Vikram S. Ghole

Neonatal gram-negative bacteremia

A 22 months prospective study of neonatal gram-negative bacteremia was undertaken in a 15 bed NICU to find out the incidence and antibiotic resistance patterns. Clinically suspected 1326 cases of neonatal sepsis were studied during this period. More than 25% of the cases were microbiologically positive for sepsis. Among 230 (67.2%) cases of gram-negative bacteremia, the predominant isolates werePseudomonas aeruginosa (38.3%),Klebsiella pneumoniae (30.4%),Escherichia coli (15.6%) andAcinetobacter sp. (7.8%). Fifty-nine per cent of the neonates were born in hospital while 41% were from community and referral cases.Lower respiratory tract infection, umbilical sepsis, central intravenous line infection and infection following invasive procedures were the most commonly identified sources of.septicemia. Prematurity and low birth weight were the main underlying conditions in 60% of the neonates. Total mortality was 32%. Increased mortality was mainly associated with rmrhropenia, nosocomial infection and inappropriate antibiotic therapy. Resistance was increasingly noted agains’t many antibiotics. The isolates were predominantly resistant to extended spectrum cephalosporins (25%-75%), piperacillin (68%-78%), and gentamicin (23%-69%).The commonest microorganisms causing gram-negative bacteremia werePseudomonas aeruginosa followed byKlebsiella pneumoniae. The community-acquired bacteremia was mainly due to E.coli. The proportion of preterm and low birth weight babies was significantly high, and the major contributing factor in total mortality. Sensitivity to different antibiotics conclusively proved that a combination of ampicillin + sulbacfam with amikacin or ampicillin + sulbactam with ciprofloxacin is most effective.

In vitro and in vivo genotoxicity assessment of Cr(VI) using comet assay in earthworm coelomocytes

Hexavalent chromium is the major terrestrial pollutant released from electroplating industries, tanneries, thermal power plants and sewage. Cr(VI) is reported genotoxicant in vertebrates. Sub-lethal level of Cr(VI) can cause DNA damage. Earthworms are ecologically important soil invertebrates, used for ecotoxicological studies. In present study coelomocytes of Dichogaster curgensis were used for assessing Cr(VI) genotoxicity using single cell gel electrophoresis [(SCGE) i.e. comet assay] following in vitro and in vivo exposures. Cr(VI) produces reactive oxygen species (ROS) in the cellular system, which can be a major cause of DNA damage. Dose dependent DNA damage was observed during in vitro exposure, while in vivo exposure showed increasing DNA damage till 30ppm in terms of arbitrary units and unexpected decrease in arbitrary units at 70 and 100ppm of Cr(VI). It is speculated that the decrease in arbitrary units at higher concentrations might be due to lower DNA mobility, caused by formation of different types of cross-links.

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.

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.

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.

In vitro genotoxicity of fly ash leachate in earthworm coelomocytes

Fly ash is the major coal combustion byproduct from thermal power plants. Considering its plant–nourishing constituents, its soil amendment in farmland is one of its promoted disposal methods. A substantial amount of heavy metals present in fly ash, which may leach out due to rainwater or irrigation water, may cause serious problem with long term use, especially to soil organisms. These metals may cause DNA damage through Reactive Oxygen Species (ROS) generation. In the present study, single cell gel electrophoresis [(SCGE) i.e., comet assay] was used to detect DNA damage in earthworm (Dichogaster curgensis) coelomocytes, following an in vitro exposure. Significant DNA damage was observed at the lowest concentration of fly ash leachate (6.25%) examined. DNA damage by all the tested concentrations (6.25%, 12.5%, 25%, 50%) differed significantly (p < 0.001) from that of the negative control. Hence, long-term application of fly ash might prove harmful for earthworm populations.

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. Fishers 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.

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.

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.

PER-1-type extended-spectrum β-lactamase-producing Acinetobacter baumannii clinical isolates from India

c t m p i E r e o s f n C g f H Fig. 1. Demonstration of extended-spectrum -lactamase (ESBL) from Acinetobacter baumannii isolates. (A) Detection by double-disk synergy test (DDST) and phenotypic confirmatory disk diffusion test (PCDDT). Ac, amoxicillin/clavulanic acid; Ca, ceftazidime; Ce, cefotaxime; Ci, ceftriaxone; CPM, cefepime; Ao, aztreonam; FOX, 88 Letters to the Editor / International Journ