Architectural analysis of root system and phytohormone biosynthetic genes expression in wheat (Triticum aestivum L.) inoculated with Penicillium oxalicum.

In this study, a fungal plant growth promoter Penicillium oxalicum T4 isolated from non-rhizosphere soil of Arunachal Pradesh, India, was screened for different plant growth promoting traits in a gnotobiotic study. Though inoculation improved the overall growth of the plants, critical differences were observed in root architecture. Confocal Laser Scanning Microscope, Scanning electron microscope and the stereo microscopic study showed that inoculated wheat plants could develop profuse root hairs as compared to control. Root scanning indicated improvement in cumulative root length, root area, root volume, number of forks, links, crossings, and other parameters. A confocal scanning laser microscope indicated signs of endophytic colonization in wheat roots. Gene expression studies revealed that inoculation of T4 modulated the genes affecting root hair development. Significant differences were marked in the expression levels of TaRSL4, TaEXPB1, TaEXPB23, PIN-FORMED protein, kaurene oxidase, lipoxygenase, ACC synthase, ACC oxidase, 9-cis-epoxycarotenoid dioxygenase, and ABA 8'-hydroxylase genes. These genes contribute to early plant development and ultimately to biomass accumulation and yield. The results suggested that P. oxalicum T4 has potential for growth promotion in wheat and perhaps also in other cereals.

Maternal Spirulina supplementation during pregnancy and lactation partially prevents oxidative stress, glial activation and neuronal damage in protein malnourished F1 progeny.

Protein malnutrition (PMN) is a global health issue but most prevalent in Africa and Asia. It exerts detrimental effect on structural and physiological aspects of hippocampal circuitry. Despite accumulating evidence for PMN induced changes in nervous system, relatively very little is known about how maternal nutritional supplementation during malnutrition affects glial cells and neurons. Herein, we aimed to investigate the effects of maternal Spirulina supplementation against PMN induced oxidative stress, reactive gliosis and neuronal damage in hippocampus of F1 progeny. Three months old healthy Sprague Dawley females (n = 24) were shifted to normoprotein (NC; 20% protein) and low protein (LP; 8% protein) diets 15 days before conception. The NC and LP group females were subdivided into two groups according to Spirulina supplementation (400 mg/kg/b.wt. orally throughout gestation and lactation period): normal control with Spirulina (NC SPI) and low protein with Spirulina supplemented group (LP SPI). F1 progeny born were used in present study. Thus, building on earlier results of ameliorated neurobehavioral and cognitive abilities in Spirulina supplemented protein deprived rats, the present study incorporates neurochemical and morphometric analysis of glial cells and neurons and revealed that maternal Spirulina consumption partially prevented the PMN associated neuropathological alterations in terms of attenuated oxidative brain damage, reduced reactive gliosis and apoptotic cell population, improved dendritic branch complexity with few damaged neurons and enhanced mushroom shaped spine density. The results suggest that cellular changes in hippocampus after PMN are partially restored after maternal Spirulina supplementation and one could envision intervention approaches using Spirulina against malnutrition.

Rapid analysis of pharmaceutical drugs using LIBS coupled with multivariate analysis.

Type 2 diabetes drug tablets containing voglibose having dose strengths of 0.2 and 0.3 mg of various brands have been examined, using laser-induced breakdown spectroscopy (LIBS) technique. The statistical methods such as the principal component analysis (PCA) and the partial least square regression analysis (PLSR) have been employed on LIBS spectral data for classifying and developing the calibration models of drug samples. We have developed the ratio-based calibration model applying PLSR in which relative spectral intensity ratios H/C, H/N and O/N are used. Further, the developed model has been employed to predict the relative concentration of element in unknown drug samples. The experiment has been performed in air and argon atmosphere, respectively, and the obtained results have been compared. The present model provides rapid spectroscopic method for drug analysis with high statistical significance for online control and measurement process in a wide variety of pharmaceutical industrial applications.

