Genetic ecotoxicology I: DNA integrity and reproduction in mosquitofish exposed in situ to radionuclides

Female mosquitofish (Gambusia affinis) were collected from two sites located on the US Department of Energy's Oak Ridge Reservation that are contaminated with 137Cs, 90Sr, other radionuclides and chemical genotoxicants. Fish from non- radionuclide contaminated environments located off the reservation were also collected. DNA, extracted from liver tissue and blood cells, was examined by gel electrophoresis for structural damage in the form of strand breakage. In general, the level of DNA strand breaks was elevated in fish from radionuclide-contaminated sites with observed differences in the number and type of strand breaks between liver tissue and blood cells. The number of malformed embryos was higher in fish at the contaminated sites, and varied with season. Fecundity was negatively correlated with the level of double strand breaks in the DNA of fish from one contaminated site. Females with broods that included malformed embryos had more DNA strand breakage than those that did not; and furthermore, a threshold effect was observed between the occurrence of malformed embryos and the presence of double strand breaks in the DNA of the mother. These findings have implications for both ecological risk assessment and evolutionary ecology     

Experiences with risk estimates for carriers of chromosomal reciprocal translocations

The risk estimates for individual carriers of ten different familial reciprocal translocations detected among 500 couples with reproductive failures are presented. These were established by application of the empirical data analysed by Stengel-Rutkowski et al. (1988) and the guidelines given in Stene & Stengel-Rutkowski (1988). Different risks were estimated for unbalanced offspring at birth or at second trimester prenatal diagnosis for abortions, or stillbirths/early deaths. These risk estimates varied considerably from translocation to translocation. Carriers of five translocations had risks for offspring with single-segment imbalances. The birth risk figures ranged from 0.1% to 13.8%. Carriers of five other translocations had risks for double-segment imbalances with birth risks ranging from 0% to 3.2%. The estimated risk figures were independent of the method of ascertainment. Among the parents of the index cases we found nine maternal carriers and only one paternal carrier. This presentation illustrates the need for individual risk counselling of each carrier with reciprocal translocation regarding further family planning.     

Suggestive linkage to chromosome 19 in a large Cuban family with late‐onset Parkinson's disease

The identification of disease genes using family‐based approaches has provided important insights into the pathogenesis of Parkinson's disease (PD) demonstrating the importance of genetic studies on monogenic forms of the disease. We studied a large Cuban family with typical, late‐onset PD and probable autosomal dominant inheritance. Mean age at onset was 61.2 years (±12.53, 45–76). Other phenotypes such as essential tremor and atypical parkinsonism were observed in this family. We carried out a genome‐wide scan and linkage analyses. The genetic data were analyzed using a conservative model in which only patients with clinically definite or likely PD were considered affected, other phenotypes were regarded as “unknown.” Multipoint analyses yielded a maximum LOD of 2.26 between markers D19S221 and D19S840. Haplotype analysis showed a region on chromosome 19 shared by six of seven PD patients. The essential tremor phenotype and the atypical parkinsonism do not segregate with this haplotype, suggesting a different etiology. Our findings suggest the presence of a novel locus for PD on chromosome 19p13.3–q12. We propose that an oligogenic model with moderate contribution of two or three genes rather than a “pure” monogenic model might explain better the wide range in age at onset, the reduced penetrance and the phenotypical variability observed in PD families. © 2003 Movement Disorder Society     

Why do long-distance runners not have more bone? A vital biomechanical explanation and an estrogen effect

Inanimate structures cannot detect and repair their fatigue damage or microdamage, so to minimize it they need more structural material and strength. Living bone handles this matter differently. Bone modeling drifts adapt bone architecture and strength to the loads on bones in ways that tend to keep strains from exceeding a “modeling threshold” range. Strains (or equivalent features) above that threshold switch mechanically controlled modeling ON. Where strains stay below that threshold, this modeling goes OFF. Repeatedly loading-deloading a bone causes microdamage in it, and basic multicellular unit (BMU)-based bone remodeling normally repairs it. Where strains stay below an operational “microdamage threshold,” remodeling can repair whatever microdamage happens for as long as it happens. Strains above that threshold can cause too much microdamage to repair completely and lead to fatigue fractures of trabeculae or whole bones. The modeling threshold normally lies comforably below the microdamage threshold. Since modeling normally adjusts bone architecture to keep strains from exceeding the modeling threshold, this keeps strains below the microdamage threshold, too, and voluntary activities do not cause more microdamage than remodeling can repair. Therefore, long-distance runners do not need more bone mass and strength than nonrunners of comparable age, sex, and body size.     

