Biomedical subjects

W Deng

Publications and source records attributed to W Deng.

At least 37 records · Page 2Linked to original sources

PubMed2001

Fluorescence in situ hybridization of metaphase chromosomes in suspension.

PURPOSE: To describe a new method for FISH analysis of metaphase chromosomes in suspension. MATERIALS AND METHODS: Metaphase chromosomes in suspension were isolated from a Chinese hamster human hybrid cell line, 314-2 (1) Y, and a human cell line, GM 130B. During suspension hybridization, specific chromosomes were labeled from these two cell lines using either biotin-labeled human genomic DNA, a directly labeled human pancentromere DNA probe or a chromosome #1 locus-specific probe. RESULTS: The method allows, for the first time, recovery of large numbers of isolated individual hybridized chromosomes with good morphology for both human x hamster hybrid and human cell lines. The results showed that 46-73% of the starting number of total chromosomes can be recovered after a FISH in suspension procedure. The well-preserved morphology of hybridized metaphase chromosomes allowed (1) rapid detection of individual human and hamster chromosome aberrations, (2) rapid counting of the painted human chromosomes and (3) fast, clear detection of chromosome region-specific probes. This method offers a new tool to assay chromosomes and DNA: it offers the possibility to develop new techniques for sorting chromosomes based on FISH signals, for early detection and screening of genetic diseases and for bulk measurement of both balanced or unbalanced chromosomal exchanges and rearrangements. CONCLUSION: The potential of the method described should facilitate fast, sensitive population monitoring, and increase sensitivity of the measurements in chromosome-based biodosimetry.

Animals↗

PubMed2001

Locus of enterocyte effacement from Citrobacter rodentium: sequence analysis and evidence for horizontal transfer among attaching and effacing pathogens.

The family of attaching and effacing (A/E) bacterial pathogens, which includes diarrheagenic enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC), remains a significant threat to human and animal health. These bacteria intimately attach to host intestinal cells, causing the effacement of brush border microvilli. The genes responsible for this phenotype are encoded in a pathogenicity island called the locus of enterocyte effacement (LEE). Citrobacter rodentium is the only known murine A/E pathogen and serves as a small animal model for EPEC and EHEC infections. Here we report the full DNA sequence of C. rodentium LEE and provide a comparative analysis with the published LEEs from EPEC, EHEC, and the rabbit diarrheagenic E. coli strain RDEC-1. Although C. rodentium LEE shows high similarities throughout the entire sequence and shares all 41 open reading frames with the LEE from EPEC, EHEC, and RDEC-1, it is unique in its location of the rorf1 and rorf2/espG genes and the presence of several insertion sequences (IS) and IS remnants. The LEE of EPEC and EHEC is inserted into the selC tRNA gene. In contrast, the Citrobacter LEE is flanked on one side by an operon encoding an ABC transport system, and an IS element and sequences homologous to Shigella plasmid R100 and EHEC pO157 flank the other. The presence of plasmid sequences next to C. rodentium LEE suggests that the prototype LEE resided on a horizontally transferable plasmid. Additional sequence analysis reveals that the 3-kb plasmid in C. rodentium is nearly identical to p9705 in EHEC O157:H7, suggesting that horizontal plasmid transfer among A/E pathogens has occurred. Our results indicate that the LEE has been acquired by C. rodentium and A/E E. coli strains independently during evolution.

Adhesins, Bacterial↗

PubMed2001

Characterization of mouse Atp6i gene, the gene promoter, and the gene expression.

