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PubMed2002

Primary structure and catalytic properties of a cold-active esterase from a psychrotroph, Acinetobacter sp. strain No. 6. isolated from Siberian soil.

We cloned a gene coding for a cold-active esterase from a genomic library of Acinetobacter sp. strain No. 6, a psychrotroph isolated from Siberian soil. The gene, aest, encoded a protein of 301 amino acid residues, the deduced sequence of which had less than 17% identity to sequences of known esterases and lipases. However, the esterase seemed to belong to the alpha/beta hydrolase superfamily, because it contained a sequence, Gly-Xaa-Ser-Xaa-Gly (with Xaa an arbitrary amino acid residue), found in most serine hydrolases of this superfamily. Sequence comparison earlier suggested a weak phylogenetic relationship of gene product AEST to the EST group of the esterase-lipase family, which has been found only in eukaryotes. The aest gene was expressed in Escherichia coli BL21(DE3) cells under the control of the T7 promoter, and the expression product was purified to homogeneity and characterized. It catalyzed the hydrolysis of esters with short-chain acyl groups and had lower activation energy and lower thermostability than do mesophilic enzymes, as expected from the cold-adapted nature of this enzyme.

Acinetobacter↗
PubMed1976

[Short term effects of diets with high levels of dithiocarbamates on carbohydrate and lipid metabolism of rat liver].

The purpose of the work was to establish the eventual "metabolic toxicity" of pesticide-contaminated diets in the Rat. The liver metabolic response to various stimuli was compared in dithiocarbamate-fed animals and in non-contaminated ones. 112 weanling male Wistar CF rats were fed, during 15 days, with a demi-synthetic control diet. They were then divided into 4 lots:--the control group C, which went on to receive the same diet,--the nabame group N, the diet of which was supplemented with 275 ppm of the dithiocarbamate;--the thirame groupe R, receiving the control diet + 600 ppm thirame;--the zineb groupe Z, given the control diet + 3 600 ppm Zineb. The animals were fed with these diets during 14 days, their dithiocarbamate intake thus averaging 1/20 th of the per os LD 50/rat/day. At the end of this 2-week period, each of the 4 groups was divided into 4 sub-groups, all the animals were fasted overnight, then sacrified:--after no other treatment (sub-groups T);--30 minutes after an i.p. injection of 2.6 g/kg glucose (G);--after having been forced to walk in a restraint wheel for 50 minutes/hr during the 18 hrs of the night fast (sub-groups W);--after a 90 minutes exposure in a cold room (F). The weights of the animals, of their liver, heart, kidneys, adrenals and epididymal pads were recorded. In their liver, the following compounds were determined: water, proteins, total lipids, triglycerides, long-chain acyl-CoA, non-esterified fatty acids, total cholesterol, glycogen, glucose, alpha-glycerophosphate. The thirame rats had a lower food intake than the others and the smallest body weight, but their relative liver and kidneys weights were the highest. The nabame animals did not differ from the control ones but the zineb rats had the lightest epididymal fat pads. The primary effects of the dithiocarbamate diets on liver metabolism were apparently not the same in the 3 groups compared to the control ones: nabame and thirame increased glycogen, thirame increased the lipid compounds: long-chain actyl-CoA and triglycerides, where as zineb feeding resulted in an increase of glucose concentration and in a decrease of triglycerides and total lipids. Muscular exercise, or cold exposure, had the following effects compared to those they had in the control group: a greater glucose utilization in the nabame and thirame rats, a smaller glycogen and glucose utilization, associated with an increase of alpha-glycerophosphate, in the zineb animals. These results were considered altogether with those obtained in a previous paper by the same authors, which concerned liver ketone bodies and adenine nucleotides changes after the same experimental conditions, and it was concluded that the 3 dithiocarbamates actually had a common effect on rat metabolism: they all impaired glucose utilization by the liver. Also, fat mobilization from peripheral depots was shown to occur in the 3 experimental groups, resulting in liver fatty acid oxidation in the nabame and zineb rats, and in liver steatosis for the thirame ones...

Adipose Tissue↗
PubMed2000

The S. pombe rlc1 gene encodes a putative myosin regulatory light chain that binds the type II myosins myo3p and myo2p.

