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PubMed1980

Folate analogues altered in the C9-N10 bridge region. 16. Synthesis and antifolate activity of 11-thiohomoaminopterin.

The synthesis of 11-thiohomoaminopterin (1), which is a close analogue of 11-thiohomofolic acid (2), has been carried out by modification of the Boon-Leigh procedure. Treatment of 1-chloro-4-[p-(carbomethoxy)thiopenoxy]-2-butanone (5) with sodium azide gave 1-azido-4-[p-(carbomethoxy)thiophenoxy]-2-butanone (6). After protection of the carbonyl group of 6, the product 7 was catalytically hydrogenated to 1-amino-4-[p-(carbomethoxy)thiophenoxy]-2-butanone ketal (3). Reaction of 32 with 6-chloro-2,4-diaminmo-5-nitropyrimidine gave the desired pyrimidine intermediate, which was elaborated to 4-amino-4-deoxy-11-thiohomopteroic acid (20) by standard procedures. Alternately, 1-azido-4-[p-(carbomethoxy)thiophenoxy]-2-butanone ketal (7) was hydrolyzed to the corresponding acid (8) and coupled with diethyl L-glutamate to obtain diethyl N-[p-(1-azido-2-oxo-4-thiobutanoyl)benzoyl]-L-glutamate ketal (10), which was used for the large-scale preparation of 11-thiohomoaminopterin (1). Although 11-thiohomoaminopterin showed antifolate activity against two folate-requiring microorganisms and inhibited Lactobacillus casei dihydrogolate reductase, it did not exhibit any antitumor activity against L-1210 lymphoid leukemia in mice at a maximum dose of 48 mg/kg.

Animals↗
PubMed1999

Study of adhesion of Lactobacillus casei CRL 431 to ileal intestinal cells of mice.

It is well known that the cell wall of Lactobacillus casei CRL 431, a strain present in probiotics, presents lectinlike surface molecules. Presence of these molecules stimulates the immune system. Given the role that lectins and lectinlike substances play in the adhesion phenomenon, it is probable that this is an initial stage in the immunostimulation produced by this bacterium. To confirm this, adhesion of this microorganism to exfoliated mouse ileal epithelial cells was studied in vitro. Other L. casei strains isolated from adult human intestines and one of dairy origin were also examined for their ability to adhere to ileal epithelial cells. Another strain, which was included in the present study, was Lactobacillus acidophilus CRL 730. L. casei strains isolated from humans showed good ability to adhere to ileal epithelial cells, whereas L. casei isolated from dairy origin did not. Adhesion was only observed at 37 degrees C and at a pH between 6 and 7.5. The exposure time needed for highest adhesion was 30 min. Presence of lectinlike substances on the surface of L. casei CRL 431 is important to this adhesion phenomenon, since adherence capacity was lost after removal of these substances.

Animals↗
PubMed2006

Effects of sonicated Prevotella intermedia, Fusobacterium nucleatum and Lactobacillus casei extracts on interleukin-8 production by human dental pulp cells.

The objective of this study was to determine the effects of Prevotella intermedia, Fusobacterium nucleatum and Lactobacillus casei on the production of IL-8 by human dental pulp cells. Human dental pulp cells from teeth of young patients (aged 18-25 years) were cultured and tested with sonicated P. intermedia ATCC 25611, F. nucleatum ATCC 25586 and L. casei ATCC 4646 extracts. IL-8 secreted into the culture supernatants were measured at 6, 12 and 24 hours using a quantitative sandwich enzyme immunoassay technique. Cell viability was evaluated using trypan blue exclusion technique. IL-8 production by human dental pulp cells increased significantly at 12 and 24 hours after exposure to P. intermedia and F. nucleatum, whereas L. casei extract exhibited low IL-8 production. The sonicated bacterial extracts did not significantly affect viability or total number of dental pulp cells.

Adolescent↗
PubMed2005

Lactic acid bacteria biomass monitoring in highly conductive media by permittivity measurements.

