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Ultrasound can provide real-time imagery without the risk of radiation exposure, and it is widely available at a relatively low cost. It can provide updated three-dimensional information that can improve the physician’s spatial awareness during a brachytherapy procedure for cervical cancer. There is mounting evidence demonstrating the numerous benefits of ultrasound-guided brachytherapy in the published literature. This evidence supports its routine use to improve the safety and the effectiveness of cervical brachytherapy. In this report we will review various methods in which ultrasound imaging has been used during cervical brachytherapy. We also include a description of our own institutional approach to ultrasound-guided cervical implementation that has been in use for all cervical brachytherapy procedures over the past two decades.

Genetic parameters for carcass grading traits, image analysis traits, and monounsaturated fatty acid (MUFA) percentages were estimated in 29,942 Japanese Black cattle from Hyogo Prefecture. The analyzed traits included five carcass grading traits, two image analysis traits, fat area ratio and fineness index, and two MUFA traits, one measured in intermuscular fat using near-infrared spectroscopy (NIRS) and the other in intramuscular fat using gas chromatography (GC). The heritability estimates of image analysis traits and MUFA were moderate to high, ranging from 0.395 to 0.740, and it was considered that they could be improved simultaneously with carcass grading traits because no severe genetic antagonism was observed. Although the heritability of the NIRS-based intermuscular MUFA was slightly lower than that of the GC-based intramuscular MUFA, the genetic correlation between the two methods was as high as 0.804. These results indicate that the NIRS method can be used as an alternative evaluation procedure to predict MUFA in intramuscular fat in the longissimus muscle.

In our previous study, we used genome resequencing to detect all candidate polymorphisms within a quantitative trait loci (QTL) region for beef marbling reported previously at 10–30 Mbp on bovine chromosome 7, and we selected 6044 polymorphisms as candidate quantitative trait nucleotides (QTNs). In the present study, we aimed to identify quantitative trait genes (QTGs) and QTNs in this QTL region by verifying the effect of SNPs on beef marbling in two Japanese Black cattle populations using a Dynamic Array integrated fluidic circuit. In total, 96 selected SNPs were genotyped in 441 and 529 animals in Hyogo and Miyazaki cattle populations, respectively. The most significant p-values were detected in a SNP in a splice region of ALDH7A1 (SNP93_ALDH7A1; p = 3.46 × 10−5) in Hyogo cattle and a missense polymorphism of intercellular adhesion molecule-1 (ICAM1) (SNP37_ICAM1; p = 3.33 × 10−4) in Miyazaki cattle. Interestingly, SNP93_ALDH7A1 was not significant (p = 0.459) in Miyazaki cattle, and SNP37_ICAM1 showed a weakly significant association (p = 0.043) in Hyogo cattle. Thus, each population would likely have different QTGs and QTNs for beef marbling in the QTL region. In the Hyogo population, it was not possible to determine the accurate range of the linkage disequilibrium (LD) block in LD block analysis because of a strong LD structure throughout the assessed region. In Miyazaki cattle, however, an LD block containing SNP37_ICAM1 had a range of 15.8–16.1 Mbp, suggesting that QTNs would be located within this region. The functions of 19 genes in the LD block were investigated. ICAM1 is known to play an important role in adipocyte differentiation; given this function and the effect of amino acid substitution, SNP37_ICAM1 was identified as a promising candidate QTN for beef marbling. Further research on the effect of SNP37_ICAM1 on adipocyte differentiation is expected to provide insights into the mechanism underlying beef marbling formation.

American foulbrood (AFB) and European foulbrood (EFB) caused by Paenibacillus larvae and Melissococcus plutonius, respectively, are major bacterial infections of honey bees. Although macrolides (mirosamicin [MRM] and tylosin) have been used to prevent AFB in Japan, macrolide-resistant P. larvae have yet to be found. In this study, we revealed that both MRM-resistant and -susceptible strains exist in Japanese M. plutonius and that a methyltransferase gene (rlmA II ) was disrupted exclusively in MRM-susceptible strains due to a single-nucleotide insertion. The M. plutonius RlmAII modified G748 of 23S rRNA, and the deletion of rlmA II resulted in increased susceptibility to MRM and the loss of modification at G748, suggesting that methylation at G748 by RlmAII confers MRM resistance in M. plutonius. The single-nucleotide mutation in MRM-susceptible strains was easily repaired by spontaneous deletion of the inserted nucleotide; however, intact rlmA II was only found in Japanese M. plutonius and not in a Paraguayan strain tested or any of the whole-genome-sequenced European strains. MRM has been used in apiculture only in Japan. Although M. plutonius is not the target of this drug, the use of MRM as a prophylactic drug for AFB may have influenced the antibiotic susceptibility of the causative agent of EFB.

