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Tonicity-volume relations in partially hemolyzed hypotonic systems.

The linear relation between the red cell volume V and the reciprocal of the tonicity T of a hypotonic medium is not the linear one expected on the basis of the van't Hoff-Mariotte law, particularly when a fraction p of the cells are hemolyzed and the volume V/(1 - p) of the (1 - p) cells which remain intact is considered. In systems of relatively high tonicity in which p is zero, the relation is linear but its slope is usually too small; at lower tonicities in which p has a value between zero and 0.35, the volumes are larger than those expected on the basis of the van't Hoff-Mariotte law, while at still smaller tonicities they are much smaller than expected. The volume measurements referred to are made with a high speed hematocrit; the results obtained in systems containing relatively high and relatively low volume concentrations of cells are contrasted with each other, and allowance is made in the calculations for the volume of the hypotonic medium surrounding the cells being limited. Attempts are made at an explanation of the anomalous results in terms of incomplete packing, of a stepwise as opposed to an all-or-none loss of Hb from the cells, of a heterogeneity in the swelling properties of the cells of the population, of a loss of osmotically active substances from the intact cells, and of the red cell ghost having some degree of rigidity. These explanations are not satisfying, although some of them in combination may account for the phenomena observed. It seems likely that the structures involved in the hemolytic process (cells and ghosts) have different properties in different tonicity ranges and even when the same tonicity is established in systems which are dense, as opposed to dilute, with respect to cell concentration. The form of the tonicity-volume relation can be changed substantially, although not in the direction of greater linearity, by treating the cells with resorcinol, colloidal silicic acid, iodoacetate, or sodium oxalate. Small changes in pH, exposure to temperatures as high as 37 degrees C., allowing the cells to stand for periods up to 24 hours, or the substitution of hypotonic plasma for hypotonic NaCl-buffer, produce, on the other hand, only minor changes in the tonicity-volume relation.

Cell Death↗

Hematoxylin substitutes. A study of phenocyanin TC and the use of afterchrome mordanting in histology.

A practical, simple synthesis of the obsolete mordant dye, phenocyanin, was devised, proceding from gallocyanin and resorcinol with acid and heat. The dye gave promise of good performance in metachrome iron mixtures, but because of excessive precipitation, the practice of afterchroming was taken from textile dyeing usage, and proved very successful. Of a number of metallic salts tried, Fe II proved to be the best, then Cu II and Fe III. The stain acts as a cationic dye on nucleic acids and other acidic tissue components: acid mucins, cartilage, mast cells, corpora amylacea, etc. The afterchroming process renders the stain much more resistant to various extraction agents and even moderately resistant to acid alcohol. Color values are quite comparable to those obtained with hematoxylin-eosin when an eosin counterstain is used. Nuclei basophilic cytoplasm, Nissl granules, and bacteria color dark blue; cartilage, mast cells, and some acid mucins, deep violet. Staining with the 1% solution was essentially unaltered from neutrality down to 1.2 N HCl, pH 0.68, and dilution of the dye to 0.05% in 1% conc. HCl (pH 1.2) still gave excellent nuclear, RNA, and mast-cell staining. At 0.02% and pH 1.2, nuclear staining was distinctly weakened; mast cell granules were still dark violet.

Benzopyrans↗

Occupational dermal exposure to permanent hair dyes among hairdressers.

Skin exposure to permanent hair dye compounds was assessed in 33 hairdressers using a previously evaluated hand rinse method. Hand rinse samples were collected from each hand before the start of hair dyeing, after application of the dye and after cutting the newly-dyed hair. Sixteen of the hairdressers did not use gloves during dye application, and none used gloves while cutting the dyed hair. The samples were analysed for pertinent aromatic amines and resorcinol (RES) using an HPLC method. 10 of 54 hair dye mixtures contained 1,4-phenylenediamine (PPD), 40 toluene-2,5-diaminesulphate (TDS), and 44 RES. After application of the hair dye, PPD was found in samples from 4 hairdressers, TDS in 12 and RES in 21. PPD was found in samples from 3 of the 17 hairdressers that used gloves during application of the hair dye, TDS in 5 and RES in 11. In the group that did not use gloves during the application of hair dye (n = 16) PPD was found in samples from 1 hairdresser, TDS in 7 and RES in 11. After cutting the dyed hair, PPD was found in samples from 5 hairdressers, TDS in 14 and RES in 20. Analysis of samples of newly-dyed hair cuttings revealed the presence of aromatic amines and/or RES in 11/12 samples. Our conclusion is that hairdressers' skin is exposed to allergenic compounds during hair dyeing. Exposure occurs from dye application, from cutting newly-dyed hair and from background exposure. The exposure loadings are in the level, where there is a risk of sensitization and/or elicitation of contact allergy (i.e. for PPD 22-939 nmol per hand). The glove use observed in this study was often improper, and was insufficient to prevent exposure. To reduce exposure, improved skin protection and work routines are important.

