Assessment of occupational exposure to diesel particulate matter through evaluation of 1-nitropyrene and 1-aminopyrene in surface coal miners, India.

DPM (diesel particulate matter) is ubiquitously present in the mining environment and is known for mutagenicity and carcinogenicity to humans. However, its health effects in surface coal mines are not well studied, particularly in India. In this study, DPM exposure and corresponding exposure biomarkers were investigated in four different surface coal mines in Central India. To document and evaluate the DPM exposure in surface coal miners, we characterized 1-NP (1-nitropyrene) in the mining environment as surrogate for DPM using Sioutas Cascade Impactor. Exposure biomarkers were analyzed by collecting post work shift (8-h work shift) urine samples and determining the concentrations of 1-aminopyrene (1-AP) as a metabolite of 1-NP and 8-hydroxydeoxyguanosine (8OHdG) as DNA damage marker. We observed high concentration of 1-NP (7.13-52.46 ng/m3) in all the mines compared with the earlier reported values. The average creatinine corrected 1-AP and 8OHdG levels ranged 0.07-0.43 [Formula: see text]g/g and 32.47-64.16 [Formula: see text]g/g, respectively, in different mines. We found 1-AP in majority of the mine workers' urine (55.53%) and its level was higher than that reported for general environmental exposure in earlier studies. Thus, the study finding indicates occupational exposure to DPM in all the four mines. However, the association between 1-NP level and exposure biomarkers (1-AP and 8OHdG) was inconsistent, which may be due to individual physiological variations. The data on exposure levels in this study will help to understand the epidemiological risk assessment of DPM in surface coal miners. Further biomonitoring and cohort study are needed to exactly quantify the occupational health impacts caused by DPM among coal miners.

Peripheral assessment of the genes AQP4, PBP and TH in patients with Parkinson's disease.

Parkinson's disease (PD) typically appears in late middle-aged and in elderly persons progressing over a period of several years. The characteristic pathological features of PD patients include defective motor function and cognitive function affecting the quality of life of PD patients. Oxidative stress is considered to a play a central role along with various other factors in the pathogenesis of PD and the incidence and prevalence of the disease is incessantly increasing worldwide. The objective of the current study was to assess mRNA expressional changes of AQP4, TH and PBP in blood samples of control and patients with PD. The study included 30 healthy controls and 90 PD patients subjected to treatment through the entire period of the study. RNA isolation was carried out using blood samples of the subjects recruited in the study and used for RT-PCR analysis of AQP4, TH as well as PBP. The mRNA expressions of AQP4 and TH were found to be reduced whereas that of PBP was found to be elevated when compared with those of healthy control samples. The statistically analysed data were presented which could be helpful for appreciation of PD pathology reflecting in the blood samples of PD population.

Studies on the neuroprotective role of Piper longum in C6 glioma induced rats.

Many naturally occurring substances of plant origin ingested in human diet, exhibit anticarcinogenic and antimutagenic effects. One of the active phytochemical which shows the active anticarcinogenic role is Piper longum Linn. (Pl). Pl is widely used in ayurvedic industry due to its property in healing some of the bodily ailments. Despite being known for the antioxidant, antimicrobial and anticarcinogenic effects, its relation to brain and its tumour development is still scarce. Hence, the experimental glioma model was developed in rats using C6 glioma cells and the effect of Pl was evaluated in the brain tissue of experimental group of rats. From the study, the glioma induced animals showed an increased level of lipid peroxides (LPO), tissue marker enzymes lactate dehydrogenase (LDH), creatine kinase (CK), 5'nucleotidase (5'ND) and acetylcholine esterase (AChE). But Pl treatment (20 mg/kg body weight) significantly attenuated these alterations thereby showing potent anticancer effect in glioma induced rats. In addition, the anticarcinogenic effect of Pl was confirmed by microscopic analysis and the restoration of increased lipids and protein bound carbohydrates (PBCs) in the brain tissue of glioma induced rats. Hence our results implicate a major role for Pl in preventing the cancer development in the experimental glioma model.

