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PubMed1978

Resistance relationships in Mycobacterium smegmatis ATCC 607 to phages sensitive or resistant to both chloroform and streptomycin sulphate.

Four of eight mycobacteriophages did not form plaques after they were exposed to chloroform. Phages sensitive to chloroform did not produce plaques when plated on media containing 1000 microgram/ml of streptomycin sulphate. The same concentration of dihydrostreptomycin sulphate did not interfere with plaque formation. Mutants of Mycobacterium smegmatis resistant to each of the eight phages were isolated. Sensitivity or resistance to chloroform and streptomycin sulphate and phage resistant bacterial mutants may provide a basis for classifying the mycobacteriophages.

Adsorption↗
PubMed2006

Effects of aqueous, methanolic and chloroform extracts of rhizome and aerial parts of Valeriana officinalis L. on naloxone-induced jumping in morphine-dependent mice.

In the present study, the effects of rhizomes and aerial parts extracts of Valeriana officinalis L. on morphine dependence in mice have been investigated. Animals were treated subcutaneously with morphine (50, 50 and 75 mg/kg) three times daily (10 am, 1 pm and 4 pm) for 3 days, and a last dose of morphine (50 mg/kg) was administered on the fourth day. Withdrawal syndrome (jumping) was precipitated by naloxone (5 mg/kg) which was administered intraperitoneally 2 hours after the last dose of morphine. To study the effects of the aqueous, methanolic and chloroform extracts of both aerial parts and rhizome of the V. officinalis L. on naloxone-induced jumping in morphine-dependent animals, 10 injections of morphine (three administrations each day) for dependence and a dose of 5 mg/kg of naloxone for withdrawal induction were employed. Intraperitoneal injection of different doses (1, 5, 25 and 50 mg/kg) of aqueous, methanolic and chloroform extracts of the rhizome of V. officinalis L. 60 minutes before naloxone injection decreased the jumping response dose-dependently. Pre-treatment of animals with different doses (1, 5, 25, 50 and 100 mg/kg) of aqueous and methanolic extracts of aerial parts of V. officinalis L. 60 minutes before naloxone injection caused a significant decrease on naloxone-induced jumping. The chloroform extract of the aerial parts of V. officinalis L. did not show any significant changes on jumping response in morphine-dependent animals. It is concluded that the extracts of V. officinalis L. could affect morphine withdrawal syndrome via possible interactions with inhibitory neurotransmitters in nervous system.

Animals↗
PubMed1990

Routes of chloroform exposure and body burden from showering with chlorinated tap water.

While there is an awareness of the need to quantify inhalation exposure from showers, the potential for dermal exposure to organic contaminants in showers has not been appreciated or explored. To establish routes of environmental exposure from showers, comparisons of the concentration of chloroform in exhaled breath after a normal shower with municipal tap water were made with those after an inhalation-only exposure. The postexposure chloroform breath concentrations ranged from 6.0-21 micrograms/m3 for normal showers and 2.4 to 10 micrograms/m3 for inhalation-only exposure, while the pre-exposure concentrations were all less than the minimum detection limit of 0.86 micrograms/m3. According to an F-test, the difference between the normal shower and the inhalation-only exposures was considered significant at a probability of p = 0.0001. Based on the difference, the mean internal dose due to dermal exposure was found to be approximately equal to that due to the inhalation exposure. The effect of the showering activities on the concentration of chloroform shower air was examined by comparing air concentrations during a normal shower with the air concentrations obtained when the shower was unoccupied. The F-test showed that there is no significant difference between the two sets of data.

Air Pollutants↗
PubMed1990

Chloroform exposure and the health risk associated with multiple uses of chlorinated tap water.

Recently, showers have been suspected to be an important source of indoor exposure to volatile organic compounds (VOC). The chloroform dose to an individual from showering was determined based on exhaled breath analysis. The postexposure chloroform breath concentration ranged from 6.0-21 micrograms/m3, while all corresponding background breath concentrations were less than 0.86 micrograms/m3. The internal dose from showering (inhalation plus dermal) was comparable to estimates of the dose from daily water ingestion. The risk associated with a single, 10-min shower was estimated to be 1.22 x 10(-4), while the estimated risk from daily ingestion of tap water ranged from 0.130 x 10(-4) to 1.80 x 10(-4) for 0.15 and 2.0 L, respectively. Since the estimates of chloroform risk from domestic water use for the three exposure routes--ingestion, inhalation, and dermal--are similar, all routes must be used to calculate the total risk when making policy decisions regarding the quality of the municipal water supply.

