Cloning, Sequencing, and Functional Analysis of the Biosynthetic Gene Cluster of Macrolactam Antibiotic Vicenistatin in Streptomyces halstedii

AbstractVicenistatin, an antitumor antibiotic isolated from Streptomyces halstedii, is a unique 20-membered macrocyclic lactam with a novel aminosugar vicenisamine. The vicenistatin biosynthetic gene cluster (vin) spanning ∼64 kbp was cloned and sequenced. The cluster contains putative genes for the aglycon biosynthesis including four modular polyketide synthases (PKSs), glutamate mutase, acyl CoA-ligase, and AMP-ligase. Also found in the cluster are genes of NDP-hexose 4,6-dehydratase and aminotransferase for vicenisamine biosynthesis. For the functional confirmation of the cluster, a putative glycosyltransferase gene product, VinC, was heterologously expressed, and the vicenisamine transfer reaction to the aglycon was chemically proved. A unique feature of the vicenistatin PKS is that the loading module contains only an acyl carrier protein domain, in contrast to other known PKS-loading modules containing certain activation domains. Activation of the starter acyl group by separate polypeptides is postulated as well

Three-dimensional analysis of perineural invasion in extrahepatic cholangiocarcinoma using tissue clearing

Perineural invasion (PNI) is a characteristic invasion pattern of distal cholangiocarcinoma (DCC). Conventional histopathologic examination is a challenging approach to analyze the spatial relationship between cancer and neural tissue in full-thickness bile duct specimens. Therefore, we used a tissue clearing method to examine PNI in DCC with three-dimensional (3D) structural analysis. The immunolabeling-enabled 3D imaging of solvent-cleared organs method was performed to examine 20 DCC specimens from five patients and 8 non-neoplastic bile duct specimens from two controls. The bile duct epithelium and neural tissue were labeled with CK19 and S100 antibodies, respectively. Two-dimensional hematoxylin/eosin staining revealed only PNI around thick nerve fibers in the deep layer of the bile duct, whereas PNI was not identified in the superficial layer. 3D analysis revealed that the parts of DCC closer to the mucosa exhibited more nerves than the normal bile duct. The nerve fibers were continuously branched and connected with thick nerve fibers in the deep layer of the bile duct. DCC formed a tubular structure invading from the epithelium and extending around thin nerve fibers in the superficial layer. DCC exhibited continuous infiltration around the thick nerve fibers in the deep layer. This is the first study using a tissue clearing method to examine the PNI of DCC, providing new insights into the underlying mechanisms

高齢者における歯科治療前の不整脈

Seventy-five dental patients between the ages of 65 and 94, who had previously been diagnosed as having cardiovascular diseases (CVD), were observed between January 1995 and August 1998. The visits were conducted either in the hospital or at home. There were a total of 477 patient-visits, and at each patient-visit, an ECG (lead II) was monitored for 5 minutes as part of a general health assessment before dental treatment. As a result, various arrhythmias were found. One patient was prescribed an antiarrhythmic drug because of ventricular bigeminy. Dental treatment was deferred for two patients because of multiple VPC or sinus tachycardia. In this study, we found that a potentially serious arrhythmia in patients with no history of arrhythmia, and routine pre-operative assessments including an ECG are recommended

介助磨きに対する子どもの行動

Clarification of the normal developmental stages with regard to recumbent-position brushing is considered important for the purpose of giving appropriate guidance in teeth cleaning infants. In order to identify the ages at which children may accept recumbent-position brushing, we had a number of mothers perform recumbent-position brushing upon normal preschool children (ages 0-6 years), and observed the children\u27s behavior. 79 children (79.0%), classified as the adaptive group, were seen to have no negative reaction or behavior whatsoever to recumber-position brushing. However, 21 children (21.0%) composed an unadaptive group in which some rejective behavior was observed. As a result of analyses by AIC, the calender age which thus distinguishes the adaptive group from the unadaptive group was 2 years and 6 months. It was concluded that children 2 years and 6 months or older are adaptive to recumbent-position brushing while those less than 2 years and 6 months are unadaptive

CIPRO 2.5: Ciona intestinalis protein database, a unique integrated repository of large-scale omics data, bioinformatic analyses and curated annotation, with user rating and reviewing functionality