Solubilization of bone mineral by osteoclasts depends on the formation of an acidic extracellular compartment through the action of a V-type ATPase. We previously cloned a gene encoding a putative osteoclast-specific proton pump subunit, termed OC-116 kDa, approved mouse Atp6i (ATPase, H+ transporting, [vacuolar proton pump] member I). The function of Atp6i as osteoclast-specific proton pump subunit was confirmed in our mouse knockout study. However, the transcription regulation of Atp6i remains largely unknown. In this study, the gene encoding mouse Atp6i and the promoter have been isolated and completely sequenced. In addition, the temporal and spatial expressions of Atp6i have been characterized. Intrachromosomal mapping studies revealed that the gene contains 20 exons and 19 introns spanning approximately 11 kilobases (kb) of genomic DNA. Alignment of the mouse Atp6i gene exon sequence and predicted amino acid sequence to that of the human reveals a strong homology at both the nucleotide (82%) and the amino acid (80%) levels. Primer extension assay indicates that there is one transcription start site at 48 base pairs (bp) upstream of the initiator Met codon. Analysis of 4 kb of the putative promoter region indicates that this gene lacks canonical TATA and CAAT boxes and contains multiple putative transcription regulatory elements. Northern blot analysis of RNAs from a number of mouse tissues reveals that Atp6i is expressed predominantly in osteoclasts, and this predominant expression was confirmed by reverse-transcription polymerase chain reaction (RT-PCR) assay and immunohistochemical analysis. Whole-mount in situ hybridization shows that Atp6i expression is detected initially in the headfold region and posterior region in the somite stage of mouse embryonic development (E8.5) and becomes progressively restricted to anterior regions and the limb bud by E9.5. The expression level of Atp6i is largely reduced after E10.5. This is the first report of the characterizations of Atp6i gene, its promoter, and its gene expression patterns during mouse development. This study may provide valuable insights into the function of Atp6i, its osteoclast-selective expression, regulation, and the molecular mechanisms responsible for osteoclast activation.

Adenosine Triphosphatases↗

PubMed2001

A study to verify a reported excess of chromosomal aberrations in blood lymphocytes of Namibian uranium miners.

This report describes a study to verify an earlier report of excess chromosomal damage in the blood lymphocytes of uranium miners. Coded blood samples from 10 miners and 10 controls were analyzed conventionally for unstable aberrations and by FISH for translocations. Conventional analysis, scoring 1000 metaphases per subject, showed no significant difference between miners and controls in the frequencies of chromosome- and chromatid-type aberrations. Investigators at two laboratories undertook FISH analyses, each scoring 4000 metaphases per subject. When the data from each laboratory were examined separately, one found slightly more translocations in the miners while the other found fewer. In neither case was the difference significant at the 95% level of confidence. Combining the data likewise showed no significant excess of damage in the miners. This applied to simple one- and two-way translocations and to cells with complex exchanges. There was no correlation between levels of translocations and total lifetime doses from occupational and/or background irradiation. A borderline significant excess of rogue cells was found in the miners. This may be a chance observation, as these rare, highly abnormal cells are considered to be unrelated to radiation exposure and are probably due to a virus. The overall conclusion is that the frequency of chromosomal damage in the miners did not exceed that in the controls. Therefore, the result of the earlier study was not confirmed.

Chromosome Aberrations↗

PubMed2001

[Effect of forest patch size and isolation on reproductive success of Great Tit in fragmented secondary-forests].

The effect of forest patch size and isolation on reproductive success of Great Tit (Parus major) in fragmented secondary-forests was studied in Zuojia Natural Protection Area of Jilin Province by collecting data on breeding success, clutch-size, laying date, egg weight, brood-size and number fledged of Great Tit in 18 large (20-30 hm2), medium (10-20 hm2), and small (6-10 hm2) patches, and two extensive forests (> 100 hm2). All the forest patches were GPS tested. The results showed that isolation had no effect on reproductive success of Great Tit, and Great Tit laid 7.2 days earlier in extensive forests than in all forest patches. Clutches in extensive forests and large forest patches were slightly larger than those in medium and small forest patches, and eggs were heavier in extensive forests than in the patches. Patch size had no effect on brood-size and number fledged. The lost rate of the nests was high in medium and small forest patches because of intense competition.