Air Pollution↗
PubMed2003

Metabolism of chloroform in the human liver and identification of the competent P450s.

The oxidative and reductive cytochrome P450 (P450)-mediated chloroform bioactivation has been investigated in human liver microsomes (HLM), and the role of human P450s have been defined by integrating results from several experimental approaches: cDNA-expressed P450s, selective chemical inhibitors and specific antibodies, correlation studies in a panel of phenotyped HLM. HLM bioactivated CHCl(3) both oxidatively and reductively. Oxidative reaction was characterized by two components, suggesting multiple P450 involvement. The high affinity process was catalyzed by CYP2E1, as clearly indicated by kinetic studies, correlation with chlorzoxazone 6-hydroxylation (r = 0.837; p < 0.001), and inhibition by monoclonal antihuman CYP2E1 and 4-methylpyrazole. The low affinity phase of oxidative metabolism was essentially catalyzed by CYP2A6. This conclusion was supported by the correlation with coumarin 7-hydroxylase (r = 0.777; p < 0.01), inhibition by coumarin and by the specific antibody, in addition to results with heterologously expressed P450s. Chloroform oxidation was poorly dependent on pO(2), whereas the reductive metabolism was highly inhibited by O(2). The production of dichloromethyl radical was significant only at CHCl(3) concentration > or =1 mM, increasing linearly with substrate concentration. CYP2E1 was the primary enzyme involved in the reductive reaction, as univocally indicated by all the different approaches. The reductive pathway seems to be scarcely relevant in the human liver, since it is active only at high substrate concentrations, and in strictly anaerobic conditions. The role of human CYP2E1 in CHCl(3) metabolism at low levels, typical of actual human exposure, provides insight into the molecular basis for eventual difference in susceptibility to chloroform-induced effects due to either genetic, pathophysiological, or environmental factors.

Biotransformation↗
PubMed1975

Genetic control of chloroform toxicity in mice.

Mouse strain differences suggest intermediate or multifactorial gentic control of chloroform-induced renal toxicity and death. The chloroform dose lethal to 50 percent of animals was four times higher in C57BL/6J males than in DBA/2J males. Twice as much chloroform accumulated in the kidneys of the sensitive as the resistant strain. First generation offspring were midway between parental strains for both parameters.

Androgens↗
PubMed1998

Oxidation of trichloroethylene, 1,1-dichloroethylene, and chloroform by toluene/o-xylene monooxygenase from Pseudomonas stutzeri OX1.

Toluene/o-xylene monooxygenase (ToMO) from Pseudomonas stutzeri OX1, which oxidizes toluene and o-xylene, was examined for its ability to degrade the environmental pollutants trichloroethylene (TCE), 1, 1-dichloroethylene (1,1-DCE), cis-1,2-DCE, trans-1,2-DCE, chloroform, dichloromethane, phenol, 2,4-dichlorophenol, 2,4,5-trichlorophenol, 2,4,6-trichlorophenol, 2,3,5,6-tetrachlorophenol, and 2,3,4,5, 6-pentachlorophenol. Escherichia coli JM109 that expressed ToMO from genes on plasmid pBZ1260 under control of the lac promoter degraded TCE (3.3 microM), 1,1-DCE (1.25 microM), and chloroform (6.3 microM) at initial rates of 3.1, 3.6, and 1.6 nmol/(min x mg of protein), respectively. Stoichiometric amounts of chloride release were seen, indicating mineralization (2.6, 1.5, and 2.3 Cl- atoms per molecule of TCE, 1,1-DCE, and chloroform, respectively). Thus, the substrate range of ToMO is extended to include aliphatic chlorinated compounds.

Biodegradation, Environmental↗
PubMed1994

Effects of fecal microorganisms and their chloroform-resistant variants derived from mice, rats, and humans on immunological and physiological characteristics of the intestines of ex-germfree mice.

