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Induction by a lactic acid bacterium of a population of CD4(+) T cells with low proliferative capacity that produce transforming growth factor beta and interleukin-10.

We investigated whether certain strains of lactic acid bacteria (LAB) could antagonize specific T-helper functions in vitro and thus have the potential to prevent inflammatory intestinal immunopathologies. All strains tested induced various levels of both interleukin-12 (IL-12) and IL-10 in murine splenocytes. In particular, Lactobacillus paracasei (strain NCC2461) induced the highest levels of these cytokines. Since IL-12 and IL-10 have the potential to induce and suppress Th1 functions, respectively, we addressed the impact of this bacterium on the outcome of CD4(+) T-cell differentiation. For this purpose, bacteria were added to mixed lymphocyte cultures where CD4(+) T-cells from naive BALB/c mice were stimulated weekly in the presence of irradiated allogeneic splenocytes. In these cultures, L. paracasei NCC2461 strongly inhibited the proliferative activity of CD4(+) T cells in a dose-dependent fashion. This was accompanied by a marked decrease of both Th1 and Th2 effector cytokines, including gamma interferon, IL-4, and IL-5. In contrast, IL-10 was maintained and transforming growth factor beta (TGF-beta) was markedly induced in a dose-dependent manner. The bacteria were not cytotoxic, because cell viability was not affected after two rounds of stimulation. Thus, unidentified bacterial components from L. paracasei NCC2461 induced the development of a population of CD4(+) T cells with low proliferative capacity that produced TGF-beta and IL-10, reminiscent of previously described subsets of regulatory cells implicated in oral tolerance and gut homeostasis.

Animals↗

Influence of the stabilizer coating layer on the purification and freeze-drying of poly(D,L-lactic acid) nanoparticles prepared by an emulsion-diffusion technique.

In this study, the purification by cross-flow filtration (CFF) and freeze drying of poly(D,L-lactic acid) (PLA) nanoparticles prepared by an emulsion-diffusion technique using poly(vinyl alcohol) (PVAL) or poloxamer 188 (P-188) were investigated. The stability of the suspensions was correlated to the affinity of the stabilizers for the nanoparticle surface, the resistance of the coating layer to continuous filtration and to freeze-thawing procedures. The results indicated a clear difference between the two stabilizers, suggesting that the nature of the coating layer has a very important role during CFF and freeze-drying. Nanoparticles prepared with PVAL were filtered and freeze-dried without nanoparticle fusion. This behaviour was attributed to the formation of a stable thick layer (similar to that found for polystyrene latex). In contrast, aggregation of nanoparticles was observed during CFF for the batches prepared with P-188, indicating that the polypropylene oxide blocks present in the copolymer have little affinity for the PLA surface. However, these suspensions were successfully recovered when using stabilizer solutions as diafiltration media, suggesting a dynamic exchange between the P-188-adsorbed chains and those of the identical polymer remaining in the bulk solution. The presence of P-188 did not prevent nanoparticle aggregation after freeze-drying. Therefore, the use of cryoprotectants was necessary. Aggregation may have been due to an increase in the solubility of P-188 in the bulk solution, which provokes a destabilization of the suspension by desorption and partial coverage of the surface. The best cryoprotectants were found to be sugars containing glucose units. The cryoprotective effect was related to the hydrogen bonding capability of these sugars, which prevented aggregation by dehydration of P-188 forcing it to the PLA surface.

Chemistry, Pharmaceutical↗

Organotin(IV) compounds as intramolecular transesterification catalysts in thermal depolymerization of poly(L-lactic acid) oligomer to form LL-lactide.

Mono-, di-, and tetraalkyl tin(IV) compounds were evaluated for the intramolecular transesterification reaction of the thermal depolymerization of poly(L-lactic acid) oligomer forming lactide by gas chromatography using a beta-cyclodextrin stationary phase capillary column. The most active catalyst was found to be monobutyltin trichloride (BuSnCl3) (8), which contains tin-halogen bonds, and the least effective was the coordinatively saturated monoorganotin derivative, monobutyltin tris(2-ethylhexanoate) (7). Coordination of the carbonyl group in the oligomer to the tin catalysts is an important factor influencing its activity.

Catalysis↗

[Corrosiveness of Ti-Fe-Mo-Mn-Nb-Zr alloys in various pH lactic acids].