Animals↗

PubMed2000

Up-regulation of Na,K-ATPase beta 1 transcription by hyperoxia is mediated by SP1/SP3 binding.

The sodium pump, Na,K-ATPase, is an important protein for maintaining intracellular ion concentration, cellular volume, and ion transport and is regulated both transcriptionally and post-transcriptionally. We previously demonstrated that hyperoxia increased Na,K-ATPase beta(1) gene expression in Madin-Darby canine kidney (MDCK) cells. In this study, we identify a DNA element necessary for up-regulation of the Na,K-ATPase beta(1) transcription by hyperoxia and evaluate the nuclear proteins responsible for this up-regulation. Transient transfection experiments in MDCK cells using sequential 5'-deletions of the rat Na,K-ATPase beta(1) promoter-luciferase fusion gene demonstrated promoter activation by hyperoxia between -102 and +151. The hyperoxia response was localized to a 7-base pair region between -62 and -55, which contained a GC-rich region consistent with a consensus sequence for the SP1 family, that was sufficient for up-regulation by hyperoxia. This GC element exhibited both basal and hyperoxia-induced promoter activity and bound both transcription factors SP1 and SP3 in electrophoretic mobility shift assays. In addition, electrophoretic mobility shift assays demonstrated increased binding of SP1/SP3 in cells exposed to hyperoxia while mutation of this element eliminated protein binding. Other GC sites within the proximal promoter also demonstrated up-regulation of transcription by hyperoxia, however, the site at -55 had higher affinity for SP proteins.

Animals↗

PubMed2000

Influence of cellular ganglioside depletion on tumor formation.

BACKGROUND: Gangliosides are immunosuppressive cell surface molecules that are often present in high concentrations in and shed actively by tumor cells. These molecules inhibit the antitumor immune response that is implicated in tumor rejection. We therefore determined the ability of tumor cells pharmacologically depleted of gangliosides to form tumors in mice. METHODS: We tested a ganglioside-rich subline of B16 murine melanoma, MEB4, and MEB4 cells that had been depleted of endogenous gangliosides by incubation with 0.5 microM 1-phenyl-2-hexadecanoylamino-3-pyrrolidino-1-propanol, a specific inhibitor of the enzyme glucosylceramide synthase. Tumor formation was assessed twice a week for 10 weeks after the intradermal injection of tumor cells, and metastatic potential was assessed 4 weeks after tail vein injection of tumor cells. All P values are from two-sided tests. RESULTS: Reduction of the ganglioside content of MEB4 cells, which was not cytotoxic to cells and did not inhibit cell proliferation in vitro, markedly reduced their ability to form tumors. Only 40% of the mice given an intradermal injection of 10(5) ganglioside-depleted MEB4 cells developed tumors compared with 100% of the mice given an injection of 10(5) control MEB4 cells (P<.001). Ganglioside depletion also reduced metastasis: A mean of five pulmonary metastases was detected per mouse given an injection of 2 x 10(5) ganglioside-depleted MEB4 cells compared with a mean of 25 per mouse given an injection of 2 x 10(5) control MEB4 cells. CONCLUSION: Tumor cells with a pharmacologically decreased concentration of gangliosides produce fewer tumors in mice than do untreated cells, suggesting that pharmacologic depletion of gangliosides should be explored further as a therapeutic approach to cancer.

Animals↗

PubMed2000

A novel alpha-helix-liked metallohelicate series and their structural adjustments for the isomorphous substitution.

