J Appl Bacteriol, 1989 Jun, 66(6), 469 - 75 Fate of salmonellas in the alimentary tract of chicks pre-treated with a mature caecal microflora to increase colonization resistance; Impey CS et al.; Oral challenge with a strain of Salmonella kedougou, resistant to nalidixic acid, gave a time-course for salmonella survival in the alimentary tract of chicks pre-treated with a caecal culture from an eight-week-old bird to increase colonization resistance . In untreated, control chicks, salmonella colonization of the caeca resulted in counts of greater than 10(6)/g within 48 h and a mean generation time of 1.6 h . With treated birds, however, the salmonellas failed to multiply in the caeca and decreased to a low level over a 48-h period, thus suggesting a mainly bacteriostatic effect . Pre-treatment of chicks with the caecal culture also reduced the proportion of salmonella-positive crop samples . Growth of the salmonella in a feed-slurry system resembling moistened crop contents occurred rapidly at 37 degrees C but was prevented by addition of the treatment culture, a decline in numbers coinciding with Lactobacillus populations of ca 10(9)/g and a pH value of 5.5 . Incorporation of a feed decontaminant, 1% formic acid (pH 4.0), rapidly eliminated both salmonellas and organisms added from the treatment culture. Eur Respir J, 1989 Jun, 2(6), 589 - 91 Lactobacillus pneumonia in a patient with oesophageal carcinoma; Querol JM et al.; We report a case of necrotizing pneumonia caused by Lactobacillus secondary to a tracheo-oesophageal fistula created by an oesophageal carcinoma . We emphasize the presence of resistance of Lactobacillus to clindamycin and cotrimoxazole, previously reported to be effective. Scand J Dent Res, 1989 Jun, 97(3), 257 - 62 Effect of sorbitol- and xylitol-containing chewing gum on salivary microflora, saliva, and oral sugar clearance; Wennerholm K et al.; The effect of frequent use of a sorbitol-containing nicotine chewing gum on saliva secretion rate and buffer capacity and some oral bacteria was studied in 27 patients at a smoking cessation clinic . The effect was compared with that obtained after frequent use of a chewing gum containing xylitol in a second study in 14 subjects . The results showed that sorbitol-containing nicotine chewing gum had no significant effect on salivary numbers of oral streptococci and lactobacilli during a 3-month period of active chewing five times a day . Chewing on xylitol-containing gum caused a significant decrease in salivary S . mutans after 2 months but not after 3 months . No change in secretion rate or buffer capacity was observed in the two studies . Oral sugar clearance time was reduced after 3 months with a statistically significant difference to baseline values in subjects consuming the sorbitol-containing nicotine chewing gum. Antibiot Khimioter, 1989 Jun, 34(6), 430 - 2 {Reciprocal effect of Klebsiella pneumoniae and Lactobacillus on their cytadhesion}; Lentsner AA et al.; Reciprocal effect of 4 strains of Klebsiella pneumoniae and 6 Lactobacillus strains on their cytadhesion in mixed populations was studied on a model of formalinized human erythrocytes . The Lactobacillus strains included 2 strains of Lactobacillus casei subsp . casei, 2 strains of L . plantarum and 2 strains of L . fermentum . It was shown that adhesion of both the Klebsiella and the Lactobacillus strains changed under their reciprocal effect . The changes were characterized by the strain differences and depended on the quantitative ratio of the microorganisms. Acta Odontol Scand, 1989 Jun, 47(3), 175 - 83 Some salivary factors in insulin-dependent diabetics; Thorstensson H et al.; The aim of the study was to compare salivary flow rate, salivary pH, buffer capacity of the saliva, salivary glucose content, and number of Candida albicans, lactobacilli, and Streptococcus mutans in the saliva in age- and sex-matched adult long- and short-duration insulin-dependent diabetics and non-diabetics . Ninety-four long-duration and 86 short-duration diabetics and 86 non-diabetics, aged 20-70 years, participated in the study . Paraffin-stimulated whole saliva was collected . Both long- and short-duration diabetics had a decreased salivary flow rate and an increased salivary glucose content compared with non-diabetics . However, the differences were small . There were no significant differences between the groups in salivary pH, buffer capacity, or bacterial counts. Aichi Gakuin Daigaku Shigakkai Shi, 1989 Jun, 27(2), 495 - 509 {Salivary microbial flora of mentally retarded persons with rumination}; Ishiguro H; The salivary microflora of institutionalized mentally retarded persons with chronic rumination was investigated, especially emphasizing the occurrence of the aciduric microorganisms such as mutans streptococci, lactobacilli and yeasts . 10 of those subjects together with 10 control subjects resident in the same institution were selected at random . The control subjects were mentally retarded but without rumination . Samples of saliva were collected about 2 hours after lunch . The results obtained were as follows: 1 . Dental caries was more prevalent and advanced in those ruminating than the control subjects . 2 . The mean salivary pH-value in the ruminating subjects was significantly lower than the control subjects . 3 . The proportional distribution of the genera or species of salivary microorganisms cultivated on the blood agar plates was similar for both the ruminating and non-ruminating subjects . The only difference was observed in the genus of Actinomyces, which was less frequently isolated from the ruminating subjects . 4 . Mutans streptococci, lactobacilli and yeasts recovered from their respective selective media were significantly predominant in those ruminating compared with control subjects . 5 . Positive correlations among the cell count of mutans streptococci, lactobacilli and yeasts were found in the ruminating subjects . The yeasts isolated were classified exclusively into Candida albicans . 6 . The results described above suggest that long-lasting acidic milieu in the oral cavities of the ruminating subjects favors the proliferation of mutans streptococci, lactobacilli and yeasts, resulting in the prevalence of dental caries in those subjects. Oralprophylaxe, 1989 Jun, 11(2), 70 - 3 {Comparison of clinical and microbiological parameters for determination of actual caries risk}; Schlagenhauf U et al.; In 28 children and adolescents, age 4-17, the numbers of decayed tooth surfaces, filled surfaces and the extent of plaque coverage measured with a plaque scoring index were recorded as clinically assessible caries risk factors . Subsequently paraffin stimulated saliva samples were taken from the same subjects and evaluated microbiologically for their content of caries related microorganisms (S . mutans, lactobacilli) . Statistical analysis of correlation revealed only weak, mostly not significant correlations between clinical and microbiological findings . The results are discussed. J Med Chem, 1989 Jun, 32(6), 1284 - 9 Folate analogues . 32 . Synthesis and biological evaluation of 2-desamino-2-methyl-N10-propargyl-5,8-dideazafolic acid and related compounds; Patil SD et al.; The chemical synthesis of three close analogues (2-4) of N10-propargyl-5,8-dideazafolate (PDDF) is described . The quinazoline ring of 2 and 4 was constructed from the pivotal intermediate 9 in a novel and unambiguous manner during the final step of the synthesis under very mild conditions . 2-Desamino-2-methyl-N10-propargyl-5,8-dideazafolate (DMPDDF) (2) was a strong inhibitor of human and Lactobacillus casei thymidylate synthases, whereas 2-desamino-2-(trifluoromethyl)-N10-propargyl-5,8-didezafolate (3) and 2-desamino-2,3-dimethyl-N10-propargyl-5,8-dideazafolate (4) were only weak inhibitors of this enzyme . DMPDDF exhibited excellent growth inhibition of Manca human lymphoid leukemia and H35 hepatoma cells in culture . The inhibitor activities of 2 were 43- and 65-fold greater than that of PDDF, respectively, in these cell lines . H35R cells that are resistant to methotrexate by virtue of a transport defect were cross resistant to DMPDDF but not to PDDF . H35FF cells which have 70-fold greater amounts of thymidylate synthase compared to H35N cells were 130-fold resistant to DMPDDF . Furthermore, the toxicity of DMPDDF to H35 hepatoma cells could be completely reversed by thymidine, establishing its locus of action as thymidylate synthase . Transport studies in vitro established that DMPDDF effectively inhibits MTX influx into H35 hepatoma cells, whereas PDDF has no effect on MTX transport in this cell line . These data suggest that the greater activity of DMPDDF relative to PDDF is partly due to the ability of the former compound to enter cells via the MTX/reduced folate transport system . Enzyme inhibition data of 4 suggest that the presence of N3H in DMPDDF is essential for binding to thymidylate synthase. Oral Microbiol Immunol, 1989 Jun, 4(2), 57 - 64 Competition between Streptococcus mutans and Lactobacillus casei in mixed continuous culture; Bowden GH et al.; In natural oral ecosystems, succession of Streptococcus mutans and Lactobacillus species occurs in bacterial communities which can be associated with the development of carious lesions . The objective of this study was to explore the interaction between 2 strains of S . mutans (SF2452) and Lactobacillus casei (RB1014) when grown together in continuous culture under conditions of varying environmental pH and in the presence of fluoride . When the strains, isolated from approximal carious lesions, were grown initially at a dilution rate of 0.1 h-1 under a glucose limitation at pH 7.0, a stable mixed community developed (Days 5-10) with S . mutans as the dominant population . When the pH control was removed (Day 10), the binary culture lowered the pH to 5.1 causing a small reduction in the numbers of S . mutans and allowing L . casei to predominate . A more dramatic effect was seen when the environmental pH was rapidly lowered to 4.8 by the addition of acid . In this case, the S . mutans population was significantly reduced and the L . casei population rose to the level previously held by S . mutans . Re-adjustment of the pH to 7.0 resulted in a re-establishment of the population ratios prior to the addition of acid . The possibility that S . mutans was capable of aciduric adaptation during a biologically-generated pH reduction was examined by mixing cultures of both organisms after each had been grown to steady state at pH 5.5 in separate chemostats. J Dairy Sci, 1989 Jun, 72(6), 1408 - 17 Genetic transfer systems for delivery of plasmid deoxyribonucleic acid to Lactobacillus acidophilus ADH: conjugation, electroporation, and transduction; Luchansky JB et al.; Lactobacillus acidophilus ADH is a bacteriocin-producing human isolate that adheres to human fetal intestinal cells and human ileal cells . We have employed both electroporation and conjugation methodologies to transfer various plasmids to L . acidophilus ADH . Furthermore, we have demonstrated transduction of plasmid DNA within this strain by a temperate bacteriophage (phi adh) harbored by L . acidophilus ADH . Plasmid pGK12 was introduced into strain ADH by electroporation at frequencies as high as 3.3 X 10(5) transformants/micrograms of plasmid DNA . Transconjugants of strain ADH were recovered at frequencies of 10(-2) (pAMB1), 10(-4) (pVA797::Tn917), and 10(-4) (pVA797) per donor cell after filter-mating with Lactococcus lactis ssp . lactis . Plasmid pGK12 was transduced from a phage phi adh lysogen into a recipient strain of L . acidophilus ADH at an average frequency of 3.4 X 10(-8) transductants/pfu . Transformants, transconjugants, or transductants were verified by both phenotype and plasmid profile for acquisition of plasmid DNA . The ability to transfer plasmids and mobilize DNA sequences by electroporation, conjugation, and transduction will augment our efforts to define and characterize the activities of L . acidophilus in the intestinal tract. Zentralbl Hyg Umweltmed, 1989 Jun, 188(3-4), 294 - 302 Application of non-radioactive BACTEC system to sterility testing and hygiene; Hammann R et al.; The applicability of the non-radioactive BACTEC NR 660 system for the detection of low viable counts of aerobic and anaerobic bacteria (including non-fermenters and aerobic spore-forming bacteria) and fungi was investigated . The BACTEC system together with the usual BACTEC NR 6A and NR 7A media was able to detect most of the organisms without supplementation of the media; for growth of lactobacilli and other acidophilic organisms, sterile orange juice was used as a supplement . Microbial growth was detected from pure cultures and also from artificially contaminated beverages and ointment bases . Detection times varied according to the types of metabolism of the respective strains and their initial viable counts, but usually they were detected after 20 to 42 h of incubation . The BACTEC NR 660 system seems to be applicable not only for blood cultures in medical microbiology but also in pharmaceutical and food industry. J Med Chem, 1989 Jun, 32(6), 1277 - 83 Folate analogues . 