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W Troll

Publications and source records attributed to W Troll.

At least 37 records · Page 2Linked to original sources
PubMed1986

Copper ions and hydrogen peroxide form hypochlorite from NaCl thereby mimicking myeloperoxidase.

Sea urchins have elaborated multiple defenses to assure monospermic fertilization. In this work, we have concentrated on a study of the mechanism(s) by which hydrogen peroxide (H2O2) prevents polyspermy in Arbacia punctulata. We found that it is not H2O2 but probably hypochlorous acid/hypochlorite (HOCl/OCl-) derived from H2O2 that is toxic to the supernumerary sperm. The spermicidal activity of H2O2 is potentiated by at least one order of magnitude by cupric ions (Cu2+). This increased toxicity is not due to the formation of hydroxyl radicals (.OH) because .OH scavengers did not counteract the activity of Cu2+. Moreover, substitution of Cu2+ by ferrous ions (Fe2+), which are known to cause formation of .OH from H2O2, had no effect on fertilization even at 10(2)-10(3) times higher concentrations. In contrast, 3-amino-1,2,4-triazole (AT), and HOCl/OCl- scavenger, totally reversed the toxic effects of Cu2+. Furthermore, we found that HOCl/OCl- is generated in solutions of H2O2 and Cu2+ in the presence of 0.5 M NaCl and that its accumulation is abolished by AT. Thus it is possible that the antifertility properties of copper are due to its ability to mediate formation of HOCl/OCl-. HOCl/OCl- generated by Cu2+ from H2O2 and Cl-, a low concentration of exogenously added HOCl/OCl-, or increased concentrations of H2O2 has similar inhibitory effects on the fertilization process in sea urchins. Therefore, we suggest that polyspermy is prevented by the action of a myeloperoxidase that affects the formation of HOCl/OCl- from the Cl- present in sea water through reaction with H2O2 generated by the newly fertilized egg.

Amitrole↗
PubMed1986

Some protease inhibitors are also inhibitors of poly(ADP-ribose) polymerase.

The low-molecular-weight peptide protease inhibitors, tosyl-lysine-chloromethyl ketone, antipain and leupeptin, inhibited poly(ADP-ribose) [poly(ADP-Rib)] polymerase in permeable cells. The concentrations required for 50% inhibition were 3.6, 5 and 29 mM, respectively. Two peptides without protease inhibitor activity, fibrinopeptide A and phenylalanine-leucine-(glutamine)2-leucine, also inhibited poly (ADP-Rib) synthesis; doses required for 50% inhibition were 0.37 and 11.2 mM, respectively. These concentrations lie within a range bracketed by the 50% inhibition concentrations of the strong and weak poly(ADP-Rib) synthesis inhibitors, 3-amino-benzamide (0.15 mM) and caffeine (greater than 100 mM), respectively. N-Ethylmaleimide also inhibited poly(ADP-Rib) synthesis, at a 50% inhibitory dose of 0.3 mM, in the absence of exogenous thiol reagents. High-molecular-weight protease inhibitors, such as soybean (including Bowman-Birk reagent) and lima bean trypsin inhibitors and human alpha 1-protease inhibitor, had no effect on poly(ADP-Rib) synthesis up to 2 mg/ml. Interference with transformation and other cellular effects that have been reported in carcinogen-damaged cells treated with low-molecular-weight peptide protease inhibitors may therefore involve common mechanisms with poly(ADP-Rib) inhibitors. Similar effects of high-molecular-weight protease inhibitors presumably involve different mechanisms.

Antipain↗
PubMed1986

Radiation-like modification of bases in DNA exposed to tumor promoter-activated polymorphonuclear leukocytes.