A novel metallohelical motif is well designed and synthesized by mimicking the alpha-helical fold structure of protein. The 1D helical structures of [Cd(CH2(COO)2)(SC(NH2)2)2]n (I) and [Zn(CH2(COO)2) (SC(NH2)2)2]n (II) are primarily induced and stabilized by the multiple long-range intrahelix hydrogen bonds. Malonate dianion acts as a bidentate ligand coordinated with metal ions to form the backbone of the helix, and thiourea molecules that bend into the helical turn are involved in the intrahelix hydrogen-bond system. The metal ion occupations in the helix of I and II can be freely substituted by simply controlling the initial ratio of those two metal ions. Single crystals of three mixed metal ion complexes of [Cd0.77Zn0.23(CH2(COO)2)(SC(NH2)2)2]n (III), [Cd0.50Zn0.50(CH2(COO)2)(SC(NH2)2)2]n (IV), and [Cd0.21Zn0.79(CH2(COO)2)(SC(NH2)2)2]n (V) were synthesized from systems with an initial Cd/Zn mole ratio of 1:1 for III, 1:2 for IV, and 1:8 for V. They are isomorphous as confirmed by X-ray characterization. When the metal ion is substituted, the multiple intrahelix hydrogen interaction motifs of the coordination polymer structure are self-adjusted to sustain their 1D helical motifs.

Journal Article↗

PubMed2000

Enteropathogenic Escherichia coli (EPEC) attachment to epithelial cells: exploiting the host cell cytoskeleton from the outside.

Enteropathogenic Escherichia coli (EPEC), a leading cause of human infantile diarrhoea, is the prototype for a family of intestinal bacterial pathogens that induce attaching and effacing (A/E) lesions on host cells. A/E lesions are characterized by localized effacement of the brush border of enterocytes, intimate bacterial attachment and pedestal formation beneath the adherent bacteria. As a result of some recent breakthrough discoveries, EPEC has now emerged as a fascinating paradigm for the study of host-pathogen interactions and cytoskeletal rearrangements that occur at the host cell membrane. EPEC uses a type III secretion machinery to attach to epithelial cells, translocating its own receptor for intimate attachment, Tir, into the host cell, which then binds to intimin on the bacterial surface. Studies of EPEC-induced cytoskeletal rearrangements have begun to provide clues as to the mechanisms used by this pathogen to subvert the host cell cytoskeleton and signalling pathways. These findings have unravelled new ways by which pathogenic bacteria exploit host processes from the cell surface and have shed new light on how EPEC might cause diarrhoea.

Adhesins, Bacterial↗

PubMed2000

A comparative study on potential cytogenetic fingerprints for radiation LET in human lymphocytes.

PURPOSE: To carry out a comparative study on potential cytogenetic fingerprints for radiation LET in human metaphase lymphocytes. MATERIALS AND METHOD: Human lymphocytes were irradiated in vitro with 3.0 Gy 60Co gamma-rays, 0.9 Gy 3H beta-rays or 0.2 Gy 2.7 Mev neutrons. Detailed chromosome aberrations were analysed by combined FISH with pan-telomere staining and specific whole-chromosome painting (1, 2 and 4). Total chromosome translocations and insertions were also analysed by multicolour whole-chromosome painting (chromosomes 1, 2 and 4 orange, chromosomes 3, 5 and 6 green). RESULTS: Among the six proposed radiation cytogenetic fingerprints, the ratio of total simple translocations to insertions (I-ratio), showed the largest difference between low-LET 60Co gamma-ray and high-LET neutron radiation. The ratios of complete exchanges to incomplete rejoinings [S(I)-ratio] and dicentrics to interstitial deletions (H-ratio), showed a similar significant difference between low- and high-LET radiation. The ratios of centric rings to interstitial deletion (G-ratio) showed a trend of LET-related difference, but the difference was not significant in this data set. The ratios of dicentrics to centric rings (F-ratio) and apparent complete exchanges to hidden complete exchanges [S(II)-ratio], showed no difference between low- and high-LET radiation. In the 1426 radiation-induced chromosome aberrations observed after 52 h culture, evidence for sister-chromatid fusion but not telomere addition was found. CONCLUSION: Pan-telomere staining plus specific whole chromosome painting allows simultaneous and objective detection of complete or incomplete chromosome exchanges and interstitial or terminal deletions in human peripheral lymphocytes. Of the six proposed cytogenetic ratios, the I-ratio is the most effective cytogenetic fingerprint for distinguishing low-LET from high-LET radiation in human metaphase human lymphocytes.

