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Biomedical subjects

M Das

Publications and source records attributed to M Das.

At least 217 records · Page 12Linked to original sources
PubMed1986

Benzo(a)pyrene diol epoxide-I-DNA adduct formation in the epidermis and lung of SENCAR mice following topical application of crude coal tar.

The levels of benzo[a]pyrene diol epoxide-I-deoxyguanosine (BPDE-I-dG) adduct formation in epidermis and lung of SENCAR mice following the topical application of benzo[a]pyrene (BP) alone, crude coal tar (CCT) alone, and the two combined were determined in an enzyme linked immunosorbent (ELISA) assay using monoclonal antibodies. Topical application of two doses of BP (20 micrograms) at 72-h intervals, with sacrifice 24 h later resulted in the formation of 197 fmol and 205 fmol BPDE-I-dG adducts per mg DNA in epidermis and lung, respectively. Topical application of 0.5 ml CCT alone resulted in the formation of 278 fmol and 410 fmol BPDE-I-dG adducts per mg DNA in epidermis and lung, respectively. Simultaneous topical application of 20 micrograms BP and CCT (0.1-0.5 ml) resulted in substantially lower BPDE-I-dG adducts in the epidermis as well as in the lung. Our results suggest that CCT may contain inhibitors of carcinogen-DNA adduct formation and that topical application of CCT produces greater effects on DNA-adduct formation in lung than in epidermis. Thus the cancer-causing potency of the polycyclic aromatic hydrocarbons (PAHs) in CCT may be reduced by other anticarcinogenic constituents present in CCT and systemic absorption of carcinogenic PAHs in CCT applied to skin might have tumorigenic effects in other tissues.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1986

Experimental infection of rhesus monkeys with Schistosoma incognitum and Orientobilharzia dattai.

Rhesus monkeys inoculated subcutaneously with 800-2500 cercariae of Schistosoma incognitum did not yield adult worms. Percutaneous exposures, however, resulted in the presence of adult flukes 21-35 days post infection. The infection became very mild (0-4 flukes) 45-100 days after exposure and no eggs were excreted. Monkeys percutaneously exposed to cercariae of Orientobilharzia dattai and treated with prednisolone yielded a very small number of flukes 22 days after exposure while non treated controls yielded no adult worms.

Animals↗
PubMed1986

Interaction of acrylamide with bovine serum albumin.

The binding of acrylamide (ACR) with purified bovine serum albumin (BSA) was studied. Binding of ACR with BSA was characterized by equilibrium dialysis, fluorescence studies, and ultraviolet spectroscopy. ACR was quantitated by high-pressure liquid chromatography. Equilibrium dialysis studies on the binding of ACR with BSA showed that more than 25% of the added ligand was bound to the protein at equilibrium. ACR produced a concentration-dependent decrease in uv absorbance of BSA, indicating reactivity of ACR with BSA. Fluorescence quenching studies of ACR binding showed a concentration-dependent quenching of the fluorescence of BSA. ACR also caused a concentration-dependent decrease in the fluorescence of sulfhydryl (-SH) groups present on BSA, implicating a role of protein -SH groups in the binding of ACR. Prior blocking of -SH groups by N-ethylmaleimide resulted in 16% inhibition of the initial binding of ACR, suggesting involvement of -SH groups. Our results demonstrate that ACR binds to BSA through both aromatic amino acids and -SH groups and that such binding may play an important role in pharmacodynamics and toxicity of ACR.

Acrylamide↗
PubMed1986

Perinuclear location and recycling of epidermal growth factor receptor kinase: immunofluorescent visualization using antibodies directed to kinase and extracellular domains.