Adult↗

Comparison of the effects of 13 phenolic compounds in induction of proliferative lesions of the forestomach and increase in the labelling indices of the glandular stomach and urinary bladder epithelium of Syrian golden hamsters.

Groups of 6-week-old male Syrian golden hamsters were given 13 different phenolic compounds for 20 weeks. Of these compounds, 2(3)-tert-butyl-4-methoxyphenol (BHA), 2-tert-butyl-4-methylphenol (TBMP) and p-tert-butylphenol (PTBP) strongly induced hyperplasia and tumorous lesions in the forestomach. Catechol, p-methylphenol (PMYP), p-methoxyphenol (PMOP), caffeic acid, methylhydroquinone (MHQ) and pyrogallol were less active, and resorcinol, hydroquinone, propylparabene and tert-butylhydroquinone (TBHQ) were not active. The labelling index in the forestomach epithelium was significantly increased by addition to the diet of BHA, TBMP, catechol, PMOP, PTBP and MHQ. PMOP induced epithelial damage and regenerative hyperplasia of the pyloric region. Catechol, caffeic acid and PMYP induced similar though less marked lesions. The labelling index in the glandular stomach was significantly increased by oral catechol (P less than 0.05) or PMOP (P less than 0.05). No histopathological lesions were observed in the urinary bladder epithelium, but propylparabene (P less than 0.05), catechol, TBHQ and MHQ increased the labeling index. These findings indicate that PTBP and TBMP may be carcinogenic for hamster forestomach after long-term administration, and that both one hydroxy and tert-butyl substituents may be important for induction of hamster forestomach tumors.

Animals↗

A high-performance liquid chromatographic determination of major phenolic compounds in tobacco smoke.

A high-performance liquid chromatographic (HPLC) method is developed that simultaneously quantifies the dihydroxy compounds hydroquinone, resorcinol, and catechol and the monohydroxy compounds phenol, m + p-cresol and o-cresol in cigarette smoke. Particulate matter samples collected on Cambridge pads and in impingers by conventional trapping techniques are simply (no derivatization required) subjected to reversed-phase gradient liquid chromatography. Samples of both mainstream and sidestream smoke can be analyzed. Selective fluorescence detection is used to monitor the mobile phase effluent, by which these phenolic compounds are detected in the nanogram range. The detector response is linear, overall precision is good, and recoveries are greater than 95 percent. The total run time, excluding extraction, is one hour. The procedure has been applied to tobacco products whose smoke contains varying amounts of these phenols. Kentucky Reference Cigarette 1R4F was found to contain substantially more of these compounds than a new cigarette that heats but does not burn tobacco (New Cigarette). The method is compared with other procedures used to determine phenolics in cigarette smoke.

Chromatography, High Pressure Liquid↗

Massive in vitro synthesis of tagged oligosaccharides in 1-benzyl-2-acetamido-2-deoxy-alpha-D-galactopyranoside treated HT-29 cells.

Permanent exposure of differentiated HT-29 cells to the sugar analogue, 1-benzyl-2-acetamido-2-deoxy-alpha-D-galactopyranoside (GalNAcalpha- O -bn) leads to marked effects upon the phenotypic properties of mucin-secreting or enterocyte-like HT-29 cells: an inhibition in the secretion of mucins, a blockade in the apical targeting of membrane brush border glycoproteins and a swelling of cells with intracellular accumulation of numerous vesicles. Folch extraction and partition of treated enterocyte-like HT-29 cells revealed a very important accumulation of orcinol and/or resorcinol reactive material in the upper phase (usually containing gangliosides), as compared with untreated HT-29 cells and with treated and untreated Caco-2 cells. Structural analysis indicated the accumulation of a series of GalNAcalpha- O -bn derived oligosaccharides, most of them with the common core Galbeta1-3GalNAcalpha- O -bn. These oligosaccharides contained residues of GlcNAc, Gal, Neu5Ac, or Fuc. In particular, the tagged sialyl-Lewis(x)was identified, as well as more complex sialylated derivatives, including the sialyl-Lewis(x)substituted by an additional Neu5Ac residue. The benzylated oligosaccharides were not significantly detected in the culture medium except for Galbeta1-3GalNAcalpha- O -bn. Upon reversion of the treatment, these derivatives dis-appeared from the cells within few days, however were not recovered as such in the culture medium. Intracellular degradation occurred with desialylation and defucosylation as the first steps. The spectacular accumulation of benzylated oligosaccharides in HT-29 cell, permanently exposed to GalNAcalpha- O -bn very likely plays an important role in the alterations of cellular processes previously described in this cell line. The HT-29 cell culture system also appears to be an efficient source of several tagged oligosaccharides.

