DNA base and deoxyribose modification by the carbon-centered radical generated from 4-(hydroxymethyl)benzenediazonium salt, a carcinogen in mushroom.

Modification of the base and the sugar moieties of DNA with 4-(hydroxymethyl)benzenediazonium salt (HMBD), a carcinogen in the mushroom Agaricus bisporus, was investigated. When deoxyribonucleosides dGuo, dAdo, dThd, and dCyd were incubated with HMBD at pH 7.4 and 37 degrees C, the levels of all the nucleosides were decreased.

Induction of DNA recombination by activated 3-amino-1-methyl-5H-pyrido[4,3-b]indole.

To investigate the genotoxic properties of a food-derived carcinogen, 3-amino-1-methyl-5H-pyrido-[4,3-b]indole (Trp-P-2), we have tested whether Trp-P-2 and its metabolically transformed products can induce DNA recombinations. Trp-P-2 is a strong mutagen and its activated form, the N-hydroxylated derivative, Trp-P-2(NHOH), is known to form DNA adducts and cause DNA chain cleavage. Using a system in which phage lambda undergoes recombination inside host Escherichia coli, we have found that Trp-P-2(NHOH), but not Trp-P-2 itself, can induce recombination.

DNA strand breaking by the carbon-centered radical generated from 4-(hydroxymethyl) benzenediazonium salt, a carcinogen in mushroom Agaricus bisporus.

4-(Hydroxymethyl)benzenediazonium salt (HMBD), a carcinogen in mushroom Agaricus bisporus, was found to generate a carbon-centered radical, 4-(hydroxymethyl)phenyl radical, during incubation at pH 7.4 and 37 degrees C, when estimated by Electron Spin Resonance (ESR) spin-trapping technique using 5,5-dimethyl-1-pyrroline N-oxide (DMPO), N-tert-butylphenyl-alpha-nitrone (PBN) and 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS). Formation of a substantial amount of benzyl alcohol during incubation of HMBD in the presence of a hydrogen donor, ethanol, supported the generation of the carbon-centered radical.

Enhancement of nitrogen dioxide-induced lipid peroxidation and DNA strand breaking by cysteine and glutathione.

Nitrogen dioxide less than 100 ppm in air induced lipid peroxidation of liposome composed of 1-palmitoyl-2-arachidonylphosphatidylcholine as assessed by thiobarbituric acid reactivity. The nitrogen dioxide-induced lipid peroxidation was enhanced by cysteine, glutathione and bovine serum albumin. While the activity of nitrogen dioxide in air to induce single strand breaks of supercoiled plasmid DNA was low, the breaking was remarkably enhanced by cysteine, glutathione and bovine serum albumin.

Appearance of ESR signals by the reaction of 3,5-dibromo-4-nitrosobenzenesulfonate (DBNBS) and non-radical biological components.

The reaction of 3,5-dibromo-4-nitrosobenzenesulfonate (DBNBS) with non-radical biological components produced spin adducts with ESR signals. The reactions of DBNBS with Trp, Gly-Trp, Trp-Gly, Pro, Cys and glutathione at pH 7.5 and room temperature for more than 1 hour gave the nitroxyl free radicals with ESR signals, whereas the reactions with other amino acids and bovine serum albumin did not.

Mechanisms of the DNA breaking activity of mutagenic 5-diazouracil.

5-Diazouracil in monohydrated form showed mutagenicity and cytotoxicity on Salmonella typhimurium TA98 and TA100 strains without metabolic activation, and induced mouse micronucleated peripheral reticulocytes. Incubation of a plasmid supercoiled DNA with the compound caused DNA single-strand breaking: the supercoiled form was transformed into an open circular relaxed form and then into a linear form. The breaking was similarly caused in the absence of molecular oxygen.

Possible occurrence of new mutagens with the DNA breaking activity in coffee.

Brewed and instant coffee emitted strong chemiluminescence due to singlet oxygen and excited carbonyls, which may be originated by the Maillard reaction of sugars and amino acids but not by the reaction of polyphenolics. Instant coffee cleaved DNA giving single-strand breaks only after it was purified by high performance liquid chromatography (HPLC) or by gel filtration. Retention times of component(s) with strong DNA breaking activity in HPLC were different from those of chemiluminescence emitters, although they were coeluted on a gel filtration.

Generation of DNA-breaking activity in the Maillard reaction of glucose-amino acid mixtures in a solid system.

Maillard products of 1:1 solid mixtures of glucose-amino acid heated at 200 degrees C for 5 min induced single-strand breaks of DNA after incubation at 37 degrees C and pH 7.4 overnight. The products of Gly, His, Trp, Tyr, Phe and CySH transformed a plasmid supercoiled DNA (form I) into an open circular relaxed form (form II) and a linear form (form III).

DNA breaking activity of the carbon-centered radical generated from 2,2'-azobis(2-amidinopropane) hydrochloride (AAPH).

When supercoiled plasmid DNA was incubated with 2,2'-azobis (2-amidinopropane)hydrochloride (AAPH) at pH 7.4 in the presence and absence of oxygen, the DNA single strands were effectively cleaved. The breaking in the presence of oxygen was not inhibited by superoxide dismutase and catalase, but inhibited by mannitol, ethanol, butyl hydroxyanisole, thiol compounds, tertiary amines and spin trapping agents N-tert-butyl-alpha-phenylnitrone (PBN) and 5,5-dimethyl-1-pyrroline N-oxide (DMPO).