The role of algae in agriculture: a mathematical study.

Synthetic fertilizers and livestock manure are nowadays widely used in agriculture to improve crop yield but nitrogen and phosphorous runoff resulting from their use compromises water quality and contributes to eutrophication phenomena in waterbeds within the countryside and ultimately in the ocean. Alternatively, algae could play an important role in agriculture where they can be used as biofertilizers and soil stabilizers. To examine the possible reuse of the detritus generated by dead algae as fertilizer for crops, we develop three mathematical models building upon each other. A system is proposed in which algae recover waste nutrients (nitrogen and phosphorus) for reuse in agricultural production. The results of our study show that in so doing, the crop yield may be increased and simultaneously the density of algae in the lake may be reduced. This could be a way to mitigate and possibly solve the environmental and economic issues nowadays facing agriculture.

Modeling the impact of awareness on the mitigation of algal bloom in a lake.

The proliferation of algal bloom in water bodies due to the enhanced concentration of nutrient inflow is becoming a global issue. A prime reason behind this aquatic catastrophe is agricultural runoff, which carries a large amount of nutrients that make the lakes more fertile and cause algal blooms. The only solution to this problem is curtailing the nutrient loading through agricultural runoff. This could be achieved by raising awareness among farmers to minimize the use of fertilizers in their farms. In view of this, in this paper, we propose a mathematical model to study the effect of awareness among the farmers of the mitigation of algal bloom in a lake. The growth rate of awareness among the farmers is assumed to be proportional to the density of algae in the lake. It is further assumed that the presence of awareness among the farmers reduces the inflow rate of nutrients through agricultural runoff and helps to remove the detritus by cleaning the bottom of the lake. The results evoke that raising awareness among farmers may be a plausible factor for the mitigation of algal bloom in the lake. Numerical simulations identify the most critical parameters that influence the blooms and provide indications to possibly mitigate it.

Molecular characterization and expression analysis of hsp60 gene homologue of sheep blowfly, Lucilia cuprina.

The 60kDa heat shock protein (Hsp60) or chaperonin is one among the highly conserved families of heat shock proteins, known to be involved in variety of cellular activities, including protein folding, thermal protection, etc. In this study we sequence characterized hsp60 gene homologue of Lucilia cuprina, isolated and cloned from the genomic library as well as by genomic PCR, followed by RACE- PCR. The L. cuprina hsp60 gene/protein expression pattern was analyzed in various tissues, either at normal temperature (25±1°C) or after exposure to heat stress (42°C). The analysis of nucleotide sequence of Lchsp60 gene revealed absence of intron and the nuclear localizing signal (NLS). The deduced amino acid sequence showed presence of unique conserved sequences, such as those for mitochondrial localization, ATP binding, etc. Unlike Drosophila, Lucilia showed presence of only one isoform, i.e., hsp60A. Phylogenetic analysis of hsp60 gene homologues from different species revealed Lchsp60 to have >88.36% homology with D. melanogaster, 76.86% with L. sericata, 58.31% with mice, 57.99% with rat, and 57.72% with human. Expression analysis using Real Time PCR and fluorescence imaging showed significant enhancement in the expression level of Lchsp60 upon heat stress in a tissue specific manner, indicating its likely role in thermo-tolerance as well as in normal cellular activities.

Spatial dynamics of a nutrient-phytoplankton system with toxic effect on phytoplankton.

The production of toxins by some species of phytoplankton is known to have several economic, ecological, and human health impacts. However, the role of toxins on the spatial distribution of phytoplankton is not well understood. In the present study, the spatial dynamics of a nutrient-phytoplankton system with toxic effect on phytoplankton is investigated. We analyze the linear stability of the system and obtain the condition for Turing instability. In the presence of toxic effect, we find that the distribution of nutrient and phytoplankton becomes inhomogeneous in space and results in different patterns, like stripes, spots, and the mixture of them depending on the toxicity level. We also observe that the distribution of nutrient and phytoplankton shows spatiotemporal oscillation for certain toxicity level.

