Characterization of the gut microbiota of Kawasaki disease patients by metagenomic analysis

Kawasaki disease (KD) is an acute febrile illness of early childhood. Previous reports have suggested that genetic disease susceptibility factors, together with a triggering infectious agent, could be involved in KD pathogenesis; however, the precise etiology of this disease remains unknown. Additionally, previous culture-based studies have suggested a possible role of intestinal microbiota in KD pathogenesis. In this study, we performed metagenomic analysis to comprehensively assess the longitudinal variation in the intestinal microbiota of twenty-eight KD patients. Several notable bacterial genera were commonly extracted during the acute phase, whereas a relative increase in the number of Ruminococcus bacteria was observed during the non-acute phase of KD. The metagenomic analysis results based on bacterial species classification suggested that the number of sequencing reads with similarity to five Streptococcus spp. (S. pneumonia, pseudopneumoniae, oralis, gordonii, and sanguinis), in addition to patient-derived Streptococcus isolates, markedly increased during the acute phase in most patients. Streptococci include a variety of pathogenic bacteria and probiotic bacteria that promote human health; therefore, this further species discrimination could comprehensively illuminate the KD-associated microbiota. The findings of this study suggest that KD-related Streptococci might be involved in the pathogenesis of this disease

Homogeneity of the 16S rDNA sequence among geographically disparate isolates of Taylorella equigenitalis

BACKGROUND: At present, six accessible sequences of 16S rDNA from Taylorella equigenitalis (T. equigenitalis) are available, whose sequence differences occur at a few nucleotide positions. Thus it is important to determine these sequences from additional strains in other countries, if possible, in order to clarify any anomalies regarding 16S rDNA sequence heterogeneity. Here, we clone and sequence the approximate full-length 16S rDNA from additional strains of T. equigenitalis isolated in Japan, Australia and France and compare these sequences to the existing published sequences. RESULTS: Clarification of any anomalies regarding 16S rDNA sequence heterogeneity of T. equigenitalis was carried out. When cloning, sequencing and comparison of the approximate full-length 16S rDNA from 17 strains of T. equigenitalis isolated in Japan, Australia and France, nucleotide sequence differences were demonstrated at the six loci in the 1,469 nucleotide sequence. Moreover, 12 polymorphic sites occurred among 23 sequences of the 16S rDNA, including the six reference sequences. CONCLUSION: High sequence similarity (99.5% or more) was observed throughout, except from nucleotide positions 138 to 501 where substitutions and deletions were noted

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

切迫早産予測指標としての妊娠前半期における血清中分泌型免疫グロブリンAおよび首尾一貫感覚

取得学位 : 博士（保健学）, 学位授与番号 : 医博甲第2057号 , 学位授与年月日 : 平成21年9月28日, 学位授与大学 : 金沢大学, 審査結果の報告日 : 平成21年8月26

Effect of Ethanol in Paclitaxel Injections on the Ethanol Concentration in Exhaled Breath

BACKGROUND: Ethanol is included in certain injectable preparations of anticancer drugs to increase their solubility. Since the volume of ethanol in these preparations is approximately half of the total injection volume, the potential inhibitory effects of ethanol on the central nervous system cannot be disregarded, especially considering that patients may drive immediately after administration of the medication. Therefore, the concentration of ethanol was examined in exhaled breath after administration of paclitaxel, an anticancer medication containing ethanol. METHODS: The ethanol concentration in exhaled breath immediately after an intravenous infusion of paclitaxel was measured in 30 patients, using a balloon-type gas detector tube. Correlations between the concentration of ethanol in exhaled breath and the total amount of ethanol administered or the intravenous infusion speed were calculated. RESULTS: The mean ethanol concentration in exhaled breath was 0.028 ± 0.015 mg/L. The correlation between the ethanol concentration in exhaled breath and the total dose of ethanol was weak (R(2) = 0.25; p = 0.055), while the intravenous infusion speed showed a stronger positive correlation with the concentration of ethanol in the breath (R(2) = 0.49; p = 0.11). The maximum concentration of ethanol measured in exhaled breath (0.06 mg/L) was equivalent to 40% of the threshold for drunk driving, as specified in the Road Traffic Act in Japan. CONCLUSION: In this study, no patient had a breath ethanol concentration exceeding the legal threshold for drunk driving. However, it is still advisable for patients to avoid driving after receiving paclitaxel injections. When driving cannot be avoided, patients should wait for a sufficient time after receiving the injection before driving

Mother\u27s feelings of distress and related factors resulting from the crying of her one-month-old infants

金沢大学医薬保健研究域保健学

Corynebacterium ulcerans 0102 carries the gene encoding diphtheria toxin on a prophage different from the C. diphtheriae NCTC 13129 prophage