This paper describes studies on the migratory behavior of epidermal growth factor (EGF) receptor kinase using antibodies that are specific for either the kinase domain or the extracellular domain of the receptor. Antiserum was raised to a 42,000-D subfragment of EGF receptor, which was shown earlier to carry the kinase catalytic site but not the EGF-binding site. Another antiserum was raised to the pure intact 170,000-D EGF receptor. The specificities of these antibodies were established by immunoprecipitation and immunoblotting experiments. The domain specificity was examined by indirect immunofluorescent staining of fixed cells. The anti-42-kD peptide antibody could bind specifically to EGF receptors of both human and murine origin and was found to be directed to the cytoplasmic part of the molecule. It did not bind to EGF receptor-negative cells, which contained other types of tyrosine kinases. The antibodies raised against the intact receptor recognized only EGF receptor-specific epitopes and were directed to the extracellular part of the molecule. The anti-receptor antibodies described above were used to visualize the cyclic locomotory behavior of EGF receptor kinase under various conditions of EGF stimulation and withdrawal. The receptor was examined in fixed and permeabilized cells by indirect immunofluorescent staining. The results demonstrate the following: (a) the receptor kinase domain migrates to the perinuclear region upon challenge with EGF; (b) both extracellular and cytoplasmic domains of the receptor are involved in migration as a unit; (c) withdrawal of EGF results in rapid recycling of the perinuclear receptors to the plasma membrane; (d) this return to the cell surface is inhibited by methylamine, chloroquine, and monensin; and (e) neither the internal migration nor the recycling process is blocked by inhibitors of protein biosynthesis.

Animals↗
PubMed1986

Clotrimazole, an inhibitor of benzo[a]pyrene metabolism and its subsequent glucuronidation, sulfation, and macromolecular binding in BALB/c mouse cultured keratinocytes.

The effect of the antifungal imidazole compound, clotrimazole, on the metabolism of benzo[a]pyrene (BP) was studied in cultured keratinocytes prepared from BALB/c mouse epidermis. Varying concentrations of clotrimazole added to the cultured keratinocytes resulted in a dose-dependent inhibition of the activities of the microsomal cytochrome P-450-dependent monooxygenases aryl hydrocarbon hydroxylase and 7-ethoxycoumarin O-deethylase. The major organic solvent-soluble metabolites of BP identified in the cultured cells were trans-7,8-dihydro-7,8-dihydroxybenzo[a]pyrene (BP-7,8-diol), 9-hydroxybenzo[a]pyrene (9-OH-BP), and 3-hydroxybenzo[a]pyrene (3-OH-BP), although small amounts of trans-4,5-dihydro-4,5-dihydroxybenzo[a]pyrene, BP-quinones, and trans-9,10-dihydroxybenzo[a]pyrene were also present. The major organic solvent-extractable metabolites of BP found in the extracellular culture medium were primarily the diols with smaller quantities of phenols and quinones. The major water-soluble metabolites of BP present both intracellularly and extracellularly were glucuronide conjugates of 3-OH-BP, 9-OH-BP, and benzo[a]pyrene-3,6-dione and to a lesser extent sulfate conjugates (primarily of the BP-7,8-diol). Clotrimazole inhibited the generation of organic solvent-soluble and water-soluble conjugates in a dose-dependent manner. The in vitro metabolism of BP by microsomes prepared from control and benz[a]anthracene (BA)-induced cultured keratinocytes was also inhibited by clotrimazole with greater inhibitory effect on BA-induced keratinocytes especially with respect to the formation of diols and quinones. The enzyme-mediated covalent binding of BP to mouse keratinocyte DNA and protein was also substantially diminished by clotrimazole in a dose-dependent fashion. These results indicate that clotrimazole, a widely used drug for the management of a variety of superficial dermatophyte infections of the skin, is a potent inhibitor of cytochrome P-450-dependent transformation of polycyclic aromatic hydrocarbons in cultured murine keratinocytes. This system offers a convenient approach for studies as inhibitors of carcinogen metabolism in the epidermis.

Animals↗
PubMed1986

Additive effects of ultraviolet B and crude coal tar on cutaneous carcinogen metabolism: possible relevance to the tumorigenicity of the Goeckerman regimen.