In order to elucidate the nature of intestinal flora affecting the immunological and physiological parameters of the intestine, we produced several kinds of ex-germfree mice associated with fecal organisms and their chloroform-resistant variants derived from mice, rats, and humans. The phenotypes of intraepithelial lymphocytes were changed to those in conventional mice, particularly the increased positive percentage of alpha beta T-cell-receptor and Thy-1-bearing T cells, on association of the microorganisms (MF) and their chloroform resistant variants (MChl) derived from mice, but not rats and humans, with germfree mice. The cytolytic activity of intraepithelial lymphocytes was expressed only in the MF and MChl groups. The induction of the synthesis of fucosyl asialo GM1 glycolipid, the expression of major histocompatibility complex class II molecule, an increase in the mitotic indices of colonic epithelial cells, and a decrease in lactase activity of the small intestinal epithelial cells also occurred only in the two groups. On the other hand, the cecal size (cecal weight/body weight ratio) was reduced in the mice of all groups examined here, there being approximately the same amount and composition of organic acids, such as acetic acid, butyric acid, and propionic acid, in the cecal contents. Taken together, the results suggest that mouse-specific and chloroform-resistant microorganisms, which are difficult to cultivate at present, may contribute to alteration of the immunological and epithelial characteristics of the mouse intestine. Another factor derived from the intestinal flora, for example, bacterial metabolites such as organic acids, may also affect the cecal size.

Animals↗
PubMed1996

Comparison of hepatitis B virus DNA extractions from serum by the QIAamp blood kit, GeneReleaser, and the phenol-chloroform method.

The abilities of GeneReleaser and QIAamp to extract the hepatitis B virus (HBV) DNA template from serum for amplification by PCR were evaluated and compared with that of the standard phenol-chloroform method. Differences in the sensitivities of the three methods were revealed by nested PCR of HBV DNA extracted from serially diluted hepatitis B e antigen (HBeAg)-positive (high-titer) serum. Phenol-chloroform was found to be the most sensitive extraction method but was time-consuming and labor intensive, and the many steps required increased the possibility of contamination. In a titration of HBeAg-negative (low-titer) serum, all three methods coupled with nested PCR were capable of detecting low levels of HBV DNA. In the case of QIAamp and GeneReleaser, the extraction was relatively simple and rapid. The higher quantity of serum (200 microliters) used in the QIAamp extraction did not provide higher sensitivity, possibly because of incomplete removal of Taq polymerase inhibitors from the serum or inadequate disruption of the virion. GeneReleaser was more efficient because it gave the same detection limit in low-titer serum as phenol-chloroform even though it utilizes only 5 microliters of serum. However, it did not produce consistent amplifications of HBV DNA, giving false-negative results in 7 of the 50 cases (14%) in one experiment. Use of a larger volume of serum and replicate extractions may overcome this problem. Advantages thus exist in each of the extraction methods, and these should be weighed against the disadvantages when deciding which extraction method is appropriate.

Base Sequence↗
PubMed1978

Structure of native and chloroform-methanol-treated mycobacteriophage R1.

The morphologies of native and chloroform-methanol-treated mycobacteriophage R1 were compared by electron microscopy, utilizing three negative stains. R1 was determined to be a complex phage. The head appears as an elongated cylinder with a pointed end (93 +/- 3 by 42 +/- 3 nm) constructed from an orderly arrangement of capsomeres. The phage tail measures 209 +/- 11 by 11 +/- 1 nm and possesses a striated surface with two base plates at its distal end. Treatment of R1 with chloroform-methanol resulted in disruption of both the head and tail structures and was accompanied by loss of infectivity. However, because no likely lipid-containing structure was observed in native phages, there is the possibility that the mechanism of chloroform-methanol inactivation is something other than lipid extraction.

Chloroform↗
PubMed2006

Antinociceptive and anti-inflammatory activities of Dicranopteris linearis leaves chloroform extract in experimental animals.

