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Respiratory effects of lactic acid injected into the jugular vein of newborn rabbits.

To define respiratory effects of peripheral acidemia during early development, we injected lactic acid (LA) into the jugular vein of 32 1- to 7-d-old rabbit pups and four adult rabbits. The body weight of newborns varied between 67 +/- 17 g in 1-d-old and 119.8 +/- 33.7 g in 7-d-old animals versus 4 +/- 0.1 kg in adults. Animals were anesthetized with ketamine (30 mg/kg) and urethane (0.6-1.2 g/kg), tracheotomized, and breathed spontaneously. Diaphragmatic electromyogram, tidal volume, esophageal pressure, and arterial pressure were recorded. In adult rabbits, a LA dose of 0.25 mM/kg elicited an increase in the rate of breathing with no change or a decrease in tidal volume (early response), followed by a deep and fast respiration (late response). The same LA dose had no effect or provoked only the late response in 1- and 2-d-old rabbits and led to the early and late responses in older newborns. The early response was abolished and the late response was diminished after vagotomy. Our results suggest that the early response is mediated by vagal afferents and the late response, although modified by vagal input, is of an extravagal origin. It is suggested that acidemia may be involved in the transient tachypnea syndrome in newborn infants. Possible contribution of vagal nonmyelinated endings in triggering the early response to LA is discussed.

Acidosis↗

Comparative studies of class IIa bacteriocins of lactic acid bacteria.

Four class IIa bacteriocins (pediocin PA-1, enterocin A, sakacin P, and curvacin A) were purified to homogeneity and tested for activity toward a variety of indicator strains. Pediocin PA-1 and enterocin A inhibited more strains and had generally lower MICs than sakacin P and curvacin A. The antagonistic activity of pediocin-PA1 and enterocin A was much more sensitive to reduction of disulfide bonds than the antagonistic activity of sakacin P and curvacin A, suggesting that an extra disulfide bond that is present in the former two may contribute to their high levels of activity. The food pathogen Listeria monocytogenes was among the most sensitive indicator strains for all four bacteriocins. Enterocin A was most effective in inhibiting Listeria, having MICs in the range of 0.1 to 1 ng/ml. Sakacin P had the interesting property of being very active toward Listeria but not having concomitant high levels of activity toward lactic acid bacteria. Strains producing class IIa bacteriocins displayed various degrees of resistance toward noncognate class IIa bacteriocins; for the sakacin P producer, it was shown that this resistance is correlated with the expression of immunity genes. It is hypothesized that variation in the presence and/or expression of such immunity genes accounts in part for the remarkably large variation in bacteriocin sensitivity displayed by lactic acid bacteria.

Amino Acid Sequence↗

Enzymatic, alkaline, and autocatalytic degradation of poly(L-lactic acid): effects of biaxial orientation.

The hydrolytic degradation of biaxially oriented and de-oriented (melt-crystallized) poly(l-lactic acid) (PLLA) films was investigated in Tris-HCl-buffered solution (pH 8.6) with proteinase K, alkaline solution, and phosphate-buffered solution (pH 7.4) by the use of gravimetry, gel permeation chromatography, differential scanning calorimetry, and scanning electron microscopy. Biaxial orientation disturbed the proteinase K-catalyzed enzymatic degradation of PLLA films and the effects of biaxial orientation overcame those of crystallinity. The former may be due to the fact the enzyme cannot attach to the extended (strained) chains in the amorphous regions of the biaxially oriented PLLA film or cannot catalyze the cleavage of the strained chains. Another probable cause is that the enzyme can act only at the film surface of the biaxially oriented PLLA film, in marked contrast with the case of the de-oriented PLLA films where enzymatic degradation can proceed beneath the spherulitic crystalline residues. The effects of biaxial orientation on the alkaline and autocatalytic degradation of the PLLA films were insignificant for the periods studied here. The crystallinity rather than the biaxial orientation seems to determine the alkaline and autocatalytic degradation rates of the PLLA films. The accumulation of crystalline residues formed as a result of selective cleavage and removal of the amorphous chains was observed for the de-oriented PLLA films, but not for the biaxially oriented PLLA film, when degraded in the presence of proteinase K. This means the facile release of formed crystalline residues from the surface of the biaxially oriented PLLA film during enzymatic degradation, due to the fact that the crystalline regions of the biaxially oriented PLLA film were oriented with their c axis parallel to the film surface.