Beta Particles↗

PubMed2000

The interaction of lead exposure and arylsulfatase A genotype affects sulfatide catabolism in human fibroblasts.

Lead exposure causes cognitive and behavioral deficits in some affected children. We propose that a contributing mechanism for the neurological damage is that lead induces critically low levels of arylsulfatase A (ASA) at sensitive stages of nervous system development. It is hypothesized that the combined effects of a single nucleotide polymorphism (SNP) in human ASA which results in reduced levels of the enzyme, and lead concentrations which decrease ASA activity culminate in cellular enzymic activity that is below a critical threshold required for the maintenance of normal nervous system function. Human fibroblasts grown in the presence of 20 microM lead acetate exhibit a more than 60% decrease of cellular ASA enzyme protein. Lead treatment of cells from individuals with the SNP(s) of pseudodeficient ASA, but not those from subjects with the normal gene, results in a significant decrease in ability of the cells to desulfate sulfatide, the substrate of ASA. The decrease in the degree of sulfatide catabolism is consistent with possible enhanced lead-induced neurobehavioral effects in individuals homozygous for the pseudodeficiency polymorphism(s) of ASA.

Cells, Cultured↗

PubMed2000

[Effect of jiaweiyiguanjian decoction on hypothalamus-pituitary-thyroid gland (HPT) in rat model with Yin-deficiency of liver and kidney induced by slow irritation].

OBJECTIVE: To explore the mechanism of Jiaweiyiguanjian decoction in nourishing the liver and kidney. METHOD: A rat model with Yin-deficiency of liver and kidney was made by way of slow irritation. Thyrotropin-releasing-hormone(TRH) of thyron and blood, TSH of pituitary and blood, thyroxine, 3,5,3',5-tetraiodothyronine(FT4) and 3,3',5-traiodothyronine(FT4), 3,3',5-traiodothyronine(rT3) were used as indexes to study the effect of the decoction on HPT. RESULT: The TRH Secretion from hypothalamus increased (P < 0.05), TSH of pituitary and blood reduced(P < 0.05), FT3 and FT4 of blood decreased at the same time and rT3 of blood, increased. The indexes of the treatment group were found almost the same as the those of the normal control group. CONCLUSION: Jiaweiyiguanjian Decoction could adjust HPT.

Animals↗

PubMed2000

[cDNA cloning and sequence analysis of the seventh segment of maize rough dwarf virus genome].

The double strand RNA of maize rough dwarf virus (MRDV) was prepared from the maize samples showing symptoms which was from the Luanchen county of Heibei province of China. The primers were designed according to the known sequence of MRDV, the cDNA sequence of the seventh segment of MRDV was obtained by RT-PCR, the S7 sequence was analyzed by computer after sequencing. The results showed: the full length of the S7 cDNA is 1936 bp and equal to that of the S7 cDNA from abroad, the two open reading frame(ORF1 and ORF2) contained in the S7 segment are also unchanged. In comparison with the S7 segment from Italy, the homology of S7 nucleotide is 87.7% and the homology of ORF1 amino acid sequence is 91.6%. However, the MRDV S7 segment and the rice black strike dwarf virus S8 segment showed 95.5% nucleotide identities and 93.5% ORF1 amino acid identities.

Amino Acid Sequence↗

PubMed1999

Influence of 5-bromodeoxycytosine substitution on triplex DNA stability and conformation.