Cell Differentiation↗

Determination of beta2-agonists in bovine urine: comparison of two extraction/clean-up procedures for high-resolution gas chromatography-mass spectrometry analysis.

Two extraction/clean-up analytical procedures were investigated and compared regarding their recovery and matrix-purification efficiency for screening beta2-agonist residues in fortified bovine urine by high-resolution gas chromatography-mass spectrometry (GC-MS). The first procedure, based on an analytical method originally developed for detecting anabolic steroids, consists of the employment of the nonionic resin, Amberlite XAD-2, a styrene-divinylbenzene copolymer for solid-phase extraction (SPE), followed by liquid-liquid extraction with diethyl ether. The second focuses on the use of a mixed SPE cartridge (reversed-phase and ion-exchange sorbent, Bond Elut Certify). In both cases, the trimethylsilylated derivatives were analyzed by GC-MS with an ion-trap detector. Clenbuterol, salbutamol, and terbutaline were used to spike urine samples during the comparison experimental phase. Afterwards, tulobuterol, mabuterol, mapenterol, cimbuterol, and brombuterol were included in the evaluation of the second procedure (the Bond Elut Certify procedure). At this stage, the detection was accomplished by GC-MS (quadrupole mass analyzer) with selective ion monitoring acquisition. The isotopic dilution method with the hexadeuterated analogues of clenbuterol and salbutamol was applied to prepare calibration curves and calculate recovery percentages. With XAD-2 resin, terbutaline and salbutamol (resorcinol and phenol-type beta2-agonists, respectively) could not be detected at 20 ng/mL or at 40 ng/mL. In spite of clenbuterol having been detected at 20 ng/mL, the results obtained were not reproducible. The use of the reversed-phase and ion-exchange sorbent Bond Elut Certify allowed multiresidue detection and showed several advantages for the screening of clenbuterol such as higher recoveries, cleaner final extracts, reduced sample preparation time, less labor intensive, and easier solvent consumption and disposal. Recoveries over 88% (concentrations ranging from 0.5 to 10 ppb) and limits of detection equal to 0.5 ppb were met for all the beta2-agonists studied with the last method.

Adrenergic beta-Agonists↗

Urinary riboflavin metabolites in the rat.

Male rats were fed a riboflavin-deficient diet for 25 days and then fed [2-14C]riboflavin to replete their riboflavin pool(s). During a second depletion period, urine and feces were collected and 14C excretion monitored. The urine was chromatographed on R-15 resorcinol resin and 14C was measured in all fractions. The 14C compounds in the individual fractions were shown to be different by thin layer chromatography. The amount of 14C recovered as riboflavin increased after acid hydrolysis of the urine indicating the presence of riboflavin in conjugated forms. All R-15 fractions were found to support growth in a microbiological (L. casei) assay for riboflavin. This suggested that other derivatives, in addition to riboflavin and flavin nucleotides, are biologically active. However, the conjugated compounds were apparently less active, since acid hydrolysis of the urine enhanced its growth-promoting ability of L. casei. One 14C metabolite was identified as urea, thus providing evidence for degradation of the riboflavin molecule. This study has shown that the metabolic fates of riboflavin in the rat include conjugation and extensive degradation.

Age Factors↗

Cocarcinogenic and tumor-promoting agents in tobacco carcinogenesis.

