新興・再興感染症対応のための基礎的応用的研究の展開（仮称）

Preliminary basic and applied research on a project on emerging and re-emerging infectious diseases has been carried out as part of an application project to the High Tech Research Center from November 2007 to March 2008. During this period, Anisakis, Paragonimus, Cryptosporidium, Severe invasive Streptococcus, Campylobacter, Helicobacter, Salmonella, Enterohemorrhagic Escherichia coli O157:H7, Enteric virus, Borna disease virus and some other pathogenic organisms, were examined in conjunction with several investigative modalities; including molecular biology, cell biology, microbiology, immunology, virology (where applicable), pathobiology, as well as epidemiology. Our aim is that this core project will act basis of a strong and vibrant center in the field of emerging and re-emerging infectious diseases in our university, as well as providing our undergraduate and postgraduate students with the research opportunities and experience in this area, which will allow them to become leading experts in the control of infectious diseases for the benefit of society

Identification and characterization of intervening sequences within 23S rRNA genes from more than 200 Campylobacter isolates from seven species including atypical campylobacters

<p>Abstract</p> <p>Background</p> <p>Identification and characterization of intervening sequences (IVSs) within 23S rRNA genes from <it>Campylobacter </it>organisms including atypical campylobacters were carried out using two PCR primer pairs, designed to generate helix 25 and 45 regions.</p> <p>Results</p> <p>Only <it>C. sputorum </it>biovar sputorum LMG7975 and fecalis LMG8531, LMG8534 and LMG6728 of a total of 204 <it>Campylobacter </it>isolates (n = 56 <it>C. jejuni</it>; n = 11 <it>C. coli</it>; n = 33 <it>C. fetus</it>; n = 43 <it>C. upsaliensis</it>; n = 30 <it>C. hyointestinalis</it>; n = 4 <it>C. sputorum </it>biovar sputorum; n = 5 <it>C. sputorum </it>biovar fecalis; n = 5 <it>C. sputorum </it>biovar paraureolyticus; n = 10 <it>C. concisus</it>; n = 7 <it>C. curvus</it>) were shown to carry IVSs in helix 25 region. <it>C. sputorum </it>biovar fecalis LMG8531 and LMG8534, interestingly, carried two different kinds of the 23S rRNA genes with and without the IVS, respectively. Consequently, in a total of 265 isolates of 269, including 65 <it>C. lari </it>isolates examined previously, the absence of IVSs was identified in the helix 25 region. In the helix 45 region, all the <it>C. hyointestinalis</it>, <it>C. sputorum </it>and <it>C. concisus </it>isolates were shown not to carry any IVSs. However, the 30 of 56 <it>C. jejuni </it>isolates (54%), 5 of 11 <it>C. coli </it>(45%), 25 of 33 <it>C. fetus </it>(76%), 30 of 43 <it>C. upsaliensis </it>(70%) and 6 of 7 <it>C. curvus </it>(90%) were shown to carry IVSs. In <it>C. jejuni </it>and <it>C. upsaliensis </it>isolates, two different kinds of the 23S rRNA genes were also identified to occur with and without IVSs in the helix 45 region, respectively.</p> <p>Conclusions</p> <p>Secondary structure models were also constructed with all the IVSs identified in the present study. In the purified RNA fractions from the isolates which carried the 16S or 23S rRNA genes with the IVSs, no 16S or 23S rRNA was evident, respectively.</p

Molecular characterization of the sequences of the 16S-23S rDNA internal spacer region (ISR) from isolates of Taylorella asinigenitalis

<p>Abstract</p> <p>Background</p> <p>Sequence information on the 16S-23S rDNA internal spacer region (ISR) exhibits a large degree of sequence and length variation at both the genus and species levels. A primer pair for the amplification of 16S-23S rDNA ISR generated three amplicons for each of isolates of <it>Taylorella asinigenitalis </it>(UCD-1^T, UK-1 and UK-2).</p> <p>Findings</p> <p>Following TA cloning and sequencing, the three isolates of <it>T. asinigenitalis </it>were demonstrated to possess three ISR units of different lengths. Although the three corresponding ISRs (A, B and C) were identified to be identical to each other (UK-1 and UK-2 isolates), the ISRs shared approximately 95.3–98.9% nucleotide sequence similarities between the UCD-1^Tand UK-1/-2 isolates. A typical order of two intercistronic tRNA genes (5'-tRNA^Ile-tRNA^Ala-3') with the different nucleotide spacers [44 through 51 base pairs (bp)] in length was identified among the isolates. The consensus sequences of the antiterminators of <b>boxB </b>and <b>boxA </b>were also identified in all ISRs. Thus, three ISRs were identified for each isolate, and therefore, at least three distinctly different ribosomal RNA operons were suggested to occur in the genome of <it>T. asinigenitalis</it>. This was also confirmed by Southern hybridization procedure.</p> <p>Conclusion</p> <p>The present study represents a dendrogram constructed based on the nucleotide sequence data of 16S-23S rDNA ISR for <it>T. asinigenitalis</it>, which may aid in the phylogenetic positioning of <it>T. asinigenitalis </it>within the genus <it>Taylorella</it>, and in the molecular discrimination of <it>T. asinigenitalis</it>.</p