Paenibacillus larvae is the causative agent of American foulbrood (AFB), the most destructive disease of the honey bee brood. In this study, we investigated the population structure and antimicrobial susceptibility of Japanese P. larvae using 100 isolates isolated between 1993 and 2017 in 17 prefectures. Using repetitive-element PCR and multilocus sequence typing, isolates from diverse origins were classified into six genotypes, including the novel genotype ERIC II-ST24. Among these genotypes, ERIC I-ST15 is the most common in Japan, while ERIC II-ST10 isolates were found to be increasing during the 2010s. Regardless of genotype or origin, all isolates were susceptible to the major antimicrobials used in the control of AFB, including mirosamicin and tylosin, which were approved for the prevention of AFB in Japan in 1999 and 2017 respectively. Despite nearly 20 years of use, mirosamicin is still effective against Japanese P. larvae in vitro; however, the development of AFB in honey bee colonies may not always be suppressed by this drug. The case information collected in this study provides insight into the conditions under which prophylactic medicine may not exert sufficient preventive effects in vivo.

The objective of this study was to identify genomic regions associated with fat-related traits using a Japanese Black cattle population in Hyogo. From 1836 animals, those with high or low values were selected on the basis of corrected phenotype and then pooled into high and low groups (n = 100 each), respectively. DNA pool-based genome-wide association study (GWAS) was performed using Illumina BovineSNP50 BeadChip v2 with three replicate assays for each pooled sample. GWAS detected that two single nucleotide polymorphisms (SNPs) on BTA7 (ARS-BFGL-NGS-35463 and Hapmap23838-BTA-163815) and one SNP on BTA12 (ARS-BFGL-NGS-2915) significantly affected fat percentage (FAR). The significance of ARS-BFGL-NGS-35463 on BTA7 was confirmed by individual genotyping in all pooled samples. Moreover, association analysis between SNP and FAR in 803 Japanese Black cattle revealed a significant effect of SNP on FAR. Thus, further investigation of these regions is required to identify FAR-associated genes and mutations, which can lead to the development of DNA markers for marker-assisted selection for the genetic improvement of beef quality.

In our previous study, we performed genome-wide association study (GWAS) to identify the genomic region associated with Fat area ratio to rib eye area (FAR) and detected a candidate in BTA7 at 10-30 Mbp. The present study aims to comprehensively detect all polymorphisms in the candidate region using whole-genome resequencing data. Based on whole-genome resequencing of eight animals, we detected 127,090 polymorphisms within the region. Of these, 31,945 were located within the genes. We further narrowed the polymorphisms to 6,044 with more than five allele differences between the high and low FAR groups that were located within 179 genes. We subsequently investigated the functions of these genes and selected 170 polymorphisms in eight genes as possible candidate polymorphisms. We focused on SLC27A6 K81M as a putative candidate polymorphism. We genotyped the SNP in a Japanese Black population (n = 904) to investigate the effect on FAR. Analysis of variance revealed that SLC27A6 K81M had a lower p-value (p = .0009) than the most significant SNP in GWAS (p = .0049). Although only SLC27A6 K81M was verified in the present study, subsequent verification of the remaining candidate genes and polymorphisms could lead to the identification of genes and polymorphisms responsible for FAR.

American foulbrood (AFB) and European foulbrood (EFB) caused by Paenibacillus larvae and Melissococcus plutonius, respectively, are major bacterial infections of honey bees. Although macrolides (mirosamicin [MRM] and tylosin) have been used to prevent AFB in Japan, macrolide-resistant P. larvae have yet to be found. In this study, we revealed that both MRM-resistant and -susceptible strains exist in Japanese M. plutonius and that a methyltransferase gene (rlmA

