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Synovial fluid lactic acid in septic arthritis.

Lactic acid concentrations in the synovial fluid of 71 patients with inflammatory arthritis were determined by an enzyme method. In 63 samples from 54 patients with a variety of non-septic arthritides, including rheumatoid arthritis, reactive arthritis and gout, the concentration of lactic acid was never greater than 10.2 mmol/l, whereas all twelve patients with septic arthritis had concentrations of 11 mmol/l or greater. Two patients with gonococcal arthritis did not have raised lactic acid concentrations. The enzyme method of lactic acid estimation is an accurate reproducible means of differentiating septic from nonseptic arthritis prior to the isolation of the infecting organism. However, caution is necessary when interpreting the results in those patients who have recently received antibiotic therapy, or in whom gonococcal arthritis is suspected.

Arthritis, Infectious↗

rDNA ITS sequence of Rhizopus oryzae: its application to classification and identification of lactic acid producers.

Rhizopus oryzae is an important organism for its production of organic acids such as lactic acid, fumaric acid, etc. To date, there were no easy methods to classify strains according to their acid production. The sequences of the ribosomal RNA-encoding DNA (rDNA) internal transcribed spacer (ITS) region of 64 strains of R. oryzae were analyzed and found to conserve mutations correspond to acid production. We have devised a way to use these mutations for a novel method to identify lactic-acid-producing Rhizopus oryzae, by designing specific polymerase chain reaction (PCR) primers on them. Touch down PCR using these primers amplified the ITS DNA of lactic acid producers specifically. By this method, we could isolate lactic acid producing strains from Indonesian fermented foods.

Base Sequence↗

Part III. Direct enzymatic esterification of lactic acid with fatty acids.

Lipase catalyzed esterification reactions between lactic acid and several fatty acids have been studied. Difficulties arise in esterifying lactic acid because of the potential for this substance to act both as an acyl donor and as a nucleophile. These difficulties were minimized via strategies which greatly increased the yield of the desired ester. Use of the companion fatty acid in excess with respect to lactic acid in an apolar solvent (n-hexane) in which the lactic is not completely dissolved has been employed to minimize the potential for lactic acid to act as an acyl donor in a self-polymerization reaction.Beneficial and sinergistic effects of both silica gel and molecular sieves on conversion to the desired product are described. However, careful control of the amount of molecular sieves used is required. This fact is a consequence of two opposing effects of this material: i.e. adsorption of both lactic acid and water from the reaction mixture. For reaction between caprylic and lactic acids, use of an excessive amount of enzyme reduces the extent of conversion to 2-O-caproyl-lactic acid.A very pure ester of the L-enantiomer (optical rotation of [alpha]D(25) = -23.5) can be prepared in n-hexane using a four fold excess of caprylic acid and Candida antarctica lipase. Optimum reaction conditions lead to 35% yield of 2-O-caproyl-lactic acid, a result which is close to the maximum yield that can be enantioselectively obtained from commercial grade lactic acid (68 mole per cent monomer).

Journal Article↗

Organization and regulation of genes for amino acid biosynthesis in lactic acid bacteria.

The recent description of large clusters of biosynthetic genes in the chromosome of Lactococcus lactis and, to a lesser extent, of Lactobacillus, has brought some information on gene organization and control of gene expression in these organisms. The genes involved in a given amino acid biosynthetic pathway are clustered at a single chromosomal location and form an operon. Additional genes which are not required for the biosynthesis are present within some operons. Genetic signals are, in general, similar to those found in other prokaryotes. Several systems controlling gene expression have been identified and transcription attenuation seems frequent. Among the attenuation mechanisms identified, one resembles that controlling amino acid biosynthesis in many bacteria by ribosome stalling at codons corresponding to limiting amino acid. The others are different and might be related to a new class of attenuation mechanism. Preliminary evidence for a new type of regulatory mechanism, involving a metabolic shunt, is also reviewed.

Amino Acids↗

Novel method of lactic Acid production by electrodialysis fermentation.

In lactic acid fermentation by Lactobacillus delbrueckii, the produced lactic acid affected the lactic acid productivity. Therefore, for the purpose of alleviating this inhibitory effect, an electrodialysis fermentation method which can continuously remove produced lactic acid from the fermentation broth was applied to this fermentation process. As a result, the continuation of fermentation activity was obtained, and the productivity was three times higher than in non-pH-controlled fermentation. In electrodialysis fermentation, the amount of produced lactic acid was 82.2 g/liter, which was about 5.5 times greater than that produced in non-pH-controlled fermentation. It was concluded that these good results were obtained on account of alleviating the lactic acid inhibitory effect by electrodialysis fermentation. However, the fouling of anion-exchange membranes by cells was observed in electrodialysis fermentation.

