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A method for the redox substoichiometric determination of thallium employing chlorate as an oxidant has been developed. The substoichiometric amount of Tl(III) formed was isolated by extraction with isoamyl acetate. 5-50 micrograms of thallium can be determined with an accuracy of +/- 1.3%.
The vitamin D binding protein (DBP) is a plasma protein that significantly enhances the chemotactic activity of C5a and C5a(desArg) (cochemotactic activity). The objective of this study was to investigate how DBP mediates this process using neutrophils and U937 cells transfected with the C5a receptor (U937-C5aR cells) and comparing chemotaxis to C-activated serum (DBP dependent) vs purified C5a (DBP independent). Binding to the cell surface is essential for this protein to function as a chemotactic cofactor, and DBP binds to a chondroitin sulfate proteoglycan (CSPG) on neutrophil plasma membrane preparations. To determine whether a CSPG also functions to mediate cochemotactic activity, U937-C5aR cells were grown in chlorate to inhibit CSPG sulfation or treated with chondroitinase AC. Either treatment significantly inhibited chemotaxis only to C-activated serum. CD44 is a major cell surface CSPG on leukocytes, and functions to facilitate chemotaxis. Treatment of cells with anti-CD44 blocks chemotaxis of neutrophils and U937-C5aR cells to C-activated serum but not purified C5a. DBP binds to CD44 on the cell surface as evidenced by coimmunoprecipitation, confocal microscopy, and cell binding studies. Annexin A2 associates with CD44 in lipid rafts; therefore, its potential role in mediating cochemotactic activity was investigated. Results demonstrate that anti-A2 inhibits neutrophil and U937-C5aR chemotaxis specifically to C-activated serum, blocks DBP binding to cells, and colocalizes with anti-DBP on the cell surface. These results provide clear evidence that CD44 and annexin A2 mediate the C5a chemotactic cofactor function of DBP.
The prevalence of pathogenic Yersinia enterocolitica in pig herds was monitored during six trials (at four different farrow-to-finisher farms). Samples were taken throughout the whole rearing period from birth of the piglets to the final fattening stage, and different samples were taken from these pigs during the slaughter process. Environmental samples also were evaluated to identify potential sources of on-farm infection. Y. enterocolitica was isolated using irgasan-ticarcillin-potassium chlorate broth enrichment and cefsulodin-irgasan-novobiocin agar culture. Colonies were identified using bio- and serotyping methods and by PCR assay. Pathogenic Y. enterocolitica were not isolated from fecal samples from piglets and weaners. The only fecal samples positive for Y. enterocolitica were obtained during the fattening stage. The prevalence of Y. enterocolitica in fattening pig herds ranged between 0 and 65.4%. Y. enterocolitica isolates were detected at the abattoir in 38.4% of the tonsils, in 3.8% of the ileocecal lymph nodes, on 0.3% of the carcass surfaces before chilling, and on 0% of the carcass surfaces after chilling. Almost all isolates belonged to bioserotype 4/O:3. Only one strain was identified as O:9. All isolates contained the ail gene. The yopT gene was found in 99.1% of the farm isolates but in only 76.6% of the isolates found at the abattoir from the corresponding carcasses. Although a direct link between porcine isolates and human infection has not been demonstrated, the similarity of the bioserotypes in infected pigs and humans and the presence of virulence factors in porcine isolates should encourage further studies to determine the risk of transmission of Y. enterocolitica to humans from pigs and pork products.
