:water, 80:5:15, isolated a peak of plant-growth-promoting substance at Rt 0.15-0.25. This growth promoter was detected also on chromatograms of stomachs and carcasses of fresh larvae but was absent from chromatograms of larvae collected from ‘Knox 62’ wheat, Triticum aestivum L., a cultivar resistant to the Hessian fly, or from chromatograms of larvae that had never fed. Paper chromatograms of ethanol extracts of both infested and uninfested wheat plants revealed that feeding by the larvae did not reduce the amount of the growth promoter, but benzene extracts of infested plants produced chromatograms with more peaks of growth-inhibiting substances than did benzene extracts of uninfested plants. The inhibitors in the benzene extracts of infested plants may be the reaction of the wheat plant to secretions introduced into the plant by the feeding larvae. Therefore, the most likely cause of stunting of winter wheat by the Hessian fly is the result of something discharged into the plant by the Hessian fly larva as it feeds.

The technique of paper chromatography has been adapted to the identification of gluconic, 2-ketogluconic, and α-ketoglutaric acids. Combinations of methyl and ethyl alcohol were found to be the most suitable solvents and ammoniacal silver nitrate was found to give the most satisfactory reaction. When grown under normal physiological conditions where glucose was metabolized to carbon dioxide and water, Pseudomonas aeruginosa 9027 was shown to have oxidized glucose by way of gluconic and 2-ketogluconic acids. Since a strong system for oxidizing both gluconic and 2-ketogluconic acids was demonstrated, the presence of these acids over at least an eight hour period of growth is taken as evidence of their importance as intermediates in the oxidation of glucose by this organism.

Although chromatography is a comparatively old procedure, the method of paper partition chromatography did not become popular until well after 1944 when Consden, Gordon and Martin introduced it and demonstrated its application. This tool was given renewed interest and greater utility as a result of the development of the ascending method of paper chromatography by Williams and Kirby in 1948. Since that time, chromatographic procedures with paper have been applied by a host of investigators to natural, synthetic, organic and inorganic substances. Bacteriologists, geneticists, botanists, zoologists and a variety of others have employed paper chromatography as an analytical method for the study of proteins, amino acids, carbohydrates, steroids, antibiotics, vitamins and many other substances.

SUMMARY mitochondria were incubated in air in the presence and absence Methods are described for the resolution of coenzyme Q of substrates, the ultraviolet absorption spectra of the cyclo- homologues and other lipides by means of reversed phase paper hexane extracts of such particles were shown to change in a chromatography on silicone-impregnat,ed filter paper. manner which denoted a substrate-dependent reduction of coenzyme Q (9). Such extracts have now been chromatographed Acknowledgments-The authors are indebted to Dr. David on paper, and the results have been consistent with the previous E. Green for his encouragement in the course of these studies. interpretation of the spectra of these extracts. The data given Oscar Mayer and Company kindly supplied the large quantity in Fig. 2 indicate that the extract of particles incubated in the of tissue used in the investigation. REFERENCES

Thiolysis of an imidazolyl-dinitrophenyl-histidyl peptide at either pH 3.5 or 6.5 results in an increase in the net positive charge on the peptide. It is shown that this property can be used to form the basis of a diagonal paper electrophoretic purification of histidyl peptides from proteins. The amino groups of the protein are first reacted with citraconic anhydride and then the citraconyl protein is reacted with 1-fluoro-2,4-dinitrobenzene. The dinitrophenyl-citraconyl protein is digested with pepsin in 10% formic acid and, if necessary, with other proteolytic enzymes. The enzymatic digest is subjected to high-voltage paper electrophoresis at either pH 3.5 or 6.5. A guide strip is removed, thiolyzed with 2-mercaptoethanol, and subjected to electrophoresis at the same pH at right angles to the original direction of electrophoresis. The histidyl peptides are displaced off the diagonal toward the cathode. The off-diagonal peptides are isolated from the original electrophoretogram by thiolysis and electrophoresis using the diagonal electrophoretogram to locate the positions of the dinitrophenyl-histidyl peptides.