Gallbladder cancer incidence in Gwalior district of India: Five-year trend based on the registry of a regional cancer center.

BACKGROUND: We have reported here the 5-year incidence (2004-2008) of gallbladder cancer (GBC) in North Central India along with its descriptive epidemiology. This provides potential clues for better prevention. The present study has also evaluated the association of ABO blood groups with GBC. PATIENTS AND METHODS: The study comprised 742 GBC cases referred to the regional cancer hospital, Gwalior, during 2004-2008. The demographic statistics of Gwalior district was considered to calculate the relative risk and incidence rates. ABO blood group distribution amongst 90,000 healthy subjects registered in the local blood bank during 2002-2007 was taken as controls to study the association of blood groups with GBC. RESULTS: The age-standardized total incidence rate of GBC was calculated to be 7.16/1,00,000. The relative risk of females getting GBC was 2.693 at 95% confidence interval of 2.304-3.151 (P < 0.0001). The females formed 69.5% of total cancer cases, with age-standardized incidence rate of 10/1,00,000. The mean age of male and female GBC cases was found to be 55.4 years (SD = 13, SE = 0.77) and 51.5 years (SD = 12.3, SE = 0.50), respectively. The blood groups A (P = 0.0022) and AB (P < 0.0001) had a positive association with GBC with significant level of differences in comparison to controls. CONCLUSION: Our study provided an estimate of a 5-year incidence of GBC in North Central India for the first time. With regard to the association of risk factors like obesity, age, and urban living with GBC, the findings of the present study are contradictory to the general opinion. Blood groups A and AB were found to be associated with GBC, which would be provisional for further investigations.

UGT1A1 gene variants and clinical risk factors modulate hyperbilirubinemia risk in newborns.

OBJECTIVE: To study the contribution of UGT1A1 gene variants and clinical risk factors in modulating hyperbilirubinemia risk in newborns. STUDY DESIGN: Seven UGT1A1 gene variants and clinical risk factors were studied in 113 hyperbilirubinemia cases and 218 control newborns. Hyperbilirubinemia was defined as the total serum bilirubin levels >95th percentile of the American Academy of Pediatrics nomogram. The study population included term (37 to 41 weeks) newborns below 2 weeks of age. RESULT: UGT1A1 gene variants, namely, c.211G>A, g.-3279T>G, TATA box polymorphism and CAT insertion were identified as independent molecular risk factors for neonatal hyperbilirubinemia, whereas c.686C>A, c.1091C>T and c.1456T>G were not detected in study cohort. Among clinical risk factors, excessive weight loss, sepsis and ABO incompatibility emerged as independent risk factors. Co-expression of UGT1A1 variants and clinical risk factors further accentuated the risk of neonatal hyperbilirubinemia. CONCLUSION: Multiple risk factors, whether genetic or clinical, are instrumental in modulating hyperbilirubinemia risk in newborns. Disordered bilirubin conjugation through interactions of UG1TA1 gene variants contributes to the clinical phenotype of neonatal hyperbilirubinemia.

Value of image-guided fine-needle aspiration cytology-a study of 500 cases.

Guided Fine-Needle Aspiration Cytology (FNAC) as the first line investigation is not only useful in diagnosis of space occupying lesions but can also help in choosing appropriate management. This technique is most useful in diagnosing metastasis but is also helpful in excluding malignancy in some cases. Aim of this study is to analyze the spectrum of cytological diagnosis, adequacy, and clinicoradiological correlation of guided FNAC. The study was carried out in the departments of pathology and radiology for a period from January 2000 to November 2011. All the data were retrieved from hospital medical record section and departmental data bank and were analyzed. We have performed a total of 500 FNACs, including 234 under CT guidance and 266 under USG guidance. Among the CT-guided FNACs, 87.6% (205 cases) had adequate material which included 152 malignant cases, 38 inflammatory cases and 15 benign cases. Among the cases under USG, 91.7% (244 cases) had adequate material comprising of 112 malignant cases, 44 inflammatory cases, and 88 benign lesions. It was noted that increased number of inadequate aspirate was from lesions with smaller size and increased depth. Out of total 264 malignant cases diagnosed on FNAC, 160 cases were identified as malignant on clinical and radiological findings. Guided FNAC is useful in deep seated lesions as well as in suspicious superficial lesions and is a safe diagnostic tool. If performed properly and correlated well with clinical and radiological findings, FNAC provides correct diagnosis in most cases.

