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Cigarette smoke is responsible for multiple disorders and causes almost 10 million annual deaths globally but underlying mechanisms are still underexplored. Continuous exposure of Cigarette smoke condensate (CSC) leads to cytosolic phospholipase A2 (cPLA2) mediated high free radicals where cPLA2s seems to play crucial role in generated various patho-physiological conditions such as chronic inflammation, oxidative stress and cancer. In this view, we assessed the therapeutic potential of arachidonyl trifluromethyl ketone (ATK), a cPLA2 inhibitor, via pharmacological inhibition of most expressible CSC-induced cPLA2 group IVA in type-I and type-II alveolar epithelial cells. The In Vitro inhibitory effect of ATK on CSC-induced PLA2 activity and its cellular role were assessed in terms of cell viability, fluorescein diacetate (FDA) dye uptake assay for membrane integrity, reactive oxygen species (ROS)/reactive nitrogen species (RNS) levels and pro apoptotic as well as anti apoptosis markers via flow cytometry, along with extracellular signal-regulated kinases (ERK) levels using enzyme-linked immunosorbent assay (ELISA). The experimental findings demonstrated that ATK acts as potent inhibitor of cPLA2 activity and shown its effectiveness as therapeutic agent by significantly mimicking CSC-induced levels of free radicals, primary apoptosis, ratio of pro-apoptotic/apoptotic proteins and levels of ERK whereas protected cells from loss of cell viability and membrane integrity. Thus, this study is an important step towards the opening up of avenues for the applicability of the cPLA2 isoform specific inhibitors such as ATK for pre-clinical and clinical studies and could be beneficial during smoking-induced lung pathological conditions.

Background: Colorectal cancer (CRC) is the third most common cancer prevalent in men, and the second in women worldwide. The exact cause and pathogenesis of this disease remains unknown. In every cell multiple phospholipase A2 (PLA2) exists, but their contribution to the cell function has yet not been determined. The increased activity of these isoforms may be important in the generation of inflammatory mediators which finally are involved in the initiation of carcinogenesis. This study has explored the pathways involved in the differential response of HCT-15 and HT-29 colon cell lines during benzo(α)pyrene [B(α)P]-induced molecular changes.Methods: HT-29 and HCT-15 cells were used for the present study. Benzo(α)pyrene [B(α)P]-induced molecular changes were evaluated through cell viability using MTT assay, reactive oxygen species (ROS) measurement using 2,7 dichloro-dihydro-fluorescin diacetate (DCFH-DA), The gene expressions of PLA2 isoforms were estimated by RT- PCR. Further, knockdown of PLA2 isoform in transfected cells was done with siRNA.Results: The cell viability decreased significantly after exposure with B(α)P in both cell lines. Reactive oxygen species (ROS) production after exposure with B(α)P was significantly induced in both types of colon cell lines. The gene expressions of PLA2 isoforms were also estimated. Three PLA2 isoforms such as IB and, IID, IVA were induced after exposure with B(α)P respectively in HT -29 and HCT-15 cell lines. Further, a significant knockdown of PLA2 isoform in transfected cells with siRNA, and B(α)P exposure was observed in HCT-15 and HT-29 cells. There was a significant induction of ROS in IVA transfected HCT-15 cells exposure with B(α)P.Conclusion: It is concluded that each cell line behaved differently, specific PLA2 siRNA are involved in controlling the cascades related ROS, depending upon the origin of the cells. Secretary phospholipase A2 (sPLA2) inhibitors are likely to have a therapeutic potential in colon pathologies.

Cigarette smoke (CS) is a complex combination of over 5000 compounds divided into two phases (tar and gas) whose exposure causes various lung pathological conditions especially COPD and cancer. In a single puff, tar phase basically contains extremely high concentrations of stable and long-lived radicals (~1017 radicals/gm), whereas gas phase contains ~1015 organic radicals. In contrast to stable life radicals (e.g., hydrogen peroxide, hypochlorous acid) present in tar phase, the radicals in gaseous phase are highly reactive nitrogen and oxygen-centered radicals (e.g., nitric oxide, reactive olefins, dienes) known to have lifetime of less than 1 s. Therefore, cigarette smoke constituents evoke the endogenous oxidants production and agitate normal functioning of tissues, especially type I and II pulmonary epithelium. As a result of smoke exposure, increased level of oxidants (exogenous or endogenous) leads to stress state termed as “oxidative stress,” which arises due to commotion in prooxidant and antioxidant balance in favor of the prooxidant. The oxidative stress caused by the constituents of cigarette smoke in normal as well as in the mutated cells leads to activation of various upstream signaling events such as activation of lipid-specific enzymes especially phospholipases whose role is crucial in the regulation and remodeling of membrane lipids when subjected to hazardous and noxious environmental insults such as cigarette smoke-initiated tissue damage.

Tobacco consumption globally is estimated to be around 20% and in India accounted for 12% of the world’s smokers according to the World Health Organization (WHO) statistics. It is also estimated that around six million deaths occur every year and rises to eight million by the year 2030. The socioeconomic health burden is of great concern due to the rise in cigarette smoking and related deaths. Cigarette smoking is known to cause many disorders such as chronic obstructive pulmonary diseases (COPD), oral cancers, and pulmonary tumors. More than 6000 different chemicals contain approximately 100 known pulmonary carcinogens present in the cigarette smoke (CS). Inhaled polycyclic aromatic hydrocarbons (PAHs), constituents of CS, are deposited in the airway epithelium and the majority in the alveolar epithelium which detoxifies the cigarette constituents by way of drug-metabolizing enzymes such as cytochrome p450 (CYP450) enzymes. Reactive oxygen species (ROS) present in the CS and the CS-mediated generation of mitochondrial and cellular ROS have an impact on the cellular bio-constituents such as membrane lipids, proteins, and DNA. Studies suggest that oxidative stress causes lung inflammation by remodeling of membrane lipids and pulmonary surfactants via activation of phospholipase A2 (PLA2), cyclooxygenases (COX), and lipoxygenases which directly or indirectly activates proinflammatory cytokines. Considering the complexity of lung tissue and functions, understanding the cellular and tissue mechanisms of damage and strategies overcoming the deleterious effects mediated by the CS would be prime importance. This understanding helps target specific molecules that will help in developing therapeutic strategies to circumvent the CS-mediated disease burden, mortality, morbidity, and economic burden.

Cigarette smoke is responsible for multiple disorders and causes almost 10 million annual deaths globally but underlying mechanisms are still underexplored. Continuous exposure of Cigarette smoke condensate (CSC) leads to cytosolic phospholipase A

Background Inflammatory bowel disease (IBD) is an idiopathic, chronic inflammatory condition, which affects the gastrointestinal tract and has no curative treatment. The present study aimed to investigate the effect of different doses of pioglitazone alone and in combination with sulfasalazine in TNBS (trinitrobenzenesulfonic acid)-induced inflammatory bowel disease (IBD) in rats. Methods A total of 36 animals were included in the study. Animals were divided into five groups (n = 6): group I – vehicle (ethanol), group II – TNBS + ethanol, group IIIA–TNBS + pioglitazone (15 mg/kg), group IIIB – TNBS + pioglitazone (30 mg/kg), group IV – TNBS + sulfasalazine (360 mg/kg), group V – TNBS + sulfasalazine (360 mg/kg) + pioglitazone (least effective dose found in group III). Group III was divided into two subgroups, namely ULA and IIIB, on the basis of different doses of pioglitazone used. After completion of two weeks of treatment, rats were sacrificed under ether anesthesia by cervical dislocation for assessment of intestinal inflammation, histological analysis, myeloperoxidase assay, malondialdehyde assay and TNF-α estimation. Results All the drug-treated groups showed both gross morphological and microscopic score either 1 or 2. None of them showed score of > 2 on both gross and microscopic morphological examination. Both MDA levels and MPO activity were significantly re-duced in the drug-treated groups, with maximum reduction seen in the combination group. TNF-α was reduced in pioglitazone group. It was highly reduced in sulfasalazine group (group V) as compared to TNBS group thereby indicating that pioglitazone is protective in TNBS-induced inflammatory bowel disease. Conclusion The present study showed reduction in lipid peroxidation, malondialdehyde levels and TNF-α levels in pioglitazone-treated group and hence, there was significant improvement in gross and microscopic features, too. However, combination of pioglitazone and sulfasalazine has shown greater efficacy.

