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Interspecies somatic cell nuclear transfer (iSCNT) has an immense potential to rescue endangered animals and extinct species like mammoths. In this study, we successfully established an Asian elephant’s fibroblast cell lines from ear tissues, performed iSCNT with porcine oocytes and evaluated the in vitro and in vivo development of reconstructed embryos. A total of 7780 elephant–pig iSCNT embryos were successfully reconstructed and showed in vitro development with cleavage rate, 4-cell, 8-cell and blastocyst rate of 73.01, 30.48, 5.64, and 4.73%, respectively. The total number of elephant–pig blastocyte cells and diameter of hatched blastocyte was 38.67 and 252.75 μm, respectively. Next, we designed species-specific markers targeting EDNRB, AGRP and TYR genes to verify the genome of reconstructed embryos with donor nucleus/species. The results indicated that 53.2, 60.8, and 60.8% of reconstructed embryos (n = 235) contained elephant genome at 1-cell, 2-cell and 4-cell stages, respectively. However, the percentages decreased to 32.3 and 32.7% at 8-cell and blastocyst stages, respectively. Furthermore, we also evaluated the in vivo development of elephant–pig iSCNT cloned embryos and transferred 2260 reconstructed embryos into two surrogate gilts that successfully became pregnant and a total of 11 (1 and 10) fetuses were surgically recovered after 17 and 19 days of gestation, respectively. The crown-rump length and width of elephant–pig cloned fetuses were smaller than the control group. Unfortunately, none of these fetuses contained elephant genomes, which suggested that elephant embryos failed to develop in vivo. In conclusion, we successfully obtained elephant–pig reconstructed embryos for the first time and these embryos are able to develop to blastocyst, but the in vivo developmental failure needs further investigated.

The neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) have been reported to be a predictor for intravenous immunoglobulin (IVIG) resistance in patients with Kawasaki disease (KD) recently. The objective of the present study was to elucidate the predictive validity of this new marker in a multicenter study. We retrospectively reviewed the clinical records of 520 consecutive KD patients (development data set) and 332 subsequent patients (validation data set) at 7 hospitals in Japan. Both NLR and PLR were significantly higher in the IVIG-resistant group than in the IVIG-responsive group. When we set the cut-off point as NLR ≥ 4.11 and PLR ≥ 119, multiple logistic regression analyses showed that a high NLR and PLR before initial IVIG were independent predictors of IVIG resistance, and their combination was a stronger predictor than either alone. The sensitivity and specificity of the combination of NLR ≥ 4.11 and PLR ≥ 119 were 0.58 and 0.73 in the development data set. Validated using an independent data set, they were 0.54 and 0.72 in the validation data set. On comparing the AUC of this predictor with those of the Gunma and Kurume scores, the AUC was highest for this predictor, followed by the Gunma score and Kurume score (0.70, 0.68, and 0.64, respectively). The predictive validity of the combination of a high NLR and PLR, which is a simple and convenient indicator, was equal to or better than that of the existing scoring systems. The new predictive marker may be a suitable indicator for predicting IVIG resistance in KD patients.

The Creeper (Cp) chicken is characterized by chondrodystrophy in Cp/+ heterozygotes and embryonic lethality in Cp/Cp homozygotes. However, the genes underlying the phenotypes have not been fully known. Here, we show that a 25 kb deletion on chromosome 7, which contains the Indian hedgehog (IHH) and non-homologous end-joining factor 1 (NHEJ1) genes, is responsible for the Cp trait in Japanese bantam chickens. IHH is essential for chondrocyte maturation and is downregulated in the Cp/+ embryos and completely lost in the Cp/Cp embryos. This indicates that chondrodystrophy is caused by the loss of IHH and that chondrocyte maturation is delayed in Cp/+ heterozygotes. The Cp/Cp homozygotes exhibit impaired DNA double-strand break (DSB) repair due to the loss of NHEJ1, resulting in DSB accumulation in the vascular and nervous systems, which leads to apoptosis and early embryonic death., Kinoshita et al find that the classical Creeper (Cp) phenotype in chicken is caused by a deletion containing not only the gene encoding Indian hedgehog, previously implicated in the Cp trait, but also the NHEJ1 gene encoding a DNA repair factor. They show that early death in Cp/Cp chicken is caused by impaired DNA repair and abnormalities of the vascular and nervous systems.