31 . Synthesis of the reduced derivatives of 11-deazahomofolic acid, 10-methyl-11-deazahomofolic acid, and their evaluation as inhibitors of glycinamide ribonucleotide formyltransferase; Nair MG et al.; The Boon-Leigh procedure, involving condensation of a 6-chloro-5-nitropyrimidine (22) with an alpha-amino ketone (20 or 21) followed by reduction of the nitro group, cyclization, and L-glutamylation, led to the formation of 11-deazahomofolate (29) and its 10-methyl derivative (30) . The corresponding (6R,S)-5,6,7,8-tetrahydro (4, 5) and 7,8-dihydro (31, 32) derivatives were prepared by catalytic hydrogenation . (6S)-11-Deazatetrahydrohomofolate was prepared from 29 by enzymatic reduction . Compounds 29 and 30 had little effect (IC50 greater than 2 x 10(-5) M) on Lactobacillus casei glycinamide ribonucleotide (GAR) formyltransferase but (6R,S)-11-deazatetrahydrohomofolate (4) is a potent inhibitor of this enzyme (IC50 = 5 x 10(-8) M) . It is at least 100 times more inhibitory than 33, the 6S compound, indicating that the 6R component of the mixture having the unnatural configuration at C6 (34) is responsible for the potent inhibition . Compound 4 is a much weaker inhibitor of murine (L1210) and human (MOLT-4) leukemia cell GAR formyltransferases (IC50 greater than 1 x 10(-5) M) . (6R,S)-11-Deaza-10-methyltetrahydrohomofolate (5) (IC50 = 1.1 x 10(-5) is 200 times weaker than 4 against L . casei GAR formyltransferase . However, 11-deaza-10-methyldihydrohomofolate (32) is more inhibitory (IC50 = 5.5 x 10(-7) M) than 5 or 30 . None of the compounds showed inhibition of L . casei aminoimidazolecarboxamide ribonucleotide (AICAR) formyltransferase, dihydrofolate reductase, or thymidylate synthase . The dihydro derivatives 31 and 32 are 5% as active as dihydrofolate as substrates for L . casei dihydrofolate reductase . Compound 4 showed moderate inhibition of the growth of L . casei, Streptococcus faecium, MOLT-4 cells, and MCF-7 human breast adenocarcinoma cells. Biochem J, 1989 May 15, 260(1), 163 - 9 Reaction of Woodward's reagent K with D-xylose isomerases . Modification of an active site carboxylate residue; Vangrysperre W et al.; D-Xylose isomerases from Streptomyces violaceoruber, Streptomyces sp., Lactobacillus xylosus, Lactobacillus brevis and Bacillus coagulans were rapidly inactivated by Woodward's reagent K . Second-order rate constants in the absence of ligands, at pH 6.0 and 25 degrees C, were 41, 36, 22, 95 and 26 M-1.min-1 respectively . Spectral analysis at 340 nm revealed that inactivation was correlated with modification of five, six, two, three and six carboxylate residues per monomer respectively . In the presence of protecting ligands, modification of one carboxylate group was prevented . The results support the idea of an active site glutamate or aspartate group that may contribute to the catalytic activity of all these D-xylose isomerases. Gene, 1989 May 15, 78(1), 47 - 57 Cloning, sequencing and expression in Escherichia coli of the D-2-hydroxyisocaproate dehydrogenase gene of Lactobacillus casei; Lerch HP et al.; D-2-Hydroxyisocaproic acid dehydrogenase (D-HicDH) from Lactobacillus casei was purified and partially sequenced . A 65-mer oligodeoxyribonucleotide probe corresponding to the N-terminal 23 amino acids was synthesized and a physical map was made of the genomic region which hybridized most strongly . A strongly hybridising restriction fragment was highly purified and eventually cloned at low frequency in pBR322 . The original clones spontaneously produced D-HicDH at about 0.05% of total protein and showed viability problems in that 10- to 12-h growth-lag periods occurred after diluting stationary cultures into fresh medium . Subcloning into pGEM3 plasmids for sequencing with concomitant ExoIII deletion led to clones which no longer exhibited the growth inhibition characteristics but now made D-HicDH as 3 to 5% of total protein . Subcloning downstream from a double PL PR promoter in plasmid pJLA601 gave a highly inducible clone that builds large inclusion bodies of largely denatured D-HicDH . The gene transcript was mapped for L . casei and Escherichia coli hosts . The promoter, terminator and Shine-Dalgarno sequence are functional in both organisms . The gene encodes a protein subunit of 38 kDa, whereby 67% of the sequence could be checked by correlation with partial peptide sequences from the original enzyme . So far no Lactobacillus gene has been found to utilize the Arg codons AGG and AGA. Arch Inst Cardiol Mex, 1989 May-Jun, 59(3), 287 - 92 {Etiology of infectious endocarditis at the Instituto Nacional de Cardiologia Ignacio Chavez from 1976 to 1986}; Reyes Bribiesca R et al.; We studied 85 cases of infectious endocarditis (IE) with an anatomical diagnosis (biopsy or necropsy) . Most cases occur between the second and fourth decade of life; male/female ratio was 1/0.8; 45 had IE in a natural valve; 40 had prosthetic IE . Clinical diagnosis was established or at least suspected in 61 cases (72%); the most common misdiagnosis was acute rheumatic fever . Blood cultures were positive in 25 cases of natural valve IE (56%) and in 25 cases of prosthetic IE (63%) . Half of those negative culture cases had a history of antimicrobial use before their arrival at the hospital . Those microorganisms which cause natural valve IE were: staphylococci (9), streptococci (8), Gram negatives (5), Candida sp (1), and two non-identified cases; 1 patient had a polymicrobial infection . Prosthetic IE was divided into early and late infections . The former was due to: staphylococci (4), streptococci (1), Candida sp (1); the latter: staphylococci (7), streptococci (4), Gram negatives (4), Candida sp, Mycobacterium fortuitum and Peptococcus (1 each), and in a single case Lactobacillus sp, presumably a contaminant. Antimicrob Agents Chemother, 1989 May, 33(5), 674 - 9 Chemical characterization of an antimicrobial substance produced by Lactobacillus reuteri; Talarico TL et al.; Lactobacillus reuteri converts glycerol into a potent cell growth inhibitor . This substance, termed reuterin, inhibits the growth of gram-positive and gram-negative bacteria as well as yeasts, fungi, and protozoa . Semipreparative chromatography was used to purify reuterin, and Fourier transform infrared spectroscopy and liquid chromatography-mass spectrometry were used to establish the molecular weight as well as the molecular functionality of the reuterin molecule . Nuclear magnetic resonance studies of purified reuterin carried out with deuterium oxide confirmed the presence of two three-carbon compounds, beta-hydroxypropionaldehyde and the corresponding hydrated acetal, and a six-carbon cyclic dimer of the aldehyde . Further nuclear magnetic resonance studies with deuterated methanol revealed that in this solvent the compound existed as a three-carbon compound in a methoxy form . Trimethylsilyl derivatives of reuterin were analyzed by gas chromatography-mass spectrometry, and a molecule was identified which had a molecular weight corresponding to a disilylated dimeric structure . On the basis of the above information, reuterin was determined to be an equilibrium mixture of monomeric, hydrated monomeric, and cyclic dimeric forms of beta-hydroxypropionaldehyde . This was subsequently confirmed by chemical synthesis. Infection, 1989 May-Jun, 17(3), 188 - 93 Newly recovered and delineated microbial species of the human genital tract; Hammann R; Several new genitourinary Bacteroides species and the new Mobiluncus/Falcivibrio have been described since 1977 . The normal vaginal flora consists mainly of lactobacilli and other "non-pathogenic" aerobes and anaerobes . Under unknown circumstances, organisms may increase in number, displacing the normal lactobacillus flora and raising the vaginal pH . This renders a more favourable biotope for organisms with a pH optimum in the neutral or alkaline range . Overgrowth of Gardnerella, Bacteroides, B-streptococci and Mobiluncus/Falcivibrio is indicative of vaginosis, which is not a typical sexual disease, but may be sexually transmitted . Proper treatment includes the male partner . No overall-valid therapy exists, but metronidazole is the drug of choice if Bacteroides or Gardnerella form the disturbed flora, and beta-lactams are the drugs of choice when Mobiluncus or aerobes are involved. J Gen Microbiol, 1989 May, 135 ( Pt 5), 1175 - 86 Protein-mediated adhesion of Lactobacillus fermentum strain 737 to mouse stomach squamous epithelium; Conway PL et al.; The mechanism of adhesion of Lactobacillus fermentum strain 737 to mouse stomach squamous epithelium was investigated . Adhesion inhibition tests involving chelators, monosaccharides, periodate and concanavalin A and the use of bacteria grown in the presence of tunicamycin failed to clarify the adhesive mechanism . Washed bacterial cells had reduced adhesive capacity, except in the presence of spent broth culture supernatant fraction or cell washings . Spent culture supernatant fractions of erythrosine-supplemented broth did not enhance adhesion of washed cells . The adhesion-promoting factor(s) in the spent broth culture supernatant fractions and cell washings bound to both bacterial and epithelial cell surfaces, but did not promote adhesion of two other Lactobacillus strains which were not of mouse origin, thereby indicating host specificity for the adhesion-promoting activity . Chemical characteristics of the adhesion-promoting factor were determined by pretreatment of the dialysis retentate of spent broth culture supernatant fractions with proteolytic enzymes, concanavalin A-Sepharose or periodate before the adhesion assay . The adhesin was non-dialysable, pronase-sensitive, heat sensitive at 100 degrees C, had no affinity for concanavalin A-Sepharose and contained no carbohydrate groups active in the adhesion process . The protein profiles of dialysis retentates of spent broth culture supernatant fractions after bacterial growth in the absence and presence of erythrosine were determined by 2-dimensional SDS-PAGE . Gel filtration by HPLC was used for purification of an adhesion-promoting fraction . The host-specific adhesion of L . fermentum strain 737 was mediated by a protein, with an Mr of 12-13000, that was not detectable in cells grown in the presence of erythrosine . A model for the mode of binding of the adhesin to host epithelia and bacterial surfaces is proposed. J Gen Microbiol, 1989 May, 135 ( Pt 5), 1167 - 73 Role of erythrosine in the inhibition of adhesion of Lactobacillus fermentum strain 737 to mouse stomach tissue; Conway PL et al.; The mechanism by which the food colour erythrosine inhibits the adhesion of Lactobacillus sp . to squamous epithelium in the mouse stomach was investigated using an in vitro adhesion assay . Inhibition of adhesion occurred only after growth of L . fermentum in erythrosine which bound to the bacterial cell surface . Erythrosine did not interfere with the receptor on the epithelial cell surface . Growth, but not the ATP content per cell, was affected by the presence of erythrosine in the growth medium . No consistent correlation between hydrophobicity and growth in two different broths was noted when erythrosine was present . Analyses of phenol/water extracts and transmission electron micrographs revealed no reduction in extracellular polysaccharide after growth in the presence of erythrosine . It was concluded that erythrosine affects bacterial metabolism thereby preventing production of the bacterial adhesin which is not the extracellular polysaccharide. Biochimie, 1989 May, 71(5), 639 - 44 Presence of an L(+)-lactate dehydrogenase in cells of Lactobacillus delbrueckii ssp . bulgaricus; Ragout A et al.; Lactobacillus delbrueckii ssp . bulgaricus ATCC 11842 was cultured in a chemostat and growth conditions were varied as required . Synthesis of L(+)-lactate was observed in all cases as well as activity of L(+)-lactate dehydrogenase in cell-free extracts . This enzyme was responsible for the formation of the L(+) isomer of lactate, since a lactate racemase was not present. Thorac Cardiovasc Surg, 1989 Apr, 37(2), 99 - 102 Recurrent infective endocarditis in idiopathic hypertrophic subaortic stenosis; Stulz P et al.; A patient with idiopathic hypertrophic subaortic stenosis suffered from five episodes of infective endocarditis of the mitral valve . At least two of these episodes were caused by Lactobacillus species, a microorganism known to be difficult to eradicate . Definitive cure was only achieved by surgical treatment. J Clin Microbiol, 1989 Apr, 27(4), 724 - 30 Identification of gram-positive coccal and coccobacillary vancomycin-resistant bacteria; Facklam R et al.; A total of 84 of 150 vancomycin-resistant (defined as no inhibition of bacterial growth around a 30-micrograms vancomycin disk placed on 5% sheep blood-Trypticase soy agar {BBL Microbiology Systems, Cockeysville, Md.}) bacteria were definitively identified by determining the phenotypic criteria . The identity of representatives was also confirmed by DNA-DNA hybridizations . The following strains were identified: 1 Enterococcus faecium, 18 Leuconostoc mesenteroides, 15 Leuconostoc citreum, 9 Leuconostoc pseudomesenteroides, 2 Leuconostoc lactis, 20 Pediococcus acidilactici, 5 Pediococcus pentosaceus, and 14 Lactobacillus confusus . The remaining vancomycin-resistant strains were identified as probable Leuconostoc (6 strains), probable Pediococcus (1 strain), and probable lactobacilli (28 strains) . A total of 32 strains of gram-positive coccobacillary bacteria remained unidentified . Tests used for the phenotypic identification of strains to the genus level included a Gram stain of bacteria grown in thioglycolate broth, gas production in Lactobacillus Mann, Rogosa, and Sharpe broth, hydrolysis of pyrrolidonyl-beta-naphthylamide, bile-esculin reaction, demonstration of streptococcal group D antigen, and growth at 10 and 45 degrees C and in 6.5% NaCl broth . Strains were identified to the species level by hydrolysis of esculin, reactions in litmus milk, slime production on 5% sucrose agar, acidification of maltose, melibiose, and raffinose broths, deamination of arginine, and growth at 42 degrees C and in 6.5% NaCl broth. Scand J Dent Res, 1989 Apr, 97(2), 171 - 7 Salivary immunoglobulins, lysozyme, pH, and microbial counts in children receiving anti-neoplastic therapy; Pajari U et al.; Salivary immunoglobulins, lysozyme, pH, and microbial counts were determined in 55 children with cancer diseases (37 cured subjects and 18 acute ones) and 103 healthy subjects . 