The effect of cutaneous exposure to ultraviolet B (UVB) radiation alone, to crude coal tar (CCT) alone, and to the combination of UVB and CCT on the inducibility of the microsomal cytochrome P-450-dependent carcinogen-metabolizing enzyme aryl hydrocarbon hydroxylase (AHH) and other monooxygenases such as 7-ethoxyresorufin O-deethylase (ERD) and 7-ethoxycoumarin O-deethylase (ECD) activities in the skin of neonatal rats was studied. Exposure of the animals to UVB (400-1600 mJ/cm2) alone resulted in a dose-dependent increase in cutaneous enzyme activities. At a UVB dose of 1200 mJ/cm2 increases in AHH, ECD, and ERD were 194%, 115%, and 244%, respectively. A single topical application of CCT (10 ml/kg) 24 h before sacrifice resulted in significant induction of AHH (350%), ECD (921%), and ERD (796%) activities. Treatment of animals with the same dose of CCT followed by UVB exposure resulted in additive and/or synergistic effects on AHH (858%), ECD (1229%), and ERD (1166%) activities in the skin. In contrast, exposure of animals to UVB prior to CCT application had effects no different from those of CCT alone. Epoxide hydrolase and glutathione S-transferase activities in skin from all experimental groups were not different from those of controls. High-pressure liquid chromatographic analysis of the metabolism of benzo[a]pyrene (BP) by cutaneous microsomes prepared from animals treated with UVB alone, CCT alone, and the combination of UVB and CCT revealed increased formation of all the metabolites in each experimental group. The largest increase in metabolite formation occurred in animals receiving CCT followed by UVB exposure. The inducibility of trans-7,8-diol formation by UVB alone and CCT alone was 203% and 435%, respectively, whereas with CCT followed by UVB it was 1065%. The differential responses in AHH activity were found to parallel the capacity of skin microsomal enzymes to enhance the binding of [3H]-BP to DNA. These studies indicate that the sequence of exposure to the components of the Goeckerman regimen in rodents greatly influences metabolic activity in skin. When applied in the same sequence employed in the Goeckerman regimen (CCT followed by UVB exposure) the additive effect upon catalytic activity essential for cancer initiation suggests a possible mechanism for the enhancement of human skin cancer in individuals exposed to this therapeutic regimen.

Animals↗
PubMed1986

Pharmacological modification of epidermal detoxification systems.

The skin is a major interface between the body and the environment and possesses membrane-bound cytochrome P-450-dependent enzyme activity that is capable of both detoxifying endogenous and exogenous substrates and enhancing the toxic effects of selected substances. The use of chemical inhibitors of enzyme activation by P-450-dependent enzymes represents one possible approach to controlling toxic responses in target tissue such as the skin. Our data indicate that certain polyphenols and imidazoles are potent inhibitors of epidermal carcinogen metabolism, and of the DNA binding and the carcinogenicity of PAHs such as BP. The use of such agents may offer a novel approach to the prevention of environmentally induced cancer.

Animals↗
PubMed1986

Inhibition of 3-methylcholanthrene-induced skin tumorigenicity in BALB/c mice by chronic oral feeding of trace amounts of ellagic acid in drinking water.

Chronic p.o. feeding of small amounts of ellagic acid, a naturally occurring dietary plant phenol, to BALB/c mice in drinking water afforded significant protection against skin tumorigenesis induced by 3-methylcholanthrene, a polycyclic aromatic hydrocarbon carcinogen. A significant increase in the latent period for the development of skin tumors by 3-methylcholanthrene was observed in the ellagic acid-fed group of mice (9 wk on test) as compared to the control group of animals (6 wk on test). The observed protection against tumor induction in the ellagic acid-fed group of animals may be due to the inhibition of the metabolic activation of the polycyclic aromatic hydrocarbon since epidermal aryl hydrocarbon hydroxylase activity was found to be significantly inhibited. Our results suggest that dietary supplementation with small amounts of ellagic acid may prove useful in reducing the risk of skin carcinogenesis induced by environmental chemicals.

7-Alkoxycoumarin O-Dealkylase↗
PubMed1985

Differential activities of rat and human lung glutathione S-transferase isoenzymes towards benzo(a)pyrene epoxides.