Recent technological improvements have extended the application range of permittivity biomass measurements to bacterial fermentations in highly conductive media. With Lactobacillus casei, the effective biomass detection sensitivity of the FOGALE Biomass System is around 0.2 g/l (0.01 pF/cm). Fermentations growth kinetics of Lactobacillus casei can be recorded with good reproducibility and accuracy despite the high medium conductivity varying between 15 and 75 mS/cm, and the low cell concentration (<6 g/l).

Biomass↗
PubMed2001

Characterization of the Lactobacillus casei group and the Lactobacillus acidophilus group by automated ribotyping.

A total of 91 type and reference strains of the Lactobacillus casei group and the L acidophilus group were characterized by the automated ribotyping device Riboprinter microbial characterization system. The L. casei group was divided into five (C1-C5) genotypes by ribotyping. Among them, the strain of L. casei ATCC 334 was clustered to the same genotype group as most of L. paracasei strains and L casei JCM 1134T generated a riboprint pattern that was different from the type strain of L. zeae. These results supported the designation of L. casei ATCC 334 as the neotype strain, but were not consistent with the reclassification of L. casei JCM 1134T as L. zeae. The L. acidophilus group was also divided into 14 (A1-A11, B1-B3) genotypes by ribotyping. L. acidophilus, L. amylovorus, L. crispatus and L. gallinarum generated ribotype patterns that were distinct from the patterns produced by L. gasseri and L. johnsonii. This result confirmed previous data that the L. acidophilus group divided to two major clusters. Five strains of L. acidophilus and two strains of L. gasseri were correctly reidentified by ribotyping. Most strains belonging to the L. casei group and the L. acidophilus group were discriminated at the species level by automated ribotyping. Thus this RiboPrinter system yields rapid, accurate and reproducible genetic information for the identification of many strains.

Humans↗
PubMed1985

Characterization and molecular cloning of cryptic plasmids isolated from Lactobacillus casei.

Four small cryptic plasmids were isolated from Lactobacillus casei strains, and restriction endonuclease maps of these plasmids were constructed. Three of the small plasmids (pLZ18C, pLZ19E, and pLZ19F1; 6.4, 4.9, and 4.8 kilobase pairs, respectively) were cloned into Escherichia coli K-12 by using pBR322, pACYC184, and pUC8 as vectors. Two of the plasmids, pLZ18C and pLZ19E, were also cloned into Streptococcus sanguis by using pVA1 as the vector. Hybridization by using nick-translated cloned 32P-labeled L. casei plasmid DNA as the probe revealed that none of the cryptic plasmids had appreciable DNA-DNA homology with the large lactose plasmids found in the L. casei strains, with chromosomal DNAs isolated from these strains. Partial homology was detected among several plasmids isolated from different strains, but not among cryptic plasmids isolated from the same strain.

Chromosome Mapping↗
PubMed2004

Initiation of protein synthesis by a labeled derivative of the Lactobacillus casei DN-114 001 strain during transit from the stomach to the cecum in mice harboring human microbiota.

Although studies on the survival of bacteria in the digestive tract have been reported in the literature, little data are available on the physiological adaptation of probiotics to the digestive environment. In previous work, a transcriptional fusion system (i.e., luciferase genes under the control of a deregulated promoter) was used to demonstrate that a derivative of the Lactobacillus casei DN-114 001 strain, ingested in a fermented milk and thus exhibiting initially a very weak metabolic activity, synthesized proteins de novo after its transit in the digestive tract of mice harboring human microbiota (known as human-microbiota-associated mice). With the same genetic system and animal model, we here investigate for the first time the ability of L. casei to reinitiate synthesis in the different digestive tract compartments. In this study, most ingested L. casei cells transited from the stomach to the duodenum-jejunum within 1 h postingestion. No luciferase activity was observed in these digestive tract compartments after the first hour. At later times, the bulk of bacteria had transited to the ileum and the cecum. Luciferase synthesis was detected between 1.5 and 2.0 h postingestion at the ileal level and from 1.5 h to at least 6.0 h postingestion in the cecum, where the activity remained at a maximum level. These results demonstrate that ingested L. casei (derivative of the DN-114 001 strain) administered via a fermented milk has already reinitiated protein synthesis when it reaches the ileal and cecal compartments.

Animals↗
PubMed1999

Effect of applying lactic acid bacteria isolated from forage crops on fermentation characteristics and aerobic deterioration of silage.