Simultaneous genetic analysis of longitudinal means and covariance structure in the simplex model using twin data

A longitudinal model based on the simplex model is presented to analyze simultaneously means and covariance structure using univariate longitudinal twin data. The objective of the model is to decompose the mean trend into components which can be attributed to those genetic and environmental factors which give rise to phenotypic individual differences and a component of unknown constitution which does not involve individual differences. Illustrations are given using simulated data and repeatedly measured weight obtained in a sample of 82 female twin pairs on six occasions.     

Ninety-one Cases of Breast Cancer and Chronic Lymphoproliferative Neoplasm: A Retrospective Review of a Population at High Risk for Multiple Malignancies

Abstract: We performed a retrospective analysis of clinical course of 91 patients who developed both breast cancer and a chronic lymphoproliferative neoplasm and were seen at the M. D. Anderson Cancer Center between January 1, 1970 and December 30, 1991. The sample included 24 individuals who developed lymphoproliferative neoplasm first (Group A), 22 individuals with concurrent diagnosis of both malignancies (Group B), and 45 individuals who developed breast cancer first (Group C). The median time to diagnosis of secondary breast cancer and lymphoproliferative neoplasm was 66 months (range, 7–459) and 65 months (range, 0–334), respectively. A higher proportion of Group B lymphomas were low-grade (77% vs. 47% [Group A] vs. 37% [Group C] p = 0.009). Prior occurrence of either one of these malignancies did not affect the disease-specific survival from the second malignancy. However, continuing mortality from the first malignancy appeared to contribute to a poor overall survival following second malignancy. Group A included 8 patients who developed breast cancer following radiation therapy for Hodgkin's disease after a mean interval of 18 (± 4.3) years. Three of these individuals had coexisting ductal and lobular histology (vs. none of the individuals in Groups B and C, p = 0.02). Another interesting finding was the high incidence of multiple additional malignancies in this patient population. A total of 29 additional neoplasms occurred in 21 (23%) of the 91 study subjects. These malignancies involved a wide variety of organ sites and could not be attributed to the therapy for either the breast cancer or the lymphoma in most cases. The data suggest that individuals who develop both breast cancer and a lymphoproliferative neoplasm are at a high risk for multiple malignancies. Close surveillance of such individuals for additional malignancies and further studies to understand the molecular basis of this predisposition are warranted.?     

Predictive testing for Huntington's disease: I. Predictors of uptake in South Wales

In Wales, predictive testing for Huntington's disease (HD) has not been offered proactively to families and uptake of testing is low in comparison to other centres. Little is known of those not requesting testing, particularly those not in direct contact with the genetics service. This study examined differences between a cohort of 22 test applicants and a random group of 32 'non-requesters', drawn from the South Wales HD register. Respondents were interviewed by means of a semi-structured schedule in their own homes. The study groups differed significantly on a number of variables including: knowledge of the availability of testing; perceived attitudes of family members and significant others to testing; length of knowledge and perceived stressfulness of being at risk; and perceived ability to cope with an unfavourable result. Overall, knowledge of testing procedures was poor and at-risk individuals' understanding of genetic terminology was at odds with scientific distinctions. Discussion focuses on the organisational and psychological factors associated with lack of knowledge of the availability of testing and the interpretation of reported coping capacities.     