Variation in pH value of oral cavity circumstance causes decayed tooth. In this paper, corrosions of Ti-Fe-Mo-Mn-Nb-Zr dental alloys in various pH lactic acids were studied by immersion test. In the case of pH=4, which is an acidity slightly larger than that causes decayed tooth, Ti-Fe-Mo-Mn-Nb-Zr alloys are entirely corrosion-resistant, and the corrosion type is pitting. With the increase in acidity, the corrosion-resistance of Ti-Fe-Mo-Mn-Nb-Zr alloys deteriorates, and the corrosion type changes from pitting to intergranular corrosion. Fe ion is the most dissolved substance in impregnation, which means Fe goes against the corrosion-resistance of Ti alloys. Both results of thermodynamics calculation and X-ray photoelectron spectroscopy have shown that Mn2O3, Nb2O5 and TiO2 form oxide film on the surface of Ti-Fe-Mo-Mn-Nb-Zr alloys.

Corrosion↗

Behavior of variable V3 region from 16S rDNA of lactic acid bacteria in denaturing gradient gel electrophoresis.

Separation of amplified V3 region from 16S rDNA by denaturing gradient gel electrophoresis (DGGE) was tested as a tool for differentiation of lactic acid bacteria commonly isolated from food. Variable V3 regions of 21 reference strains and 34 wild strains referred to species belonging to the genera Pediococcus, Enterococcus, Lactococcus, Lactobacillus, Leuconostoc, Weissella, and Streptococcus were analyzed. DGGE profiles obtained were species-specific for most of the cultures tested. Moreover, it was possible to group the remaining LAB reference strains according to the migration of their 16S V3 region in the denaturing gel. The results are discussed with reference to their potential in the analysis of LAB communities in food, besides shedding light on taxonomic aspects.

DNA, Bacterial↗

[Hydrolysis of poly(L-lactic acid) fibers and formation of low crystalline apatite on their surface by a biomimetic process].

Low crystalline apatite coating was formed on the surface of biodegradable poly(L-lactic acid) (PLLA) fibers by a biomimetic process, i.e., by immersing the fibers in a modified simulated body fluid (SBF) at 37 degrees C and pH 7.3 after hydrolysis of the fibers in water. The apatite was characterized by scanning electron microscopy with energy dispersive X-ray spectroscopy, X-ray photoelectron spectroscopy, X-ray diffraction and Fourier transform infrared spectroscopy. Results showed that the fiber hydrolysis could accelerate the apatite formation but had little effect on the chemical and crystalline structure of the apatite. The structure of the apatite coating formed by the biomimetic method was similar to that of apatite in the natural bone. The bone-like low crystalline apatite coating might exhibit enhanced osteo-conductivity when the PLLA fibers are applied in bone reconstruction biomaterials.

Apatites↗

A liquid-stable reagent for lactic acid levels. Application to the Hitachi 911 and Beckman CX7.

We evaluated the use of a new lactate oxidase-based reagent for the determination of serum and plasma lactic acid levels with the Hitachi 911 (Roche Diagnostics, Indianapolis, IN) and the Beckman CX7 (Beckman Instruments, Brea, CA). Evaluation studies demonstrated on-board stability of at least 3 months and a calibration stability of more than 5 months. Within- and between-day imprecision of this reagent was less than 2% for both applications. The reagent is free of the deleterious effects of triglyceride up to levels of 1,400 mg/dL (15.8 mmol/L), bilirubin to concentrations of 24.6 mg/dL (420 mumol/L), and hemoglobin, from lysed erythrocytes, to levels of more than 0.3 g/dL (3.0 g/L). When used on the Hitachi 911 for the determination of plasma lactate concentrations, the reagent correlates with the Dade aca III (Dade International, Deerfield, IL). When applied to the Beckman CX7 for the determination of serum lactate levels, the method correlates with the Beckman method.

Ampyrone↗

Effect of chosen lactic acid bacteria strains on Staphylococcus aureus in vitro as well as in meat and raw sausages.

The present study was aimed at determining the influence of 15 strains of lactic acid bacteria on the growth of 2 Staphylococcus aureus strains in vitro as well as in meat and raw sausages. The investigations were performed within the framework of three alternate stages which differed in respect to the products studied, the number of Lactobacillus sp. strains and, partly, methodological approach. The study also considered water activity (a(w)) and pH of the products investigated. The results obtained are demonstrated in 5 diagrams. It was found that among 15 strains of Lactobacillus aureus investigated only one strain, Lactobacillus helveticus T 78, showed antagonistic effect on studied strains of Staphylococcus aureus both in vitro as well as in meat and raw sausages. Five other strains of Lactobacillus spp. displayed the antagonistic effect in vitro only. The temperature and incubation time of sausages, but also the type of sausage stuffing were found to have a distinct or slight influence, respectively, on the antagonistic interaction between the bacteria. However, this phenomenon was affected by neither a(w) nor pH.

Animals↗

Conjugates of poly(DL-lactic acid) with ethylenediamino or diethylenetriamino bridged bis(beta-cyclodextrin)s and their nanoparticles as protein delivery systems.