The precursor of heme, protoporphyrin IX (PPIX), accumulates abundantly in the uteri of birds, such as Japanese quail, Coturnix japonica, which has brown-speckled eggshells; however, the molecular basis of PPIX production in the uterus remains largely unknown. Here, we investigated the cause of low PPIX production in a classical Japanese quail mutant exhibiting white eggshells by comparing its gene expression in the uterus with that of the wild type using transcriptome analysis. We also performed genetic linkage analysis to identify the causative genomic region of the white eggshell phenotype. We found that 11 genes, including 5’-aminolevulinate synthase 1 (ALAS1) and hephaestin-like 1 (HEPHL1), were specifically upregulated in the wild-type uterus and downregulated in the mutant. We mapped the 172 kb candidate genomic region on chromosome 6, which contains several genes, including a part of the paired-like homeodomain 3 (PITX3), which encodes a transcription factor. ALAS1, HEPHL1, and PITX3 were expressed in the apical cells of the luminal epithelium and lamina propria cells of the uterine mucosa of the wild-type quail, while their expression levels were downregulated in the cells of the mutant quail. Biochemical analysis using uterine homogenates indicated that the restricted availability of 5’-aminolevulinic acid is the main cause of low PPIX production. These results suggest that uterus-specific transcriptional regulation of heme-biosynthesis-related genes is an evolutionarily acquired mechanism of eggshell pigment production in Japanese quail. Based on these findings, we discussed the molecular basis of PPIX production in the uteri of Japanese quails.

Birds display various body colors and patterns; some of them exhibit very beautiful colors and complex patterns. The main part in birds that shows body color is feathers. There are three known systems for the expression of plumage colors: (1) eumelanin and pheomelanin produced by integumental melanocytes, (2) accumulation of dietary pigments such as carotenoids, pterin, and psittacofulvins, from foods; and (3) structural colors as a result of fine structure of feathers and without special pigments. The pigment cell type in the trunk region of birds is the only melanocyte that produces melanin (eumelanin and pheomelanin), which is the same as in mammals. Since birds have additional two mechanisms (2 and 3) as noted above, they exhibit colorful body colors that are not shown in mammals. Based on the contents and ratio of eumelanin and pheomelanin, the feathers exhibit white, orange, yellow, brown, gray, or black colors. Other colors such as bright yellow, red, or glossy metallic colors of red, blue, green, or purple, are derived by the second and third mechanisms. The body and feather color patterns and color combinations are determined in the feather root, during the feather developmental process. Body colors and patterns in birds often exhibit differences in body parts, age, sex, and season. Although during evolution, birds lost the colorful erythrophore, xanthophore, and iridophores found in lower vertebrates, they obtain dietary pigments from foods and show structural color due to the fine structure of feathers. With these colorations, feathers in birds often show amazing complex patterns. The diversity of body colors and patterns in birds are very important for their survival strategy.

Background Febrile urinary tract infection (fUTI) is the most common serious bacterial infection in children. Despite this, there have been no studies examining the clinical features of pediatric fUTI in Japan. The purpose of this study was to describe the clinical characteristics of fUTI in Japanese children. Methods A multicenter, retrospective, observational study was conducted at 21 hospitals in Japan. Children under the age of 15 years who were diagnosed with fUTI between 2008 and 2017 were included. The diagnostic criteria were a temperature over 38 °C and the presence of a single bacterial pathogen in urine culture. Patient characteristics were obtained from medical records. Results In total, 2,049 children were included in the study. The median age was 5 months, and 59.3% were male. It was found that 87.0% of the males and 53.2% of the females were under 1 year of age. The main causative pathogens identified were Escherichia coli and Enterococcus spp., accounting for 76.6% and 9.8% of infections, respectively. Conclusions There was a male predominance of fUTI in Japanese children, particularly in infants. Enterococcus spp. were the second most frequent causative pathogen; therefore, Gram staining of urine samples is strongly recommended before initiating antibiotic therapy.

