Biomedical subjects

Manoj K Pandey

Publications and source records attributed to Manoj K Pandey.

11 recordsLinked to original sources

PubMed2007

Celastrol, a novel triterpene, potentiates TNF-induced apoptosis and suppresses invasion of tumor cells by inhibiting NF-kappaB-regulated gene products and TAK1-mediated NF-kappaB activation.

Celastrol, a quinone methide triterpene derived from the medicinal plant Tripterygium wilfordii, has been used to treat chronic inflammatory and autoimmune diseases, but its mechanism is not well understood. Therefore, we investigated the effects of celastrol on cellular responses activated by TNF, a potent proinflammatory cytokine. Celastrol potentiated the apoptosis induced by TNF and chemotherapeutic agents and inhibited invasion, both regulated by NF-kappaB activation. We found that TNF induced the expression of gene products involved in antiapoptosis (IAP1, IAP2, Bcl-2, Bcl-XL, c-FLIP, and survivin), proliferation (cyclin D1 and COX-2), invasion (MMP-9), and angiogenesis (VEGF) and that celastrol treatment suppressed their expression. Because these gene products are regulated by NF-kappaB, we postulated that celastrol mediates its effects by modulating the NF-kappaB pathway. We found that celastrol suppressed both inducible and constitutive NF-kappaB activation. Celastrol was found to inhibit the TNF-induced activation of IkappaBalpha kinase, IkappaBalpha phosphorylation, IkappaBalpha degradation, p65 nuclear translocation and phosphorylation, and NF-kappaB-mediated reporter gene expression. Recent studies indicate that TNF-induced IKK activation requires activation of TAK1, and we indeed found that celastrol inhibited the TAK1-induced NF-kappaB activation. Overall, our results suggest that celastrol potentiates TNF-induced apoptosis and inhibits invasion through suppression of the NF-kappaB pathway.

Active Transport, Cell Nucleus↗

PubMed2006

Poly-(3-hexylthiophene) self-assembled monolayer based cholesterol biosensor using surface plasmon resonance technique.

Cholesterol oxidase (ChOx) has been covalently immobilized onto 1-fluoro-2-nitro-4-azidobenzene (FNAB) modified poly-(3-hexylthiophene) (P3HT) self-assembled monolayer (SAM) onto gold coated glass plates. These ChOx/FNAB/P3HT/Au bio-electrodes have been characterized using contact angle (CA) measurements, UV-vis spectroscopy, electrochemical impedance technique, cyclic voltammetric technique and atomic force microscopic (AFM) technique, respectively. The ChOx/FNAB/P3HT/Au bio-electrodes were utilized for the estimation of cholesterol concentration in standard solutions using surface plasmon resonance (SPR) technique. It is shown that this SPR biosensor has linearity from 50 to 500 mg/dl of cholesterol in solution with detection limit of 50 mg/dl, sensitivity of 1.0 4 m degrees /(mg dl), reusability of around 15 times and a shelf-life of about 10 weeks when stored at 4 degrees C.

Adsorption↗

PubMed2006

Inflammation and cancer: how hot is the link?

Although inflammation has long been known as a localized protective reaction of tissue to irritation, injury, or infection, characterized by pain, redness, swelling, and sometimes loss of function, there has been a new realization about its role in a wide variety of diseases, including cancer. While acute inflammation is a part of the defense response, chronic inflammation can lead to cancer, diabetes, cardiovascular, pulmonary, and neurological diseases. Several pro-inflammatory gene products have been identified that mediate a critical role in suppression of apoptosis, proliferation, angiogenesis, invasion, and metastasis. Among these gene products are TNF and members of its superfamily, IL-1alpha, IL-1beta, IL-6, IL-8, IL-18, chemokines, MMP-9, VEGF, COX-2, and 5-LOX. The expression of all these genes are mainly regulated by the transcription factor NF-kappaB, which is constitutively active in most tumors and is induced by carcinogens (such as cigarette smoke), tumor promoters, carcinogenic viral proteins (HIV-tat, HIV-nef, HIV-vpr, KHSV, EBV-LMP1, HTLV1-tax, HPV, HCV, and HBV), chemotherapeutic agents, and gamma-irradiation. These observations imply that anti-inflammatory agents that suppress NF-kappaB or NF-kappaB-regulated products should have a potential in both the prevention and treatment of cancer. The current review describes in detail the critical link between inflammation and cancer.

Anti-Inflammatory Agents↗

PubMed2006

Induction of P53, P21Waf1, orinithine decorboxylase activity, and DNA damage leading to cell-cycle arrest and apoptosis following topical application of repeated fish fried oil extract to mice.

