Proteomics study revealed altered proteome of Dichogaster curgensis upon exposure to fly ash.

Fly ash is toxic and its escalating use as a soil amendment and disposal by dumping into environment is receiving alarming attention due to its impact on environment. Proteomics technology is being used for environmental studies since proteins respond rapidly when an organism is exposed to a toxicant, and hence soil engineers such as earthworms are used as model organisms to assess the toxic effects of soil toxicants. This study adopted proteomics technology and profiled proteome of earthworm Dichogaster curgensis that was exposed to fly ash, with main aim to elucidate fly ash effects on cellular and metabolic pathways. The functional classification of identified proteins revealed carbohydrate metabolism (14.36%), genetic information processing (15.02%), folding, sorting and degradation (10.83%), replication and repair (3.95%); environmental information processing (2.19%), signal transduction (9.61%), transport and catabolism (17.27%), energy metabolism (6.69%), etc. in the proteome. Proteomics data and functional assays revealed that the exposure of earthworm to fly ash induced protein synthesis, up-regulation of gluconeogenesis, disturbed energy metabolism, oxidative and cellular stress, and mis-folding of proteins. The regulation of ubiquitination, proteasome and modified alkaline comet assay in earthworm coelomocytes suggested DNA-protein cross link affecting chromatin remodeling and protein folding.

Biomarker responses in the earthworm, Dichogaster curgensis exposed to fly ash polluted soils.

Earthworms are globally accepted as a model organism in terrestrial ecotoxicology for assessment of environmental pollution. This study evaluated and compared effects of fly ash polluted soils collected from two geographically different thermal power plants on biomarker responses in the earthworm, Dichogaster curgensis. To evaluate relationship between distance sampling and biomarker responses in the earthworm D. curgensis, soil samples at 0.5, 1 and 3km from thermal plant were analyzed for physico-chemical properties and metal concentrations. Biochemical alterations, lysosomal membrane stability, genotoxic effects, and histological changes were examined on 1, 7, and 14 d of exposure to fly ash contaminated soils collected from different thermal power plants. The activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) levels were significantly increased, while glutathione reductase (GR) activity was found to be decreased in treated animals. Catalase (CAT) and glutathione-S- transferase (GST) activities were found to be increased initially up to 7d exposure and further decreased on 14d exposure. D. curgensis exposed to fly ash contaminated soils showed significant lysosomal membrane destabilization and DNA damage. Extensive histopathological changes were observed in the tissues of the body wall and intestinal tract of the exposed D. curgensis along with accumulation of heavy metals. These results demonstrate that soil pollution around thermal power plants has adverse biological effects of on the indicator organism D. curgensis and no correlation was found between distance and extent of biological biochemical responses.

Chemical Treatment as a Pre- requisite for Effective Disposal of Tri Nitro Toluene (TNT).

Present methods for the disposal of old and rejected ammunition carry high risk and are not environment-friendly. Various processes such as wet air oxidation, molten salt oxidation, hydro thermal oxidation, incineration, electrochemical reduction, biodegradation and other methods have limited use for decontamination and are not suitable for disposal of large quantities of explosives. Thus there is dire need to develop alternate method for safe disposal of rejected explosives which will be eco-friendly. In this paper we have attempted to combine two methods i.e. chemical treatment followed by biological/microbiological treatment. For this purpose we have selected Tri Nitro Toluene (TNT) as a model compound, which is used extensively in many types of ammunition. As reported previously from our laboratory the presence of nitro group from TNT was toxic to bacterial growth. By chemical treatment, nitro groups from TNT were converted into amine and mixed in soil for biodegradation. Our results suggest that after converting 'nitro groups' to 'amine groups' were much preferred by bacteria and faster mineralization is observed. Thus combined treatment to TNT as discussed in this study, showed much less phyto-toxicity and may have great potential to scale up the process for large quantities of explosive such as TNT.

Proteomic profiling and interactome analysis of ER-positive/HER2/neu negative invasive ductal carcinoma of the breast: towards proteomics biomarkers.