The genetic characteristics and diversity of 21 experimental chicken lines registered with the National BioResource Project of Japan were examined using mitochondrial D-loop sequences and 54 microsatellite DNA markers. A total of 12 haplotypes were detected in the 500-bp mitochondrial DNA sequences of the hypervariable segment I for 349 individuals of 21 lines. The 12 haplotypes belonged to three (A, D, and E) haplogroups, out of the eight (A‒H) common haplogroups in domestic chickens and red junglefowls. The haplogroups A and D were widely represented in indigenous chickens in the Asian and Pacific regions, and the haplogroup E was the most prevalent in domestic chickens. Genetic clustering by discriminant analysis of principal components with microsatellite markers divided 681 individuals of 21 lines into three groups that consisted of Fayoumi-, European-, and Asian- derived lines. In each of the cladograms constructed with Nei’s genetic distances based on allele frequencies and the membership coefficients provided by STRUCTURE and with the genetic distance based on the proportion of shared alleles, the genetic relationships coincided well with the breeding histories of the lines. Microsatellite markers showed remarkably lower genetic heterozygosities (less than 0.1 observed heterozygosity) for eight lines (GSP, GSN/1, YL, PNP, BM-C, WL-G, BL-E, and #413), which have been maintained as closed colonies for more than 40 years (except for #413), indicating their usefulness as experimental chicken lines in laboratory animal science research.

The mutant plumage color "extended brown (EB)" of the blue-breasted quail was genetically investigated. Mating experiments revealed that the EB plumage is controlled by an autosomal, incompletely dominant allele, for which we propose the symbol Eb. The EB plumage is characterized by dark brown color, and homozygotes for this mutation generally showed darker pigmentation than the heterozygotes. DNA sequencing and PCR-RFLP analyses of the EB mutants showed a rigid association between the EB plumage and a G-to-A nucleotide substitution at position 274 in the melanocortin 1-receptor gene (MC1R), clearly indicating that MC1R is the candidate gene for the EB plumage color in the blue-breasted quail.

Hybrid incompatibility, such as sterility and inviability, prevents gene flow between closely-related populations as a reproductive isolation barrier. F1 hybrids between chickens and Japanese quail (hereafter, referred to as quail), exhibit a high frequency of developmental arrest at the preprimitive streak stage. To investigate the molecular basis of the developmental arrest at the preprimitive streak stage in chicken-quail F1 hybrid embryos, we investigated chromosomal abnormalities in the hybrid embryos using molecular cytogenetic analysis. In addition, we quantified gene expression in parental species and chicken- and quail-derived allele-specific expression in the hybrids at the early blastoderm and preprimitive streak stages by mRNA sequencing. Subsequently, we compared the directions of change in gene expression, including upregulation, downregulation, or no change, from the early blastoderm stage to the preprimitive streak stage between parental species and their hybrids. Chromosome analysis revealed that the cells of the hybrid embryos contained a fifty-fifty mixture of parental chromosomes, and numerical chromosomal abnormalities were hardly observed in the hybrid cells. Gene expression analysis revealed that a part of the genes that were upregulated from the early blastoderm stage to the preprimitive streak stage in both parental species exhibited no upregulation of both chicken- and quail-derived alleles in the hybrids. GO term enrichment analysis revealed that these misregulated genes are involved in various biological processes, including ribosome-mediated protein synthesis and cell proliferation. Furthermore, the misregulated genes included genes involved in early embryonic development, such as primitive streak formation and gastrulation. These results suggest that numerical chromosomal abnormalities due to a segregation failure does not cause the lethality of chicken-quail hybrid embryos, and that the downregulated expression of the genes that are involved in various biological processes, including translation and primitive streak formation, mainly causes the developmental arrest at the preprimitive streak stage in the hybrids.

Organic ferromagnets and advanced magneticmetals have received continued interest over many decades. The systematic investigation of open-shell radicals had been reported by Morita and co-workers. ...

Japanese indigenous chickens have a long breeding history, possibly beginning 2000 years ago. Genetic characterization of Japanese indigenous chickens has been performed using mitochondrial D-loop region and microsatellite DNA markers. Their phylogenetic relationships with chickens worldwide and genetic variation within breeds have not yet been examined. In this study, the genetic characteristics of 38 Japanese indigenous chicken breeds were assessed by phylogenetic analyses of mitochondrial D-loop sequences compared with those of indigenous chicken breeds overseas. To evaluate the genetic relationships among Japanese indigenous chicken breeds, a STRUCTURE analysis was conducted using 27 microsatellite DNA markers. D-loop sequences of Japanese indigenous chickens were classified into five major haplogroups, A&ndash, E, among 15 haplogroups found in chickens worldwide. The haplogroup composition suggested that Japanese indigenous chickens originated mainly from China, with some originating from Southeast Asia. The STRUCTURE analyses revealed that Japanese indigenous chickens are genetically differentiated from chickens overseas, Japanese indigenous chicken breeds possess distinctive genetic characteristics, and Jidori breeds, which have been reared in various regions of Japan for a long time, are genetically close to each other. These results provide new insights into the history of chickens around Asia in addition to novel genetic data for the conservation of Japanese indigenous chickens.