The liver function test enzymes and glucose level are positively correlated in gallbladder cancer: a cancer registry data analysis from north central India.

AIM OF STUDY: To investigate the trend of expression of liver function test enzymes and other biochemical changes during gallbladder carcinogenesis. MATERIALS AND METHODS: Eight hundred and seventy-eight gallbladder disease patients were selected to study the liver function test enzymes and routine blood biochemical changes in the last five years (2004-08). Statistical analysis was performed using Graph Pad prism 5.02 software. RESULTS: The liver function test enzymes showed significant correlations among themselves, and with glucose in gallbladder cancer and gallstone disease patients (N = 878). Out of 878 gallbladder cases, 46 (5.24%) showed significantly higher glucose level of 216.66 mg/dL (P < 0.0001). All the three pathological conditions of gallbladder, gallbladder cancer with stones (GBCS), gallbladder cancer without stones (GBC) and calculus cholecystitis (CC), showed highly significant positive correlation (Pearson) between Serum Glutamic Oxaloactetic Transaminase (SGOT) and Serum Glutamic Pyruvic Transaminase (SGPT) [P < 0.0001, (GBCS); P < 0.0001, (GBC), and P < 0.0001, (CC)]. SGOT and SGPT also showed positive correlation with higher glucose level independently, in both GBCS and CC (P < 0.0001 and P < 0.0001), respectively. CONCLUSION: Simultaneous elevation of glucose and liver function test enzymes in GBC makes the diagnosis complex. Any patient of gallbladder diseases with higher level of glucose may have the possibility of developing gallbladder cancer.

Prevalence of smear-positive pulmonary tuberculosis in different ethnic groups in India: evaluation of public health.

OBJECTIVE: To estimate the prevalence of pulmonary tuberculosis (TB) in a high-risk rural area of central India. STUDY DESIGN: Retrospective analysis of primary data. METHODS: In total, 10,963 sputum smears were screened from Hindu tribes (n = 4032), Hindu non-tribal (n = 5445) and Muslim communities (n = 1486) between 2004 and 2009. Smears were recorded as positive or negative for tubercle bacilli following staining with acid-fast bacilli, in accordance with the guidelines of the World Health Organization. Age- and gender-specific prevalence rates and relative risks (RR) were calculated using Statistical Package for the Social Sciences Version 13.0. RESULTS: The prevalence of TB was found to be significantly higher in Hindu tribes compared with Hindu castes and Muslims (P < 0.005). The overall RR of developing smear-positive disease was 1.4-fold higher (95% confidence interval 1.1-1.7; P < 0.005) in males than females in all the study groups. The highest prevalence of TB was observed in subjects aged 15-34 years. CONCLUSIONS: Hindu tribes and males of working age are still at high risk of smear-positive TB.

Coding and non-coding polymorphisms in VDR gene and susceptibility to pulmonary tuberculosis in tribes, castes and Muslims of Central India.