Background Smoking is one of the leading causes of millions of deaths worldwide. During cigarette smoking, most affected and highly exposed cells are the alveolar epithelium and generated oxidative stress in these cells leads to death and damage. Several studies suggested that oxidative stress causes membrane remodeling via Phospholipase A2s but in the case of cigarette smokers, mechanistically study is not yet fully defined. In view of present perspective, we evaluated the involvement of cytosolic phospholipase A2 (cPLA2) IVA as therapeutic target in cigarette smoke induced pathologies in transformed type I and type II alveolar epithelial cells. Methods Transformed type I (WI26) and type II (A549) alveolar epithelial cells were used for the present study. Cigarette smoke condensate (CSC) was prepared from most commonly used cigarette (Gold Flake with filter) by the Indian population. CSC-induced molecular changes were evaluated through cell viability using MTT assay, reactive oxygen species (ROS) measurement using 2,7 dichlorodihydrofluorescin diacetate (DCFH-DA), cell membrane integrity using fluorescein diacetate (FDA) and ethidium bromide (EtBr) staining, super oxide dismutase (SOD) levels, cPLA2 activity and molecular involvement of specific cPLA2s at selected 24 h time period. Results CSC-induced response on both type of epithelial cells shown significantly reduction in cell viability, declined membrane integrity, with differential escalation of ROS levels in the range of 1.5–15 folds and pointedly increased cPLA2 activity (p

Folic acid is an essential nutrient that is required for one-carbon biosynthetic processes and for methylation of biomolecules. Deficiency of this micronutrient leads to disturbances in normal physiology of cell. Chronic alcoholism is well known to be associated with folate deficiency, which is due in part to folate malabsorption. The present study deals with the regulatory mechanisms of folate uptake in liver during chronic alcoholism. Male Wistar rats were fed 1 g/kg body weight/day ethanol (20 % solution) orally for 3 months, and the molecular mechanisms of folate uptake were studied in liver. The characterization of the folate transport system in liver basolateral membrane (BLM) suggested it to be a carrier mediated and acidic pH dependent, with the major involvement of proton coupled folate transporter and folate binding protein in the uptake. The folate transporters were found to be associated with lipid raft microdomain of liver BLM. Moreover, ethanol ingestion decreased the folate transport by altering the Vmax of folate transport process and downregulated the expression of folate transporters in lipid rafts. The decreased transporter levels were associated with reduced protein and mRNA levels of these transporters in liver. The deranged folate uptake together with reduced folate transporter levels in lipid rafts resulted in reduced folate levels in liver and thereby to its reduced levels in serum of ethanol-fed rats. The chronic ethanol ingestion led to decreased folate uptake in liver, which was associated with the decreased number of transporter molecules in the lipid rafts that can be ascribed to the reduced synthesis of these transporters.

Dietary bioflavonoids represent a large class of polyphenolic compounds found in most plants. A significant number of flavonoids are reported to have beneficial health effects. Quercetin is one such flavonoid which has been reported to possess strong antioxidant properties. However, as far as male reproduction and fertility are concerned, controversial reports exist in the literature highlighting the antioxidant as well as a prooxidant character of quercetin, leaving much to the researcher's speculation. The present review therefore, aimed at addressing this paradoxical behavior of quercetin by taking into account the in vitro and in vivo studies conducted till date regarding its role in the maintenance of male reproductive potential. From the detailed survey of the published data, it appears that the conflicting biological effects of quercetin might relate to its dose and the redox state of the cell. Thus, the cellular toxicity of quercetin metabolites might overshadow the beneficial effects of its supplementation in subjects having reproductive dysfunction coupled with elevated oxidative stress leading to the paradoxical behavior of the flavonoid.

Background 14C-urea breath test (14C-UBT) is employed as a 'gold standard' technique for the detection of active gastric Helicobacter pylori infection and is recommended as the best option for "test-and-treat" strategy in primary health care centers. Aim To compare the performance of capsulated and non-capsulated 14C-UBT protocols for the detection of H. pylori infection in patients. Methods Fifty eight H. pylori infected patients underwent routine upper GI endoscopy and biopsies were processed for rapid urease test (RUT) and histopathology examination. Capsulated 14C-UBT was done in a novel way by using 74 kBq of 14C-urea along with 6.0 MBq of 99mTc-diethylene triamine penta-acetic acid (99mTc-DTPA) to simultaneously monitor the movement and the fate of ingested capsule after delineating the stomach contour by using 20.0 MBq of 99mTechnetium pertechnetate (99mTcO4-) under dual head gamma camera. Non-capsulated 14C-UBT was performed within 2 days of the previous test and the results of these protocols were compared. Results In 3 out of 58 H. pylori positive cases (5.17%), 14C-UBT results were found to be negative by using the capsulated method. Interestingly, on monitoring the real time images of the capsule in these cases it was found that misdiagnosis of H. pylori infection occurred mainly due to either rapid transit of the 14C-urea containing capsule from the upper gastric tract or its incomplete resolution in the stomach during the phase of breath collection. Conclusion Use of non-capsulated '4C-UBT protocol appears to be a superior option than the conventional capsule based technique for the detection of H. pylori infection.

Summary Oxidative stress is a leading cause of male infertility. To combat this, germ cells and spermatozoa are endowed with various enzymes, vitamins and proteins. Certain other components of food, including bioflavonoids, also provide protection against free radicals. This study analysed the effect of quercetin, a bioflavonoid, on male reproductive function in adult mice, after intraperitoneal treatment with varying concentrations of quercetin (2, 8 and 20 mg kg−1 b.wt.) for 2 weeks. Quercetin increased the generation of reactive oxygen species and lipid peroxidation in the testis with concomitant decrease in sperm count and motility in a dose-dependent manner. Activities of antioxidant enzymes catalase, superoxide dismutase and levels of reduced glutathione were found to be decreased in a dose-dependent manner. Also, the levels of oxidised glutathione were increased leading to a shift in redox ratio. The testicular histomorphology was also altered dose dependently. Germ cell kinetic study revealed significant loss of various germ cell populations with increasing dose of quercetin. Interestingly, there was a reduction in germinal epithelium thickness concomitant with an increase in seminiferous tubule lumen diameter. In conclusion, the deleterious effects of quercetin on germ cells could be attributed to its pro-oxidant ability that might affect the Sertoli cell functions.

Acute pancreatitis is an inflammatory disease of the pancreas caused by release of activated digestive enzymes in the pancreas. A number of therapeutic options have been explored for acute pancreatitis, but none has been unambiguously proven to be effective. Rosiglitazone has been shown to be efficacious in acute pancreatitis; thus, the present study was planned to evaluate the effect of rosiglitazone on pancreatic regeneration. Pancreatitis was induced by l-arginine in rats which were divided into three groups: cholecystokinin (CCK-8), rosiglitazone and vehicle. Rats were sacrificed at four time points after induction of pancreatitis i.e. 24h, day 3, day 14 and day 28 for determination of biochemical parameters and histological examination. Rate of DNA synthesis, immunohistochemistry and RT-PCR were performed at day 3 and day 7. Drug administration was started 2h after last L-arginine injection and continued till the day of sacrifice. The lower levels of enzyme in rosiglitazone-treated group compared to vehicle group proved the efficacy of rosiglitazone treatment in reducing severity of acute pancreatitis. The nucleic acid content and rate of DNA synthesis were significantly higher in rosiglitazone group indicating promotion of pancreatic regeneration. The histopathological score were lower in rosiglitazone group. Rosiglitazone treatment promoted pancreatic regeneration after acute injury. Currently, only symptomatic treatment is available, regeneration of pancreatic tissue can be a future strategy in the management of acute pancreatitis. Further studies are required to support the findings of the present study.