In order to identify additional components important for cell division in the fission yeast Schizosaccharomyces pombe we have screened a bank of conditional cold-sensitive mutants for cytokinesis defects. One of these mutants showed a delay in cell cleavage, and strong genetic interactions with other genes implicated in medial ring formation. Cloning of the corresponding gene indicates that it encodes a protein with significant homology to the regulatory light chain of non-muscle myosins. We have named the gene rlc1 (regulatory light chain 1). The gene is not essential for division, but null mutants display a cell cleavage defect and form an aberrant F-actin ring. Two myosin-II heavy chains have been identified in fission yeast: Co-immunoprecipitation experiments indicate that rlc1p associates more strongly with myo3p than myo2p.

Amino Acid Sequence↗
PubMed1986

Finger peripheral resistance during local cold provocation in vasospastic disease.

Finger systolic blood pressure (FSBP), finger arterial inflow (FAI), finger venous opening pressure (FVOP), and finger peripheral resistance (FPR) were evaluated in 56 workers exposed to chain-saw vibration. Twenty-one of the workers were free from vibration-induced white finger (VWF). In 17 subjects VWF had ceased; 12 of the subjects had active VWF; 6 subjects had Raynaud's disease. The subjects were examined in a supine position under thermoneutral conditions with strain-gauge plethysmography. Cold provocation of the finger caused a significant reduction in FSBP in the groups with Raynaud's disease and active VWF. The finger circulation of the subjects with active VWF and that of those with Raynaud's disease showed a significantly reduced FAI when compared with that of the symptom-free referents. Finger cooling produced a gradual reduction in the FVOP and a significantly increased FPR of the subjects with active VWF and in those with Raynaud's disease. FPR was already increased in the Raynaud's disease group at the beginning of the test, whereas it increased in the VWF group as the temperature fell. The results suggest that in Raynaud's disease the fault is in the level of activity of the sympathetic outflow and in VWF it is peripheral mechanisms controlling the vessel tone.

Cold Temperature↗
PubMed2001

Role of glutathione in adaptation and signalling during chilling and cold acclimation in plants.

Glutathione is an important component of the ascorbate-glutathione cycle, which is involved in the regulation of hydrogen peroxide (H2O2) concentrations in plants. During chilling and cold acclimation, i.e. exposure to temperatures between 0 and 15 degrees C, H2O2 may accumulate. Excess electrons from the photosynthetic and respiratory electron transport chains can be used for the reduction of oxygen, thus producing superoxide radicals (O2.-); these are subsequently transformed to H2O2 via superoxide dismutase (SOD; EC 1.15.1.1). During the removal of excess H2O2, reduced glutathione (GSH) is converted to its oxidised form (GSSG), and GSH is regenerated by the activity of NADPH-dependent glutathione reductase (GR; EC 1.6.4.2). At low non-freezing temperatures, high GSH content and GR activity were detected in several plant species, indicating a possible contribution to chilling tolerance and cold acclimation. Changes in H2O2 concentration and GSH/GSSG ratio alter the redox state of the cells and may activate special defence mechanisms through a redox signalling chain. The finding that several defence genes have antioxidant responsive elements or GSSG binding sites in their regulatory regions supports the idea that redox signalling is involved in regulating gene expression in response to low temperature.

Journal Article↗
PubMed1990

The effects of very low fat diets enriched with fish or kangaroo meat on cold-induced vasoconstriction and platelet function.

Eighteen healthy volunteers consumed very low fat diets (less than 7% of daily energy) enriched with different sources of long chain (C20 and C22) polyunsaturated fatty acids (PUFA). Three diets provided 500 g/day of fish caught in the tropical waters of Australia (rich in arachidonic acid and docosahexaenoic acid), fish caught in the southern waters of Australia (rich in docosahexaenoic acid), or kangaroo meat (rich in linoleic and arachidonic acids). The fourth diet was vegetarian, similarly low in fat but containing no 20- and 22-carbon PUFA. An increase in the percentage of a particular C20 or C22 PUFA in the plasma phospholipid fraction in subjects consuming these low fat diets corresponded to the dietary PUFA composition. This study examined the effect of dietary modification of the level of arachidonic acid in plasma phospholipids on both traditional measures of platelet function and on cold-induced vasoconstriction. The cold pressor response, measured by venous occlusion plethysmography, was depressed in diets which elevated the levels of arachidonic acid in plasma lipids (kangaroo and tropical fish), enhanced after subjects consumed a diet which increased the levels of docosahexaenoic acid and eicosapentaenoic acid (southern fish diet), and was unchanged by the low fat vegetarian diet. There was no effect on bleeding time or platelet responsiveness.

