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Biomedical subjects

K Lundgren

Publications and source records attributed to K Lundgren.

At least 55 records · Page 3Linked to original sources
PubMed1989

Deoxyribonucleic acid sequence mapping on metaphase chromosomes by immunoelectron microscopy.

Nucleic acid sequences can be localized on chromosomes in the electron microscope after hybridization with a biotinylated DNA probe followed by detection with a primary antibiotin antibody and a secondary antibody coupled to colloidal gold. Hybridization probes can also be labelled with alternative ligands such as N-acetoxy-2-acetylaminofluorene (AAF), Dinitrophenyl-dUTP and Digoxigenin-dUTP. Multiple labelling is possible if these differently modified DNA probes are used in conjunction with colloidal gold preparations of varying particle sizes. A substantial signal amplification can be achieved by incubating preparations with successive cycles of primary antibiotin antibody followed by a biotinylated secondary antibody. Detection is with Streptavidin-gold, and in the case of highly and moderately repeated sequences, the signal is visible in the light microscope. Detailed protocols are given for EM in-situ hybridization to whole mount metaphase chromosomes and include instructions necessary to perform multiple sequence localization and signal amplification.

Animals↗
PubMed1989

A tuneable, narrow-band, high-intensity UV source.

More potent narrow-band UV sources need to be developed to determine the in vivo action spectra of long-term UV effects, such as photocarcinogenesis. This article describes the development of a potent, narrow-band UVB source, an Oriel solar simulator modified by the use of newly developed all-dielectric interference (ADI) filters. The sharp cut-off edges and high levels of transmission are unique features of these filters. Further, they can be produced as long-wave-pass or short-wave-pass filters with maximum transmittance at any given wavelength. The simulator is equipped with up to 4 ADI filters and potentially emits narrow UV bands. The filter combinations for narrow bands allow transmission of up to 80% of the incoming radiation. There was a homogeneous intensity area of 25 X 25 cm at a distance of 150 cm from the source in the centre of the irradiation field. The average intensity of UV available in narrow bands of UV (with a minimum half-band width of 11 nm) was 140 mW/m2. These values are sufficient to determine action spectra in groups of live animals (mice).

Equipment Design↗
PubMed1988

Role of metabolism and DNA adduct formation in the induction of sister chromatid exchanges in human lymphocytes by diethylstilbestrol.

Recent studies have shown that diethylstilbestrol (DES) induces sister chromatid exchanges (SCEs) in lymphocytes from pregnant and premenopausal women but has only a slight effect on lymphocytes from post-menopausal women or men. In this study blood specimens from premenopausal women were used to define the role of different metabolic pathways on DES-induced formation of SCEs and to determine whether conditions resulting in induction of SCEs also induced detectable levels of DNA adducts. Exposure of lymphocytes in vitro to 0-40 microM DES induced a concentration-dependent increase in SCEs. Addition of indomethacin to the cultures partially abolished DES-induced SCEs, suggesting involvement of prostaglandin synthetases in the formation of specific DES metabolites that cause SCEs. alpha-Naphthoflavone, an inhibitor of cytochrome P-450 monooxygenases, had no effect on DES-induced SCEs. Cells exposed to DES at doses sufficient to cause large increases in SCE induction did not have adducts detectable by a 32P-postlabeling assay capable of revealing adducts at a level of 1 adduct/10(9) normal nucleotides.

Benzoflavones↗
PubMed1988

Cytogenetic and chemical detection of human exposure to polyhalogenated aromatic hydrocarbons.

Peripheral lymphocytes from Taiwanese women (n = 35) exposed to polychlorinated aromatic hydrocarbons and from matched controls (n = 24) were assessed for the levels of sister chromatid exchanges (SCEs) after a 72-hour incubation of whole blood in the presence or absence of alpha-naphthoflavone (ANF) and for chromosome aberrations after 48 hours of incubation. Serum levels of polychlorinated biphenyl (PCB) congeners were measured for all individuals, and serum levels of several polychlorinated dibenzofurans (PCDFs) were measured for 12 exposed individuals by gas chromatography-mass spectometry. Blood concentrations of total PCBs in the exposed population averaged approximately 15 ppb, whereas mean PCDF values were 14 ppt. Major PCB congeners detected were 2,2' 4,4', 5,5'-hexa CB and 2,2'3,4,4',5-hexa CB. PCDFs detected were primarily 1,2,3,4,7,8,-hexachlorodibenzofuran (10.8 ppt) and 2,3,4,7,8-pentachlorodibenzofuran (2.7 ppt). Average SCE frequencies were 7.61 for controls and 7.30 for exposed individuals when assays were conducted in the absence of ANF, whereas respective values were 8.85 and 10.75 in the presence of ANF. Differences in the level of ANF-induced SCEs between the two populations were highly significant (P less than .001). Moreover, the ANF-induced SCEs were highly correlated with the serum concentrations of total PCBs and of several PCB congeners (P less than .001). Increases in ANF-induced SCEs appeared to be linear up to a PCB concentration of approximately 30 ppb. Chromosome aberration frequencies were similar in control and exposed populations. These studies demonstrate that in vivo exposure to PCBs and PCDFs result in an enhanced sensitivity of lymphocytes to the SCE-causing actions of ANF.