Oxygen species generated by human polymorphonuclear leukocytes (PMNs) activated by 12-O-tetradecanoylphorbol-13-acetate (TPA) caused the formation of 5-hydroxymethyl-2'-deoxyuridine (HMdUrd), and (+) and (-) diastereoisomers of cis-thymidine glycol (dTG) in DNA that was exposed to them. There were 9 HMdUrds and 31 dTGs formed per 1 X 10(6) thymidine residues. When Fe(II)/ethylenediaminetetraacetic acid was added to TPA-activated PMNs at 0, 10, 15, and 20 min after TPA, HMdUrd formation increased 5-, 13-, 30-, and 35-fold. Although dTG was initially formed in larger amounts than HMdUrd, it eventually decreased but was still 5-, 6-, 5.5-, and 3-5-fold, respectively, higher than in the absence of iron. From 65 to 1800 times more HMdUrd was formed in DNA when autologous plasma was present during incubation of DNA with TPA-activated PMNs than in its absence. The levels of dTG also varied from about the same as HMdUrd to the nondetectable. Reconstituted human serum transferrin used instead of plasma or Fe(II) also supported the formation of HMdUrd and dTG. When DNA was treated with Fe(II)-reduced H2O2 in the absence of PMNs and TPA, both derivatives were formed. However, the same treatment of marker dTG of dTG-containing polydeoxyadenylic-thymidylic acid caused the decomposition of dTG. Thus, the reduction of hydrogen peroxide by Fe(II) complexed to either ethylenediaminetetraacetic acid or amino acids amy be responsible for the formation of HMdUrd and dTG and for subsequent decomposition of dTG in DNA exposed to the TPA-activated PMNs.

DNA↗
PubMed1985

Expression of long terminal repeat (LTR) sequences in carcinogen-induced murine skin carcinomas.

RNA sequences homologous to the Long Terminal Repeat (LTR) sequence of Moloney Murine Leukemia Virus proviral DNA are expressed in murine squamous cell carcinomas of the skin induced by chemical carcinogens. These transcripts range in size from 8.2 to less than 2.4 kb but their size profile varies between individual tumors. These RNAs are not detected in the poly A+ RNA fraction obtained from the epidermis of control mice or carcinogen induced skin papillomas. The poly A+ RNAs from the livers and spleens of some of the mice with skin carcinomas also revealed LTR related sequences, whereas these RNAs were not detected in the livers and spleens of control mice or of carcinogen-treated mice that did not develop carcinomas. Thus, chemical carcinogenesis in mouse skin is associated with constitutive expression of endogenous retrovirus related sequences in the carcinomas as well as in certain apparently normal host tissues.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1985

Nanomolar concentrations of Bowman-Birk soybean protease inhibitor suppress x-ray-induced transformation in vitro.

Experiments reported here indicate a crude soybean extract, if defatted with acetone, effectively blocks cell transformation in vitro. An active component of this crude extract is the Bowman-Birk trypsin and chymotrypsin inhibitor. The chymotrypsin-inhibitory region of the Bowman-Birk inhibitor is responsible for suppressing in vitro transformation. Another low molecular weight soybean trypsin inhibitor does not significantly suppress transformation. The Bowman-Birk inhibitor (i) has an irreversible effect on the transformation process, (ii) can suppress radiation-induced transformation even when added to cultures many days after the carcinogen exposure, and (iii) is effective in its ability to suppress transformation when present in the medium at a concentration as low as 0.125 nM.

Acetone↗
PubMed1984

Role of free radicals in the initiation and promotion of radiation transformation in vitro.

We have studied the effects of superoxide dismutase (SOD), catalase, Cu(II) (3,5-diisopropylsalicylate)2 (CuDIPS) and other copper compounds on radiation transformation in vitro using C3H 10T1/2 cells. When present only during irradiation, high concentrations of SOD in the medium enhanced transformation, while catalase, inactivated SOD (autoclaved), CuDIPS, cupric chloride and cuprous chloride inhibited the initiation phase of radiation transformation. SOD, catalase and CuDIPS did not affect the expression phase of radiation transformation. Suppression of the TPA enhancement of transformation by catalase was a highly significant effect, while the suppression by SOD was not of statistical significance. Our results suggest that hydrogen peroxide (H2O2) may be important in the cellular damage leading to malignant transformation.

Animals↗
PubMed1983

Inhibition of superoxide production in human neutrophils by purified soybean polypeptides. Re-evaluation of the involvement of proteases.

Inhibition of neutrophil superoxide production has been previously reported for reagents and polypeptides which also inhibit serine proteases. There are disagreements between the results of different laboratories including our own, which have attempted to use the Kunitz soybean trypsin inhibitor to block neutrophil superoxide production. Having confirmed that crude extracts of soybean do contain inhibitory factors which affect neutrophil superoxide production, we have resolved polypeptides in an ethanolic extract of soybean flour by anion and cation exchange chromatography and preparative polyacrylamide gel electrophoresis. Fractions have been assayed for protease inhibitory activity and inhibition of neutrophil superoxide production. We have found an inverse relation between these two inhibitory activities during the purification process. Two of three isolated polypeptides are potent inhibitors of neutrophil superoxide production (50% inhibition at 10(-7) M) but retain only weak anti-trypsin activity. A third polypeptide is a potent inhibitor of trypsin but is completely lacking superoxide inhibitory activity. None of the isolated polypeptides inhibit chymotrypsin. The implications of these findings for the hypothetical association between neutrophil production of superoxide and cellular proteases are discussed.