Marker-assisted selection (MAS) is expected to accelerate the genetic improvement of Japanese Black cattle. However, verification of the effects of the genes for MAS in different subpopulations is required prior to the application of MAS. In this study, we investigated the allelic frequencies and genotypic effects for carcass traits of six genes, which can be used in MAS, in eight local subpopulations. These genes are SCD, FASN and SREBP1, which are associated with the fatty acid composition of meat, and NCAPG, MC1R and F11, which are associated with carcass weight, coat color and blood coagulation abnormality, respectively. The frequencies of desirable alleles of SCD and FASN were relatively high and that of NCAPG was relatively low, and NCAPG was significantly associated with several carcass traits, including carcass weight. The proportions of genotypic variance explained by NCAPG to phenotypic variance were 4.83 for carcass weight. We thus confirmed that NCAPG is a useful marker for selection of carcass traits in these subpopulations. In addition, we found that the desirable alleles of six genes showed no negative effects on carcass traits. Therefore, selection using these genes to improve target traits should not have negative impacts on carcass traits.

Campylobacter-induced diarrhea is increasingly recognized worldwide. However, little information is available regarding the Campylobacter strains associated with diarrheal patients in Thailand. In this study, we attempted to isolate Campylobacter strains from diarrheal patients in Thailand and to characterize the species using a cytolethal distending toxin (cdt) gene-based C. jejuni, C. coli, and C. fetus-specific multiplex PCR assay. Campylobacter species were also confirmed using 16S rRNA gene sequencing and hipO gene detection. From 2,500 diarrheal stool specimens, 76 Campylobacter-like organisms were isolated and identified via conventional culture methods. Among these 76 organisms, 73 were identified as Campylobacter species (43 C. jejuni, 29 C. coli, and 1 C. fetus) via multiplex PCR, whereas 3 remained unidentified. Two Campylobacter-like organisms yielded 2 amplicons corresponding to cdt genes from C. jejuni and C. coli. Subsequently, C. jejuni and C. coli were reisolated from each sample. The third isolate was identified as C. hyointestinalis via 16S rRNA gene sequencing. To our knowledge, this is the first report on the isolation of C. hyointestinalis from a diarrheal patient in Thailand. These data indicate that C. jejuni (58%) and C. coli (40%) are prevalent among diarrheal patients in Thailand.

Paenibacillus larvae is the causative agent of American foulbrood (AFB), the most destructive disease of the honey bee brood. In this study, we investigated the population structure and antimicrobial susceptibility of Japanese P. larvae using 100 isolates isolated between 1993 and 2017 in 17 prefectures. Using repetitive-element PCR and multilocus sequence typing, isolates from diverse origins were classified into six genotypes, including the novel genotype ERIC II-ST24. Among these genotypes, ERIC I-ST15 is the most common in Japan, while ERIC II-ST10 isolates were found to be increasing during the 2010s. Regardless of genotype or origin, all isolates were susceptible to the major antimicrobials used in the control of AFB, including mirosamicin and tylosin, which were approved for the prevention of AFB in Japan in 1999 and 2017 respectively. Despite nearly 20 years of use, mirosamicin is still effective against Japanese P. larvae in vitro; however, the development of AFB in honey bee colonies may not always be suppressed by this drug. The case information collected in this study provides insight into the conditions under which prophylactic medicine may not exert sufficient preventive effects in vivo.

Fatty acid composition is an important indicator of beef quality. The objective of this study was to search the potential candidate region for fatty acid composition. We performed pool-based genome-wide association studies (GWAS) for oleic acid percentage (C18:1) in a Japanese Black cattle population from the Hyogo prefecture. GWAS analysis revealed two novel candidate regions on BTA9 and BTA14. The most significant single nucleotide polymorphisms (SNPs) in each region were genotyped in a population (n = 899) to verify their effect on C18:1. Statistical analysis revealed that both SNPs were significantly associated with C18:1 (p = .0080 and .0003), validating the quantitative trait loci (QTLs) detected in GWAS. We subsequently selected VNN1 and LYPLA1 genes as candidate genes from each region on BTA9 and BTA14, respectively. We sequenced full-length coding sequence (CDS) of these genes in eight individuals and identified a nonsynonymous SNP T66M on VNN1 gene as a putative candidate polymorphism. The polymorphism was also significantly associated with C18:1, but the p value (p = .0162) was higher than the most significant SNP on BTA9, suggesting that it would not be responsible for the QTL. Although further investigation will be needed to determine the responsible gene and polymorphism, our findings would contribute to development of selective markers for fatty acid composition in the Japanese Black cattle of Hyogo.