Repeated frying of food produces numerous carcinogens including polycyclic aromatic hydrocarbons (PAHs). Our prior studies have shown that repeated fish fried oil extract (RFFE) induces cytochrome P (CYP)-450 1A1/2 isozymes thereby causing increased generation of electrophilic reactive metabolites of PAHs and subsequent binding to DNA. In the present study, molecular events associated with DNA damage, apoptosis, and proliferation following topical exposure to RFFE have been investigated in mice. Single topical application of RFFE (500 microg) for 24-48 h caused significant DNA damage with Comet assay in terms of olive tail moment (OTM) (204-246%), tail DNA (253-293%), and tail length (172-195%). Overexpression of p53 and p21WAF1 proteins was observed in skin cells following single topical exposure of RFFE for 24-72 h, which was similar to that of benzo(a)pyrene (BP) exposure (24 h). Though RFFE and BP exposure separately, did not result in G(0)/G(1) arrest, but a significant increase in the proportion of cells in S-phase was observed. Apoptotic induction was noticed in skin cells, with maximum induction after 48 h of exposure to RFFE. Further, topical treatment of mice with RFFE (500 microg) for 6 h significantly increased orinithine decarboxylase (ODC) activity by 7.5-fold when compared to control. These results indicate that RFFE exposure caused ODC induction accompanied by increased levels of p53 and p21WAF1 proteins leading of apoptosis and delay of cells in S-phase thereby indicating the possible carcinogenic potential of RFFE.

Administration, Topical↗

PubMed2006

Assessment of carcinogenic potential of repeated fish fried oil in mice.

Our prior studies have shown that single topical treatment of repeated fish fried oil extract (RFFE), containing various polycyclic aromatic hydrocarbons (PAHs), to the dorsal epidermis of mice caused enhancement of DNA damage along with higher expression of p53 and p21WAF1 proteins and cell-cycle arrest. In the present study carcinogenic potential of repeated fish fried oil (RFFO) and RFFE was assessed. Single topical application of RFFO (100 microL/animal) and RFFE (100-500 microg/animal) to Swiss albino female mice resulted in significant induction (1.8- to 7.4-fold) of ornithine decarboxylase activity. Twice weekly topical application of methylcholanthrene (MCA) for 24 wk or single topical application of 7,12-dimethylbenzanthracene (DMBA) or RFFO or RFFE, as initiator followed by twice weekly application of 12-O-tetradecanoyl phorbol myristate acetate (TPA) as promoter for 24 wk, resulted in development of skin papillomas after 6, 7, 18, and 9 wk, respectively. The cumulative number of tumors in MCA, DMBA/TPA, RFFE (200 microg)/TPA, and RFFE (500 microg)/TPA groups were 276, 168, 34, and 58 after 24 wk while negligible or minimal initiating activity was noticed in RFFO/TPA group. No tumors were found in animals either given twice weekly topical application of RFFO or a single initiating dose of DMBA followed by twice weekly application of RFFO. Histopathology of skin of animals treated with RFFE/TPA showed marked proliferation of epidermal layers along with abnormal mitosis and multinucleated tumor appearance. Skin of animals in groups RFFO/TPA and DMBA/RFFO showed sloughing and regeneration of epidermal layers, oedema along with proliferation of fibroblasts. Histochemical localization of gamma-glutamyl transpeptidase was found to be substantially higher in skin of mice treated with RFFO/TPA and RFFE/TPA. Animals treated with RFFO/TPA, DMBA/RFFO, and RFFE/TPA resulted in significant induction of cutaneous aryl hydrocarbon hydroxylase (AHH) (421-432%), ethoxyresorufin-O-deethylase (252-316%), and glutathione S-transferase (133-245%) activities. Animals treated with RFFO/TPA, DMBA/RFFO, and RFFE/TPA led to significant reduction in glutathione content (39-44%) with a concomitant increase in lipid peroxidation (254-492%). Animals treated with RFFO/TPA and RFFE/TPA led a significant decrease in catalase (43-69%) and superoxide dismutase (20-31%) activities while glutathione reductase activity was found to be diminished (23-51%) in RFFO, RFFO/TPA, DMBA/RFFO, and RFFE/TPA treated groups. These results suggest that RFFE possess skin tumor initiating activity and that it may have weak promoting activity as well, which may involve free radicals.

Administration, Topical↗

PubMed2006

A regulatory role for the C5a anaphylatoxin in type 2 immunity in asthma.