Three triple-helical hairpin DNAs with substitution of 5-bromocytosine for cytosine in different strands have been investigated by molecular mechanics and Raman spectroscopy. The stability of the three substituted triplexes were compared with the corresponding unsubstituted triplex DNA by the molecular mechanics method. Base stacking interactions and strand--strand interactions of each triplex were analyzed in detail. Sugar conformations in these triplexes have been determined by both vibrational spectroscopy and molecular dynamics simulation. The hairpin triplexes with substitution occurring in strand I or both in strands I and III have the main sugar conformation of C3'-endo, while the triplex with substitution occurring in strand III is the combination of C3'-endo and C2'-endo sugar conformation. Theoretical results are basically in agreement with experiments.

Base Sequence↗

PubMed1999

VirE1 is a specific molecular chaperone for the exported single-stranded-DNA-binding protein VirE2 in Agrobacterium.

Agrobacterium tumefaciens induces tumours on plants by transferring a nucleoprotein complex, the T-complex, from the bacterium to the plant cell. The T-complex consists of a single-stranded DNA (ssDNA) segment, the T-DNA, and VirD2, an endonuclease covalently attached to the 5' end of the T-DNA. A type IV secretion system encoded by the virB operon and virD4 is required for the entry of the T-complex and VirE2, a ssDNA-binding protein, into plant cells. The VirE1 protein is specifically required for the export of the VirE2 protein, as demonstrated by extracellular complementation and tumour formation. In this report, using a yeast two-hybrid system, we demonstrated that the VirE1 and VirE2 proteins interact and confirmed this interaction by in vitro binding assays. Although VirE2 is a ssDNA-binding protein, addition of ssDNA into the binding buffer did not interfere with the interaction of VirE1 and VirE2. VirE2 also interacts with itself, but the interaction between VirE1 and VirE2 is stronger than the VirE2 self-interaction, as measured in a lacZ reporter gene assay. In addition, the interaction of VirE2 with itself is inhibited by VirE1, indicating that VirE2 binds VirE1 preferentially. Analysis of various virE2 deletions indicated that the VirE1 interaction domain of VirE2 overlaps the VirE2 self-interaction domain. Incubation of extracts from Escherichia coli overexpressing His-VirE1 with the extracts of E. coli overexpressing His-VirE2 increased the yield of His-VirE2 in the soluble fraction. In a similar purified protein solubility assay, His-VirE1 increased the amount of His-VirE2 partitioning into the soluble fraction. In Agrobacterium, VirE2 was undetectable in the soluble protein fraction unless VirE1 was co-expressed. When urea was added to solubilize any large protein aggregates, a low level of VirE2 was detected. These results indicate that VirE1 prevents VirE2 from aggregating, enhances the stability of VirE2 and, perhaps, maintains VirE2 in an export-competent state. Analysis of the deduced amino acid sequence of the VirE1 protein revealed that the VirE1 protein shares a number of properties with molecular chaperones that are involved in the transport of specific proteins into animal and plant cells using type III secretion systems. We suggest that VirE1 functions as a specific molecular chaperone for VirE2, the first such chaperone linked to the presumed type IV secretion system.

Bacterial Proteins↗

PubMed1999

Combined FISH with pan-telomeric PNA and whole chromosome-specific DNA probes to detect complete and incomplete chromosomal exchanges in human lymphocytes.

PURPOSE: To combine FISH with pan-telomeric peptide nucleic acid (PNA) and whole chromosome-specific DNA probes to detect complete and incomplete chromosome exchanges in human lymphocytes. MATERIALS AND METHODS: Human lymphocytes were irradiated in vitro with 0.9 Gy low dose-rate (0.019 Gy/h) tritium beta-rays. Metaphase spreads were treated with RNase, fixed in 1:3 acetic acid:methanol, and then further treated with KCl, proteinase K and fixed in 4% paraformaldehyde. Slides were denatured, hybridized for 1.5 h with an FITC-labelled telomeric PNA probe, and rehybridized overnight with a spectrum-orange whole-chromosome probe specific for chromosomes 1, 2 and 4. Hybridized spreads were washed with 70% formamide/20 x SSC and counterstained with DAPI. RESULTS: All three pairs of labelled chromosomes together with 92 telomeres were readily visible after hybridization. The whole chromosomes 1, 2 and 4 were painted orange, and all telomeres were stained green. Unpainted chromosomes were counterstained blue. In the observed 680 chromosome aberrations induced by tritium beta-rays in human lymphocytes after 52 h of culture, no evidence of telomere addition was detected. Incomplete and hidden complete exchanges and terminal deletions were definitively discriminated. CONCLUSION: The simultaneous detection of telomeres and specific whole chromosomes allows for the first time accurate analysis of complete and incomplete chromosome exchanges involving painted chromosomes in human lymphocytes.

