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[Change in activity of enzymes for purine metabolism and RNA and DNA biosynthesis in macrophages, reflecting impairment of their functions in neoplastic growth].

The changes in the biochemical parameters of peritoneal macrophages and their coupling to the secretory and phagocytic functions in CH3A mice during the growth of the reinoculated solid hepatoma 22a were studied. The DNA and RNA synthesis during the active tumour growth was more intense than that in resident macrophages. The activity of uridine kinase increased up to 156.0 +/- 12.0 nmol/hour/10(8) but was absent in resident macrophages. This was accompanied by a 7.2-fold increase of interleukin-1 synthesis as determined by the [3H]thymidine incorporation into thymocyte DNA in response to concanavalin A administration to C3H mice. Similar changes were observed in peptone-stimulated macrophages. A specific feature of macrophages from tumour-bearing mice was the impairment of activity of purine exchange enzymes and the efficiency of phagocytosis that were unobserved in peptone-stimulated macrophages. The activity of adenosine deaminase and purine nucleoside phosphorylase was inhibited as a result of their preincubation with zymosan, a phagocytosis-stimulating agent. This was accompanied by a significant decrease of the first chemiluminescence peak resulting from disturbances in Fc-reception. Macrophages of tumour-bearing animals possessed an increased 2.2-fold activity of membrane-bound AMP 5'-nucleotidase concomitant with the lack or decrease of the amplitude of the second chemiluminescence peak reflecting the disturbances in digestion resulting from phagocytosis.

5'-Nucleotidase↗

[Comparative study of 4 methods for isolating Salmonella from surface waters].

The study presented here is the first stage of a project designed to determine the presence of Salmonella serotypes in Lujan river waters, which flow beside the urban area of the homonymous city, in the province of Buenos Aires. It consisted in a comparative study of four methods for Salmonellae detection in 200 samples: Tetrathionate-brilliant green; mannitol-selenite; Rappaport 25 and modified Rappaport-Vassiliadis broth, each of them with and without pre-enrichment in buffered peptone water. The submerged gauze technique was applied to the sampling in three different spots of the river: at the town center, two km water above, and two down-stream from the city. Best results were obtained when modified Rappaport-Vassiliadis enrichment broth and brilliant green agar with 0.25% of sodium deoxycholate were used, after pre-enrichment of the samples in buffered peptone water. Salmonella were isolated from 46% of the samples studied, and 16 serotypes were identified: S. Typhimurium, S. Enteritidis, S. Infantis, S. Mbandaka, S. Israel and S. Subspecies IV 18 z36: z38:-, among them.

Bacteriological Techniques↗

Fatty acid composition of lipopolysaccharides of the strains of different species of Yersinia.

The fatty acid composition of lipopolysaccharides of the strains of Y. enterocolitica, Y. intermedia, Y. frederiksenii and Y. ruckeri studied during cultivation on meat-peptone agar is characterized by the predominance of 3-hydroxytetradecanoic and dodecanoic acids. Closely related to the mentioned bacteria is the strain of Y. kristensenii which is distinguished only by its higher level of hexadecanoic acid. The strains of Y. pseudotuberculosis and the vaccine strain of Y. pestis have a uniform fatty acid composition of lipopolysaccharides with predominance of 3-hydroxytetradecanoic acid. Their relatively low level of dodecanoic acid conditions the characteristic fatty acid spectrum of lipopolysaccharides which differs from that of the above mentioned group of Yersinia. The peculiarities of the fatty acid composition of lipopolysaccharides of both groups of Yersinia are preserved during growth on meat-peptone broth, but the increase in the level of hexadecanoic acid balances the differences between Y. kristensenii, the other Y. enterocolitica-like bacteria and Y. ruckeri. The obtained results confirm close relationship of Y. pseudotuberculosis and Y. pestis, and also of Y. enterocolitica and Y. enterocolitica-like bacteria, showing propinquity of Y. ruckeri to the latter.

