Optimization of exopolysaccharide production by Lactobacillus delbrueckii subsp. bulgaricus RR grown in a semidefined medium

The role of K+ channels in the nitric oxide-independent renal vasodilator effect of acetylcholine (Ach) was examined to address the hypothesis that the mechanism underlying this response was different from that of bradykinin, because an earlier study indicated the possibility of different mediators. We used the rat isolated, perfused kidney that was constricted with phenylephrine and treated with nitroarginine and indomethacin to inhibit nitric oxide synthase and cyclooxygenase, respectively. The nonspecific K+ channel inhibitors, procaine and tetraethylammonium (TEA), reduced vasodilator responses to Ach and cromakalim, but not those to nitroprusside. Glibenclamide, an inhibitor of ATP-sensitive K+ channels, reduced vasodilator responses to cromakalim but did not affect those to Ach or nitroprusside. Charybdotoxin, an inhibitor of Ca(++)-activated K+ channels, reduced vasodilator responses to Ach without affecting those to cromakalim or nitroprusside. Iberiotoxin and apamin, inhibitors of large- and small-conductance Ca(++)-activated K+ channels, respectively, did not reduce vasodilation induced by Ach, cromakalim or nitroprusside. The inhibitor of cytochrome P450, clotrimazole, reduced the renal vasodilator effects of Ach and bradykinin but not those of nitroprusside or SCA 40, an agonist for Ca(++)-activated K+ channels. These results suggest that in the rat kidney, Ach, like bradykinin, utilizes a charybdotoxin-sensitive Ca(++)-activated K+ channel of intermediate conductance to elicit vasodilation and that this effect may be dependent on cytochrome P450 activity.     

The prolyl aminopeptidase from Lactobacillus delbrueckii subsp. bulgaricus belongs to the alpha/beta hydrolase fold family

Prolyl aminopeptidase (PepIP) of Lactobacillus delbrueckii subsp. bulgaricus displays the Gly-x-Ser-x-Gly-Gly consensus motif surrounding the catalytic serine of the prolyl oligopeptidases family. Sequence comparison revealed that this motif and two other domains appear well conserved among bacterial PepIPs and members of the alpha/beta hydrolase fold family. Secondary structural predictions of PepIP were performed from amino acid sequence and corroborated by circular dichroism analysis. These predictions well matched the core structure of alpha/beta hydrolases organised in eight beta-sheets connected by alpha-helices. We obtained 26 mutants of PepIP by chemical or site-directed mutagenesis. Most substitutions associated with stable and inactive mutant proteins were mainly located in the three conserved boxes (including the catalytic serine motif). Taken together, our results strongly suggest that PepIP belongs to the alpha/beta hydrolase fold family and that Ser107, Asp246 and His273 constitute the catalytic triad of the enzyme.     

Note: identification of Lactobacillus delbrueckii subspecies bulgaricus and subspecies lactis dairy isolates by amplified rDNA restriction analysis

Amplification and restriction analysis of the 16S rRNA gene (PCR-ARDRA) was used to identify Lactobacillus delbrueckii dairy isolates and to assign them to its three subspecies (delbrueckii, lactis and bulgaricus). PCR-ARDRA allowed confirmation of predictions from the DNA database regarding the restriction mapping of the 16S sequence to be verified in practice. PCR-ARDRA was shown to be a reliable and rapid method for identifying Lact. delbrueckii isolates at the subspecies level and for differentiating this species from Lact. helveticus and Lact. acidophilus.     

Characterization of a prolate-headed bacteriophage of Lactobacillus delbrueckii subsp. lactis, and its DNA homology with isometric-headed phages

A new Lactobacillus delbrueckii subsp. lactis bacteriophage, JCL 1032, was characterized. JCL 1032 had a small, elongated prolate head, and a long non-contractile tail with cross-bars. The restriction map of JCL 1032 genome was constructed with five endonucleases. The genome was 45.8 kb in size, and it had cohesive ends (cos). Molecular masses of the phage structural proteins were also determined. JCL 1032 showed DNA homology with morphologically dissimilar, isometric-headed phages of Lb. delbrueckii (subsp. lactis and subsp. bulgaricus) when analyzed by Southern hybridization. Although in general JCL 1032 was only distantly related to isometric-headed phages, there were also a few short highly homologous (minimal homology 84%) DNA regions.     