Complement component 5 (C5) has been described as either promoting or protecting against airway hyperresponsiveness (AHR) in experimental allergic asthma, suggesting pleomorphic effects of C5. Here we report that local pharmacological targeting of the C5a receptor (C5aR) prior to initial allergen sensitization in murine models of inhalation tolerance or allergic asthma resulted in either induction or marked enhancement of Th2-polarized immune responses, airway inflammation, and AHR. Importantly, C5aR-deficient mice exhibited a similar, increased allergic phenotype. Pulmonary allergen exposure in C5aR-targeted mice resulted in increased sensitization and accumulation of CD4+ CD69+ T cells associated with a marked increase in pulmonary myeloid, but not plasmacytoid, DC numbers. Pulmonary DCs from C5aR-targeted mice produced large amounts of CC chemokine ligand 17 (CCL17) and CCL22 ex vivo, suggesting a negative impact of C5aR signaling on pulmonary homing of Th2 cells. In contrast, C5aR targeting in sensitized mice led to suppressed airway inflammation and AHR but was still associated with enhanced production of Th2 effector cytokines. These data suggest a dual role for C5a in allergic asthma, i.e., protection from the development of maladaptive type 2 immune responses during allergen sensitization at the DC/T cell interface but enhancement of airway inflammation and AHR in an established inflammatory environment.

Allergens↗

PubMed2006

From traditional Ayurvedic medicine to modern medicine: identification of therapeutic targets for suppression of inflammation and cancer.

Cancer is a hyperproliferative disorder that involves transformation, dysregulation of apoptosis, proliferation, invasion, angiogenesis and metastasis. Extensive research during the last 30 years has revealed much about the biology of cancer. Drugs used to treat most cancers are those that can block cell signalling, including growth factor signalling (e.g., epidermal growth factor); prostaglandin production (e.g., COX-2); inflammation (e.g., inflammatory cytokines: NF-kappaB, TNF, IL-1, IL-6, chemokines); drug resistance gene products (e.g., multi-drug resistance); cell cycle proteins (e.g., cyclin D1 and cyclin E); angiogenesis (e.g., vascular endothelial growth factor); invasion (e.g., matrix metalloproteinases); antiapoptosis (e.g., bcl-2, bcl-X(L), XIAP, survivin, FLIP); and cellular proliferation (e.g., c-myc, AP-1, growth factors). Numerous reports have suggested that Ayurvedic plants and their components mediate their effects by modulating several of these recently identified therapeutic targets. However, Ayurvedic medicine requires rediscovery in light of our current knowledge of allopathic (modern) medicine. The focus of this review is to elucidate the Ayurvedic concept of cancer, including its classification, causes, pathogenesis and prevention; surgical removal of tumours; herbal remedies; dietary modifications; and spiritual treatments.

Animals↗

PubMed2005

Induction of hepatic cytochrome P450 isozymes, benzo(a)pyrene metabolism and DNA binding following exposure to polycyclic aromatic hydrocarbon residues generated during repeated fish fried oil in rats.

In the present study the effect of repeated fish fried oil (RFFO) and its extract (RFFE) on hepatic cytochrome P450 (CYP) isozymes, benzo(a)pyrene (BP) metabolism and DNA adduct formation was undertaken. HPLC analysis of RFFO showed the presence of several polycyclic aromatic hydrocarbons. CYP in microsomes from control and RFFO-treated animals showed a peak at 450 nm; however, a shift of 2 nm in the SORET region along with significant induction was observed in microsomes prepared from 3-methylcholanthrene (MC)- and RFFE-treated animals. Activities of hepatic ethoxyresorufin-O-deethylase, methoxyresorufin-O-deethylase, aryl hydrocarbon hydroxylase and erythromycin-N-demethylase were found to be significantly (P < 0.05) induced following exposure of RFFE, whereas none of these enzymes were altered in RFFO-treated group. Immunoblot analysis revealed that RFFE and MC were potent inducers of CYP1A1, 1A1/2 and 3A1 isozymes, where as RFFO showed no change in these protein levels. RT-PCR analysis showed induction of cDNA of CYP1A1 and CYP3A1 by RFFE treatment. Hepatic microsomes prepared from RFFE exposed animals enhanced BP metabolism with a concomitant increase in the relative proportion of BP 7,8-diol. Hepatic microsomes prepared from animals pretreated with RFFE and MC significantly enhanced the binding of [(3)H]-BP to calf thymus DNA. The overall results suggest that exposure to RFFE may induce hepatic CYP isozymes thereby producing enhanced reactive metabolites with a potential to bind with DNA that may result in cancer.

Animals↗

PubMed2005

In vitro cytotoxicity of polycyclic aromatic hydrocarbon residues arising through repeated fish fried oil in human hepatoma Hep G2 cell line.