Background and aim: One of the active combustion product of cigarette smoke, Benzo[a]pyrenes, role in pulmonary cancer is clearly understood. However, its role in gastrointestinal cancer including colon cancer is not clearly understood. Methods: In this study, benzo(a)pyrenes was treated to colon cells to evaluate its role in cell viability, cellular ROS, and gene expression of various PLA2 isoforms was evaluated by FACS and PCR. The identified PLA2 was silenced at the gene level to evaluate its role in cell viability and ROS generation. Results: B(a)P treatment at 1 µg/mL for 48 h to HCT-15 male colon cells significantly reduced the cell viability without affecting HT-29 female colon cells. Higher doses and longer treatment duration with B(a)P showed that female colon cells were highly sensitive than male colon cells. Annexin-V/PI staining for preapoptotic detection showed that B(a)P treatment increased the apoptosis in both the cell types in a concentration and time-dependent manner. The cytosolic ROS (cROS) and superoxide radical (SOR) formation in the female colon cells was significantly higher than male colon cells unlike the mitochondrial ROS (mtROS) production which was significantly higher in male colon cells. Treatment with B(a)P significantly upregulated the IID and IVA PLA2 isoform groups in HCT-15 male colon cells, whereas IB was upregulated in HT-29 female colon cells among the various PLA2 isozyme gene studied (IB, IID, III, IVA, IVB, IVC, VI, X, aiPLA2 and iPLA2). Gene silencing experiments targeting PLA2 IID and IVA in the HCT-15 male colon cells and IB in HT-29 female colon cells showed no effect with B(a)P treatment on the cell proliferation, apoptosis, membrane integrity and free radicals (ROS, mtROS, and SOR) generation. Conclusion: Targeting specific PLA2 isozymes in a cell-specific manner abolished the B(a)P-induced PLA2-mediated oxidative damage–related signaling pathways.

Background: Cigarette smoking is responsible for various lung pathologies including chronic lung inflammation, emphysema, chronic obstructive pulmonary disease (COPD), cancer, and annually causes almost 10 million deaths globally. During smoke exposure, most affected cells are the alveolar epithelial cells where as a repair mechanism, activation of cytosolic phospholipase A2 enzymes takes place. High free radicals and cPLA2 activity due to continuous exposure of smoke exposure leads to elevated levels of secondary metabolites and various pathophysiologic conditions such as chronic inflammation, oxidative stress and cancer. To reduce the burden of chronic inflammation as well as oxidative stress, and higher levels of secondary metabolites whose role is well defined in progression of cancer, we checked the therapeutic potential of cPLA2 inhibitor arachidonyl trifluromethyl ketone (ATK) by pharmacologically targeting the most expressible cPLA2 during continuous exposure of cigarette smoke. Aim: To check the therapeutic potential of cytosolic PLA2 isoform specific inhibitor arachidonyl trifluromethyl ketone in cigarette smoke condensate–induced pathologic conditions in alveolar type I and II epithelial cells. Methods: Effect of cPLA2 inhibitor on CSC-induced cPLA2 activity were checked using colorimetric assay, cell viability using MTT assay, FDA uptake assay using fluorescence microscopy, ROS levels and apoptosis markers through flow cytometry, and ERK levels using ELISA, in both type of alveolar epithelial cells. Results: ATK significantly mimicked CSC-induced cPLA2 activity, free radicals, primary apoptosis, ratio of apoptotic/apoptotic proteins and levels of ERK whereas protected cells from loss of cell viability and membrane integrity. Conclusion: Current observations revealed cPLA2s as a potential therapeutic target and their inhibitor ATK as a potential therapeutic agent in Cigarette smoke induced pathological conditions in alveolar type I and II epithelial cells.

Helicobacter pylori (H. pylori) infection, quite prevalent in the developing countries, is considered to be one of the causative factors for various gastric pathologies and other nongastric diseases. It is believed that H. pylori infection is almost always acquired in early childhood and persists throughout life unless specific treatment is given. The (13/14)C-urea breath test (UBT) is now considered to be a 'gold standard' technique for the detection of H. pylori infection. However, because of the lack of facilities and high cost, the preferred nonradioactive ¹³C-UBT cannot be performed on pediatric patients in developing countries, whereas the radioactive ¹⁴C-UBT is not used on children because of the fear of radiation exposure. When using 37 kBq (1 μCi) of ¹⁴C-urea for the ¹⁴C-UBT, the patient is not exposed to more radiation than is acquired from the natural environment in one day, as almost all the ingested radioactivity is excreted from the body (urine and breath) within 72-120 h. This article reviews the importance of the ¹⁴C-UBT for the detection of H. pylori and justifies the radiation safety aspects of its use in children without any fear of 'radiation phobia' where the facility for ¹³C-UBT is lacking.

Introduction One of the active combustion product of cigarette smoke, Benzo[a]pyrenes, role in pulmonary cancer is clearly understood. However, its role in gastrointestinal cancer including colon cancer is not clearly understood. Material and methods In this study, benzo(a)pyrene’s was treated to colon cells to evaluate its role in cell viability, cellular ROS, and gene expression of various PLA 2 isoforms was evaluated by FACS and PCR. The identified PLA 2 was silenced at the gene level to evaluate its role in cell viability and ROS generation. Results and discussions B(a)P treatment at 1 µg/ml for 48 hour to HCT-15 male colon cells significantly reduced the cell viability without affecting HT-29 female colon cells. Higher doses and longer treatment duration with B(a)P showed that female colon cells were highly sensitive than male colon cells. Annexin-V/PI staining for pre-apoptotic detection showed that B(a)P treatment increased the apoptosis in both the cell types in a concentration and time-dependent manner. The cytosolic ROS (cROS) and superoxide radical (SOR) formation in the female colon cells was significantly higher than male colon cells unlike the mitochondrial ROS (mtROS) production which was significantly higher in male colon cells. Treatment with B(a)P significantly upregulated the IID and IVA PLA 2 isoform groups in HCT-15 male colon cells, whereas IB was upregulated in HT-29 female colon cells among the various PLA2 isozyme gene studied (IB, IID, III, IVA, IVB, IVC, VI, X, aiPLA 2 and iPLA 2 ). Gene silencing experiments targeting PLA 2 IID and IVA in the HCT-15 male colon cells and IB in HT-29 female colon cells showed no effect with B(a)P treatment on the cell proliferation, apoptosis, membrane integrity and free radicals (ROS, mtROS, and SOR) generation. Conclusion Targeting specific PLA 2 isozymes in a cell specific manner abolished the B(a)P-induced PLA 2 mediated oxidative damage related signalling pathways.

Smokeless tobacco (ST) consumption is implicated in the pathogenesis of oral diseases, including cancer. However, its pathological effect in other organs is not well understood. In the present study, the effect of aqueous extract of smokeless tobacco (AEST) prepared from "gutkha" (a form of ST) on the xenobiotic drug-metabolizing enzymes, histopathological changes, and damage to the genetic material in lung, liver, and kidney of rats was evaluated. Animals were orally administered AEST at a low dose (L-AEST, 96 mg/kg body wt/day) for 2 (L-AEST(2)) and 28 weeks (L-AEST(28)) and at a high dose (H-AEST, 960 mg/kg body wt/day) for 2 weeks (H-AEST(2)). Real-time PCR and immunohistological studies showed that administration of L-AEST(2) did not induce the expression of phase I cytochrome P450s (CYP1A1, 1A2, and 2E1) and phase II mu-glutathione-s-transferase (GST-mu) drug-metabolizing enzymes in lung, liver, and kidney. Although H-AEST(2) administration significantly induced both gene and protein expression of CYP1A1, 1A2, and 2E1 in all of the above organs, it mildly expressed the phase II detoxifying enzyme, GST-mu, in type I and type II epithelial cells of lung and in proximal tubular cells of kidney. L-AEST(28) enhanced the gene and protein expression of CYP1A1, 1A2, and 2E1 in lung, liver, and kidney in a differential manner and induced the expression of GST-mu in lung and kidney. L-AEST(28) induced the micronuclei formation in the peripheral blood mononuclear cells, TNF-alpha in plasma, and myeloperoxidase activity in the organs. L-AEST(28) significantly enhanced Bax, p53, and NF-kappaB and decreased Bcl-2 gene expressions differentially in an organ-specific manner. The differential changes in these organs due to AEST might be due to their different physiological functions and variable sensitivities toward the metabolites of AEST, which create a microenvironment favorable for AEST-induced pathogenesis. This study broadens the insight into the different molecular mechanisms in various organs, which appear to be deregulated due to AEST. Understanding these processes may help in clinical treatment planning strategies for tobacco-related diseases.

