Predicted genetic burden and frequency of phenotype-associated variants in the horse.

Disease-causing variants have been identified for less than 20% of suspected equine genetic diseases. Whole genome sequencing (WGS) allows rapid identification of rare disease causal variants. However, interpreting the clinical variant consequence is confounded by the number of predicted deleterious variants that healthy individuals carry (predicted genetic burden). Estimation of the predicted genetic burden and baseline frequencies of known deleterious or phenotype associated variants within and across the major horse breeds have not been performed. We used WGS of 605 horses across 48 breeds to identify 32,818,945 variants, demonstrate a high predicted genetic burden (median 730 variants/horse, interquartile range: 613-829), show breed differences in predicted genetic burden across 12 target breeds, and estimate the high frequencies of some previously reported disease variants. This large-scale variant catalog for a major and highly athletic domestic animal species will enhance its ability to serve as a model for human phenotypes and improves our ability to discover the bases for important equine phenotypes.

Structural analysis of the mechanism of adenovirus binding to its human cellular receptor, CAR.

Binding of virus particles to specific host cell surface receptors is known to be an obligatory step in infection even though the molecular basis for these interactions is not well characterized. The crystal structure of the adenovirus fiber knob domain in complex with domain I of its human cellular receptor, coxsackie and adenovirus receptor (CAR), is presented here. Surface-exposed loops on knob contact one face of CAR, forming a high-affinity complex. Topology mismatches between interacting surfaces create interfacial solvent-filled cavities and channels that may be targets for antiviral drug therapy. The structure identifies key determinants of binding specificity, which may suggest ways to modify the tropism of adenovirus-based gene therapy vectors.

Studies of activated GPIIb/IIIa receptors on the luminal surface of adherent platelets. Paradoxical loss of luminal receptors when platelets adhere to high density fibrinogen.

The accessibility of activated GPIIb/IIIa receptors on the luminal surface of platelets adherent to damaged blood vessels or atherosclerotic plaques is likely to play a crucial role in subsequent platelet recruitment. To define better the factors involved in this process, we developed a functional assay to assess the presence of activated, luminal GPIIb/IIIa receptors, based on their ability to bind erythrocytes containing a high density of covalently coupled RGD-containing peptides (thromboerythrocytes). Platelets readily adhered to wells coated with purified type I rat skin collagen and the adherent platelets bound a dense lawn of thromboerythrocytes. With fibrinogen-coated wells, platelet adhesion increased as the fibrinogen-coating concentration increased, reaching a plateau at about 11 micrograms/ml. Thromboerythrocyte binding to the platelets adherent to fibrinogen showed a paradoxical response, increasing at fibrinogen coating concentrations up to approximately 4-6 micrograms/ml and then dramatically decreasing at higher fibrinogen-coating concentrations. Scanning electron microscopy demonstrated that the morphology of platelets adherent to collagen was similar to that of platelets adherent to low density fibrinogen, with extensive filopodia formation and ruffling. In contrast, platelets adherent to high density fibrinogen showed a bland, flattened appearance. Immunogold staining of GPIIb/IIIa receptors demonstrated concentration of the receptors on the filopodia, and depletion of receptors on the flattened portion of the platelets. Thus, there is a paradoxical loss of accessible, activated GPIIb/IIIa receptors on the luminal surface of platelets adherent to high density fibrinogen. Two factors may contribute to this result: engagement of GPIIb/IIIa receptors with fibrinogen on the abluminal surface leading to the loss of luminal receptors, and loss of luminal filopodia that interact with thromboerythrocytes. These data provide insight into the differences in thrombogenicity between surfaces, and may provide a mechanism for purposefully passivating platelet-reactive artificial surfaces.

Thromboerythrocytes. In vitro studies of a potential autologous, semi-artificial alternative to platelet transfusions.

In an attempt to overcome the limitations and drawbacks of using fresh platelets for transfusion therapy of thrombocytopenic patients, we have performed in vitro experiments on an autologous, semi-artificial alternative to platelet transfusions. Based on our previous studies of the interactions of unactivated and activated platelets with beads coated with peptides of various lengths, all of which contained the arginine-glycine-aspartic acid (RGD) cell recognition sequence, the peptide Ac-CGGRGDF-NH2 was chosen for covalent coupling to erythrocytes. A heterobifunctional crosslinking reagent (N-maleimido-6-aminocaproyl ester of 1-hydroxy-2-nitrobenzene-4-sulfonic acid) was used to crosslink via the peptide's free sulfhydryl group and the erythrocyte's surface amino groups. Approximately 0.5-1.5 x 10(6) peptide molecules bound per erythrocyte after 2 h of incubation, and most of the peptides appeared to crosslink to glycophorin A. The resulting cells, termed thromboerythrocytes, interacted selectively with activated platelets to form mixed aggregates. Studies with fluid phase RGD peptides and monoclonal antibodies indicated that the RGD peptides on the thromboerythrocytes interacted with the GPIIb/IIIa receptors on activated platelets. Thromboerythrocytes could also bind to platelets adherent to collagen. There was minimal erythrocyte hemolysis during the formation of thromboerythrocytes and studies of thromboerythrocyte osmotic fragility and cellular deformability showed no significant changes from control erythrocytes. Whereas there is a 20:1 ratio of erythrocytes to platelets in the circulation of normal individuals, the erythrocytes from as little as 50 ml of blood could be transformed into the equivalent of 2 U of platelets by numbers (equivalent to 18 U of platelets by mass), and reinfused into the same individual within several hours. These data encourage us to proceed to in vivo studies to assess the hemostatic efficacy of thromboerythrocytes in thrombocytopenic animals.

