A novel KRT71 variant in curly-coated dogs.

Curly fur is a common phenotype in many dog breeds, known to result from a missense variant (c.451C>T) in exon 2 of the keratin 71 (KRT71) gene. During screening for this variant across various breeds, we found that Curly Coated Retrievers (CCRs) fixed with the trait did not carry the known variant. By analysis of whole-genome sequencing data of one CCR we identified a novel genetic cause for curly fur. We found a novel structural variant in exon 7 of the KRT71 gene (c.1266_1273delinsACA) that was predicted to result in a frameshift and stop loss, therefore significantly affecting the structure of the protein, if translated. The variant was also found at lower frequencies in five other breeds, including Lagotto Romagnolo, Bichon Frise, Spanish Water Dog, Chesapeake Bay Retriever and Irish Terrier. One curly-coated Lagotto carried neither of the two KRT71 variants. These results identify a second variant for curly coat in KRT71 and suggest the existence of additional alleles. This study enables the development of an additional KRT71 gene test for breeders to understand and manage coat types.

Heterozygosity testing and multiplex DNA panel screening as a potential tool to monitor health and inbreeding in a small, closed dog population.

BACKGROUND: Selective breeding in populations with a limited effective population size may result in a loss of genetic diversity, which can cause an increased concentration of specific disease liability genes. The Dutch Shepherd Dog (DSD) in the Netherlands is an example of such a breed with a small effective population. OBJECTIVE: To evaluate the measurement of genetic diversity and multiplex DNA panel screening for implementation in a breeding strategy for the Dutch Shepherd Dog (DSD) and to investigate the clinical relevance of potentially identified mutations in the multiplex DNA panel screening. RESULTS: Genome-wide SNP testing showed genetic isolation and reduced genetic diversity within coat variety subgroups of the DSD. Panel screening identified a Von Willebrand's Disease type I mutation. Although decreased Von Willebrand's Factor proteins were significantly lower in DSDs carrying the VWD-I allele compared to the wildtype, clinical follow-up did not show a significant association between the clinical phenotype and VWD-I genotype. CONCLUSIONS: Genetic relationship measurement within a breed population may be a useful tool to enable breeding strategies to conserve genetic diversity. Results from a disease panel screening need to be evaluated for clinical relevance before breed selection restrictions can be considered.

Founder representation and effective population size in old versus young breeds-genetic diversity of Finnish and Nordic Spitz.

Finnish Spitz is 130-year-old breed and has been highly popular in Finland throughout its history. Nordic Spitz is very similar to Finnish Spitz by origin and use, but is a relatively recent breed with much smaller population size. To see how breed age and breeding history have influenced the current population, we performed comprehensive population genetic analysis using pedigree data of 28,119 Finnish and 9,009 Nordic Spitzes combined with genomewide single nucleotide polymorphism (SNP) data from 135 Finnish and 110 Nordic Spitzes. We found that the Finnish Spitz has undergone repeated male bottlenecks resulting in dramatic loss of genetic diversity, reflected by 20 effective founders (fa ) and mean heterozygosity (Hz) of 0.313. The realized effective population size in the breed based on pedigree analysis (N¯ec) is 168, whereas the genetic effective population size (Neg ) computed the decay of linkage disequilibrium (r2 ) is only 57 individuals. Nordic Spitz, although once been near extinction, has not been exposed to similar repeated bottlenecks than Finnish Spitz and had fa of 27 individuals. However, due to the smaller total population size, the breed has also smaller effective population size than Finnish Spitz (N¯ec = 98 and Neg  = 49). Interestingly, the r2 data show that the effective population size has contracted dramatically since the establishment of the breed, emphasizing the role of breed standards as constrains for the breeding population. Despite the small population size, Nordic Spitz still maintains SNP heterozygosity levels similar to mixed breed dogs (mean Hz = 0.409). Our study demonstrates that although pedigree analyses cannot provide estimates of the present diversity within a breed, the effective population sizes inferred from them correlate with the genotyping results. The genetic relationships of the northern Spitz breeds and the benefits of the open breed registry are discussed.

Characteristics of a 128 x 128 liquid-crystal spatial light modulator for wave-front generation.