Fatty Acids↗

[The fatty acid composition of Haemophilus and Pasteurella multocida cells as a phylogenetic marker].

When grown on meat-peptone agar with heated blood, different Haemophilus species (H. influenzae, H. parahaemolyticus, H. parasuis, H. pleuropneumoniae), including different H. influenzae serovars (a, b, c, d, e, f), and Pasteurella multocida have identical fatty acid composition, characterized by the prevalence of fatty acids with 16 carbon atoms, constituting about 70% and more of the total number of fatty acids, and a low level of fatty acids with 18 carbon atoms. P. multocida strains cultivated on meat-peptone agar with unheated blood have a greatly increased content of fatty acids with 18 carbon atoms, while the content of fatty acids with 16 carbon atoms is much lower. The identity of fatty acid composition under similar cultivation conditions, together with their similarity in other phenetic signs, is indicative of close phylogenic relationship between bacteria belonging to the genus Haemophilus and P. multocida.

Fatty Acids↗

Functional alterations of macrophages in autoimmune MRL-lpr/lpr mice.

To assess the role of macrophages (MAC) in the pathogenesis of systemic lupus erythematosus, we investigated functional aspects of peritoneal MAC obtained from autoimmune MRL/MpJ-lpr/lpr (MRL-lpr) mice. MRL-lpr and control C3H/HeN MAC were obtained from untreated mice or mice injected i.p. with 1 ml of 10% sterile peptone 3 days before cell harvest. MRL-lpr mice had significantly more peritoneal cells (MAC and lymphocytes) than did control mice. In endotoxin-free conditions, MRL-lpr MAC were similar to C3H/HeN MAC in their baseline, and IFN-gamma and/or LPS enhanced cytolysis of 3T12 fibrosarcoma tumor cells. Compared with C3H/HeN MAC, MRL-lpr MAC had a significant increase in antibody-dependent cellular cytotoxicity activity against sheep erythrocytes. This enhanced activity was not accompanied by a similar increase in adherence and/or phagocytosis of the same targets. Finally, in response to phorbol myristate acetate stimulation, both resident and peptone-induced MAC from MRL-lpr mice produced significantly more hydrogen peroxide than did those from control mice. These results indicate that MAC from MRL-lpr mice display features of selective "activation", and suggest that MAC or their products may play a role in the pathogenesis of inflammatory disorders seen in autoimmune diseases.

Animals↗

[Capsule formation in saprophytic yeasts].

Among ten carbon sources and five nitrogen sources tested, D-mannose (3%) and peptone (0.2%) were found to be the best ones for stimulating capsule formation by the yeast Cryptococcus magnus. Addition of yeast extract (0.1%) to the medium favoured formation of larger capsules. Elevated concentrations of mannose and peptone inhibited capsule formation. Comparison of the results obtained with the data about the effect of cultivation conditions on capsule formation by the pathogenic yeast revealed no considerable differences between the latter and the saprophytic yeast.

Carbohydrate Metabolism↗

Properties of Ulex europaeus II lectin isolated by affinity chromatography.

A lectin was isolated from Ulex europaeus seeds by affinity chromatography on affinity adsorbent prepared by copolymerization of acrylamide, N,N'-methylene bisacrylamide and maleylated hog stomach peptone. The lectin is homogeneous as judged by ultracentrifugation (s20,w = 6.4 S), electrophoretic and gel chromatography criteria; it contains 4.2% neutral sugar and 1.4% glucosamine. Its molecular weight is approx. 110,000 and the molecule consists of two noncovalently linked protomers which are formed by two covalently bound basic subunits (Mr = 30,000). The preparation contains three isolectins differing in the strength of interaction with specific sugars (cellobiose, N-acetyl-D-glucosamine) under the conditions of affinity electrophoresis. The lectin is non-specific with human ABO blood group system, the agglutination is inhibited by partial chitin hydrolysate, hog stomach peptone and high concentration of cellobiose.