BACKGROUND: Corynebacterium ulcerans can cause a diphtheria-like illness, especially when the bacterium is lysogenized with a tox gene-carrying bacteriophage that produces diphtheria toxin. Acquisition of toxigenicity upon phage lysogenization is a common feature of C. ulcerans and C. diphtheriae. However, because of a lack of C. ulcerans genome information, a detailed comparison of prophages has not been possible between these two clinically important and closely related bacterial species. RESULTS: We determined the whole genome sequence of the toxigenic C. ulcerans 0102 isolated in Japan. The genomic sequence showed a striking similarity with that of Corynebacterium pseudotuberculosis and, to a lesser extent, with that of C. diphtheriae. The 0102 genome contained three distinct prophages. One of these, ΦCULC0102-I, was a tox-positive prophage containing genes in the same structural order as for tox-positive C. diphtheriae prophages. However, the primary structures of the individual genes involved in the phage machinery showed little homology between the two counterparts. CONCLUSION: Taken together, these results suggest that the tox-positive prophage in this strain of C. ulcerans has a distinct origin from that of C. diphtheriae NCTC 13129

Mother\u27s feelings of distress and related factors resulting from the crying of her one-month-old infants

The purpose of this study was to clarify the nature of mother\u27s distress and its related factors resulting from the crying of her one-month-old infant. Subjects and Methods The subjects were mothers who delivered children in hospitals/maternity clinics in the Hokuriku district and who gave consent to our survey at the time of health examinations for one-month-old infants. The contents of the questionnaire were: characteristics of the infant\u27s crying, states of mother\u27s sleep, feeding and receiving support related to her distress about her infant and its associated factors. Scores were obtained using a 4-point Likert scale. Results Effective responses were obtained from 630 mothers, who consisted of 298 primiparas (47.3%) and 332 multiparas (52.7%). About 50% of the mothers experienced distress because they felt at a loss when their infant cried and when the infant did not stop crying even when being held or lulled. New mothers who had no experience with infants were more likely to indicate distress. The distress was significantly associated with factors such as the way the infant cried and if the infant failed to fall asleep in a timely way. It also was associated with factors in the mother\u27s life such as feeling burdened with childcare and lack of confidence in childcare. Conclusion In order to support mothers having a one-month-old infant, it is important to pay attention to the characteristics of infant\u27s crying, states of mother\u27s fatigue, mother\u27s expression of depression, and to know how mother comprehends childcare and her infant\u27s crying. For screening to be effective it must include both infant and mother factors. 目　的 　生後1 ヶ月児の泣きに対する母親の困難感とその感情に関連する要因を明らかにすることを目的とし た。 対象と方法 　北陸地方の病産院にて出産し，1 ヶ月健診時に調査の同意が得られた母親を対象に，自己記入式質問 紙調査を実施した。調査内容は，児の泣きに対する母親の困難感と，その関連要因として，児の泣きの 性質や母親の睡眠・授乳状況，サポート状況などの質問項目を設定し，各々4段階リカート尺度で点数 化した。 結　果 　有効回答は，初産婦298名（47.3％），経産婦332名（52.7％），合計630名であった。全体の約半数の母親が， 児が泣くと戸惑ったり，抱いたり，あやしても泣きやまない困難な状況を経験していた。困難感を示し た母親は，小さな子どもと接したことのない初産婦に多く，子どもの泣き方が特徴的であったり，なか なか寝入らないなど，子ども側の要因と母親の生活状況，育児に対する負担感や自信感等の母親側の要 因が困難感に関連していた。 結　論 　生後1 ヶ月時の母児の支援には，児側の要因と母親側の要因の双方に着目し，児の泣きの特徴や，母 親の疲労状態，育児に対する気持ち等に注意を向け，母親が児の泣きをどのようにとらえているのかを 知ることが重要であり，これらのスクリーニングの必要性が示唆された

Mother\u27s feelings of distress and related factors resulting from the crying of her one-month-old infants

金沢大学医薬保健研究域　保健学

Characterization of Quasispecies of Pandemic 2009 Influenza A Virus (A/H1N1/2009) by De Novo Sequencing Using a Next-Generation DNA Sequencer

Pandemic 2009 influenza A virus (A/H1N1/2009) has emerged globally. In this study, we performed a comprehensive detection of potential pathogens by de novo sequencing using a next-generation DNA sequencer on total RNAs extracted from an autopsy lung of a patient who died of viral pneumonia with A/H1N1/2009. Among a total of 9.4×106 40-mer short reads, more than 98% appeared to be human, while 0.85% were identified as A/H1N1/2009 (A/Nagano/RC1-L/2009(H1N1)). Suspected bacterial reads such as Streptococcus pneumoniae and other oral bacteria flora were very low at 0.005%, and a significant bacterial infection was not histologically observed. De novo assembly and read mapping analysis of A/Nagano/RC1-L/2009(H1N1) showed more than ×200 coverage on average, and revealed nucleotide heterogeneity on hemagglutinin as quasispecies, specifically at two amino acids (Gly172Glu and Gly239Asn of HA) located on the Sa and Ca2 antigenic sites, respectively. Gly239 and Asn239 on antigenic site Ca2 appeared to be minor amino acids compared with the highly distributed Asp239 in H1N1 HAs. This study demonstrated that de novo sequencing can comprehensively detect pathogens, and such in-depth investigation facilitates the identification of influenza A viral heterogeneity. To better characterize the A/H1N1/2009 virus, unbiased comprehensive techniques will be indispensable for the primary investigations of emerging infectious diseases