Neurotrophin Expression in Lymphocytes: a Powerful Indicator of Degeneration in Parkinson's Disease, Amyotrophic Lateral Sclerosis and Ataxia.

Deregulated neurotrophin is an etiological factor in the pathology of neurodegenerative diseases (ND) that are clinically different entities but characterised by similar limb dysfunction. Earlier validation of peripheral biomarkers can provide significant translational benefit to ND patients. We analysed brain-derived neurotrophic factor (BDNF)-tropomyosin possessing tyrosine-related kinase (Trk B) and its key downstream proteins which are implicated in ND such as Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS) and ataxia. Blood from ND patients with PD, ALS and Ataxia with movement dysfunctions were obtained to analyse mRNA and protein expressions of the above mentioned factors in lymphocytes. The mRNA and protein expression of BDNF-Trk B and its key downstream molecules showed a significant variation when compared to control and among NDs. The study intends to show that on identifying the variation of these key molecules in the blood samples of patients with ND can serve as early diagnostic candidates. Thus by intervening, the neurotrophins and their pathways can help in early diagnosis and optimising levels of diagnostic certainty.

The effect of N-acetylcysteine in combination with vitamin C on the activity of ornithine decarboxylase of lung carcinoma cells--In vitro.

Ornithine decarboxylase (ODC) is a marker of lung cancer and is a key enzyme in the synthesis of polyamines, which are necessary for the promotion of the growth of malignant cells. This study assesses the dose-dependent effect of N-acetylcysteine (NAC), a chemopreventive agent, in combination with vitamin C (VC) on the activity of ODC in lung carcinoma cell line, NCI-H82. The cells were subjected to supplementation of NAC and VC both individually and in combination at different dosages for 24 h as well as 48 h. The cells were incubated with radiolabeled L-ornithine (14C) after the supplementation of NAC and VC individually as well as in combination. A microprocedure was carried out to estimate the activity of ODC in cells after 24 and 48 h of incubation. The activity which was found to be elevated in control cells was decreased significantly on drug supplementation in dose-dependent fashion. The content of nucleic acids also exhibited similar result and [3H]-thymidine incorporation was also affected by the supplementation.

Neurotransmissional, structural, and conduction velocity changes in cerebral ganglions of Lumbricus terrestris on exposure to acrylamide.

Acrylamide (ACR), an environmental toxin though being investigated for decades, remains an enigma with respect to its mechanism/site of actions. We aim to explicate the changes in cerebral ganglions and giant fibers along with the behavior of worms on ACR intoxication (3.5-17.5 mg/mL of medium/7 days). Neurotransmitter analysis revealed increased levels of excitatory glutamate and inhibitory gamma amino butyrate with reduced levels of dopamine, serotonin, melatonin, and epinephrine (p < 0.001). Scanning electron microscopy showed architectural changes in cerebral ganglions at 3.5 mg/mL/ACR. The learning behavior as evidenced by Pavlovian and maze tests was also altered well at 3.5 mg/mL of ACR. Electrophysiological assessment showed a reduction in conduction velocity of the medial and lateral giant nerve fibers. We speculate that the observed dose/time-dependent changes in neurotransmission, neurosecretion, and conduction velocity on ACR intoxication at 17.5 mg/ml, possibly, could be due to its effect on nerve fibers governing motor functions. The bioaccumulation factor in the range of 0.38-0.99 mg/g of ACR causes a detrimental impact on giant fibers affecting behavior of worm. The observations made using the simple invertebrate model implicate that the cerebral ganglionic variations in the worms may be useful to appreciate the pathology of the neurological diseases which involve motor neuron dysfunction, esp where the availability of brain samples from the victims are scarce.

Quality of life measures in glioma patients with different grades: A preliminary study.