Adult↗
PubMed1992

Role of sympathetic innervation in brown adipocyte proliferation.

The role of sympathetic nerves in the control of brown adipose tissue (BAT) growth was studied by quantitative photonic autoradiography using [3H]-thymidine to follow mitotic activity. The effects of cold exposure (4 days at 5 degrees C) and chronic norepinephrine treatment (0.375 mumol/h for 4 days at 25 degrees C) were compared in intact and denervated interscapular BAT pads of adult rats. The denervated pads were paler than the contralateral innervated sides and contained numerous adipocytes filled with large lipid droplets. In the innervated BAT pad, cold exposure markedly enhanced the labeling frequency in brown adipocyte precursor cells (interstitial cells and preadipocytes) and in endothelial cells forming the numerous blood capillaries (P < 0.01). However, in the denervated BAT pads, the effects of cold exposure on cell proliferation were markedly inhibited. Remarkably, norepinephrine infusion at 25 degrees C mimicked the effects of cold exposure on the coordinated proliferation of the different cellular types, both in the innervated and denervated pads. These results demonstrate that intact sympathetic nerves are required for the stimulation of mitotic activity in BAT by cold exposure but not by norepinephrine infusion. It appears therefore that norepinephrine release from sympathetic nerves represents the initial stimulus in a chain of metabolic events leading to BAT hyperplasia. However, sympathetic or sensory nerves may still contain factors modulating cell proliferation and differentiation in BAT.

Adipose Tissue, Brown↗
PubMed2000

An enzyme-linked lectin assay for alpha1,3-galactosyltransferase.

UDP-Gal:Galbeta1-4GlcNAc alpha1,3-galactosyltransferase (alpha3GalT) is responsible for the synthesis of carbohydrate xenoantigen Galalpha1-3Galbeta1-4GlcNAc. In this work a convenient and sensitive assay system for quantification of alpha3GalT activity by enzyme-linked lectin assay (ELLA) with colorimetric detection is described. Microtiter plate wells whose surface had been coated with the polyacrylamide conjugate of the disaccharide Galbeta1-4GlcNAc (acceptor) are incubated with alpha3GalT in the presence of "cold" UDP-Gal as glycosyl donor. Formation of product by enzymatic extension of the glycan chain is detected by the biotinylated plant lectin Viscum album agglutinin. The standard curve for correct quantification of alpha3GalT activity is completed after running standard assays with no (background) or known quantities of enzyme activity. Product formation detected in this manner is proportional to enzyme activity and the concentrations of the acceptor and the glycosyl-donor UDP-Gal. In accordance with the known specificity of alpha3GalT, no enzymatic conversion of Le(x) into GalalphaLe(x) was observed using this assay. Human alphaGal antibodies were isolated using a disaccharide-exposing affinity adsorbent and their specificity was studied. Relative to the application of these natural immunoglobulins as product-detecting tool, the ELLA proved to be more sensitive.

Animals↗
PubMed1994

The role of the thymus in myasthenia gravis.

The experimental work discussed here supports the hypothesis that in the pathogenesis of MG the initial and essential steps take place within the thymus. Most if not all thymuses of MG patients contain B cells capable of producing AChR specific autoantibody along with appropriate stroma elements. Hyperplastic thymuses characteristically contain germinal centers with cellular complexes of AChR-producing MC and surrounding interdigitating dendritic cells. In thymomas, the source of the myasthenogenic autoantigen is less obvious. There are data suggesting that thymoma epithelium expresses a protein sharing certain peptide epitopes with the AChR alpha chain, although there is no further molecular similarity. A unique type of 'molecular self-mimicry' cold be involved in the initiation of thymoma-associated MG.

Animals↗
PubMed2006

Prevalence of viral respiratory tract infections in children with asthma.