The present study was carried out to establish the antinociceptive and anti-inflammatory properties of Dicranopteris linearis leaves chloroform extract in experimental animals. The antinociceptive activity was measured using the abdominal constriction, formalin and hot plate tests, while the anti-inflammatory activity was measured using the carrageenan-induced paw edema. The extract, obtained after 72 h soaking of the air-dried leaves in chloroform followed by evaporation under vacuo (40 degrees C) to dryness, was dissolved in dimethyl sulfoxide to the doses of 20, 100 and 200 mg/kg and administered subcutaneously 30 min prior to subjection to the above mentioned assays. The extract, at all doses used, was found to exhibit significant (p<0.05) antinociceptive activity in a dose-dependent manner. However, the significant (p<0.05) anti-inflammatory activity observed occur in a dose-independent manner. As a conclusion, the chloroform extract of D. linearis possesses antinociceptive and anti-inflammatory activity and thus justify its traditional uses by the Malays to treat various ailments.

Animals↗
PubMedDate not supplied

Purification, subunit structure, and kinetics of the chloroform-released F1ATPase complex from Rhodospirillum rubrum and its comparison with F1ATPase forms isolated by other methods.

A stable and homogeneous adenosine-5'-triphosphatase (ATPase, EC 3.6.1.3) has been solubilized from Rhodospirillum rubrum (R. rubrum) chromatophores by chloroform extraction. Purification of the Ca2+-dependent ATPase activity was 200-fold. Ca2+ can be replaced by Mg2+, Cd2+, and Mn2+. The Km for Ca-ATP (0.17 mM) is increased about 5-fold during solubilization of the enzyme, whereas the Km values for Mg-ATP (0.029 mM) and Cd-ATP (0.014 mM) are not affected. The chloroform-released ATPase has a molecular weight of 400,000 +/- 30,000 and consists of the following subunits (molecular weights in parenthesis): alpha(58,000), beta(53,500), gamma(39,000), delta(18,500), and epsilon(14,000). The amino acid composition and the fluorescence spectra are presented. Besides the chloroform-released ATPase complex three other Ca2+-dependent ATPase forms have been isolated from R. rubrum chromatophores by other methods for comparison. Ultrasonication of the membranes leads to the release of an ATPase complex which is mainly composed of alpha, beta, and gamma-subunits. From an acetone powder extract an ATPase complex could be purified by affinity chromatography which is composed of four kinds of subunits (alpha, beta, gamma, delta). The same acetone powder yields an ATPase consisting of only three different types of subunits (alpha, beta, gamma) if the final purification step is preparative disc electrophoresis on 6% polyacrylamide gels instead of affinity chromatography.

Adenosine Triphosphatases↗
PubMed2006

Effectiveness of rotary and hand files in gutta-percha and sealer removal using chloroform or chlorhexidine gel.

The purpose of this study was to assess in vitro the efficacy of nickel-titanium K3 rotary files and hand files for removal of gutta-percha and sealer from obturated root canals using either chloroform or chlorhexidine as solvents. Sixty extracted single-rooted bovine teeth with straight, large canals were prepared, obtured and randomly assigned to 3 groups (n=20). The teeth were stored at 37 degrees C for 1 month and then the gutta-percha and sealer were removed using different techniques, as follows. Group I: size 3 Gates-Glidden drills plus size 30 hand K-files and Hedström files and chloroform; Group II: K3 NiTi rotary files and chloroform; and Group III: K3 NiTi rotary files and 2% chlorhexidine gel. Radiographs were taken and scanned and the images were digitized. The total area of the canal and the area with remaining obturation material were measured in millimeters using a computed image analysis system (ImageLab). Data were analyzed statistically by one-way ANOVA and Tukey test at 5% significance level. The groups differed statistically (p<0.05) with respect to the average percentage of remaining gutta-percha and sealer, presenting the following sequence of effectiveness (from most to least effective): Group I (15.48%), Group II (28.42%) and Group III (35.96%). The findings of this study showed that, despite the technique used for removal of filling material, none of the retreated canals were completely free of gutta-percha and sealer remnants. The use of stainless steel hand files resulted in a lesser amount of filling debris than the use of nickel-titanium rotary instruments.

Animals↗
PubMed1998

Occurrence of toxicity and cell proliferation after a single gavage administration of chloroform to male F344 rats.