Clownfish in hypoxic anemones replenish host O2 at only localised scales.

The clownfish-anemone association exemplifies a symbiosis where both members benefit from nutrient exchange and protection from predators. Clownfish also perform aeration-like behaviour in their host anemones at night, but it is not yet known whether this is stimulated by the onset of hypoxia, and whether both members benefit from O2 replenishment. Oxygen at 3 distances above the sea anemone Entacmaea quadricolor (0.2, 1.2 and 2.2 cm) therefore was measured under 3 light levels (photon flux density = 0, 55 and 110 µmol m-2 s-1), with and without the anemonefish Amphiprion frenatus. Hypoxia (O2 < 50% air saturation) was recorded in the anemone, but only at 0.2 cm away from the anemone surface under dark conditions when A. frenatus was absent. This localised layer of hypoxia was eliminated by the presence of A. frenatus exhibiting aeration-like behaviour. Respirometry revealed that A. frenatus is extremely hypoxia tolerant (S crit = 14.3% at 25 °C), suggesting that aeration behaviour does not provide a major metabolic advantage to clownfish because they do not breathe water at 0.2 cm and are not metabolically constrained by O2 at distances ≥ 1.2 cm. That the aeration behaviour of A. frenatus facilitates only the metabolism of its O2-conforming host reveals a unique aspect of this symbiotic relationship.

Glutathione S-transferase mu and theta polymorphisms and breast cancer susceptibility.

BACKGROUND: The enzymes encoded by the glutathione S-transferase mu 1 (GSTM1) and theta 1 (GSTT1) genes are involved in the metabolism (mainly inactivation, but activation is possible) of a wide range of carcinogens that are ubiquitous in the environment; the enzyme encoded by the GSTT1 gene may also be active in endogenous mutagenic processes. Homozygous deletions of the GSTM1 and GSTT1 genes are commonly found in the population and result in a lack of enzyme activity. This study was undertaken to evaluate the association between GSTM1 and GSTT1 gene polymorphisms and breast cancer risk. METHODS: Our study included 466 women with incident cases of breast cancer occurring from May 1989 through May 1994 and 466 matched control subjects. These individuals were part of a prospective cohort of U.S. women (i.e., the Nurses' Health Study). Odds ratios (ORs) and 95% confidence intervals (CIs) from conditional logistic regression models were used to estimate the association between genetic polymorphisms and breast cancer risk. RESULTS: The GSTM1 and GSTT1 null genotypes were not associated with an increased risk of breast cancer (OR = 1.05 [95% CI = 0.80-1.37] for GSTM1 null; OR = 0. 86 [95% CI = 0.61-1.21] for GSTT1 null). On the contrary, a suggestion of a decreased risk of breast cancer associated with the GSTT1 null genotype was observed among premenopausal women. When considered together, no combination of the GSTM1 and GSTT1 genotypes was associated with an increased risk of breast cancer. The relationship between GSTM1 and GSTT1 gene deletions and breast cancer risk was not substantially modified by cigarette smoking. CONCLUSIONS: Our data provide evidence against a substantially increased risk of breast cancer associated with GSTM1 and/or GSTT1 homozygous gene deletions.

Glutathione S-transferase class mu deletion polymorphism and breast cancer: results from prevalent versus incident cases.

A common deletion polymorphism in the gene coding for the glutathione S-transferase class mu (the GSTM1 gene) results in a decreased ability to detoxify carcinogenic epoxide intermediates and has been associated with increased breast cancer risk in some small studies. We studied the GSTM1 gene deletion polymorphism (conferring the null genotype) in 243 women who had prevalent breast cancer and 245 women without breast cancer, who were among the 32,826 women in the Nurses' Health Study who gave a blood sample in 1989-1990. In the prevalent case series, the null genotype was slightly more common among cases (58%) than among controls (51%; age-adjusted odds ratio = 1.30; 95% confidence interval, 0.91-1.86). Among cases, the prevalence of the GSTM1 deletion increased with duration of survival [68% for > or = 8 years since diagnosis; 57% for 4-8 years; 51% for < 4 years; P (trend) = 0.04]. In an incident case series of 240 women who were diagnosed with breast cancer following blood collection and prior to June of 1992 and compared with age-matched controls, the GSTM1 deletion was not associated with an elevation in risk (relative risk, 1.08; 95% confidence interval, 0.74-1.57). No significant interaction with cigarette smoking was evident. Thus, there was no significant increase in risk of incident breast cancer associated with the GSTM1 null genotype; however, the gene deletion polymorphism appeared to confer improved survival. These data suggest that odds ratios based upon prevalent cases in molecular epidemiologic studies may be biased due to differential survival. Further studies are required to determine whether this polymorphism is associated with improved breast cancer prognosis.