Vitamin D receptor (VDR) plays an important role in activating immune response against various infectious agents. This study was aimed to investigate the association between VDR gene polymorphisms and different clinical forms of pulmonary tuberculosis (TB) in different population groups. Four common polymorphisms (TaqI, ApaI, BsmI and FokI) of VDR gene were studied in clinically diagnosed TB patients and healthy controls from Sahariya tribe (n=377), Bhil tribe (n=95), Chhattisgarh tribe (n=33), general population from North-Central (NC) (n=1021) and South-Eastern (SE) region (n=646) and Muslims (n=217). Genotyping was carried out using PCR-RFLP method and re-confirmed by direct sequencing. The haplotype analysis was performed on Haploview 4.1 and statistical analysis was done using SPSS 13.0 software. We found that bb genotype of BsmI polymorphism conferred significant risk to smear positive and multiple drug resistant (MDR) TB in tribes [OR (CI)=3.7 (1.5-9.2), p=0.002], SE population [OR (CI)=2.1 (1.4-3.3), p=0.0004] and Muslims [OR (CI)=6.7 (1.1-39), p=0.01]. The subjects with FF genotype of FokI polymorphism appeared less likely (p=0.004) to develop MDR TB in NC population, whereas, those with Ff [OR (CI)=2.5 (1.3-5.0), p=0.004] and ff [OR (CI)=3.4 (1.2-9.3), p=0.01] genotypes were at high risk of MDR and smear positive disease, respectively. Similarly, tt genotype of TaqI polymorphism was found associated with high risk of smear positive TB in NC [OR (CI)=3.6 (0.9-14.2), p=0.05] as well as in SE [OR (CI)=4.7 (1.8-12.3), p=0.00003] population. Interestingly, tt genotype appeared strongly associated [OR (CI)=8.9 (2.7-29), p=0.00001] with high bacillary load outcome. In conclusion, genetic polymorphisms in VDR gene, alone or in combination (haplotypes) are associated with different clinical outcomes in pulmonary TB.

Sequence homology and structural analysis of plasmepsin 4 isolated from Indian Plasmodium vivax isolates.

Plasmodium vivax malaria is a globally widespread disease responsible for 50% of human malaria cases in Central and South America, South East Asia and Indian subcontinent. The rising severity of the disease and emerging resistance of the parasite has emphasized the need for the search of novel therapeutic targets to combat P. vivax malaria. Plasmepsin 4 (PM4) a food vacuole aspartic protease is essential in parasite functions and viability such as initiating hemoglobin digestion and processing of proteins and is being looked upon as potential drug target. Although the plasmepsins of Plasmodium falciparum have been extensively studied, the plasmepsins of P. vivax are not well characterized. This is the first report detailing complete PM4 gene analysis from Indian P. vivax isolates. Blast results of sequences of P. vivax plasmepsin 4 (PvPM4) shows 100% homology among isolates of P. vivax collected from different geographical regions of India. All of the seven Indian isolates did not contain intron within the coding region. Interestingly, PvPM4 sequence analysis showed a very high degree of homology with all other sequences of Plasmodium species available in the genebank. Our results strongly suggest that PvPM4 are highly conserved except a small number of amino acid substitutions that did not modify key motifs at active site formation for the function or the structure of the enzymes. Furthermore, our study shows that PvPM4 occupies unique phylogenetic status within Plasmodium group and sufficiently differ from the most closely related human aspartic protease, cathepsin D. The analysis of 3D model of PM4 showed a typical aspartic protease structure with bi-lobed, compact and distinct peptide binding cleft in both P. vivax and P. falciparum. In order to validate appropriate use of PM4 as potential anti-malarial drug target, studies on genetic and structural variations among P. vivax plasmepsins (PvPMs) from different geographical regions are of utmost importance for drugs and vaccine designs for anti-malarial strategies.

In vitro induction of the ubiquitous 60 and 70KD heat shock proteins by pesticides monocrotophos and endosulphan in Musca domestica: potential biomarkers of toxicity.