Two selected strains, Lactobacillus casei FG 1 and Lactobacillus plantarum FG 10 that were isolated from forage crops were used as additives at 1.0 x 10(5) cfu/g of fresh matter to alfalfa, Italian ryegrass, and sorghum, and their effect on fermentation characteristics and aerobic deterioration of silage was studied. The three silages treated with strains FG 1 or FG 10 were well preserved; had significantly lower pH values, butyric acid, propionic acid, and ammonia N concentrations, gas production, and dry matter losses; and had significantly higher contents of residual water-soluble carbohydrates and lactic acid than did the respective control silages. Yeast counts were high in all treated silages and increased rapidly during aerobic exposure. As a result, treated silages spoiled faster upon aerobic exposure than did the respective control silages. Most yeasts isolated from deteriorated silages showed high tolerance to lactic acid but low tolerance to butyric acid, and they were able to grow at low pH conditions and assimilate lactic acid. The results confirmed that L. casei and L. plantarum improved fermentation quality but did not inhibit the growth of silage yeast or aerobic deterioration of the silage.

Aerobiosis↗
PubMed2005

The glycolytic genes pfk and pyk from Lactobacillus casei are induced by sugars transported by the phosphoenolpyruvate:sugar phosphotransferase system and repressed by CcpA.

In Lactobacillus casei BL23, phosphofructokinase activity was higher in cells utilizing sugars transported by the phosphoenolpyruvate:sugar phosphotransferase system (PTS). The phosphofructokinase gene (pfk) was cloned from L. casei and shown to be clustered with the gene encoding pyruvate kinase (pyk). pfk and pyk genes are cotranscribed and induced upon growth on sugars transported by the PTS. Contrarily to the model proposed for Lactococcus lactis, where the global catabolite regulator protein (CcpA) is involved in PTS-induced transcription of pfk and pyk, a ccpA mutation resulted in a slight increase in pfk-pyk expression in L. casei. This weak regulation was evidenced by CcpA binding to a region of the pfk-pyk promoter which contained two cre sequences significantly deviated from the consensus. The PTS induction of pfk-pyk seems to be counteracted by the CcpA-mediated repression. Our results suggest that the need to accommodate the levels of pfk-pyk mRNA to the availability of sugars is fulfilled in L. casei by a PTS/CcpA-mediated signal transduction different from L. lactis.

Amino Acid Sequence↗
PubMed2005

The treatment of mice with Lactobacillus casei induces protection against Babesia microti infection.

In this study, we report that administration of Lactobacillus casei confers protection to mice against the intracellular protozoan Babesia microti. Mice treated with L. casei orally or intraperitoneally were inoculated 7 days later with an infectious dose of B. microti. Mice treated with lactobacilli showed significant reduction in the percentage of parasitized erythrocytes (PPE) compared to untreated mice. When mice were inoculated intraperitoneally with L. casei 3 or 0 days before challenge with B. microti, the PPE was significantly lower compared to untreated mice and there were no differences between treated mice and mice immune to B. microti infection. When mice treated with live or dead L. casei were compared to mice inoculated with Freund Complete Adjuvant before a B. microti infection, a significant reduction of PPE was observed. These results show the protective effect of L. casei administered to mice against a B. microti infection and suggest that it might act by stimulating the innate immune system.

Administration, Oral↗
PubMed2005

In vitro antibacterial effects of the dentin primer of Clearfil Protect Bond.