Electrophoresis, Polyacrylamide Gel↗
PubMed1983

Murine lymphocyte cell surface proteolytic activity is strain related.

A/J and C57 Br/cdj mice bear the H-2 haplotypes which are generally associated with high responses to bovine gamma globulin and type 3 pneumococcal polysaccharide. A/J has been reported, however, to produce higher levels of antibody than C57 Br/cdj mice against the antigens. The two strains of mice were used as model systems in this study to determine whether the level of lymphoid cell surface proteolytic activity is also genetically controlled. The results of this study illustrate that the level of lymphoid cell surface proteolytic activity is strain related. Since A/J lymphocytes were found to have a significantly higher rate of proteolysis than C57 Br/cdj lymphocytes, a correlation between lymphoid cell surface caseinolytic activity and immune responsiveness is suggested.

Animals↗
PubMed1983

Reevaluation of the presence of the major antigen Ca++ complex in Bernard-Soulier syndrome platelets. Elastase degradation of the complex in Bernard-Soulier syndrome platelet preparations.

Crossed immunoelectrophoresis of human platelet membranes reveals a major antigen GPIIb-GPIIIa-Ca++ complex which is absent to decreased in Glanzmann's thrombasthenia. In our initial report, we also noted its absence in two patients with Bernard-Soulier syndrome associated with the presence of two neoantigens, and postulated that this may be due to endogenous proteolysis. It now appears that this observation is due to exogenous proteolysis from granulocyte-elastase contamination in the platelet membrane preparation. Purified elastase is capable of degrading the major antigen-Ca++ complex, particularly in its dissociated form, to the pattern initially reported for Bernard-Soulier platelets. Of particular interest is the observation that the elastase-degraded dissociated complex can be reassociated in the presence of Ca++. This indicates that the elastase-sensitive antigenic determinants recognized by the rabbit anti-human platelet membrane antibody are not required for the bonds which reassociate the complex.

Blood Coagulation Disorders↗
PubMed1983

Protease inhibitors: possible anticarcinogens in edible seeds.

Protease inhibitors, which are common constituents of seeds (rice, beans, and maize) have been shown to inhibit breast, colon, and skin cancers in animal experiments. Epidemiological studies have shown that diets rich in these components (ie seed proteins) decrease the occurrence of prostatic, breast, and colon cancers in man. We have demonstrated that a typical purified inhibitor from soybeans, the Bowman-Birk inhibitor, is excreted in the feces complexed with digestive proteases from the duodenum. This results in greater excretion of proteins, thus effectively decreasing the amount of protein available for digestion. Excess protein in the diet in addition to fats may contribute to increased cancer. Thus, the addition of seeds (chick-peas, beans, etc) to the diet rather than removal of meat or fat offers the opportunity of preventing these cancers, because the protease inhibitors present would effectively remove a portion of the proteins being consumed. Protease inhibitors also act directly as antioxidants in relation to tumor promoters and ionizing radiation. To what extent this contributes to the nutritional effects of eating seeds containing protease inhibitors remains to be determined.

Animals↗
PubMed1983

Modifiers of free radicals inhibit in vitro the oncogenic actions of x-rays, bleomycin, and the tumor promoter 12-O-tetradecanoylphorbol 13-acetate.

Using short-term cultures of hamster embryo cells, we have examined the effects of the free-radical scavenger superoxide dismutase (superoxide:superoxide oxidoreductase, EC 1.15.1.1) and the enzyme catalase (hydrogen-peroxide:hydrogenperoxide oxidoreductase, EC 1.11.1.6) on x-ray- and bleomycin-induced transformation and on the enhancement of radiogenic transformation by the tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA). We find that superoxide dismutase inhibits (i) transformation induced by x-ray and bleomycin and (ii) promotional action of TPA in vitro. The results suggest that the oncogenic action of x-rays and bleomycin and the enhancement of oncogenic transformation by TPA are mediated in part by free radicals. The findings also suggest that superoxide dismutase can serve as an inhibitor of oncogenesis and that its actions, as seen in this in vitro system, are most predominantly on inhibiting late events in the progression of cellular transformation--those associated with promotion.