Current treatment options for acute pancreatitis are supportive and symptomatic. Due to lack of agents targeting the underlying pathophysiology a large amount of experimental work is going on to identify novel therapeutic agents. The present study was carried out to explore if melatonin can modulate the spontaneous regeneration process of the pancreas after experimentally induced acute pancreatitis. Rats were given two i.p. injections of l-arginine in a dose of 200mg/100g at an interval of 1h for induction of pancreatitis. After this rats were randomly divided into three groups i.e. saline, CCK-8 and melatonin. Drug treatment was started 2h after the last l-arginine injection and continued till the day of sacrifice. An additional only saline treated control group was included for comparison. Animals in each group were sacrificed at 24h, days 3, 14 and 28 after pancreatitis induction for determination of biochemical parameters (serum amylase, lipase and IL-10 and pancreatic amylase, total proteins and nucleic acid content) and histological examination. For rate of DNA synthesis and immunohistochemical studies animals were sacrificed at day 3 and day 7. Melatonin treatment was found to be beneficial in acute pancreatitis. Severity of acute pancreatitis was significantly reduced in melatonin group. Nucleic acid content, rate of DNA synthesis, pancreatic proteins and pancreatic amylase content were significantly improved. Histopathological examination showed significantly lower total scores in melatonin group. Results of melatonin group were comparable to that of positive control, CCK-8 group. Thus melatonin treatment was found to promote the spontaneous regeneration process of pancreatic tissue.

Progesterone exerts anti-seizure effect against several chemoconvulsants. However, there is no published report on the interaction between progesterone and picrotoxin (PTX). The present study evaluated the effects of progesterone and its active metabolite, allopregnanolone against PTX-induced seizures, brain lipid peroxidation and DNA fragmentation in male mice. Finasteride, a 5alpha-reductase inhibitor and indomethacin, an inhibitor of 3infinity-hydroxysteroid dehydrogenase were assessed against progesterone's effects on PTX-induced seizures, brain lipid peroxidation and DNA fragmentation. PTX produced clonic-tonic seizures in mice with CD50 and CD97 of 2.4 and 4.0mg/kg, i.p. respectively. Progesterone significantly countered PTX-induced seizures, with ED50 of 78.30mg/kg and ED97 of 200mg/kg. Progesterone antagonized PTX-induced DNA fragmentation. Finasteride (200mg/kg) and indomethacin (1mg/kg) reversed the anti-seizure and anti-DNA fragmentation effects of progesterone. Allopregnanolone, also protected against PTX-induced seizures and DNA fragmentation. There was no significant change in the brain lipid peroxidation parameters in any of the treatment groups. It may be concluded that progesterone protects against PTX-induced seizures and DNA fragmentation through its active metabolites allopregnanolone and 5alpha-pregnan-3,20-dione. However, it appears from the present study that, the neuroprotection with progesterone is primarily on account of allopregnalone. The therapeutic potential of allopregnanolone deserves to be evaluated clinically.

The role of oxidative stress in the pathogenesis of various conditions including epilepsy, inflammatory bowel disease and rheumatoid arthritis is evolving. The aim of this study was to find out the correlation between various inflammatory models with seizures and antioxidant parameters. Fifty-four male rats were divided into three groups of colitis, adjuvant arthritis and cotton wool granuloma (CWG). Each group had three subgroups of control, model and treatment. Thalidomide was used as treatment in colitis and arthritis group, whereas etoricoxib was used in CWG group. In colitis and arthritis groups, thalidomide was administered for 3 and 17 days, respectively, whereas etoricoxib was administered for 7 days in CWG group. At the end of treatment protocols, a subconvulsive dose of pentylenetetrazole (PTZ) (40 mg/kg i.p.) was injected intraperitoneally to note seizure onset and score. After confirming the presence of inflammation by morphological and histological studies, plasma and brain biochemical parameters of oxidative stress were estimated. The models of colitis, arthritis and CWG were effectively produced as evidenced by morphological scores (P < 0.001). Thalidomide reduced the morphological score (P < 0.002) and seizure grade (P < 0.001), whereas increased seizure onset (P < 0.001) in the arthritis group. There was an increase in malondialdehyde levels in the brain of thalidomide-treated groups (P < 0.002) and a significant decrease in superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels. There was neither improvement in seizure nor any significant changes in lipid peroxidation and antioxidant enzyme levels in etoricoxib-treated group. Thalidomide was effective in reducing the extent of arthritis as well as reducing the seizure scoring and increasing seizure onset in the adjuvant arthritis group. As it increased lipid peroxidation and reduced SOD and GPx, further evaluation is necessary with respect to oxidative stress.

Purpose To analyze p53, bcl-2, c-myc, and cyclooxygenase-2 protein expression changes and examine their relationship with various antioxidant enzymes in esophageal carcinoma patients. Methods and Materials Patients in Group 1 underwent transhiatal esophagectomy and those in Group 2 were administered chemoradiotherapy followed by surgery after 4 weeks of neoadjuvant therapy. Results The relationship analysis among the various protein markers and antioxidant enzymes showed an inverse correlation between bcl-2 and superoxide dismutase/catalase in tumor tissues, irrespective of the treatment arm followed. An important positive association was observed between bcl-2 and reduced glutathione levels in the tumor tissue of patients receiving neoadjuvant therapy. Another apoptosis-modulating marker, c-myc, in the tumor tissue of Group 2 patients showed similar pattern levels (high and low) as that of superoxide dismutase/catalase. The association of cyclooxygenase-2 and p53 with various antioxidant enzymes showed a significant positive correlation between cyclooxygenase-2 expression and catalase activity and an inverse trend between p53 expression and superoxide dismutase and catalase activity in the tumor tissue of patients given neoadjuvant therapy. In addition, patients with overexpressed p53 protein levels had lower glutathione peroxidase enzyme levels and vice versa in the tumor tissue of patients who had undergone surgery as their main mode of treatment. Conclusion The results of this study broaden the insight into the relationships shared among oncoproteins and the antioxidant defense system, and this could be helpful in the clinical management of esophageal carcinoma.

We investigated the in vitro efficacy of all-trans retinoic acid (ATRA) and alpha-tocopherol succinate (alpha-TS) alone and in combination on the induction of cell death in freshly isolated leukemic cells obtained from chronic myeloid leukemia (CML) patients. In vitro cytotoxicity and induction of lipid peroxidation by ATRA (10 microM) and alpha-TS (25 or 50 microM) were evaluated in primary leukemic cells by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and malondialdehyde formation respectively. Treatment of leukemic cells with alpha-TS alone or in combination with ATRA significantly (P < 0.05) decreased the cell viability in a concentration and time dependent manner as compared to peripheral blood mononuclear cells obtained from normal healthy controls. Lipid peroxidation was enhanced by 98% (P < 0.05) on combined treatment of cells with ATRA (10 microM) and alpha-TS (50 microM). ATRA alone did not enhance the externalization of phosphatidyl serine as studied by annexin-V binding using fluorescence activated cell sorter analysis, whereas in combination with alpha-TS it increased to 400% at 12 h. The treatment of leukemic cells to combination of ATRA with alpha-TS significantly decreased (P < 0.05) mitochondrial membrane potential and enhanced lysosomal destabilization. The combination of these drugs also increased mitochondrial and cytosolic reactive oxygen species (ROS) production, nitric oxide levels, and caspase-3 activity significantly and caused DNA fragmentation at 24 h in a concentration dependent manner in the leukemic cells. Our data suggest that ATRA in combination with alpha-TS efficiently induces apoptosis in leukemic cells, which may be a useful therapeutic modality in CML patients.