Journal Article↗

Morphology and biodegradability of a binary blend of poly((R)-3-hydroxybutyric acid) and poly((R,S)-lactic acid).

The miscibility, morphology, and biodegradability of a binary blend of bacterial poly((R)-3-hydroxybutyric acid)(P((R)-3HB);Mn = 300,000) with atactic poly((R,S)-lactic acid)(P((R,S)-LA);Mn = 9,000) were studied by means of differential scanning calorimetry, optical microscopy, scanning electron microscopy, and hydrolysis with an without enzyme. Differential scanning calorimetry revealed that a P((R)-3HB)-P((R,S)-LA) blend had a single glass-transition temperature for all proportions of the components. The spherulites of P((R)-3HB) were volume filled in the blend films, indicating the inclusion of amorphous P((R,S)-LA) within the spherulites. The spherulitic growth rate decreased with an increase in the content of P((R,S)-LA). These results indicate that the P((R)-3HB)-P((R,S)-LA) blend is miscible in the melt and in the amorphous state. The enzymatic hydrolysis of P((R)-3HB)-P((R,S)-LA) blend films was carried out at 37 degrees C for 19 h in 0.1 M potassium phosphate buffer (pH 7.4) with an extracellular poly(hydroxybutyrate) depolymerase from Alcaligenes faecalis T1. The rate of enzymatic surface erosion decreased with increasing P((R,S)-LA) content in the blend films. The simple hydrolysis of P((R)-3HB)-P((R,S)-LA) blend films without enzyme was also conducted at 37 degrees C in a 0.01 M potassium phosphate buffer (pH 7.4) for 150 days. The hydrolytic scission of P((R)-3HB) polymer chains was accelerated by blending with P((R,S)-LA). However, the rate of enzymatic hydrolysis was much faster than the rate of nonenzymatic hydrolysis.

Alcaligenes↗

Antimicrobial interactions of microbial species involved in the fermentation of cassava dough into agbelima with particular reference to the inhibitory effect of lactic acid bacteria on enteric pathogens.

Lactic acid bacteria, Bacillus species and yeasts are involved in the fermentation of cassava dough into agbelima. Microbial interactions within and between these groups of microorganisms were investigated in addition to the survival of five enteric pathogens inoculated into agbelima under various conditions. Nine out of 10 cultures of lactic acid bacteria isolated at the end of agbelima fermentation showed inhibitory effect against 10 cultures of lactic acid bacteria isolated at the start of fermentation. Only 3 out of 10 isolates of Bacillus subtilis were inhibited by 10 isolates of lactic acid bacteria tested. No interactions were observed between yeasts and the lactic acid bacteria, whereas three of the Bacillus isolates showed inhibitory effects against the yeasts. Twelve isolates of Lactobacillus plantarum tested inhibited the growth of an isolate each of Lactobacillus fermentum and Lactobacillus brevis but none tested positive for bacteriocin production. The antimicrobial effect of the lactic acid bacteria was attributed to acid production. In fermenting cassava dough, enteric pathogens survived to different extents depending on pH and their sensitivity to acids. Vibrio cholerae C-230, Salmonella typhimurium 9 and Salmonella enteritidis 226 were not detectable in 10 g of sample after 4 h when inoculated into the 48-h fermented product, agbelima, whereas Shigella dysenteriae 2357T and Escherichia coli D2188 were detectable up to 8 h in the product.

Antibiosis↗

Apparent antifungal activity of several lactic acid bacteria against Penicillium discolor is due to acetic acid in the medium.