DNA strand breakage by hydroxyphenyl radicals generated from mutagenic diazoquinone compounds.

The mutagenic diazoquinone compounds p-diazoquinone (p-DQ), o-diazoquinone (o-DQ) and 3-diazo-N-nitrosobamethan (D-BM) cleaved the phosphodiester bond of lambda DNA, phi X174 RFI DNA and M13mp8ss DNA. p-DQ also cleaved the phosphodiester bond of bis(p-nitrophenyl)phosphate. The breakage of the phosphodiester bond was inhibited by the antioxidant butyl hydroxyanisole (BHA), ethanol, the spin trapping agent DMPO, cysteine and 2-mercaptoethanol.

Unique karyotypes in two patients with Prader-Willi syndrome.

A physical disruption of the Prader-Willi syndrome (PWS) chromosome region is thought to cause PWS. We describe 2 girls with PWS phenotype, who had unique chromosome 15 abnormalities. The first patient showed mosaicism: 45,XX,t(15;15)(qter----p11.

Changes in ovalbumin and protein synthesis in vivo in the magnum of laying hens during the egg formation cycle.

1. The present study was carried out to investigate whether or not the rate of synthesis of total protein in various oviducal segments and ovalbumin, a major egg white protein, in the magnum fluctuated during the egg formation cycle in laying hens. 2.

Synchronization culture of amniotic fluid cells using excess thymidine block followed by deoxycytidine release and its application to high-resolution banding analysis of chromosomes.

We describe a simple synchronization culture technique of amniotic fluid (AF) cells to yield many earlier mitotic divisions with extended chromosomes. AF cell samples obtained by amniocentesis were cultured in the usual manner. Thirty hours after the first subculture, they were exposed to excess thymidine (0.

Strain differences in whole-body protein turnover in the chicken embryo.

1. Whether or not there is a strain difference in embryonic whole-body protein turnover rates was tested using the chicken embryos of Rhode Island Red carrying a sex-linked dwarf gene (dwarf), White Leghorn (layer), and White Cornish X White Plymouth Rock (broiler) strains on day 12 of incubation. 2.

Importance of albumen content in whole-body protein synthesis of the chicken embryo during incubation.

The importance of egg albumen content in whole-body protein synthesis was investigated in developing chicken embryos by using lines genetically selected for high and low albumen contents and by removing albumen from eggs before incubation. 2. Whole-body protein synthesis was estimated by injecting L-[15N]-phenylalanine intravenously on day 12 of incubation.

Loss of the 3p25.3 band is critical in the manifestation of del(3p) syndrome: karyotype-phenotype correlation in cases with deficiency of the distal portion of the short arm of chromosome 3.

Two patients with monosomy for the distal portion of the short arm of chromosome 3 are described. Chromosome analysis on prometaphase cells demonstrated a karyotype of 46,XX,del(3) (p25.3) in one patient and 46,XX,r(3)(p26.

Protein synthesis in tissues and in the whole body of laying hens during egg formation.

The present study was conducted to investigate whether or not protein synthesis in tissues and in the whole body of laying hens would be affected by the position of an ovum passing along the oviduct during egg formation. Protein synthesis in tissues was measured in vivo by a primed-continuous infusion of [15N]methionine for 3 h, finishing at the time when an ovum would have stayed at one of the segments within the oviduct, i.e.

Effect of methionine and lysine deficiencies on protein synthesis in the liver and oviduct and in the whole body of laying hens.

For laying hens, protein synthesis in the liver, in the oviduct (magnum and remaining portions), and in the whole body was measured in vivo in order to investigate the effect of a dietary deficiency of methionine or lysine. The rate of protein synthesis in tissues was calculated from the incorporation of L-[15N]phenylalanine into the protein fraction; whole-body protein synthesis was estimated from the plateau enrichment of free [15N]phenylalanine in plasma. The enrichment of labeled phenylalanine was analyzed by using a gas-chromatograph mass spectrometer, following a primed infusion of the isotope for 6 h.

Superoxide dismutase-mediated reversible conversion of 3-hydroxyamino-1-methyl-5H-pyrido[4,3-b]indole, the N-hydroxy derivative of Trp-P-2, into its nitroso derivative.

Aerobic oxidation of 3-hydroxyamino-1-methyl-5H-pyrido-[4,3-b]indole [Trp-P-2(NHOH)] in neutral aqueous solution was greatly accelerated by copper-zinc superoxide dismutase (SOD). The major product in this SOD-mediated reaction was identified as 3-nitroso-1-methyl-5H-pyrido[4,3-b]indole [Trp-P-2(NO)]. This conversion was accompanied by a decrease of the mutagenicity of the mixture, as monitored by the direct-acting mutagenicity on Salmonella typhimurium TA98; a rapid change to approximately 1/3 of the original mutagenicity was followed by no further decrease of the activity.

Generation of intracellular active oxygens in mouse FM3A cells by 3-hydroxyamino-1-methyl-5H-pyrido[4,3-b]indole, the activated Trp-P-2.

Mouse FM3A cells in culture were treated with a reactive metabolite of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2), 3-hydroxyamino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2-(NHOH]. When the treated cells, which were judged as viable on the basis of trypan-blue exclusion, were subjected to nitroblue tetrazolium staining, formazan was formed inside the cells, a fact suggesting the intracellular presence of superoxide. No formazan formation was detected on treatment of the cells with Trp-P-2.