Sol-gel chemistry mediated Zn/Al-based complex dispersant for SWCNT in water without foam formation

We report a bimetallic Zn/Al complex as an efficient inorganic dispersant for SWCNT, synthesized from Zn(CH3COO)(2) and Al(NO3)(3). The Zn/Al complex shows more than four times greater efficiency at dispersing SWCNT than widely used surfactants (CTAB and SDS). Besides remarkable dispersibility, the Zn/Al complex does not foam upon any shaking treatment and it can be used just after quick dissolution of the powdered form, which is a marked advantage over surfactants. The Zn/Al complex, containing amorphous Al(CH3COO)(3) and a complex of Zn2+ and NO3- ions, should have a unique dispersion mechanism, differing from the surfactants. Al(CH3COO)(3) has higher affinity for SWCNT than ions, adsorbing onto its surface in the first layer and attracting Zn2+ and NO3- ions. Charge transfer interactions between the Zn/Al complex and SWCNT, as evidenced by optical absorption spectroscopy, should induce a charge on SWCNT; the zeta potential of such coated SWCNT was +55 mV, indicating a high dispersion stability in aqueous media. Hence, the Zn/Al complex can widen the applications of SWCNT to various technologies such as the transparent and conductive films, as well as high performance composite polymers. (C) 2015 Elsevier Ltd. All rights reserved.ArticleCARBON. 94:518-523 (2015)journal articl

Ultrasonographic Changes of the Knee Joint Reflect Symptoms of Early Knee Osteoarthritis in General Population; The Nagahama Study

Objective: Radiographic changes in knee osteoarthritis (OA) are not always associated with symptoms, especially in its early stages. Ultrasonography (US) can detect early changes in the knee joint, but the changes that reflect symptoms have not been fully elucidated. This study aimed to identify US-detectable changes in the knee that are often associated with knee symptoms and demonstrate the feasibility of early diagnosis in symptomatic knee OA using US. Design: In this cross-sectional community-based study, 1, 667 participants aged ≥60 years (1, 103 women [66%]) were included. All participants concurrently underwent US and radiography of the knee and completed the Knee Society Knee Scoring System (KSS) questionnaire. Simple and multiple regression analyses were used to examine the associations between US findings and KSS symptom subscales. Results: Among all participants, medial meniscus protrusion and medial osteophytes, age, and body mass index showed significant associations with KSS symptom scores. Among 894 participants with Kellgren-Lawrence (KL) grade ≤1, medial osteophytes and age were significantly associated with KSS symptom score. US measures were more related to KSS symptoms than KL grades. Conclusions: Among the knee US-detectable changes, medial osteophytes were strongly associated with knee symptoms. Osteophytes are reliable predictors of symptomatic early knee OA, even in participants with few radiographic OA changes

ウレアーゼ陽性高温性カンピロバクター（UPTC）のウレアーゼ遺伝子オペロンの分子解析

When cloning, sequencing and characterization of the genetic organization of urease genes within urease-positive thermophilic Campylobacter (UPTC) were carried out, an approximate 5.1 kilo base pair region encoding a urease gene operon was identified with recombinant plasmid DNAs from a genomic DNA library of a Japanese isolate (CF89-12). Six closely spaced and putative open reading frames (ORFs) for ureA, ureB, ureE, ureF, ureG and ureH were detected. ATG codons initiated each ORF of the UPTC urease operon except for ureB and ureH, which commenced with the most probable TTG codon. Overlaps were detected between ureA and ureB and also between ureB and ureE. Probable ribosome-binding sites and a putative p independent transcriptional termination region were identified. Two putative promoter structures, consisting of consensus sequences at the -35 and -10 like regions were also identified. Construction of a neighbor-joining tree based on the nucleotide sequence data of urease genes indicated that UPTC formed a cluster with some Helicobacter organisms separate from the other urease-producing bacteria, suggesting a commonly shared ancestry between UPTC and Helicobacter urease genes

Poly[[hexa-μ-cyanido-manganese(II)iron(III)] penta­hydrate]