Spatial and temporal characteristics of a 128x128 zero-twist nematic liquid-crystal spatial light modulator are investigated for wave-front generation at a wavelength of 632.8 nm. The modulator is found to have the capability of producing at least eight phase levels between 0 and 2pi , and the rate of arbitrary phase modulation is limited to approximately 4.5 Hz. Wave-front generation of the first 55 Zernike polynomials is demonstrated.

[Nutrition education --an important matter in medical education]. / Näringslära--angeläget ämne i läkarutbildningen.

At the Nordic Federation of Medical Education congress on nutrition held at Lund in April 1991, it was agreed that nutrition should be allotted adequate space within the framework both of the preclinical and clinical curricula. A proposal was presented whereby a basic course in nutrition would be scheduled early in the general medical curriculum, together with clinical nutrition both integrated with other suitable clinical subjects and as a self-contained subject. The physician's role in a preventive capacity in the public health sector was discussed, as were research aspects of the subject.

Automated measurement of lipoprotein(a) by immunoturbidimetric analysis.

Immunoturbidimetric analysis of lipoprotein(a) in plasma or serum was developed for use on the Roche COBAS FARA II and COBAS MIRA clinical chemistry analyzers. The components of the assay are: (1) buffer consisting of 2.25% polyethylene glycol in phosphate-buffered saline, 0.2% gelatin, and a surfactant; (2) fractionated goat anti-human lipoprotein(a) IgG; (3) five standards with lipoprotein(a) concentrations ranging from 0.05 to 1.0 g/l; (4) two controls with concentrations of approximately 0.2 and 0.5 g/l. The analyzer delivers sample and buffer, incubates the reaction mixture at 37 degrees C for 5 min, delivers neat lipoprotein(a) antibody, and incubates for an additional 10 min. The lipoprotein(a) concentration of samples is calculated by the COBAS DENS (Data Evaluation for Non-linear Standard Curves) option by fitting the standard curve values to a four-parameter logit-log curve model. Total imprecision results (CV%) for the FARA II and MIRA were under 11% (NCCLS protocol EP5-T). The assay is linear beyond the highest calibrator to 2.6 g/l. No interference was observed for plasminogen up to 2.3 g/l, apolipoprotein B up to 4.36 g/l, hemoglobin up to 10 g/l, bilirubin up to 4.0 g/l, and triglycerides up to 4.36 g/l. Comparison with a double monoclonal ELISA used at the Northwest Lipid Research Laboratories yielded: R = 0.970, slope = 1.013, and y-intercept = 0.00009 (n = 37). Comparison with a commercially available ELISA kit for lipoprotein(a) yielded: r = 0.987, slope = 1.243, and y-intercept = 0.024 (n = 40). This assay provides rapid, accurate, and precise screening of lipoprotein(a) in serum or plasma.

Findings from the preparticipation athletic examination and athletic injuries.

This study investigated the relationships between the findings from a standardized preparticipation athletic examination, the sport played, and athletic injuries requiring treatment by a physician and/or requiring the athlete to miss one or more games. Of public high school students receiving a preparticipation athletic examination during the 1989-1990 academic year, 674 (56%) either completed a telephone interview or returned a mailed questionnaire at the end of the academic year. The sample consisted of 408 (60.5%) blacks and 243 (36.1%) whites; 470 (69.7%) of the subjects were males. The subjects ranged in age from 13 to 20 years (mean +/- SD, 16.1 +/- 1.2 years), and participated in at least 10 school sports. Injuries were reported by 29.5% of the athletes. The highest proportion of athletes injured occurred among male football (36.3%), female basketball (33.3%), male baseball (19.4%), male soccer (17.2%), and female track and field (15.8%) participants. Responses by the athletes and their parents on the standardized health history were significantly associated with injuries in several specific areas. Knee injuries were associated with previous knee injuries, knee surgery, and history of injuries requiring medical treatment. Ankle injuries were associated with previous ankle injuries and previous injuries requiring medical treatment. Both arm and other leg injuries were associated with previous fractures. Male athletes with either abnormal knee or ankle findings from the physical examination were more likely to injure the knee or ankle, respectively. However, the sensitivities and positive predictive values of these relationships are weak. These data suggest that the preparticipation athletic examination may not predict certain athletic injuries and that additional prevention efforts for specific body areas of injury are needed in certain sports.

