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Biomedical subjects

K Hemminki

Publications and source records attributed to K Hemminki.

At least 307 records · Page 17Linked to original sources
PubMed1991

32P-postlabeling of N-7, N2 and O6 2'-deoxyguanosine 3'-monophosphate adducts of styrene oxide.

Adducts were prepared by reacting styrene oxide with 2-deoxyguanosine 3'-monophosphate (dGMP). Four isomeric N-7-, two diastereomeric N2- and three isomeric O6-adduct were isolated and characterized. The adducts were used as substrates in the 32P-postlabeling reaction. No phosphorylation products were seen with the N-7-alkylation products. One diastereomeric N2-adduct was labeled with 20% efficiency and the second with a markedly lower efficiency. Two of the three O6-adducts were labeled with 5% and the third with 10% labeling efficiency. The results suggest that large N-7-dGMP adducts are very poor substrates of T4 polynucleotide kinase. The diastereomeric products are labeled at different efficiencies indicating stereoselectivity in the kinase reaction.

Deoxyguanine Nucleotides↗
PubMed1991

Quantitation of the 32P-postlabeling reaction using cyclic N1,N2 and C8 modified deoxyguanosine 3'-monophosphates as substrates.

The 32P-postlabeling technique was used to investigate the efficiency of phosphorylation reaction by T4 polynucleotide kinase using seven synthetic adducted deoxyguanosine 3'-monophosphates. The adducts included cyclic N1,N2 derivatives and the C8 adduct of 4-aminobiphenyl. The adducted substrates were detected at a subfemtomole sensitivity except for one of the diastereomeric propanoguanine derivatives. In general, the recommended conditions were found to be proper for an efficient phosphorylation of the adducts studied. Sensitivity of the adducts to the 3'-dephosphorylation reaction of nuclease P1 was also tested. All the complex cyclic adducts were resistant towards P1. However, the ethenoguanine and 4-aminobiphenyl adducts were relatively sensitive towards P1. No differences were noted between diastereomers.

Chromatography, Thin Layer↗
PubMed1991

Quantitative and kinetic examination of 32P-postlabeling of etheno-substituted nucleotides.

1,N6-ethenodeoxyadenosine-, 1,N2-ethenodeoxyguanosine- and 3,N4-ethenodeoxycytidine-3'-monophosphates were labeled by [gamma-32P] ATP using T4 polynucleotide kinase in conditions commonly used for the 32P-postlabeling assay. Kinetic studies showed that the reaction is fast reaching a plateau after 15-30 min. The efficiency of phosphorylation, as studied by substrate-product concentration dependency, was between 50-100% at the lower substrate concentrations. The adducts are labeled efficiently at sub-femtomole levels. All the adducts were sensitive to the 3'-dephosphorylation by P1 nuclease although the guanine derivative appeared to be more resistant than the two other adducts.

Adenosine Triphosphate↗
PubMed1991

Assay for nucleoside and nucleotide binding of a potent mutagen, 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone.

Deoxyadenosine, deoxyguanosine, deoxycytidine; thymidine and a dinucleotide, 2'-deoxycytidylyl-(3'-5')-2'-deoxythymidine (dCdT), were reacted with 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX). Nucleoside HPLC analysis revealed that the major products are the degradation products of MX and no stable MX-adducts were found. The same result was obtained with sodium borohydride reduction. The fluorescence spectra of deoxyguanosine-MX showed no fluorescence at neutral or alkaline pH, indicating no binding products to the N2-groups of deoxyguanosine.

Borohydrides↗
PubMed1991

Postlabeling analysis of polycyclic aromatic hydrocarbon-DNA adducts in white blood cells of foundry workers.