We examined the effect of whole body low-dose gamma-irradiation on the status of the antioxidant defense system in the rodent kidneys at different time intervals. Young male Balb/c mice were exposed to whole body radiation from a (60)Co source at doses of 10, 25 and 50 cGy (48.78 cGy/min). Antioxidant status and lipid peroxidation were estimated in the kidneys at 4, 12 and 24 h after irradiation. Lipid peroxidation increased between 33% and 49% and reduced glutathione between 12% and 47% at 12 h at different radiation doses. Reduced glutathione level remained significantly (p < 0.05) elevated even at 24 h after irradiation to 25 cGy. Superoxide dismutase activity also increased by 37% at 12 h on exposure of animals to all the doses up to 50 cGy. Catalase activity increased significantly at 12 h on exposure to 10 cGy and 50 cGy. Interestingly, glutathione peroxidase activity increased by 31% at 4 h and subsequently returned to control levels at 24 h after exposure to 50 cGy. Glutathione reductase activity increased by 10-12% at 12 h after exposure to 25 cGy and 50 cGy. The results suggest that the whole body exposure of animals to gamma radiation stimulates the antioxidant defense system in the kidneys within 4 to 24 h after irradiation, at doses of 25 cGy and 50 cGy.

Excessive alcohol consumption and dietary folate inadequacy are the main contributors leading to folate deficiency (FD). The present study was planned to study regulation of folate transport in conditions of FD and ethanol exposure in human embryonic kidney cell line. Also, the reversible nature of effects mediated by ethanol exposure and FD was determined by folate repletion and ethanol removal. For ethanol treatment, HEK293 cells were grown in medium containing 100 mM ethanol, and after treatment, one group of cells was shifted on medium that was free from ethanol. For FD treatment, cells were grown in folate-deficient medium followed by shifting of one group of cells on folate containing medium. FD as well as ethanol exposure resulted in an increase in folate uptake which was due to an increase in expression of folate transporters, i.e., reduced folate carrier, proton-coupled folate transporter, and folate receptor, both at the mRNA and protein level. The effects mediated by ethanol exposure and FD were reversible on removal of treatment. Promoter region methylation of folate transporters remained unaffected after FD and ethanol exposure. As far as transcription rate of folate transporters is concerned, an increase in rate of synthesis was observed in both ethanol exposure and FD conditions. Additionally, mRNA life of folate transporters was observed to be reduced by FD. An increased expression of folate transporters under ethanol exposure and FD conditions can be attributed to enhanced rate of synthesis of folate transporters.

The present study was designed to evaluate the effects of long-term use of aqueous extract of gutkha (a form of smokeless tobacco) on the antioxidant defense status and histopathological changes in liver, lung, and kidney of male Wistar rats. Animals were orally administered aqueous extract of smokeless tobacco (AEST) at a low dose (96 mg/kg body weight per day) for 2 and 32 weeks, and at a high dose (960 mg/kg body weight per day) for 2 weeks. High-dose AEST for 2 weeks decreased the hepatic glutathione (GSH) and glutathione peroxidase (GPx), and increased lipid peroxidation (Lpx) by 17%, 19%, and 20%, respectively. Low-dose AEST for 32 weeks significantly decreased (p < 0.05) the antioxidant status in these organs. In liver, AEST decreased GSH levels and the activities of superoxide dismutase (SOD), catalase (CAT), and GPx by 34.6%, 29%, 17.1%, and 17.4%, respectively, but it increased Lpx by 64%. In kidney, GSH, SOD, CAT, and GPx were decreased by 26.6%, 23%, 33%, and 18%, respectively, with an increase of Lpx by 65%. AEST decreased the lung GSH, SOD, CAT, and GPx, and increased lung Lpx by 43%, 28.5%, 37%, 40%, and 24%, respectively. However, no change in the plasma levels of vitamins A, C, and E were observed with AEST treatment. Histopathological findings suggest that administration of AEST at the high dose for 2 weeks or at the low dose for 32 weeks could cause mild to moderate inflammation in liver and lungs. In conclusion, a decrease in the antioxidant defense system and long-term inflammation caused by smokeless tobacco may be risk factors for gutkha-induced pathogenesis.

Background: 14C-urea breath test (UBT) is considered to be an accurate diagnostic test for the detection of active Helicobacter pylori infection. Various test meals are used in 14C-UBT to slow down gastric emptying, and to enhance the gastric distribution, in order to increase the time and area of contact between microorganisms and the tracer substrate. The aim of the present paper was to evaluate the effect of gastric environment on the performance of 14C-UBT using an alkaline and an acidic liquid test meal having gastric emptying retardant effect. Methods: The comparison of 14C-UBT was done with liquid test meals (200 mL water) comprising (i) plain drinking water (PDW); (ii) 1.3 g or 3.0 g citric acid (CA); and (iii) 3.0 g trisodium citrate (TSC). Eighteen patients (37 ± 12 years, range 18–57 years) with complaints of dyspepsia participated in the study. The status of H. pylori was confirmed by histology and rapid urease test. A total of 93 kBq of 14C-urea (0.5 mL) in a gelatin capsule was orally administered along with liquid test meals to the overnight fasting subjects. Breath samples were collected and radioactivity measured. Results were expressed as 14CO2/mmol exhaled CO2 as percentage of administered radioactive urea. Results: Higher acidic gastric environment (pH approx. 2.0) with CA was found to increase the exhaled 14CO2 level in a dose-dependent manner as compared to PDW and TSC meal (P

Nitric oxide is thought to be an important modulator of various functions in normal and inflamed airways. In the present study, we evaluated the effects of high vitamin E (250 mg and 1250 mg alpha-tocopheryl acetate (TA)/kg diet/10 days) on nitric oxide (NO(.)) release by alveolar macrophages (AMs) in response to lipopolysaccharide (LPS), interleukin-1beta (IL-1beta) and tumor necrosis factor (TNF-alpha). LPS and IL-1beta treatment (1-10 microg/ml) enhanced NO(.) release in AMs from control animals fed on 50 mg vitamin E/kg diet in a concentration dependent manner. However, this enhancement of NO(.) was attenuated in the AMs of animals fed with 250 mg or 1250 mg vitamin E/kg diet. TNF-alpha had no effect in eliciting the release of NO(.) in AMs obtained either from control or from hyper vitamin E fed animals. Further, LPS (1-10 microg/ml) enhanced the inducible nitric oxide synthase (iNOS) activity of AMs of control group and TA-fed animals almost to equal extent. Similarly, LPS-induced formation of N-nitrosamine (N-nitroso-L-[(14)C]-proline) in AMs of control and TA-supplemented animals were not different statistically. On the other hand, in vitro addition of vitamin E (200 microM) in AMs of control animals, when triggered with 10 microg LPS/ml, caused a significant decrease in N-nitroso-L-[(14)C]-proline formation. It seems that high doses of TA in diet may play a role in reducing the lipopolysaccharide and proinflammatory cytokines-induced NO(.)-mediated damage by AMs.

To evaluate the effect of low-dose (50 cGy) whole body ?-irradiation on the antioxidant defense system in the liver and the lungs of mice at various post-irradiation intervals.Male Balb/c mice, 5 - 6 weeks of age, were divided into irradiated and non-irradiated groups. Whole body irradiation was done with gamma-rays from a (60)Co source at doses of 10, 25 and 50 cGy (48.78 cGy/min). Lipid peroxidation and antioxidant status were measured in the liver and the lungs at 4, 12 and 24 h after irradiation.Lipid peroxidation increased by 1.38 and 2.0 fold in lung and liver respectively at 12 h after exposure to 25 cGy. Whole body exposure to 25 and 50 cGy significantly (p0.05) increased the hepatic reduced glutathione at 4 h. Reduced glutathione continued to rise until 12 h and returned to the basal level at 24 h, whereas in the lungs it remained elevated until 24 h at 10 and 25 cGy. Antioxidant enzymes activities for superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase increased by 1.22, 1.13, 1.22 and 1.11 fold respectively (p0.05) in the liver at 4 h after exposure to 50 cGy and remained elevated at almost the same level up to 12 h after exposure. Surprisingly these antioxidant defense enzymes remained unaltered in the lung at the above radiation doses.Low-dose whole body gamma-irradiation differentially modulates the antioxidant defense system in the liver and lungs of mice. The induction of endogenous glutathione, immediately after exposure to low-dose -irradiation, may be beneficial in protecting the cells from reactive oxygen species (ROS) induced oxidative stress.

