ABSTRACT
BACKGROUND/AIMS: Most pesticide formulations contain both chief and additive ingredients. But, the additives may not have been tested as thoroughly as the chief ingredients. The surfactant, nonyl phenoxypolyethoxylethanol (NP40), is an additive frequently present in pesticide formulations. We investigated the effects of NP40 and other constituents of a validamycin pesticide formulation on cell viability and on the expression of genes involved in cell damage pathways. METHODS: The effects of validamycin pesticide ingredients on cell viability and of NP40 on the mRNA expression of 80 genes involved in nine key cellular pathways were examined in the human neuroblastoma SK-N-SH cell line. RESULTS: The chemicals present in the validamycin pesticide formulation were cytotoxic to SK-N-SH cells and NP40 showed the greatest cytotoxicity. A range of gene expression changes were identified, with both up- and down-regulation of genes within the same pathway. However, all genes tested in the necrosis signaling pathway were down-regulated and all genes tested in the cell cycle checkpoint/arrest pathway were up-regulated. The median fold-change in gene expression was significantly higher in the cell cycle checkpoint/arrest pathway than in the hypoxia pathway category (p = 0.0064). The 70 kDa heat shock protein 4 gene, within the heat shock protein/unfolded protein response category, showed the highest individual increase in expression (26.1-fold). CONCLUSIONS: NP40 appeared to be particularly harmful, inducing gene expression changes that indicated genotoxicity, activation of the cell death (necrosis signaling) pathway, and induction of the 70 kDa heat shock protein 4 gene.
Subject(s)
Aged , Female , Humans , Cell Cycle Checkpoints/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Dose-Response Relationship, Drug , Gene Expression Regulation/drug effects , Genes, cdc , HSP110 Heat-Shock Proteins/genetics , Inositol/analogs & derivatives , Necrosis , Neurons/drug effects , Nonoxynol/chemistry , Pesticides/chemistry , RNA, Messenger/metabolism , Signal Transduction/drug effects , Surface-Active Agents/chemistryABSTRACT
BACKGROUND/AIMS: Cyclophosphamide (CP) is a promising treatment for severe cases of paraquat (PQ) poisoning. We investigated the effective dose of CP for mitigating PQ-induced lung injury. METHODS: Adult male Sprague-Dawley rats were allocated into five groups: control, PQ (35 mg/kg, intraperitoneal injection), and PQ + CP (1.5, 15, or 30 mg/kg). The dimensions of lung lesions were determined using X-ray microtomography (micro-CT), and histological changes and cytokine levels were recorded. RESULTS: The micro-CT results showed that 15 mg/kg CP was more effective than 1.5 mg/kg CP for treating PQ-induced lung injury. At a dose of 1.5 mg/kg, CP alleviated the histological evidence of inflammation and altered superoxide dismutase activity. Using 15 mg/kg CP reduced the elevated catalase activity and serum transforming growth factor (TGF)-beta1 level. CONCLUSIONS: A CP dose of > 15 mg/kg is effective for reducing the severity of PQ-induced lung injury as determined by histological and micro-CT tissue examination, possibly by modulating antioxidant enzyme and TGF-beta1 levels.
Subject(s)
Animals , Male , Rats , Catalase/metabolism , Cyclophosphamide/pharmacology , Cytokines/metabolism , Disease Models, Animal , Dose-Response Relationship, Drug , Immunosuppressive Agents/pharmacology , Inflammation Mediators/metabolism , Lung/drug effects , Lung Injury/chemically induced , Oxidative Stress/drug effects , Paraquat , Pulmonary Edema/chemically induced , Rats, Sprague-Dawley , Severity of Illness Index , Superoxide Dismutase/metabolism , Transforming Growth Factor beta1/metabolism , X-Ray MicrotomographyABSTRACT
Chloracetanilide herbicides (alachlor, butachlor, metachlor) are used widely. Although there are much data about chronic low dose exposure to chloracetanilide in humans and animals, there are few data about acute chloracetanilide poisoning in humans. This study investigated the clinical feature of patients following acute oral exposure to chloracetanilide. We retrospectively reviewed the data on the patients who were admitted to two university hospitals from January 2006 to December 2010. Thirty-five patients were enrolled. Among them, 28, 5, and 2 cases of acute alachlor, metachlor, butachlor poisoning were included. The mean age was 49.8 +/- 15.4 yr. The poison severity score (PSS) was 17 (48.6%), 10 (28.6%), 5 (14.3%), 2 (5.7%), and 1 (2.9%) patients with a PSS of 0, 1, 2, 3, and 4, respectively. The age was higher for the symptomatic patients (1-4 PSS) than that for the asymptomatic patients (0 PSS) (43.6 +/- 15.2 vs 55.7 +/- 13.5). The arterial blood HCO3 was lower in the symptomatic patients (1-4 PSS) than that in the asymptomatic patients (0 PSS). Three patients were a comatous. One patient died 24 hr after the exposure. In conclusion, although chloracetanilide poisoning is usually of low toxicity, elder patients with central nervous system symptoms should be closely monitored and cared after oral exposure.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Acetamides/poisoning , Acetanilides/poisoning , Acute Disease , Bicarbonates/blood , Central Nervous System Diseases/diagnosis , Herbicides/poisoning , Poisoning/diagnosis , Retrospective Studies , Severity of Illness Index , Suicide, AttemptedABSTRACT
