Evidence of linkage disequilibrium in the Spanish polycystic kidney disease I population.

Forty-one Spanish families with polycystic kidney disease 1 (PKD1) were studied for evidence of linkage disequilibrium between the disease locus and six closely linked markers. Four of these loci--three highly polymorphic microsatellites (SM6, CW3, and CW2) and an RFLP marker (BLu24)--are described for the first time in this report. Overall the results reveal many different haplotypes on the disease-carrying chromosome, suggesting a variety of independent PKD1 mutations. However, linkage disequilibrium was found between BLu24 and PKD1, and this was corroborated by haplotype analysis including the microsatellite polymorphisms. From this analysis a group of closely related haplotypes, consisting of four markers, was found on 40% of PKD1 chromosomes, although markers flanking this homogeneous region showed greater variability. This study has highlighted an interesting subpopulation of Spanish PKD1 chromosomes, many of which have a common origin, that may be useful for localizing the PKD1 locus more precisely.     

Evaluation of a cosmid contig physical map of human chromosome 16.

A cosmid contig physical map of human chromosome 16 has been developed by repetitive sequence finger-printing of approximately 4000 cosmid clones obtained from a chromosome 16-specific cosmid library. The arrangement of clones in contigs is determined by (1) estimating cosmid length and determining the likelihoods for all possible pairwise clone overlaps, using the fingerprint data, and (2) using an optimization technique to fit contig maps to these estimates. Two important questions concerning this contig map are how much of chromosome 16 is covered and how accurate are the assembled contigs. Both questions can be addressed by hybridization of single-copy sequence probes to gridded arrays of the cosmids. All of the fingerprinted clones have been arrayed on nylon membranes so that any region of interest can be identified by hybridization. The hybridization experiments indicate that approximately 84% of the euchromatic arms of chromosome 16 are covered by contigs and singleton cosmids. Both grid hybridization (26 contigs) and pulsed-field gel electrophoresis experiments (11 contigs) confirmed the assembled contigs, indicating that false positive overlaps occur infrequently in the present map. Furthermore, regional localization of 93 contigs and singleton cosmids to a somatic cell hybrid mapping panel indicates that there is no bias in the coverage of the euchromatic arms.     

Increased alertness behavior in Australian fur seals (Arctocephalus pusillus doriferus) at a high vessel traffic haul-out site

Vessel impacts on marine mammals are of growing concern, and marine mammals in urbanized marine environments are at particular risk of exposure. Port Phillip Bay (Victoria, Australia) is one such environment, in which Australian fur seals (AUFS; Arctocephalus pusillus doriferus) haul-out to rest, yet little is known about the impacts of vessels on resting seals. We used remote camera traps to investigate the influence of vessel traffic on AUFS behavior at a nonbreeding haul-out site. Environmental, temporal, and vessel-related variables were all associated with changes in AUFS alertness at this site. All vessel types elicited increased alertness above base-line levels (25%), with recreational and commercial motorized vessels associated with a 5.7%–10.8% increase in alertness. Unidentified vessels, the government vessel, and kayaks were associated with significantly increased alertness of 21.7%, 46.4%, and 60.7%, respectively, though accounted for only 6.2% of vessel observations. Vessels breaching current approach regulations (<5 m) showed a 32% increase in alertness, significantly higher than nonbreach approaches. Partial and complete flushing of the platform was rare, occurring in 1.0% of images analyzed. These results suggest that vessels do elicit a response from AUFS at this haul-out site, and that further monitoring of vessel activity and compliance is required.     

Fecal steroid analysis of ovarian cycles in free-ranging baboons

This paper reports field and laboratory tests of serial sampling, solid phase extraction, and microradioimmunoassay methods for the collection, preservation, and analysis of fecal steroids. The field study was conducted in a troop of 87 yellow baboons (Papio cynocephalus) in the Tana River Primate Reserve, Kenya. Serial samples of four focal females and opportunistic sampling of 18 additional females over 22 days of sampling yielded a total of 62 samples, X = 3.1 ± 0.4/day, demonstrating the feasibility of regular field collection and extraction. Estradiol and progesterone concentrations in the field-extracted samples exhibited high recovery and statistically significant correlations (P < 0.05) with concentrations in the lab-extracted samples, suggesting that solid phase extraction could provide a useful alternative to freezing in sites where electricity or liquid nitrogen is not available. Tests of microradioimmunoassays demonstrated that these assays were sensitive, accurate, and precise when applied to the assay of fecal extracts, providing estimates of ovarian steroids that varied significantly with reproductive state. The demonstration that testosterone could be accurately and reliably assayed in fecal extracts suggests that these techniques also could be applied to the study of male reproductive function. Parallels between fecal profiles of cycling and pregnant baboons with patterns reported for serum steroids in baboons suggest that fecal steroids might be useful in distinguishing amenorrhea from early pregnancy in free-ranging baboons as well as in species lacking external indices of reproductive state. © 1995 Wiley-Liss, Inc.     