Amino Acids↗

[Study on the rapid, secondary-growth mutants of Staphylococcus epidermidis (author's transl)].

According to colony type, growth rate and development of secondary growth on the proteose-peptone No. 3 mannitol salt agar (PMS) and the nutrient agar (NA) media, Staphylococcus epidermidis may be classified into three groups. Group I includes strains which develop smooth colonies on both media. Group II consists of those which show rapid propagation of entire clones and develop secondary growth on the PMS medium, but grow only smooth colonies on the NA medium, and which may be called reversible mutants. Group III includes thoes which show secondary growth on both PMS and NA as well as other media, which may be called irreversible mutants. One percent proteose-peptone No. 3 and 5--7% NaCl are the essential ingredients for the induction of mutation, and mannitol can enhance it. Except the high sensitivity of the reversible mutants of human origin, the three groups of chicken origin showed similar drug susceptibility to biosynthesis inhibitors of protein and cell wall. On the HI medium, chloramphenicol inhibited secondary growth of irreversible mutants at 25.0 microgram/ml minimal antimutagenesis concentration (MAC), whereas streptomycin, penicillin, erythromycin and oxytetracycline did not at all. The irreversible mutants had higher resistance to biosynthesis inhibitors of DNA or RNA, e.g. mitomycin C (MMC), novobiocin (NOV) and rifampicin (RIF), than the other two groups. On the HI medium, MMC at the MAC of 0.16 microgram/ml, NA at 25.0 microgram/ml and NOV at 2.5 microgram/ml inhibited the secondary growth of irreversible mutants, but RIF did not. To the irreversible mutants, the MIC and MAC of NA on the PMS medium were both higher than those on the HI medium. The MACs of MMC and NOV on the PMS medium were also higher than those on the HI medium, but their geometric mean MIC remained almost unchanged on both media. Because the MACs of MMC (0.31 microgram/ml) and NA (100.0 microgram/ml) to the reversible mutants on the PMS medium were much similar to those of the irreversible mutants, it suggests that both groups had the similar mutation mechanism.

Anti-Bacterial Agents↗

Trichophyton fischeri sp. nov.: a saprophyte resembling Trichophyton rubrum.

A new species Trichophyton fischeri was isolated as a contaminant on blood agar plates. This fungus is believed to be a saprophyte. It may be confused with T. rubrum. On peptone dextrose agar plate, the growth is white and velvety to cottony. It occasionally forms furrows. The underside of the mature colony is brownish red. Clavate microaleuriospores are common. Trichophyton-type macroaleuriospores are produced occasionally on blood agar and potato dextrose agar. Erythritol does not stimulate T. fischeri to produce a red color on casamino erythritol albumen agar. Closterospore-like projections may be produced on the main filaments on peptone dextrose and potato dextrose agar.

Trichophyton↗

Regulation of porphyrin biosynthesis in yeast. Level of delta-aminolevulinic acid in porphyrin mutants of Saccharomyces cerevisiae.

Saccharomyces cerevisiae mutants bearing mutations at the cyc4 locus are partially deficient in cytochrome synthesis. Although the mutation is not in the structural gene for delta-aminolevulinic acid (Alv) synthase, the mutants are deficient in Alv synthesis in vivo as indicated by abnormally low intracellular Alv concentrations. The cyc4 mutation causes cells to grow very slowly in minimal glucose medium, but not in yeast extract-peptone-glucose medium. A simple nutritional defect caused by the cyc4 mutation is not involved because cytochrome deficiency is enhanced by growing cyc4 cells in yeast extract-peptone medium. A regulatory role for CYC4 is indicated. Evidence for negative feed-back control of Alv synthase by heme is provided by the observation of enhanced intracellular Alv accumulation in yeast mutants partially deficient in decarboxylation of uroporphyrinogen and coproporphyrinogen, respectively.