Some biodegradable amphiphilic copolymers were synthesized by conjugating poly(DL-lactic acid) (PLA) onto ethylenediamino or diethylenetriamino bridged bis(beta-cyclodextrin)s (bis-CDs). Double emulsion (DE) and nanoprecipitation (NP) methods were used to fabricate the nanoparticles of these copolymers entrapping bovine serum albumin (BSA) as a model protein. Effects of the experimental parameters, such as copolymer composition, BSA concentration, copolymer concentration and poly(vinyl alcohol) concentration, on particular size and encapsulation efficiency (EE) were investigated. Their EE to BSA could reach 83.5% at an optimized condition owing to the cooperative binding effect of the CD moiety with BSA. The core-corona structure of copolymer micelles fabricated from the nanoprecipitation was studied on the basis of 1H NMR and other measurements at various temperatures. The results showed that the core-corona structure kept stable below 50 degrees C (lower than Tg). And increase of the micelle association number occurred above the Tg because the size of the NPs became larger and proton signals of the liquid-like PLA cores could be observed in 1H NMR in D2O at 60 degrees C. The release profiles of NPs showed a burst effect followed by a continuous release. Sodium dodecyl sulfate polyacrylamide gel electrophoresis, circular dichroic and fluorescence spectra were further used to identify the stability of BSA released from the NPs. The nanoparticles from the conjugates have a promising potential in nasal delivery system.

Animals↗

Formulation factors affecting drug release from poly(lactic acid) (PLA) microcapsule tablets.

A sustained-release (SR) formulation of phenobarbital (PB) microcapsule tablet was prepared using low molecular weight (MW) DL- and high MW L-poly(lactic acid) (PLA) polymer. Microencapsulation of PB showed a unimodal size distribution (375 to 550 microns) of the microcapsules with high loading capacity (> 84%). Drug release from the microcapsule was influenced by the polymer ratios and increased with an increase in L-PLA amount. Microcapsules and physical mixtures of PB and the PLA were directly compressed independently to form microcapsule and matrix tablets, respectively. Drug release from the microcapsule tablets was significantly lowered (p < .001) compared to matrix tablets or free microcapsule (free microcapsule > matrices > microcapsule tablets). We also investigated the effect of tablet adjuvants, compression pressures, and microcapsule loading on the tablet performance in terms of friability, hardness, porosity, tensile strength, and the release kinetics of PB. The drug release rate increased with increasing compression pressure in the case of Emcompress or lactose, but not Avicel. The drug release rate was three- to fivefold increased with sodium starch glycolate compared to tablets without a disintegrant. With an increase in microcapsule loading, a decrease in the drug release rate was observed; however, the tablet performance remained satisfactory. The morphology of the microcapsules was monitored microscopically after the dissolution and the disintegration of tablets. The drug release accelerated with compression pressures and microcapsule loading from the tablets due to mechanical destruction of the microcapsule wall, which was more clearly seen after disintegration and dissolution of the tablets. Our data suggest that the PLA microcapsule can be tableted to make a SR product without significantly affecting its release kinetics.

Capsules↗

Paclitaxel loaded poly(L-lactic acid) microspheres for the prevention of intraperitoneal carcinomatosis after a surgical repair and tumor cell spill.

A controlled release delivery system for paclitaxel was developed using poly(L-lactic acid) to provide local delivery to the peritoneal cavity. Microspheres were made in 1-40 and 30-120 microm size ranges. In an in vitro release study, 30-120 microm microspheres loaded with 10, 20 and 30% paclitaxel exhibited a burst phase of release for 3 days followed by an apparently zero-order phase of release. At all loadings, 20-25% of the original load of paclitaxel was released after 30 days. The effect of microsphere size on retention in the peritoneal cavity was assessed. Control 1-40 microm microspheres were injected intraperitoneally in rats. The rats received either insufflation of the peritoneal cavity using 11 mmHg CO2 or no further treatment. After sacrifice, microspheres with diameters less than 24 microm were observed in the lymphatic system after being cleared from the peritoneal cavity through fenestrations in the diaphragm. Insufflation of the peritoneal cavity had no effect on the size of microspheres that were cleared. Efficacy studies were carried out using 30-120 microm microspheres that were of sufficient size to be retained in the peritoneal cavity. In a model of a tumor cell spill after a cecotomy repair, 100 mg of 30-120 microm microspheres containing 30% paclitaxel were effective in preventing growth of tumors in the peritoneal cavity at both 2 and 6 weeks post-surgery. No gross or histologically evident tumor growth was observed on any peritoneal surfaces or in the surgical wound site. Rats receiving control microspheres all showed tumor cell implantation and growth after 2 weeks.