5-10 ml unstimulated whole saliva was collected and pH, immunoglobulins and lysozyme were measured . Chairside dip-slide cultivations were used for microbiologic cultures . Reduced salivary pH and an increased amount of lysozyme were found in the saliva of those children who had been cured of their cancer diseases, but ongoing cancer disease or the treatment provided for it reduced pH and increased the amounts of lysozyme, lactobacillus, Streptococcus mutans and some immunoglobulins . These findings suggest that children with childhood cancer may be more susceptible to dental diseases than healthy ones. Scand J Dent Res, 1989 Apr, 97(2), 150 - 8 Subgingival microflora, dental and periodontal conditions in patients with hereditary fructose intolerance; Saxen L et al.; Hereditary fructose intolerance (HFI) is a rare autosomally recessive disease which leads to severe hypoglycemia . The fructose-free diet of these patients apparently influences dental health . Half of the patients are free of caries, but there are no reports on their periodontal condition nor on the composition of their subgingival microflora . Therefore 18 patient with HFI were examined for the following parameters: radiographic bone loss, caries rate, gingival bleeding, occurrence of calculus, pocket depths, salivary flow rate, buffer capacity and pH from paraffin wax-stimulated saliva . Salivary S . mutans and lactobacilli were also enumerated . Specific antibody titers from whole saliva and serum to Actinobacillus actinomycetemcomitans gamma 4, Bacteroides gingivalis, and Capnocytophaga ochracea were determined with ELISA . Finally the subgingival plaque was analyzed by culture . Altogether 196 anaerobic or microaerophilic isolates representing 29 different species were obtained from the HFI patients and 164 isolates of 26 species from the controls . The frequency of Actinomyces odontolyticus, Veillonella parvula, and Wolinella recta in the HFI patients was significantly high The DMF-induces were lower in the study group than in the controls and so was the occurrence of S . mutans and lactobacilli in the saliva . Of the periodontal parameters examined, the only difference found was an increased incidence of gingival bleeding and calculus formation in the HFI patients . The results suggest that in the HFI patients the common gingival bleeding is associated with the more frequent occurrence of Actinomyces odontolyticus, Veillonella parvula, and Wolinella recta. Scand J Dent Res, 1989 Apr, 97(2), 120 - 5 Distribution of mutans streptococci in populations with different levels of sugar consumption; Carlsson P; The observations in the Vipeholm Study regarding sugar consumption and oral lactobacillus counts were reviewed in relation to recent data on the epidemiology of mutans streptococci . The mutans streptococci were not present in higher counts in a population with high level of sugar consumption compared to two populations with a low level . As expected, differences were found in the caries prevalence between the different populations . An association between bacterial count and dental caries status could be demonstrated at the low level of sugar consumption and caries prevalence . Strains of mutans streptococci isolated in a population with extremely low caries prevalence were shown to be cariogenic in the hamster model . It is apparent that the observations made in the Vipeholm Study and the present finding have similarities, although different bacteria are studied . It was concluded that the difference in diet between the populations or groups were not sufficient to affect the counts of lactobacilli or mutans streptococci, but may have been sufficient to create the differences in caries prevalence. Scand J Dent Res, 1989 Apr, 97(2), 110 - 4 Microbial aspects of frequent intake of products with high sugar concentrations; Carlsson J; The Vipeholm Study showed that a frequent intake of products with high sugar concentrations increases caries activity . This effect of sugar was explained by the acid production of bacteria on the teeth . In the following 40 yr many microbial characteristics have been described, which are important in the pathogenesis of dental caries . This contribution focuses on one characteristic of the cariogenic bacteria; their acidurance . Compared to other oral bacteria the cariogenic lactobacilli and mutans streptococci have enzymes with greater resistance to acid, and greater capacities of the cells to extrude protons from the cytoplasm . The most important enzyme for the extrusion of protons is a membrane-bound proton-translocating ATPase . This enzyme is found in higher amounts and is working at lower pH in the cariogenic bacteria . Another important characteristic of the cariogenic bacteria is their production of lactic acid when they are exposed to high concentrations of sugar . Lactic acid has a pK of 3.8, while other carboxylic acids have a pK of 4.8 . The bacterial cell membrane is permeable to undissociated carboxylic acids . These undissociated acids act as carriers to bring protons back into the cytoplasm . Bacteria with lactic acid as fermentation product have therefore the possibility to grow and produce acids at one pH unit lower than bacteria with, for example, acetic, propionic, or butyric acid as fermentation product. J Anim Sci, 1989 Apr, 67(4), 966 - 74 Anticholesteremic property of Lactobacillus acidophilus yogurt fed to mature boars; Danielson AD et al.; Three strains of Lactobacilus acidophilus (LA) were isolated from the feces of mature boars that were not being fed antibiotics from the Nebraska Gene Pool (NGP) . All three LA isolates were screened in vitro for anticholesteremic and antimicrobial activities . One strain, LA16, caused the greatest reduction in cholesterol and inhibited both Bacillus subtilis and Escherichia coli the most . LA16 was used to produce 16, 18.9-liter quantities of acidophilus yogurt (AY), over a period of 8 wk, for use as a feed ingredient in diets for the NGP boars . Colony forming units (cfu), pH, protein, energy, Ca and P were consistent across all 16 batches of yogurt . All of the 18 boars were fed a high-cholesterol diet for a period of 56 d at a rate of 2.268 kg/(hd.d) to furnish 6.661 g/(hd.d) of cholesterol . Nine of the boars then were fed 1.81 kg/(hd.d) of a second diet that was supplemented with .454 kg/(hd.d) of AY . The other nine boars were fed the original diet . Cholesterol intake was the same for the two dietary treatments . Blood samples were collected weekly from the brachial-jugular region and the sera were analyzed for lipids . Acidophilus yogurt reduced serum cholesterol (P less than .01) and low density lipoproteins (P less than .08), but it had no effect on serum triglycerides (P greater than .23) or on high density lipoproteins (P greater than .11). J Biol Chem, 1989 Mar 15, 264(8), 4722 - 4 Crystallization and molecular symmetry of ornithine decarboxylase from Lactobacillus 30a; Momany C et al.; Ornithine decarboxylase from Lactobacillus 30a is representative of the large subunit (80 kDa), oligomeric, pyridoxal phosphate-dependent amino-acid decarboxylases . Yellow crystals of ornithine decarboxylase are obtained from polyethylene glycol solutions and belong to space group P6 with unit cell constants a = b = 194.9 and c = 97.44 A, alpha = beta = 90 degrees and gamma = 120 degrees, V = 3.21 x 10(6) A3 . Still photographs show reflections at better than 2.4-A resolution . Electron micrographs reported by Guirard and Snell (Guirard, B.M., and Snell, E.E . (1980) J . Biol . Chem . 255, 5960-5964) reveal that the ornithine decarboxylase dodecamer is a hexagonally shaped particle with a point-to-point distance of approximately 210 A and a thickness of approximately 70 A . The crystallographic unit cell can thus accommodate one 10(6)-Da dodecamer (Vm = 3.2 A3/Da), implying that a dimer occupies an asymmetric unit . Tanaka rotation function analysis, using native data (5-7 A) collected from three crystals, reveals that the particle has the expected 622 molecular symmetry with molecular 2-fold axes lying at 20 degrees and 50 degrees from a in the a-b plane . A search for suitable heavy atom derivatives is underway. Biochemistry, 1989 Mar 7, 28(5), 2297 - 305 Dihydrofolate reductase: multiple conformations and alternative modes of substrate binding; Birdsall B et al.; The complex of Lactobacillus casei dihydrofolate reductase with the substrate folate and the coenzyme NADP+ has been shown to exist in solution as a mixture of three slowly interconverting conformations whose proportions are pH-dependent {Birdsall, B., Gronenborn, A . M., Hyde, E . I., Clore, G . M., Roberts, G . C . K., Feeney, J., & Burgen, A . S . V . (1982) Biochemistry 21, 5831} . The assignment of the resonances of all the aromatic protons of the ligand molecules in all three conformational states of the complex has now been completed by using a variety of NMR methods, particularly two-dimensional exchange experiments . The resonances of the nicotinamide protons of the coenzyme and the pteridine 7-proton of the folate have different chemical shifts in the three conformations, in some cases differing by more than 1 ppm . Comparison of the COSY spectra of the complex at low pH (conformation I) and high pH (conformations IIa and IIb) with that of the enzyme-methotrexate-NADP+ complex shows only slight differences in the conformation of the protein . The pattern of chemical shift changes in the ligand and the protein indicates that the structural differences are localized within the active site of the enzyme . Nuclear Overhauser effects (NOEs) are observed between the nicotinamide 5- and 6-protons and the methyl resonance of Thr 45 at both low and high pH, indicating that there is no major movement of the nicotinamide ring . By contrast, NOEs are observed between the pteridine 7-proton and the methyl protons of Leu 19 and Leu 27 in conformations I and IIa but not in conformation IIb.(ABSTRACT TRUNCATED AT 250 WORDS) Oral Surg Oral Med Oral Pathol, 1989 Mar, 67(3), 286 - 90 Salivary and microbial conditions and dental health in patients with Crohn's disease: a 3-year study; Sundh B et al.; Salivary and microbial conditions and dental status were examined in 21 patients randomly selected from a group of patients with Crohn's disease . Three years later, the patients were recalled and the same factors were reexamined . The results showed that the caries experience in these patients was higher than that observed in age-matched subjects in a normal reference population . Dietary examination disclosed no changes in intake frequencies of different common food items during the 3-year period . All patients had a normal saliva flow rate and buffer capacity . Protein concentration tended to increase and sialic acid to decrease when compared with available reference data . The number of salivary mutans streptococci organisms and lactobacilli was high both at baseline and after 3 years, and no significant differences were observed between the two examinations . Patients with Crohn's disease belong to a high-risk group in relation to caries and should be given comprehensive preventive caries measures. Indian J Med Res, 1989 Mar, 89, 80 - 6 Gardnerella vaginalis vaginitis in Cu T 200 users; Bhalla P et al.; Vaginal samples obtained from intrauterine contraceptive device (IUD) users with and without vaginitis and from healthy non-users were studied for presence of G . vaginalis and cytological changes . Vaginitis was present in 29.5 per cent IUD users and most of them (76%) yielded G . vaginalis . G . vaginalis was also recovered from asymptomatic IUD users and from non-users, but heavy colonisation was infrequent . The presence of clue cells and many small Gram negative bacilli and scanty lactobacilli were associated with G . vaginalis recovery . Inflammation was more frequent in IUD users than in non-users (P less than 0.001) . A significant correlation was also found between inflammatory changes and G . vaginalis colonisation among all groups of patients. Biomed Chromatogr, 1989 Mar, 3(2), 58 - 63 A rapid and specific HPLC-electrochemical method for the determination of endogenous 5-methyltetrahydrofolic acid in plasma using solid phase sample preparation with internal standardization; Lucock MD et al.; A rapid and specific HPLC-electrochemical method for determining endogenous 5-methyltetrahydrofolic acid (5MeTHF) in plasma is described . Quantitative solid phase extraction of 5MeTHF and internal standard, beta-hydroxyethyltheophylline, was carried out using proprietary phenyl bonded-silica columns (Bond Elut Phenyl cartridges, 1.0 mL capacity) . Chromatographic separation was achieved using a mobile phase consisting of 15% (v/v) methanol in 0.05 M KH2PO4, pH 3.5 at a flow rate of 2.0 mL/min in conjunction with a Waters Assoc . radially compressed Nova-Pak phenyl column (10 cm x 8 mm, 4 microns bonded silica) . The internal standard was measured by UV detection at 254 nm . A Bioanalytical Systems Inc . LC-17 glassy carbon oxidative flow cell with a potential held at +0.35 V vs Ag/AgCl using the LC-4A amperometric controller allowed levels of 1-2 ng/mL 5MeTHF to be measured in 500 microL of plasma . Daily appraisal of the ratio produced by authentic materials clearly demonstrated that quantitation using dual detection was not subject to problems of differential response . Inter-day variation of the differential detector response is cited . Comparison of the Lactobacillus casei bioassay with HPLC demonstrates good