Cell surface characteristics of Lactobacillus casei subsp. casei, Lactobacillus paracasei subsp. paracasei, and Lactobacillus rhamnosus strains

Hydrophilic and electrostatic cell surface properties of eight Lactobacillus strains were characterized by using the microbial adhesion to solvents method and microelectrophoresis, respectively. All strains appeared relatively hydrophilic. The strong microbial adhesion to chloroform, an acidic solvent, in comparison with microbial adhesion to hexadecane, an apolar n-alkane, demonstrated the particularity of lactobacilli to have an important electron donor and basic character and consequently their potential ability to generate Lewis acid-base interactions with a support. Regardless of their electrophoretic mobility (EM), strains were in general slightly negatively charged at alkaline pH. A pH-dependent behavior concerning cell surface charges was observed. The EM decreased progressively with more acidic pHs for the L. casei subsp. casei and L. paracasei subsp. paracasei strains until the isoelectric point (IEP), i.e., the pH value for which the EM is zero. On the other hand, the EM for the L. rhamnosus strains was stable from pH 8 to pH 3 to 4, at which point there was a shift near the IEP. Both L. casei subsp. casei and L. paracasei subsp. paracasei strains were characterized by an IEP of around 4, whereas L. rhamnosus strains possessed a markedly lower IEP of 2. The present study showed that the cell surface physicochemical properties of lactobacilli seem to be, at least in part and under certain experimental conditions, particular to the bacterial species. Such differences detected between species are likely to be accompanied by some particular changes in cell wall chemical composition.     

Characterization of pepR1, a gene coding for a potential transcriptional regulator of Lactobacillus delbrueckii subsp. lactis DSM7290

This article examines the importance of economic factors in physician and other health service utilization among older adults living in Bermuda. Using data drawn from a national survey of 500 Bermudian elders, the findings reveal economic factors to be unrelated to the use of physician services directly, but to be significant determinants of the use of other health services. Need plus non-need factors are related to the use of both physician and other health services. The employer-based system does seem to provide universal access, but factors in addition to need influence the receipt of care.     

德氏乳杆菌特性及应用研究进展

综述了德氏乳杆菌的生物学特性,生理功能及在食品发酵、饲料工业和医疗保健等领域的应用.     

Effect of citrate on growth of Lactococcus lactis subsp. lactis in milk

The effect of citrate on the growth of Lactococcus lactis subsp. lactis var. diacetylactis in milk has been investigated. Five strains of Lactococcus lactis subsp. lactis var. diacetylactis were compared to their citrate-negative variants, which lack the plasmid coding for citrate permease. In most cases, acidification kinetics and the final bacterial concentration of pure cultures of parental and variant strains did not differ significantly. Co-cultures of parental and variant strains, however, systematically tended towards the predominance of parental strains. Citrate metabolism is responsible for this change, since the predominance of citrate-positive strains was not observed in the absence of citrate. Continuous culture in milk enabled the difference in growth rates between the parental strain Lactococcus lactis subsp. lactis var. diacetylactis CDI1 and its citrate-negative variant to be quantified by following changes in the populations of the two co-cultured strains. At 26 degrees C, the growth rate of the parental strain was 7% higher than that of its citrate-negative variant. These results show that citrate metabolism slightly stimulates the growth of lactococci in milk.     

The effects of triaxial stress on void growth and yield equations of power-hardening porous materials

In engineering applications, especially for ductile fracture of materials, nucleation, growth and coalescence of voids have often been observed. Currently there is an increase in interest for the effects of voids on the behaviour of engineering materials. In this paper, by the method of combining micro- and macro-parameters, the effects of triaxial stress on the rates of void growth and yield equations are presented for porous materials with power-hardening. The relations between triaxial stress and the rates of void growth for different n-values and yield equations with different n-values and void volume fractions are discussed. Following results have been obtained: For a porous material with power-hardening, the yield equation can be approximately expressed by an elliptical equation in equivalent stress and triaxial stress. Both the long half-axis and the short half-axis of the elliptical equation are functions of the void volume fraction for a given hardening exponent. The triaxial stress has a strong effect on the growth rates of voids. For linear hardening materials, the relation between the growth rate of voids and the triaxial stress is linear. For elastic/perfectly plastic materials with a small void volume fraction, the growth rate of voids can be described in relation to the triaxial stress with an exponential function. The results from this paper are compared with theoretical results from other researchers for elastic/perfectly plastic materials. A good agreement is shown.     