1) The daily change of the electrophoretic pattern of the silkworm blood from larval to pupal stage was examined. Three components (b3, b2, b1) were present in the blood of the early period of the 5th instar as shown in the previous paper (AIZAWA, 1955), while from 4-5th day of the 5th instar to pupal stage, there appeared two components, which were densely stained (Figs. 1, 3). In the latter case, when the electrophoresis was performed with the diluted blood, three protein components could be clearly detected (Fig. 2). b3 is the fastest moving component and it seems to be albumin. b2 and b1 are probably globulin. b3 component decreased with the pupal age and finally disappeared, at earlier time in male than in female.2) Comparison of the electrophoretic patterns was made by veronal, phosphate, citrate, borate or tris (hydroxymethyl) aminomethane buffer with the varying ion concentration and pH. The separation of the protein fractions with veronal buffer (pH 8.6, I=0.05) was better than with other buffers.3) Comparison of staining, was examined using bromphenol blue, solar blue black and amido black. All dyes were suitable for the purpose. By sudan black staining, b3 component was slightly stained (Fig. 4).4) Paper strips were cut off from the electrophoresis paper of the jaundice-diseased blood at the regions of b3, b2+b1 and the starting line as shown in Fig. 5. The virus was extracted with the distilled water and the injection experiments were performed. The virus amount was highest in b2+b1 fractions and it decreased in the fraction from near the starting line. The virus activity was scarecely shown in the fraction of b3.5) Any difference, except for quantitative one was not observed in the electrophoretic patterns of the blood between the normal and diseased larvae (both nuclear and cytoplasmic polyhedroses). On the contrary, the pattern of Galleria mellonella is, however, different from that of Galleria-adapted silkworm jaundice virus (Aizawa, unpub.).

In few conditions is the electrophorogram of serum proteins capable alone of deciphering the diagnosis. In conjunction with the assembled clinical data and relevant applied pathological tests the definition of the disease is often clarified. The patterns as observed by the applied pathologist are serially discussed and the importance of the variations in different fractions presented. In several instances, exemplified by liver disease, the assistance rendered toward prognosis has been considerable. Perhaps the most clearcut diagnosis is possible in the agammaglobulinaemia syndrome which is immediately evident by the technique of paper electrophoresis. Current advances are mainly in the direction of estimating the protein-bound carbohydrate and lipid constituents which are believed to reflect the abnormalities in several infections and metabolic diseases. Utilisation of non-ionic detergents has also allowed separation of subfractions of the globulins into numerous new components, including lipo-proteins, some of which had been previously observed during the application of improved buffer systems designed to restrict interference of heavy metals and to facilitate accentuation of fractions having intermediate mobility.

This paper presents the results of the application of circular filter paper chromatography to the differentiation of pathogenic and non-pathogenic staphylococci by means of an arginase test, and to the differentiation of haemolytic streptococci and viridans streptococci by an arginine dihydrolase test. Chromatographic assay indicates that in the group of staphylococci only the pathogenic and not the non-pathogenic staphylococci metabolize arginine to ornithine and urea (presence of arginase). Chromatographic assay indicates that in the group of streptococci only the haemolytic streptococci and not the viridans streptococci metabolize arginine to ornithine and citrulline, or to ornithine or citrulline only (presence of arginine dihydroase in the haemolytic group).

Total alkaloidal extracts from four closely related Lobelia species, L. cardinalis L., L. syphilitica L., L. puberula Michaux, and L. elongata Small, and ten crosses and hybrids involving L. cardinalis and L. syphilitica, were examined using two-phase descending paper chromatography with formamide-ammonium formate-formic acid as the stationary phase and chloroform-benzene as the mobile phase. The chromatogram of L. elongata showed the presence of lobinaline, suggesting its closer relationship with L. cardinalis. The chromatograms of L. cardinalis, L. syphilitica, and L. puberula were different from each other, suggesting the elaboration of different alkaloids by each of these related species. Lobinaline was present in the first species and absent in the latter two species. The L. cardinalis × L. syphilitica hybrids gave chromatograms suggesting the elaboration of alkaloids characteristic of both the parent plants.