Adolescents' attrition from school-sponsored sports.

This study investigated reasons for adolescents' attrition from school-sponsored sports teams and the relationship between attrition and intention to participate in organized and nonorganized sports in the future. Of 674 high school student athletes, 26% dropped off at least one team in 1989-1990, and overall attrition increased to 29.8% when attrition from more than one sport was considered. Attrition was higher among black students, students reporting an athletic injury, those injured playing football, and those sustaining a knee injury. Injury was cited as the most frequent reason for attrition, followed by being cut from the team, "other reasons," needing to get a job, inconvenient game or practice schedule, and needing more time to study, although the main reason for attrition also varied by sport. When athletes were classified according to externally precipitated attrition, athlete-initiated attrition, or completion of the sports season, no differences were noted in the three groups' intent to engage in organized sports in the future. However, the group with externally precipitated attrition expressed a stronger intent to engage in nonorganized sports in the future than the other two groups.

Alteration of proteoglycan metabolism during the differentiation of 3T3-L1 fibroblasts into adipocytes.

3T3-L1 fibroblasts were induced to differentiate to 3T3-L1 adipocytes by dexamethasone, isobutyl-methylxanthine, and insulin. To study how differentiation affects extracellular matrix production, the accumulation of proteoglycans was studied by labeling the 3T3-L1 cells with [35S]sulphate for 24 h. The labeled proteoglycans were isolated from the medium and cell layer extracts by anion-exchange chromatography. They were then taken to gel filtration chromatography on Superose 6 before or after chondroitin ABC lyase digestion. Hyaluronan was determined by radioimmunoassay. The rate of accumulation of proteoglycans and hyaluronan in the control 3T3-L1 fibroblasts increased with time whereas it decreased slightly in the age matched adipocytes where the differentiation had proceeded, as judged by the change of morphology and increase of the activity of the adipose conversion markers glycerol-3-phosphate dehydrogenase and hormone sensitive lipase. The main change noted was that the adipocytes accumulated 50-70% less amount of small proteoglycans (decorin) in the medium than the fibroblasts did. The amount of large chondroitin/dermatan sulphate proteoglycans was also decreased but to a considerably smaller extent (30%). In the cell layer, heparan sulphate proteoglycan decreased by 60% as compared with the control cells. Thus, the differentiation of 3T3-L1 fibroblasts into adipocytes, which changes the morphology and the function of the cells, is also accompanied by a decreased net production especially of proteoglycans typical of fibrous connective tissue.

Growth hormone increases phosphoinositide turnover in rat adipocytes that are sensitive to the insulin-like action of the hormone.

UNLABELLED: The effect of pituitary human growth hormone (hGH) on the 32P-labelling of phosphoinositides and phosphatidic acid was studied in noradrenaline-stimulated rat adipocytes which were either responsive or non-responsive to the antilipolytic (insulin-like) effect of hGH. In cells responsive to the insulin-like effect of hGH, hormone treatment resulted in a marked increase of the 32P-labelling of phosphatidic acid and phosphatidyl inositol in the plasma membrane, high density microsomal, and low density microsomal fractions. The increased 32P-labelling most likely reflects an activation of phosphoinositide phospholipase C. ABBREVIATIONS: IP3 - Myo-Inositol 1,4,5-Triphosphate. BSA - Bovine Serum Albumin, HEPES - N-2-Hydroxyethylpiperazine-N'-2-Ethanesulphonic Acid, hGH - Human Pituitary Growth Hormone, LPC-Lysophosphatidylcholine, LPE - Lysophosphatidylethanolamine, LPI - Lysophosphatidylinositol, PA - Phosphatidic Acid, PC - Phosphatidylcholine, PE - Phosphatidylethanolamine, PI - Phosphatidylinositol, PIP - Phosphatidylinositol-4-Phosphate, PIP2-Phosphatidylinositol-4,5-Diphosphate, PS-Phosphatidylserine.

Monoclonal antibodies to the pituitary growth-hormone receptor by the anti-idiotypic approach. Production and initial characterization.