Breast cancer, especially ER positive/HER2/neu negative IDC, is the predominant subtype of invasive ductal carcinoma. Although proteomic approaches have been used towards biomarker discovery in clinical breast cancer, ER positive/HER2/neu negative IDC is the least studied subtype. To discover biomarkers, as well as to understand the molecular events associated with disease progression of estrogen receptor positive/HER2/neu negative subtype of invasive ductal carcinoma, differential protein expression profiling was performed by using LC-MS(E) (MS at elevated energy). A total of 118 proteins were identified, of which 26 were differentially expressed. These identified proteins were functionally classified and their interactions and coexpression were analyzed by using bioinformatic tools PANTHER (Protein Analysis THrough Evolutionary Relationships) and STRING (Search Tool for the Retrieval of Interacting Genes). These proteins were found to be upregulated and were involved in cytoskeletal organization, calcium binding, and stress response. Interactions of annexin A5, actin, S100 A10, glyceraldehyde 3 phosphate dehydrogenase, superoxide dismutase 1, apolipoprotein, fibrinogen, and heat shock proteins were prominent. Differential expression of these proteins was validated by two-dimensional gel electrophoresis and Western blot analysis. The cluster of these proteins may serve as a signature profile for estrogen receptor positive/ HER2/neu negative subtype.

Effect of fly ash on biochemical responses and DNA damage in earthworm, Dichogaster curgensis.

Fly ash is receiving alarming attention due to its hazardous nature, widespread usage, and the manner of disposal; leading to environmental deterioration. We carried out bio-monitoring and risk assessment of fly ash in earthworms as a model system. Dichogaster curgensis were allowed to grow in presence or absence of fly ash (0-40%, w/w) for 1, 7, and 14 d. The biochemical markers viz. catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), glutathione peroxidase (GPx), glutathione S-transferase (GST), and malondialdehyde (MDA) level were measured. The comet and neutral red retention assays were performed on earthworm coelomocytes to assess genetic damages and lysosomal membrane stability. The results revealed increased activities of SOD, GPx, GST, and MDA level in a dose-response manner while GR activity was decreased with increasing concentrations of fly ash. No obvious trend was observed in the CAT activity and fly ash concentration. Lysosomal membrane destabilization was noted in the earthworms exposed to 5% and more fly ash concentration in a dose and time dependent manner. The comet assay demonstrated that the fly ash induced DNA damage and DNA-protein crosslinks in earthworm coelomocytes.

Analysis of AGE modified proteins and RAGE expression in HER2/neu negative invasive ductal carcinoma.

Cancer is associated with increased glycolysis and carbonyl stress. In view of this, AGE modified proteins were identified from clinical breast cancer tissue using 2DE-immunoblot and mass-spectrometry. These proteins were identified to be serotransferrin, fibrinogen gamma chain, glycerol-3-phosphate dehydrogenase, lactate dehydrogenase, annexin II, prohibitin and peroxiredoxin 6, which have established role in cancer. Further, RAGE expression and its downstream signaling proteins NADPH oxidase and NF-kB were studied. Role of these AGE modified proteins and RAGE signaling in breast cancer is discussed.

Effect of a novel biphenyl compound, VMNS2e on ob/ob mice.