Benzoflavones↗
PubMed1988

Metabolic activation of alpha-naphthoflavone by 2,3,7,8-tetrachlorodibenzodioxin-induced rat liver microsomes.

Previous studies in our laboratory had demonstrated that addition of alpha-naphthoflavone (ANF) to lymphocytes from smokers or polychlorinated biphenyls (PCB)s-exposed individuals caused an increase in sister chromatid exchange (SCE) frequency whereas lymphocytes from controls were relatively unaffected. In order to investigate the mechanism responsible, metabolism of ANF by uninduced and 2,3,7,8-tetrachlorodibenzodioxin (TCDD)-induced microsomes was studied as a function of microsomal protein concentration and incubation time. Nonpolar metabolites were analyzed and the amount of conjugated (polar) and protein-bound metabolites determined. The initial ANF-metabolism rate was 10-fold higher in TCDD-induced microsomes (4.9 +/- 0.6 nmol/min per mg TCDD-induced microsomal protein vs. 0.5 +/- 0.2 nmol/min per mg uninduced microsomal protein) than in uninduced microsomes. Moreover, uninduced microsomes no longer metabolize ANF after 30-40 min while TCDD-induced microsomes metabolize ANF for longer than 2 h or until all the ANF is gone. In addition to the metabolites formed by uninduced microsomes [7,8-dihydro-7,8-dihydroxy-ANF (7,8-dihydrodiol); 5,6-dihydro-5,6-dihydroxy-ANF (5,6-dihydrodiol); 5,6-oxide-ANF and 6-hydroxy-ANF], TCDD-induced microsomes from unidentified metabolites. When TCDD-induced microsomes and 40 microM ANF were added to Chinese hamster ovary (CHO) cells, we found a correlation between the concentration of 5,6-oxide-ANF and clastogenicity to CHO cells. However, purified 5,6-oxide-ANF did not induce SCEs in CHO cells in the absence or presence of TCDD-induced microsomes. However, a minor metabolite (identified as the 9,10-dihydro-9,10-dihydroxy-ANF by acid dehydration) formed with TCDD-induced microsomes produces clastogenicity in CHO cells. These data indicate that a minor metabolite of ANF is a potent clastogen which suggests that this metabolite may be responsible for the ANF-mediated increases in SCE frequency in lymphocytes from smokers or PCB-exposed individuals.

Animals↗
PubMed1988

No detectable increase in sister chromatid exchanges in lymphocytes from mycosis fungoides patients after topical treatment with nitrogen mustard.

We measured sister chromatid exchanges (SCEs) in lymphocytes of eight MF patients before, and 2 h, and 24 h after treatment with topical nitrogen mustard. The mean number of SCEs/cell (+/- SEM) were 8.15 (+/- 0.62), 8.34 (+0.55) and 8.26 (+/- 0.56), respectively. The 2 h and 24 h results do not differ significantly from pretreatment levels. Our results show that concentrations of nitrogen mustard used to control MF do not induce SCEs in peripheral blood lymphocytes.

Aged↗
PubMed1988

Pretreatment with long-wave ultraviolet light inhibits ultraviolet-induced skin tumor development in hairless mice.

