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R Lipman

Publications and source records attributed to R Lipman.

32 records · Page 2Linked to original sources
PubMed1987

Reaction of acid-activated mitomycin C with calf thymus DNA and model guanines: elucidation of the base-catalyzed degradation of N7-alkylguanine nucleosides.

Mitomycin C (MC, 1) forms covalent adducts under acidic activating conditions (pH approximately 4) with deoxyguanosine, d(GpC), and guanine residues of calf thymus DNA. In the case of deoxyguanosine, five adducts arise from a common precursor, N7-(2'' beta, 7''-diaminomitosen-1''-yl)-2'-deoxyguanosine (10a; not isolated), which hydrolyzes spontaneously via two pathways: scission of the glycosidic bond to form N7-(2'' beta, 7''-diaminomitosen-1'' alpha-yl)guanine (5) and its 1'' beta-isomer (6) and imidazolium ring opening to generate three 2,6-diamino-4-hydroxy-5-(N-formyl-2'' beta, 7''-diaminomitosen-1'' beta-yl)pyrimidine (FAPyr) derivatives that are substituted at N6 by isomeric 2'-deoxyribose units [i.e., 1' beta-furanose (7), 1' alpha-furanose (8), and 1' beta-pyranose (9)]. The structures of 5-9 were determined by spectroscopic methods. The same five adducts were obtained from d(GpC), but only the guanine adducts 5 and 6 were formed in DNA. Adducts 7-9 interconvert during high-performance liquid chromatography (HPLC). The unexpected isomerization of the deoxyribose moiety of the initially formed 1' beta-furanose adduct 7 to those of 8 and 9 occurs upon imidazolium ring opening, as discerned by the course of imidazolium cleavage of the simple models N7-ethyl- and N7-methylguanosine and N7-methyl-2'-deoxyguanosine. All ring-opened N7-alkylguanosine derivatives studied here exist as a mixture of distinct N-formyl rotamers, manifested by multiple interconverting peaks on HPLC and in the 1H NMR spectra. In the UV spectra of such derivatives, a new and diagnostic maximum at 218 nm (at pH 7) is observed. Acid-activated MC is found to alkylate preferentially the Gua-N7 position in deoxyguanosine or d(GpC), in contrast to reductively activated MC, which preferentially alkylates the Gua-N2 position. This finding is explained by the different electronic structures of acid- and reduction-activated MC. In DNA, the N7 specificity of acid-activated MC is partially offset by steric factors.

Chemical Phenomena↗
PubMed1987

Isolation and structure of a covalent cross-link adduct between mitomycin C and DNA.

A DNA cross-link adduct of the antitumor agent mitomycin C (MC) to DNA has been isolated and characterized; the results provide direct proof for bifunctional alkylation of DNA by MC. Exposure of MC to Micrococcus luteus DNA under reductive conditions and subsequent nuclease digestion yielded adducts formed between MC and deoxyguanosine residues. In addition to the two known monoadducts, a bisadduct was obtained. Reductive MC activation with Na2S2O4 (sodium dithionite) leads to exclusive bifunctional alkylation. The structure of the bisadduct was determined by spectroscopic methods that included proton magnetic resonance, differential Fourier transform infrared spectroscopy, and circular dichroism. Formation of the same bisadduct in vivo was demonstrated upon injection of rats with MC. Computer-generated models of the bisadduct that was incorporated into the center of the duplex B-DNA decamer d(CGTACGTACG)2 indicated that the bisadduct fit snugly into the minor groove with minimal distortion of DNA structure. A mechanistic analysis of the factors that govern monofunctional and bifunctional adduct formation is presented.

Chemical Phenomena↗
PubMed1986

Reassignment of the guanine-binding mode of reduced mitomycin C.