Cells, Cultured↗

PubMed1999

Background ionizing radiation plays a minor role in the production of chromosome translocations in a control population.

PURPOSE: To obtain a relationship between background chromosome translocation frequency and age with translocation frequency measured to a high statistical precision, and to identify the role of background ionizing radiation in the production of chromosome translocations in a control population. MATERIALS AND METHODS: Lymphocytes from 35 healthy control individuals (15 females and 20 males) were scored, using fluorescence in situ hybridization, for the presence of chromosomal translocations. Translocation frequencies were measured to a high statistical precision (s.d. 25% or less for each individual). These control subjects were of varying ages, ranging from 0 (cord blood) to 98 years. RESULTS: In a total of 521,492 metaphases (203,754 genome equivalent cells) scored, an average of 5,822 genome equivalent cells per individual, 764 translocations were observed in the 35 individuals. The translocation frequencies ranged from 0 (for cord blood) to 0.0167 (for a 98 year old) translocations per cell. The average age and translocation frequency was 50 years and 0.004 translocations per cell, respectively. The best fit of the relationship between translocations and age was: Y=7x10(-4)+6.9x10(-6)A+1.35x10(-6)A2, which does not obey the linear relationship expected from chronic background radiation alone. The curvilinear relationship observed clearly shows that other endogenous and exogenous clastogens or clastogenic events, in addition to radiation, serve to generate chromosome translocations in control populations. CONCLUSION: The background translocation frequency in control individuals follows a curvilinear relationship with age. No significant variation was observed between individuals of the same age. Clastogenic processes of normal aging and physiological factors in additional to ionizing radiation play a major role in the production of chromosome translocations in a control population. Background radiation, however, appears to play a minor role in chromosome translocation production in control individuals living near sea level.

Adult↗

PubMed1999

Exposure temperature, but not donor age, is a confounding factor for in vitro translocation production by chronic irradiation.

PURPOSE: To assess the effects of incubation temperature during irradiation, and of donor age, on the in vitro induction of chromosomal translocations in human lymphocytes. MATERIAL AND METHODS: Lymphocytes from six human male donors were scored, using fluorescence in situ hybridization, for the presence of chromosomal translocations involving chromosomes 1 to 6 after in vitro, chronic exposure (delivered continuously over 48 h at 37 degrees C or at 20 degrees C) to tritium beta-rays or 60Co gamma-rays. RESULTS: No age-related difference in the alpha coefficients of the fitted induction curves was observed for gamma-ray-exposed lymphocytes obtained from four donors whose ages ranged from 24 to 79 years, or for tritium beta-ray-exposed lymphocytes from two donors aged 36 and 62 years. Duplicate samples from one donor, irradiated concurrently at 20 degrees C or 37 degrees C, gave significantly different alpha coefficients: 0.128+/-0.008 and 0.053+/-0.004, respectively (p<0.0001). The S-ratio (the ratio of induced complete to incomplete translocations) was found to be independent of radiation dose, donor age and exposure temperature. CONCLUSIONS: For biodosimetry in chronic irradiation situations, the use of alpha coefficients derived from the dose-response curves of cells chronically irradiated in vitro at body temperature is recommended. With respect to induction rates, donor age does not appear to be a confounding factor. The S-ratio is independent of radiation doses, exposure temperatures, or donor ages.

Adult↗