Background and objective The yield of biopsy performed during bronchoscopy is reduced if the lesion is smaller than 30 mm. We evaluated the performance of a new diagnostic technique combining endobronchial ultrasonography with a guide sheath (EBUS-GS) and a virtual bronchoscopic navigation system, LungPoint (Broncus Technologies, Inc., Mountain View, CA, USA), for the diagnosis of small (≤30 mm) peripheral pulmonary lesions (PPL). Methods Between May 2011 and December 2011, we recruited 68 consecutive patients presenting with a PPL 30 mm or less in diameter determined by chest computed tomography. We used the LungPoint system before bronchoscopy to identify the bronchus into which the bronchoscope should be advanced. We used a thin bronchoscope. EBUS-GS was performed using an endoscope ultrasonography system equipped with a 20-MHz mechanical radial-type probe. We used a guide sheath with an external diameter of 1.95 mm, thin forceps and brushing. Results The diagnostic yield of the 68 PPL was 77.9%; it was 83.7% and 68.0% for the malignant and benign lesions, respectively. Notably, three cases were diagnosed by transbronchial needle-aspiration cytology alone. Univariate and multivariate analyses showed that the EBUS probe localization was the most significant contributor to successful diagnosis (diagnostic yield: within vs adjacent to the lesion = 92.1% vs 60.0%, respectively; P = 0.004 and P = 0.003, respectively). Conclusions The combination of EBUS-GS and LungPoint was useful for diagnosing small PPL.

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.

スギ製材工程で大量に発生する樹皮の有効利用を図るため，スギ樹皮のアンモニア吸着能に着目した鶏ふん堆肥化過程の臭気低減効果について検討した。粉砕処理したスギ樹皮を620ppmに調製したバッグ内のアンモニアガスと5分間接触させたところ99.5％のアンモニアが除去された。鶏ふん堆積物の表面をスギオガクズまたはスギ樹皮で被覆し，堆肥化過程で発生するアンモニアの吸着を試みたところ，スギオガクズは1g当たり11mgのアンモニアを吸着したのに対し，スギ樹皮では33mgに達した。堆肥化後，スギ樹皮中の窒素の形態を調査したところ，水溶性窒素が22％，イオン交換性窒素が23％，有機態窒素47％であり，高い陽イオン交換能に伴うイオン結合だけでなく，共有結合による除去も示唆された。スギ樹皮は，アンモニアガス発生源の表面を被覆するだけで発生量を抑制でき，安価で簡便な臭気対策資材としての活用が期待される。

We have recently developed a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for identifying Campylobacter jejuni, C. coli and C. fetus. In the present study, the applicability of this assay was evaluated with 34 Campylobacter-like organisms isolated from poultry in Thailand for species identification and was compared with other assays including API Campy, 16S rRNA gene sequence, and hippuricase (hipO) gene detection. Of the 34 strains analyzed, 20, 10 and 1 were identified as C. jejuni, C. coli, and Arcobacter cryaerophilus, respectively, and 3 could not be identified by API Campy. However, 16S rRNA gene analysis, showed that all 34 strains are C. jejuni/coli. To discriminate between these 2 species, the hipO gene, which is specifically present in C. jejuni, was examined by PCR and was detected in 20 strains, which were identified as C. jejuni by API Campy but not in the remaining 14 strains. Collective results indicated that 20 strains were C. jejuni whereas the 14 strains were C. coli. When the cdt gene-based multiplex PCR was employed, however, 19, 20 and 19 strains were identified as C. jejuni while 13, 14 and 13 were identified as C. coli by the cdtA, cdtB and cdtC gene-based multiplex PCR, respectively. Pulsed-field gel electrophoresis revealed that C. jejuni and C. coli strains analyzed are genetically diverse. Taken together, these data suggest that the cdt gene-based multiplex PCR, particularly cdtB gene-based multiplex PCR, is a simple, rapid and reliable method for identifying the species of Campylobacter strains.