OBJECTIVES: This study aimed to investigate the antibacterial effects of the dentin primer of a commercially available self-etching adhesive system, Clearfil Protect Bond, which contains antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB). METHODS: Inhibitory effects against Streptococcus mutans, Lactobacillus casei, or Actinomyces naeslundii were examined by an agar-disc diffusion method using the Clearfil Protect Bond primer containing 5% MDPB and an acidic adhesion-promoting monomer MDP, the primer only with MDP, and the primer with 1% cetylpyridinium chloride. The minimum inhibitory/bactericidal concentrations (MIC/MBC) of each primer for the three bacterial species were determined by serial microdilution assays. For testing the bactericidal effects seen in dentin, the primer was applied to demineralized dentin blocks in which S. mutans had been impregnated, and numbers of viable bacteria were counted. RESULTS: For all three bacteria, the sizes of the inhibition zones produced by Clearfil Protect Bond primer were significantly greater than for the other primers (p<0.05, ANOVA and Scheffe's F-test). The MIC/MBC values of Clearfil Protect Bond primer were less than those of the primer without MDPB, and comparable to those of the primer containing cetylpyridinium chloride. No bacterial recovery was obtained after application of Clearfil Protect Bond primer to the bacteria-impregnated dentin, although the primer without MDPB showed some bactericidal effect. SIGNIFICANCE: Clearfil Protect Bond primer has strong antibacterial activity based upon MDPB against S. mutans, L. casei and A. naeslundii, and the capability to disinfect cavities containing residual bacteria.

Acid Etching, Dental↗
PubMed2001

Effect of viable or dead Lactobacillus casei organisms administered orally to mice on resistance against Trichinella spiralis infection.

The capacity of viable, dead Lactobacillus casei and supernatant from L. casei culture, administered by oral route, to induce resistance in mice against Trichinella spiralis infection was evaluated. The percentage of adult worm reduction in the intestine five days after T. spiralis infection as compared with the worm burden in the control group fluctuated between between 53.1 and 58% in mice treated with viable L. casei, while reductions in animals treated with dead lactobacilli or supernatant from L. casei culture were of 44 and 32.5% respectively. The percentage of larvae per gram of muscle tissue reductions, as compared with controls, obtained 30 days after infection ranged from 48.4 to 70.7% in rodents which ingested viable L. casei compared with the percentage reductions of 65.9 and 24% obtained respectively in mice treated with dead lactobacilli or with L. casei supernatant. The protective response observed in the present study may be explained on the basis of 1) lactobacilli colonization of the intestine, 2) macrophage processing of dead lactobacilli in local immune tissues and presentation of L. casei antigens to Th1 cells which, in turn, produced IL-2 to activate B cells and other T cells.

Administration, Oral↗
PubMed1977

Urinary riboflavin metabolites in the rat.

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

Age Factors↗
PubMed1997

The association of yogurt starters with Lactobacillus casei DN 114.001 in fermented milk alters the composition and metabolism of intestinal microflora in germ-free rats and in human flora-associated rats.

The aim of this study was to compare the effects of milk and of various fermented milks on the composition and metabolic activities of the intestinal microflora. Groups of eight rats were fed for 6 wk a diet containing 30% nonfermented milk (M), yogurt (Y), milk fermented with Lactobacillus casei (LcFM) or milk fermented with the association of L. casei DN 114.001 and yogurt starters (LcYFM). In the first study, the survival of the lactic acid bacteria from the fermented milks was assessed by bacterial enumeration in feces of germ-free rats (GF rats) fed milk or fermented milks. The metabolic activities of the lactic acid bacteria were studied in these rats by the measurement of glycolytic activities and products of bacterial fermentation, i.e., acetate and lactate (isoforms L and D). In a second study, the effects of fermented milks on the composition and metabolism [gas, glycolytic activities, short-chain fatty acids (SCFA), alcohol and ammonia] of human flora were studied using human flora-associated rats (HF rats). In GF rats, the survival of L. casei in the feces did not differ between those fed the LcFM and LcYFM diets. L. bulgaricus was detected in the feces of the rats fed Y, whereas Streptoccus thermophilus was found in the feces of the LcYFM group. In HF rats, fecal concentration of Bifidobacteria was greater in the LcFM group than in the others. beta-Glucuronidase (EC 3.2.1.31) activity was lower in rats fed LcFM and Y than in those fed M and LcYFM, whereas beta-galactosidase (3.2.1.23), alpha-glucosidase (EC 3.2.1 20) and beta-glucosidase (EC 3.2.1.21) activities were higher in the LcYFM group compared with the others. Methane excretion was higher in rats fed Y than in other groups. Cecal SCFA concentrations did not differ in LcFM, Y and M groups, but total SCFA, acetate, propionate and butyrate were significantly greater in the LcYFM group. These results suggest that milk fermented with the combination of L. casei and yogurt starters leads to specific effects that are different from the simple addition of the effects found with yogurt and milk fermented with L. casei. These specific effects are potentially beneficial to human health.