Analysis of hybrids between an H-2+, TL− lymphoma and an H-2+, TL+ lymphoma and its H-2−, TL− variant subline

Hybrids between an H-2⁺, TL⁺ lymphoma and an H-2⁺, TL^– lymphoma were studied for their expression of H-2 and TL antigens. The H-2 antigens of both parents were expressed, the TL specificity of the TL⁺ parent was retained, and the TL specificity characteristic of the mouse strain from which the TL^– lymphoma was derived was not expressed. There was no evidence that the genome of either parent altered the expression of the TL antigens coded for by the genome of the opposite parent. Hybrids between the H-2⁺, TL^– lymphoma and an H-2^–, TL^– variant line derived from the H-2⁺, TL⁺ cell line expressed both theK- andD-regioncoded H-2 antigens and the TL specificity characteristic of the parental cell line from which the variant cell was derived. This result is consistent with the defect in the variant cell's being the result of a mutation affecting a gene coding for a positive element necessary for expression of both TL and the serologically detectable H-2 antigens on the cell surface.     

Determination of subject-specific model parameters for visco-elastic elements

The determination of subject-specific model parameter values is necessary in order for a computer simulation model of human motion to be evaluated quantitatively. This study used an optimisation procedure along with a kinematically driven simulation model of the contact phase in running jumps to determine the elastic parameters of segmental wobbling masses and the foot-ground interface. Kinetic and kinematic data were obtained on running jumps for height and distance performed by an elite male high jumper. Stiffness and damping coefficients of the visco-elastic elements in the model were varied until the difference between simulation and performance was minimised. Percentage differences of 6% and 9% between the simulated and recorded performances were obtained in the jumps for height and distance, respectively. When the parameters obtained from the jump for height were used in a simulation of the jump for distance (and vice versa), there was poor agreement with the recorded jump. On the other hand, a common set of visco-elastic parameters were obtained using the data from both recorded jumps resulting in a mean difference of only 8% (made up of 7% and 10%) between simulation and performance that was almost as good as the individual matches. Simulations were not overly sensitive to perturbations of the common set of visco-elastic parameters. It is concluded that subject-specific elastic parameters should be calculated from more than a single jump in order to provide a robust set of values that can be used in different simulations.     

Evaluation of a torque-driven model of jumping for height

This study used an optimization procedure to evaluate an 8-segment torque-driven subject-specific computer simulation model of the takeoff phase in running jumps for height. Kinetic and kinematic data were obtained on a running jump performed by an elite male high jumper. Torque generator activation timings were varied to minimize the difference between simulation and performance in terms of kinematic and kinetic variables subject to constraints on the joint angles at takeoff to ensure that joints remained within their anatomical ranges of motion. A percentage difference of 6.6% between simulation and recorded performance was obtained. Maximizing the height reached by the mass center during the flight phase by varying torque generator activation timings resulted in a credible height increase of 90 mm compared with the matching simulation. These two results imply that the model is sufficiently complex and has appropriate strength parameters to give realistic simulations of running jumps for height.     

Nutritional supplement chromium picolinate generates chromosomal aberrations and impedes progeny development in Drosophila melanogaster

Chromium picolinate, [Cr(pic)(3)], is a popular nutritional supplement found in a variety of consumer products. Despite its popularity, safety concerns over its use have arisen. The supplement has been shown to generate clastogenic damage, mitochondrial damage, oxidative damage, and mutagenic effects in cultured cells and oxidative DNA damage and lipid peroxidation in rats. Recently [Cr(pic)(3)] has been demonstrated to generate heritable genetic change and delays in progeny development in Drosophila melanogaster. Based on the damage to chromosomes of cultured cells and of animal models, similar chromosome damage appeared to be a likely source of the mutagenic effects of the supplement in Drosophila. The current three-part study examines the effects of several chromium-containing supplements and their components on hatching and eclosion rates and success of development of first generation progeny of adult Drosophila fed food containing these compounds. It further examines the effects of the compounds on longevity of virgin male and female adults. Finally, the chromosomes in the salivary glands of Drosophila late in the third instar larval stage, which were the progeny of Drosophila whose diets were supplemented with nutritional levels of [Cr(pic)(3)], are shown to contain on average over one chromosomal aberration per two identifiable chromosomal arms. No aberrations were observed in chromosomes of progeny of untreated flies. The results suggest that human consumption of the supplement should be a matter of concern and continued investigation to provide insight into the requirements of chromium-containing supplements to give rise to genotoxic effects.     

Comparative genomic and proteomic analysis of high grade glioma primary cultures and matched tumor in situ

Developing targeted therapies for high grade gliomas (HGG), the most common primary brain tumor in adults, relies largely on glioma cultures. However, it is unclear if HGG tumorigenic signaling pathways are retained under in-vitro conditions. Using array comparative genomic hybridization and immunohistochemical profiling, we contrasted the epidermal and platelet-derived growth factor receptor (EGFR/PDGFR) in-vitro pathway status of twenty-six primary HGG cultures with the pathway status of their original HGG biopsies. Genomic gains or amplifications were lost during culturing while genomic losses were more likely to be retained. Loss of EGFR amplification was further verified immunohistochemically when EGFR over expression was decreased in the majority of cultures. Conversely, PDGFRα and PDGFRβ were more abundantly expressed in primary cultures than in the original tumor (p<0.05). Despite these genomic and proteomic differences, primary HGG cultures retained key aspects of dysregulated tumorigenic signaling. Both in-vivo and in-vitro the presence of EGFR resulted in downstream activation of P70s6K while reduced downstream activation was associated with the presence of PDGFR and the tumor suppressor, PTEN. The preserved pathway dysregulation make this glioma model suitable for further studies of glioma tumorigenesis, however individual culture related differences must be taken into consideration when testing responsiveness to chemotherapeutic agents.