The assessment of genetic diversity within and among the eight subpopulations of Japanese Black cattle using 52 microsatellite markers.

Japanese Black cattle are at risk for genetic homogeneity due to intensive use of a few sires. Therefore, assessment of the actual genetic diversity of this breed is important for future breeding plans. In the present study, we investigated the genetic diversity within and among eight subpopulations of Japanese Black cattle using 52 microsatellite markers. The parameters for genetic diversity of Japanese Black cattle were comparable to those of other cattle breeds, suggesting that the relatively high genetic diversity of the breed. However, upon comparison among the eight subpopulations, the Hyogo subpopulation showed markedly low genetic diversity. The results of the pairwise FST values, phylogenetic network and structure analysis indicated that the Hyogo population has remarkably high level of genetic differentiation from other populations, while Yamagata, Niigata, Hiroshima and Kagawa populations have low levels of genetic differentiation. Furthermore, multidimensional scaling plots indicated that individuals in some subpopulations were separated from individuals in the other subpopulations. We conclude that while the overall genetic diversity of Japanese Black cattle is still maintained at a relatively high level, that of a particular subpopulation is significantly reduced, and therefore the effective population size of the breed needs to be controlled by correct mating strategies.

A novel mutation of the bovine EDA gene associated with anhidrotic ectodermal dysplasia in Holstein cattle.

Anhidrotic ectodermal dysplasia (EDA) is a genetic disease characterized by the absence or hypoplasia of hair, teeth and eccrine sweat glands that has been reported in humans, the tabby mouse mutants, cattle and dogs. The EDA gene on the X chromosome encodes a protein, ectodysplasin-A (EDA), which is responsible for EDA. Here we describe a novel mutation of the EDA gene in which a 19 bp deletion in exon 1 in male Holstein calves demonstrated the phenotypic features of EDA. The dam and the grand-dam of the affected calves were heterozygous for this deletion. It is assumed that this deletion close to the start codon confuses all transcripts, and leads to the complete loss of pleiotropic functions of the bovine EDA gene. These results suggest that this mutation might be useful as animal models for the investigation of the pathogenic mechanisms of the anhidrotic ectodermal dysplasia.

The diversity of bovine MHC class II DRB3 and DQA1 alleles in different herds of Japanese Black and Holstein cattle in Japan.

In cattle, bovine leukocyte antigens (BoLAs) have been extensively used as markers for bovine diseases and immunological traits. In this study, we sequenced alleles of the BoLA class II loci, BoLA-DRB3 and BoLA-DQA1, from 650 Japanese cattle from six herds [three herds (507 animals) of Japanese Black cattle and three herds (143 animals) of Holstein cattle] using polymerase chain reaction-sequence-based typing (PCR-SBT) methods. We identified 26 previously reported distinct DRB3 alleles in the two populations: 22 in Japanese Black and 17 in Holstein. The number of DRB3 alleles detected in each herd ranged from 9 to 20. Next, we identified 15 previously reported distinct DQA1 alleles: 13 in Japanese Black and 10 in Holstein. The number of alleles in each herd ranged from 6 to 10. Thus, allelic divergence is significantly greater for DRB3 than for DQA1. A population tree on the basis of the frequencies of the DRB3 and DQA1 alleles showed that, although the genetic distance differed significantly between the two cattle breeds, it was closely related within the three herds of each breed. In addition, Wu-Kabat variability analysis indicated that the DRB3 gene was more polymorphic than the DQA1 gene in both breeds and in all herds, and that the majority of the hypervariable positions within both loci corresponded to pocket-forming residues. The DRB3 and DQA1 heterozygosity for both breeds within each herd were calculated based on the Hardy-Weinberg equilibrium. Only one Japanese Black herd showed a significant difference between the expected and observed heterozygosity at both loci. This is the first report presenting a detailed study of the allelic distribution of BoLA-DRB3 and -DQA1 genes in Japanese Black and Holstein cattle from different farms in Japan. These results may help to develop improved livestock breeding strategies in the future.

Anterior crossbite in the primary dentition: proposal for a new analytical method in children.

The aim of this study was to clarify the features of profiles of patients with anterior crossbite in early childhood, using a new method of analysis. Lateral cephalograms of 139 (68 males; 71 females) untreated child patients with anterior crossbite in the primary dentition were assessed. The patients were aged from 3 to 5 years. To evaluate disharmony between the maxilla and the mandible, a proprietary calculating system, the Theoretical Individualized Profile drawing System (TIPS), was applied. Using this system, individualized standard cephalometric values were obtained for normal occlusion. This standard profile, which was harmonized with the cranial base structure of each patient, was compared with the child's original profile. The sizes of the mandibles in the children with anterior crossbite, subclassified into three age groups (3, 4, and 5 years) were greater, both horizontally and vertically, than those in the mean profiles derived from TIPS. The vertical size of the maxilla was smaller than that derived from TIPS in the patients aged 3 years. In contrast, in the patients aged 5 years, the vertical size of the maxilla was larger than that derived from TIPS. These results suggest that, in children with anterior crossbite, the vertical growth of the maxilla is closely related to the variation of morphological conditions with development.

Crystallization and preliminary X-ray analysis of Thermoactinomyces vulgaris R-47 maltooligosaccharide-metabolizing enzyme homologous to glucoamylase.