The L strain of Japanese quail exhibits a plumage phenotype that is light yellowish in colour. In this study, we identified a nonsense mutation in the premelanosome protein (PMEL) gene showing complete concordance with the yellowish plumage within a pedigree as well as across strains by genetic linkage analysis of an F2 intercross population using approximately 2,000 single nucleotide polymorphisms (SNPs) that were detected by double digest restriction site-associated DNA sequencing (ddRAD-seq). The yellowish plumage was inherited in an autosomal recessive manner, and the causative mutation was located within an 810-kb genomic region of the LGE22C19W28_E50C23 linkage group (LGE22). This region contained the PMEL gene that is required for the normal melanosome morphogenesis and eumelanin deposition. A nonsense mutation that leads to a marked truncation of the deduced protein was found in PMEL of the mutant. The gene expression level of PMEL decreased substantially in the mutant. Genotypes at the site of the nonsense mutation were fully concordant with plumage colour phenotypes in 196 F2 offspring. The nonsense mutation was not found in several quail strains with non-yellowish plumage. Thus, the yellowish plumage may be caused by the reduced eumelanin content in feathers because of the loss of PMEL function.

Chicken egg white ovotransferrin (OTF) has been reported to exist in three electrophoretic variants (OTFA, OTFB and OTFC). In this report, we identified a causal polymorphism between the OTFB and OTFC variants in Japanese and Taiwanese native chickens and compared the antibacterial activity between these two variants. The cDNA sequence analyses from Satsumadori oviducts revealed that three non-synonymous SNPs T1809G (Ser52Ala), A2258G (Ile96Val) and G7823A (Asp500Asn) corresponded to the OTF electrophoretic phenotypes. Of the three SNPs, the G7823A mutation perfectly corresponded to the electrophoretic phenotypes OTFB (G/G, Asp500Asp), OTFB/C (G/A, Asp500Asn) and OTFC (A/A, Asn500Asn) in three chicken populations. The variants OTFB and OTFC exhibited similar antibacterial potency against Gram-positive and Gram-negative bacteria. This study provides, for the first time, molecular information on polymorphism of OTFB and OTFC variants of chicken ovotransferrin and its effect on the antimicrobial activity of the respective variants.

Phenalenyl, zethrene, and extended zethrene molecules were employed to study the change in electronic properties of graphene surfaces upon exposure to oxygen atmosphere. These hydrocarbons are the ...

The Kurokashiwa breed of native Japanese chickens is primarily reared in Shimane and Yamaguchi Prefectures, Japan. To reveal the genetic diversity and differentiation among the Kurokashiwa populations in both prefectures, 29 microsatellites were analyzed. To prepare the microsatellites, 54, 55, and 24 blood samples were collected from the Kurokashiwa populations of five fancy breeders in Shimane Prefecture, three fancy breeders in Yamaguchi Prefecture, and the Livestock Technology Research Department, Yamaguchi Prefectural Agriculture and Forestry General Technology Center (LTRDY), respectively. The corrected number of alleles (allelic richness: AR) was considerably low (1.63-2.48) through all Shimane and Yamaguchi populations. However, inbreeding coefficient (F-IS) values were not significant in the entire Shimane and Yamaguchi ornamental populations as well as the LTRDY population. In addition, there was no statistically significant difference in expected heterozygosity (H-E) through all populations in Shimane and Yamaguchi Prefectures, although the H-E varied from 0.237 to 0.445 depending on populations. The topology of the phylogenetic tree, pairwise F-ST values, and STRUCTURE analysis indicated that Kurokashiwa populations were genetically separated between Shimane and Yamaguchi Prefectures. On the other hand, there was no genetic differentiation among ornamental populations within each prefecture, which seemed to have resulted from adequate random matings by exchanging individuals among fancy breeders within each conservation community, along with a proper random mating within LTRDY. The present study proposes a conservation strategy suggesting that the Kurokashiwa populations of Shimane and Yamaguchi Prefectures should be maintained separately with continuous exchange of birds within each conservation community to preserve genetic diversity. Also, the LTRDY population should be independently kept by avoiding matings with ornamental bird populations because it is a well established closed colony with a uniform genetic constitution.