The Ciona intestinalis protein database (CIPRO) is an integrated protein database for the tunicate species C. intestinalis. The database is unique in two respects: first, because of its phylogenetic position, Ciona is suitable model for understanding vertebrate evolution; and second, the database includes original large-scale transcriptomic and proteomic data. Ciona intestinalis has also been a favorite of developmental biologists. Therefore, large amounts of data exist on its development and morphology, along with a recent genome sequence and gene expression data. The CIPRO database is aimed at collecting those published data as well as providing unique information from unpublished experimental data, such as 3D expression profiling, 2D-PAGE and mass spectrometry-based large-scale analyses at various developmental stages, curated annotation data and various bioinformatic data, to facilitate research in diverse areas, including developmental, comparative and evolutionary biology. For medical and evolutionary research, homologs in humans and major model organisms are intentionally included. The current database is based on a recently developed KH model containing 36 034 unique sequences, but for higher usability it covers 89 683 all known and predicted proteins from all gene models for this species. Of these sequences, more than 10 000 proteins have been manually annotated. Furthermore, to establish a community-supported protein database, these annotations are open to evaluation by users through the CIPRO website. CIPRO 2.5 is freely accessible at http://cipro.ibio.jp/2.5

CIPRO 2.5: Ciona intestinalis Protein integrated database with large-scale omics data, bioinformatic analyses and curated annotation, with ability for user rating and comments

CIPRO database is an integrated protein database for a tunicate species Ciona intestinalis that belongs to the Urochordata. Although the CIPRO database deals with proteomic and transcriptomic data of a single species, the animal is considered unique in the evolutionary tree, representing a possible origin of the vertebrates and is a good model for understanding chordate evolution, including that of humans. Furthermore, C. intestinalis has been one of the favorites of developmental biologists; there exists a huge amount of accumulated knowledge on its development and morphology, in addition to the recent genome sequence and gene expression data. The CIPRO database is aimed at not only collecting published data, but also presenting unique information, including the unpublished transcriptomic and proteomic data and human curated annotation, for the use by researchers in broad research fields of biology and bioinformatics

The Human Anatomic Gene Expression Library (H-ANGEL), the H-Inv integrative display of human gene expression across disparate technologies and platforms

The Human Anatomic Gene Expression Library (H-ANGEL) is a resource for information concerning the anatomical distribution and expression of human gene transcripts. The tool contains protein expression data from multiple platforms that has been associated with both manually annotated full-length cDNAs from H-InvDB and RefSeq sequences. Of the H-Inv predicted genes, 18 897 have associated expression data generated by at least one platform. H-ANGEL utilizes categorized mRNA expression data from both publicly available and proprietary sources. It incorporates data generated by three types of methods from seven different platforms. The data are provided to the user in the form of a web-based viewer with numerous query options. H-ANGEL is updated with each new release of cDNA and genome sequence build. In future editions, we will incorporate the capability for expression data updates from existing and new platforms. H-ANGEL is accessible at http://www.jbirc.aist.go.jp/hinv/h-angel/

Factor of increased propofol dosage during dental treatment under intravenous anesthesia.

Introduction　Dental treatment using propofol (trade name Diprivan) was carried out for people with disabilities who have difficulties with dental treatment so that dental treatment could be carried out safely and smoothly. This time, we investigated for the purpose of clarifying the factors of those who need 3.5 µg/ml or more when sedated with propofol.　Object and method　Methods　　The subjects were 34 persons, who required the intravenous anesthesia method at the time of dental treatment, among patients who visited Matsumoto Dental University Hos-pital special specialist department from May 1st to September 30 th , 2015. Gender, age, presence or absence of regular medications were noted, weight and height were measured, and BMI was calculated. Disease, treatment, type of disability, and intellectual level from the　medical record were entered on the survey form. As an evaluation of adaptability to dental treatment, they were classified into 4 stages: “Can sit on the medical table”; “Can sleep the 　treatment table”; “Can do an oral examination”; and “Can do PTC”.　The diffuser TCI function was used to initiate intravenous administration at the target 　blood concentration of 3.0 µg/ml. Dental treatment was started with blood concentration in the brain when the opening device was smoothly inserted. If this target was not possible,　0.2 µg/ml each was listed. Blood concentrations and intracerebral concentrations of propofol during treatment were recorded. The intracerebral concentration where the opening device was smoothly inserted, the lowest brain concentration and the maximum brain concentration at the time of treatment were also recorded.Result　34 subjects (28 males, 6 females) had an average age of 37.6±12.4 years. The items asso-　ciated with brain concentration of propofol were subjects aged 50 years or older (P = 0.01), 　BMI (P = 0.15), dental phobia (P = 0.001), and autistic spectrum disease (P = 0.07). The factor that required propofol brain concentration of over 3.5 µg/ml was dental phobia (odds ratio: 28.5: confidence interval 1.₉–421) by the logistic regression analysis.　Conclusion　A factor that requires propofol of 3.5 µg/ml or more at the time of dental treatment, for those who can assume a supine position without making refusing actions at the medical table, was dental phobia. There was no relevance to the content of treatment or to the adaptability to dental treatment.