Fifty-six dairy bacteria belonging to the genera Lactococcus, Lactobacillus, Pediococcus, Propionibacterium, Streptococcus, Enterococcus, Leuconostoc, and Brevibacterium were screened for antifungal activity against four species of fungi relevant to the cheese industry (Penicillium discolor, Penicillium commune, Penicillium roqueforti, and Aspergillus vesicolor). Most of the active strains belonged to the genus Lactobacillus, whereas Penicillium discolor was found to be the most sensitive of the four fungi investigated. Further studies on P. discolor showed antifungal activity only below pH 5. This effect of pH suggests that organic acids present in the culture could be involved in the detected activity. Determination of acid composition revealed lactic acid production for active dairy strains and the presence of acetic acid in active as well as inactive strains. It was demonstrated that the undissociated acetic acid originates from the bacterial growth medium. The synergistic effect of the acetic acid present and the lactic acid produced was likely the main factor responsible for the antifungal properties of the selected bacteria. These results could explain some discrepancies in reports of the antifungal properties of lactic acid bacteria, since the role of acetic acid has not been considered in previous studies.

Acetic Acid↗

Shear-load carrying capacity of cancellous bone after implantation of self-reinforced polyglycolic acid and poly-L-lactic acid pins: experimental study on rats.

Both distal femora of 40 rats were implanted with a self-reinforced polyglycolic acid (SR-PGA) pin in the right femur and with a self-reinforced poly-L-lactic acid (SR-PLLA) pin in the left femur. The intact femora of 20 rats served as controls. The follow-up times were 1, 3, 6, 12, 24, 36, 48, and 52 weeks. After killing all operated and control femora were examined macroscopically and radiographically. The shear-load carrying capacities of all the femurs were investigated, and the SR-PGA and SR-PLLA fixed specimens were compared with each other and with the control specimens. The shear-load carrying capacities reached their highest values at 36 weeks in the SR-PGA and SR-PLLA fixed and control specimens. Thereafter they gradually decreased. At 52 weeks both the SR-PGA fixed specimens and the control specimens had statistically significantly (p < 0.001) higher values than the SR-PLLA fixed specimens, when the influence of the pins had ceased. Otherwise, the shear-load capacities showed higher values in the SR-PLLA fixed specimens, as the pins carried the load. During the whole follow-up period the mean shear-load carrying capacity of the SR-PGA fixed specimens was 171.2 N and that of the SR-PLLA fixed specimens 180.9 N, the corresponding value of the control specimens was 148.2 N.

Animals↗

Effects of in vivo and in vitro production of lactic acid on ionized, protein-bound, and complex-bound calcium in blood.

We have studied, both in vitro and in vivo, the quantitative effects of lactic acid production on concentrations of ionized calcium, bound calcium, pH, bicarbonate, and albumin. To do so, we examined the effects of addition of aqueous solutions of either hydrochloric acid, lactic acid, or lithium lactate to blood; we studied in vitro accumulation by storing blood sealed in tubes at room temperature for 5 h, then exposing the blood to air; and we induced in vivo production of lactic acid in healthy individuals who climbed stairs for 10 min. Lactic acid evidently affects the ionized, protein-bound, and complex-bound calcium concentrations in the following ways: (a) hydrogen ions from lactic acid bind to protein, which decreases protein-bound calcium; (b) lactate chelates calcium ions from free ionized calcium and protein-bound calcium about equally; and (c) the loss of a millimole of bicarbonate, either by exposure of blood to air or by respiratory alkalosis, results in the release of about 7 mumol of calcium ions, which re-equilibrate with both the protein-bound and ionized calcium. Because lactate apparently removes calcium ions directly from albumin, our study indicates that protein-bound calcium readily provides calcium ions that buffer changes in the concentration of ionized calcium.

Acidosis, Lactic↗

Roles of silica gel in polycondensation of lactic acid in organic solvent.

Poly(lactic acid) is among the most important biodegradable, biocompatible polymers. To explore the feasibility of making poly(lactic acid) through potentially more selective enzymatic methods, the lipase-catalyzed direct polycondensation of lactic acid in organic solvents was investigated. At 37 degrees C the reaction was found to favor nonpolar solvents with larger log P values and smaller log S(w/o values. The addition of silica gel appeared to greatly enhance the lactic acid conversion (up to 98%) and the lipase stability under the reaction condition. However, upon further investigations, the silica gel itself was found to catalyze the polycondensation, in addition to the role of water removal. The conversion catalyzed by silica gel alone was actually higher than that by silica gel + lipase (or lipase alone). Up to 93% conversion of the acid functional group (or about 99.5% conversion of lactic acid monomer) was obtained in 120 h with silica gel as the catalyst. The finding is especially significant for interpreting (or reconsidering) the results of many presumably enzyme-catalyzed organic-phase reactions in the presence of silica gel.

Biocompatible Materials↗