VMNS2e is a novel biphenyl compound, which in previous studies had showed most favourable interactions with the active site of protein tyrosine phosphatase 1B (PTP1B). The effect of acute and chronic treatment of VMNS2e (30mg/kg) was investigated in ob/ob mice. Plasma glucose was measured after acute administration of VMNS2e (30mg/kg) in both lean and ob/ob mice. In the chronic study, VMNS2e (30mg/kg) was given orally, once daily for 60days. Metformin (300mg/kg) was taken as standard therapy. Body weight, food intake and blood glucose was measured weekly while glycosylated hemoglobin A(1c) (HbA(1c)), insulin, triglyceride, total cholesterol, low density lipoprotein (LDL), fructosamine, non esterified fatty acid and organ weight were estimated after the completion of treatment period. Oral glucose tolerance test was performed on the last day of treatment. Liver and epididymal fat weights were taken. Acute dose of VMNS2e elicited an anti hyperglycemic effect. It reduced blood glucose by 14% (0.5h) and 35.6% (6h). Chronic VMNS2e treatment improved glucose tolerance by 25.3%. It decreased blood glucose levels. Hyperinsulinemia was reduced (19.6%). VMNS2e treatment had no significant effect on body weight and food consumption. VMNS2e treatment exhibited significant reduction (28.2%) in HbA(1c), plasma triglyceride (49%), LDL (24%) and fructosamine (13%) levels. VMNS2e treatment did not alter total cholesterol and non esterified fatty acid levels. Epididymal fat/body weight ratio was reduced (26.3%). VMNS2e exhibited both acute and chronic anti hyperglycemic effect, insulin sensitivity along with improvement in various lipid parameters and glycemic control.

Novel biphenyl compound, VMNS2e, ameliorates streptozotocin-induced diabetic nephropathy in rats.

AIM: To study the effect of a new biphenyl synthetic compound showing interactions with the active site of protein tyrosine phosphatase 1B by docking and molecular dynamics, VMNS2e in streptozotocin-induced diabetic nephropathy in rats with various renal function parameters and renal ultrastructure. METHODS: Streptozotocin (55 mg/kg)-induced diabetic rats were orally treated once daily with VMNS2e (30, 60, and 120 mg/kg) for 8 weeks. The body weight and blood glucose levels of the rats were recorded during the study period. After 8 weeks of treatment creatinine clearance, urinary protein, blood urea nitrogen, urinary albumin excretion rate, and insulin levels were measured. An ultrastructure study of the kidney tissue was performed and the glomerular basement membrane thickness was measured. RESULTS: Eight weeks of VMNS2e treatment significantly reduced the fasting blood glucose level, attenuated elevating blood urea nitrogen levels, and reduced glomerular basement membrane thickness. CONCLUSION: It is concluded that VMNS2e treatment at 30 and 60 mg/kg, when given for 8 weeks, partly ameliorated early diabetic nephropathy in diabetic rats.

Assessment of air pollution and its effects on the health status of the workers in beam rolling mills factory (Iran National Steel Industrial Group) from Ahvaz-Iran.

BACKGROUND: Air pollutants of iron- and steel-making operations have historically been an environmental and health hazard. These pollutants include gaseous substances such as sulfur oxide, nitrogen dioxide, and carbon monoxide. The Iran National Steel Industrial Group beam rolling mills factory has two production lines viz. line 630 and line 650, with different beam production capabilities and is capable of producing different types of beams. MATERIALS AND METHODS: A retrospective cross-sectional study on 400 workers in different exposure levels to environmental pollution was performed during 2005 to determine the mean value of respirable particulate matter (RPM) concentrations and its effects on the health status of workers. To elicit information regarding the health status of the worker, the National Institute for Occupational Safety and Health standard questionnaire was used. Fisher's exact test was performed to assess the relative risk (RR) of exposure to air pollution on cardiovascular diseases, chest tightness, cough, difficulty in retention, i.e. loss of memory, tension, occupational fatigue, and occupational stress in exposed workers. RESULTS: There was significant difference in RPM pollution level between two product lines. The RR of exposure to air pollution on cardiovascular diseases, chest tightness, cough, difficulty in retention, i.e. loss of memory, tension, occupational fatigue, and occupational stress in exposed workers were 2.78, 2.44, 2.15, 1.92, 1.57, 3.90, and 2.09, respectively.

Aerobic reduction of perchlorate by bacteria isolated in Kerala, South India.