A series of 21 tobacco smoke components and related compounds werere applied to mouse skin (50 female ICR/Ha Swiss mice/group) three times weekly with a low dose (5 mug/application) of benzo[a]pyrene (B[a]P). The test compounds were of five classes: aliphatic hydrocarbons, aromatic hydrocarbons, phenols, and long-chain acids and alcohols. The following compounds enhanced remarkably the carcinogenicity of B[a]P: catechol, pyrogallol, decane, undecane, pyrene, benzo[e]pyrene, and fluoranthene. The following compounds inhibited B[a]P carcinogenicity completely: esculin, quercetin, squalene, and oleic acid. Phenol, eugenol, resorcinol, hydroquinone, hexadecane, and limonene partially inhibited B[a]P carcinogenicity. Six of the 21 compounds were also tested as tumor promoters im two-stage carcinogenesis. No direct correlation existed between tumor-promoting activity and cocarcinogenic activity. The cocarcinogens pyrogallol and catechol did not show tumor-promoting activity. Decane, tetradecane, anthralin, and phorbol myristate acetate showed both types of activity. Structure-activity relationships and possible modes of action were described.

Acids↗

A prospective toxicity evaluation (COMPACT) on 40 chemicals currently being tested by the National Toxicology Program.

The computer-optimized molecular parametric analysis of chemical toxicity (COMPACT) procedure has been used to determine the molecular conformation and electronic structure of a series of 40 chemicals (out of a total of 44). The procedure can evaluate whether they interact with the active site of cytochrome P450 I or to the binding site of the Ah receptor, and hence to manifest carcinogenicity/toxicity. This is in response to the recent publication by Tennant et al. and their invitation to participate in a prospective identification of potential mutagenicity/carcinogenicity of these 44 chemicals. Correlation of COMPACT with potential genotoxicity was 25/40 (63%); COMPACT also predicted toxicity/carcinogenicity in 10 chemicals (25%) considered to be potentially non-genotoxic (naphthalene, promethazine, resorcinol, p-nitrophenol, tricresyl phosphate, bis(bromoethyl) propanediol, 3,4-dihydrocoumarin, theophylline, triamterene and chloramine), and predicted the absence of toxicity in four chemicals (10%) considered to be potentially genotoxic (methyl bromide, hydrazoic acid, 2,3-dibromo-1-propanol and 1,2,3-trichloropropane).

Binding Sites↗

The effect of polymerization of horseradish peroxidase on the peroxidase activity in the presence of excess H2O2: a background for a homogeneous enzyme immunoassay.

The phenol oxidation catalyzed by horseradish peroxidase (HRP) is slowed down by the presence of excess H2O2. This inhibition is due to accumulation of Compound III, which is a catalytically sluggish form of HRP. When HRP is polymerized through covalent bonds, Compound III becomes unstable and the peroxidase activity is less sensitive to excess H2O2. Under suitable experimental conditions, the phenol oxidation is increased by about 20-fold upon polymerization of the enzyme. This fact represents the principle of a homogeneous enzyme immunoassay reported by Hoshino et al. (J. Biochem. 97, 113-118 (1985)). The ratio of the peroxidase activities of monomeric and polymerized HRPs is 1 : 4 when phenol is replaced by resorcinol, and the difference is no larger when guaiacol and catechol are used as electron donors.

Horseradish Peroxidase↗

Water-soluble lipoproteins from yolk granules in sea urchin eggs. II. Chemical composition.

The chemical composition of yolk lipoproteins (YLP-1, 2, and 3) was determined. YLP-1, 2, and 3 were quite similar as regards the chemical composition of lipids, proteins, and carbohydrate moieties. Each lipoprotein has an average dry weight composition of lipids (55--72%) and apo-lipoproteins (28--45%) containing protein, hexose, hexosamine, and sialic acid. In each lipoprotein, triacylglycerol is a major lipid component (70--83%), followed by phospholipid (8--16%), cholesterol (free and esterified, 8--10%), and free fatty acid (3--4%). Phosphatidylcholine and phosphatidylserine account for 68--74% and 16--24% of the phospholipids, respectively. The fatty acid compositions of total lipids from each lipoprotein are quite similar, with a high degree of unsaturation (63--65%). The carbohydrate content of apolipoprotiens from each lipoprotein is remarkably high (27--31% of apo-lipoproteins) and their composition is very simple: mannose and glucosamine are major constituents in the polysaccharide moiety of each lipoprotein and sialic acid is all in the N-glycolyl form. The amino acid compositions of apo-lipoproteins are quite similar in YLP-1, 2, and 3, with high contents of aspartic acid, glutamic acid, threonine, serine, and leucine. Furthermore, a small amount of glycolipids is present in the yolk lipoproteins. They were separated into six components on TLC. All of them are resorcinol-positive, indicating the presence of sialoglycolipids.