A national study was conducted for the isolation of pathogenic Yersinia enterocolitica in pig feces in the United States as part of the National Animal Health Monitoring System's Swine 2000 study. Fecal samples collected from swine operations from September 2000 to March 2001 from 77 production sites in 15 of the top 17 swine-producing states were tested for the presence of pathogenic Y. enterocolitica. After enrichment of swine fecal samples in irgasan-ticarcillin-potassium chlorate broth, the enriched cultures were plated on cefsulodin-irgasan-novobiocin agar for isolation of presumptive Y. enterocolitica. The isolates were confirmed as pathogenic Y. enterocolitica by the fluorogenic 5' nuclease PCR assay targeting the chromosomal attachment invasion ail gene. Of 2793 fecal samples tested, 106 (3.80%) ail-positive strains of Y. enterocolitica were isolated. These 106 ail-positive isolates originated from 7 of the 15 participating states. The predominant serotype O:3 (n = 79 of 106) was distributed in five states (n = 5 of 7). Serotype O:5 (n = 27 of 106) was also found in five states (n = 5 of 7). All isolates contained the virulence plasmid and expressed virulence-associated phenotypic characteristics. These results indicate that swine in the United Stares harbor Y. enterocolitica that can potentially cause human illness.
Between 1979 and 1983 less than 1% of admissions from acute poisoning in the UK were due to pesticides and fewer than 4% of admissions in those under 5 years were from this cause. Organochlorine, organophosphorus and carbamate insecticides account for only 10% of the total in both children and adults. Suspected pesticide poisoning was the cause of fewer than 0.3% of home accidents in those under 10 years of age and less than 4% of suspected poisonings documented by the Home Accident Surveillance System. Rodenticides were thought to be involved in 62% of these cases. Of children who presented to hospital 42% were admitted and 93% of these were discharged home within 2 days. In the UK, the morbidity from acute pesticide poisoning in children is low and the mortality is nil and there is therefore no evidence to support the view that paediatric pesticide intoxication is a significant clinical problem. Though no fatalities were recorded in children, pesticides were responsible for 1.3% of all deaths due to poisoning in the UK between 1979 and 1983. In adults admitted to hospital, the mortality from pesticide poisoning is approximately 12% and three quarters of these deaths are due to the deliberate ingestion of paraquat. The general term pesticide refers to a group of products that are used as insecticides, acaricides, fungicides, herbicides, rodenticides, and plant growth agents. Chemically, the group includes bipyridilium compounds, carbamates, chloralose, chlorates, coumarins, dinitro compounds, dithiocarbamates, fluoroacetates, organochlorine organophosphorus and organotin compounds, pentachlorophenol, phenoxyacetates, phosphine (as magnesium and aluminium phosphides), pyrethrins, pyrethroids and triazines.(ABSTRACT TRUNCATED AT 250 WORDS)
A dependence on proteoglycans for cationic lipid-mediated gene transfer has been suggested in previous studies [Mislick and Baldeschwieler (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 12349-12354; Mounkes, Zhong, Cipres-Palacin, Heath and Debs (1998) J. Biol. Chem. 273, 26164-26170]. We have evaluated the mechanism of proteoglycan involvement in cationic lipid-mediated gene transfer. DNA plasmid uptake and gene expression were studied in wild-type Chinese hamster ovary (CHO) cells (CHO-K1), heparan sulphate-deficient CHO cells (pgsD-677) and proteoglycan-deficient CHO cells (pgsB-618). At an optimal ratio of cationic lipid to DNA, a substantial decrease in reporter gene expression was observed in proteoglycan-deficient cells compared with that in heparan sulphate-deficient and wild-type cells. However, there were no differences in reporter gene expression between the cell lines when transfected by electroporation. Moreover, all cell lines exhibited equal cationic-lipid-DNA complex uptake activities, as assessed by the measurement of intracellular (32)P-labelled and rhodamine-labelled DNA plasmid. An analysis of reflected-light images of wild-type and proteoglycan-deficient cells suggested that cationic lipids were preferentially toxic to proteoglycan-deficient cells. Cell-growth assays confirmed this, showing that cationic lipids exhibited a greater anti-proliferative activity in proteoglycan-deficient cells and in chlorate-treated wild-type cells than in the other cell lines. The growth-inhibitory effect of cationic lipids was abrogated by the addition of exogenous sulphated glycosaminoglycans. We conclude that the glycosaminoglycan part of proteoglycans serves a protective role against cationic lipid cytotoxicity, allowing optimal transfection efficiency in vitro.
