Analysis of the protein profiles of the antibiotic-resistant Salmonella typhimurium definitive phage type (dt) 104

The emergent Salmonella typhimurium definitive phage type (DT) 104 is of particular global concern due to its frequent isolation and multiple antibiotic resistances. There is thus a need to know the kind of proteins expressed by S. typhimurium DT104 so as to provide a basis for developing an intervention. This study examined the protein profiles of a few isolates of S. typhimurium DT104 and a non-DT104 strain S. typhimurium L1388 (ST). Crude SDS-soluble whole cell and outer membrane protein (OMP) extracts revealed similar protein profiles for both phage types. A single major protein band (28.4 kDa) was observed in periplasmic fractions from both phage types. However, proteins released into growth medium was variable; one of the DT104 isolates had common proteins with the non-DT104 strain ST. Similar SDS-soluble whole cell protein profiles were observed for both phage types grown in a low-iron Medium A at 37oC; but a 38.5 kDa protein (observed in TSB-grown cells) was observed only in the temperature-tolerant DT104 isolate. The protein contents of cell-free ultracentrifuge supernatants of sonically disrupted cells of each of the DT104 isolates were significantly (P < 0.05) more than that from ST L1388, but the latter expressed a 51-kDa protein absent in the supernatants of all DT104 isolates. The higher protein content of DT104s provides possible indication of increased production of protein-like metabolites. Although the N-terminal sequence of the first twenty amino acids of the 51-kDa protein (Ala-Gln-Val-Ile-Asn-Thr-Asn-Ser-Leu-Ser-Leu-Leu-Thr-Gln-Trp-Ala-Ala-Ala-Ala-Ala) showed 14-amino acid overlap and resemblance with the flagillin, FLIC, only fourteen of its 104 trypsin digests were homologous with those of FLIC. Further work is being done to characterize this protein and to investigate its potential for use as vaccine target through antigenicity tests.African Journal of Biotechnology Vol. 4 (7), pp. 727-737, 200

高齢者気管支喘息における気道過敏性と温泉療法

Clinical effects of spa therapy were examined in 150 patients with asthma in relation tobronchial hyperresponsiveness and patient age. 1. The efficacy rate of spa therapy was larger as the patient age was higher: the rate was 73.3% in patients under age 49, 81.8% in those between the ages of 50 and 59, 86.4% in those between the ages of 60 and 69, and 90.6% in those over age 70. The mean of efficacy rates was 83.3% in all subjects. 2. The bronchial hyperresponsiveness (BH) was lower as patient age was higher: the BH in patients between the ages of 60 and 69 and in those over age 70 was significantly lower compared to the BH in those under age 49 (p < O.OOl). 3. Clinical effects of spa therapy tended to be lower in patients with increased bronchial hyperresponsiveness. The bronchial hyperresponsiveness showed a tendency to decrease after spa therapy in whom the therapy was effective, however, the BH did not change in patients with slight or no efficacy during spa therapy.1.温泉療法では,年齢が高くなるほどその有効率も高くなると言う傾向が見られ､49才以下の症例では73.3% ,50-59才の症例では81.8% , 60-69才の症例では86.4% ,70才以上では90.6% であり,全症例の平均有効率は73.3% であった｡　 2.気道過敏性は,年齢が高くなるほど低下する傾向が見られ,60-69才および70才以上の症例の気道過敏性は,49才以下の症例と比べ有 意に低い値を示した(P < 0.001)。　 3.温泉療法の臨床効果は,気道過敏性が強くなるにつれて低下する傾向が見られた｡また,温泉療法の著効例や有効例では,治療により 気道過敏性が低下してくるが､やや有効例や無効例では,気道過敏性はほとんど変化しないことが示された

Termination of Graphene Edges Created by Hydrogen and Deuterium Plasmas

Edge engineering is important for both fundamental research and applications as the device size decreases to nanometer scale. This is especially the case for graphene because a graphene edge shows totally different electronic properties depending on the atomic structure and the termination. It has recently been shown that an atomically precise zigzag edge can be obtained by etching graphene and graphite using hydrogen (H) plasma. However, edge termination had not been studied directly. In this study, termination of edges created by H-plasma is studied by high-resolution electron energy loss spectroscopy (HREELS) to show that the edge is $\mathrm{sp}^{2}$ bonded and the edge carbon atom is terminated by only one H atom. This suggests that an ideal zigzag edge, which is not only atomically precise but also $\mathrm{sp}^{2}$ bonding, can be obtained by H-plasma etching. Etching of the graphite surface with plasma of a different isotope, deuterium (D), is also studied by scanning tunneling microscopy (STM) to show that D-plasma anisotropically etches graphite less efficiently, although it can make defects more efficiently, than H-plasma.Comment: 8 pages, 5 figure