To identify a prognostic marker that is less sensitive to variations in the elapsed time since paraquat ingestion, we assessed the time between paraquat ingestion and a negative dithionite urine test as a prognostic parameter in patients with acute paraquat intoxication. Forty-one patients with acute paraquat intoxication were enrolled in this study and analyzed to verify significant determinants of mortality and organ dysfunction. The amount of paraquat ingested, paraquat plasma levels, and the time to a negative urine dithionite test were significant independent risk factors predicting mortality. The amount of paraquat ingestion, and the time to a negative urine dithionite test were independent risk factors predicting organ dysfunction. With a cut-off value of 34.5 hr for the time to negative conversion of the urine dithionite test, the sensitivity and specificity for mortality were 71.4% and 75.0%, respectively. The incidence of acute kidney injury and respiratory failure above 34.5 hr were 100% and 85.0%, respectively. In conclusion, the time to a negative urine dithionite test is the reliable marker for predicting mortality and/or essential organ failure in patients with acute paraquat intoxication, who survive 72 hr.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Acute Kidney Injury/etiology , Dithionite/urine , Herbicides/blood , Liver Diseases/etiology , Paraquat/blood , Respiratory Insufficiency/etiology , Risk Factors , Time FactorsABSTRACT
The cellular toxicities of surfactants, a solvent, and an antifreeze that are included in herbicide formulations were assessed by measuring their effects on membrane integrity, metabolic activity, mitochondrial activity, and total protein synthesis rate in a cell culture. Polyethylene glycol, propylene glycol, and monoethylene glycol exhibited no cellular toxicity even at a high concentration of 100 mM. Sodium lauryl ether sulfate and polyoxyethylene lauryl ether significantly damaged the membrane, disturbed cellular metabolic activity, and decreased mitochondrial activity and the protein synthesis rate; however, their toxicity was far below those of the severely toxic chemicals at comparable concentrations. The severely toxic category included polyoxypropylene glycol block copolymer, polyoxyethylene tallow amine, and polyoxyethylene lauryl amine ether. These surfactants were cytotoxic between 3.125 microM and 100 microM in a dose-dependent manner. However, the toxicity graph of concentration vs toxicity had a point of inflection at 25 microM. The slope of the toxicity graph was gentle when the concentration was below 25 microM and steep when the concentration was greater than 25 microM. In conclusion, our results suggest that the toxicity of surfactants be taken care of pertinent treatment of acute herbicide intoxication.
Subject(s)
Animals , Mice , Cell Line , Cell Membrane/drug effects , Herbicides/chemistry , Mitochondria/drug effects , Polyethylene Glycols/toxicity , Sodium Dodecyl Sulfate/toxicity , Surface-Active Agents/chemistry , Toxicity TestsABSTRACT
We investigated whether glyphosate influences the cellular toxicity of the surfactants TN-20 and LN-10 on the mouse fibroblast-like cells, alveolar epithelial cells, and a heart cell line. The cytotoxicity of TN-20 and LN-10 (0.4-100 microM), in the presence or absence of glyphosate was determined by assessing membrane integrity. TN-20 toxicity was significantly lower in the presence of 50 microM glyphosate for the fibroblast-like cell (6.25 microM; 3.9% +/- 3.4% vs -4.8% +/- 0.7%), for the alveolar cells (0.78 microM; 5.7% +/- 0.9% vs 0.1% +/- 0.6%), and for the heart cell line (25.0 microM; 7.9% +/- 3.0% vs 19.4% +/- 0.7%) compared to that of TN-20 alone. The cellular toxicity of LN-10 towards the fibroblast-like cells was found to be increased in the presence of 50 microM glyphosate when LN-10 concentrations of 50 microM (31.3% +/- 3.9% vs 19.2% +/- 0.9%) and 100 microM (62.1% +/- 3.4% vs 39.0% +/- 0.7%) were compared to that of LN-10 alone. These results suggest that the mixture toxicity may be a factor in glyphosate-surfactant toxicity in patients with acute glyphosate herbicide intoxication.