We have cloned a gene which is specifically expressed at the stage of sexual maturation in the rat testis by means of differential display, and have named it spermatogenesis-related factor-2 (SRF-2). Testicular expression was first detected at 5 weeks of age, and its level of the expression increased up to 7 weeks, and was maintained even at 63 weeks. Its cDNA was 2,789 bp in length and encoded an open reading frame of 718 amino acids. This gene was mainly expressed in the spermatocyte, judging from the result of in situ hybridization. The hypothetical gene product had a motif highly homologous with RabGAP/TBC protein. Taken together, this gene is considered to have some important functions for meiosis. The gene expression was significantly decreased by treatment with TCDD, a candidate endocrine disruptor, when administered to male rats of the nursling period. Body weight and testis weight were decreased by the treatment, but even then the sperm concentration in cauda epididymis was not changed significantly. SRF-2 gene may be a promising biomarker to construct a detection system of uncertain endocrine disruptors.

Zirconium-mediated diastereoselective alkene-carbonyl coupling reactions of N-alkenylcarbamate derivatives are reported. The present alkene-carbonyl coupling reactions using chiral tert-butyl 3-butenylcarbamates having a substituent at the homoallylic position proceeded in a highly diastereoselective manner to give α-methyl-γ-substituted γ-aminobutyric acid (GABA) derivatives. Iodination of the intermediary zirconium species gave the u-iodo-methylated γ-aminobutyrates, which were, in turn, converted to the 5-substituted pyrrolidine-3-carboxylate having cis stereochemistry.

Chlorophyll (Chl) a', the C132-epimer of Chl a, is a constituent of the primary electron donor (P700) of Photosystem (PS) I of a thermophilic cyanobacterium Synechococcus (Thermosynechococcus) elongatus, as was recently demonstrated by X-ray crystallography. To determine whether PS I of oxygenic photosynthetic organisms universally contains one molecule of Chl a', pigment compositions of thylakoid membranes and PS I complexes isolated from the cyanobacteria T. elongatus and Synechocystis sp. PCC 6803, the green alga Chlamydomonas reinhardtii, and the green plant spinach, were examined by simultaneous detection of phylloquinone (the secondary electron acceptor of PS I) and Chl a' by reversed-phase HPLC. The results were compared with the Chl a/P700 ratio determined spectrophotometrically. The Chl a'/PS I ratios of thylakoid membranes and PS I were about 1 for all the organisms examined, and one Chl a' molecule was found in PS I even after most of the peripheral subunits were removed. Chl a' showed a characteristic extraction behaviour significantly different from the bulk Chl a in acetone/methanol extraction upon varying the mixing ratio. These findings confirm that a single Chl a' molecule in P700 is the universal feature of PS I of the Chl a-based oxygenic photosynthetic organisms.

Campylobacter-induced diarrhea is increasingly recognized worldwide. However, little information is available regarding the Campylobacter strains associated with diarrheal patients in Thailand. In this study, we attempted to isolate Campylobacter strains from diarrheal patients in Thailand and to characterize the species using a cytolethal distending toxin (cdt) gene-based C. jejuni, C. coli, and C. fetus-specific multiplex PCR assay. Campylobacter species were also confirmed using 16S rRNA gene sequencing and hipO gene detection. From 2,500 diarrheal stool specimens, 76 Campylobacter-like organisms were isolated and identified via conventional culture methods. Among these 76 organisms, 73 were identified as Campylobacter species (43 C. jejuni, 29 C. coli, and 1 C. fetus) via multiplex PCR, whereas 3 remained unidentified. Two Campylobacter-like organisms yielded 2 amplicons corresponding to cdt genes from C. jejuni and C. coli. Subsequently, C. jejuni and C. coli were reisolated from each sample. The third isolate was identified as C. hyointestinalis via 16S rRNA gene sequencing. To our knowledge, this is the first report on the isolation of C. hyointestinalis from a diarrheal patient in Thailand. These data indicate that C. jejuni (58%) and C. coli (40%) are prevalent among diarrheal patients in Thailand.

Marker-assisted selection (MAS) is expected to accelerate the genetic improvement of Japanese Black cattle. However, verification of the effects of the genes for MAS in different subpopulations is required prior to the application of MAS. In this study, we investigated the allelic frequencies and genotypic effects for carcass traits of six genes, which can be used in MAS, in eight local subpopulations. These genes are SCD, FASN and SREBP1, which are associated with the fatty acid composition of meat, and NCAPG, MC1R and F11, which are associated with carcass weight, coat color and blood coagulation abnormality, respectively. The frequencies of desirable alleles of SCD and FASN were relatively high and that of NCAPG was relatively low, and NCAPG was significantly associated with several carcass traits, including carcass weight. The proportions of genotypic variance explained by NCAPG to phenotypic variance were 4.83 for carcass weight. We thus confirmed that NCAPG is a useful marker for selection of carcass traits in these subpopulations. In addition, we found that the desirable alleles of six genes showed no negative effects on carcass traits. Therefore, selection using these genes to improve target traits should not have negative impacts on carcass traits.