The carcinogenic effects of long-wave ultraviolet radiation (UV-A) (320 to 400 nm) irradiation followed by exposure to broad-spectrum ultraviolet (UV) irradiation were studied in 200 lightly pigmented, hairless, hr/hr C3H/Tif mice. No skin tumors were observed in the group irradiated with UV-A for four weeks (total dose, 4050 kJ/m2, observed for 57 weeks). Ultraviolet exposure induced skin tumors in a dose-dependent manner. In a group exposed to UV irradiation for 13 weeks, 35% of the mice had developed tumors after 57 weeks. Twenty-six weeks of exposure resulted in 88% of the animals being affected. In contrast it was found that treatment with UV-A irradiation (four weeks, total dose up to 4200 kJ/m2) preceding exposure to UV irradiation (13 or 26 weeks) resulted in a significantly delayed tumor development. Exposure with UV-A induced no visible changes of the skin, and subsequent microscopic examination revealed no measurable changes in epidermal thickness or melanin content. Our results suggest that, depending on the exposure schedule, UV-A in addition to previously reported carcinogenic properties also may act as an antitumor agent.

Animals↗
PubMed1987

Curvature of mouse satellite DNA and condensation of heterochromatin.

Cloned, sequenced mouse satellite DNA exhibits properties characteristic of molecules that possess a stable curvature. Circularly permuted fragments containing the region predicted to bend were used to map the curvature relative to DNA sequence. The altered mobility of these fragments in polyacrylamide gels is reversed when gels are run in the presence of distamycin A, a drug that binds preferentially to AT-rich DNA. Treatment of living mouse cells with this drug dramatically reduces the condensation of centromeric heterochromatin, the exclusive location of satellite sequences. In situ hybridization of satellite probes to extended chromosomes at the electron microscope level shows that satellite does not comprise a single block but is distributed throughout the centromere region. Based on these experiments, we hypothesize that the structure of mouse satellite DNA is an important feature of centromeric heterochromatin condensation.

Animals↗
PubMed1987

alpha-Naphthoflavone metabolized by 2,3,7,8-tetrachlorodibenzo(p)dioxin-induced rat liver microsomes: a potent clastogen in Chinese hamster ovary cells.

alpha-Naphthoflavone (ANF) is a widely used inhibitor of P-450-mediated metabolism. Previously, we have demonstrated that in vitro addition of ANF to human lymphocytes produced significantly greater numbers of sister chromatid exchanges (SCEs) in samples from smokers compared to nonsmokers. In order to study the mechanism of this differential induction, we investigated the clastogenic activity of ANF as a consequence of metabolism by induced and uninduced rat liver microsomes. Exponentially growing Chinese hamster ovary cells were treated with ANF for 2 h in the presence or absence of microsomes, followed by incubation for 12 (chromosome aberrations) or 24 h (SCEs). ANF induced concentration (4 to 40 microM)-dependent increases in SCEs and chromosome aberrations when coincubated with 2,3,7,8-tetrachlorodibenzo(p)dioxin-induced microsomes. At the lower concentrations of ANF, chromatid damage was most predominant, whereas at the higher concentrations, a high percentage of cells was killed. The surviving cells exhibited shattered chromosomes and multiple damage in the form of chromatid exchanges and breaks. ANF was not clastogenic nor did it induce SCEs in Chinese hamster ovary cells when incubated with microsomes from control rats or phenobarbital-treated rats. Moreover, NADPH was required for the clastogenic actions of ANF in the presence of 2,3,7,8-tetrachlorodibenzo(p)dioxin-induced microsomes. Analysis of the ANF metabolites by high-pressure liquid chromatography revealed that 2,3,7,8-tetrachlorodibenzo(p)dioxin-induced microsomes metabolized ANF to a much greater extent than control or phenobarbital-induced microsomes. Our results suggest that the clastogenic activity of ANF in Chinese hamster ovary cells is mediated by the cytochrome P-450 monooxygenase system.

Animals↗
PubMed1987

Effects of 5-bromo-2-deoxyuridine concentration and alpha-naphthoflavone on the association between smoking and the frequency of sister-chromatid exchanges in lymphocytes from maternal and cord blood.