An indirect immunofluorescence test using small discs (0.4 cm in diameter) cut from blood smears on filter paper is described. Discs plus three drops of phosphate-buffered saline are laid on antigenic areas on slides, which are then incubated. During incubation, eluted antibodies react with antigen. The method was tested with 10 eluates from healthy persons and with 30 eluates from three groups of 10 patients with Chagas’ disease, schistosomiasis Mansoni and South American blastomycosis, respectively. Sera from the same blood samples were tested also by conventional indirect immunofluorescence tests. Both tests gave similar results. The method proposed is useful as a rapid qualitative screening test when dealing with a large number of samples.

(1952). Reproducibility with Paper Electrophoresis of Serum Proteins. Scandinavian Journal of Clinical and Laboratory Investigation: Vol. 4, No. 4, pp. 307-312.

An application of the biuret reaction to the determination of protein fractions on filter paper electropherograms of serum is described. The relative mobilities of the serum protein fractions of the domestic fowl and of man are compared. Values are reported for serum protein fractions as separated by filter paper electrophoresis in a methanolic veronal buffer. Some observations on the serum proteins of the estrogenized immature pullet are reported; and it is tentatively suggested that another fraction as well as serum phosphoprotein appears in the serum of the pullet as a consequence of treatment with estrogen.

For making Japanese Style Paper, vegetable mucilage such as "Tororo-aoi", " Nori-asa" or "Ninori"is usuallyadded to obtain uniformpaper sheet. Mechanism of the uniformsheet formation property of mucilage is discussed principally on the standpoint of drainage of water.Drainage time has been measured by using upper part of schopper's freeness tester. Three kinds of vegetable mucilage extracted from "Tororo-aoi" (Abelmoschus manihot medic), "Nori-asa" (artificial couple of "Tororo-aoi" and "Okura " Abelmoschus esculentus Moench made by prof.Kagawa) and "Ninori" (Hydragea paniculata sieb) have been studied.Effects of the amounts and the kinds of vegetable mucilage, pulp consistency and temperature of water.Drainage time increases according to the increase of the addition of vegetable mucilage or pulp consistency, and decreases by rising the temperature of water.That is to say the drainage degree is the function of (1) the added amount of vegetable mucilage (2) pulp consistency (3) resistance of fiber layer (4) resistance of filtering material (5) beating degree of pulp, and the reverse function of (6) the temperature of water (7) filtering area (8) head difference in vat and (9) x (etc.) : Drainage =Kφ {(the added amount of vegetable mucilage) (pulp consistency) (resistance of fiber layer) (resistance of filtering material) (beating degree of pulp) (the temp. of water) (filtering area) (head difference in vat) (x)}.By using white water repeatedly, effect of vegetable mucilage and also drainage time decrease gradually. In the case of using white water repeatedely, the recovery percentage of available component of vegetable mucilage measured by the drainage time, is about by 20% "Tororo-aoi", by 40% "Nori-asa", and by 60% "Ninori ". The order of depression of viscocity of vegetable mucilage is as follows, "Tororo-aoi", " Nori-asa " and " Ninori ". Therefore in mill practice the percentage of recovery of available component of " Ninori " has become greater than the above mentioned data. This is the reason why the " Ninori " notwithstanding the mucilagious extract is smaller than "Tororo-aoi", is comparatively favourable in mill practice.

This paper describes a paper chromatographic method of identification of morning glory (Ipomoea purpurea) seeds used by individuals seeking hallucinatory experiences. The procedure is applicable to a single seed, seed fragments and products of “tea”. A 50 percent aqueous alcoholic extract is chromatographed in a butanol : acetic acid : water system and the pattern visualized by ultraviolet fluorescence, p-dimethylaminobenzaldehyde and ninhydrin spray reagents. The chromatographic patterns of various morning glory seeds and other seeds of criminalistic interest are shown. Milligram quantities of Ipomoea purpurea (two varieties) are readily differentiated from all other seeds examined.

SUMMARY: A simple method of detecting arabinose by paper chromatography in the cell-walls of aerobic actinomycetes is described. Mycobacteria and Nocardia species are rich in arabinose while saprophytic species of Streptomyces are deficient. Pathogenic Streptomyces species tend to fall between the two extremes but Streptomyces somaliensis is apparently nearly devoid of arabinose.