Pesticide intoxications are mainly accidental with a benign course, such as ingestions of diluted fertilizers or low concentration antivitamin K rodenticides, ant-killing products or granules of molluscicides containing 5% metaldehyde. Voluntary intoxications by chloralose, strychnine, organophosphorus or organochlorine insecticides, concentrated anti-vitamin K products, herbicides such as paraquat, chlorophenoxy compounds, glyphosate or chlorates may be severe. Toxicity is due to active substances but also to solvents or surfactants included in the composition. Analysis done in a toxicology laboratory help in establishing diagnosis, prognosis and treatment. Poison centres may be called constantly to help for the evaluation of these intoxications.
The detection and identification of traces of inorganic ions and sugars can play a major role in the forensic investigation of an explosives related incident. This survey investigated the background levels of these substances in the general environment. Six sampling locations were selected from around the mainland of the United Kingdom, representing urban and rural sites. Swab and vacuum samples were collected from different locations within each site including motor vehicles, private houses, hotels, the exterior of buildings, road surfaces, and street signs. Sampling was carried out in summer and winter to investigate changes in the levels of the target species due to seasonal factors such as road treatments or weather. The samples were extracted with water and analyzed for a range of inorganic anions, cations, and sugars using ion chromatography. Most of the target anions were found to be common to all locations. Chloride, sulphate, nitrate, and phosphate were found to be the most common and the most abundant. Chlorate was found at a low level in some external samples. Perchlorate and thiocyanate were not detected in any samples. There was a marked increase in the quantity of sodium and chloride detected in samples collected during the winter. Sodium and calcium were detected in most samples. Potassium and magnesium were detected in approximately half of the samples. Ammonium was less common but detected at significant levels in wall samples. Glucose, fructose, and sucrose were detected in the vacuum samples from the interior surfaces of houses, hotels, and cars.
The soluble fraction of the rat and rabbit renal homogenate (normal, after acute blood loss or cobalt chlorate administration) contains an erythropoietin inhibitor and interferes with transformation of the renal erythropoiesis--stimulating factor into erythropoietin. After subcutaneous administration of this fraction in intact rats, an erythropoiesis inhibitor was revealed in their plasma.
An heterologous transformation system for entomopathogenic fungi B. bassiana and M. anisopliae was developed based on the use of A. nidulans nitrate reductase gene (niaD). B. bassiana and M. anisopliae niaD stable mutants were selected by treatment of protoplast with ethane methane sulphonate (EMS) and regenerated on chlorate medium. The cloned gene was capable of transforming B. bassiana and M. anisopliae at a frequency of 5.8 to 20 transformants per microg of DNA. Most of them were mitotically stable.
A novel, simple, rapid, sensitive and highly selective flow injection procedure for the spectrophotometric determination of chlorine dioxide in the presence of other chlorine species, viz, free chlorine, chlorite, chlorate and hypochlorite, is developed. The method is based on the discoloration reaction between chlorine dioxide and chlorophenol red and can overcome the shortcomings existed in direct spectrophotometric determination for chlorine dioxide owing to the serious interference of free and combined chlorine. The procedure gave a linear calibration graph over the range 0-0.71 mg/L of chlorine dioxide. With a detection limit of 0.024 mg/L and a sample throughput of 60 samples/h.
A simple sample preconcentration technique for the determination of trace level of bromate and perchlorate in drinking water with ion chromatography is presented. With the hydrophobic anion-exchange column and a sodium hydroxide eluent in linear gradient, bromate and perchlorate can be determined in a single injection within 35 min. Prior to ion chromatographic analysis, the drinking water sample was treated with the OnGuard Ag cartridge to remove the superfluous chloride and concentrated in 20-fold with poly(tetrafluoroethylene)(PTFE) beaker in a domestic microwave oven in 15 min. The recoveries of the analytes are better than 90%. The detection limits for bromate, perchlorate, iodate and chlorate were 0.10 microgram/L, 0.20 microgram/L, 0.11 microgram/L and 0.21 microgram/L, respectively. The developed method is applicable for the quantitation of bromate and perchlorate in real drinking water samples.