BACKGROUND: Previous studies support a strong association between viral respiratory tract infections and asthma exacerbations. The effect of newly discovered viruses on asthma control is less well defined. OBJECTIVE: We sought to determine the contribution of respiratory viruses to asthma exacerbations in children with a panel of PCR assays for common and newly discovered respiratory viruses. METHODS: Respiratory specimens from children aged 2 to 17 years with asthma exacerbations (case patients, n = 65) and with well-controlled asthma (control subjects, n = 77), frequency matched by age and season of enrollment, were tested for rhinoviruses, enteroviruses, respiratory syncytial virus, human metapneumovirus, coronaviruses 229E and OC43, parainfluenza viruses 1 to 3, influenza viruses, adenoviruses, and human bocavirus. RESULTS: Infection with respiratory viruses was associated with asthma exacerbations (63.1% in case patients vs 23.4% in control subjects; odds ratio, 5.6; 95% CI, 2.7- 11.6). Rhinovirus was by far the most prevalent virus (60% among case patients vs 18.2% among control subjects) and the only virus significantly associated with exacerbations (odds ratio, 6.8; 95% CI, 3.2-14.5). However, in children without clinically manifested viral respiratory tract illness, the prevalence of rhinovirus infection was similar in case patients (29.2%) versus control subjects (23.4%, P > .05). Other viruses detected included human metapneumovirus (4.6% in patients with acute asthma vs 2.6% in control subjects), enteroviruses (4.6% vs 0%), coronavirus 229E (0% vs 1.3%), and respiratory syncytial virus (1.5% vs 0%). CONCLUSION: Symptomatic rhinovirus infections are an important contributor to asthma exacerbations in children. CLINICAL IMPLICATIONS: These results support the need for therapies effective against rhinovirus as a means to decrease asthma exacerbations.

Adolescent↗
PubMed2003

Ileal cytokine dysregulation in experimental necrotizing enterocolitis is reduced by epidermal growth factor.

BACKGROUND: Necrotizing enterocolitis (NEC) is the most common gastrointestinal disease of premature infants. We have shown in previous studies that proinflammatory interleukin-18 and interleukin-12 are up-regulated in the ileum of rats with experimental NEC and that epidermal growth factor (EGF) reduces the development of disease. Here we investigated whether the protective effects of EGF are a result of changes in ileal interleukin-18, interleukin-12 and/or antiinflammatory interleukin-10. METHODS: Newborn rats were artificially fed with either growth-factor-free rat milk substitute (RMS) or RMS supplemented with 500 ng/mL EGF (RMS + EGF) and NEC was induced via exposure to asphyxia and cold stress. Cytokine expression and localization were assessed using reverse-transcription real-time polymerase chain reaction and immunohistology/confocal microscopy. RESULTS: Enteral administration of EGF (RMS + EGF) decreased overproduction of interleukin-18 and increased interleukin-10 production in the ileum. Furthermore, increased interleukin-10 production was associated with up-regulation of the transcription factor Sp1 in RMS + EGF rats. CONCLUSIONS: These data suggest that EGF may reduce NEC via increased interleukin-10 and decreased interleukin-18 and that EGF-mediated up-regulation of Sp1 may account for the increased interleukin-10.

Animals↗
PubMed1988

High-titer, high-thermal-amplitude cold autoagglutinin not associated with hemolytic anemia.

An example of an unusual cold autoagglutinin is reported. The antibody was monoclonal IgMKappa able to fix complement, and, in the presence of albumin, had both a high titer (greater than 4,096 at 4 degrees C) and a wide thermal range (4-37 degrees C). The patient was closely followed over a 3-year period with no evidence of hemolysis ever documented, despite a persistently positive direct antiglobulin test and the presence of the cold autoagglutinin. In contrast to previous reports regarding cold-agglutinin disease, this case demonstrates that in vivo hemolysis is not always associated with cold autoagglutinins that in vitro show a high thermal range in the presence of albumin.

Agglutinins↗
PubMed1998

Influence of cold stress on the preliminary enrichment time needed for detection of enterohemorrhagic Escherichia coli in ground beef by PCR.