Mitomycin C (1) is a clinically used antitumor antibiotic that binds covalently to deoxyribonucleic acid under reductive or acidic catalysis. We have determined the structures of the adducts resulting from attack of reductively activated 1 on the dinucleoside phosphate d(GpC) to be N2-(2'' beta, 7''-diaminomitosen-1''alpha-yl)-2'-deoxyguanosine (2) and its 1'' beta-isomer (3). This represents a revision of the previously reported structures for these adducts in that the mitomycin residue is linked to the N2- rather than O6-position of 2'-deoxyguanosine. This revision is the result of applying to the mitomycin case a newly developed general method that leads to unambiguous assignment of the linkage position in complex alkylated guanosines. The method as described here takes advantage of the resolution enhancement gained by calculation of the second derivatives of absorbance Fourier transform infrared spectra. In addition, we present 1H NMR data that corroborate the assigned structures of 2 and 3 and that should serve as a useful reference for future investigations into the binding of mitomycin C to DNA. The convenient synthesis of adducts 2 and 3 from deoxyguanosine and mitomycin C reported here should facilitate such investigations as well. Furthermore, we demonstrate a useful acetylation procedure for adducts and metabolites of mitomycin C that furnishes spectroscopically superior chemical derivatives (e.g., triacetates 4 and 5, derived from acetylation of adducts 2 and 3).

Deoxyguanosine↗
PubMed1986

Reaction of DNA with chemically or enzymatically activated mitomycin C: isolation and structure of the major covalent adduct.

The antitumor antibiotic mitomycin C is shown to form a covalent complex with calf thymus DNA under anaerobic conditions in the presence of either NADPH cytochrome c reductase/NADPH, xanthine oxidase/NADH, or the chemical reducing system H2/PtO2. Digestion of the complex with DNase I/snake venom diesterase/alkaline phosphatase yields a single mitomycin deoxyguanosine adduct as the major DNA alkylation product, identified as N2-(2'' beta,7''-diaminomitosen-1'' alpha-yl) 2'-deoxyguanosine (Structure 2). Two minor adducts, 2-5% each of the total adduct pool, are isolated and identified as the 1'' beta stereoisomer of 2 (Structure 3), and 10''-decarbamoyl-2 (Structure 7). The same results were obtained with M13 DNA and poly(dG-dC).poly(dG-dC); however, in the latter case, a minor adduct apparently possessing two deoxyguanosine and one mitomycin unit is isolated. Digestion of the covalent mitomycin-calf thymus DNA complex with nuclease P1 yields four dinucleotide adducts, all of which consist of 2 linked at its 3' end to each of the four possible 5' nucleotides (A, T, G, and C). Upon treatment of each dinucleotide adduct with snake venom diesterase/alkaline phosphatase, 2 is released along with the corresponding free nucleoside. In apparent conflict with the present results, previous reports from another laboratory have indicated that modification of calf thymus DNA by mitomycin C under conditions identical to those described here result in the isolation of three mitomycin C mononucleotide adducts possessing linkages of the drug to N2 and O6 of guanine and N6 of adenine. Evidence is shown suggesting that the latter adducts are actually three of the above four dinucleotide derivatives of 2 obtained independently by us and, thus, all of them in fact possess an identical N2-mitosenylguanine adduct moiety. Model-building studies indicate an excellent fit of the guanine N2-linked drug molecule inside the minor groove of B-DNA with no appreciable distortion of the DNA structure.

Alkaline Phosphatase↗
PubMed1983

Plasma immunoreactive beta-endorphin levels in depression. Effect of electroconvulsive therapy.

Immunoreactive (ir) plasma beta-endorphin level was assayed in ten symptomatic patients with a unipolar major depressive disorder and in 16 psychiatrically normal controls matched for age and sex. Plasma ir-beta-endorphin level in depressed patients was similar to that in controls. All depressed patients was similar to that in controls. All depressed patients had a transient, approximately threefold increase in ir-beta-endorphin after each use of electroconvulsive therapy (ECT). The increase of plasma ir-beta-endorphin level after ECT parallels the transient elevation of adrenocorticotropic hormone level reported by others and probably reflects a hypothalamic response to ECT.

Adrenocorticotropic Hormone↗
PubMed1981

Reductive metabolism and alkylating activity of mitomycin C induced by rat liver microsomes.