Animals↗
PubMed1983

Effect of inhibitors of trypsin-like proteolytic enzymes Bacillus cereus T spore germination.

The germination of Bacillus cereus T spore suspensions is partially prevented by several inhibitors of trypsin-like enzymes. Leupeptin, antipain, and tosyl-lysine-chloromethyl ketone are effective inhibitors, whereas chymostatin, elastatinal, and pepstatin are inactive. A synthetic substrate of trypsin, tosyl-arginine-methyl ester, also inhibits germination. Its inhibitory effect decreases as a function of incubation time in the presence of spores and is abolished by previous hydrolysis with trypsin. Germinating, but not dormant, spore suspensions hydrolyze tosyl-arginine-methyl ester; its hydrolysis is insensitive to chloramphenicol, sulfhydryl reagents, and EDTA. A crude extract of germinated B. cereus spores contains a trypsin-like enzyme whose activity, as measured by hydrolysis of benzoyl-arginine p-nitroanilide, is sensitive to germination-inhibitory compounds such as leupeptin, tosyl-arginine-methyl ester, and tosyl-lysine-chloromethyl ketone. Spore suspensions exposed to the above inhibitors under germination conditions lose only part of their heat resistance and some 10 to 30% of their dipicolinic acid content. Part of the germinating spore population becomes "phase grey" under phase optics. Based on a study of the inhibition of germination by protease inhibitors and the activity of a protease in germination spores and spore extracts, it is suggested that the activity of a trypsin-like enzyme may be involved in the mechanism of the breaking of dormancy in spores of B. cereus T.

Antipain↗
PubMed1983

Free oxygen radicals: necessary contributors to tumor promotion and cocarcinogenesis.

The two stage model of carcinogenesis postulates that agents which lead to tumor formation are either initiating or promoting agents. Initiating agents induce chemical modifications of DNA. Promoting agents have been assumed to induce cell proliferation and thereby establish a milieu in which the initiated cells can express their irreversibly altered genotype. Promoters can induce both the migration of leukocytes to the skin and stimulate their respiratory activity leading to the formation of active O2 species which cause lipid peroxidation and thereby change membrane properties. However, it was recently found that the O2 species emanating from promoter-activated leukocytes also cause formation of strand breaks both in the DNA of the leukocytes themselves and in the DNA of cells cocultivated with such leukocytes. The contribution of activated O2 species to tumor promotion has been confirmed by the finding that antioxidants and copper containing superoxide dismutase (SOD)-mimetic compounds, protease inhibitors and retinoids (which block free radical formation) inhibit promotion. Furthermore, cocarcinogens such as gossypol and pyrogallol may also cause conversion of O2 to the superoxide anion radical. The heritable change caused by initiators may render the cell more vulnerable to promoter-mediated DNA damage. Experimental investigation of these hypotheses may elucidate our understanding of the contribution of promoter-mediated DNA damage in tumorigenesis and identify preventive agents for carcinogenesis.

Animals↗
PubMed1983

Bowman-Birk soybean protease inhibitor as an anticarcinogen.

Vegetarian populations show a decreased occurrence of breast, colon, and prostatic cancers. Epidemiological studies have identified seeds (maize, corn, and beans) as protective agents in these cancers. We have selected to study one abundant component of all seeds, protease inhibitors. Synthetic and natural protease inhibitors have been shown to inhibit tumor promotion in vivo and in vitro. In the present study, we report that a typical, natural protease inhibitor, the Bowman-Birk inhibitor isolated from soybeans, survives inactivation by stomach digestion in rodents and appears to be fully active as a protease inhibitor in the small intestine, where it complexes with the proteases occurring there, i.e., trypsin and chymotrypsin. A large part of the inhibitor is excreted as protease:protease inhibitor complexed in the feces. We also report the specific inhibition of transformation caused by ionizing radiation by this protease inhibitor. The mechanism of anticarcinogenesis of ingested protease inhibitors may involve the indirect effect of partially blocking protein absorption. High-protein and high-fat diets are known to increase cancer occurrence. Protease inhibitors reaching specific sites also have anticarcinogenic activities, as demonstrated by the radioprotective effect of a protease inhibitor in vitro. The relative importance of the indirect and direct action of protease inhibitors remains to be established.

Animals↗