1. The E3 ubiquitin protein ligase 1 (WWP1) gene, the mutation of which causes muscular dystrophy in chickens, is expressed not only in the pectoral muscle, but also in a number of tissues such as the kidney. Therefore, this study examined some parameters related to kidney function in muscular dystrophic (MD) chickens. 2. Plasma osmolality, Na+ and K+ concentrations, aldosterone levels, and the expression of aquaporin (AQP) 2, AQP3, and α subunits of the amiloride-sensitive epithelial sodium channel (αENaC) were analysed in the kidneys of 5-week-old MD chickens and White Leghorn (WL) chickens under physiological conditions or after one day of water deprivation. 3. Plasma osmolality, Na+ concentrations, and plasma aldosterone levels were significantly higher in MD chickens than in WL chickens. αENaC mRNA expression levels were lower in MD chickens than in WL chickens. AQP2 and AQP3 mRNA expression levels were similar in the two strains of chickens. 4. Plasma osmolality correlated with aldosterone levels and AQP2 and αENaC mRNA levels in WL chickens. In MD chickens, plasma osmolality correlated with AQP2 mRNA levels, but not with plasma aldosterone or αENaC mRNA levels. 5. These results suggest that neither water reabsorption nor the expression of AQP2 and AQP3 is impaired in MD chickens and that a WWP1 gene mutation may or may not directly induce an abnormality in Na+-reabsorption in the kidneys of MD chickens, potentially through αENaC.

It is assumed that Red Junglefowl (Gallus gallus) is one of the main ancestors of domestic chickens (Gallus gallus domesticus). Differences in microsatellite polymorphisms between Red Junglefowl and modern commercial chickens, which are used for egg and meat production, have not been fully reported. A total of 361 individuals from 1 Red Junglefowl population that has been maintained as a closed flock, 5 final cross-bred commercial layer populations (white-, tinted-, and brown-egg layers), and 2 final cross-bred commercial broiler populations were genotyped for 40 autosomal microsatellite loci. We compared microsatellite variations in Red Junglefowl with those in a commercial chicken gene pool. The contribution of each population to the genetic diversity was also estimated based on the molecular coancestry. In total, 302 distinct alleles were detected in 1 Red Junglefowl and 7 commercial chicken populations, of which 31 alleles (10.3%) were unique to Red Junglefowl, most of which occurred at a high frequency. The genetic differentiation between Red Junglefowl and commercial chickens (pairwise FST) ranged from 0.32 to 0.47. According to the neighbor-joining tree based on the modified Cavalli-Sforza chord distances and the Bayesian clustering analysis, Red Junglefowl was genetically distant from the commercial chicken gene pool tested. In all of the populations analyzed, Red Junglefowl made the highest contribution to genetic diversity. These results suggest that Red Junglefowl has a distinct distribution of microsatellite alleles and that there is a high level of genetic divergence between Red Junglefowl and commercial chickens.

We experienced two cases of unrelated Japanese children with bacille Calmette-Guérin (BCG) multiple osteomyelitis with partial interferon (IFN)-γ receptor 1 (IFNGR1) deficiency. Heterozygous small deletions with frame shift (811 del4 and 818 del4) were detected, which were consistent with the diagnosis of partial dominant IFNGR1 deficiency. Case 1: a 2-year-old boy visited us because of limb and neck pain. He had been vaccinated with BCG at 17 months of age. Multiple destructive lesions were observed in the skull, ribs, femur, and vertebral bones. Mycobacterium bovis (BCG Tokyo 172 strain by RFLP technique) was detected in the bone specimen. The BCG multiple osteomyelitis was treated successfully without recurrence. Case 2: an 18-month-old girl developed multiple osteomyelitis 9 months after BCG inoculation. Radiologic images showed multiple osteolytic lesions in the skull, ribs, femur, and vertebrae. M. bovis (BCG Tokyo 172 strain) was detected in the cultures from a bone biopsy. Her clinical course showed recurrent osteomyelitis and lymphadenitis with no pulmonary involvement. The effects of high-dose antimycobacterial drugs and IFN-γ administration were transient, and complete remission has since been achieved by combination antimycobacterial therapy, including levofloxacin. Partial dominant IFNGR1 deficiency is a rare disorder, but it should be considered when a patient presents with multiple osteomyelitis after BCG vaccination. The cases that are resistant to conventional regimens require additional second-line antituberculous drugs, such as levofloxacin.