The effects of presleep stress on sleep-onset insomnia.

10 sleep-onset (SO) insomniacs and 11 noninsomniacs (18–21 yrs old) slept undisturbed for 3 nights but were exposed to brief cognitive stressors before SO on the next 2 nights. Significant between-group differences were found in responses to the cognitive stressors on self-report but not objective measures of SO latency (SOL). Noninsomniac Ss evidenced an increase and insomniac Ss a decrease in SOL on stress nights. Insomniac Ss demonstrated a significantly higher mean heart-rate response before and after the stressors. A variable expressing the relationship between objective and subjective measures of SOL (pseudoidiopathic dimension) for each S was not significantly related to Ss' responses to stress. Results are inconsistent with the hypothesized role of presleep stress in SO insomnia but are consistent with other studies indicating higher levels of physiological arousal of insomniac vs noninsomniac Ss. (22 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The effects of stress on the draw-a-person test.

External stress was hypothesized to increase manifestations of anxiety on the DAP. Secondly, it was hypothesized that there are 2 sources of these manifestations of anxiety: (a) the laboratory stress situation and (b) anxiety producing intrapsychic processes, activated by drawing the human figure. 57 male college students drew a male, female, and automobile under stress and nonstress conditions. Both hypotheses were supported; 15 and 11 of the 21 indexes significantly differentiated between stress and nonstress conditions for the male and female drawings, respectively. 5 of 17 indexes differentiated for the auto drawings. 5 indexes differentiated in the opposite direction. Suggestions were made for the use of the auto drawing in checking out clinical hypotheses in a diagnostic evaluation. (17 ref.) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Assessing the effects of stress on physical symptoms.

Stress researchers, in attempting to circumvent methodological problems in correlation studies, have resorted to analytical techniques that increase the likelihood of a Type II error. For instance, studies suggesting that stress plays no role in irritable bowel syndrome are at variance with a large body of clinical experience and may be in error. Several measurement issues are discussed, but the most significant concerns the way investigators deal with individual differences in chronic levels of stress and symptoms. J. Suls et al (see record 1994-34099-001) recommend correcting for these individual differences before estimating the relationship between stress and symptoms, but this is illogical because most stressors are chronic (i.e., they are related to the social and economic circumstances in which people live). A more important confound, not considered by most investigators, is the contribution of somatization to stress assessment and symptom reports. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Inhibitory effects of catecholamines and maternal stress on aromatase activity in the fetal rat brain

OBJECTIVES: To describe the extent of pain relief two weeks after an epidural steroid injection in patients with herniated disks and lumbar spinal stenosis, and to identify predictors of changes in pain ratings in each population. METHODS: The study design was a prospective evaluation of patients with lumbar spinal stenosis (LSS) and herniated disks (HDs) referred to a hospital-based pain clinic for an epidural steroid injection (ESI). A complete history, detailed physical examination, comprehensive pain questionnaire, and Brief Symptom Inventory were obtained for all patients. Pain was assessed at baseline and two weeks following a single ESI using a visual analog scale. RESULTS: Two hundred twelve patients (mean age 54 years) were enrolled, and 78 of these provided pain ratings before and two weeks after the injection. LSS patients improved less two weeks following the ESI than HD patients (P = 0.04). Just 38% of LSS patients reported improvement in pain score compared with 61% of HD patients. In analyses that combined LSS and HD patients, predictors of worse response included a report of health problems and a diagnosis of LSS. CONCLUSIONS: LSS patients have worse response to ESIs than HD patients. The poor response to ESI in patients with LSS underscores the need for randomized controlled trials of ESI in this population.     