Repeated frying of vegetarian and non-vegetarian foods in edible oil is a common practice round the globe. Our studies suggest that repeated fish fried oil (RFFO) generates polycyclic aromatic hydrocarbons (PAHs), which may lead to hazardous effect on human health. In order to understand the mechanism of toxicity of RFFO extracts containing a mixture of PAHs, the in vitro cytotoxicity assays in human hepatoma cell line, Hep G2 was undertaken. In addition to RFFO extract, benzo(a)pyrene (BP) and chrysene were used as prototype compounds for heavy and light PAHs, respectively. Doses of BP and chrysene were made in such a way, that it could represent the appropriate content of heavy and light PAHs found in the RFFO extract. Out of total content of PAHs (1240.4 microg/kg) in RFFO, major composition is of light PAHs (854.8 microg/kg) while heavy PAHs showed the concentration of 385.7 microg/kg. Treatment of cells with 1 microg/ml RFFO extract for 48 h showed significant induction in ethoxyresorufin-O-deethylase (EROD) activity. Exposure of cells to higher doses of RFFO extract (10-100 microg/ml) for 24, 48 and 72 h caused 3.5-5.2, 4.3-8.5 and 1.8-2.3-fold enhancement in EROD activity, respectively. Further, RFFO extract caused a dose dependent increase (2.1-3.5-fold) in aryl hydrocarbon hydroxylase (AHH) activity at 48 h. Induction of EROD and AHH activity in Hep G2 cells was found to be relatively more following BP or chrysene treatment as compared to RFFO extract. RFFO extract did not cause any significant effect on cell viability at 1 microg/ml and 10 microg/ml. However, at 100 microg/ml concentration RFFO extract significantly decreased the cell viability at 24, 48 and 72 h. Exposure of 10 microg/ml RFFO extract reduced the colony forming ability (CFA) of Hep G2 cells with maximum decrease of 33.5% at 72 h. However, exposure of cells to RFFO extract at highest concentration of assay (100 microg/ml) reduced CFA (35-52%) at 24, 48 and 72 h. RFFO extract (1-100 microg/ml) had no significant effect on growth inhibition of cell up to 48 h of exposure. However, exposure of RFFO extract at all doses showed significant growth inhibition (20-25%) at 72 h. In conclusion, the results suggest that RFFO extract has substantial cytotoxic potential through the metabolic activation process of PAHs generated per se.

Animals↗

PubMed2005

Dual control mechanism in a Belousov-Zhabotinskii (B-Z) oscillator with glucose and oxalic acid as a double substrate.

Oscillations in a Belousov-Zhabotinskii (B-Z) system having oxalic acid (OA) and glucose (G) as a mixed organic substrate, neither of which acts as a bromine scavenger, have been investigated. Studies have been performed for (i) varying the concentration of G while keeping the OA concentration fixed and (ii) varying OA but keeping G fixed in a batch reactor. In both cases upper and lower critical limits occur, between which oscillations are observed. Both single and double frequency oscillations have been observed in a wide range of concentrations of G as well as of OA. The induction period in most of the cases was <1 min. When G is fixed and OA is varied, the time pause between the sequential oscillations increases with an increase in OA. On the other hand when OA is fixed and G is varied, the time-pause decreases with an increase in G. The first type of oscillation is Br(-)-controlled, whereas the second is non-Br(-)-controlled. The order of addition of G and OA in the last has no influence on the induction period. It influences, however, the oscillatory characteristics. Br(2) evolution in the G + OA + Ce(4+) + BrO(3)(-) + H(2)SO(4) reaction system has been investigated spectrophotometrically. ESR and polymerization studies indicate the important role of free radicals in influencing the reaction mechanism. A tentative dual control mechanism has been suggested involving autocatalysis of HBrO(2) and BrO2*.

Journal Article↗

PubMed2002

Humoral immune response to an allogenic foetus in normal fertile women and recurrent aborters.

BACKGROUND: Several humoral immune factors are responsible for a successful pregnancy. There are a few studies, which demonstrate the role of antipaternal cytotoxic antibodies (APCA) and mixed lymphocyte reaction blocking factor (MLR-Bf) in the maintenance of pregnancy. However, these factors have not been studied in women with history of recurrent spontaneous abortion (RSA). We designed this study to review the role of APCA and MLR-Bf in normal pregnancy as well as in women with a history of RSA. METHODS: One hundred and five women with unexplained recurrent spontaneous abortion were included in the present study. These women were screened for all other known causes of recurrent abortion. We also included 60 normal fertile women, fifteen from each trimester and fifteen women during the post partum period (up to six months). RSA and controls (normal pregnancy) were matched for age, caste, and socio economic background and also for parity. APCA and MLR-Bf were evaluated in all the groups. All women with RSA who conceived during the study period were on follow up. RESULTS: We have analyzed the status of APCA and MLR-Bf in normal pregnancy (different gestational periods and during post partum), and in women with history of RSA. Our results show that APCA was significantly higher in controls as compared to RSA women. MLR-Bf was directed against the husbands' cells in normal pregnancy and was virtually absent in RSA women. CONCLUSION: Our results indicate that there is a significantly low titer of APCA and MLR-Bf in women with recurrent spontaneous abortion. This highlights the role of these factors in the maintenance of successful pregnancy.

Journal Article↗