Increased levels or overexpression of ornithine decarboxylase (ODC), a rate-limiting enzyme in polyamine biosynthesis pathway, is characteristic of tumor cells. Similarly, prostaglandins (PGs) appear to be important in the pathogenesis of cancer because these affect mitogenesis, cellular adhesion, immune surveillance and apoptosis. Cancers form much more PGs than the original tissue from which they have arisen. This study has revealed that pretreatment of mice with resveratrol at a dose of 2.5 mg/kg body weight for two weeks blocked the N-nitrosodiethylamine(NDEA)-induced cytosolic ODC levels in the liver and lungs. The blockage was pronounced in hepatic tissue compared to pulmonary tissue. Resveratrol feeding caused a significant reduction in microsomal cyclooxygenase (COX) activities in the liver and lungs, while the dosage of NDEA (200 mg/kg body weight) induced COX activity 24 h after its administration. In any case, resveratrol pretreatment turned out to effectively block the induction of COX activity in the lungs by NDEA.

Human sperm motility, viability and lipid peroxidation were assessed in Ringer-Tyrode supplemented with different concentrations of quercetin. An irreversible and dose-dependent fall in sperm motility was observed with quercetin (5-200 microm). However, sperm viability was decreased only at higher concentrations (50-100 microm) of quercetin. This inhibition in sperm motility and viability has been linked with decreased Ca2+-ATPase activity, as there was no effect of quercetin on sperm lipid peroxidation.

Naturally occurring plant polyphenols, which include ellagic acid (EA), tannic acid (TA), caffeic acid (CA), and ferulic acid (FA), were tested for their superoxide anion radical (SOR)-scavenging activities. SOR were produced by interaction of the tumor promoter benzoyl peroxide (BPO) with murine peritoneal macrophages in vitro. The levels of SOR were assessed microscopically by counting the number of formazan-positive cells per 250 cells produced by the reduction of nitro blue tetrazolium. BPO at a concentration of 15 micrograms/1.85 x 10(6) cells/0.5 ml induced maximum formation of SOR in resident and thioglycollate-elicited cells. All the tested polyphenols were able to inhibit the formation of SOR induced by the tumor promoter to a variable degree. Inhibition of BPO-induced SOR formation by polyphenols was in the following order: FATACAEA. BPO stimulated the accumulation of diacylglycerol (DAG) in resident and elicited macrophages with concurrent release of choline equivalents from macrophages. Polyphenols inhibited DAG accumulation, which paralleled the inhibition of choline equivalent release. FA was observed to be the most effective and EA the least effective inhibitor of SOR formation, DAG accumulation, and release of choline equivalents. It is likely that inhibition of SOR formation might be due to some interference in the cellular lipid metabolism and phospholipid equivalent deacylation and choline release.

Over the last decade, although investigations have suggested that vitamin E affects the immune response, not much is known about its affect on the alveolar macrophage functions. In the present study, we have investigated the effect of high vitamin E (DL-alpha-tocopheryl acetate, alpha-TA) supplementation for 10 d on the activation state of rat alveolar macrophages induced by lipopolysaccharide (LPS), interleukin (IL)-1beta or tumor necrosis factor (TNF)-alpha on the basis of their ability to produce reactive oxygen species (ROS), such as superoxide (O2-*) and H2O2. LPS treatment (1 and 10 microg/mL) caused 2.44 and 2.54-fold increases in O2-*, and 2.1 and 2.3-fold increases in H2O2, respectively, from alveolar macrophages (AMs) in the diet group fed 50 mg alpha-TA/kg. However, this enhancement was not observed for the AMs of the diet groups fed 250 or 1,250 mg alpha-TA/kg. Similar results were obtained on treating the AMs with proinflammatory cytokines IL-1beta or TNF-alpha. The observed suppression in ROS release in response to various stimulants may be due to the direct and/or indirect effect of high vitamin E (250 and 1,250 mg alpha-TA/kg diet) supplementation. It may therefore, be concluded that high alpha-TA supplementation in the diet modulates the activation of AMs in rats.

For the past decade there has been emphasis on the immunomodulatory effects of vitamin E apart from its established role as a free radical scavenger. In alveolar macrophages (AMs), the role of vitamin E supplementation has not yet been investigated sufficiently. In the present study we have evaluated the effects of high vitamin E (DL-alpha-tocopheryl acetate, alpha-TA) supplementation for 10 d on rat-alveolar macrophageal antioxidant defense and reactive oxygen species (ROS) release. There was an increase in plasma vitamin E content from 5.22 +/- 1.30, at 50 mg to 12.23 +/- 1.06 and 22.32 +/- 2.02 micrograms/mL at 250 and 1,250 mg alpha-TA/kg dietary supplementation. Alveolar macrophage-vitamin E content enhanced by 56 to 75% at 250 and 1,250 mg alpha-TA diet as compared with control diet. Superoxide dismutase activity decreased and catalase and glutathione peroxidase activities increased significantly in AMs of 250 and 1,250 mg alpha-TA diet-fed rats. Reduced glutathione, total glutathione, and glutathione-S-transferase activity in AMs did not change significantly at any of the high doses of alpha-TA supplementation. On stimulation of AMs by phorbol-12-myristate-13-acetate (PMA), there was 2.8- and 3.5-fold enhancement in superoxide (O2-.) and H2O2 production, respectively, at 50 mg alpha-TA dose. But this increase by PMA could not take place in AMs from animals supplemented with 250 and 1,250 mg alpha-TA. It may therefore be concluded that high alpha-TA supplementation in diet may equip the AMs with a stronger defense against oxygen-free radical mediated damage to them.

Ellagic acid (EA), tannic acid (TA), caffeic acid (CA), and ferulic acid (FA) offer considerable promise as anticarcinogens. The role of these dietary polyphenols was investigated in the promotional phase of carcinogenesis. Topical application of polyphenols simultaneously with phorbol-12-myristate-13-acetate (PMA) or mezerein resulted in significant protection against 7,12-dimethyl-benz[a]anthracene-induced skin tumors in mice. Caffeic acid was the most effective inhibitor of tumor promotion. In vivo and in vitro treatment of murine peritoneal macrophages with the tumor promoters resulted in stimulation of superoxide anion radical formation. Tannic acid, caffeic acid, and ferulic acid were stronger inhibitors of PMA- and mezerein-induced superoxide anion radical than ellagic acid in in vivo and in vitro conditions. Treatment of [1(3)-14C]glycerol- or [methyl-14C]choline chloride-labeled resident or thioglycollate-elicited macrophages with PMA and mezerein led to accumulation of radioactive diacylglycerol equivalents. The polyphenols were capable of inhibiting these releases.

The effect of non-steroidal anti-inflammatory drugs (NSAIDs) diclofenac and naproxen on xenobiotic-metabolizing enzymes was studied in male Swiss mice given the drugs in their diet. Diclofenac at levels of 25, 75, and 375ppm, and naproxen at 150, 500, and 1, 500ppm, were given for a period of 4 weeks. Control animals consumed a similar diet deprived of the test NSAIDs. Feeding of NSAIDs did not affect the activity of arylhydrocarbon hydroxylase, whereas the level of cytochrome P-450 (P-450) was significantly decreased in liver and lungs of the animals. The maximum decline in content of the enzyme was found at the largest dose of NSAIDs. On the other hand, the activity of glutathione-S-transferase (GST) was significantly increased in the two organs by both drugs. However, hepatic activity of the enzyme after induction by 375ppm diclofenac was almost 1.37 fold greater in comparison to that after induction by 1, 500ppm naproxen. In lungs, this ratio was found to be 1.76. Like GST, reduced glutathione (GSH) levels in the liver and lungs of the animals were also significantly increased. The maximum increase in GSH in lungs elicited by the two NSAIDs was similar. The results of this study indicate that diclofenac might prove to be a better chemopreventive agent against xenobiotics because of the higher induction of GST activity by it.

