Biomedical subjects

S M Rappaport

Publications and source records attributed to S M Rappaport.

94 records · Page 6Linked to original sources

PubMedDate not supplied

Application of Raney nickel to measure adducts of styrene oxide with hemoglobin and albumin.

Adducts of styrene 7,8-oxide (SO) with the blood proteins, Hb and albumin (Alb), were measured following treatment of the proteins with Raney nickel (Ra-Ni) to cleave 2-phenylethanol, which was subsequently assayed. Internal standards were prepared by modifying Hb and Alb with 4-methyl-SO to produce similar adducts. In a preliminary experiment with human blood, which had been modified with [14C]SO in vitro, it was determined that Ra-Ni released 6% of the total binding to globin and 76% of the total binding to Alb. Since Ra-Ni primarily cleaves covalently-bound sulfur species, this suggests that much more binding to human serum Alb occurred at a free cysteine residue than to human Hb. The overall rate of reaction of SO at 37 degrees C in vitro had a half-time of 0.70 h in human blood and of 0.36 h in blood from Sprague-Dawley rats. Adducts of SO with Hb and Alb were measured following SO modification of blood from humans and rats in vitro at 37 degrees C. Slopes of the linear relationships between adduct level and SO concentration allowed the following second-order rate constants [(L/mol of protein)/h] to be estimated: rat Hb, 72; human Hb, 2.4; rat Alb, 63; human Alb, 32. These rate constants show that intrinsic reactivity of SO toward the cysteine residue of human Hb is much lower than that of rat Hb or of Alb from either species. Adducts of Hb and Alb were also measured following administration of both SO and styrene to rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMedDate not supplied

Measurement of hemoglobin and albumin adducts of tetrachlorobenzoquinone.

Tetrachloro-1,4-benzoquinone (Cl4BQ), a metabolite of pentachlorophenol (PCP), is believed to play a role in the genotoxicity of PCP. We have developed a method to measure the adducts of Cl4BQ with cysteine residues of hemoglobin (Hb) and albumin (Alb). This method employs the use of Raney nickel to selectively cleave the sulfur-bound adducts. Adducts of Hb and Alb with Cl4BQ were measured following modification of rat blood with Cl4BQ (0-90 microM) in vitro. The formation of both Hb and Alb adducts was linear over the entire range with second-order rate constants of 6.89 and 167 L mol-1 h-1, respectively. The proportions of the concentrations of these Hb and Alb adducts to those of all covalently-bound products were estimated to be 0.053 and 0.178, respectively, at initial Cl4BQ concentrations between 3 and 90 microM. The overall rate of reaction of Cl4BQ in rat blood (in vitro) was pseudo-first-order with an estimated half-time of 4.35 h. Hb and Alb adducts of Cl4BQ were also measured in vivo following oral administration of PCP to rats (0-20 mg/kg body wt). Linear production of Hb and Alb adducts was observed over the entire range of dosages, with slopes of 0.09 and 8.22 pmol of adduct (g of protein)-1 [(mg of PCP)/(kg body wt)]-1, respectively. On the basis of production of Hb adducts in vitro and in vivo, it is estimated that 2.7 x 10(-7) mol of Cl4BQ was released to the blood of rats per mole of PCP administered.(ABSTRACT TRUNCATED AT 250 WORDS)

Albumins↗

PubMedDate not supplied

Characterization of chlorinated adducts of hemoglobin and albumin following administration of pentachlorophenol to rats.

Five cysteinyl adducts (including one with multiple isomeric forms) of hemoglobin (Hb) and albumin (Alb) have been characterized in the blood of Sprague-Dawley rats following administration of pentachlorophenol (PCP). Three of these adducts were formed by multiple substitution reactions of tetrachloro-1,4-benzoquinone (Cl4-1,4-BQ) and its products, and two arose from reactions of tetrachloro-1,4-benzosemiquinone (Cl4-1,4-SQ) and tetrachloro-1,2-benzosemiquinone (Cl4-1,2-SQ). Adducts of tetrachloro-1,2-benzoquinone (Cl4-1,2-BQ) were not observed. Regarding adducts of Cl4-1,4-BQ and its products, specific structures were assigned to monosubstituted, disubstituted, and trisubstituted adducts of Hb and Alb following modification of rat blood with Cl4-1,4-BQ (0-45 microM) in vitro and after metabolism of PCP (0-40 mg/kg body weight) in Sprague-Dawley rats, in vivo. The formation of all adducts was linear over the ranges tested, with Alb adducts being more abundant than Hb adducts. The levels of the adducts measured were in the following order: monosubstituted > disubstituted > trisubstituted. The observation that Cl4-1,4-BQ can produce multisubstituted adducts with proteins suggests that protein-protein cross links may be formed, with inherent toxicological implications. Regarding adducts of the semiquinones (detected only in vivo), linear production of Hb and Alb adducts was observed with increasing dosage of PCP for adducts of both Cl4-1,4-SQ and Cl4-1,2-SQ. Higher levels of the semiquinone adducts were observed in Hb than in Alb, in contrast to the results with the quinone adducts. In a separate in vivo experiment (20 mg PCP/kg body weight), where animals were sacrificed at intervals up to 336 h postadministration, adducts were eliminated at rates which were comparable among the different adducts of a given protein.

Administration, Oral↗

PubMedDate not supplied

Variability of albumin adducts of 1,4-benzoquinone, a toxic metabolite of benzene, in human volunteers.

A putative haematotoxic and leukaemogenic metabolite of benzene, 1,4-benzoquinone (1,4-BQ), reacts rapidly with macromolecules. The authors previously characterized levels of the albumin (Alb) adduct (1,4-BQ-Alb) of this reactive species in populations of workers exposed to benzene. Since high levels of 1,4-BQ-Alb were also measured in unexposed workers from those investigations, the current study was initiated to determine potential sources of 1,4-BQ in the general population. A single blood sample was collected from 191 healthy subjects from the Research Triangle area, NC, USA, to determine the baseline 1,4-BQ-Alb levels and contributing sources. The median 1,4-BQ-Alb at baseline was 550?pmol?g(-1) Alb (interquartile range 435-814?pmol?g(-1)). A second blood sample was collected approximately 3 months later from a subgroup of 33 subjects to estimate the within- and between-person variation in 1,4-BQ-Alb. Standardized questionnaires were administered to collect information about demographic, dietary and lifestyle factors. Multiple linear regression models identified several significant contributors to 1,4-BQ-Alb levels, including gender, body mass index (BMI), the gender-BMI interaction, automobile refuelling, smoking status, and consumption of fruit and the artificial sweetener, aspartame. The authors predicted that these background levels of 1,4-BQ-Alb were equivalent to occupational exposures between 1 and 3 parts per million of benzene. Mixed effects linear models indicated that the random variation in adduct levels was about equally divided between and within subjects. The observations indicate that levels of 1,4-BQ-Alb cover a wide range in the general population, and they support the hypotheses that demographic, diet and lifestyle factors are contributing sources.

Adult↗