サーモグラフィーによる体表面温度の測定 3.体表面温度の回復率と末消血流量との相関

The body surface peripheral circulation in 12 cases,　including 9 patients with　diabetes mellitus who were suffering coldness, numbness or pain in their feet, and 3 healthy volunteers was examined using Laser-Doppler blood flowmetry. At the same time, the body surface temperature was estimated by thermography. Thermographic results were analyzed quantitatively by calculating a recovery ratio as : Recovery ratio = [Total counts of thermography(Pixels) over temperature (T) after cold loading] ÷ [Initial counts over T before cold loading] x 100(%). The recovery ratio and the blood flow were correlated, r=0.68, p<O.01. The peripheral circulation of 16 patients with diabetes mellitus was observed at three different conditions including, l)placed at room temperature at 20℃for 15 min, 2) submerged and warmed for 5 min in a hot bath at 36℃(i.e. hot loading), and 3) submerged and cooled for 5 min in a water bath at 20℃ (i.e. cold loading). Three different baseline temperatures, 26℃, 27℃ and 28 ℃, were used in processing the　thermographic results into pictures. The highest correlation (r=0.59, p=0.0002) was obtained under the condition of cold loading using a baseline temperature limitation of27℃.The difference ratio (%) of blood flow was calculated as the blood flow at cold loading divided by the blood flow at hot loading in these 16 patients. The difference　ratio of the blood flow and the recovery ratio of thermography were correlated, r=0.46, p<O.OOO1. We found a strong correlation between the results of Laser-Doppler blood flowmetry and one of thermographic methods used to monitor peripheral circulation in patients with diabetes mellitus. Cold loading using a baseline temperature limitation of 27℃ were recommended for further examinations. Patients with low blood flow as well as　with large differences in their peripheral circulation between cold loading and hot　loading had severe coldness in their body surface temperature. We showed the usefulness of the results of thermography, when quantified by picture processing using computer software, in relation with the results of Laser-Doppler blood flowmetry.末梢神経障害を有する糖尿病患者の末梢循環障害の程度を数量的に検討する目的で､下肢に冷感ならびにしびれ感または疼痛を訴える糖尿病患者9症例と健常ボランティア3例の計12例(平均年齢59歳)についてサーモグラフィーを用いて体表面温度を測定した｡更に,サーモグラフィーで得られた結果と末梢皮膚血流量をレーザードプラー血流計を用いて測定して得られた結果と比較した｡サーモグラフィーによる測定で得られた結果は回復率として数量化して表示された｡回復率の算出方法は回復率= [冷水負荷後の特定温度27℃以 上の体表面温度のサーモグラフィーのPixelの総数]÷ [冷水負荷前の特定温度27℃以上の体表面温度のサーモグラフィーのPixelの総数]×100%で求めた｡レーザードプラ-血流計を用いて測定して得られた末梢血流量は左右それぞれ5カ所,計10カ所の測定値の平均で表示した｡その結果,末梢皮膚温度の回復率と末梢皮膚の血流量との問には正の相関関係(r=0.68,p<0.01)が認められた｡次に,末梢皮膚血流量について,室温20℃安静15分後,温水36℃浸水負荷10分後,冷水20℃浸水負荷30分後の異なる3条件について､またサーモグラフィーで得られた結果を,画像処理の過程で用いられた,26℃,27℃,28℃の3つの異なる特定温度との関連について検討を行なった｡対象は,下肢に冷感ならびにしびれ感または疼痛を訴える糖尿病患者16症例(平均年齢69歳,平均HbAIC9.6%)について測定した｡その結果,末梢皮膚 血流量は冷水20℃浸水負荷30分後に測定して得られた結果と,回復率は特定温度27℃で画像処理して得られた結果とが最も相関が高い(r=0.59,p=0.0002)ことが示された｡`次に,相関が高い条件は,室温20℃安静15分後に血流量を測定した場合(r=0.483,p=0.0002)であった｡そして温 水36℃浸水負荷10分後に測定して得られた結果とが最も相関関係が低い結果となった｡更に,冷水20℃浸水負荷30分後に測定して得られた結果を温水36℃浸水負荷10分後に測定して得られた結果で割った比を%で表示したところ回復率とこの比との間には正の相関関係(r=0.46,p<0.0001)が認められた｡このことから,温水36℃負荷時と,冷水20℃負荷時との差が大きい患者 において末梢皮膚温度の低下が著しいことが示された｡これまで悲観血的に測定されてきたサーモグラフィーによる末梢循環の数量的評価の試みは,レーザー血流計による結果と組み合わせることで,両者の間に正の相関関係が示されたことにより,今後,数量化された客観的評価を可能にした