Perceptual boundedness and perceptual support in conceptual development.

A theoretical framework for describing the role of perceptual information in early conceptual development is presented. In the main section of the article, a general operationalization of perceptual boundedness is introduced, 3 causes of this limitation are identified, the conditions under which infants and children seem to be perceptually bound are formulated, and the mechanisms by which this limitation declines are described. Traditional claims that young children are perceptually bound, as well as contemporary objections to these claims, are often based on the assumption that perceptual information is generally unveridical or insufficient. Recent doubts about this assumption are evaluated in the final section of the article. It is concluded that although realist arguments are untenable, there are limited forms of perceptual support for conceptual development.

Wayfinding on foot from information in retinal, not optical, flow.

People find their way through cluttered environments with ease and without injury. How do they do it? Two approaches to wayfinding are considered: Differential motion parallax (DMP) is a retinal motion invariant of near and far objects moving against fixation; the information in optical flow (IOF) is a radial pattern of vectors, relying on decomposition of retinal flow. Evidence is presented that DMP guides wayfinding during natural gait, accounting for errors as well as correct responses. Evidence against IOF is also presented, and a space-time aliasing artifact that can contaminate IOF displays is explored. Finally, DMP and IOF are separated, showing they can yield different results in different environments. Thus, it is concluded that (a) DMP and IOF are different, (b) DMP and not IOF is used for wayfinding, (c) moving observers do not usually decompose retinal flow, and (d) optical flow may be a mathematical fiction with no psychological reality.

Abortion, revised: participants in the U.S. clinical trials evaluate mifepristone.

This paper centers on the questions: How do non-surgical abortion methods affect private experiences of abortions? How might they influence public conceptions of abortion? Drawing on interviews with clients who participated in the 1994-95 U.S. clinical trials of mifepristone at one trial site (conducted during the trials), and focus group interviews conducted with health care workers at all 17 trial sites (after the trials were completed), we examine participants' evaluations of this method of abortion. Surgical abortion functioned as the reference point by which research participants assessed medical abortion. They discussed mifepristone abortion in terms of nature and invasion, privacy and bodily integrity, denial and agency. Clients frequently portrayed mifepristone abortion as a better moral choice than surgical abortion--in some cases even depicting it as not-really-an-abortion but rather as a miscarriage. Clients felt that mifepristone offered them a greater measure of control over their abortion experiences. Health care providers offered critical analysis of their clients' perceptions, yet affirmed the potential of medical abortion to offer women greater variety and latitude in procreative decision-making, and perhaps even to depoliticize the issue of abortion in the U.S. by thwarting the efforts of anti-abortionists to target providers and aborting women.

Intraosseous lipoma of the calcaneus. A case report.

The intention of this report is to heighten the practitioner's clinical suspicion of intraosseous lipoma of the calcaneus. The apparent infrequency of this diagnosis may be attributed to the dependence on radiographic evaluation and the use of closed needle biopsy. Thus, to make a proper diagnosis, a computed tomography scan with the rate of attenuation and, ultimately, a large tissue biopsy with attentive gross and microscopic examination are necessary.

Buffered lidocaine decreases the pain of digital anesthesia in the foot.

The pain associated with injections for inducing digital anesthesia can be decreased by adding sodium bicarbonate to plain lidocaine. A randomized, double-blind study has demonstrated that 24 out of 30 participants indicated on a visual analogue scale that buffered lidocaine is less painful than plain lidocaine. The pain decreased by 50% or more for almost half of the participants. Practitioners can easily buffer lidocaine in the office by adding a small volume of sodium bicarbonate to plain lidocaine.

The role of the akin osteotomy in the surgical management of hallux abducto valgus.

The Akin osteotomy has gained widespread popularity in foot surgery. This article reviews the proper indications, technique, modifications, and complications of the procedure. It also identifies the true role in the surgical management of the hallux abducto valgus deformity.

Children's awareness of the biological implications of kinship.

Preschoolers' thinking about kinship was explored by means of a simple induction task. A target animal was described as possessing a property, and children were asked whether each of 2 other animals shares the property or not. When no information about kinship was given, children in Experiment 1 based their inductions of biological properties on physical similarity. However, when kinship relations were specified, children judged that dissimilar-looking kin share more biological properties than similar-looking but unrelated members of the same species. In Experiment 2, describing the similar animals as socially related did not change the basic pattern of inductions obtained in the first experiment. Moreover, subjects in Experiment 3 did not induce more acquired physical and psychological properties among families than among unrelated animals. The results of Experiments 1 and 2 illustrate one case where young children favor a nonperceptible relation (kinship) over a perceptible one (similarity) as a basis for judgment. The overall pattern of data suggest that young children distinguish to some extent between the biological and social domains.