Alkalies↗

Mutagenesis in the post genomics era: tools for generating insertional mutations in the lactic acid bacteria.

The increasing availability of whole genome sequences has increased the demand for effective tools to generate insertional mutations in the lactic acid bacteria (LAB). Several novel approaches, such as shuttle-, transposome- and intron-based mutagenesis methods, are possible additions to the existing repertoire of transposon- and recombination-based tools available for mutagenesis of LAB.

Bacteria↗

[Determination of serum lactic acid and the acid-base equilibrium in carbon monoxide poisoning].

The authors present a description of severe accidental carbon monoxide poisoning. They stress the significance of the examination of lactic acid level at this poisoning. In case described the level of carbonyl haemoglobin could not be determined because of device trouble. Patient AA (16-year-old) was admitted in relatively very good condition, without clinical signs of severer affection. Considering the facts that gas-poisoning (over 24 hrs) was found through anamnesis, and simultaneous admission of patient's greatgrandmother in critical condition, the patient AA was treated also with oxygen of high concentration, monitored, kept in absolute rest. High lactate level (18.9 mmol/l) and the evidence of metabolic acidosis (pH 7.16, pO2 8.65, pCO2 2.06, sBE -22.1) made to suspect of severe poisoning. Despite intensive oxygenotherapy it came to severe left-side cardial failure associated with pulmonal edema, anuria, shock after 4 hours, managed successfully within 4 hours. Lactate level correlated very well with condition severity. Septum hypokinesis was demonstrated cardiografically and it disappeared within 6 days. However, symetric inversion of T wave in many leads (I, II, aVF, V 2-6) persisted for 24 days. The authors assume that routine examination of acid base balance and lactate level in addition to the determination of carbonyl haemoglobin level is very important for the consideration of the severity of carbon monoxide poisoning. Lactataemia determination is probably even more significant for the assumption of oxygen deficit and therefore for the total prognosis of the patient.

Accidents, Home↗

Porous poly (DL-lactic acid) modified chitosan-gelatin scaffolds for tissue engineering.

Chitosan-gelatin (Cs-Gel) scaffolds are modified with poly (DL-lactic acid) (PDLLA) dichloromethane solution of different concentrations (0.1, 0.5, and 1.0%) and immersed in water after the evaporation of the solvent. The swollen scaffolds are freeze dried. The concentration of PDLLA has significant effects on both the physicomechanical properties and the cytocompatibility. Data reveal that only the 0.1% concentration could increase the tensile strength fourfold in comparison with the pristine Cs-Gel scaffold, while maintaining the human fibroblast adhesion, migration, and proliferation just like the Cs-Gel scaffold.

Absorption↗

Lactic acid triggers, in vitro, thiomersal to degrade protein in the presence of PLGA microspheres.

Microspheres of polymers like poly(lactic-co-glycolic acid) (PLGA) have been studied as a vehicle for controlled release vaccines. They require materials and processes that might change the protein antigenicity. Lactic acid is produced during microsphere degradation that occurs in tandem with protein liberation. In addition, most of the proteins that have been used in microencapsulation studies contain Thimerosal((R))(TM) and this can introduce another undesirable effect for their stability. We demonstrated in vitro that the thiosalycilic acid (TSA), produced after the reduction of TM by lactic acid, reduces the S-S bridge of the previously incubated diphtheric toxoid (Dtxd). This reduction is immediately followed by blocking the two -SH formed by the same TSA molecules. In the light of these conclusions it is necessary now, to reinterpret the in vitro protein degradation-stabilization data in the presence of PLGA microspheres, mainly for those proteins which contain S-S. We propose that all the PLGA microspheres microencapsulation studies and protein structural considerations should be done in the absence of TM as preservative.

Diphtheria Toxoid↗

Biocatalysis using an organic-soluble enzyme for the preparation of poly(lactic acid) in organic solvents.

