High-Throughput Transposon Mutagenesis of Corynebacterium glutamicum and Construction of a Single-Gene Disruptant Mutant Library

A simple and high-throughput transposon-mediated mutagenesis system employing two different types of transposons in combination with direct genomic DNA amplification and thermal asymmetric interlaced PCR (TAIL-PCR) was developed. Each of the two minitransposons based on IS31831 (ISL3 family) and Tn5 (IS4 family) was integrated into the Corynebacterium glutamicum R genome. By using BLAST and Perl, transposon insertion locations were automatically identified based on the sequences of TAIL-PCR products of mutant cells. Insertion locations of 18,000 mutants were analyzed, and a comprehensive insertion library covering nearly 80% of the 2,990 open reading frames of C. glutamicum R was generated. Eight thousand of the mutants, exhibiting disruption in 2,330 genes, survived on complex medium under normal laboratory conditions, indicating that the genes were not essential for cell survival. Of the 2,330 genes, 30 exhibited high similarity to essential genes of Escherichia coli or Bacillus subtilis. This approach could be useful in furthering genetic understanding of cellular life and facilitating the functional analysis of microorganisms

A survey of current status of patient education and willingness to use disease management service programs in hospitals and clinics in Japan

糖尿病、虚血性心疾患及び心不全について、わが国の医療機関での患者教育の実態及び問題点を把握し、患者教育の方向性を検討、さらに、今後、外部疾病管理サービスの受入れについて医療機関の意識や障害を明らかにすることを目的に調査を行った。内科を標榜する一般病床をもつ病院を病床規模ごとに全国から1,000病院、診療所100ヵ所を対象に、無記名自記式選択式質問紙による郵送回収調査を行った。322病院、26診療所を分析対象とした。結果、専門医、専門スタッフ、専門外来、患者教育プログラムの整備状況は病院規模が大きくなるほど高く、患者教育については200床未満の病院・診療所の実施割合は低く、内容も限定的であった。患者教育の問題点についてはスタッフ数・専門スタッフ不足及び看護師による教育が診療報酬対象にならないことが上げられ、5～6割の病院がこれを補完するための外部疾病管理サービスは条件が整えば採用したい意向を示した。We conducted a survey of patient education on diabetes mellitus, ischemic heart disease and heart failure, in order to identify the current status and problems concerning patient education at hospitals and clinics in Japan, to determine the future directions for patient education, and to explore the willingness and impediment of hospitals and clinics to accept external disease management service programs in the future. We registered 1000 hospitals and 100 clinics according to the number of beds from all over Japan among those with a department of internal medicine with a proportion of beds allocated to general patients. We sent anonymous self-administered selective questionnaires to these institutions and collected the responses by mail. Finally, 322 hospitals and 26 clinics were included for the analysis. Larger hospitals tended to be better prepared in

Discovery of Tetrasubstituted Imidazolines as Potent and Selective Neuropeptide Y Y5 Receptor Antagonists: Reduced Human Ether-a-go-go Related Gene Potassium Channel Binding Affinity and Potent Antiobesity Effect

A series of novel imidazoline derivatives was synthesized and evaluated as neuropeptide Y (NPY) Y5 receptor antagonists. Optimization of previously reported imidazoline leads, <b>1a</b> and <b>1b</b>, was attempted by introduction of substituents at the 5-position on the imidazoline ring and modification of the bis(4-fluorphenyl) moiety. A number of potent derivatives without human ether-a-go-go related gene potassium channel (hERG) activity were identified. Selected compounds, including <b>2a</b>, were shown to have excellent brain and CSF permeability. Compound <b>2a</b> displayed a suitable pharmacokinetic profile for chronic in vivo studies and potently inhibited d-Trp³⁴NPY-induced acute food intake in rats. Oral administration of <b>2a</b> resulted in a potent reduction of body weight in a diet-induced obese mouse model