Feeding of gallic acid elevated the hepatic levels of glutathione peroxidase (GPx) and catalase, whereas NADPH-mediated microsomal lipid peroxidation was found to be unaltered. However, in lungs, besides a significant increase in reduced glutathione (GSH) and GPx, there was an inhibition of enzymatic lipid peroxidation in post-mitochondrial supernatant. On the other hand, propylgallate increased the hepatic GPx and GSH as well as inhibited hepatic microsomal and pulmonary lipid peroxidation. It seems that propylgallate is a better antioxidant than gallic acid since unlike gallic acid it protects both liver and lungs from the damaging action of the free radical-mediated process of lipid peroxidation.

Alveolar macrophages (AMs) may contribute to inflammation in multiple ways, including the release of reactive oxygen species (ROS) such as superoxide anion (O2-) and hydrogen peroxide (H2O2). This provides a basis for a plausible and testable hypothesis by which inflammation and the disease might be related; that is, the levels of ROS generated by inflammatory phagocytes might be dependent upon the type of disease. Alveolar macrophages were prepared from 25 lung cancer patients, 58.7±7.51 years of age (mean±SD). Alveolar macrophages from 12 patients, 45.3±14.6 years of age, with nonmalignant lung diseases were also studied. Production of oxygen radical species was higher in AMs from the malignant lobe of lung cancer patients than in those from the disease-free lobe. However, the levels in AMs from the disease-free lobe were comparable to the levels in AMs from patients with nonmalignant lung diseases. There was an increase in hydrogen peroxide formation in the cells from malignant lobe of lung cancer patients compared with that in those from the disease-free lobe (38.4±4.16 vs. 26.9±2.94nmol/mg). The formation in the cells from patients with nonmalignant lung diseases was found to be 18.0±1.19nmol/mg protein. In the presence of phorbol-12-myristate-13-acetate, the formation in the cells from malignant lobe of lung cancer patients increased compared with that from the disease-free lobe (63.9±7.65 vs. 34.3±4.95nmol/mg protein), and the formation in the cells from patients with nonmalignant lung diseases was 42.5±5.03nmol/mg protein. The enormous increase in O2- and H2O2 production in malignancy needs further investigation for its diagnostic and prognostic values.

Background and aims (14) C-urea breath test ((14) C-UBT) is considered as "gold standard" for detection of active gastric H. pylori infection. However, till date no comparative study using encapsulated and non-encapsulated (14) C-UBT protocols has been conducted in same subjects in identical conditions. We monitored gastric fate of capsule containing (14) C-urea with real time display and compared sensitivities of these protocols at different time points of breath collection. Methods Non-encapsulated (14) C-UBT was performed using 74 kBq of (14) C-urea in 100 dyspeptic patients by collecting breath samples at 10, 15 and 20 minutes. Thereafter, within 2 days a gelatin capsule containing (14) C-urea along with 6.0 MBq of (99m) Tc-diethylene triamine penta-acetic acid was administered to each patient for real time display of capsule movement and its fate in gastrointestinal tract by gamma camera. Simultaneously, breath samples were collected for (14) CO2 measurement during image acquisition. Results Employing non-encapsulated (14) C-UBT, 74 out of 100 dyspeptic patients were found to be H. pylori positive. Discordant (14) C-UBT results were obtained in 4/74 (5.4%) cases using these two protocols. By employing encapsulated and nonencapsulated (14) C-UBT protocols, sensitivities of (14) C-UBT were found to be 90.5 versus 98.6% at 10 and 91.8 versus 97.2% at 15 minutes respectively; while these were 94.6 versus 100, 90.7 versus 98.6 and 83.7 versus 93.2% considering any one, two or all three positive values respectively. Conclusions Incomplete/non-resolution of (14) C-urea capsule in stomach during the phase of breath collections appears to decrease sensitivity of encapsulated (14) C-UBT as compared to nonencapsulated protocol for detection of H. pylori infection.

The present study demonstrates that curcumin acts as pro-oxidant and sensitizes human lung adenocarcinoma epithelial cells (A549) to apoptosis via intracellular redox status mediated pathway. Results indicated that curcumin induced cell toxicity (light microscopy and MTT assay) and apoptosis (AnnexinV-FITC/PI labeling and caspase-3 activity) in these cells. These events seem to be mediated through generation of reactive oxygen species (ROS) and superoxide radicals (SOR) and enhanced levels of lipid peroxidation. These changes were accompanied by increase in oxidized glutathione (GSSG), reduced glutathione (GSH) and gamma-glutamylcysteine synthetase (gamma-GCS) activity, but decrease in GSH/GSSG ratio. The induction of apoptosis and decrease in GSH/GSSG ratio was also accompanied by sustained phosphorylation and activation of p38 mitogen activated protein kinase (MAPK). On the other hand, addition of N-acetyl cysteine (NAC), an antioxidant, blocked the curcumin-induced ROS production and rescued malignant cells from curcumin-induced apoptosis through caspase-3 deactivation. However, L-buthionine sulfoximine (BSO), a GSH synthesis blocking agent, further enhanced curcumin-induced ROS production and apoptosis in A549 cells. Decreased GSH/GSSG ratio seems to be a crucial factor for the activation of MAPK signaling cascade by curcumin. The study therefore, provides an insight into the molecular mechanism involved in sensitization of lung adenocarcinoma cells to apoptosis by curcumin.

Summary Resveratrol has been considered as an antioxidant in biological system. However, its role in male reproductive biology has not yet been evaluated in much detail. Present study analysed its effect on male reproductive function in adult mouse, after its intraperitoneal administration (2, 8 and 20 mg kg b.wt.−1 per day) for 2 weeks. The generation of reactive oxygen species and lipid peroxidation in testis increased with concomitant decrease in sperm count and motility following resveratrol treatment as compared with the control group. Resveratrol had a negative impact on the activities of antioxidant enzymes namely catalase and superoxide dismutase and on the level of reduced glutathione. Increase in the level of oxidised glutathione by resveratrol lead to a shift in the redox ratio. Additionally, a significant loss of Leydig cells and alterations in testicular histomorphology (excessive vacuolisation and shrinkage of seminiferous tubules) was also observed. In conclusion, the deleterious effects of resveratrol on germ cells could be attributed to its pro-oxidant ability leading to testicular tissue damage.

Naturally occurring plant phenols, which included ellagic acid (EA), tannic acid (TA), caffeic acid (CA), and ferulic acid (FA), were tested for their superoxide anion radical (SOR)-scavenging activities. SOR was produced by interaction of tumor promoters, i.e. phorbol-12-myristate-13-acetate (PMA) or mezerein, in vitro with murine peritoneal macrophages. The levels of SOR were assessed microscopically by counting the number of formazan-positive (F+) cells/250 cells produced due to the reduction of nitroblue-tetrazolium. PMA at a concentration of 0.1μg/1.85×106cells/0.5ml induced maximum formation of SOR both in resident and thioglycollate-elicited cells, whereas mezerein induced the maximum formation of SOR at 10μg/1.85×106cells/0.5ml. All the tested polyphenols were able to inhibit the formation of SOR induced by the tumor promoters to a variable degree. The maximum inhibition by EA of SOR formation induced by PMA or mezerein was at a concentration of 250μM or 125μM, respectively. The polyphenols CA, TA, and FA were more effective in inhibition of SOR stimulation at a concentration of 120 and 60μM with promoters PMA and mezerein, respectively. However, ferulic acid was found to be the most effective inhibitor of SOR formation.