Proleather from Bacillus sp. was chemically modified with decanoyl chloride for enhanced activity for the preparation of poly(lactic acid) in organic solvents. The modified enzyme was highly soluble (up to 44 mg-protein/ml) and active in various organic solvents including chloroform, tetrahydrofuran (THF), pyridine and acetone. The organic-soluble proleather efficiently catalyzed the polymerization of ethyl lactate. The reaction rate was 4-22 times that of native proleather, depending upon the solvent applied. The solubilized enzyme showed a highest activity at 50 degrees C, the same optimum temperature for both the native proleather and an immobilized enzyme, Novozyme-435. Denaturation of the enzymes' protein structures appeared to be the critical factor regulating the optimum activity temperature. Differential scanning calorimetry (DSC) analyses of the enzymes showed endothermic peaks around 55 degrees C, indicating the proteins' structures altered in that temperature range. Interestingly, the activity of the solubilized enzyme showed a more complicated water dependence as compared to native proleather.

Acetone↗

Ciprofloxacin-impregnated poly-L-lactic acid drug carrier. New aspects of a resorbable drug delivery system in local antimicrobial treatment of bone infections.

Resorbable poly-L-lactic acid (PLLA) cylinders (3.5 mm diameter, 5 mm in length) carrying 6% of weight ciprofloxacin (Ciprobay, Bayer AG, Leverkusen, FRG) were investigated in vitro to explore their properties as a slow-release antibiotic deposit. Forty bioactive cylinders stored in test tubes were covered with phosphate buffer (pH 7.4 at 37 degrees C) and 40 with fresh human blood plasma and tested under various conditions. For comparison a gentamicin-polymethylmethacrylate (PMMA) chain (Septopal, E. Merck, Darmstadt, FRG) was exposed to similar test conditions. The quantities of ciprofloxacin and gentamicin released were analysed by a microbiological method (bioassay). The concentrations of ciprofloxacin released were analysed by a microbiological method (bioassay). The concentrations of ciprofloxacin released from 40 cylinder were initially very high (up to 180 mg/l) but they decreased rapidly within the first 5 days (4.2-22.5 mg/l). Early release of gentamicin reached up to 227.5 mg/l but dropped to of 22 mg/l on the 14th day. Complete degradation of the PLLA-cylinders was not seen in the observed period of 92 days. The mean loss of mass was 8.4%. The recovery of incorporated ciprofloxacin was 6.5% on average.

Biological Assay↗

Spray-drying of bacteriocin-producing lactic acid bacteria.

Cell survival, cellular damage, and antagonistic activity were investigated after spray-drying of four bacteriocin-producing strains of lactic acid bacteria: Lactococcus lactis subsp. lactis 140, isolated from natural whey culture and producing a narrow-inhibitory spectrum bacteriocin); L. lactis subsp. lactis G35, isolated from pizza dough and producing nisin; Lactobacillus curvatus 32Y and Lactobacillus sp. 8Z, isolated from dry sausages. Trials were performed with bacteria suspended in skimmed milk or directly grown in whey. Three air temperatures at the inlet of the drier (160, 180, and 200 degrees C) and three flow rates (10, 13, and 17 ml/min) were assayed. Cell viability and bacteriocin activity of the dried materials were determined immediately after the process and after 5, 15, 30, and 60 days of storage at 4 degrees C. There was no significant difference between the two feeding suspensions in cell survival, always decreasing with the increase of inlet-air temperature. No loss of bacteriocin activity was detected in reconstituted powders, nor was any loss of ability to produce bacteriocin found after drying. Investigations of sensitivity to NaCl revealed only temporary damage to dried bacteria. During storage for 2 months at 4 degrees C, all samples, but mainly the lactococcal strains, displayed a gradual decrease in cell survival. Bacteriocin activity remained at the same level, allowing powders to be considered as effective biopreservatives.

Aerosols↗

Effects of particle size, helium gas pressure and microparticle dose on the plasma concentration of indomethacin after bombardment of indomethacin-loaded poly-L-lactic acid microspheres using a Helios gun system.