Mitomycin C, an antitumor antibiotic, is rapidly metabolized in the presence of rat liver microsomes. NADPH and anaerobic conditions are required for the process. The products isolated after reexposure to air are 2,7-diaminomitosene derivatives. Specifically, in the presence of inorganic phosphate, 1,2-cis- and -trans-2,7-diaminomitosene 1-phosphates, 1,2-cis- and -trans-2,7-diamino-1-hydroxymitosenes, and 2,7-diaminomitosene are formed. The last substance is a new mitomycin C derivative, and proof for its structure is presented. Mytomycin C has been previously postulated to be an alklating agent requiring reduction for activity (Iyer, V. N., & Szybalski, W. (1964) Science (Washington, D.C.) 145, 55]. The 1-phosphates above represent the first chemically characterized bioreductive alkylation products of the drug. 5'-Uridylic acid is alkylated analogously under these conditions, to give cis- and trans-2,7-diaminomitosene 1-(5'-uridylate), while the phosphodiester UpU and uridine itself are inert. Hydrogen gas/PtO2 gives the same results as microsomes/NADPH. The formation of the observed compounds indicates that enzymatic (or chemical) reduction of the quinone system of mitomycin C induces ring opening of the aziridine function, generating a reactive center at the C1 position as previously postulated by others (ibid.). The second alkylating center, also postulated, is not evident, however, under the conditions tested, indicating that the aziridine is the primary bioreductive alkylation function of mitomycin C. Identification of the products and mechanism of the microsomal anaerobic metabolism of mitomycin C are significant in view of the reported toxicity of the drug to anaerobic cancer cells.

Alkylation↗
PubMed1979

Phasing down state hospitals: integrated versus nonintegrated services.

Two mental health catchment areas in Massachusetts that were in the process of phasing down state hospitals and building up community care provided an opportunity for studying the effectiveness of an integrated service delivery model and a nonintegrated model. The area that integrated state hospital and community services was more successful in the phase-down. It had a lower admission rate, the patients who were readmitted did not stay as long, and patients discharged spent more time in the community. Specific adminstrative and clinical structures that facilitated the patients' progress included investing community-based, administrative authority in one person; having a centralized intake and referral system; and using case managers to follow the clients through the service delivery system.

Catchment Area, Health↗
PubMed1978

Electrostatic complexes of mitomycin C with nucleic acids and polyanions.

Reductively activated mitomycin C exhibits strong, non-covalent electrostatic binding to polyanions such as polyvinylsulfate and polyphosphate. The protonated C-2 amino group generated by the reduction is most likely responsible for this type of interaction. At moderate drug and salt concentrations only covalent binding to nucleic acids is observable. This is shown to be guanine-specific in DNA for the first time, as well as in synthetic polyribo- and polydeoxyribonucleotides at 10--20 times higher binding levels than previously tested. At higher mitomycin C concentration, however, strong non-covalent electrostatic binding to nucleic acids also occurs, resulting in a binding ratio up to 1 mol drug bound per mol mononucleotide, although this non-specific binding is relatively inhibited compared to polyvinylsulfate. Salts also have an inhibitory effect on the non-specific binding to nucleic acids. A series of mitomycin derivatives were compared for their binding and cross-linking abilities using DNA as substrate, with the following results: (a) the presence of a basic nitrogen . funtion at C-2 promotes binding, both covalent and electrostatic, presumably by kinetically facilitating the approach between positively charged nitrogen and DNA. (b) The aziridine ring is the major covalent binding site, indispensable for crosslinking and determines the guanine-specificity of the binding.

Anions↗
PubMedDate not supplied

Calorie restriction increases light-dependent photoreceptor cell loss in the neural retina of fischer 344 rats.

We investigated the effect of > or = 8 months of 40% caloric restriction (CR) on photoreceptor cell loss in 12, 18, and 24 month-old Fischer 344 rats (N = 154). Rats were reared at the NIA Biomarkers Program, National Center for Toxicological Research. Photoreceptor cell density, assessed histologically, declined with age in both the CR-fed and ad lib (AL)-fed cohorts (P < 0.000), but declines were more pronounced in the CR cohort (P < 0.0005). The deleterious effect of CR was most pronounced in the central as opposed to the peripheral retina (P = 0.008), suggesting a light-dependent mechanism. Photoreceptor cell density was inversely associated with rearing under bright light (300-750 lux) as compared with rearing under lower illuminance (< or = 200 lux) (P < 0.0005). However, the deleterious effect of bright light on photoreceptor cell density was more pronounced in the CR cohort (P = 0.04). Effects of CR on circadian activity are likely to increase the actual light exposure of the CR cohort and may explain the apparent inability of CR to delay retinal aging in albino rats.

Aging↗