The influence of cold stress at 4 and 0 degree C on the detection time as assessed by impedance technology (Bactometer; Biomérieux, Marcy l'Etoile, France) of different enterohemorrhagic Escherichia coli (EHEC) strains was determined. Although there is some variation in susceptibility among EHEC strains, prolonged exposure of EHEC to cold stress, i.e., 4 and 5 days at 4 and 0 degree C, respectively, in general significantly increased their detection time. This reflects an increase of the lag-phase time caused by cold stress. Two EHEC strains were selected to determine the minimum preliminary enrichment time that would ensure a positive PCR detection of low numbers of verotoxin-producing E. coli (VTEC; 2 to 2 x 10(5) CFU/25 g) inoculated into ground beef (25 g) and stored at 4 or -20 degrees C for 8 and 14 days, respectively. Incubation times of 6 and 9 h of 1 to 10 CFU/g and 1 to 10 CFU/25 g, respectively, were sufficient for PCR detection of VTEC in ground beef when analysis was performed immediately after inoculation (no cold stress). When cells are exposed to cold stress (4 or -20 degrees C) a 24-h enrichment period is recommended. Restriction of enrichment time to 9 h under these circumstances decreases the sensitivity of PCR detection to 80 CFU/g. Hence, to obtain maximum sensitivity, PCR detection of VTEC in naturally contaminated ground beef should be performed after 24 h of enrichment.

Animals↗
PubMed1995

Natural and experimental rhinovirus infections of the lower respiratory tract.

Rhinoviruses are the cause of the majority of common colds, but their role in lower respiratory disorders is less clear. Recent studies using the polymerase chain reaction to detect rhinoviruses have established respiratory viral infections as major factors in the induction of acute exacerbations of asthma in both adults and children, both in mild exacerbations and in more severe exacerbations leading to hospital admission. Rhinoviruses were the major virus type detected in these studies, accounting for two-thirds of viruses detected. It is not known whether rhinoviruses produce their effects by directly invading the lower airway or by indirect means. Previous clinical studies provide some evidence that rhinoviruses are capable of infecting the lower airway. However, the immunologic response, both in the upper and lower airways, remains poorly defined. Recent studies have provided evidence of increased cellular activation in peripheral blood and in bronchial biopsies in atopic subjects compared with normal subjects during experimental rhinovirus infections. The reasons for these different cellular responses are unclear. Rhinoviruses as well as other respiratory viruses have been shown to increase levels of a variety of cytokines from respiratory epithelium, monocytes, or macrophages. Prominent among these cytokines is interleukin (IL)-8. We have detected increased levels of IL-8 in nasal secretions from subjects with wild-type rhinovirus infections. We studied the mechanisms of rhinovirus-induced IL-8 production and and found protein release from both pulmonary epithelial and peripheral blood mononuclear cells. This protein production was accompanied by increased mRNA expression and evidence of infection of both pulmonary epithelial and monocyte cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1990

Suspected vibration magnitude on the body of chain saw users among National Forest workers in Kyusyu, from 1956 to 1984.

Chain saws were first used in Kyusyu in 1956 for cutting down trees and sawing timber in the national forests. In 1961, some workers complained of symptoms due to the vibrations, but countermeasures were not taken until later. The working system and payment system in those times were different from the systems, today. The magnitudes of the chain saw vibrations were very large and the chain saws were operated for many hours, each day. Workers with VWF (vibration induced white finger) had to continue to operate the chain saws without restriction. Vibration disease is related primarily to the vibration of the tools, but also to the transmission of the vibration to the workers. From the official data on chain saws used in national forests, vibration transmission to the body was determined for workers from 1956 to 1984 by comparing the vibration magnitude on the body during work with modern chain saws and during vibration loading tests with an electrodynamic shaker in the laboratory. The vibrations transmitted to the body in some rule in the observation. If the working posture of chain saw operator was the same as today, the vibration transmission was determined from the tool vibration, tool weight, joint fixation and orientation of the hands and the hands and the arms to the handle (joint elasticity). Older chain saws had 20 to 30 times larger vibration magnitudes than modern chain saws, and were also 2 to 3 times heavier. The vibrations transmitted to the foreheads of earlier workers were approximately the same magnitude as the vibrations at the back of the hands of present workers. Also the daily and annual hours of chain saw use were very long, and there were not sufficient measures to protect from the cold. The vibration in the former days led to more symptoms and lesions than in present times.

Elasticity↗