Amino Acids↗

Interspecies comparison of muscle gangliosides by two-dimensional thin-layer chromatography.

Skeletal muscle gangliosides prepared from ten species of animals (human, monkey, bovine, hog, dog, cat, rabbit, guinea pig, mouse, and chicken) were studied by two-dimensional thin-layer chromatography. Densitometric quantification of spots on the chromatograms was carried out with an image analysis system equipped with a computer system. In about thirty-five resorcinol-positive spots, sixteen components could be identified by comparison with authentic standards we had previously obtained. GM3 ganglioside was found as a major component in all examined animals, but there were remarkable species variations in the minor gangliosides. In bovine and hog muscle gangliosides, more than fifteen minor components were detected owing to the presence of N-glycolyl neuraminic acid species. In contrast to this, muscle gangliosides of human, dog, cat, rabbit, and chicken showed relatively simple patterns because of the absence or the small amounts of N-glycolyl type gangliosides. The presence of lactosamine containing gangliosides was characteristic of muscle gangliosides of the examined species.

Animals↗

Identification of four amino acid residues essential for catalysis in human cytidine deaminase by site-directed mutagenesis and chemical modifications.

By site-directed mutagenesis on human cytidine deaminase (CDA), five mutant proteins were obtained: C65A, C99A, C102A, E67D and E67Q. The three cysteine mutants were completely inactive, whereas E67D and E67Q showed a specific activity about 200- and 200000-fold lower, respectively, than the wild-type CDA. Zinc analysis revealed that only E67D, E67Q and C65A contained 1 mol Zn2+/mol subunit as in the wild-type CDA. Kinetic measurements with the specific carboxylic group reagent N-ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinoline performed on wild-type CDA suggest that Glu67 is essential for the catalytic process. Furthermore, when both native and denatured CDA was titrated with 5,5'-dithiobis(2-nitrobenzoic acid) six sulfhydryl groups were detected, whereas in the denatured and reduced enzyme nine such groups were found, according to the sequence data. When p-hydroxymercuriphenyl sulfonate was used, nine sulfhydryl groups were detectable and the release of 1 mol of zinc per mole of CDA subunit was revealed by the metal indicator dye 4-(2-pyridylazo)resorcinol. It seems plausible that the limiting step for the maintenance of zinc in the active site is the formation of coordination between Cys99 and Cys102, whereas Cys65 could lead the zinc to the correct position and orientation within the active site.

Amino Acid Substitution↗

Application of RNA interference to root-knot nematode genes encoding esophageal gland proteins.

Plant parasitic nematodes have been, so far, refractory to transformation or mutagenesis. The functional analysis of nematode genes relies on the development of reverse genetic tools adapted to these obligate parasites. Here, we describe the application of RNA interference (RNAi) to the root-knot nematode Meloidogyne incognita for the knock-down of two genes expressed in the subventral esophageal glands of the nematode and potentially involved in parasitism, the calreticulin (Mi-crt) and the polygalacturonase (Mi-pg-1) genes. Incubation in 1% resorcinol for 4 h induced double-stranded RNA uptake through the alimentary track of the nematodes and led to up to 92% depletion of Mi-crt transcripts. Timecourse analysis of the silencing showed different temporal patterns for Mi-crt and Mi-pg-1. The silencing of Mi-crt was optimal 20 h after soaking, whereas the silencing of Mi-pg-1 was optimal 44 h after soaking. For the two genes, the silencing effect was highly time-limited, since no transcript depletion was detectable 68 h after soaking.

Animals↗

Alcohol reduces GM1 ganglioside content in the serum of inbred mouse strains.