Interspecific hybrid incompatibility, including inviability and sterility, is important in speciation; however, its genetic basis remains largely unknown in vertebrates. Crosses between male chickens and female Japanese quails using artificial insemination can generate intergeneric hybrids; however, the hatching rate is low and hatched hybrids are only sterile males. Hybrid development is arrested frequently during the early embryonic stages and the sex ratio of living embryos is male-biased. However, the development and sex ratio of hybrid embryos have not been comprehensively analyzed. In the present study, we observed delayed embryonic development of chicken-quail hybrids during the early stage, compared with that of chickens and quails. The survival rate of hybrids decreased markedly during the blastoderm-to-pre-circulation stage and then decreased gradually through the subsequent stages. Hybrid females were observed at more than 10 d of incubation; however, the sex ratio of hybrids became male-biased from 10 d of incubation. Severely malformed embryos were observed frequently in hybrids. These results suggest that developmental arrest occurs at various stages in hybrid embryos, including a sexually non-biased arrest during the early stage and a female-biased arrest during the late stage. We discuss the genetic basis for hybrid inviability and its sex bias.

A number of crested chicken strains, such as the Polish chicken, exhibit a bony protuberance in the anterodorsal region of their skulls. The shape of their brain shows the anatomical peculiarity that is characterized by the upthrusting of cerebral hemispheres, called "cerebral hernia." Some early works suggested that this phenotype may be caused by a genetic factor and any modifiers influencing the development of brain and/or skull. However, the causative gene and its formation mechanism are still unclear. The present study is aimed to analyze the inheritance and ontogenic process of cerebral hernia in the crested Polish chicken. Firstly, we constructed the resource family with the Polish chicken and PNP inbred strain. Genetic analysis of this family revealed that cerebral hernia is controlled by a single autosomal recessive gene and is closely associated with crest formation. Furthermore, our morphological analysis of brain structures in the progenies suggested that the significant enlargement of brain cavity at later stages of embryos, particularly after 15 days of incubation, may be the main cause of cerebral hernia.

Recently, Hirao and coworkers have reported the synthesis of cyclopentadienyl iron complexes of sumanene. From their experimental works, it was found that the concave-bound cyclopentadienyl iron complex of sumanene was only obtained in the crystal prepared. Conversely, theoretical calculations showed the convex-bound cyclopentadienyl iron complex of sumanene was relatively stable than the concave-bound one. Thus, there is a confliction between the experimental and theoretical research results. To solve this problem, the density functional theory calculations were carried out for the several types of cyclopentadienyl iron complexes with/without AlCl ion. These calculations suggest that AlCl ion plays an important role for the stabilization of the concave-bound cyclopentadienyl iron complex of sumanene. © 2012 Wiley Periodicals, Inc.

During early development in vertebrates, pluripotent cells are generated from the neural crest and migrate according to their presumptive fate. In birds and mammals, one of the progeny cells, melanoblasts, generally migrate through a dorsolateral route of the trunk region and differentiate to melanocytes. However, Silky is an exceptional chicken in which numerous melanoblasts travel via a ventral pathway and disperse into internal organs. Finally, these ectopic melanocytes induce heavy dermal and visceral melanization known as Fibromelanosis (Fm). To identify the genetic basis of this phenotype, we confirmed the mode of inheritance of Fm as autosomal dominant and then performed linkage analysis with microsatellite markers and sequence-tagged site markers. Using 85 backcross progeny from crossing Black Minorca chickens (BM-C) with F1 individuals between White Silky (WS) and BM-C Fm was located on 10.2–11.7 Mb of chicken chromosome 20. In addition, we noticed a DNA marker that all Silky chickens and the F1 individuals showed heterozygous genotyping patterns, suggesting gene duplication in the Fm region. By quantitative real-time PCR assay, Silky line-specific gene duplication was detected as an ∼130-kb interval. It contained five genes including endothelin 3 (EDN3), which encoded a potent mitogen for melanoblasts/melanocytes. EDN3 with another three of these duplicated genes in Silky chickens expressed almost twofold of those in BM-C. Present results strongly suggest that the increase of the expression levels resulting from the gene duplication in the Fm region is the trigger of hypermelanization in internal organs of Silky chickens.