BACKGROUND: Plethora of information exists in the literature on pathology of the glioma while prevailing research data on quality-of-life (QOL) of glioma patients marks dearth thus demanding more studies. AIMS: In this study, we examined the QOL of different grades of glioma patients among the Chennai population in India. MATERIALS AND METHODS: A total of 162 patients with different grades of glioma enrolled from August 2007 to February 2011, at their first contact to Department of Neurology and Neurosurgery, Government General Hospital, Chennai, India were included and their QOL was assessed by European Organization for Research and Treatment of Cancer Core QOL questionnaire (EORTC QLQc-30), EORTC brain cancer module (QLQ BN-20). RESULTS AND CONCLUSION: Both low and high grade glioma (LGG and HGG) patients had poor mean scores in social functioning (87.0), physical functioning (82.0) and emotional functioning (75.2) and role functioning (58.9). The mean scores on cognitive functioning (61.9) and global QOL (60.3) were better. Age, Karnofsky performance status, World Health Organization grades showed significant associations with all functional scales. The percentage values were higher for symptoms of fatigue (76.9%), pain (71.5%), financial difficulties (77.6%) and appetite loss (38.46%) in both LGG and HGG. Similarly, with respect to QLQ-BN20 domains, HGG patients showed more symptoms than low grade with a significant correlation in communication deficit problems (P = 0.02), headache (P = 0.04), seizures (P < 0.01), hair loss (P < 0.05) than the other symptoms. This initial assessment suggests that an increasing burden of symptoms exists, with poor QOL and survival, which has become a major concern in different grades of glioma patients.

Status of lipid peroxidation and antioxidant enzymes in malignant (bronchogenic carcinoma) and non-malignant pleural effusions.

Eighty patients from Chennai Medical College (patients with bronchogenic carcinoma) and from Tambaram Tuberculosis Hospital (patients with non-malignant pulmonary diseases mainly tuberculosis) in whom the etiologic diagnosis of their pleural effusions are confirmed were included in the study. Lipid peroxidation (LPO) and activities of antioxidant enzymes were estimated in pleural exudates of the two groups. Lipid peroxidation was found to be increased and the status of antioxidants were found to be decreased in lung malignant pleural exudates when compared to those of non-malignant effusions. The possible reasons for the observed results discussed.

A study on the biochemical and cytogenetic status in the blood of glioma patients.

Malignant gliomas, the most common subtype of primary brain tumors, are aggressive, highly invasive and neurologically destructive tumors, considered being the deadliest of human cancers. As an attempt to understand the biology of glial tumor, a study on macromolecules like proteins, matrix metalloproteinases, lipids antioxidants and Deoxyribonucleic acid, in the blood of glioma patients was made. Biochemical assessment of significant pathophysiological enzymes, antioxidants and marker enzymes was performed. MMP expression was determined using gelatin zymography. Karyotyping analysis was done to determine chromosomal aberrations. A marked rise was observed in the proteins and lipids of glioma patients as compared to the normal cases. The antioxidant status of the patients was found to be lowered. Karyotypic analysis of the peripheral blood chromosomes presented various chromosomal aberrations in glioma patients. The biochemical parameters were significantly increased in the patient population (p<0.01, p<0.001) when compared to those of normal. Zymographic analysis showed the presence of MMP-2 and MMP-9 in the patient sample. Karyotypic investigation showed alterations in the chromosomal pattern of the glioma patients. The study provides baseline information on the biochemical alterations in the blood of glioma patients which can be further exploited for detailed investigations.

Molecular interaction of Survivin and Piperine by computational docking analyses for neuroblastoma targeting.

BACKGROUND: Neuroblastoma (NB) is a childhood cancer causing significant mortality in at least 1% children worldwide. NB is an embryonically derived tumor. The causative agents include genetic predisposition and dysregulated signaling cascades. Survivin is an important anti-apoptotic protein that is significantly up-regulated in NB. In this study, a naturally occurring ligand - Piperine was assessed for its interaction with Survivin protein. PURPOSE: The study was undertaken in order to identify the experimental feasibility of Survivin inhibitor ligand Piperine as targeting treatment of NB. METHODS: Protein sequences were retrieved and saved in PDB format. Similarly, the ligand data was processed using MGL (Molecular Graphics Laboratory) and chimera tools and saved in PDB format. Both protein and the ligand data were then uploaded to the docking server and docking parameters were set. RESULTS: In-silico docking study of a protein ligand interaction resulted in -3.36 Kcal/mol free energy value for the ligand, with an involvement of 1 hydrogen bond, 7 hydrophobic interactions and 13 ionic interactions. The results were correlated with the existing free energy value of > -3 Kcal/mol which is established for a good inhibitor. CONCLUSION: The molecular docking study for mice Survivin and Piperine shows good inhibitory interaction effect and can, therefore, be considered as a molecule against Survivin enhanced tumor condition including NB.