The structure of the title compound, MnII[FeIII(CN)6]2/3·5H2O, features a face-centered cubic –Mn—NC—Fe– framework with both Mn and Fe having site symmetry m m. Since one-third of the [Fe(CN)6]3− units are missing for a given formula in order to maintain charge neutrality, each Mn atom around such a vacancy is coordinated not only by the N atoms of the CN groups but also by the O atoms of the ligand water mol­ecules. In addition to ligand water mol­ecules, two types of non-coordinated water mol­ecules, so-called zeolitic water mol­ecules, exist in the inter­stitial sites of the –Mn—NC—Fe– framework. The positions of the O atoms of the zeolitic water mol­ecules are fixed by the linkage via hydrogen bonds between ligand water and zeolitic water mol­ecules. The structure is related to a recently reported rubidium manganese hexa­cyano­ferrate. Site occupancy factors for Fe, C, N are 0.67; for two O atoms the value is 0.83 and for one O atom is 0.17

A two-step species-specific 16S rRNA PCR assay for the detection of Taylorella equigenitalis in horses

<p/> <p>A two-step PCR assay was developed for the molecular detection of Taylorella equigenitalis, a Gram-negative genital bacterial pathogen in horses. Two specific oligonucleotide primers (TE16SrRNABCHf [25mer] and TE16SrRNABCHr [29mer]) were designed from multiple alignments of the 16S rRNA gene loci of several closely related taxa, including T. asinigenitalis. Subsequent enhanced surveillance of 250 Thoroughbred animals failed to detect the presence of this organism directly from clinical swabs taken from the genital tract of mares and stallions. Such a molecular approach offers a sensitive and specific alternative to conventional culture techniques, and has the potential to lead to improved diagnosis and subsequent management of horses involved in breeding programmes.</p

A VCP modulator, KUS121, as a promising therapeutic agent for post-traumatic osteoarthritis

京大開発の薬剤「KUS121」の変形性膝関節症への効果を確認 --外傷性変形性関節症の治療薬として臨床応用へ--. 京都大学プレスリリース. 2020-12-17.Post-traumatic osteoarthritis (PTOA) is a major cause which hinders patients from the recovery after intra-articular injuries or surgeries. Currently, no effective treatment is available. In this study, we showed that inhibition of the acute stage chondrocyte death is a promising strategy to mitigate the development of PTOA. Namely, we examined efficacies of Kyoto University Substance (KUS) 121, a valosin-containing protein modulator, for PTOA as well as its therapeutic mechanisms. In vivo, in a rat PTOA model by cyclic compressive loading, intra-articular treatments of KUS121 significantly improved the modified Mankin scores and reduced damaged-cartilage volumes, as compared to vehicle treatment. Moreover, KUS121 markedly reduced the numbers of TUNEL-, CHOP-, MMP-13-, and ADAMTS-5-positive chondrocytes in the damaged knees. In vitro, KUS121 rescued human articular chondrocytes from tunicamycin-induced cell death, in both monolayer culture and cartilage explants. It also significantly downregulated the protein or gene expression of ER stress markers, proinflammatory cytokines, and extracellular-matrix-degrading enzymes induced by tunicamycin or IL-1β. Collectively, these results demonstrated that KUS121 protected chondrocytes from cell death through the inhibition of excessive ER stress. Therefore, KUS121 would be a new, promising therapeutic agent with a protective effect on the progression of PTOA

Sol–gel chemistry mediated Zn/Al-based complex dispersant for SWCNT in water without foam formation

We report a bimetallic Zn/Al complex as an efficient inorganic dispersant for SWCNT, synthesized from Zn(CH3COO)(2) and Al(NO3)(3). The Zn/Al complex shows more than four times greater efficiency at dispersing SWCNT than widely used surfactants (CTAB and SDS). Besides remarkable dispersibility, the Zn/Al complex does not foam upon any shaking treatment and it can be used just after quick dissolution of the powdered form, which is a marked advantage over surfactants. The Zn/Al complex, containing amorphous Al(CH3COO)(3) and a complex of Zn2+ and NO3- ions, should have a unique dispersion mechanism, differing from the surfactants. Al(CH3COO)(3) has higher affinity for SWCNT than ions, adsorbing onto its surface in the first layer and attracting Zn2+ and NO3- ions. Charge transfer interactions between the Zn/Al complex and SWCNT, as evidenced by optical absorption spectroscopy, should induce a charge on SWCNT; the zeta potential of such coated SWCNT was +55 mV, indicating a high dispersion stability in aqueous media. Hence, the Zn/Al complex can widen the applications of SWCNT to various technologies such as the transparent and conductive films, as well as high performance composite polymers. (C) 2015 Elsevier Ltd. All rights reserved.ArticleCARBON. 94:518-523 (2015)journal articl