agreement between methods but at the same time highlights the drawback of using such non-specific methods to measure samples where more than one folylmonoglutamate may be present . Antoxidant free storage for three months at -70 degrees C in darkness resulted in no deterioration of 5MeTHF . A comparison of the means and range of values for plasma folate obtained using HPLC, L . casei bioassay and the radiometric binding assay is reported. J Clin Periodontol, 1989 Mar, 16(3), 175 - 8 Effect on plaque growth and salivary micro-organisms of amine fluoride-stannous fluoride and chlorhexidine-containing mouthrinses; Etemadzadeh H et al.; Amine fluoride-stannous fluoride mouthrinse was tested in a double-blind cross-over clinical trial on 16 healthy subjects . Chlorhexidine mouthrinse prepared in the same colour and taste was used as the control . After a preparatory phase of 2 weeks, during which the subjects' plaque and gingival indices were reduced to zero by professional cleaning, 1-week test periods without mechanical cleaning were started in random order . The fluoride test solution did not prevent plaque accumulation and growth of salivary S . mutans and lactobacilli as did chlorhexidine . Furthermore, the plaque wet weight was statistically highly significantly greater during test periods of fluoride rinsing than during the chlorhexidine phase . The cleansing effect, as subjectively estimated by questionnaire, was in favour of chlorhexidine, but discolorations of teeth were assessed as significantly worse during chlorhexidine periods . Neither of the test solutions showed any effect on salivary flow rate, pH, buffering capacity, or lysosomal enzyme activity. Mondo Ortod, 1989 Mar-Apr, 14(2), 201 - 5 {Oral hygiene motivation in orthodontic patients}; Sfondrini G et al.; The task of the present study has been to verify experimentally if prolonged motivation may produce better oral hygienic conditions in orthodontic patients . Applying oral hygienic index modified, gingival index and saliva lactobacilli's concentration count, the A.A . obtained encouraging data evaluating the motivated subjects situation versus control group. Aichi Gakuin Daigaku Shigakkai Shi, 1989 Mar, 27(1), 331 - 6 {Bacteriological examination of saliva of institutionalized severely mentally retarded persons with chronic rumination}; Ishiguro H et al.; The purpose of this study was to investigate dental caries prevalence, the pH and the presence of bacteria in the saliva of institutionalized severely mentally retarded persons with chronic rumination . The results obtained were as follows: The prevalence of dental caries among the group with chronic rumination was higher than among the group not suffering from rumination . The viable cell number of mutans streptococci, lactobacilli and yeasts in the group with chronic rumination was significantly higher than that of group without rumination . These results suggested that the saliva of the group with chronic rumination creates a favourable environment for aciduric bacterial growth. Cent Afr J Med, 1989 Mar, 35(3), 337 - 44 Microbial flora of the lower genital tract of women in labour at Harare Maternity Hospital . The Puerperal Sepsis Study Group; Mason PR et al.; The vaginal flora of 214 women who had been referred, in labour, to Harare Maternity Hospital was investigated by examination of vaginal washings and of cervical and urethral swabs taken before and/or after delivery . Four groups of patients were studied: women who had a normal vaginal delivery (NVD), women who were referred because of meconium stained liquor (MSL), women with a history of prolonged rupture of membranes (PROM), and women who were delivered by cesarean section (CS) . The first three groups had received no antibiotics during the 7 days preceding specimen collection, while specimens were collected from CS patients only after at least 48 hr of i.v . penicillin and chloramphenicol . T . vaginalis was identified in 19 percent of women, but was not associated with any specific patient group . Chlamydial antigen was detected in 13 percent of patients, but in only one patient (2 percent) in the MSL group . N . gonorrhoeae were isolated from 7 percent of women overall and 25 percent of the strains were penicillinase-producing . Gonococci were recovered significantly more frequently from the PROM patients than from NVD patients as were Group B streptococci and pigment-producing Bacteroides species . Lactobacilli were isolated from only 20 percent of women, despite the use of specific transport and isolation media for these organisms . Specimens from CS patients were taken after these had received parenteral penicillin and chloramphenicol and it was therefore not surprising to find major differences in their vaginal flora with a virtual absence of Gram-positive bacteria, and a high-rate of carriage of multi-resistant coliforms.(ABSTRACT TRUNCATED AT 250 WORDS) Kansenshogaku Zasshi, 1989 Mar, 63(3), 182 - 8 Primary resistance induced in mice by Lactobacillus casei following infection with herpes simplex virus; Watanabe T et al.; Primary host defense mechanism of mice against intraperitoneal infection with herpes simplex virus type 1 (HSV-1) was studied using heat-killed Lactobacillus casei YIT 9018 (LC 9018), a bacterial immunostimulant, in combination with inactivated HSV-1 antigen . Peritoneal exudate cells (PECs) in the early period of mice pretreated intraperitoneally (ip) with LC 9018 showed the cytotoxic activity against BALB/3T3 cells infected with HSV-1, in vitro, whereas PECs induced by thioglycollate broth did not . The limitation of HSV-1 replication in monolayers of HSV-1-infected PECs induced by LC 9018 was greater than that in PECs induced by thioglycollate broth . Both activities of PECs induced by LC 9018 were markedly enhanced by the administration of inactivated HSV-1 antigen which showed an interferon-producing activity in the early period after the treatment . These results suggest that the host defense mechanism of mice against intraperitoneal infection with HSV-1 may be mainly related to peritoneal macrophages activated by and interferon produced by the administration of LC 9018 and inactivate HSV-1 antigen. J Biol Chem, 1989 Feb 15, 264(5), 2625 - 33 Atypical transient state kinetics of recombinant human dihydrofolate reductase produced by hysteretic behavior . Comparison with dihydrofolate reductases from other sources; Appleman JR et al.; The transient state kinetics of catalysis for dihydrofolate reductase (DHFR) from several enzyme sources including highly purified recombinant human enzyme (rHDHFR) have been examined . Like DHFR from Escherichia coli, the enzyme from Lactobacillus casei, and isoenzyme 2 from Streptococcus faecium exhibit a slow increase in activity upon addition of substrates to enzyme . No slow hysteresis of this type was detected with recombinant human DHFR (rHDHFR) or DHFR from chicken or bovine liver or L1210 mouse leukemia cells (MDHFR) . In contrast, both rHDHFR and MDHFR exhibited a very rapid decrease in activity (t1/2 = 30 and 20 ms, respectively) during a phase that occurred after the first turnover of the enzyme but before establishment of the steady state . This intermediate phase was not observed for the bacterial enzymes or the avian enzyme, nor was it observed with a mutant of rHDHFR in which Phe-31 has been replaced by leucine . For rHDHFR the intermediate phase is not a consequence of product inhibition, substrate depletion, or enzyme instability . It may therefore be concluded that this unusual transient state kinetic behavior results from the existence of two conformers of the enzyme, one of which has a higher turnover number than the other with the equilibrium shifting in favor of the less active conformer during the course of catalysis . The equilibrium is particularly favorable for the less active conformer when NADP is present in the active site of rHDHFR, whereas bound tetrahydrofolate favors the more active conformer . The more active conformer has a 6-fold higher Km for dihydrofolate than does the less active conformer . The existence of these conformers is likely to produce cooperative behavior by rHDHFR in vivo. Biochemistry, 1989 Feb 7, 28(3), 1353 - 62 NMR studies of differences in the conformations and dynamics of ligand complexes formed with mutant dihydrofolate reductases; Birdsall B et al.; Two mutants of Lactobacillus casei dihydrofolate reductase, Trp 21----Leu and Asp 26----Glu, have been prepared by using site-directed mutagenesis methods, and their ligand binding and structural properties have been compared with those of the wild-type enzyme . 1H, 13C, and 31P NMR studies have been carried out to characterize the structural changes in the complexes of the mutant and wild-type enzymes . Replacement of the conserved Trp 21 by a Leu residue causes a decrease in activity of the enzyme and reduces the NADPH binding constant by a factor of 400 . The binding of substrates and substrate analogues is only slightly affected . 1H NMR studies of the Trp 21----Leu enzyme complexes have confirmed the original resonance assignments for Trp 21 . In complexes formed with methotrexate and the mutant enzyme, the results indicate some small changes in conformation occurring as much as 14 A away from the site of substitution . For the enzyme-NADPH complexes, the chemical shifts of nuclei in the bound coenzyme indicate that the nicotinamide ring binds differently in complexes with the mutant and the wild-type enzyme . There are complexes where the wild-type enzyme has been shown to exist in solution as a mixture of conformations, and studies on the corresponding complexes with the Trp 21----Leu mutant indicate that the delicately poised equilibria can be perturbed . For example, in the case of the ternary complex formed between enzyme, trimethoprim, and NADP+, two almost equally populated conformations (forms I and II) are seen with the wild-type enzyme but only form II (the one in which the nicotinamide ring of the coenzyme is extended away from the enzyme structure and into the solvent) is observed for the mutant enzyme complex . It appears that the Trp 21----Leu substitution has a major effect on the binding of the nicotinamide ring of the coenzyme . For the Asp 26----Glu enzyme there is a change in the bound conformation of the substrate folate . Further indications that some conformational adjustments are required to allow the carboxylate of Glu 26 to bind effectively to the N1 proton of inhibitors such as methotrexate and trimethoprim come from the observation of a change in the dynamics of the bound trimethoprim molecule as seen from the increased rate of the flipping of the 13C-labeled benzyl ring and the increased rate of the N1-H bond breaking. Br J Obstet Gynaecol, 1989 Feb, 96(2), 183 - 7 Vaginal pH in pregnancy in women delivered at and before term; Gleeson RP et al.; A microelectrode was used to measure vaginal pH in a longitudinal study during pregnancy in 30 women, 28 of whom were delivered at term . Excluding pH readings made in the presence of pathogenic organisms, 120 readings were taken . The mean vaginal pH of all subjects was 4.16 (SD 0.41) . Analysis of variance showed no significant within-subject variation of pH during pregnancy . Two women had spontaneous preterm deliveries (less than 37 weeks) and their mean vaginal pH during pregnancy was 5.05 and 5.06 . Eight women admitted in preterm labour, who then gave birth, had a mean pH of 5.43 (SD 0.94) which was significantly higher than the mean pH of 10 women in labour at term (4.58, SD 0.59) . The relation between bacterial growth in vitro and the surrounding pH was also examined . Escherichia coli, a potential pathogen, behaved differently from both Lactobacillus (a commensal organism) and Candida albicans . The clinical observations show a trend towards higher vaginal pH values in association with preterm labour. J Clin Microbiol, 1989 Feb, 27(2), 251 - 6 Prevalence of hydrogen peroxide-producing Lactobacillus species in normal women and women with bacterial vaginosis; Eschenbach DA et al.; A predominance of Lactobacillus species in the vaginal flora is considered normal . In women with bacterial vaginosis, the prevalence and concentrations of intravaginal Gardnerella vaginalis and anaerobes are increased, whereas the prevalence of intravaginal Lactobacillus species is decreased . Because some lactobacilli are known to produce hydrogen peroxide (H2O2), which can be toxic to organisms that produce little or no H2O2-scavenging enzymes (e.g., catalase), we postulated that an absence of H2O2-producing Lactobacillus species could allow an overgrowth of catalase-negative organisms, such as those found among women with bacterial vaginosis . In this study, H2O2-producing facultative Lactobacillus species were found in the vaginas of 27 (96%) of 28 normal women and 4 (6%) of 67 women with bacterial vaginosis (P less than 0.001) . Anaerobic Lactobacillus species (which do not produce hydrogen peroxide) were isolated from 24 (36%) of 67 women with bacterial vaginosis and 1 (4%) of 28 normal women (P less than 0.001) . The production of H2O2 by Lactobacillus species may represent a nonspecific antimicrobial defense mechanism of the normal vaginal ecosystem. J Bacteriol, 1989 Feb, 171(2), 940 - 6 Biosynthesis of linkage units for teichoic acids in gram-positive bacteria: distribution of related enzymes and their specificities for UDP-sugars and lipid-linked intermediates; Yokoyama K et al.; The distribution and substrate specificities of enzymes involved in the formation of linkage units which contain N-acetylglucosamine (GlcNAc) and N-acetylmannosamine (ManNAc) or glucose and join teichoic acid chains to peptidoglycan were studied among membrane