High rates of acetylene (C 2 H 2 ) reduction (nitrogenase activity) were observed in woodroom effluent from a neutral sulfite semi-chemical mill under aerobic (up to 644 nmol of C 2 H 4 produced per ml per h) and under anaerobic (up to 135 nmol of C 2 H 4 produced per ml per h) conditions. Pasteurized effluent developed C 2 H 2 reduction activity when incubated under anaerobic but not under aerobic conditions. Activities were increased by addition of 0.5 to 3.0% glucose or xylose. Enrichment and enumeration studies showed that N 2 -fixing Azotobacter and Klebsiella were abundant, and N 2 -fixing Bacillus was present. Of 129 isolates of Klebsiella from pulp mills, lakes, rivers, and drainage and sewage systems, 32% possessed nitrogen-fixing ability.

A convenient, sensitive, and reliable assay for the conversion of radiolabeled sucrose to alcohol-insoluble dextran by the Streptococcus mutans dextransucrase has been developed.

A radiochemical method for the direct measurement of thiamine pyrophosphokinase (ATP: thiamine pyrophosphotransferase, EC 2.7.6.2) activity was described earlier (1,2). It avoided the difficulties associated with assay systems based on the coenzyme nature of thiamine pyrophosphate in TPP-dependent 1 enzyme reactions using apopyruvate decarboxylase (3) (2-oxoacid carboxylase, EC 4.1.1.1) or apotransketolase (4) (sedoheptulose-7-phosphate: d -glyceraldehyde-3-phosphate glycolaldehydetransferase, EC 2.2.1.1). Since the chromatographic isolation of TPP is time-consuming, a procedure for the rapid determination of thiamine pyrophosphokinase activity was desirable. The simplified method described here takes advantage of the anionic character of TPP. The assay is carried out with [ 14 C]thiamine as substrate. After incubation with the enzyme in the presence of Mg 2+ -ATP, the reaction mixture is applied to a DEAE-cellulose paper disc. The disc is extensively washed with sodium acetate resulting in the quantitative elution of [ 14 C]thiamine and partial retention of [ 14 C]TPP. This is quantitatively measured using the liquid scintillation counting technique. A similar procedure has been described for the determination of glycerol kinase (ATP: glycerol phosphotransferase, EC 2.7.1.30) and hexokinase (ATP: d -hexose 6-phosphotransferase, EC 2.7.1.1) activities (5).

1. A modification of the usual paper electrophoretic procedure has been made by including in the buffer medium 0.05 per cent of certain alkylphenyl ethers of polyethylene glycol. 2. This modified procedure yields electrophoretic patterns of blood serum proteins that may be subdivided into 12 fractions— albumin, α1-globulin, three α2-globulins, three β-globulins, and four γ-globulins. 3. The method was applied to blood serum from 70 patients apparently free of significant disease and to sera from a number of patients with varied disease. More specific location of the altered protein levels was possible in a number of disease states. 4. Glycoprotein distribution was not seriously disturbed by the presence of the nonionic surfactants, but lipoproteins migrated at a decreased rate.

An investigation of the electrophoretic data from patients with various malignant diseases shows that the possibilities of paper electrophoresis as a diagnostic tool are limited. The relative inaccuracy of the method may cover up important details; but more physiological information can scarcely be expected from data concerning only one physical property of a protein molecule. Quantitative evaluation of the data still gives rather qualitative results. With more material it may be possible to differentiate between various groups of diseases. The use of the “entropy” of a serum protein mixture does not give greater differentiation. Only in a limited number of cases can it be used to distinguish pathological and normal sera. The high correlation in the normal group between the albumin contribution to the total “entropy” and the total “entropy” provides a criterion for the detection of pathological serum protein mixtures in most cases.

In order to determine the effective shoot cutting height on the high training in paper mulberry, two experiments were conducted. The results were as follows. Cutting at the height higher than 15 to 30 centimeters from ground surface in the first year, showed the merits as increased yield and yield rates and the distinguished marks of high training. The most effective shoot cutting height on and after the second year was 3 centimeters in 30 centimeter high training.