Subject(s)
Animals , Mice , Cell Line , Cell Survival/drug effects , Glycine/analogs & derivatives , Herbicides/chemistry , Polyethylene Glycols/chemistry , Surface-Active Agents/chemistryABSTRACT
BACKGROUND: Culture filtrate proteins secreted by mycobacteria are thought to play an important role in inducing protective immunity and to develop new methods for diagnosing tuberculosis. METHODS: A culture filtrate protein of M. avium that was strongly reactive with goat antiserum against M. intracellulare was constructed. Its homologous protein (TB-14) in M. tuberculosis was cloned, expressed and purified. The inductions of IFN-gamma stimulated with 10 microgram of TB-14 recombinant protein and 10 microgram PPD were estimated by using whole bloods from seven PPD (-) subjects, seven PPD (+) healthy volunteers and nine tuberculosis patients. RESULTS: M. avium culture filtrate protein was confirmed as a hypothetical protein that was termed contig 116. A novel 14-kDa recombinant protein (TB-14) of M. tuberculosis was composed of 148 amino acids, including 30 amino acids of the signal peptide, and it showed 78% homology with M. avium. In the PPD (+) healthy volunteers, recombinant TB-14 protein strongly induced the secretion of IFN-gamma in whole blood cultures. CONCLUSION: These results suggest that TB-14 recombinant protein might play an important role in inducing cell-mediated immunity against tuberculosis. Furthermore, TB-14 protein antigen and its antiserum will be available for the development of new diagnostic tools for tuberculosis.
Subject(s)
Humans , Amino Acids , Clone Cells , Goats , Healthy Volunteers , Immunity, Cellular , Mycobacterium tuberculosis , Mycobacterium , Protein Sorting Signals , TuberculosisABSTRACT
BACKGROUND: The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. METHOD: The PPD positive subjects were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis H37Ra for 4 days by rotating the culture in a 37degrees C, 5% CO2 incubator. In some experiments, methylprednisolone- or pentoxifylline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The delta log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. RESULTS: 1. A trend was noted toward the improved killing of mycobacteria in PPD+subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifylline adversely affected the killing in the PPD+subjects, umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis H37Ra by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis H37Ra. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the delta logKR continuously decreased in a 3 and 4 days of whole blood culture. CONCLUSION: The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.
Subject(s)
Humans , Lung NeoplasmsABSTRACT
Mycobacterium tuberculosis is capable of growing and survival within macrophage. The purpose of this study was to identify the genes regulated by infection of mycobacteria in human monocytic THP-1 cells. We used the differential display reverse transcriptase polymerase chain reaction (DD RT-PCR) and nothern blot analysis to confirm the differentially expressed genes from THP-1 cells infected with live Mycobacterium tuberculosis H37Rv, heat-kille Mycobacterium tuberculosis H37Rv and live Mycobacterium bovis BCG. Among many up or down-regulated clones, 27 clones were sequenced and compared with known genes on GenBank. Thirteen of over-expressed clones from THP-1 cells infected with live Mycobacterium tuberculosis H37Rv were identical to human prothymosin alpha, eight were novel clones and six clones showed homology with Human ferritin H chain, Escherichia coli bgl, Mouse RNA-dependent EIF-2 alpha kinase, E. coli htrL, Hyaluronan receptor and T cell receptor. Our result suggests that Mycobacterium tuberculosis might regulate prothymosin alpha gene transcription in monocytic THP-1 cell.
Subject(s)
Animals , Humans , Mice , Hyaluronan Receptors , Clone Cells , Databases, Nucleic Acid , Escherichia coli , Eukaryotic Initiation Factor-2 , Ferritins , Macrophages , Mycobacterium bovis , Mycobacterium tuberculosis , Mycobacterium , Phosphotransferases , Receptors, Antigen, T-Cell , Reverse Transcriptase Polymerase Chain Reaction , Up-RegulationABSTRACT
PURPOSE: Intravesical instillation of bacillus Calmette-Guerin(BCG) is an established and effective therapy for the superficial bladder carcinoma. The viability of BCG is crucial for the induction of a local immune response as well as effective therapy of recurrent superfical bladder carcinoma. Lubricants are used to facilitate catheterization during intravesical instillation of BCG. Moreover bacteriostatic components contained in them have potential to reduce the viability of the BCG. To verify this assumption, inhibitory effect of four commercially available lubricants on the BCG growth was analyzed. MATERIALS AND METHOD: Four different lubricants and their components were co-incubated with Connaught strain BCG and the resultant growth of BCG was assessed. RESULTS: Significant impairment of BCG viability with lubricants was noted. Chlorhexidine digluconate which is the component of lubricant was considered as responsible for this inhibition. CONCLUSIONS: During intravesical BCG, lubricants might reduce the number of viable BCG in clinical use. For this reason, during intravesical immunotherapy with BCG small amounts of lubricants should be used for urethral catheterization and use of lubricant which does not contain bacteriostatic agent should be considered.