We investigated the effects of the particle size of indomethacin-loaded poly-L-lactic acid microspheres (IDM-loaded PLA MS), the helium pressure used to accelerate the particles, and the bombardment dose of PLA MS on the plasma concentration of IDM after bombarding with IDM-loaded PLA MS of different particle size ranges, 20-38, 44-53 and 75-100 microm, the abdomen of hairless rats using the Helios gene gun system (Helios gun system). Using larger particles and a higher helium pressure, produced an increase in the plasma IDM concentration and the area under the plasma concentration-time curve (AUC) and resultant F (relative bioavailability with respect to intracutaneous injection) of IDM increased by an amount depending on the particle size and helium pressure. Although a reduction in the bombardment dose led to a decrease in C(max) and AUC, F increased on decreasing the bombardment dose. In addition, a more efficient F was obtained after bombarding with IDM-loaded PLA MS of 75-100 microm in diameter at each low dose in different sites of the abdomen compared with that after bolus bombardment with a high dose (dose equivalent). These results suggest that the bombardment injection of drug-loaded microspheres by the Helios gun system is a very useful tool for delivering a variety of drugs in powder form into the skin and systemic circulation.

Air Pressure↗

Effect of polymer/basic drug interactions on the two-stage diffusion-controlled release from a poly(L-lactic acid) matrix.

We investigated the effect of drug physico-chemical properties on the release of basic drugs from poly(L-lactic acid) (P(L)LA) cylindrical matrices (rods; 10 mmx1 mm diameter). All the rods were revealed to exhibit two-stage diffusion-controlled release profiles resulting from the transformation of P(L)LA from an amorphous to a semicrystalline state in aqueous medium. On the assumption that interactions between polymer carboxyl residues and basic drugs control the drug release rate, we evaluated the strength of these interactions by the drug partition between the polymer and the aqueous medium. In the first release stage, the drugs diffused through the swollen polymer matrix. The polymer-drug interactions shielded the polymer terminal carboxyl residues, thereby resulting in a less hydrated matrix and consequent diminishment of drug diffusion. In the second release stage, the drugs diffused through the water-filled micropores which had developed as a result of polymer crystallization. The stronger polymer-basic drug interactions reduced the drug diffusion rate by decreasing not only the porosity of the matrix, but also the drug partition to the water-filled micropores. It was also found that the fractional drug release rate in the second stage increased with drug content of the rod at the pH where both the polymer carboxyl residues and the drugs were ionized. Since the polymer-drug interactions must be close to saturation with increasing drug content, we believe this result to be due to an increase in the ratio of the drug partition to the water-filled micropores.

Biocompatible Materials↗

Ability of dairy strains of lactic acid bacteria to bind a common food carcinogen, aflatoxin B1.

This study was conducted to examine the ability of selected dairy strains of lactic acid bacteria to remove aflatoxin B1 (AFB1) from liquid media. Both Lactobacillus rhamnosus strain GG (LBGG) and L. rhamnosus strain LC-705 (LC705) can significantly (P > 0.05) remove AFB1 when compared with that by other strains of either Gram-positive or Gram-negative bacteria. Removal of AFB1 by LBGG and LC705 was a rapid process with approximately 80% AFB1 removed at 0 hr. Removal of AFB1 by these two strains was both temperature and bacterial concentration dependent.

Aflatoxin B1↗

Fibronectin immobilization using water-soluble carbodiimide on poly-L-lactic acid for enhancing initial fibroblast attachment.

The aim of this study is to evaluate the influence of fibronectin immobilization on poly-L-lactic acid (PLA) films on the initial attachment of human gingival fibroblasts. Carboxylic acid groups are chemically introduced on the PLA films' surface by surface hydrolysis with 0.5 M NaOH. The contact angle of PLA surface with respect to double-distilled water decreases significantly after NaOH hydrolysis. X-ray photoelectron spectroscopy (XPS) also reveals significantly higher intensities of C(C=O)/C(C-O) after NaOH hydrolysis. Fibronectin is immobilized onto the hydrolyzed PLA surface through a condensation reaction between the carboxylic acid groups on the hydrolyzed PLA surface and the amino groups of fibronectin using water-soluble carbodiimide. XPS analysis shows that the fibronectin-immobilized PLA surface is enriched with nitrogen atoms. The immobilization of fibronectin significantly enhances the number of initially attached human gingival fibroblasts on the PLA surface. No obvious differences in morphology are noted between fibroblasts cultured on native PLA and on fibronectin-immobilized PLA. Fibronectin can be immobilized onto the PLA surface after NaOH hydrolysis and this is effective in enhancing the initial attachment of human gingival fibroblasts.