systems obtained from the following two groups of gram-positive bacteria: group A, including Bacillus subtilis, Bacillus licheniformis, Bacillus pumilus, Staphylococcus aureus, and Lactobacillus plantarum; group B, Bacillus coagulans . All the membrane preparations tested catalyzed the synthesis of N-acetylglucosaminyl pyrophosphorylpolyprenol (GlcNAc-PP-polyprenol) . The enzymes transferring glycosyl residues to GlcNAc-PP-polyprenol were specific to either UDP-ManNAc (group A strains) or UDP-glucose (group B strains) . In the synthesis of the disaccharide-bound lipids, GlcNAc-PP-dolichol could substitute for GlcNAc-PP-undecaprenol . ManNAc-GlcNAc-PP-undecaprenol, ManNAc-GlcNAc-PP-dolichol, Glc-GlcNAc-PP-undecaprenol, Glc-GlcNAc-PP-dolichol, and GlcNAc-GlcNAc-PP-undecaprenol were more or less efficiently converted to glycerol phosphate-containing lipid intermediates and polymers in the membrane systems of B . subtilis W23 and B . coagulans AHU 1366 . However, GlcNAc-GlcNAc-PP-dolichol could not serve as an intermediate in either of these membrane systems . Further studies on the exchangeability of ManNAc-GlcNAc-PP-undecaprenol and Glc-GlcNAc-PP-undecaprenol revealed that in the membrane systems of S . aureus strains and other B . coagulans strains both disaccharide-inked lipids served almost equally as intermediates in the synthesis of polymers . In the membrane systems of other B . subtilis strains as well as B . licheniformis and B . pumilus strains, however, the replacement of ManNAc-GlcNAc-PP-undecaprenol by Glc-GlcNAc-PP-undecaprenol led to a great accumulation of (glycerol phosphate)-Glc-GlcNAc-PP-undecaprenol accompanied by a decrease in the formation of polymers. Zh Mikrobiol Epidemiol Immunobiol, 1989 Feb, (2), 3 - 8 {Mechanism of antagonistic activity of lactobacilli}; Tiurin MV et al.; Different species of lactobacilli, when grown together with salmonellae and shigellae, have been found to inhibit their growth and proliferation . The supernatants of the culture fluids of 8 species of lactobacilli have been shown to contain natural metabolites, other than lactic acid, with a wide spectrum of antibacterial activity with regard to different opportunistic aerobic and anaerobic microorganisms . The relationship between the antagonistic activity and the presence of plasmid DNA in a number of the most active antagonistic lactobacilli is discussed. Appl Environ Microbiol, 1989 Feb, 55(2), 461 - 4 Biotin-labeled plasmid DNA probes for detection of epithelium-associated strains of lactobacilli; Tannock GW; Biotin-labeled DNA probes prepared from whole plasmids (5.5 and 4.8 kilobases) of two lactobacillus strains (Lactobacillus delbrueckii 21 and Lactobacillus reuteri 100-23) were used to detect the homologous bacteria in microtome-cryostat-prepared sections of murine forestomach . The forestomach sections were incubated on nylon filter membranes placed on agar plates and, after lysis of the lactobacillus cells and denaturation of their DNA, were used in hybridization experiments with the strain-specific DNA probes . Hybridization of the probes to membranes containing sections from lactobacillus-free mice did not occur . The probes detected the presence of homologous strains of lactobacilli in sections cut from the forestomach of mice harboring one or both of the strains. Appl Environ Microbiol, 1989 Feb, 55(2), 279 - 83 Colonization of the porcine gastrointestinal tract by lactobacilli; Pedersen K et al.; Eight strains of lactobacillus isolated from the porcine gastrointestinal tract were tested for their ability to adhere in vitro to cells collected from stratified squamous epithelium in the digestive tracts of newborn piglets . Piglets were inoculated with individual strains, and their digestive tracts were sampled at intervals to determine the colonizing ability of the lactobacilli . The results of the in vitro test did not predict whether a lactobacillus strain would associate with stratified squamous epithelium in the piglet digestive tract, but epithelial association in vivo appeared to be an important factor in the maintenance of lactobacillus populations in the tract . None of the lactobacillus strains used as inocula was numerically dominant in the tract 7 days after inoculation of the piglets with a single dose of the bacteria. Scand J Dent Res, 1989 Feb, 97(1), 25 - 32 Systematic analysis of gingival state and salivary variables as predictors of caries from 5 to 7 years of age; Sullivan A et al.; The aim was to assess the caries predictive ability of gingival state, salivary counts of lactobacilli and S . mutans, salivary secretion rate and buffer capacity of the saliva from the ages of 5 to 7 yr . The material comprised 105 children who were examined once a year . All available screening levels were systematically tested for the single variables as well as for combinations of two or three variables . The screening levels were set where the highest possible sensitivity and specificity were reached simultaneously . The predictive values for positive and negative tests were then calculated . The S . mutans count turned out to have the best combination of sensitivity (0.41) and specificity (0.83) . The single variables gingival state and S . mutans had constant screening levels, while the values for lactobacilli and buffer capacity varied . The saliva secretion rate could not be measured at 5 yr of age . Systematic analyses of predictive ability of combinations of two or three variables were then performed and did not result in any clinically significant improvements of the predictive ability . The observed low predictive ability might be explained by the fact that 1) many children changed their habits during the period of investigation and 2) they were examined and treated once a year according to their individual needs, which meant variations in conservative as well as in preventive treatment. Infect Immun, 1989 Feb, 57(2), 559 - 65 Control of Escherichia coli populations by a combination of indigenous clostridia and lactobacilli in gnotobiotic mice and continuous-flow cultures; Itoh K et al.; The function of indigenous lactobacilli in the control of other intestinal microbial species is not clear . Still more controversial is the effect of dietary bacterial supplements containing lactobacilli or other species . This situation is unlikely to change unless the mechanisms that control the colonization of ingested bacteria are better understood, and until more detailed information becomes available on the mechanisms by which certain populations of indigenous bacteria can affect the population sizes of other species . We used gnotobiotic mice and a continuous-flow culture system to study the interactions between Escherichia coli and (i) clostridia (in chloroform-treated cecal suspensions from conventional mice) and (ii) three strains of lactobacilli isolated from conventional mice . In gnotobiotic mice, the lactobacilli suppressed E . coli multiplication in the stomach and the small intestine, but had no demonstrable effect on E . coli multiplication in the large intestine . In contrast, clostridia were most effective in controlling E . coli multiplication in the large intestine . In the presence of both lactobacilli and clostridia, E . coli populations in the various regions of the gastrointestinal tract resembled those found in conventionalized control animals . The control of E . coli populations was not related to changes in pH or intestinal motility . In vitro stimulation of the above-described in vivo interactions required a two-stage continuous-flow culture in which the effluent from the first stage represented the influx to the second . The first stage was inoculated with lactobacilli, and the second stage was inoculated with either a pure culture of E . coli or E . coli and clostridia . In these instances, the E . coli populations in the second stage of the culture resembled in size those found in the large intestine of gnotobiotic mice harboring a similar flora . Although there are some current shortcomings of this in vitro model, we expect that a multistage continuous-flow culture can be developed to satisfactorily model the interactions among bacterial populations along the entire gastrointestinal tract. Indian J Exp Biol, 1989 Feb, 27(2), 141 - 5 A cytotoxic substance produced by a wild culture of Lactobacillus casei D-34 against tumour cells; Manjunath N et al.; A wild lactic culture isolated from dahi (fermented milk) sample and characterised as L . casei D-34 was found to be significantly cytotoxic (34-36%) against three tumour cell lines, HeLa, HEp-2 and HFS-9 . The cytotoxic substance (CS) was found to be in the culture supernatants, protein in nature, with a molecular weight ranging from 17,000-20,000 . The crude culture supernatant was partially purified by dialysis and ion exchange chromatography as anionic, cationic and neutral fractions . Among the fractions, except for the anionic fraction, others were found to be highly cytotoxic against all three tumour cell lines . The cationic, neutral and pooled (anionic:cationic:neutral in 1:1:1 ratio) fractions showed 50, 70, 70% cytotoxicity against Hep-2 cells, 70, 88, 94% against HFS-9 cells and 50, 89, 90% against HeLa cells respectively . Pooled fraction was found to exhibit higher percent of cytotoxicity compared to individual fractions indicating a synergistic effect . (3H)-thymidine incorporation studies revealed that CS and its fractions inhibited DNA synthesis in tumour cells . The CS was stable towards heat and pH changes. Zentralbl Bakteriol Mikrobiol Hyg {A}, 1989 Jan, 270(3), 385 - 95 {Microbial colonization of carious progression stages in the dentin of human teeth--a controlled therapeutic study}; Kneist S et al.; In a controlled clinical trial the microflora of the cavity floor of 70 primary lower second molars with deep carious lesions were determined after caries excavation . The teeth were extracted and pulpal status was evaluated after 16 months of microbial control to determine the etiopathogenic role of germs for carious progression in dentine . 67% of the primary molars were free from pulpal inflammations . Soft carious dentine were significantly higher infected than the clinically acceptable hard dentine . Only in 40% of the cavity floors the microorganisms were eliminated . In the infected teeth basophilic microorganisms were found in causality to pulps without inflammations; acidogenic streptococci and lactobacilli were involved in pulpal inflammations . Results indicate that the latter genera of microorganisms are of etiological significance for carious progression in dentine. Caries Res, 1989, 23(1), 49 - 54 Caries-related microbiological findings in a group of teenagers and their parents; Alaluusua S et al.; The levels of salivary Streptococcus mutans and lactobacilli and caries experience were studied in 113 teenagers and 163 adults . The study population consisted of 82 mother-child and 73 father-child pairs . The number of decayed, missing, and filled tooth surfaces (DMFS) was eightfold higher in adults than in teenagers (56.4 +/- 22.8 vs . 7.3 +/- 6.7) . The percentage distribution of the level of salivary S . mutans and lactobacilli was approximately the same in both groups . The mean number of DMFS increased with increasing levels of salivary S . mutans and lactobacilli, the correlation being highly significant both in teenagers and adults . There was a significant correlation of the DMFS indices in the mother-child pairs (r = 0.364), but the correlation was not significant in the father-child pairs (r = 0.138) . The salivary level of S . mutans was higher in the children of mothers with high DMFS values compared to the children of mothers with low DMFS values. Biull Eksp Biol Med, 1989 Jan, 107(1), 36 - 9 {Primary structure of histidine decarboxylase}; Prozorovskii VN et al.; The results of investigation of the primary structure of the Histidine Decarboxylase Micrococcus sp . n . are reported . A comparison of the primary structure of the Histidine Decarboxylase Micrococcus sp . n . with that of the Lactobacillus 30a enzyme suggests the alignment with a 52% identity . It is therefore highly probable that two proteins have evolved from common ancestry . The conservative amino acid sequences with residues (pyruvate, cysteine) of the active center have been found. Swed Dent J, 1989, 13(4), 125 - 30 Results with the caries activity test "Cariostat" compared to prevalence of mutans streptococci and lactobacilli; Camling E et al.; The colorimetric test, Cariostat (Sankin, Japan), for measuring the caries activity, based on the presence of acidogenic microorganisms, was evaluated . The results were compared with the prevalence of mutans streptococci and lactobacilli in the samples . Pure cultures of S . mutans and lactobacilli were inoculated into the test medium . A maximum color change from blue to yellow was not obtained not even when large numbers of bacteria were used . Samples of stimulated saliva and plaque samples from people with different salivary levels of mutans streptococci and lactobacilli were inoculated into the test medium . Samples with very low or very high numbers of cariogenic bacteria were found in all score groups indicating that the Cariostat test did not satisfactorily differentiate samples with low or high numbers of microorganisms associated with dental caries. Rev Belge Med Dent, 1989, 44(1), 37 - 45 {The lacto-tampon concept . Determination and prevention of an increased risk for caries}; Wikner S; The lactobacillus count and buffer capacity (final pH) of whole stimulated saliva was assessed and related to the caries increment over the next 12 months in 327 teenagers . The caries increment was 3-4.5 times higher in children