The frequency of sister-chromatid exchanges was analyzed in maternal and cord blood lymphocytes obtained at delivery from 23 nonsmokers and 21 smokers. Lymphocytes were cultured under 3 conditions: in the presence of 100 microM 5-bromo-2-deoxyuridine (BUdR), 20 microM BUdR and 20 microM BUdR with 40 microM alpha-naphthoflavone (ANF). Under all assay conditions, frequencies of SCEs were consistently higher for maternal lymphocytes than for cord lymphocytes. There was no association between SCE values for cultures of the same blood specimen with 100 microM BUdR and 20 microM BUdR. When cultured with 100 microM BUdR, maternal lymphocytes from smokers had a mean SCE frequency of 13.5, which was significantly higher than the value of 11.1 observed for nonsmokers (p = 0.001 by the Wilcoxon rank sum test). Maternal smoking had no significant effect on overall frequencies of SCEs in maternal blood cultured with 20 microM BUdR either with or without ANF or when the differential between cells cultured with and without ANF was considered. Use of caffeinated beverages was associated with increased SCE values for maternal lymphocytes cultured with 20 microM BUdR (Tau beta = 0.36, p = 0.02 for the Kendall's Rank Correlation), but no such association was seen with 100 microM BUdR. For cord blood lymphocytes, however, neither smoking nor caffeine use were associated with SCE values obtained by any of the assay conditions used. The findings suggest that results of human monitoring studies using SCEs could differ depending on the concentration of BUdR used in cultures.

Benzoflavones↗
PubMed1987

Increased frequencies of alpha-naphthoflavone induced sister chromatid exchanges in long-term survivors of childhood acute lymphoblastic leukemia.

Cultures of whole-blood were established from 12 children (mean age = 13.5 years) surviving acute lymphoblastic leukemia (mean time of discontinued therapy = 4.5 years) and 10 age-matched controls were assessed for the level of baseline, alpha-naphthoflavone (ANF) and methyl-methane-sulphonate (MMS) induced sister chromatid exchanges (SCEs). The average baseline levels of SCEs in the patient group (7.65) and in the control (6.72) were different at a statistical marginal level (P = 0.05). No statistical difference was observed in the levels of MMS-induced SCEs between the two groups (P greater than 0.1). In contrast, highly significant differences (P = 0.003) were observed in the level of ANF-induced SCE-levels between the patients (10.78) and the controls (8.76).

Adolescent↗
PubMed1986

Dioxin treatment of rats results in increased in vitro induction of sister chromatid exchanges by alpha-naphthoflavone: an animal model for human exposure to halogenated aromatics.

Recent reports have shown that alpha-naphthoflavone (alpha-NF) in vivo enhances the sister chromatid exchange (SCE) frequency in lymphocytes from human populations exposed to cigarette smoke or polychlorinated biphenyls and dibenzofurans. In this study, female Sprague-Dawley rats (9-11 weeks old) were administered a single oral dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and killed 6 days after treatment. Blood cultures were established with or without alpha-NF. The baseline and alpha-NF-induced SCE frequencies were assessed in lymphocytes after a 72-hr culture period. No effect on the SCE baseline frequency (cultures without alpha-NF) was detected in rats exposed to 0-30 micrograms TCDD/kg. However, the SCE frequencies from cultures incubated in the presence of alpha-NF were significantly higher in lymphocytes from rats treated with TCDD. Moreover, delta SCE values (SCE alpha-NF minus SCE baseline) were significantly higher in lymphocytes from rats treated with TCDD than in controls. A dose-dependent increase in delta SCE values was observed between 0 and 3 micrograms TCDD/kg, followed by a plateau at higher doses. This induction pattern closely resembled the induction of the liver microsomal aryl hydrocarbon hydroxylase activity by TCDD. In contrast to TCDD, phenobarbital treatment of rats (75 mg/kg/day) had no effect on alpha-NF-induced SCE frequencies in lymphocytes. Liver microsomes from TCDD-treated rats metabolized alpha-NF at a rate much faster than that of control microsomes. These studies indicate that TCDD-exposed rats provide a useful model to investigate the mechanism of enhanced in vitro induction of SCE frequency in lymphocytes from humans exposed to toxic halogenated aromatics or cigarette smoke.

Animals↗
PubMed1986

Human monoclonal antibodies to Pf 155, a major antigen of malaria parasite Plasmodium falciparum.

Pf 155, a protein of the human malaria parasite Plasmodium falciparum, is strongly immunogenic in humans and is believed to be a prime candidate for the preparation of a vaccine. Human monoclonal antibodies to Pf 155 were obtained by cloning B cells that had been prepared from an immune donor and transformed with Epstein-Barr virus. When examined by indirect immunofluorescence, these antibodies stained the surface of infected erythrocytes, free merozoites, segmented schizonts, and gametocytes. They bound to a major polypeptide with a relative molecular weight of 155K and to two minor ones (135K and 120K), all having high affinity for human glycophorin. The antibodies strongly inhibited merozoite reinvasion in vitro, suggesting that they might be appropriate reagents for therapeutic administration in vivo.

Animals↗