Adsorption↗

A preliminary study of the effect of glass-ionomer and related dental cements on the pH of lactic acid storage solutions.

Glass-ionomer cements, both self-hardening and resin-modified, have been shown to increase the pH of lactic acid solutions in which they are stored. Similar results have been obtained for a zinc phosphate and a zinc polycarboxylate cement. The pH was increased over a period of 7 days by between 1.54 and 2.65 pH units from an initial value of pH of 2.60, depending on the cement. It is concluded that, as a result of this ability to neutralize surrounding aqueous solutions, these materials may have the beneficial effect in vivo of inhibiting caries development. In the case of glass-ionomers, this mechanism might complement that of fluoride release.

Dental Caries↗

Isolation and identification of lactic acid bacteria with effect of immune protection to Eschericia coli in mice.

Lactic acid bacteria were isolated from an alcohol fermentation broth, and the activity as a probiotic was examined using pathogenic E. coli. Thirty-six strains exhibiting good growth were isolated in the medium of concentrated mush which was a residue resulted in the alcohol distillation process. One of these strains, Lactobacillus paracasei subsp. paracasei I-5, could be grown in the medium containing 8 vol% ethanol and at 45 degrees C. The characteristics were different from the type strain, L. paracasei subsp. paracasei NBRC 15889. L. paracasei I-5 showed an excellent growth in the concentrated mush, which just diluted two-fold and adjusted the pH. ICR mice were fed with a standard germ-free feed (CMF) and the strain I-5 (7 x 10(9) cells/day) was orally administrated for 11 days prior to the intraperitoneal challenge with pathogenic E. coli Juhl. After the challenge, mice administrated the strain I-5 exhibited a high survival rate and survival extension days (p < 0.01) compared with the control. The results suggested that the strain might enhance the animal resistance against microbial pathogens. Neonatal diarrhea caused by E. coli is a serious disease in calf breeding. The strain might be practically valuable to prevent diarrhea in calves.

Anaerobiosis↗

A simple test for urinary lactic acid.

A new simple rapid test for the detection of increased urinary lactic is described. The method depends on the transformation of lactic acid to acetaldehyde by cesium IV. Acetaldehyde reacts with sodium nitroprusside prducing a blue color. Patients from an emergency ward and patients with experimentally induced hyperlactatemia have been screened with this test. The results confirm that there exists a correlation between blood and urinary lactate levels and that a clinically relevant hyperlactemia is associated with an increased urinary lactate concentration which can be reliably detected with this test.

Acidosis↗

Ester synthesis in an aqueous environment by Streptococcus thermophilus and other dairy lactic acid bacteria.

The ability of Streptococcus thermophilus ST1 and 19 other dairy lactic acid bacteria (LAB) to synthesize esters was investigated in an aqueous environment. These LAB were able to synthesize esters from alcohols and glycerides via a transferase reaction (alcoholysis) in which fatty acyl groups from glycerides were transferred to alcohols. S. thermophilus ST1 was active on tributyrin and on di- or monoglycerides of up to C10 with ethanol as the acyl acceptor. This strain was also active on a diglyceride of C6 and monoglyceride of C8 with 2-phenyl ethanol as the acyl acceptor. Alcoholysis occurred preferentially over hydrolysis. S. thermophilus ST1 had an apparent K(m) value of 250 mM for ethanol and an apparent K(m) value of 1.3 mM for tributyrin, measured against whole cells. Around 80% of both the transferase activity and the esterase activity were detected in the cell-free extract (CFE) of strain ST1. Both activities in the CFEs of five LAB tested were, to a similar degree, enhanced slightly by growth in the presence of ethanol and tributyrin. Using tributyrin and ethanol as substrates, the transferase activities ranged over 0.006-1.37 units/mg cell dry weight among the LAB tested and were both species- and strain-dependent.

Alcohols↗