Dietary administration of phenolics, gallic acid, and its synthetic analogue propylgallate at varying dose levels for the period of 4 and 8 weeks to inbred male Swiss mice resulted in a significant increase in hepatic cytochrome P-450 (Cyt P-450) without any alteration in arylhydrocarbon hydroxylase (AHH) and cytochrome b5 (Cyt b5) activities. The activity of glutathione-S-transferase (GST) showed a significant elevation by these phenolics in both liver and lungs. However, they had a differential effect on the levels of reduced glutathione (GSH) in both organs. Increased levels of GST and GSH elicited by these phenolics may protect the organism from the insult of various toxic carcinogenic chemicals.

Dietary bioflavonoids represent a large class of polyphenolic compounds found in most plants. A significant number of flavonoids are reported to have beneficial health effects. Quercetin is one such flavonoid which has been reported to possess strong antioxidant properties. However, as far as male reproduction and fertility are concerned, controversial reports exist in the literature highlighting the antioxidant as well as a prooxidant character of quercetin, leaving much to the researcher's speculation. The present review therefore, aimed at addressing this paradoxical behavior of quercetin by taking into account the in vitro and in vivo studies conducted till date regarding its role in the maintenance of male reproductive potential. From the detailed survey of the published data, it appears that the conflicting biological effects of quercetin might relate to its dose and the redox state of the cell. Thus, the cellular toxicity of quercetin metabolites might overshadow the beneficial effects of its supplementation in subjects having reproductive dysfunction coupled with elevated oxidative stress leading to the paradoxical behavior of the flavonoid.

Folate mediated one-carbon metabolism is of fundamental importance for various cellular processes, including DNA synthesis and methylation of biological molecules. Due to the exogenous requirement of folate in mammals, there exists a well developed epithelial folate transport system for regulation of normal folate homeostasis. The intestinal and renal folate uptake is tightly and diversely regulated and disturbances in folate homeostasis like in alcoholism have pathological consequences. The study was sought to delineate the regulatory mechanism of folate uptake in intestine and reabsorption in renal tubular cells that could evaluate insights of malabsorption during alcoholism. The folate transporters PCFT and RFC were found to be associated with lipid rafts of membrane surfaces in intestine and kidney. Importantly, the observed lower intestinal and renal folate uptake was associated with decreased levels of folate transporter viz. PCFT and RFC in lipid rafts of intestinal and renal membrane surfaces. The decreased association of folate transporters in lipid rafts was associated with decreased protein and mRNA levels. In addition, immunohistochemical studies showed that alcoholic conditions deranged that localization of PCFT and RFC. These findings could explain the possible mechanistic insights that may result in folate malabsorption during alcoholism.

Folates, an essential component (important B vitamin) in the human diet, are involved in many metabolic pathways, mainly in carbon transfer reactions such as purine and pyrimidine biosynthesis and amino acid interconversions. Deficiency of this micronutrient leads to the disruption of folate-dependent metabolic pathways that lead to the development of clinical abnormalities ranging from anaemia to growth retardation. Folate deficiency due to alcohol ingestion is quite common, primarily due to malabsorption. The present study dealt with the mechanistic insights of folate malabsorption in colonic basolateral membrane (BLM). Wistar rats (n12) were fed 1 g/kg body weight per d ethanol (20 %) solution orally for 3 months and folate transport was studied in the isolated colonic BLM. The folate exit across colon BLM shows characteristics of carrier-mediated process with the major involvement of reduced folate carrier (RFC). The chronic ethanol ingestion decreased the uptake by decreasing the affinity by 46 % (P P P P P P

Scopolamine, an anticholinergic, is an attractive amnesic agent for discerning the action of candidate antiamnesic drugs.Bacopa monnieraLinn (Syn. Brahmi) is one such antiamnesic agent that is frequently used in the ancient Indian medical system. We have earlier reported the reversal of diazepam-induced amnesia withB. monniera.In this study we wanted to test if scopolamine-induced impairment of spatial memory can also be ameliorated byB. monnierausing water maze mouse model. The objective of study was to study the effect ofB. monnieraon scopolamine-induced amnesia. We employed Morris water maze scale to test the amnesic effect of scopolamine and its reversal byB. monniera.Rotarod test was conducted to screen muscle coordination activity of mice. Scopolamine significantly impaired the acquisition and retrieval of memory producing both anterograde and retrograde amnesia.Bacopa monnieraextract was able to reverse both anterograde and retrograde amnesia. We propose thatB. monniera'seffects on cholinergic system may be helpful for developing alternative therapeutic approaches for the treatment of Alzheimer's disease.

The possible mechanisms of inhibition of benzo(a)pyrene (BaP) binding to DNA in vivo and in vitro by ellagic acid (EA) were investigated. In the in vitro studies BaP was activated by cytochrome P-450-dependent mixed function oxidase system, ascorbate-dependent lipid peroxidation, and peroxidation of linoleic acid. Reactions were performed in the dark at 37°C for 20 to 30min in buffered aqueous solutions with 2mg DNA and 40nmol tritiated BaP. The levels of BaP-DNA adducts were determined by assay of the radioactivity. BaP-DNA adduct levels ranged from 146 to 170fmol/mg DNA in ascorbate-and NADPH-dependent reactions and the level was 8.25±0.425pmol/mg DNA in the hematin-mediated linoleic acid reaction. Addition of EA to the reaction mixtures resulted in a significant inhibition in BaP-DNA adduct formation, depending upon the concentration of EA; e.g., 500μM EA resulted in 42 to 53% inhibition of binding in the three systems of carcinogen activation. Similarly, EA feeding to male NMRI mice at a concentration of 12μg/ml drinking water significantly decreased the levels of carcinogen-DNA adducts in the lungs. Reactions performed in vitro, in which DNA was preincubated overnight at 4°C with EA, revealed a decrease of 20% in adduct formation. MDA formation during the process of lipid peroxidation, stimulated by NADPH and ascorbate in the liver microsomes of mouse and by hematin in linoleic acid, was taken as an index for the free radical reactions. Addition of EA to in vitro systems inhibited the MDA formation with an IC50 (concentration for 50% inhibition) of 480, 400, and 400μM for NADPH, ascorbate, and hematin-mediated free radical reactions, respectively.

to the editor: The article recently published by Kim and coworkers ([3][1]) is very interesting and presents a new modality for the prevention of ischemia-reperfusion (I/R) injury in the mouse kidney. The authors have reported that whole body irradiation preconditioning (IP) of the animals with an

The role of oxidative stress in the pathogenesis of various conditions including epilepsy, inflammatory bowel disease and rheumatoid arthritis is evolving. The aim of this study was to find out the correlation between various inflammatory models with seizures and antioxidant parameters. Fifty-four male rats were divided into three groups of colitis, adjuvant arthritis and cotton wool granuloma (CWG). Each group had three subgroups of control, model and treatment. Thalidomide was used as treatment in colitis and arthritis group, whereas etoricoxib was used in CWG group. In colitis and arthritis groups, thalidomide was administered for 3 and 17 days, respectively, whereas etoricoxib was administered for 7 days in CWG group. At the end of treatment protocols, a subconvulsive dose of pentylenetetrazole (PTZ) (40 mg/kg i.p.) was injected intraperitoneally to note seizure onset and score. After confirming the presence of inflammation by morphological and histological studies, plasma and brain biochemical parameters of oxidative stress were estimated. The models of colitis, arthritis and CWG were effectively produced as evidenced by morphological scores (P0.001). Thalidomide reduced the morphological score (P0.002) and seizure grade (P0.001), whereas increased seizure onset (P0.001) in the arthritis group. There was an increase in malondialdehyde levels in the brain of thalidomide-treated groups (P0.002) and a significant decrease in superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels. There was neither improvement in seizure nor any significant changes in lipid peroxidation and antioxidant enzyme levels in etoricoxib-treated group. Thalidomide was effective in reducing the extent of arthritis as well as reducing the seizure scoring and increasing seizure onset in the adjuvant arthritis group. As it increased lipid peroxidation and reduced SOD and GPx, further evaluation is necessary with respect to oxidative stress.