This study has investigated the effect of two highly toxic pesticides, monocrotophos (organophosphate) and endosulphan (organochlorine), on the inducibility of two major heat shock proteins, the HSP60 and HSP70, essential for cell survival, in the house fly Musca domestica. The LC50 values of the two pesticides for larva and adult (monocrotophos: 0.05 ppm for larva and 0.025 ppm for adult; endosulphan: 15 ppm for larva and 2 ppm for adult) revealed monocrotophos to be potentially more toxic than endosulphan. The relative susceptibility (lethality) of adult to either of these pesticides appeared much higher than that of larva. The expression patterns of HSP60 and HSP70 were analysed in various larval and adult tissues, exposed to varying sub-acute doses of monocrotophos (0.00010 ppm - 0.00075 ppm for larva and 0.00010 ppm - 0.00050 ppm for adult) and endosulphan (0.5ppm - 2.0ppm for larva and 0.01ppm-0.10 ppm for adult). The immunoblots revealed significant correlation between the pattern of HSP's expression and the pesticides-induced tissue injury/ mortality, visualized by trypan blue dye exclusion test. Both the pesticides caused significant induction of these HSPs in a tissue and dose­dependent manner, suggesting their importance as molecular indicators (biomarker) in the assessment of cellular toxicity caused by endosulphan and monocrotophos.

Genomic structure and sequence analysis of Lucilia cuprina HSP90 gene.

The HSP90 family is one of the highly conserved chaperone families, varying between eubacteria to higher vertebrates. The HSP90 protein has been assigned different functions including thermal protection, but having major role in development. The present study is a detailed analysis of the structural characteristics of hsp90 gene (lchsp90) of sheep blowfly, Lucilia cuprina. The gene isolated by PCR revealed absence of intron. The nucleic acid and amino acid comparison revealed significant level of sequence similarity among species of various taxa. Significantly, the analysis of the amino acid sequence revealed that the HSP90 of L. cuprina belongs to hsp90ß class.

Evaluation of mutagenic and antimutagenic activities of neem (Azadirachta indica) seed oil in the in vitro Ames Salmonella/microsome assay and in vivo mouse bone marrow micronucleus test.

AIM OF THE STUDY: The possible mutagenic and antimutagenic activity of neem oil (NO) and its DMSO extract (NDE) were, examined in the Ames Salmonella/microsome mutagenicity test and the mouse bone marrow micronucleus assay. MATERIALS AND METHODS: Eight different strains of Salmonella typhimurium were, used to study the genotoxicity of neem oil both in the presence and absence of Aroclor-1254 induced rat liver homogenate (S9). Two-dose treatment protocol was, employed to study the cytogenetic activity in micronucleus assay. Similarly, the antimutagenic activity of neem oil and NDE was studied against mitomycin (MMC) and 7,12-dimethylbenz[a]anthracene (DMBA) in the above two test systems. RESULTS: Neem oil was non-mutagenic in all the eight tester strains of Salmonella typhimurium both in the presence and absence of S9 mix. In the present study, there was no significant increase in the frequency of micronucleated polychromatic erythrocytes (MNPCEs) in neem oil treated groups over the negative control (DMSO) group of animals, indicating the non-clastogenic activity of neem oil in the micronucleus test. Neem oil showed good antimutagenic activity against DMBA induced mutagenicity compared to its DMSO extract. However, neem oil showed comparatively less antimutagenicity against MMC in the Ames assay. In vivo anticlastogenic assays shows that neem oil exhibited better activity against DMBA induced clastogenicity. CONCLUSION: These results indicate non-mutagenic activity of neem oil and significant antimutagenic activity of neem oil suggesting its pharmacological importance for the prevention of cancer.

DAZL 260A > G and MTHFR 677C > T variants in sperm DNA of infertile Indian men.