Escherichia coli and Lactobacillus plantarum responses to osmotic stress

Artificial insemination using cryopreserved semen is a common management tool of the contemporary livestock producer. However, cryopreservation is detrimental to sperm function and fertility, killing some 50% of the spermatozoa during the process. Prediction of cryopreservation damage from prefreeze samples remains elusive. Computer-automated sperm head morphometry was used in this study to determine the effects of cryopreservation on bovine sperm head morphometry. Semen was collected from 18 bulls and was divided. One portion was extended to 200 x 10(6) sperm/ml and a microscope slide was prepared, while the remaining portion was cryopreserved in a Triscitrate-yolk extender. After thawing, the cryopreserved samples were prepared on microscope slides. All slides were air dried and were stained with hematoxylin and rose bengal. The morphometric dimensions for length, width, width/length, area, and perimeter for a minimum of 200 sperm heads were analyzed from each slide by computer-aided sperm head morphometry analysis, and the mean measurements were recorded. Bull sperm heads were significantly (P < 0.01) smaller in cryopreserved spermatozoa than in the companion extended samples for length (8.56+/-0.07 vs. 8.63+/-0.08 microm), width (4.39+/-0.05 vs. 4.48+/-0.05 microm), area (28.42+/-0.07 vs. 29.14+/-0.08 microm), and perimeter (23.33+/-0.21 vs. 23.70+/-0.23 microm) for all bulls. Width/length was also different (0.513 vs. 0.519). In addition, differences (P < 0.05) were found within 14 of 18 bulls for at least four of the morphometric parameters. The percent change in measures after cryopreservation were correlated (P < or = 0.05) to the variability of the extended sample. Variations in sperm head measurements were lower (P < or = 0.05) in extended samples of the four bulls in which no changes occurred than in extended samples of the remaining 14 bulls. These data suggest that the variability in sperm head measurements of individual bulls, or ejaculates, may be an indicator of sperm cryosurvivability.     

Effects of stress on morphine-elicited locomotor activity in hamsters.

Two experiments were conducted to investigate the effects of stress on morphine-elicited locomotor activity in hamsters. In Experiment 1, half of the animals were habituated to handling and injection procedures (low-stress condition) and half were not (high-stress condition) prior to 6 days of testing with a low dose (2.5 mg/kg) of morphine. On the first test day, morphine elicited hyperactivity among habituated animals, whereas among nonhabituated animals morphine elicited hypoactivity. The effects of handling diminished across test days, so that on the last test day, morphine elicited hyperactivity in both habituated and nonhabituated animals. In Experiment 2, the effect of a noise stressor on activity elicited by three doses of morphine (2.5, 5.0, and 15 mg/kg) was investigated. Half of the animals were tested under conditions of low noise stress (70 dB), and half were tested under conditions of high noise stress (90 dB). Results indicated that the effects of each dose of morphine were potentiated in the high-stress relative to the low-stress condition. Both experiments demonstrated that environmental stressors can potentiate the response to exogenous opiate administration in hamsters. Moreover, the results are consistent with the hypothesis that stress induces the release of endogenous opioids which summate with exogenous opiates to determine the final effective dose. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

In vitro evaluation of the antigenotoxic activity of a strain of L. bulgaricus isolated from commercial yogurt