A series of nucleocapsid protein (NP)-deleted genes of the Onderstepoort strain was constructed in order to locate antigenic regions of the NP of canine distemper virus. The expression of proteins from 5'-deleted NP genes was examined in COS-7 cells by indirect immunofluorescence assay using three monoclonal antibodies (MAbs), c-5, f-5 and h-6, and a rabbit serum against NP. These MAbs reacted with two regions of NP. Amino acid residues from 1 to 80, and 337-358, were necessary and sufficient for formation of the epitopes identified by MAbs f-5 and h-6, and c-5, respectively. The proteins translated from intact or 3'-deleted genes were found to be localized in the nuclei of COS-7 cells, whereas the proteins from the 5'-deleted genes were mainly detected in the cytoplasm. These results suggested that 80 amino acid residues at the N-terminus are required for transportation of NP into the nucleus.

Cap structures capable of sustaining infinite growth by ingestion of C2 fragments to give a carbon nanotube were screened systematically. For this purpose a total of 23,296 IPR cap structures having characteristic tubule vectors of (5∼20, 0∼10) were generated by the use of net algorithm. Eighteen recurring patterns were found which involve 50 cap structures having tubule vectors (6∼11, 5).

We analyzed the molecular properties of the nucleocapsid protein (NP) of canine distemper viruses (CDV), isolated between 1992 and 1995 in Japan. Four CDV field isolates (Yanaka, Ueno, Hamamatsu, and Adachi strains) obtained were antigenically identical. Sequence analysis of entire region of the NP gene of a field isolate, the Yanaka strain, revealed that the NP gene contained 1683 nucleotides and was 93.2% homologous with a laboratory strain, the Onderstepoort strain. The deduced amino acid sequence contained 523 amino acids and was 95.2 and 99% homologous with those of the Onderstepoort and a virulent strain, A75 17 strain, respectively. Since most of the diversities in amino acid sequence occurred in two domains, at the N′- and the C′- termini, we further sequenced 3′-terminal regions of the remaining three field isolates. Based on the sequences, the new CDV isolates had one cluster that distinguished them from the laboratory strain.

Analysis of the molecular properties of fusion (F) proteins of field isolates of canine distemper virus (CDV) by immunoprecipitation analysis revealed an identical molecular mass of F protein of 3 field isolates as well as the Onderstepoort laboratory strain. Sequencing showed that the F gene of a field isolate (the Yanaka strain) shared 90.1% and 95.7% identities with the Onderstepoort strain at nucleotide and amino acid levels, respectively. All of the 13 cysteine residues and 4 potential asparagine-linked glycosylation sites were completely conserved amongst these strains. These results indicate that the F proteins is much less heterogeneous than that observed in the hemagglutinin proteins of CDV.

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.

The yield of biopsy performed during bronchoscopy is reduced if the lesion is smaller than 30 mm. We evaluated the performance of a new diagnostic technique combining endobronchial ultrasonography with a guide sheath (EBUS-GS) and a virtual bronchoscopic navigation system, LungPoint (Broncus Technologies, Inc., Mountain View, CA, USA), for the diagnosis of small (≤30 mm) peripheral pulmonary lesions (PPL).Between May 2011 and December 2011, we recruited 68 consecutive patients presenting with a PPL 30 mm or less in diameter determined by chest computed tomography. We used the LungPoint system before bronchoscopy to identify the bronchus into which the bronchoscope should be advanced. We used a thin bronchoscope. EBUS-GS was performed using an endoscope ultrasonography system equipped with a 20-MHz mechanical radial-type probe. We used a guide sheath with an external diameter of 1.95 mm, thin forceps and brushing.The diagnostic yield of the 68 PPL was 77.9%; it was 83.7% and 68.0% for the malignant and benign lesions, respectively. Notably, three cases were diagnosed by transbronchial needle-aspiration cytology alone. Univariate and multivariate analyses showed that the EBUS probe localization was the most significant contributor to successful diagnosis (diagnostic yield: within vs adjacent to the lesion = 92.1% vs 60.0%, respectively; P = 0.004 and P = 0.003, respectively).The combination of EBUS-GS and LungPoint was useful for diagnosing small PPL.