BACKGROUND: Endogenous and exogenous gangliosides in the plasma affect physiologic and pathologic processes such as angiogenesis and atherogenesis. However, the genetic and environmental factors that regulate the expression of plasma gangliosides are not well known. As shown in the liver and the brain, profiles of gangliosides in the plasma may be strain-specific and can be altered by intake of alcohol. Therefore, we analyzed serum gangliosides derived from inbred mouse strains with and without alcohol treatment. METHODS: C57BL/6ByJ (B6By) and BALB/cJ mice (60-70 days old) were injected with 20% alcohol (1-6 g/kg) or saline intraperitoneally, and the ganglioside content of the serum, liver, and cerebellum was measured 4 hr after the injection. Also, the effect of oral alcohol self-administration for 18 days with escalating (3-12%) concentrations of alcohol on the serum GM1 content was studied in B6By mice. The quantification of GM1 was performed with a thin-layer chromatography-staining procedure using a cholera toxin B subunit, and the content of other gangliosides was measured after staining with resorcinol reagent. RESULTS: We found that basal GM1 (containing N-glycolylneuraminic acid) content in the serum of BALB/cJ mice (4.8 +/- 0.26 ng/microl) was 25 times higher than that of B6By mice (0.19 +/- 0.01 ng/microl); the major ganglioside in both strains was GM2. The ganglioside profile in the liver was similar to that of the serum, and the GM1 content in BALB/cJ was nine times higher than that of B6By. Both injection and oral self-administration of alcohol lowered GM1 levels in the serum. CONCLUSIONS: Endogenous ganglioside profiles in the serum are under genetic control among inbred mouse strains, and they can be altered by acute and chronic alcohol administration. These genetic and alcohol-induced differences in the plasma gangliosides, which appear to reflect ganglioside metabolism in the liver, may affect alcohol-related behaviors and pathologic processes.

Animals↗

Relationships between molecular structure and physical properties in bent-core mesogens.

New five-ring bent-core mesogens that possess only ester connecting groups between the aromatic rings and different lateral substituents at the central phenyl ring are presented. The mesophases have been assigned by polarizing microscopy, differential scanning calorimetry, X-ray diffraction and electro-optical measurements. It is shown that the mesophase behaviour depends strongly on the position of the lateral substituents. Compounds, which are derived from 4-cyano-, 4-chloro- and 4,6-dichloro-resorcinol, show polymorphism variants where polar phases (SmAP, SmCP) occur together with nematic and conventional smectic phases, e.g. SmA-SmAP, SmA-SmC(S)P(A)-Col(ob)-SmC(S)P(A), N-SmA-SmCP(A), SmA-SmC-SmCP(A) and SmC-SmCP(A). On the basis of the behaviour of two series of materials, the occurrence of different polar-switching mechanisms could be demonstrated. Apart from the usual mechanism by director rotation around the tilt cone, the polar switching can also take place through collective rotation of the molecules around their long axes, which corresponds to a field-induced switching of the layer chirality. A remarkable finding is the polar switching in the crystalline modification of long-chain, bent-core compounds with a methyl group in 2-position, which is accompanied by a clear change of the optical texture and by a relatively high switching polarization (approx. 600 nC cm(-2)). It was found for selected bent-core compounds that, above the transition temperature of a polar to a non-polar phase, the non-polar phase can be transformed to the polar phase by application of an electric field, which was proved for the transitions isotropic-SmCP(F), SmA-SmCP(F) and isotropic-CrII polar.

Journal Article↗

Surface localization of sialic acid on Actinomyces viscosus.

This study reports the presence of sialic acid in Actinomyces viscosus strains T14V and T14AV. Mild acid hydrolysis of whole organisms released a compound which reacted positively in the periodate-thiobarbituric acid, direct Ehrlich's and resorcinol assays, and which co-chromatographed on paper with authentic N-acetylneuraminic acid. Strain T14V contained 10-fold greater concentrations of sialic acid than did strain T14AV. Sialic acid content was dependent upon the stage of growth of the culture, reaching a maximum in early stationary phase. Epifluorescence microscopy of fluorescein isothiocyanate (FITC)-conjugated Limulus polyphemus agglutinin (LPA), a lectin specific for sialic acid, revealed a uniform distribution of bound lectin on the surfaces of strains T14V and T14AV. Additional evidence for surface localization was obtained by demonstration of whole-cell agglutination of both strains with LPA. All LPA interactions with A. viscosus were inhibited by the presence of 0.1 M-N-acetylneuraminic acid. Neuraminidases from Clostridium perfringens, Arthrobacter ureafaciens and Vibrio cholerae did not release detectable amounts of sialic acid, but the extracellular enzyme from A. viscosus cleaved amounts equivalent to those obtained by acid hydrolysis. Other laboratory strains (W1053, M100, W859, 5-5S, RC45, ATCC 19246, and 'binder') as well as recent clinical isolates of A. viscosus were agglutinated by LPA and released sialic acid upon mild acid hydrolysis. Surface-available sialic acid has been implicated in the inhibition of alternative complement pathway activation and subsequent opsonophagocytosis. Thus the occurrence of surface sialic acid in A. viscosus may represent a mechanism of pathogenesis for this oral bacterium.

Actinomyces↗