As IgM antibody measurement by enzyme-linked immunosorbent assay (ELISA) has become possible for the serological diagnosis of Chlamydophila pneumoniae (C. pn) infection, the HITAZYME-ELISA method has become widely employed in Japan. However, in children, when the diagnostic criterion of primary infection is set at ID ≥1.1, the positive rate is higher than expected, and the potential for inaccurate reflection of the prevalence has been raised. In this study, we performed ROC analysis involving 136 pediatric patients with acute airway symptoms (0-14 years of age), considering a 32-fold or higher micro-immunofluorescence IgM antibody titer against C. pn as positive. Setting the cut-off value for ELISA C. pn IgM antibody ID at 2.0, the specificity was 100%, with no false positivity. The maximum (sensitivity + specificity)/2 was obtained when the cut-off value was set at 1.5. Therefore, IgM ID ≥2.0 was regarded as definitely positive and an IgM ID between 1.5 and 2.0 was regarded as indeterminate as diagnostic criteria for the primary infection. When the prevalence was investigated in 3,108 children (0-15 years of age) with airway symptoms based on these criteria, 542 cases (17.4%) were positive, and the median duration of IgM antibody positivity was five months. Long-term positivity (ten cases) for more than 12 months and recurrent positivity (eight cases) were also observed, but it may be appropriate to set a new criterion of IgM antibody ID ≥2.0 for the diagnosis of primary Chlamydophila pneumoniae infection in children.

Japanese quail egg white lysozyme possesses two alleles (S and F). In our previous study, the F phenotype was more effective against bacteria than the S phenotype, whereas the female-derived S phenotype led to improved hatchability. In this study, we analyzed gene frequency of egg white lysozyme using Acid-PAGE and mismatch PCR-RFLP in 22 Japanese quail populations in Japan and compared with five populations in INRA, France. Genotyping of the lysozyme locus in 22 Japanese quail populations in Japan revealed that this locus is polymorphic in all populations except Nagoya University and Kagoshima University (LL and SS) populations. The S allele was a major allele with frequency from 0.69 to 1.00. In five French populations, it was polymorphic in all populations except AA population. The S allele was a major allele with frequency from 0.69 to 1.00. From these results, the lysozyme locus was polymorphic in the Japanese quails and the S allele was a major allele. The S allele was fixed in several populations which have been maintained as closed and small population.

The effective exchange integrals ( J (HB)) of the Heisenberg spin model have been evaluated by using the ab initio MO and based on Hartree–Fock (HF) and density functional theory (DFT) for organic magnetic metals, the (EDT-TTFVO) 2 FeBr 4 crystal based on the X-ray crystallographic structures at 113 K. In order to study the magnetic properties, we proposed some of the pairs, where the direct ( d – d ) and indirect ( d –π– d ) magnetic couplings between Fe(III) d -spins ( S = 5/2) without/with π-dimer spins ( S = 1/2) were calculated, respectively. The effective exchange integrals were evaluated by using UB3LYP method, and principal J values were 0.5, −0.1 and 0.4 K. From these results, it is found that there were three dimensional spin arrangements of Fe(III) d -spins. The Quantum Monte Carlo (QMC) simulations had been carried out with our calculated J values to evaluate the magnetic susceptibility for this molecular crystal, reproducing the experimental tendency.

Intermolecular interactions in nano-porous crystals were studied theoretically as related to adsorption properties. The crystal [Rh 2 bza 4 pyz] n , which had been reported by Mori, Takamizawa and their co-workers in detailed, was employed in this study. As adsorbed magnetic radicals, a oxygen molecule (O 2 ; S = 1) was treated. Adsorption energies Δ E ads are defined for quantitative discussion for host–guest interaction. From the calculated results by our MO calculations, weak attractive interactions between adsorbed O 2 and surrounding four ph-groups (delegated to benzene) existed. In addition, stable structure of O 2 molecules over one benzene molecule was studied. From the results, parallel structure is more stable than perpendicular structure. Physisorption phenomena in intermediate distance region ( R = 3.4 A) contributes to energy stabilization.