Acetates↗
PubMed1993

Diet during rotavirus enteritis affects jejunal permeability to macromolecules in suckling rats.

We studied the influence of diet during diarrhea on gut mucosal barrier in a suckling rat model. Rat pups were inoculated with IDIR virus (a group B rotavirus) at 10 d of age. Beginning 2 d postinfection, in addition to maternal milk, group CM received a daily gavage of cow milk and group GG received Lactobacillus casei strain GG, a human strain previously shown to survive the passage through the gastrointestinal tract and temporarily colonize the gut. Group CMGG received a combination of these, and control animals were gavaged with tap water. At 21 d of age, jejunal absorption of intact and degraded horseradish peroxidase (HRP) in Ussing chamber was markedly higher in IDIR virus-infected than in noninfected controls. In the two groups gavaged with cow milk, group CM and group CMGG, the numbers of specific antibody-secreting cells (enumerated by the solid-phase enzyme-linked immunospot assay) against beta-lactoglobulin were significantly higher than in the groups that had not received cow milk. In parallel with immune system activation, a statistically significant increase in the absorption of intact HRP (mean and 95% confidence interval, ng x h-1 x cm-2) was detected: group CM, 302 (155, 586); group CMGG, 174 (56, 545); infected controls, 121 (57, 257); and group GG, 44 (8, 254). A decrease in the uptake of intact HRP (F = 3.64, p = 0.06) and degraded HRP (F = 9.50, p = 0.004) was associated with the introduction of L. casei GG to the diet, irrespective of coexposure to cow milk.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2003

Effect of Lactobacillus strains and Saccharomyces boulardii on persistent diarrhea in children.

The efficacy of probiotics on persistent diarrhea remains uncertain. The purpose of this study was to evaluate the effect of Lactobacillus sp and Saccharomyces boulardii on persistent diarrhea in children. In a double-blind trial eighty-nine children, aged 6-24 months were randomly distributed to receive pasteurized cow milk containing 2 viable lyophilized strains Lactobacillus casei and Lactobacillus acidophillus strains CERELA, (10(10)-10(12) colony-forming units per g) (n = 30), or lyophilized S. boulardii, (10(10)-10(12) colony forming units per g) (n = 30) or pasteurized cow milk as placebo (n = 29); on each diet 175 g was given twice a day for a 5 day period. Number of depositions, duration of illness and frequency of vomiting were considered. Enteric pathogens were isolated from stools in 40% of the patients, 27% had rotavirus. Lactobacillus and S. boulardii significantly reduced the number of depositions (p < 0.001) and diarrheal duration (p < 0.005). Similarly both significantly (p < 0.002) reduced vomiting as compared with placebo. There was no difference between treatments depending on rotavirus status. In conclusion, L. casei and L. acidophillus strains CERELA and S. boulardii are useful in the management of persistent diarrhea in children.

Child, Preschool↗
PubMed2004

Lactobacillus casei, dominant species in naturally fermented Sicilian green olives.

This study investigated the phenotypic and genotypic characteristics of lactic acid bacteria in naturally fermented green olives, collected from different areas of Sicily. Both classical biochemical tests and PCR/Restriction Fragments Length Polymorphism (RFLP) of 16S rDNA were used to characterize the isolates. The identity of the isolates was obtained by the partial sequencing analysis of the 16S rDNA. The BioMerieux software assigned the 13 heterofermentative strains to the Lactobacillus brevis species; 24 homofermentative strains were classified as Lactobacillus casei and the remaining 11 homofermentative lactobacilli were identified as Lactobacillus plantarum. The rapid ID 32 STREP test identified coccal-shaped strains as Enterococcus faecium species. The PCR/RFLP analysis showed a remarkable bacterial heterogeneity within the isolates. The 16S rDNA partial sequencing did not confirm biochemical identification, revealing a strong dominance of isolates belonging to the L. casei species. It is noteworthy that this species has never been reported as dominant species in fermented vegetables.A combination of molecular and biochemical analysis allowed the identification of species involved in natural food fermentations.

Fermentation↗