Radiofrequency Current Ablation of Porcine Right Atrium: Increased Lesion Size with Bipolar Two Catheter Technique Compared to Unipolar Application In Vitro and In Vivo

Interruption of atrial flutter and fibrillation by RF catheter ablation may be favored by large, elongated lesions. We administered RF current in unipolar and bipolar mode in porcine right atrium. Bipolar ablation was performed between the tip electrodes of two serially coupled catheters. With 4-mm tip electrodes in vitro, lesion length increased from a mean (SD) of 7.9 (1.2) mm at 3 mm-interelectrode distance (IED) to 13.3 (3.3) mm at 9-mm IED, but decreased at 12-mm IED due to nonconfluent lesions (P < 0.0001), With 4 mm distal electrodes and 8 mm IED, bipolar lesions were 65% longer than corresponding unipolar ablations. Switching to bipolar mode increased the lesion length more than increasing electrode tip length to 6 mm in unipolar mode. Power and temperature controlled ablation created equally sized lesions. Twelve anesthetized pigs were randomized to unipolar or two catheter bipolar temperature controlled ablation of the right atrial free wall. Bipolar ablation created confluent lesions with endocardial length × width of 13.5 (5.8) × 7.3 (3.7) mm, unipolar ablation 6.4 (2.8) × 4.6 (1.4) mm (P < 0.001 when comparing length and P = 0.013 for lesion width). The atrial lesions in both groups were transmural and extended into hilar lung lesions with maximal depth of 3.0 (1.1) and 2.6 (1.0) mm, respectively (P = 0.44). Five bipolarly and four unipolarly ablated pigs developed right diaphragmal paresis. We conclude that bipolar ablation may be preferable in situations where large, elongated lesions are favorable. The two catheter technique is feasible in porcine right atrium. Both bipolar and unipolar ablation of the porcine right atrial free wall may frequently be complicated by injury to the phrenic nerve and adjacent lung tissue.     

巨噬细胞炎性蛋白4的突变和原核表达

目的 探索如何抑制嗜酸细胞的趋化作用，选择β-趋化因子巨噬细胞炎性蛋白4（MIP4）的突变性（Met-MIP4)作为趋化因子受体3的拮抗剂，将Met-MIP4基因在原核细胞中进行表达。方法 设计MIP4基因的PCR引物并进行氨基酸突变，将MIP4N末端的丙氨酸突变为蛋氨酸，以正常人肺酸突变，将MIP4N末端的丙氨酸突变为蛋氨酸。以正常人肺cDNA文库为模板，PCR方法获取Met-MIP4基因，克隆入载体pUC19，测序验证序列已得到突变，将正确的基因插入到GST融合表达载体pGEX-4T中，以IPTG诱导表达。结果 PCR产物为220bp左右的片段，连接入pUC19质粒后测序验证获得正确突变，构建的pGEX-4T融合表达载体在大肠杆菌中表达，经SDS-PAGE凝胶电泳显示有大小约34kU的新生融合蛋白表达。结论 成功突变并克隆了β-趋化因子MIP4基因，SDS-PAGE表明，与GST融合的Met-MIP4突变体已得到表达，为进一步研究其生物学活性奠定了基础。     

Comparative studies on cytotoxic and genotoxic effects of two organic mercury compounds in lymphocytes and gastric mucosa cells of sprague-dawley rats

Human lymphocytes (HL) as well as lymphocytes (RL), hepatocytes (RH), and gastric mucosa cells (GM) of Sprague-Dawley rats were treated in vitro for 1 h with methylmercury chloride (MMC, 0.5–4 μg/ml) and dimethylmercury (DMM, 5–40 μg/ml). The cytotoxicity of the two organic mercury compounds was assessed by dye exclusion, and the extent of induced DNA fragmentation was measured with a single-cell microgel electrophoresis assay. Both MMC and DMM induced DNA damage and cytotoxicity in a dose-related manner in HL, RL, and GM. MMC was more effective in causing a significant increase in median DNA migration than DMM at doses yielding approximately the same degree of cytotoxicity. In rat hepatocytes the MMC-induced DNA damage was, however, lower than in the other cells. An analysis of repair kinetics following exposure to 2 μg/ml MMC was carried out in human lymphocytes obtained from an adult male donor. The bulk of DNA repair occurred 90 min after in vitro exposure, and it was about complete by 120 min following cessation of exposure. Finally, in order to have a basis for extrapolating to the human situation, in vivo studies were performed with Sprague-Dawley rats, also assessing the DNA damage and cytotoxicity in the lymphocytes and gastric mucosa cells. These in vivo results after oral exposure may be directly compared to the in vitro data obtained in the same cells. © 1993 Wiley-Liss, Inc.