having a high lactobacillus count combined with a low buffer capacity, compared with other lacto-buffer combinations (P less than, 01-,001) . The protective influence of a high buffer capacity seems to be stronger than the caries-promoting influence of high sugar consumption, indicated by a high lactobacillus count . On the one hand a low sugar consumption seems to have a decisive determining preventive effect even in children having impaired buffer systems . On the other hand a high sugar consumption strongly promotes caries when the buffer capacity is low . It is also demonstrated that consumption changes during a longitudinal study may impair the relationships between the salivary parameters, assessed at baseline, and the caries increment during the study. Cancer Chemother Pharmacol, 1989, 23(6), 353 - 7 Accumulation of tetrahydrofolates in human plasma after leucovorin administration; Bunni MA et al.; Reduced folates in plasma after i.v . and oral leucovorin administration were estimated by a ternary complex assay based on the incorporation of CH2FH4 into a stable complex with Lactobacillus casei thymidylate synthase and {3H}FdUMP . Each of the reduced folates, CH2FH4, FH4, and 5CH3FH4, could be quantitatively recovered from plasma by this approach even in the presence of high concentrations of the parent compound leucovorin . Examination of the accumulation kinetics of these reduced folates showed that after i.v . administration of 20 mg D,L-leucovorin to a healthy volunteer, FH4 and, to a lesser extent, CH2FH4 accumulated to maximal levels very early (less than 15 min), with a subsequent depletion that had a half-life of approximately 30 min . Accumulation of FH4 reached a peak level that was 12% of the maximal level of 5CH3FH4 achieved and more than 3 times greater than the pretreatment level of this common, circulating reduced folate form . Similar accumulation patterns were observed in a female patient with metastatic colonic cancer who was undergoing methotrexate (MTX)/fluorouracil therapy followed by i.v . leucovorin (15 mg) . FH4 also accumulated, but to a lesser extent and over a longer period of time, when the same dose of leucovorin given orally . When several similar doses of leucovorin were given prior to the experimental dose, greater accumulation and duration of the FH4 response was observed . We propose that accumulation of FH4 and CH2FH4 could provide a circulating source of the reduced folate thought to be the active form for both high-dose MTX with leucovorin rescue and enhancement of fluorouracil activity. Swed Dent J Suppl, 1989, 57, 1 - 40 On dental health in Icelandic children . Observations during a clinical dentifrice trial; Bjarnason S; Dental health and the relative caries preventive effect of different fluoride dentifrice formulations were assessed in an Icelandic child population . For a cross sectional study 1370 children, aged 11 and 12 years were examined for caries and periodontal conditions employing standardized methods . Longitudinal data were obtained from 1161 children, participating in a three-year dentifrice trial . Randomly selected subgroups were employed to study salivary levels of mutans streptococci and lactobacilli, consumption of high-sucrose products and enamel fluoride content in relation to caries experience . Relative caries preventive effect of anticalculus agents (HEBP and AHBP) and lowered fluoride concentration (250 ppm) in dentifrices were evaluated in a three-year double blind randomized clinical trial . At the baseline examination a comparatively high mean caries prevalence (DFS 9.9) was registered . Longitudinal observations showed a mean caries increment of 10.5 surfaces during a three-year period . Simultaneously an improvement of gingival health was observed . A majority of children harbored mutans streptococci (98%) and lactobacilli (92%) . High numbers of either microorganism were associated with high caries prevalence . S . sobrinus carriers were more frequent among children with high numbers of CFU per ml saliva . The consumption of sugar-containing products was generally frequent . Higher caries increment was associated with consumption of sweets and bakery products during mealtimes . The fluoride content of surface enamel was comparatively low, reflecting scarce post eruptive exposure . Dentifrice, containing 250 ppm fluoride showed significantly lower caries preventive effect compared to 1000 ppm sodium fluoride and MFP formulations . Addition of anticalculus agents (biphosphonates) had no negative effect on reducing caries increment . These studies show that dental caries constitutes a major health problem among Icelandic children, where high sugar consumption and comparatively low exposure to fluoride appear to play major roles . The use of fluoride dentifrice has not been able to limit caries development. Swed Dent J, 1989, 13(3), 103 - 9 Dental health status in Latin-American preschool children in Malmö; Vidal OP et al.; Seventy-three 3-6-year-old preschool children from Latin-American countries living in Malmo were examined in order to evaluate their oral health status . The children had on average 3.5 dmf teeth and 4.9 dmf surfaces . Forty-one per cent were caries-free . Oral hygiene was satisfactory in 25% of the children . Regarding dietary habits, 41% of the children had more than two unsuitable intakes per day . Night-time use of a nursing bottle with sugar-containing beverage was reported for 30% . The mean dmfs values differed significantly between children with and without S . mutans and lactobacilli in their saliva . Oral health status deviated markedly from the total group of 3-6-year-olds in Malmo, which might be explained by differences in cultural background and the ability to speak and understand Swedish . In conclusion, the Latin-American preschool children and their families should be regarded as a risk group for caries and gingivitis and consequently be subjected to special preventive dental care. Microbiol Immunol, 1989, 33(1), 23 - 34 Comparative characterization of spirosomes isolated from Lactobacillus brevis, Lactobacillus fermentum, and Lactobacillus buchneri; Nomura S et al.; Spirosomes, cytoplasmic fine spirals, were isolated and purified from Lactobacillus brevis ATCC 8287, L . fermentum F-1, and L . buchneri ATCC 4005, and their morphological, biochemical, and immunological properties were investigated . The spirosomes of these lactobacilli were morphologically indistinguishable from one another, and they had the same buoyant density of 1.320 g/cm3 in CsCl . All of the spirosomes were composed of a single protein, spirosin, with an apparent molecular weight of about 95,000 for L . brevis and L . fermentum and of about 96,000 for L . buchneri as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) . The spirosins from the three lactobacilli were compared by peptide mapping on SDS-PAGE after cleavage with N-chlorosuccinimide and limited proteolysis with Staphylococcus aureus V8 protease . The peptide map of the L . brevis spirosin was identical with that of the L . fermentum spirosin, whereas it was markedly different from the L . buchneri spirosin . The amino acid composition of the L . brevis spirosin was almost similar to that of the L . fermentum spirosin, while it differed appreciably from the L . buchneri spirosin . Using antiserum against the L . brevis spirosin, immunodiffusion test revealed that the antigenicity of the spirosomes from L . brevis was identical with that from L . fermentum, whereas it was partially different from that from L . buchneri. Plasmid, 1989 Jan, 21(1), 9 - 20 Characterization of a gram-positive broad-host-range plasmid isolated from Lactobacillus hilgardii; Josson K et al.; Two plasmids, pLAB1000 and pLAB2000 (3.3 and 9.1 kb, respectively), have been isolated from a grass silage strain of Lactobacillus hilgardii . Both plasmids were cloned in Escherichia coli and characterized through restriction mapping . A 1.6-kb XbaI-SacI fragment of pLAB1000 appeared to be sufficient for autonomous replication in Lactobacillus plantarum and in Bacillus subtilis . Different shuttle vectors for E . coli and gram-positive bacteria were developed using the pLAB1000 plasmid . These could stably be maintained in Lactobacillus, Enterococcus, and Bacillus under selective conditions . Plasmids sharing DNA homologies with pLAB1000 have been observed in different strains of the related species L . plantarum. Arzneimittelforschung, 1989 Jan, 39(1), 12 - 5 Alpha-monoamides of methotrexate as potential prodrugs; Antonjuk DJ et al.; alpha-Monoamides of methotrexate were evaluated for their potential as prodrugs . Studies on 11 alpha-monoamides and 5 gamma-monoamides of methotrexate showed that the gamma-monoamides were about as strong inhibitors of Lactobacillus casei dihydrofolate reductase as methotrexate, while I50 of the alpha-monoamides were 1-2 orders higher . The concentration for growth inhibition of murine L1210 cells for methotrexate gamma-propylamide and alpha-propylamide were respectively 1-2 and 2-3 orders higher than that of methotrexate . In contrast, only alpha-monoamides caused significant increase in life span of mice with transplanted L1210 leukaemia, the highest effect being given by the alpha-propyl and the alpha-butylamide . The possibility that the in vivo activity of the alpha-monoamides might be related to in vivo transformation to methotrexate was studied by HPLC analysis of mice serum after administration of the alpha- and gamma-propylamides. Microbiologica, 1989 Jan, 12(1), 69 - 74 Antagonism and adhesion among isogenic strains of Lactobacillus reuteri in the caecum of gnotobiotic mice; Sarra PG et al.; Interactions between a wild adhesive Lactobacillus reuteri strain isolated from chicken crop and its isogenic plasmid-free derivative were studied . Antagonism between these strains was tested in vitro or associated with gnotobiotic mice . The cured strains repressed the establishment of the wild, plasmid bearing strain in the gastrointestinal tract of gnotobiotic mice . Interactions between isogenic strains were independent of the amount of cells administered to mice . The cured derivative strain maintains its specific ability to adhere to epithelial cells of chicken crop. Med Dosw Mikrobiol, 1989, 41(3-4), 206 - 14 {Diagnostic and clinical characteristics of mycotic vaginitis caused by Candida glabrata (Torulopsis glabrata)}; Pawlik B et al.; In this study an evaluation of frequency of occurrence of C . glabrata, its diagnosis, sensitivity to antifungal drugs and its significance in pathogenesis of mycotic vaginosis was performed . Strains belonging to C . glabrata genus constituted 12.1% of total of 852 isolated strains and 39.2% of strains other than C . albicans . During fungal vaginosis caused by C . glabrata Lactobacillus sp . was present and normal pH values of vaginal secretion were seen . In direct preparations single or few leukocytes were observed and usually numerous blastospores were present . During evaluation of the sensitivity of C . glabrata strains to antimycotic agents a decreased sensitivity of these strains to clotrimazole and ketoconazole was found what speaks for their low usefulness in the treatment of mycotic vaginosis . Significance of C . glabrata in pathogenesis of mycotic vaginosis is not questioned since release of complaints and clinical symptoms in patients with positive therapeutic effect is seen and their persistence in a group of patients with treatment failure. Mol Biother, 1989, 1(5), 279 - 82 Antitumor effect of lactobacilli substances: "L . bulgaricus effect"; Clemmesen J; Scattered reports suggest a moderate effect, difficult to reproduce, of products from strains of specific lactobacilli (L . bulgaricus) used for yogurt on cancer and leukemia in mice . Skeptics have disregarded that even lyophilized preparations of demonstrated activity will lose effect when stored above -80 degrees C . This explains some inconsistencies of results and difficulties in repetition . Flow DNA fluorometry of ascitic fluid from leukemic mice has shown some accumulation of cells in the G2 + N phase after 8 hours, clearly different from the effect of 5-fluorouracil, although occasionally similar in strength . The effect depends on the strain of lactobacilli, and seems to deserve rational analysis by modern methods. Pediatrician, 1989, 16(3-4), 156 - 60 Specific microorganisms and dental caries in children; Krasse B; During the last decades it has been convincingly demonstrated that mutants streptococci and lactobacilli are of etiologic importance in dental caries . The knowledge has been applied in several clinical studies and the results show that microbiological examinations can be valuable adjuncts in the diagnosis, treatment and prevention of dental caries . Although dental caries has declined considerably, some children still develop high caries activity . It is important that such children are identified and given preventive treatment before they develop rampant decay . In this context pediatricians play an important role and the new microbiological knowledge can be a valuable tool. Arch Oral Biol, 1989, 34(1), 43 - 53 Arginolytic and ureolytic activities of pure cultures of human oral bacteria and their effects on the pH response of salivary sediment and dental plaque in vitro; Wijeyeweera RL et al.; Thirty-nine different microorganisms commonly found in supragingival plaque and salivary sediment were screened for their ability to raise the pH by producing base from arginine, lysylarginine and urea . Only Actinomyces naeslundii and Staphylococcus epidermidis showed significant pH-rise activity with all three compounds . Eleven bacteria demonstrated such activity with arginine and lysylarginine