1. 1. The elctrophoretic mobility in filter paper at pH 8.6 was measured for hemoglobins from seventy-six rodents representing fifteen species or subspecies from the following genera: Phyllotis, Akodon, Chinchillula, Mus, Hesperomys, Oryzomys, Ctenomys and Cavia . 2. 2. These rodents showed a wide range of hemoglobin mobilities from 0·6 to 1·5 times that of man (HbA). 3. 3. Within the genera Phyllotis and Akodon the range in subgroups was 0·63–1·15 and 0·62–1·24 respectively. 4. 4. Secondary components, usually slower, were common. 5. 5. No general correlation of mobility with oxygen affinity (high-altitude habitat) was found, but correlation in several case within species or genera. 6. 6.Alkali-denaturation resistance ranged from 5 to 86 per cent and showed some taxonomic differences between subgenera. 7. 7. Some observations on two Camelidae (alpaca and vicuna) confirmed the low mobilities seen this group.

Single and two-dimensional paper chromatograms were prepared of normal and de-coated embryonated eggs of Ascaris suum, vitelline membranes, egg fluid, and larvae. Eighteen amino acids (cystine, lysine, arginine, histidine, aspartic acid, glycine, serine, alanine, proline, glutamic acid, threonine, tyrosine, methionine, valine, tryptophan, phenylalanine, leucine, isoleucine) were identified in larvae and in normal and decoated embryonated eggs. Nine amino acids (leucine, tryptophan, cystine, isoleucine, phenylalanine, tyrosine, proline, aspartic acid, and arginine) were identified from the vitelline membrane. Seven amino acids (cystine, arginine, aspartic acid, glycine, serine, proline, tyrosine) were identified from a fraction in the middle coat. Using the DNFB technic, the N-terminal amino acid in the vitelline coat was determined to be cystine. Sixteen amino acids (cystine, lysine, histidine, aspartic acid, glycine, serine, glutamic acid, threonine, proline, tyrosine, methionine, valine, leucine, tryptophan, phenylalanine, isoleucine) were identified from the outer protein coat.

I investigated amino acids composition of the protein from the tuberculous pus plasma in 3 cases and free amino acids in the tungstic acid filtrates of tuberculous pus plasma by a paper-chromatographic procedure. 1) I found tuberculous pus plasma proteins do not essentially differ from usual proteins regarding the qualitative composition. 2) In 3 cases of closed tuberculous abscess never punctured free amino acids were not detected. 3) Free amino acids were present in the protein-free filtrates of tuber-culous pura of 5 closed abscesses often punctured before. 4) The number of the free amino acids found there, however, far less than that of the corresponding amino acids in hot true pura. There is a striking contrast between tubercuous and non-tuberculous true pura.

Auxins and auxin inhibitors from tissue extracts of normal Nicotiana plants, Nicotiana glavca, N. langsdorffii and their hybrid (which spontaneously produces tumors) were separated by ascending paper chromatography with n-butanol-distilled water. An Avena curvature test was used for demonstrating growth-promoting and growth-inhibiting substances. IAA could be found in extracts of the parents and the hybrid (RF 0.75). Hybrid tissue yielded the highest amount (37.10), N. glauca tissue less (30.80), and N. langsdorffii tissue the least amount (8.50) of IAA. A second growth promoter (RF 0.35) could be separated from the tissue extracts of the parents and the hybrid, but it showed only low activity in the Avena test. Three inhibitors were present in extracts from N. langsdorffii and the hybrid at RF 0.25, 0.45, and 0.85, whereas N. glauca showed only two of them (RF 0.25 and 0.85). The inhibitor with an RF of 0.45 seemed to be identical with the acidic, benzene-insoluble "inhibitor i" of Bennet-Clark and Kefford (1953). The inhibitor (neutral, benzene-soluble) at RF 0.85 could be found in some tissue extracts of the parents and the hybrid, but showed only little activity in the curvature tests. From neutral and from acidic plant extracts within a pH range of 4.4 to 5.8 a third inhibitor with an RF of 0.25 could be separated. It seems that the high concentration of natural IAA in the hybrid is regulated by a variety of inhibitors with different specificities in the growth-regulating process. Nicotiana langsdorffii tissue has much less auxin but the same variety of inhibitors as the hybrid, whereas N. glauca tissue contains less auxin than the hybrid and only two of the three inhibitors found in N. langsdorffli and hybrid