切迫流産のため入院を経験した妊婦6名を対象に, 1 心理・行動の経時的変化, 2 経験した心理的喪失と, その回復過程に影響を及ぼした因子を明らかにすることを目的として, 入院初期・退院予測期・妊娠末期・産後1週・産後1か月・2か月に半構成的面接調査を行った.その結果, 妊婦は切迫流産の症状出現あるいは入院によって, 妊娠の継続 (胎児の生存). に関して悲観的予測をし, 一時的に危機状況に陥った. 予期しない出来事を体験しての心理的喪失感として自己不全感を抱いていた. 症状の軽減と胎児の生存と成長の確認を得ることで, この危機状況から回復した. 入院生活では, 薬物療法・安静という制限された生活によって心身のストレスを受け, 妻・母親役割, 職業人としての役割, 友人との交流に関して喪失感を体験していた. しかし, 医療従事者, 家族, 友人, 同室者のサポートが, 心理的喪失感の軽減に影響していた.入院中に胎児の奇形を心配した妊婦の不安は, 出産後, 児が五体満足であることを確認するまで続いていた.妊娠初期の切迫流産の体験は, 時間の経過とともに薄れていき, 特に, 無事分娩を終了し, 育児が順調にいくことで「過去のこと」として整理されていた.

This study demonstrates the feasibility of using a novel 2D spectrum ultrasonic tissue characterization (UTC) technique for prostate-cancer diagnosis. Normalized 2D spectra are computed by performing Fourier transforms along the range (beam) and the cross-range directions of the digital radio-frequency echo data, then dividing by a reference spectrum. This 2D spectrum method provides axial and lateral information of tissue microstructures, an improvement over the current 1D spectrum analysis which only provides axial information. A pilot study was conducted on four prostate-cancer patients who underwent radical prostatectomies. Cancerous and noncancerous regions of interest, identified through histology, were compared using four 2D spectral parameters: peak value and 3 dB width of the radially integrated spectral power (RISP), slope and intercept of the angularly integrated spectral power (AISP). For noncancerous and cancerous prostatic tissues, respectively, our investigation yielded 23 +/- 1 and 26 +/- 1 dB for peak value of RISP, 7.8 +/- 0.5 degrees and 7.6 +/- 0.6 degrees for 3 dB of RISP, -2.1 +/- 0.2 and -2.7 +/- 0.4 dB/MHz for slope of AISP, and 92 +/- 5 and 112 +/- 6 dB for intercept of AISP. Preliminary results indicated that 2D spectral UTC has the potential for identifying tumor-bearing regions within the prostate gland.

The antigenicity of the interferon (IFN) produced in transgenic mice carrying an extra mouse IFN-β gene under the control of mouse metallothionein-I enhancer-promoter was examined after induction with Cd 2+ . Unexpectedly, IFN-α in addition to IFN-β was detected in the serum. Induction of IFN-α was also observed when recombinant mouse IFN-β was injected into normal mice. IFN-a was first detected in the circulation 6–10 hr after the administration of IFN-β, and after 12 hr, IFN-α became the major component of serum IFN. On the other hand, when IFN-a was injected, no production of IFN-β was observed. Messenger RNAs specific for IFN-α and endogenous IFN-β were detected in the spleen, though the amount of IFN-,B mRNA was much less than that of IFN-a mRNA. These mRNAs were not detected in other organs including the liver where exogenous IFN-β gene was markedly expressed. These observations showed that the expression of IFN-α is inducible by IFN-β in the mouse, and the spleen was suggested to be the main site of production. Possible mechanisms of the induction are discussed.