Blood samples were obtained from 61 volunteers working in a Finnish iron foundry who were exposed to carcinogenic polycyclic aromatic hydrocarbons and from 19 control subjects not known to be exposed to these chemicals. Foundry workers were categorized into high, medium, or low exposure groups, based on their exposure levels to airborne benzo[a]pyrene (BaP) (high greater than 0.2, medium 0.05-0.2, low less than 0.05 micrograms BaP/m3 air). Aromatic adducts were determined in white blood cell DNA from exposed and unexposed subjects using 32P-postlabeling. There was a highly significant correlation between the estimated exposure and adduct levels as determined by analysis of variance. The levels of adducts found in the high and medium group samples ranged from 5 to over 20 adducts in 10(8) DNA nucleotides. No effects due to age, sex, or the smoking habits of the subjects were observed. Within each group, the SEM appeared rather small. This study demonstrates the utility of the 32P-postlabeling assay to assess human exposure to known and unknown environmental aromatic genotoxicants.

Adult↗
PubMed1991

DNA adducts in lymphocytes and granulocytes of smokers and nonsmokers detected by the 32P-postlabelling assay.

The effects of smoking and DNA adduct formation were analysed in isolated human white blood cell populations. As the white cells are composed mainly of granulocytes with a short half-life and T-lymphocytes with a half-life of several years, we isolated the lymphocytes and granulocytes of 11 smokers and 10 nonsmokers to determine any smoking-related DNA adducts by the nuclease-P1-enhanced 32P-postlabelling assay. The differences between the mean lymphocyte DNA adducts/10(8) nucleotides of 31 +/- 5.7 (SE) of smokers were significantly higher (P less than 0.05) than those in the lymphocytes 13 +/- 1.6 (SE) of nonsmokers. The total DNA adducts/10(8) nucleotides obtained from the granulocytes of smokers and nonsmokers was 9.6 +/- 1.9 and 7.6 +/- 1.9 respectively. The plasma cotinine concentrations were in good agreement with the smoking information given by the individual smokers (r = 0.847, P less than 0.001). The DNA adduct levels of the lymphocytes of the 10 smokers correlated with the plasma cotinine concentrations (r = 0.639, P less than 0.05). The variation between the results was explained by the variation among the individuals and the samples, but not by the variation in the parallel determinations. More detailed studies are needed to analyse the source of the individual variations between the smokers' adduct levels, DNA repair, and differences in the metabolism of the compounds in cigarette smoke.

Adult↗
PubMed1991

Measurement by 32P-postlabelling of 7-methylguanine levels in white blood cell DNA of healthy individuals and cancer patients treated with dacarbazine and procarbazine. Human data and method development for 7-alkylguanines.

A 32P-postlabelling method was developed to measure 7-methylguanine in human DNA. DNA was digested to nucleotides and 7-methyl-2'-deoxyguanosine-3'-monophosphate (7-me-dGMP) was isolated from normal nucleotides using strong anion-exchange column chromatography. Overall the method gave 35-45% yield as measured with DNA methylated with tritiated dimethyl sulfate. Total white blood cell DNA from healthy non-smokers (n = 17) contained from 2.5 7-methylguanine residues/10(7) nucleotides, corrected for the losses in preparation. Among four patients sampled immediately after a total dose of 1050-2800 mg of dacarbazine or procarbazine, the mean adduct level was 57 7-methylguanine residues/10(7) nucleotides. As further method development, we also investigated the phosphorylation reaction by T4 polynucleotide kinase using dinucleotides containing 7-methylguanine and corresponding imidazole ring-opened products as substrates. We found that imidazole ring-opened dTpdG-Me is resistant to digestion with deoxyribonuclease I, snake venom phosphodiesterase and prostatic acid phosphatase. It is quantitatively phosphorylated at femtomolar levels. This method is shown to be suitable for the detection of 7-methylguanine in DNA, and is suggested to be the approach most suited to postlabelling large and labile 7-alkylguanines in DNA.

Chromatography, Thin Layer↗
PubMed1991

Effects of paternal occupational exposure on spontaneous abortions.