Animals↗

Degradation of insoluble bovine collagen and human dentine collagen pretreated in vitro with lactic acid, pH 4.0 and 5.5.

The purpose of the study was to test the hypothesis that both insoluble pure type I collagen from bovine Achilles tendon and dentine collagen in root dentine powder from human teeth required acid pretreatment for subsequent degradation by trypsin, a non-specific protease. Pure type I collagen or dentine powder was treated with lactic acid, at pH 4 or 5.5, or distilled, deionized water (pH 7) as a negative control. After incubation at 37 degrees C for 24 h, extracts of pure type I collagen solutions were analysed for soluble collagen with the hydroxyproline assay. Extracts of dentine powder solution were analysed for Ca2+, total protein, final pH, and hydroxyproline. Residual, undegraded pellets were washed and then treated with trypsin or collagenase. After 24 h of incubation, the soluble fractions from the enzyme-treated pure type I collagen and dentine powder solutions were analysed for hydroxyproline. Results showed that almost no pure type I collagen was degraded during acid pretreatment. Trypsin degraded significantly more pure type I collagen in the pH 4-treated group than in the other groups. Collagenase degraded about 70% of the pure type I collagen irrespective of acid pretreatment. While acid pretreatment at pH 4 did not degrade dentine collagen, data from Ca2+ analyses and collagen breakdown by trypsin suggested that pretreatment at pH 4 demineralized and denatured dentine collagen so that the collagen could be subsequently degraded by enzymes. After pretreatment at pH 4, about 27 and 57% of the dentine collagen was degraded by trypsin and collagenase, respectively, in contrast to minimal degradation of non-acid-treated dentine collagen by the same enzymes.(ABSTRACT TRUNCATED AT 250 WORDS)

Achilles Tendon↗

Nucleoside-related mitochondrial toxicity among HIV-infected patients receiving antiretroviral therapy: insights from the evaluation of venous lactic acid and peripheral blood mitochondrial DNA.

Nucleoside analogues inhibit human DNA polymerase gamma. As a result, they can produce mitochondrial toxicity. We evaluated the possible role of random venous lactic-acid determinations as a screening tool for mitochondrial toxicity among patients receiving nucleoside therapy. More recently, we have developed an assay that can detect changes in mitochondrial DNA (mtDNA) levels in peripheral blood cells. Using this assay, we have characterized changes in mtDNA relative to nuclear DNA (nDNA) in peripheral blood cells from patients with symptomatic nucleoside-induced hyperlactatemia. Our results demonstrated that symptomatic hyperlactatemia was associated with markedly low mtDNA : nDNA ratios. A statistically significant increase in the mtDNA : nDNA ratio was observed after the discontinuation of antiretroviral therapy. Full validation of monitoring the mtDNA : nDNA ratio is currently under way.

Anti-HIV Agents↗

Factors affecting the ability of dental cements to alter the pH of lactic acid solutions.

Two related studies have been carried out to determine the effect of (a) powder:liquid ratio and (b) relative amounts of cement on the extent of buffering of a lactic acid storage solution using zinc polycarboxylate and glass-ionomer cements (both water-activated). The effect of varying the powder:liquid ratio was found to be slight and not statistically significant. On the other hand, increasing the numbers of specimens in a given volume of storage solution was found to have a significant influence on the final pH (at the 0.0005 level of significance). The fact that powder:liquid ratio did not affect the final pH led to the conclusion that attack occurs mainly at the matrix of these cements, confirming previous findings about the acid erosion process. The finding that increasing the relative amount of cement to acid storage solution strongly influenced pH suggested that, under clinical conditions, buffering would be important. When five specimens were used, the final pH corresponded to that of arrested caries, from which it was concluded that these cements have the ability to be cariostatic in vivo.

Acrylic Resins↗

Lactic acid removal and heart rate frequencies during recovery after strenuous rowing exercise.

Three tests were conducted to assess the effectiveness of three different intensities of exercise both in reducing blood lactic acid (LA) levels and in influencing subjects' heart rate (HR), following a 2000 m race in a rowing boat. In the first and second tests these variables were investigated during a 13 min recovery exercise at 60% and 40% of the preceding maximum rowing speed respectively. In the third test the subjects had a resting recovery. The results include a significant increase (P less than 0.001) in the rate of lactate removal following the 40% recovery compared with the 60% and resting recoveries. The HRs were significantly lower during the last minute of resting recovery compared with 40% and 60% recoveries (P less than 0.001). The same was true when 40% recovery was compared with 60% recovery (P less than 0.001). The present data suggest firstly that 40% of the maximum rowing speed is an appropriate pace for effective LA removal and secondly that, at least for trained rowers, 86% of their maximum HR can be taken as an indication of work of an intensity at or above anaerobic threshold.

Adult↗