Oxidative pathology and AQP4 mRNA expression in patients of Parkinson's disease in Tamil Nadu.

BACKGROUND: Parkinson's disease (PD) typically appears in late middle aged and elderly persons and progresses over a period of several years. It is characterised by defective motor and cognitive function. Oxidative stress is believed to play a central role in the pathogenesis of PD. PURPOSE: The objective of the study was to assess the oxidative burden and mRNA expression of AQP4 related to oxidative pathology of PD related symptoms of Hoehn and Yahr stages. METHODS: The study included 30 healthy controls and 90 PD patients who were undergoing treatment. The blood samples were collected and analyzed for biochemical assays and whole blood DNA was used for mRNA expression of AQP4 using RT-PCR. RESULTS: The level of SOD, CAT and Gpx were found to be decreased while there was increase in LPO when compared to the healthy controls. The levels of SOD, CAT in stage III were significantly decreased when compared with stage I. The mRNA expression of AQP4 was found to be reduced when compared with that of healthy control samples. There was no variation in observed oxidative burden and the AQP4 mRNA expression among the different stages of disease. CONCLUSION: Based on the results obtained this study may be helpful in validating novel approach to treatment of PD by advancing antioxidant strategies.

Frequency variations in the methylated pattern of p73/p21 genes and chromosomal aberrations correlating with different grades of glioma among south Indian population.

Gliomas are the most common primary brain tumors in India. The main epigenetic modification in glioma is aberrant DNA methylation that is now renowned to be a common hallmark of brain tumors. This study was designed to determine the frequency of aberrant CpG island methylation in the promoter regions of p21 and p73 in different grades of glioma and to explore their respective chromosomal aberrations. Total of 160 patients with histologically confirmed grades of glioma (I, II, III, and IV) were included in the study. DNA samples from blood and brain tissues, including benign lesions were subjected to sodium bisulfite conversion and hypermethylation detection using methylation-specific PCR followed by RT-PCR. Western blotting was also carried out for p21 and its related protein, p53. A total of 124 of 160 glioma samples (77.5%) displayed CpG island hypermethylation of both p73, p21 genes associated with the loss of mRNA expression (P < 0.001) and the loss of protein expressions (p53 independent p21 expression). p73 gene showed increased methylation frequency in all grades, 40 of 60 (66%) glioblastomas and 16 of 30 (53.3%) anaplastic astrocytoma, 10 of 20(50%) oligodendrogliomas, 8 of 20 (40%) ependymoma, and low-grade glioma 6 of 20 (30%). The percentage of methylation significantly well correlated with the overall survival and also with chromosomal loss. Thus, the studied glioma patients among south Indians showed a high frequency of aberrant methylation with varied chromosomal signatures in different grades, playing a role in aggressiveness and characterization of a particular grade, the appreciation of which might help for designing a specific therapy.

Investigations on light induced stress model and on the role of Phyllanthus amarus in attenuation of stress related depression with focus on 5HT2Am- RNA expression.

BACKGROUND: There is an increasing evidence that psychological stress and depression trigger changes in various biochemical parameters in animals and human subjects. Chronic stress in rats, and psychosocial stress in humans, is implicated in the pathophysiology of mood and anxiety disorders. PURPOSE: The current study was been designed to investigate the behavioral, anatomical status of rat brain and expression of serotonin receptor (5HT2A) mRNA related to pathophysiology of stress after high light (HL) illumination and to evaluate the effect of Phyllanthus amarus (PA) over such stress induced changes in mid brain and prefrontal cortex region of rat brain. METHODS: Bright light illumination was used to induce stress in wistar rats. Established methods were used to analyse biochemical and histopathological tests. RESULTS: PA administration to HL stressed animals for 7 days prevented stress-induced oxidative damage, as evidenced by significant enhancement of key endogenous antioxidant defense components. HL stress also caused reduced activities of membrane bound enzymes in synaptic membrane along with reduced levels of lipid profile and glycoprotein in the midbrain region. Stress induced changes in the locomotor activity, time spent for exploring the center of the arena, frequency of rearing and grooming, and frequency of facing the corner of the arena, were altered on PA administration. CONCLUSION: The findings suggest that the therapeutic efficacy of PA may be mediated, atleast partially, via reversal of oxidative damage. Further the study demonstrates the promising intervention by PA on the mRNA expression of 5HT 2A in the brain. These preliminary results pave the way for further validation of PA against stress.