The amyloid protein precursor (APP) of Alzheimer's disease was found to bind saturably (Kd = 60 nM) to embryonic chick brain extracellular matrix (ECM). The binding of APP to ECM was not inhibited by 10 micrograms/ml heparin or heparan sulfate. However, pretreatment of cells with 1 mM 4-methylumbelliferyl-beta-D-xyloside, an inhibitor of proteoglycan biosynthesis, reduced the number of APP binding sites on the ECM by 80%. The binding of APP to ECM was also inhibited by pretreatment with chlorate, an inhibitor of glycan sulfation, and heparitinase, which digests the carbohydrate component of heparan sulfate proteoglycans. These results suggest that APP binds with high affinity to one or more heparan sulfate proteoglycans. Acidic and basic fibroblasts growth factor (FGF) also bound to chick ECM. When ECM was incubated with a protease associated with the enzyme AChE (AChE-AP), APP and acidic FGF were released intact from the matrix. The AChE-AP was at least 100-fold more potent in releasing APP from ECM than other trypsin-like proteases (trypsin, plasmin, thrombin). The action of the AChE-AP was inhibited by glia-derived nexin (protease nexin I) and by human brain APP at low nanomolar concentrations. These results suggest that in vivo an AChE-AP may cleave ECM proteins to regulate the availability of soluble APP or other factors bound to the ECM.
OBJECTIVE: To characterize the associations and functions of apoE with the HepG2 cell surface through the study of the relationship between proteoglycans and apoE on HepG2 cell surface. METHODS: The specific binding of 7C9, a monoclonal antibody against the N-terminal domain of apoE was employed to demonstrate the effects of glycosaminoglycans, heparinase, chondroitinase, xyloside, and chlorate on the apoE of HepG2 cell surface. RESULTS: Growth of cells in beta-D-xyloside decreased cell surface apoE by 45% with a concomitant increase in apoE secretion (4.3-fold), underlining the importance of glycosaminoglycan association of apo E. Incubations with heparinase (3 U/ml) and heparin (1 mg/ml) decreased apoE by 25.0% and 30.5% respectively indicating association through cell surface haparin sulfate proteoglycans. Incubations with chondroitinase ABC (1.5 U/ml) reduced cell surface apoE by 40.0%. Chondroitin sulfate A and chondroitin sulfate B reduced cell surface apoE by 23.6% and 15.3% respectively while incubations with chondrointin sulfate C not effective. Decreasing the levels of cell surface apoE with haparin or chondrointin sulfates A and B increased the subsequent binding of LDL to the HepG2 cell surface. CONCLUSIONS: ApoE associates with the cell surface mainly through chondrointin sulfate proteoglycans and to a lesser extent through heparan sulfate proteoglycans, decreased levels of cell surface apoE increase the binding of LDL to the cell surface.
A sensitive selective method of unsaturated chlorated hydrocarbons detection by means of gas chromatography in skin was worked out. The limits are: for 3,4-dichlorobutene (DCB)-1-0.0024 mu g, for 1,3 DCB-2-0.0027 mu g, for 1,4-DCB-2 (cys)-0.0024 mu g, for 1,4-DCB-2 (trans)-0.0032 mu g, for 2,3,4 trichlorobutene (TCB)-1-0.0093 mu g in analysed volume. The method can be used for the sanitary control of the skin contamination in subjects exposed to DCB and TCB.
The plasmid RP4::Mu cts62 in stably inherited by Erwinia carotovora 268 strain. Under the conditions of thermoinduction bacteriophage Mu is segregated and completely eliminated more intensively than in Escherichia coli cells. At thermoinduction the transposition of bacteriophage Mu cts62 into different chromosomal sites takes place, causing the induction of chlorate resistant and auxotrophic mutants with the frequency of 10(-4). Two clones deficient in production of 2 of the 4 resident prophages of Erwinia carotovora 268 strain were found among Mu-induced mutants. The deleted prophages are E105 and 59. DNA-DNA hybridization has revealed the complete and partial deletions of bacteriophage E105 with the level of L-asparaginase production in the cells remaining intact. The damage of the prophage 59 is probably caused by point mutations or short deletions.