Sequential oxidations at the arylamine moiety of the procainamide molecule leading to the formation of N-hydroxyprocainamide and its nitroso derivative may be responsible for lupus erythematosus observed in patients treated with the drug. The objective of the present study was to characterize major cytochrome P450 isozyme(s) involved in the N-hydroxylation of procainamide. Firstly, incubations were performed with microsomes from either lymphoblastoid cells or yeast transfected with cDNA encoding for specific human cytochrome P450 isozymes. Experiments performed with these enzyme expression systems indicated that the highest formation rate of N-hydroxyprocainamide was observed in the presence of CYP2D6 enriched microsomes. Additional experiments demonstrated that the formation rate of N-hydroxyprocainamide by CYP2D6 enriched microsomes was decreased from 45 +/- 4% to 93 +/- 1% by quinidine at concentrations ranging from 30 nM to 100 microM (all p < 0.05 vs control) and by approximately 75% by antibodies directed against CYP2D6. Secondly, incubations were performed with microsomes prepared from 15 human liver samples. Using this approach, an excellent correlation was observed between the formation rate of N-hydroxyprocainamide and dextromethorphan O-demethylase activity (CYP2D6; r = 0.9305; p < 0.0001). In contrast, no correlation could be established between N-hydroxyprocainamide formation rate and caffeine N3-demethylase (CYP1A2), coumarin 7-hydroxylase (CYP2A6), S-mephenytoin N-demethylase (CYP2B6), tolbutamide methlhydroxylase (CYP2C9), S-mephenytoin 4'-hydroxylase (CYP2C19), chlorzoxazone 6-hydroxylase (CYP2E1), dextromethorphan N-demethylase (CYP3A4), testosterone 6 beta-hydroxylase (CYP3A4/5) or lauric acid 12-hydroxylase (CYP4A11) activities. Furthermore, formation rate of N-hydroxyprocainamide was decreased in a concentration-dependent manner by quinidine (300 nM to 100 microM) and by antibodies directed against CYP2D6 but not by furafylline 20 microM (CYP1A2), ketoconazole 1 microM (CYP3A4), sulfaphenazole 10 microM (CYP2C9) or antibodies directed against CYP1A1/1A2, CYP2C, CYP2A6, CYP2E1 or CYP3A4/3A5. In conclusion, the results obtained in the present study demonstrate that CYP2D6 is the major human cytochrome P450 isozyme involved in the formation of the reactive metabolite of procainamide, namely N-hydroxyprocainamide.     

Physiological response of Lactobacillus plantarum to salt and nonelectrolyte stress

In this report, we compared the effects on the growth of Lactobacillus plantarum of raising the medium molarity by high concentrations of KCl or NaCl and iso-osmotic concentrations of nonionic compounds. Analysis of cellular extracts for organic constituents by nuclear magnetic resonance spectroscopy showed that salt-stressed cells do not contain detectable amounts of organic osmolytes, whereas sugar-stressed cells contain sugar (and some sugar-derived) compounds. The cytoplasmic concentrations of lactose and sucrose in growing cells are always similar to the concentrations in the medium. By using the activity of the glycine betaine transport system as a measure of hyperosmotic conditions, we show that, in contrast to KCl and NaCl, high concentrations of sugars (lactose or sucrose) impose only a transient osmotic stress because external and internal sugars equilibrate after some time. Analysis of lactose (and sucrose) uptake also indicates that the corresponding transport systems are neither significantly induced nor activated directly by hyperosmotic conditions. The systems operate by facilitated diffusion and have very high apparent affinity constants for transport (>50 mM for lactose), which explains why low sugar concentrations do not protect against hyperosmotic conditions. We conclude that the more severe growth inhibition by salt stress than by equiosmolal concentrations of sugars reflects the inability of the cells to accumulate K+ (or Na+) to levels high enough to restore turgor as well as deleterious effects of the electrolytes intracellularly.     

The effects of anxiety and of situational stress on communicative efficiency.

Results of the study indicate that anxiety and stress interfere with a person's effectiveness of communication. 15 refs. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Cocaine-induced effects on isolation stress in neonatal rats.

The effects of cocaine administration on isolation-induced vocalizations and activity levels in 10-day-old rat pups were examined. Day 10 pups given cocaine (1.25, 2.5, 5, 10, and 20 mg/kg ip) vocalized significantly less than their caffeine- (10 mg/kg) and saline-administered siblings during a 5-min isolation period. Cocaine- and caffeine-administered pups also demonstrated a significant increase in overall activity compared with controls. In addition, ip administration of the dopamine antagonist haloperidol (0.5 and 1.0 mg/kg) before 1.25 and 2.5 mg/kg cocaine produced a significant elevation in vocalizations compared with saline pretreatment, which indicates a blocking of cocaine's effect on calling behavior. These results suggest that the endogenous dopamine system involved with reinforcement and reward may quell the stress of isolation in the infant rat. (PsycINFO Database Record (c) 2010 APA, all rights reserved)