BACKGROUND: Paternal exposure to mutagenic agents has been suggested to affect pregnancy outcome adversely. METHODS: A nationwide data base of medically diagnosed spontaneous abortions and other pregnancies and national census data was used to evaluate the effects of men's occupational exposures on risk of spontaneous abortion in 99,186 pregnancies in Finland. Census data from the years 1975 and 1980 provided information about the occupation, industry, and socioeconomic status. A job-exposure classification was developed to classify women and their husbands according to possible occupational exposures on the basis of their occupational title and industry. RESULTS: In 10% of the pregnancies, the husband was exposed to one or more of the mutagens, and the rate of spontaneous abortion was unaffected (OR = 1.0). Of the 25 specific mutagenic exposures evaluated, paternal exposure to four (ethylene oxide, rubber chemicals, solvents used in refineries, and solvents used in the manufacturing of rubber products) was associated with an increased relative risk of spontaneous abortion. In addition, the risk of spontaneous abortion was higher among wives of rubber products workers than among unexposed men. CONCLUSIONS: Although there is some biological rationale for the findings of this study, these findings need to be confirmed by studies in which individual exposures can be measured directly.

Abortion, Spontaneous↗
PubMed1991

Immunologic measurement of polycyclic aromatic hydrocarbon-albumin adducts in foundry workers and roofers.

Roofers and iron foundry workers with high exposure to polycyclic aromatic hydrocarbons (PAH) were monitored for levels of covalent PAH serum albumin adducts, quantitated in enzymatically digested samples by a sensitive competitive enzyme-linked immunosorbent assay. Albumin adducts were higher in the foundry workers (5.22 fmol/micrograms, 0.314 mmol/mol) than in the reference group (4.07 fmol/micrograms, 0.245 mmol/mol), but only of borderline significance probably due to the small sample size. In a subset of foundry workers, a significant difference in adduct levels was observed for samples collected immediately after vacation and after six weeks of workplace exposure. The roofers also showed higher levels of adducts (5.19 fmol/micrograms, 0.312 mmol/mol) than their reference group (3.28 fmol/micrograms, 0.197 mmol/mol). These results demonstrate the feasibility of PAH protein adduct measurement as a marker of human exposure to this class of chemicals.

Antibodies, Monoclonal↗
PubMed1991

Paternal occupational lead exposure and spontaneous abortion.

A case-referent study was conducted on whether occupational exposure of men to inorganic lead is related to spontaneous abortion in their wives. The men were biologically monitored for lead exposure. The cases (213 spontaneous abortions) and referents (300 births) were identified from medical registers. Lead exposure was assessed with blood lead (PbB) measurements and data obtained from a questionnaire. The results did not show a statistically significant relationship between spontaneous abortion and paternal lead exposure among all the study subjects. A significant increase was observed in the risk of those women whose husbands had been monitored (PbB greater than or equal to 1.5 mumol/l) during or close to the time of spermatogenesis. The association between lead exposure and spontaneous abortion was modified also by the age of the wife and paternal alcohol use. The findings suggest that there may be an association between paternal lead exposure and the risk of spontaneous abortion.

Abortion, Spontaneous↗
PubMed1991

DNA adducts and other biological markers in risk assessment for environmental carcinogens.

In order to validate a number of biologic markers as indicators of effective dose or potential risk, we have applied them in related cross-sectional, longitudinal and case-control studies. Biologic markers such as carcinogen-DNA adducts and activated oncogenes have already provided insights into mechanisms of chemical carcinogenesis. They have also given information regarding the extent of interindividual variation in biologic response within the human population. These results are directly relevant to quantitative risk assessment. However, more research is needed to establish quantitative links between biologic markers and human risk of cancer.

Biomarkers↗
PubMed1991

Use of exposure information and animal cancer data in the prevention of environmental and occupational cancer.