Basic fibroblast growth factor (bFGF) was internalized at a rapid rate by Chinese hamster ovary (CHO) cells that do not express significant numbers of high affinity receptors for bFGF as well as CHO cells that have been transfected with cDNA encoding FGF receptor-1 or FGF receptor-2. Internalization of bFGF was completely blocked by the addition of 10 micrograms/ml heparin in the parental CHO cells but only partially inhibited in cells expressing transfected FGF receptors. Bovine aortic endothelial cells also exhibit heparin-sensitive and heparin-resistant internalization of bFGF. The internalization of bFGF through the heparin-resistant pathway in CHO cells was efficiently competed by addition of unlabeled bFGF, was proportional to the number of receptors expressed, and approached saturation, suggesting that the heparin-resistant internalization was due to high affinity receptors. Internalization of bFGF through the heparin-sensitive pathway was not efficiently competed by unlabeled bFGF and did not approach saturation at concentrations of bFGF up to 50 ng/ml, properties similar to the interaction of bFGF with low affinity heparan sulfate binding sites on the cell surface. Internalization of bFGF in CHO cells not expressing FGF receptors was inhibited by heparin, heparan sulfate, and dermatan sulfate, the same glycosaminoglycans that block binding to cell-surface heparin sulfates. Internalization of bFGF in the parental CHO cells was inhibited at the same concentrations of heparin that block binding to cell-surface heparan sulfates. Finally, inhibition of the sulfation of CHO cell heparan sulfates by the addition of chlorate or digestion of CHO cell heparan sulfates with heparinase inhibited bFGF internalization in the parental CHO cells. These results demonstrate that bFGF can be internalized through a direct interaction with cell-surface heparan sulfates. Thus, there are two pathways for internalization of bFGF: high affinity receptor-mediated and heparan sulfate-mediated.
BACKGROUND: Survival time within a disabled submarine (SUBSUNK) is dependant on atmospheric composition and proper design and use of emergency atmospheric control systems. The objective of this study was to investigate atmospheric changes and physiological responses during a SUBSUNK trial. METHODS: There were 18 volunteers who were restrained within a 250 m3 front compartment of an Ula-class submarine submerged in 8 degrees C seawater for 6 d, 18 h. Atmospheric control was maintained according to emergency procedures using non-electrically powered chemical CO2 absorption, and O2 was replenished using chlorate candles. Atmospheric parameters, skin and body temperatures, weight, urine, and drinking volume were measured. Subjective responses to cold were measured on a visual analog scale (VAS), and symptoms were logged on the environmental symptoms questionnaire (ESQ). RESULTS: Atmospheric temperature gradually decreased to a minimum of 14.1 degrees C. Toe, heel, and finger temperatures decreased significantly. Subjects reported inferior subjective thermal comfort on the VAS and increased cold stress on the ESQ. Except for CO2, no inorganic or volatile organic compounds exceeded occupational exposure limits. The PO2 and PCO2 ranged from 17.4-20.3 and 1.9-2.8 kPa, respectively, during the first 5 d. During the last 2 d, PO2 and PCO2 were deliberately maintained at about 15.8 and 3.1 kPa, respectively. Mean oxygen consumption and CO2 production were 23.8 and 19.8 L standard temperature and pressure (STP) x man(-1) x h(-1), respectively. Soda lime and lithium hydroxide CO2 absorption capacities were 126 and 405 L STP x kg(-1) respectively. CONCLUSIONS: Atmospheric conditions can be controlled acceptably for 6 d, 18 h within the front compartment of an Ula-class submarine operating according to emergency SUBSUNK procedures.