We obtained 10/192 and 3/384 antibody-secreting hybrids after immunization of Balb/c mice with either human growth hormone or affinity-purified rabbit anti-(human growth hormone) respectively. Radiolabelled rabbit anti-(human growth hormone) antibodies, but not human growth hormone, were specifically bound by supernatants from the 13 hybrids. The binding was completely inhibited by human-growth-hormone serum binding protein. However, anti-(human growth hormone antibodies) were detected in the sera of all the mice immunized with human growth hormone. In an independent fusion, which was carried out after immunization with fewer doses of human growth hormone, anti-(human growth hormone) antibodies were also obtained. Five hybrids, where the starting antigen was human growth hormone, were selected for ascites production, and the corresponding monoclonal antibodies were partially purified and characterized with respect to their immunoglobulin isotype and their interaction with human-growth-hormone receptors. These antibodies were found to enhance the binding of radioiodinated human growth hormone to human-growth-hormone serum binding protein from human and rabbit plasma by 40%. Scatchard analysis of the effect of one of the monoclonal antibodies showed that this enhancement was due to an increased number of binding sites. All of the partially purified antibodies but one (F12) inhibited the binding of human growth hormone to rat but not rabbit, liver microsomes to various extents, as well as to H-4-II-E rat hepatoma cells. Monoclonal antibody F12 enhanced the binding of radiolabelled human growth hormone to rat liver microsomes and H-4-II-E hepatoma cells. This enhancement was found to be due to an increase in the number of binding sites.

The acute GH action in rat adipocytes is associated with enhanced phosphorylation of a 46 kDa plasma membrane protein enriched by GH-Sepharose.

The acute anti-lipolytic effect of human growth hormone (hGH) in maximally noradrenaline-stimulated intact rat adipocytes was selectively associated with increased phosphorylation of a 46 kDa plasma membrane protein which was highly enriched by hGH-Sepharose chromatography. The same protein was also phosphorylated by an endogenous protein kinase in isolated plasma membranes, although then no hGH effect could be demonstrated. About 14% of the phosphate incorporated into the protein in isolated plasma membranes was found in tyrosine residues and the remainder in serine and threonine. The possible relation of the 46 kDa protein with the hGH plasma membrane receptor is discussed.

An assessment of prescribing using drug utilization review criteria.

Drug utilization review screening criteria were applied to a sample of 30 000 prescriptions written by Iowa physicians. Characteristics of the physicians who had high percentages of prescriptions violating the explicit criteria were examined. It was found that the proportion of prescriptions failing the screening criteria did not differ significantly among physicians of differing board certification groups, medical school of graduation, year of graduation from medical school, age, or size of town where the physicians practiced. Regardless of the physicians' demographic characteristics, approximately 50 percent of their prescriptions violated the prescribing criteria. The three categories of screening criteria included irrational mixture, quantity prescribed, and daily dosage. An analysis of the relationship between physician ratings obtained for each of the categories revealed that no relationship existed between a physician's quantity prescribed rating and daily dosage rating, or between a physician's irrational mixture rating and daily dosage rating. A negative relationship was found between a physician's quantity prescribed rating and irrational mixture rating. This tends to indicate that the three categories measure different prescribing patterns and each warrants respective monitoring by pharmacists.

A calorimetric investigation of the interaction of the lac repressor with inducer.

A calorimetric study has been made of the interaction between the lac repressor and isopropyl-1-thio-beta-D-galactopyranoside (IPTG). The buffer-corrected enthalpy of reaction at 25 degrees C was found to be -15.6, -24.7, -4.6 kJ/mol of bound IPTG at pH 7.0, pH 8.1, and pH 9.0, respectively. This large range of enthalpy values is in contrast to a maximum difference in the free energy of the reaction of only 1.5 kJ/mol of bound IPTG between these pH values. The reaction was found by calorimetric measurements in different buffers to be accompanied by an uptake of 0.29 mol of protons/mol of bound IPTG at pH 8.1. The pH dependency of the reaction enthalpy suggests differences in the extent of protonation of the binding site and the involvement of H bonding with IPTG. The lack of strong hydrophobic contributions in the IPTG binding process is revealed by the absence of any determinable heat capacity change for the reaction at pH 7.0. The presence of phosphate buffer significantly alters the enthalpy of IPTG binding at higher pH values, but has little effect upon the binding constant. This implies that highly negative phosphate species change the nature of the IPTG binding site without any displacement of phosphate upon IPTG binding.