5-Aminolevulinate Synthetase↗

[Stability of Aujezky's disease virus strain MK25 in an aerosol].

Twenty-two tests were carried out to evaluate the stability of the virus of Aujeszky's disease in aerosol. Use was made of a aerosol chamber of a flow dynamic type, 450 l, of 60 l/min of passing air, 5 min. exposure, 80-100 per cent relative humidity, and temperature range of 14-21.2 degrees C. The size of aerosol particles surpassed 5 micron, and changed according to the composition of the dilution medium of the vaccine. The aerosol was sampled via impinger at the rate of air flow of 5.2 l/min. Tested were the stabilizing properties of 2% glycerin, 10% pepton, 0.8-2.0% saccharose, 2% gelatin, and 5% skimmed milk added to the extender consisting of a 0.2 M phosphate buffer saline, pH 7, specifically for the MK25 strain of the Aujeszky's disease virus in a dispersed status. Highest stability, reaching 11.2% survival rate of the virus in aerosol was recorded at the dispersion of the vaccine in a 0.2 M buffer saline with the addition of 10% pepton and 0.8% saccharose at pH 7.

Aerosols↗

Competitive growth experiments with related pairs of tartrate-fermenting and tartrate-non-fermenting strains of Salmonella typhimurium: relevance to biotyping studies.

For each of the isomers of tartaric acid, meso- or d- or l-, a pair of strains of Salmonella typhimurium was obtained, the one, a naturally occurring, non-fermenting strain and the other a spontaneous, tartrate-fermenting mutant derived from it. For each of the pairs of strains, competitive mixed cultures were grown from inocula of the tartrate-fermenting and tartrate-non-fermenting strains in peptone medium without or with the appropriate tartrate isomer. In each experiment, small numbers of tartrate-fermenting bacteria outgrew small or large numbers of tartrate-non-fermenting bacteria in 24 hours in tartrate-containing but not in tartrate-free peptone medium. The results of the experiments are discussed with reference to the choice of the definitive time of reading for tartrate-utilisation tests in the biotyping of S. typhimurium.

Animals↗

Survival of Campylobacter jejuni in different media and faeces at different temperatures and times of preservation.

Nineteen Campylobacter jejuni strains survived best in suspensions with approximately 10(9) bacteria in 4% albumin solution at-60 degrees C for 17 months (18 out of 19). At the same temperature and during the same period they survived less well in milk (11 out of 19). Recovery of campylobacters after 7 weeks was abundant in albumin, and somewhat less in milk. At -20 degrees C survival in milk was worse; 17 out of 19 cultures did not survive 5 weeks in milk and 7 died in albumin. At 4 degrees C all but one strain survived two weeks in milk and none died in albumin, but after 3 weeks only 7 survived in milk and 2 in albumin. At room temperature campylobacters did not survive 10 days in any of the media used. Peptone water (4%) and peptone-saline proved unsuitable for preservation. From 20 positive stool samples kept at 4 degrees C, on the day after the first sampling C. jejuni could not be recovered from 10 samples; bacteria survived for 2-7 days in 8 samples, for 12-20 days in 2 samples.

Bacteriological Techniques↗

[An optimal blood culture technic].

For blood cultures in septicemic patients it is advisable to use two broth culture bottles. One bottle should be incubated with air, the other without (for anaerobes). Supplemented Peptone Broth II (Becton-Dickinson) and BHI Roche were superior for culturing aerobic and anaerob bacteria in comparison to other blood culture media. They are commercially available, and with these media it is not necessary to inoculate two bottles with blood from one patient. Supplemented Peptone Broth II is also suitable very well for the isolation of Campylobacter fetus and has a redoxpotential which is especially favourable for the isolation of anaerobes.

Bacteria, Aerobic↗

A comparison of methods for the isolation of salmonellae from sewage sludge.