In order to prevent cancer, a hazard should optimally be identified before humans are exposed to it. Therefore, valid procedures should be available for predicting human risks on the basis of experimental results. The adequacy of experimental data for identifying potential human health risks have been the subject of scientific question and debate. Use of such results and of data on exposure in setting priorities for cancer prevention should, in principle, be feasible in view of the relatively good correlation between animal and human data. Studies have also revealed a reasonable quantitative relationship between carcinogenic potencies of several chemicals in laboratory rodents and humans. The ranking of cancer hazards on the basis of experimental data requires estimates of both the extent of human exposure and of the size of the exposed population. Occupational exposures can result in high individual exposures, whereas the general population is usually exposed to lower levels. Action for cancer prevention can be taken at both levels.

Animals↗
PubMed1990

Spontaneous abortions among women exposed to organic solvents.

We investigated the association between medically diagnosed spontaneous abortions and occupational exposure to organic solvents. The study population was composed of women biologically monitored for solvents. The workers were classified into exposure categories on the basis of work description and the use of solvents as reported in the questionnaires, and on biological exposure measurements. The odds ratio of spontaneous abortion for solvent exposure, adjusted for potentially confounding factors, was significantly increased (2.2, 95% confidence interval [CI] = 1.2-4.1). The increase was most consistent among workers exposed to aliphatic hydrocarbons: the odds ratio for high exposure was 3.9 (95% CI = 1.1-14.2) and for exposed graphics workers 5.2 (95% CI = 1.3-20.8). The odds ratio was increased also among toluene-exposed shoe workers (9.3, 95% CI = 1.0-84.7). The results of the study support the hypothesis of a positive association between spontaneous abortion and exposure to organic solvents during pregnancy and suggest that exposure, especially to aliphatic hydrocarbons, increases the risk of abortion.

Abortion, Spontaneous↗
PubMed1990

32P-postlabeling of 7-methylguanine adducts in DNA.

The 32P-postlabeling technique introduced by Randerath and co-workers has been particularly successful with stable and nonpolar DNA adducts, but the assay has not been used to any large extent to detect 7-alkylguanine derivatives. In the present communication, we have investigated the phosphorylation reaction by T4 polynucleotide kinase using 7-methyl-3'-dGMP, ring-opened 7-methyl-3'-dGMP and enzyme-digested methylated DNA as substrates. The methylated substrates were detected at femtomol (fmol) sensitivities. 7-methyl-3'-dGMP was quantitatively phosphorylated at these low concentrations. The efficiency of phosphorylation of the ring-opened product was less. It was shown that ring-opened 7-methyl-3'-dGMP was resistant to digestion with nuclease P1, making alkali-treatment and enzyme digestion of DNA possible approaches to the determination of 7-methylguanine in DNA.

Animals↗
PubMed1990

Aromatic DNA adducts in white blood cells of coke workers.

White blood cell DNA adducts were measured in coke workers, local controls and countryside controls using the 32P-postlabelling technique. The method detected aromatic adducts including those formed by polycyclic aromatic hydrocarbons (PAHs). Coke workers are heavily exposed to PAHs particularly when working at the batteries. A difference in adduct levels was noted between the coke workers at the battery as compared to other jobs. The adduct levels in the non-battery were higher than those in the countryside controls.

Air Pollutants, Occupational↗
PubMed1990

Protein binding, sister chromatid exchange and expression of oncogene proteins in patients treated with cisplatinum (cisDDP)-based chemotherapy.

The aim of this pilot was to evaluate the feasibility of incorporating several complementary biologic markers into a molecular epidemiologic study of chemotherapy patients. Thirty-two cancer patients being treated with cis-DDP-based chemotherapy for the first time were enrolled in the study and donated a baseline sample and at least one post-treatment sample of blood. Sister Chromatid Exchange (SCEs) and plasma protein and hemoglobin binding by cisDDP were significantly increased in samples drawn at various timepoints following treatment. The pattern of nine different oncogene protein products (including those of ras, fes, and myc) remained unchanged in sera of six patients followed over the course of their treatment. However, the levels of ras P21 product were significantly elevated above normal, control levels in all six cancer patients--both prior to and throughout the course of chemotherapy. These results suggest the usefulness of utilizing a battery of markers to evaluate biologic response to cisplatinum-based chemotherapy.

Aged↗