but not with urea . Only one, Actinomyces viscosus, produced a pH-rise with urea but not with the two arginine compounds . The remaining 26 bacteria showed little or no base-forming activity with any of the three test substrates . The ability of the different oral bacteria to produce base (especially from urea) was a less universal function than their ability to produce acid from fermentable carbohydrate . Substituting pure cultures of arginolytic or non-arginolytic bacteria for portions of the mixed bacterial populations of plaque or sediment in test incubations containing glucose and arginine altered their ability to produce pH-fall-pH-rise responses shaped like those of the Stephen curve in vivo . In general, addition of arginolytic bacteria made these in vitro pH responses less acidic, whereas addition of non-arginolytic bacteria made the responses more acidic . Because of the relatively high arginolytic activity of the plaque harvested in this study, the effect of adding non-arginolytic bacteria was more readily seen than the converse . Similar changes in levels of ureolytic microorganisms and incubation with glucose and urea had little effect on sediment or plaque being able to produce a pH-fall-pH-rise type of response . When increasing proportions of the mixed bacteria in salivary sediment were replaced with the highly cariogenic Lactobacillus casei or Streptococcus mutans, the pH minimum became slightly more acidic and then slightly more alkaline, whereas the pH-rise became progressively and significantly less . Thus arginolytic bacteria have a different and greater effect on shaping the pH response of plaque or sediment than ureolytic bacteria . A large change in the proportions of arginolytic or non-arginolytic microorganisms may be needed to make a plaque microflora potentially non-cariogenic or cariogenic, respectively. Med Dosw Mikrobiol, 1989, 41(1), 60 - 6 {Occurrence and the sensitivity to drugs of fungi other than Candida albicans isolated from the vagina}; Pawlik B et al.; Frequency of occurrence of fungal species distinct from C . albicans isolated from vaginal mucosa and their sensitivity to antimycotic chemotherapeutics was determined . Material consisted of 452 fungal strains isolated from vagina from patients suffering from afflictions within genital area . Fungal strains isolated belonged to 13 genera . Fungi distinct from C . albicans constituted 27.1% of all the strains . Fungi the most frequently isolated from vagina belonged to following genera: T . glabrata 35.2% C . krusei 18.4% C . pseudotropicalis 15.2% S . cerevisiae 10.4% . In the majority of cases of vaginal infections caused by fungi distinct from C . albicans, Lactobacillus sp . was present and normal pH values of vaginal content 3/4 with variable number of leucocytes were observed . Evaluation of sensitivity to antimycotic drugs of fungal strains was performed by agar dilution technique . In this study the following chemotherapeutics were assayed: nystatin, pimaricin, amphotericin B, flucytosine, clotrimazole, miconazole and ketoconazole . It is worth to underline resistance of T . glabrata and S . cerevisiae to clotrimazole and ketoconazole . Moreover, resistance of strains belonging to genera C . krusei and C . pseudotropicalis to amphotericin B and C . krusei strains to nystatin and flucytosine was noted. Microbiol Immunol, 1989, 33(10), 797 - 810 Endotoxic properties of chemically synthesized lipid A analogs . Studies on six inflammatory reactions in vivo, and one reaction in vitro; Yoshida M et al.; Biological activities of two groups of synthesized lipid A analogs, the counterpart of biosynthetic precursor, Lehmann's Ia type, 406, and E . coli lipid A type, 506, as well as their non-phosphorylated, and mono-phosphorylated analogs were investigated . The activities employed included four bone marrow cell reactions in mice, mice skin reaction, leukocytes migration in rabbits' cornea, and hemagglutination . Compound 406 and 506 elicited bone marrow reactions in mice and hemagglutination of mouse RBC, although 406 failed to elicit hemorrhage and necrosis also in mice skin . Compound 406 did not elicit corneal reaction in rabbits . The results suggest that for elicitation of this reaction and mice skin reaction, acyloxyacyl structure is required . Cytotoxicity and thromboplastin production of four bone marrow reactions had been reported by us to be endotoxic reactions, since these had not been elicited by peptidoglycan of Lactobacillus and Staphylococcus (1981) and 300 series synthesized analogs (1984) which did not have endotoxic structures . From these results, it seems that these two marrow reactions and hemagglutination require, as does the limulus test, the lipid A part structure as is present in 406. Indian J Exp Biol, 1989 Jan, 27(1), 72 - 5 Cytotoxic activity of lactobacilli isolated from plant and dairy sources; Manjunath N et al.; Forty four cultures of Lactobacilli isolated from their natural sources such as dahi, raw milk and fermenting rice-pulse doughs etc . along with four standard strains of Lactobacilli were assayed for their cytotoxic activity against three secondary tumour cell lines . Three cultures isolated from dahi samples and identified as Lactobacillus casei D-34, L . casei D-48a, L . plantarum D-70a along with one standard strain L . casei B 1922 exhibited significant cytotoxic activity in the range of 30 to 36%. Biotherapy, 1989, 1(3), 169 - 77 Augmentation of resistance of mice to bacterial infection by a polysaccharide-peptidoglycan complex (PSPG) extracted from Lactobacillus casei; Nomoto K et al.; A water-soluble polysaccharide-peptidoglycan complex (PSPG) was prepared from heat-killed Lactobacillus casei by digesting the bacteria with N-acetylmuramidase . The molecular weight of PSPG was over 30,000, and the polysaccharide portion of PSPG, its main component was composed of rhamnose, glucose, galactose, glucosamine and galactosamine . Mice pretreated intraperitoneally with PSPG survived after a lethal infection with Listeria monocytogenes or Pseudomonas aeruginosa . The growth of infecting bacteria (L . monocytogenes, P . aeruginosa, Salmonella typhimurium, Escherichia coli) in both the peritoneal cavity and the liver was inhibited markedly in the mice that had been treated with PSPG . It was suggested that macrophages may be the main effector for the anti-infectious effect of PSPG since treatment of mice with carrageenan, a selective macrophage blocker, markedly reduced the anti-infectious effect of PSPG. Adv Enzyme Regul, 1989, 29, 189 - 207 A rapid, reliable, and convenient method for the purification of thymidylate synthase from amethopterin-resistant Lactobacillus casei; Zapf JW et al.; The results presented herein describe the development of a new method for the purification of thymidylate synthase from amethopterin-resistant L . casei . This method includes three chromatographic steps: batchwise CM-Sephadex, Q-Sepharose, and 10-formylfolate-Sepharose affinity steps, and the whole procedure can be performed in three days . Additionally, the procedure has consistently produced enzyme with a specific activity of 3.0 to 3.2 . The first step, the batchwise purification of L . casei cell free extracts on CM-Sephadex, resulted in a 120-fold purification with 115% recovery of the activity applied to the resin . The Q-Sepharose purification step yielded a 1.5-fold purification producing enzyme with a specific activity of 2.4 units/mg . As the final step in the sequence, 10-formylfolate-Sepharose affinity chromatography yielded enzyme with a specific activity of 3.0-3.2 units/mg . The entire procedure provided a 230-fold purification with 67% recovery of the activity originally present in the cell free extract . However, the purified enzyme required extensive dialysis to achieve maximal activity . The data presented also clearly demonstrate the resolution of thymidylate synthase on the 10-formylfolate affinity column into forms which have different specific activity values and FdUMP binding ratios . Generally, enzyme with low activity leaches off this affinity matrix first, while enzyme with high activity elutes off last . These results are in accord with the sensitivity of the dimeric thymidylate synthase active site cysteines to oxidization . Consequently, when one of the two active site cysteines is oxidized, the enzyme loses the ability to bind FdUMP at this active site and its specific activity is proportionally lower . Thus, pure thymidylate synthase protein exists as a mixture of heterogeneous activity states . This condition is best examined by performing native PAGE on enzyme that has been incubated with excesses of FdUMP and CH2H4folate. Adv Enzyme Regul, 1989, 29, 141 - 57 Methotrexate 5-aminoallyl-2'-deoxyuridine 5'-monophosphate: a potential bifunctional inhibitor of thymidylate synthase; Stuhmiller LM et al.; Mercuration of 2'-deoxyuridine 5'-phosphate (dUMP) followed by alkylation with allylamine in the presence of K2PdCl4 afforded the 5-aminoallyl deoxynucleotide, which was isolated by sequential Dowex 50 H+ and DEAE-Sephadex chromatography . Further reaction of the product with the N-hydroxysuccinimide ester of methotrexate (MTX) in dry dimethyl sulfoxide gave an MTX-aminoallyl-dUMP covalent complex separable by DEAE-Sephadex chromatography . Reprecipitation with acid from basic solution offered further purification and the structure was confirmed by elemental analysis, NMR and absorbance spectra . The product was an inhibitor of rat liver dihydrofolate reductase (I50 approximately 250 nM, cf . MTX I50 approximately 60 nM) and Lactobacillus casei thymidylate synthase . With the latter enzyme, inhibition was competitive with both nucleotide and folate substrates (Ki = 2.6 and 3.5 microM, respectively) and partial enzyme-inhibitor binary complex could be detected by gel electrophoresis . Large fluorescence changes were observed on titration of the synthase with MTX-aminoallyl-dUMP and alterations in the UV difference spectra similar to those seen on titration of the enzyme with MTX were also noted . The compound was a poor growth inhibitor for cultured murine L1210 and human CCRF-CEM cell lines, which probably reflects low cellular uptake. Adv Enzyme Regul, 1989, 28, 3 - 11 Visualization of membrane-associated folate transport proteins; Pope LE et al.; Transport of Methotrexate (MTX) into cells, via the "reduced folate" transport system, is a critical factor in the effectiveness of the drug in cancer chemotherapy, and defective transport is one of the principal types of resistance to MTX . Probes capable of detecting membrane-associated folate transport proteins (ftp's) in individual cells are potentially useful for identifying structural and functional domains and for investigating mechanisms of substrate translocation . Polyclonal antibody to highly purified ftp from Lactobacillus casei, in conjunction with a second, gold-labeled antibody, has been used to visualize, via electron microscopy, the protein in Triton-treated membrane fragments and in the membrane and cytoplasm of spheroplasts . To visualize ftp in L1210 cells, the substrate-binding site was first labeled covalently with activated fluorescein-Methotrexate, and the cells were then treated with anti-fluorescein antibody and the gold-labeled antibody. Int J Biochem, 1989, 21(11), 1217 - 21 Inhibition of thymidylate synthase by pyridoxal phosphate; Chen SC et al.; 1 . Pyridoxal phosphate (PLP) reversibly inhibited thymidylate synthase from Lactobacillus casei with a KI of 0.6-0.9 microM . 2 . The inhibition was competitive with dUMP and noncompetitive with 5,10-methylenetetrahydrofolate which is consistent with an ordered addition of substrates . 3 . The spectrum of PLP was altered by the addition of thymidylate synthase . The spectral changes suggest formation of a thiohemiacetal with an enzyme sulfhydryl group rather than Schiff base formation with a lysine side chain. Int J Vitam Nutr Res, 1989, 59(2), 147 - 50 Comparison of radiometric and microbiological assays for the determination of folate in fortified gruel and porridge; Osterdahl BG et al.; The results obtained by a radiometric assay of folate in fortified gruel and porridge, using a commercially available radioassay kit, were compared with those obtained by a standard microbiological assay with Lactobacillus casei . A lower level of folate was found with the radiometric assay . Comparison of the results obtained with the two methods gave a correlation coefficient of 0.991 when the analysis was performed on samples extracted at the same time . A lower correlation coefficient was obtained between the result if the extractions were carried out on different occasions . The radiometric assay is a acceptable method for the routine determination of folate in fortified gruel and porridge. Caries Res, 1989, 23(5), 351 - 7 Cariological assessment of leucrose {D-glucopyranosyl-alpha(1----5)-D-fructopyranose} as a sugar substitute; Ziesenitz SC et al.; Leucrose {D-glucosyl-alpha(1----5)-D-fructopyranose} prepared by microbial-enzymatic transglycosidation from sucrose, is the first alpha(1----5)-linked disaccharide which possesses excellent nutritional properties with regard to metabolic utilization and is well tolerated . The aim of the present work was to assess its cariogenic potential . Yeast invertase was shown to be inhibited by leucrose in a noncompetitive way, while hydrogenated leucrose (leucritol) acted as an activator . Plaque polysaccharide forming glucosyltransferases from Streptococcus cricetus AHT were not influenced by leucrose, but by leucritol . Essentially no acid formation was observed after incubation of leucrose with suspensions of human dental plaque, S . mutans NCTC 10449, Lactobacillus casei LSB 132 and Actinomyces viscosus Ny 1 No . 