Chloroform, an industrial solvent and one of the most common environmental contaminants which produces carcinogenic effects in the liver and kidney of rodents, is not genotoxic in most traditional bacterial and mammalian test systems. Its carcinogenic potential appears attributable to the sustained cell turnover (regenerative hyperplasia) which results from chronic chloroform toxicity. In this present study, cell proliferation (replicative DNA synthesis, RDS) and histopathological changes in hepatocytes and renal tubular epithelial cells were assessed in male F344 rats following a single gavage chloroform exposure (50, 150 or 500 mg/kg). In addition, biochemical parameters (BUN, GOT, LDH and NAG) were examined using plasma and urine samples. Cell proliferation and histopathological changes (e.g. hypertrophy, necrosis, vacuolation) were only seen at the dose of 500 mg/kg in the liver and kidney. At the same dose, all biochemical markers were increased at the 24 to 48 hr time points. These results obtained are thus in line with earlier findings pointing to epigenetic carcinogenicity.

Administration, Oral↗
PubMed1980

The effect of chloroform inhalation on hepatic glucuronidation and de-glucuronidation mechanisms.

The effect of 2, 4, 6 or 8 exposures to chloroform vapour on hepatic glucuronidating (UDPGA transferase) and de-glucuronidating (beta-glucuronidase) levels has been studied in rats. Successive treatments progressively decreased hepatic UDPGA transferase to a minimum of 53% of the control level. beta-Glucuronidase activity was increased two-fold after only two exposures and remained elevated for subsequent exposures. Cytochrome P450 levels decreased with each exposure. The level of this coenzyme in the treated animals remained lower than that of the control animals for at least 48 hours after treatment. UDPGA transferase was diminished to its lowest levels 9 hours after the final exposure to chloroform and did not achieve the control value for a further 48 hours. The beta-glucuronidase activity remained elevated for 12 hours after final exposure. The present experiment demonstrates that inhalation of toxic solvents such as chloroform decreases the glucuronidating capacity of the liver.

Animals↗
PubMed1994

Erythrocyte lipid extraction in alcohol-chloroform systems: a comparative study.

Lipid extraction efficiency is variable when different methods are used on the same tissue. After a systematic comparison on erythrocytes, we found that this variability was due to several factors, such as the addition of alcohol and chloroform as a mixture, the use of repeated extractions or partition procedures after the extraction. Most important, however, was the uneven distribution of solvents and lipids in an apparently monophasic extraction system. This formation reduces lipid extraction and causes accumulation of lipids in the extract supernatant, which leads to an overestimation of the extracted lipid content. We recommend a one-step 18.75 volume of methanol-chloroform (1.5:1 v/v) method for lipid extraction from erythrocytes, with methanol added before chloroform. This method combines simplicity in the extraction procedure, complete extraction of cholesterol and total phospholipids, and reliability in the quantitation of the extracted lipids.

Chloroform↗
PubMedDate not supplied

Some serum biochemical changes associated with chloroform-induced renal tubular necrosis in mice.

The chloroform--induced renal tubular necrosis is associated with the rise of serum urea, creatinine and potassium. The highest values, observed 24 hours after chloroform administration, are respectively: 75 mg% (norm 32 mg%), 1.75 mg% (norm 0.25 mg%), 6.1 mEq/1 (norm 4.4 mEq/1). They gradually decreased with the progress of renal regeneration. These transient, rather little biochemical deviations justify the good conditions and eventual survival of chloroform-poisoned mice. The contribution of possible glomerular lesions to the disturbed renal function is taken into consideration.

Acute Kidney Injury↗
PubMed2002

[Determination of chloroform and tetrachloro-methane in water with capillary column-microECD].

The method of determination of chloroform and tetrachloro-methane in water with capillary column-microECD had better and wider linear range and lower detection limit. At the range of 0-10 micrograms/L chloroform and 0-1 microgram/L tetrachloro-methane, the regression equation were calculated as Y = 1.959X + 4.816 (r = 0.9910) and Y = 3.947X + 42.58 (r = 0.9970). The lowest detection limit were 0.1032 microgram/L chloroform and 0.01219 microgram/L tetrachloro-methane. This method not only simplified the operating processes, but also increase the sensitivity, and reavled better precision and accuracy.

Carbon Tetrachloride↗