DAZL (deleted in azoospermia-like) 260A > G and MTHFR (methylene tetrahydrofolate reductase) 677C > T are two important autosomal variants associated with impaired spermatogenesis. In this study, we investigated DAZL 260A > G and MTHFR 677C > T variants in sperm DNA and their frequency in oligozoospermic infertile men of Indian origin. The study on sperm DNA was performed, since it is more prone to oxidative stress-induced damage and mutation. One hundred oligozoopsermic infertile men having normal chromosomal complement with intact Y chromosome and 100 age- and ethnically-matched fertile controls were investigated for these variants in their sperm genome. Spermatozoa were separated by gradient centrifugation and DNA was isolated and analyzed for the single nucleotide polymorphisms (SNPs) by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). The results showed no significant differences in the frequency of DAZL AG (P = 0.58) and MTHFR CT (P = 0.44) between oligozoospermic infertile men and controls. However, 8% (8/100) oligozoospermic infertile men harbored both the variants and showed significantly (P < 0.0001) lower sperm count (3.28 +/- 1.1 vs 12.50 +/- 4.09) compared to infertile men with either of the single variant. None of the fertile controls showed the presence of the both variants. In conclusion, the combined effect of both DAZL 260A > G and MTHFR 677C > T variants may have role in compromised sperm count. However, further studies are required to find the pathological role of these combined variants in male infertility.

Utility of serum LDH isoforms in the assessment of mycobacterium tuberculosis induced pathology in TB patients of Sahariya tribe.

The present study was carried out in the Sahariya tribe of Central India, which reportedly have high prevalence of pulmonary tuberculosis. Total serum LDH and its tissue specific isoforms were estimated in TB patients and matched healthy controls to test the utility of LDH as diagnostic marker for tuberculosis. About 210 sputum positive cases and 328 age and sex matched sputum negative controls were recruited. The spectrophotometeric and densitometric analysis of each LDH isoform was carried out in both cases and controls. The mean values of serum LDH were estimated and compared for each class by t-test. The statistical comparisons were made between sputum negative controls and sputum positive cases by Mann-Whitney's U test. The spectrophotometric estimation of serum LDH revealed significant (P=0.0016) increase in its level in cases (290 IU/L) as compared to controls (248 IU/L). The densitometric analysis of individual LDH isoforms in cases and controls demonstrated significant elevation in LDH1 (P>0.05), LDH2 (P>0.05) and LDH3 (P<0.005) in sputum positive cases in comparison to sputum negative controls. Our study revealed a positive correlation between serum LDH level and the presence of mycobacteria and their load, suggesting utility of LDH as an important diagnostic marker of tuberculosis induced stress, at least in tribal areas lacking access to modern clinical tests.

In vitro induction of 60-kDa and 70-kDa heat shock proteins by endosulphan and monocrotophos in sheep blowfly Lucilia cuprina.

The effect of two most commonly used and highly toxic organic pesticides, endosulphan (organochlorine) and monocrotophos (organophosphate), was studied in a blowfly, Lucilia cuprina, to test whether these pesticides induce the stress response and, if so, whether the intensity of the response, in terms of induction of heat shock proteins (HSPs), HSP60 and HSP70 in particular, is pesticide concentration dependent. The in vitro exposure of larval and adult tissues to varying concentrations of these pesticides (endosulphan: 1.0-4.0 ppm for larva and 0.05-0.50 ppm for adult; monocrotophos: 0.0005-0.0050 ppm for larva and 0.0001-0.0010 ppm for adult) revealed that both compounds were able to induce the expression of HSP60 and HSP70 proteins. Western blot analysis of these HSPs indicated that the induction of expression was tissue-specific. The trypan blue staining of pesticide-exposed tissues demonstrated monocrotophos to exert more severe effect than endosulphan, as the former compound induced both HSP60 and HSP70 significantly at a much lower concentration than that of the later. The pattern of expression of these HSPs, in general, appeared in direct correlation with the pesticide concentration. Gut tissues were found relatively more sensitive to pesticide toxicity than other tissues, as revealed by trypan blue staining, and hence, they might serve as primary targets for early detection of pesticide toxicity. The results indicated that either of these HSPs or both could serve as a potential biomarker toward assessment and monitoring of toxicity induced by these pesticides.