Curcumin combats against cigarette smoke and ethanol-induced lipid alterations in rat lung and liver.

BACKGROUND: Human population, in spite of the medical and scientific achievements, still fall as a prey to the evils of habitual smoking and alcohol, thus necessitating safer counteracting measures. OBJECTIVE: To evaluate the effect of cotreatment of curcumin (Curcuma longa) in rats subjected to acute exposure to cigarette smoke (CS) and ethanol (EtOH). METHODOLOGY: Of the four groups of experimental rats, a set of rats was subjected to whole body exposure to cigarette smoke along with ethanol administration serving as a model of CS+EtOH injury. Curcumin treatment was given to two sets of rats: (i) one set receiving simultaneous CS+EtOH and (ii) one set of normal rats without any administration. The other group of rats served as control. Blood, liver and lung of rats were selected for assessment of CS+EtOH injury as well as curcumin treatment. RESULT: Altered lipid, lipoprotein profile and bile acid excretion were observed in CS+EtOH rats along with premalignant pathological state in tissues. In treated rats, the levels were maintained at near-normal levels along with near-normal histology. CONCLUSION: This biochemical picture on cotreatment with curcumin suggests that curcumin could counteract the injurious effects of combined CS and EtOH and thus might help to reduce the risk of hyperlipidemic disorders which develop due to smoking and drinking.

Histone H1 inhibits the proliferation of MCF 7 and MDA MB 231 human breast cancer cells.

Purified histone H1 exerts extracellular functions suggesting novel histone functions. The cytotoxic effects of histone H1 have lead to its choice as a pharmacological tool in breast cancer. Hence the present study was aimed at investigating the effect of exogenous histone H1 on the proliferation of estrogen receptor positive (MCF 7) and estrogen receptor negative (MDA MB 231) human breast cancer cells. Cells were incubated with various concentrations of histone H1 and antiproliferative activity was assessed by MTT assay. Proliferation of breast cancer cells was assessed from the activity of ornithine decarboxylase (ODC) using [(14)C] labeled ornithine. Histone H1-mediated cellular effects, such as anchorage dependent growth and apoptosis, were assessed by colony formation assay, fluorescence microscopy after acridine orange/propidium iodide staining and DNA fragmentation analysis. Histone H1 was significantly cytotoxic as it inhibited colony formation, ODC activity and induced apoptosis in both estrogen receptor positive and estrogen receptor negative cells. These results suggest that histone H1-induced antiproliferative effects on human breast cancer cells could possibly involve inhibition of ODC.

The antiulcer activity of Garcinia cambogia extract against indomethacin-induced gastric ulcer in rats.

Garcinia cambogia extract is a herbal preparation that has been suggested as useful in the treatment of gastrointestinal disorders. In the present study this drug was tested for its antiulcerogenic effect. Oral pretreatment with Garcinia cambogia fruit extract (1 g/kg body wt/day) for 5, 10 or 15 days protected the gastric mucosa against the damage induced by indomethacin (20 mg/kg body wt). The volume and acidity of the gastric juice decreased in the pretreated rats. The glycoprotein levels of the gastric contents which were decreased in the untreated rats, maintained near normal levels in the pretreated rats. Protein which was elevated in the gastric juice of untreated rats, showed near normal levels in the pretreated rats. Garcinia cambogia was able to decrease the acidity and to increase the mucosal defence in the gastric areas, thereby justifying its use as an antiulcerogenic agent.