Possibility of perchlorate reduction by microbes raises hope for an eco-friendly mode of degradation of this toxic rocket fuel. This study reports 3 isolates (A1, A2 and A3) capable of molybdenum-independent degradation of perchlorate under aerobic conditions. The rate of degradation was the highest when perchlorate concentration was 17 mM, and then 3.2 mM, 4.7 mM and 4.1 mM of perchlorate was reduced by isolates A1, A2 and A3 (respectively) after 72 h at 28 degrees C under aerobic conditions. Presence of perchlorate at a concentration higher than 17 mM resulted in some inhibition of perchlorate reduction. 16S ribosomal RNA gene analysis revealed isolate A1 to be Pseudomonas stutzeri (Proteobacteria) while isolates A2 ad A3 where found to belong to the genus Arthrobacter (Actinobacteria). The study, apart from demonstrating ribotyping as a rapid method of identification of economically important soil microbes, also raised prospects for using artificial consortia for environmental degradation of perchlorate, without apparent domination of Dechloromonas spp. (a group of microbes known for perchlorate remediation in the environment).

Investigation of respirable particulate matter pollutants on air-breathing zone workers in the Beam Rolling Mills Factory (Iran National Steel Industrial Group), Ahvaz, Iran.

Workers of iron and steel factories are exposed to a wide range of pollutants depending on the particular process, the materials involved, the effectiveness of monitoring and the control measures. Adverse effects are determined by the physical state and propensities of the pollutant involved, the intensity and duration of the exposure, the extent of pollutant accumulation in the body and the sensitivity of the individual to its effects. The main aim of this study is to assess the levels of the indoor respirable particulate matter (RPM) and to compare the health condition of exposed workers, with nonexposed employees group. Line 630 has only one furnace of 40 tons and line 650 has two furnaces of 20 and 40 tons capacity due to which the mean of the RPM concentrations in the breathing zone was significantly different (P < 0.05) in line 650 but not in line 630 as compared with National Institute for Occupational Safety and Hygiene's (3 mg/m(3)). The average of the RPM concentrations in production line 650 is higher than that of production line 630, with the 95% confidence interval in saw cabin station number 1 of production line 650.

Clinical and demographic features of infection caused by Acinetobacter species.

BACKGROUND: Recently, Acinetobacter emerged as an important pathogen and the prevalence of isolation has increased since the last two decades worldwide. AIMS: To determine Acinetobacter incidence, their clinical demography, antibiotyping and speciation. SETTINGS AND DESIGN: A study of the clinical samples submitted to microbiology laboratory of a teaching hospital over a period of 3 years (December 1994 through November 1997). MATERIALS AND METHODS: Identification, speciation and antibiotyping were performed for the isolates of Acinetobacter recovered from infective samples. Clinical demographic characteristics were studied retrospectively. RESULTS: Total 510 of 5391 (9.6%) of isolates were Acinetobacter, responsible for 71.2% (363 of 510) monomicrobial and 28.8% (147 of 510) polymicrobial infections. The organism was responsible for 156 (30.6%) cases of urinary tract infection and 140 (27.5%) cases of wound infection and was most prevalent in the intensive care unit (30.8%, 140 of 455). The crude mortality rate due to multi-drug resistant Acinetobacter septicemia was 7.9% (36 of 455). The isolates could be classified into 7 species, with A. baumannii being most predominant. No peculiar pattern during antibiotyping was observed, but most of them were multi-drug resistant. CONCLUSION: Multi-drug resistant Acinetobacter nosocomial infection has emerged as an increasing problem in intensive care units of the hospital, responsible for 7.9% deaths. The analysis of risk factors and susceptibility pattern will be useful in understanding epidemiology of this organism in a hospital setup.

Alcohol dehydrogenase: a potential new marker for diagnosis of intestinal ischemia using rat as a model.