Objective To examine the functional anatomy of the ocular circulation in four bottlenose dolphins (Tursiops truncatus) and five melon-headed whales (Peponocephala electra). Procedure Eyes were obtained postmortem from whales that died while in captivity and/or were found beached. Specimens from whales were investigated using scanning electron microscopy (SEM) of vascular corrosion casts and histology. The thermal characteristics of live dolphin eyes were measured using an infrared imaging system. Results The whale eye receives its primary blood supply from the ophthalmic rete, which extends just behind the eyeball. The ophthalmic rete diverges from the basilar rete and the cervical rete via the posterior thoracic artery. The iris and ciliary processes are supplied by iridic arteries via the major arterial circle that is located around the iris edge. The retinal vessels show the holangiotic type. Choroidal arteries run in parallel arrays so as to interdigitate the densely packed choroidal veins. The venules in the conjunctival fold and palpebral conjunctiva form a well-developed venous plexus. Thermographic examination revealed that the eye shows a substantially higher degree of thermal emission than adjacent skin areas. Conclusions The cetacean eye is characterized by a unique vascular pattern and multivessel plexuses, which are quite different from those of terrestrial mammals. This suggests that the ocular vasculature might function as a thermoregulatory system so that the appropriate operating temperature for the photoreceptors can be maintained in a deep and cold aquatic environment. The distinctive plexuses in the orbit might also be for pooling blood in the eye to conserve oxygen during dives. The ophthalmic rete might play a role in a pressure-damping effect on cetacean ocular circulation as well.

To expand the spectral window for the light-induced charge separation in photosystem (PS) I of oxygenic photosynthetic organisms artificially, Rhodamine B (RhB) was covalently linked to the solutio...

As a rapid and sensitive method to detect canine distemper virus (CDV), an immunocapture enzyme-linked immunosorbent assay (ELISA) was performed. The sensitivity and specificity of the immunocapture ELISA were considered to be high enough. Virus neutralizing (VN) test was also established using the immunocapture ELISA. By using this test, the different cross VN titers between sera of dogs experimentally infected with the Onderstepoort strain and those with a field isolate of CDV were observed.

To analyze the effect of human T-cell leukemia virus type I (HTLV-I) on cellular gene expression and its relation to tumorigenesis, two lines of transgenic mice carrying the long terminal repeat (LTR)-env-pX-LTR regions of the HTLV-I genome were produced. The transgene was expressed in many organs, including the brain, salivary gland, spleen, thymus, skin, muscle, and mammary gland. We found that the expression of the c-fos and c-jun genes, but not of the lyn and c-myc genes, was augmented 2- to 20-fold in histologically normal skin and muscle of these mice. The augmentation was tissue specific, suggesting the involvement of a cellular factor in the transgene action. In these mice, a three to seven times higher incidence of tumors was seen as compared with the control mice. These tumors included mesenchymal tumors, such as fibrosarcoma, neurofibroma, and lipoma, and adenocarcinomas of the mammary gland, salivary gland, and lung. The c-fos and c-jun genes were also activated in these tumors. The possible roles of elevated c-fos and c-jun gene expression in tumorigensis are discussed.

Human T cell leukemia virus type-I (HTLV-I) is the etiologic agent of adult T cell leukemia and has also been suggested to be involved in other diseases such as chronic arthritis or myelopathy. To elucidate pathological roles of the virus in disease, transgenic mice were produced that carry the HTLV-I genome. At 2 to 3 months of age, many of the mice developed chronic arthritis resembling rheumatoid arthritis. Synovial and periarticular inflammation with articular erosion caused by invasion of granulation tissues were marked. These observations suggest a possibility that HTLV-I is one of the etiologic agents of chronic arthritis in humans.

Objective To elucidate the tissue specificity of the expression of HIV-1 genes in an animal and its pathological effects on these tissues. Design and methods Transgenic mice carrying a defective HIV-1 genome were bred in order to overcome the host-range barrier of this virus. Results mRNA specific to the transgene was detected in the eyes and the spleen, and, in smaller quantities, in the thymus and the brain. Interestingly, many of the transgenic mice developed cataracts at 3-6 months of age. Swelling and vacuolation of the lens fiber cells were marked, but the epithelial cells of the lens were less affected. HIV antigens were detected in the lens fiber cells and the retina by immunological staining. Accumulation of large amounts of p24 Gag antigen was demonstrated in the affected lens by immunoblot analysis, while negligible Env or other viral proteins was detected. Although accumulation of the Gag protein was also detected in the skin and the brain, no apparent abnormality was observed in these tissues. Conclusions Preferential expression of the HIV genes in the eyes, skin, brain and lymphoid tissues was demonstrated. The accumulation of the Gag protein is suggested to have detrimental effects on lens fiber cells, causing cataracts.