A maltooligosaccharide-metabolizing enzyme from Thermoactinomyces vulgaris R-47 (TGA) homologous to glucoamylase degrades maltooligosaccharides more efficiently than starch, unlike fungal glucoamylases. TGA was crystallized and the state of the protein in solution was analyzed by gel-filtration chromatography. Diffraction data were collected to 3.31 A resolution. The TGA crystal belongs to the orthorhombic space group P2(1)2(1)2(1) or P2(1)2(1)2, with unit-cell parameters a = 110.2, b = 317.6, c = 144.9 A, and is expected to contain five to eight TGA molecules per asymmetric unit. Gel-filtration and native PAGE analyses indicated that TGA exists as a dimer in solution. This is the first report of the crystallization of an oligomeric glucoamylase.

Genotype of stearoyl-coA desaturase is associated with fatty acid composition in Japanese Black cattle.

To investigate the genetic factors that affect fatty acid composition of beef, we compared the full-length bovine stearoyl-CoA desaturase (SCD) cDNA from 20 Japanese Black steers. Two types of the SCD gene with single nucleotide polymorphisms (SNPs) were observed in the ORF of SCD cDNA, in which an amino acid replacement from valine (type V) to alanine (type A) was predicted. We developed a method for genotyping these two SCD genes based on PCR-RFLP. We have classified 1003 Japanese Black carcasses into three genotypes, VV, VA, and AA, and compared fatty acid composition among them. The SCD type A gene contributed to higher MUFA percentage and lower melting point in intramuscular fat. The SCD genotype was not the only genetic factor contributing to fatty acid composition of Japanese Black steers, but the SCD genotype was considered one of the causes of genetic variation in fatty acid composition of Japanese Black steers. Transcription factors such as sterol regulatory element binding protein-1c (SREBP-1c) may account for the remaining part of the genetic variation in fatty acid composition.

Congenital defect of maxillary primary central incisor associated with exposed pulp and gingival [fibrosis]: case report.

This report describes a rare case of hypoplastic primary incisor in which the pulp was exposed at the crown portion and covered by the gingiva in a 1-year-11-month-old boy. The patient was referred to us due to swelling of his labial cervical gingiva of the maxillary right primary central incisor, and on examination, extended to the hypoplastic labial surface. Radiographically, there was a round radiolucent area on the crown including the edge. Surgical removal of the swollen gingiva revealed a large defect of the labial aspect of the incisor, showing pulpal tissue inside. The tooth was treated by vital pulpotomy. Histopathologically, the removed gingival tissue contained many pieces of dysplastic tooth elements in the lamina propria portion which should have been connected to the exposed pulp. The findings suggested that pulp exposure resulted from focal dental hypoplasia not from resorption of the tooth.

(eta 5-Cyclopentadienyl)(eta 4-di- and tetra-phosphorylcyclobutadiene)cobalt(I): synthesis, structure, and formation of 1-D coordination polymer.

(eta 5-Cyclopentadienyl)(eta 4-di- and tetra-phosphorylcyclobutadiene)cobalt(I) complexes were synthesized by the reaction of mono- and diphosphorylacetylenes with CpCo(CO)2, respectively. The tetraphosphoryl derivative has proved to work as a bis-bidentate ligand affording a one-dimensional coordination polymer with Ce(III).

Relationship between cranial base structure and maxillofacial components in children aged 3-5 years.

The aim of this study was to clarify the inter-relationships between cranial base angle, the morphological variations of maxillofacial components, and growth of the anterior cranial base length. One hundred and twenty-two Japanese children aged 3-5 years with normal occlusion in the primary dentition were included in this investigation. To analyse the relationship between cranial base structure and maxillofacial components, the subjects were divided into three groups according to facial type (prognathic, orthognathic, or retrognathic) assessed by the magnitude of their NSAr angle (as cranial base angle). These categorized cephalometric profiles suggested that the antero-posterior location of the maxillofacial components corresponded to the NSAr angle. Factor analysis of cephalometric variables showed that the variation of maxillofacial components from the S-N line was strongly related to the cranial base angle. This normal range of morphological variation was distinguished from that of growth by the factor loadings. From these results, theoretical models of cephalometric profiles with normal occlusion were computed by linear regression analysis. Using the models, a standard profile related to an arbitrary cranial base angle was obtained for children with normal occlusion, aged 3-5 years.

Chylomicronemia caused by lipoprotein lipase gene mutation related to a hyper-response of insulin secretion to glucose.

A 39-year-old man with lipoprotein lipase (LPL) deficiency (height 177.7 cm, body weight 67 kg, and body mass index 21.2 kg/m2) showed severe hypertriglyceridemia (2,032 mg/dl). LPL activity and concentration were markedly low in postheparin plasma. LPL gene analysis revealed a homozygous mutation, Asp204 --> Glu in exon 5. Fasting plasma glucose (81 mg/dl) and insulin (2.7 microU/ml) levels were normal. Plasma glucose pattern during oral glucose (75 g) tolerance test was normal, however 30 minutes after glucose-loading the insulin secretion unexpectedly increased to 89.4 microU/ml. These data suggested that chylomicronemia might be related to a hyper-response of insulin secretion to glucose without obesity.