AIM: Intestinal ischemia (Ii) is an abdominal emergency due to blockade of the superior mesenteric artery resulting in 60-100% mortality if diagnosed late. Changes in several biochemical parameters such as D (-)-lactate, Creatinine kinase isoenzymes and lactate dehydrogenase suggested for early diagnosis, lack specificity and sensitivity. Therefore a biochemical parameter with greater sensitivity needs to be identified. METHODS: Wistar male rats were randomly assigned into two groups; control sham operated (n = 24) and ischemic test (n = 24) group. Superior mesenteric arterial occlusion was performed in the ischemic test group for 1 h. Alcohol dehydrogenase (ADH) was estimated in blood from portal vein, right ventricle of heart, dorsal aorta (DA) and inferior vena cava (IVC). The Serum glutamic acid pyruvate transaminase (SGPT) was also estimated in blood from portal vein and right ventricle of heart. RESULTS: A significant increase (P<0.001) in the levels of ADH in both portal blood as well as heart blood of the test group (232.72+/-99.45 EU and 250.85+/-95.14 EU, respectively) as compared to the control group (46.39+/-21.69 EU and 65.38+/-30.55 EU, respectively) were observed. Similarly, increased levels of ADH were observed in blood samples withdrawn from DA and IVC in test animals (319.52+/-80.14 EU and 363.90+/-120.68 EU, respectively) as compared to the control group (67.68+/-63.22 EU and 72.50+/-58.45 EU, respectively). However, in test animals there was significant increase in SGPT in portal blood (P = 0.054) without much increase in heart blood. CONCLUSION: Significant increase in the levels of ADH in portal and heart blood within 1 h of SMA occlusion without increase in SGPT in heart blood, suggests that the origin of ADH is from ischemic intestine and not from liver. Similarly, raised ADH levels were found in DA and IVC as well. IVC blood does represent peripheral blood sample. A raised level of ADH in test animals confirms it to be a potential marker in the early diagnosis of Ii.

Multidrug resistant Acinetobacter baumannii isolates from a teaching hospital.

Abstract Acinetobacter baumannii is the most common nosocomial pathogen among all Acinetobacter spp. A. baumannii tend to be resistant to multiple antibiotics, and represent a severe threat in the treatment of hospitalized patients. The purpose of this study was to evaluate the prevalence, biotyping, and antibiotic resistance status of A. baumannii isolates recovered from submitted hospital clinical specimens. The in-vitro activity of 29 currently used antimicrobial agents was studied in 180 isolates of A. baumannii. The prevalence was higher in wound exudates (32%) and urine samples (22%) than in other specimens. Forty-six percent of the isolates were of biotype 9. The evolution of resistance from November 1996 through October 1998 was studied. More than 75% of the isolates were multidrug resistant (MDR) and more than 70% were #-lactamase producers. Amikacin, ampicillin + sulbactam, and imipenem are still effective antimicrobial agents, but a steady rise in the values of the minimum inhibitory concentration at which 50% of the isolates were inhibited (MIC50 )and MIC90 was observed. Resistance to aztreonam (60%), enrofloxacin (60%), imipenem (29%), moxalactam (73%), and sparfloxacin (25%) was noted; enrofloxacin and sparfloxacin are not generally used in this hospital. From the findings of the present study, no single drug appears to be suitable for empirical therapy for this nosocomial pathogen. Abstract Acinetobacter baumannii is the most common nosocomial pathogen among all Acinetobacter spp. A. baumannii tend to be resistant to multiple antibiotics, and represent a severe threat in the treatment of hospitalized patients. The purpose of this study was to evaluate the prevalence, biotyping, and antibiotic resistance status of A. baumannii isolates recovered from submitted hospital clinical specimens. The in-vitro activity of 29 currently used antimicrobial agents was studied in 180 isolates of A. baumannii. The prevalence was higher in wound exudates (32%) and urine samples (22%) than in other specimens. Forty-six percent of the isolates were of biotype 9. The evolution of resistance from November 1996 through October 1998 was studied. More than 75% of the isolates were multidrug resistant (MDR) and more than 70% were #-lactamase producers. Amikacin, ampicillin + sulbactam, and imipenem are still effective antimicrobial agents, but a steady rise in the values of the minimum inhibitory concentration at which 50% of the isolates were inhibited (MIC50) and MIC90 was observed. Resistance to aztreonam (60%), enrofloxacin (60%), imipenem (29%), moxalactam (73%), and sparfloxacin (25%) was noted; enrofloxacin and sparfloxacin are not generally used in this hospital. From the findings of the present study, no single drug appears to be suitable for empirical therapy for this nosocomial pathogen.