Methods for the isolation of salmonellae from sewage sludge were compared. Buffered peptone water and lactose broth were compared to determine their efficiencies as preenrichment media and temperature and duration of incubation were also investigated. In addition five enrichment and five plating media were compared together with the effects of multiple plating of enrichment broths. Buffered peptone water incubated at 43 degrees C for 24 h was shown to be the pre-enrichment method of choice with enrichment in the RB 10 form of Rappaport's broth incubated at 43 degrees C for 48 h and plating at 24 and 48 h onto brilliant green agar containing sulphamandelate supplement and Hynes' modification of desoxycholate citrate agar. Of the 100 samples used in the study. 96 were found to be positive for salmonella by at least one of the procedures used. A total of 15 Salmonella serotypes were isolated. Salmonella virchow being the most common.

Bacteriological Techniques↗

[Improved method for isolating Yersinia enterocolitica].

Investigations were carried out to restore and isolate Yersinia enterocolitica from both pure and mixed cultures as well as from sewage waters through enriching and alkaline treatment, employing the method of Aulisio and coll. It was found that the peptone used for the purpose (containing sorbitol, 10.0, yeast extract, 5.0, sodium taurocholate, 2.0, bacto-peptone, 2.0, bacto-tryptone, 2.0, bromthymol blue, 0.007, and 1/15 phosphate buffer, 1000 cm3 at pH = 7) made it possible for Y. enterocolitica to replicate at 4 degrees C in 8 to 10 days and inhibited the development of many other organisms. Enriching the medium at 4 degrees C with the alkaline treatment that followed enabled the isolation of Y. enterocolitica from mixed cultures and sewage waters from milk-processing enterprises in a very high percent of the cases as against enriching at 22 degrees C. Such enriching broth could be used not only for the enriching of Y. enterocolitica at 4 degrees C, but also for the differentiation of pure cultures in culturing at the same temperature.

Bacteriological Techniques↗

Relationship between bactericidal and phagocytic activities of peritoneal macrophages induced by irritants.

Cellular accumulation to the peritoneal cavity and modification of various functions of peritoneal macrophages were observed in mice injected intraperitoneally (ip) with thioglycollate medium (TG), liquid paraffin, proteose peptone and Corynebacterium parvum. The cellular composition of peritoneal exudates at 4 days after injection of irritants was almost the same in all the groups and the proportion of macrophages was increased approximately 4 times more than nontreated controls. The ability to kill Listeria monocytogenes and to generate chemiluminescence (CL) were augmented strongly in C. parvum-induced macrophages, while depressed in TG-induced macrophages. The activities of liquid paraffin- or proteose peptone-induced macrophages were almost the same as those in nontreated controls. However, the ability to phagocytose native sheep erythrocytes was greatly augmented both in C. parvum- and TG-induced macrophages. There is thus a discrepancy between bactericidal activity and phagocytic activity among macrophages induced with various irritants.

Animals↗

[Intragastric utilization of antacids following meals in relation to stomach emptying].

Gastric acid secretion and gastric emptying rate was measured using double marker method and continuous titration of a liquid peptone test meal. Titration rate was significantly reduced by 30 ml of an aluminiumhydroxide- and magnesiumhydroxide containing antacid compound (Maalox). Acidity of gastric contents was reduced over a period of 48.4 +/- 9.1 min (mean +/- SD; endpoint of titration pH 5.5) and 77.6 +/- 2.0 min (pH 3.5) (p less than 0.05). The histamine H2-receptor blocker Ranitidine (0.25 mg/kg b.w.) and the antimuscarinic agent Pirenzepine reduced titrable gastric acid secretion in a similar range, as far as the observation period of 90 min is concerned. Biosorbin MCT, a formula diet, stimulated gastric acid secretion half the amount of gastric acid secretion stimulated by the peptone meal. Gastric emptying rate was significantly reduced by formula diet, but not by either of the other compounds.

Adult↗