The isoenzymes of human and rat lung glutathione S-transferase (GST) differ among themselves in their activities towards the epoxides of benzo(a)pyrene (BP). The Ya' and Yc-type subunits of rat lung GST exhibit maximum activities towards BP-4,5-oxide and BP-7,8-oxide suggesting that these two subunits are preferentially involved in the detoxification of highly reactive epoxides and diol-epoxides of polycyclic aromatic hydrocarbons (PAH). The studies with human lung GST isoenzymes indicate that BP-4,5-oxide, and BP-7,8-oxide are preferred substrates for the cationic (pI 8.3) form of the enzyme. Identification of compounds which can selectively induce these isoenzymes of GST could prove useful as inhibitors of PAH induced neoplasia.

Animals↗
PubMed1985

The National Football Head and Neck Injury Registry. 14-year report on cervical quadriplegia, 1971 through 1984.

Data on cervical spine injuries resulting from participation in football have been compiled by a national registry. Analysis of epidemiologic data and cinematographic documentation clearly demonstrated that the majority of cervical fractures and dislocations were due to axial loading. On the basis of this observation, rule changes banning both deliberate "spearing" and the use of the top of the helmet as the initial point of contact in making a tackle were implemented at the high school and college level. Subsequently, a marked decrease in cervical spine injury rates has occurred. The occurrence of permanent cervical quadriplegia decreased from 34 in 1976 to five in the 1984 season. It is suggested that axial loading of the cervical spine is also responsible for the catastrophic injuries in diving, rugby, ice hockey, and gymnastics. Implementation of appropriate changes in playing techniques and/or equipment modifications could possibly reduce the incidence of cervical spine injuries in these activities.

Athletic Injuries↗
PubMed1985

Intrapeptide autophosphorylation of the epidermal growth factor receptor: regulation of kinase catalytic function by receptor dimerization.

The epidermal growth factor (EGF) receptor is a transmembrane polypeptide of 170 000 daltons (Da) with a cytoplasmically facing protein kinase domain. The regulation of the tyrosine kinase activity of the EGF receptor by added EGF and by receptor association state was studied in an in vitro system. The rate of autophosphorylation of the solubilized and purified EGF receptor was found to be independent of receptor concentration. To determine whether the zero-order kinetics observed point to intrapeptide phosphorylation, we measured the sedimentation characteristics of the undenatured solubilized receptor. The receptor was found to exist in two association-dissociation states-a monomeric 7.7S form and a dimeric 12S form. The 7.7S form is an active tyrosine kinase; it has high basal activity, and the activity is not further stimulated by EGF; it appears to be an EGF-independent form of the receptor kinase. The 12S form is devoid of catalytic activity, but in the presence of EGF it dissociates into the active monomeric form. Freshly purified receptor preparations contain mainly the monomeric receptor, have high basal kinase activity, and show low EGF stimulatability (less than 1.3-fold). Aging of the receptor results in progressive dimerization and decay of EGF-independent kinase activity (and increase in EGF stimulatability). All of these processes are reversed in the presence of EGF or dithiothreitol.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Topically applied nitropyrenes are potent inducers of cutaneous and hepatic monooxygenases.

The inducibility of skin and liver microsomal cytochrome P-450 dependent aryl hydrocarbon hydroxylase and other monooxygenases by a mixture of nitropyrenes was assessed and compared with the parent non-nitrated compound, pyrene. A single topical application of nitropyrenes to neonatal rats resulted in highly significant induction of aryl hydrocarbon hydroxylase, ethoxycoumarin O-de-ethylase, and ethoxyresorufin O-de-ethylase activities in skin and liver after 24 hours. Inducibility of the skin and liver enzymes was 3.9-5.7 fold and 1.8-10.3 fold respectively. On the other hand, aminopyrine N-demethylase, benzphetamine N-demethylase and epoxide hydrolase activities in the liver were unaffected by topically applied nitropyrenes. Furthermore, treatment with nitropyrenes produced a 1 nm shift to the blue region in the wavelength maximum of hepatic microsomal cytochrome P-450. Topically applied pyrene produced only marginal or no effects on cutaneous and hepatic enzyme activities. Our results suggest that nitration of pyrene, a relatively ineffective enzyme inducer, produces nitropyrenes which are potent inducers of hepatic and cutaneous monooxygenases and they resemble 3-methylcholanthrene in this inducing effect.

7-Alkoxycoumarin O-Dealkylase↗