30 . Leucrose was a competitive inhibitor of the acid formation from sucrose by S . mutans NCTC 10449 at neutral pH . Furthermore, leucrose inhibited at neutral pH considerably the uptake of sucrose by S . mutans NCTC 10449 . The uptake of fructose and maltose was also inhibited but that of glucose not at all . In Cara rats as the animal model, leucrose was compared to sucrose and to corn starch for its cariogenic potential . In sharp contrast to the group fed with 30% sucrose, caries scores of the 30% leucrose group were not significantly different from the starch group . pH telemetry with an indwelling electrode in man proved lueucrose to be 'safe for teeth' since plaque pH did not drop below pH 5.7 . Leucrose is a novel noncariogenic disaccharide and thus represents a highly promising sugar substitute for caries prevention. Pediatriia, 1989, (6), 24 - 9 {Effects of dysbacteriosis and impairment of immunity formation in the early neonatal period on morbidity of children during the 1st year of life and ways of its reduction}; Chernyshova LI; The catamnesis of 124 infants has been studied in the course of 12 months by means of questionnaires . Dysbacteriosis that occur in the early neonatal period produce an unfavourable effect on the immunity establishment and increase the disease incidence among the neonates . Moreover, dysbacteriosis may later result in the impairment of the formation of tolerance to food antigens supplied to the gastrointestinal tract . The carriership of Staphylococcus aureus in the fauces occurring in the presence of the decreased characteristics of local defence and phagocytic system of the newborn favours an increase of the disease incidence in the babies as well as the formation of the chronic foci of infection in the future . Prevention of dysbacteriosis of the early neonatal period by means of the use of biopreparations, particularly of pure cultures of Lactobacillus acidophilus does not only reduce the disease incidence in children of the first year of life but also improves on the whole their health status. Pediatriia, 1989, (3), 25 - 30 {Prevention of dysbacteriosis in the early neonatal period using a pure culture of acidophilic bacteria}; Moshchich PS et al.; In order to prevent dysbacterioses occurring in the early neonatal period, 60 neonates classified with the risk group were administered, when they were still at the puerperal room, pure cultures of specially selected strains of Lactobacillus acidophilus possessing marked anti-Klebsiella activity and antagonistic activity against Staphylococcus aureus . The cultures were instilled into the mouth and nasal passages . Forty neonates administered bifidum-bacterin and 54 neonates not given biopreparations were used as control . Application of the pure culture of Lactobacillus acidophilus noticeably raised the efficacy of the preventive measures . Over 60 percent of the neonates were discharged from the maternity home with a normally formed intestinal microflora, 60 percent had normal microflora in the fauces, 80 percent in the nose, and 70 percent of the neonates demonstrated normal skin microflora . The carriage of opportunistic microorganisms in the fauces and nose of the neonates had decreased substantially by the time of the discharge from the maternity home, while the incidence of pyo-inflammatory diseases had also reduced among them. J Rheumatol, 1989 Jan, 16(1), 92 - 6 Serum hyaluronate as a marker for disease severity in the Lactobacillus casei model of arthritis in the rat; Goldberg RL et al.; Serum concentrations of hyaluronate may provide a clinically relevant, quantitative marker of disease in patients with active rheumatoid arthritis (RA) . We studied the utility of serum hyaluronate in an animal model, with features reminiscent of human RA, in which LEW/N female rats were made arthritic by intraperitoneal injection of sonicated Lactobacillus casei . When serum hyaluronate was measured by an inhibition ELISA, a dose dependent correlation was found between the amount of L . casei injected and both joint score and serum hyaluronate in the chronic phase of the disease . A linear correlation between the chronic phase joint score and serum hyaluronate was observed (r = 0.69, p less than 0.001) . Two orally administered compounds, flurbiprofen (20 mg/kg) and methotrexate (0.125 mg/kg), were effective in decreasing both variables . Thus, serum hyaluronate may have utility in evaluating the therapeutic efficacy of antirheumatic/antiinflammatory agents in vivo in the chronic phase of RA-like diseases. Cancer Immunol Immunother, 1989, 28(4), 253 - 9 Role of culture supernatant of cytotoxic/cytostatic macrophages in activation of murine resident peritoneal macrophages; Hashimoto S et al.; Peritoneal macrophages elicited by Lactobacillus casei YIT9018 (LCEPM) were incubated in culture for 18 h with L . casei; the culture supernatant (LCM) was then harvested and tested for its ability to increase the cytostatic activity of resident peritoneal macrophages (RPM) and LCEPM . Treatment of RPM with LCM induced activation of macrophages to a cytostatic state against L929, Colon 26, P815, P388D1 and L1210 cells . A combination of recombinant human tumor necrosis factor (rhTNF), recombinant mouse TNF (rmTNF), recombinant human interleukin-1 (rhIL-1) or bacterial lipopolysaccharide with recombinant mouse interferon gamma (rmIFN-gamma) resulted in the synergistic induction of cytostatic activity in RPM . Recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) plus rhTNF increased the cytostatic activity of RPM a little but rmGM-CSF or rhTNF combined with rhIL-1 or alone had no effect . The effect of LCM on RPM was not inhibited by polymyxin B, anti-mTNF antiserum or below 20 U/ml monoclonal anti-rmIFN-gamma antibody (anti-rmIFN-gamma) but was inhibited by more than 40 U/ml anti-rmIFN-gamma, and LCM did not have any interferon antiviral activity . These results suggest that the cytostatic activity of RPM was augmented by the LCM, and that the effect of the LCM may be not due to IFN-gamma, TNF, GM-CSF, IL-1 or a small amount of contaminating lipopolysaccharide. J Bacteriol, 1989 Jan, 171(1), 280 - 4 Transport of amino acids in Lactobacillus casei by proton-motive-force-dependent and non-proton-motive-force-dependent mechanisms; Strobel HJ et al.; Lactobacillus casei 393 cells which were energized with glucose (pH 6.0) took up glutamine, asparagine, glutamate, aspartate, leucine, and phenylalanine . Little or no uptake of several essential amino acids (valine, isoleucine, arginine, cysteine, tyrosine, and tryptophan) was observed . Inhibition studies indicated that there were at least five amino acid carriers, for glutamine, asparagine, glutamate/aspartate, phenylalanine, or branched-chain amino acids . Transport activities had pH optima between 5.5 and 6.0, but all amino acid carriers showed significant activity even at pH 4.0 . Leucine and phenylalanine transport decreased markedly when the pH was increased to 7.5 . Inhibitors which decreased proton motive force (delta p) nearly eliminated leucine and phenylalanine uptake, and studies with de-energized cells and membrane vesicles showed that an artificial electrical potential (delta psi) of at least -100 mV was needed for rapid uptake . An artificial delta p was unable to drive glutamine, asparagine, or glutamate uptake, and transport of these amino acids was sensitive to a decline in intracellular pH . When intracellular pH was greater than 7.7, glutamine, asparagine, or glutamate was transported rapidly even though the proton motive force had been abolished by inhibitors. Biotherapy, 1989, 1(2), 89 - 95 Induction of resistance in mice against murine cytomegalovirus by cellular components of Lactobacillus casei; Ohashi T et al.; Recently, we reported that heat-killed Lactobacillus casei (LC) protected mice from murine cytomegalovirus (MCMV) infection by augmentation of natural killer (NK) cell activity . In the present study, we examined which components of LC cell induce the nonspecific resistance most effectively . Whole cell preparation of original LC, susceptible to bacteriophages SG-T and J1, was more effective than its mutants resistant to either bacteriophage . Although the activity of LC cells decreased upon fractionation, cell wall fractions were more active than cytoplasmic fractions . Glycoprotein (GP), a cell wall constituent, was a potent inducer of the resistance . The relative activity of cellular components to induce the resistance was evaluated by a protection index, a ratio of plaque-forming units (PFU) per 50% lethal dose (LD50) for treated mice to that for untreated mice . The protection indices of LC cells and GP were approximately 80 and 28, respectively . The protective effect of GP was evidenced by a decrease in titers of infectious viruses replicated in the target organs . Not only LC cells but also GP, although to a lesser degree, enhanced NK cell activity both in uninfected mice and MCMV-infected mice . The activity of LC cells and GP to augment NK cell activity correlated with the protection index . GP treatment did not modify interferon (IFN) production during MCMV infection . Thus, GP of LC cells seems to be the active principle to endow mice with resistance to MCMV. Lab Delo, 1989, (11), 59 - 61 {A method of determining the magnitude of the intracellular reserve of heparin in the blood}; Gabitov SZ et al.; The new method for assessment of the function of the blood anticoagulation system cellular component is based on the detection of the intracellular heparin amount that can be released into the blood stream . The technique involves the recording of changes in the blood plasma heparin levels after addition of lactobacterin, a nonspecific cell stimulant, to whole blood . The informative value of this method for the assessment of the blood anticoagulation system status is illustrated with examples. Zentralbl Mikrobiol, 1989, 144(1), 53 - 7 Occurrence of conjugative amylolytic activity in rumen lactobacilli; Kmet V et al.; 1,263 rumen strains of Lactobacilli were selected for the production of amylase and resistance to antibiotics . Out of 48 amylolytic Lactobacilli, in 17 strains, there was a cotransfer of the alpha amylase activity and resistance to tetracyclin with a transfer frequency of 10(-3) to 10(-7) ml-1 of the conjugative mixture . In strains No . 30, No . 36 and No . 49, plasmids with similar molecular weights as plasmids in corresponding transconjugants were detected . Similar protein pattern of the donors recipient of the L . lactis RN 1 and transconjugants obtained by PAGE proved the cotransfer of the amylase activity and antibiotic resistance. Int J Food Microbiol, 1988 Dec 31, 7(4), 339 - 47 Ropy slime formation in vacuum-packed cooked meat products caused by homofermentative lactobacilli and a Leuconostoc species; Korkeala H et al.; This paper deals with the cause of the formation of ropy slime on the surface of vacuum-packed cooked meat products . Two different homofermentative lactobacilli and a Leuconostoc strain were isolated from different ropy vacuum-packed meat products . The strains were able to produce slime experimentally on vacuum-packed cooked sausages . The slime had a molecular weight in the range of 70000-30000 and contained glucose and galactose in a ratio 10:1-10:2 . The formation of ropy slime also occurred on sausages without added sugars. Int J Food Microbiol, 1988 Dec 31, 7(4), 317 - 30 Identification of major contamination sources during processing of emulsion sausage; Borch E et al.; The extent of contamination of an emulsion type of sausage with lactic acid bacteria was determined along the processing line . This was done by aseptically removing sausages after five different processing stages (heat processing, chilling, cold storage, cutting down and packing) . Removed sausages were vacuum-packed and stored at 8 degrees C . The microbial growth was followed during storage and the microbiological shelf-life obtained at the different stages of the processing was determined . The spoilage flora of stored sausages was identified/grouped . Two major hygienic problems were identified: (1) a heat tolerant flora of Lactobacillus viridescens which survived the heat processing and was never outgrown by the recontaminating flora; (2) recontamination with a flora dominated by Lactobacillus sp . group 5, which occurred in the cold storage room; this flora dominated in the absence of L . viridescens . The heat tolerant L . viridescens SMRICC 193 survived at 68 degrees C for 40 min . Being exposed to a slowly increasing temperature, only a 10 cfu/ml decrease took place when the temperature increased from 60 degrees C to 70 degrees C over a period of 30 min. Int J Food Microbiol, 1988 Dec 31, 7(4), 299 - 309 Effect of environmental conditions during soya-bean fermentation on the growth of Staphylococcus aureus and production and thermal stability of enterotoxins A and B; Nout MJ et al.; During tempe manufacture, Staphylococcus aureus was able to survive in newly started soya-bean soaks (final pH 4.7), whereas it died during soaks with accelerated souring (final pH 4.0) . No S . aureus were detected after subsequent heat treatment of soaked beans . S . aureus strain S6 could produce enterotoxins A and B if inoculated on cooked soya-beans . Its growth and enterotoxin production were not influenced by inoculation rate nor by the presence of actively growing Rhizopus oligosporus, the fungus responsible for tempe formation . Enterotoxin levels were highest after 48 h fermentation . Presence of lactobacilli (L . brevis or L . plantarum) during the tempe fermentation did not influence staphylococcal growth, but enterotoxin levels we