サーモグラフィーによる体表面温度の測定 2.温水負荷の効果

The body surface temperature of 41 patients suffering coldness, numbness or pain in their feet was examined using thermography. Thermographic results were analyzed quantitatively by calculating a recovery ratio as: Recovery ratio =[Total counts of thermography (Pixels) over temperature (T) after cold loading] ÷ [Initial counts over T before cold loading] x 100(%). Three different baseline temperatures, 26℃. 27℃ and 28℃, were used in processing the thermographic results into pictures. The recovery ratio was susceptible to temperature, and we recommend a baseline temperature limitation of 27℃ for clinical study. A bi-modal distribution of recovery ratio was observed in 18 patients with diabetes mellitus. One group (6 subjects) had high recovery ratio between 80%-100%, and another group (10 subjects) had a low recovery ratio between 0%-19%. The results of thermography were also influenced by weather. To reduce the effect of outside temperature, we used pre-loading with hot water at 36℃ for 5 min (hot loading).　A large difference in　recovery ratio between presence and absence of hot loading was observed in 6 of the 30 subjects. The difference was over-estimated in more than 20% of recovery ratio without hot loading as compared with hot loading in these 6 subjects. The effect of drugs on peripheral circulation, such as beraprost sodium and sarpogrelate hydrochloride, was clear and quantified using thermography under these conditions of hot loading.下肢に冷感ならびにしびれ感または疼痛を訴える患者41症例についてサーモグラフィーを用いて体表面温度を測定した｡測定で得られた結果は回復率として数量化して表示された｡回復率の算出方法は回復率=[冷水負荷後の特定温度T℃以上の体表面温度のサーモグラフィーのPixelの総数]÷[温水負荷前の特定温度T℃以上の体表面温度のサーモグラフィーのPixelの総数]× 100%で求めた｡サーモグラフィーで得られた結果と画像処理の過程で用いられた,26℃,27℃,28℃の3つの異なる特定温度T℃ との関連について検討を行なった｡その結果,回復率は特定温度T℃に影響を受けやすいことが明らかとなった｡下肢の体表面温度の低い臨床症例においては27℃の条件が適当と考えられた｡前述の41症例中の18症例の糖尿病患者について検討を行なった｡そのサーモグラフィーの結果は,比較的回復率の高い(80%～100%)群の6症例と比較的回復率の低い(0%～19%)群の10症例の2群に別れた｡わずかに残り2症例が20%から79%の間であった｡下肢の症状が気温の低い時期に出親しやすいためにサーモグラフィーの検査を冬期に行なう必要性が高まった｡しかし,天候の影響を受けやすいために冷水負荷前の測定領域の下肢が冷えすぎているために20℃の室温に15分間の安静時間では体表面温度が十分に暖まることが出来ず,27℃以上の領域として測定範囲全体を観察できない 問題に直面した｡この間温点を解決する手段として36℃の温水に5分間下肢を入れて暖める温水負荷を加えることにした｡そこで, 温水負荷を行なった症例30症例について,温水負荷を行なう前(室温)の回復率と温水負荷を行なった後の回復率について比較検討を行なったところ,20%にあたる6症例において温水負荷を行なわなかった場合に20%以上の回復率の過剰評価が認めら れた｡温水負荷を行なうことにより年間を通じて天候の影響を最小限にすることが可能となり,この結果,長期間の内服薬の末梢循環に及ぼす影響の測 定を行なった場合に,季節の影響を最小限にしてサーモグラフィーにより回復率を用いて数値化された測定結果を検討することが可能となった｡具体的に末梢循環の改善に薬効が有ると言われている薬剤であるベラプロストおよびサルポグレラートを3ヵ月間内服した場合の前後のサーモグラフィーで得られた回復率について検討を行なった｡その結果はベラプロストにおいては,6.9%から41.9%に上昇または回復率の6.1倍の上昇を認めた。サルポグレラートにおいては,1.9%から17.3%に上昇または回復率の9.1倍の上昇を認めた。以上より,温水負荷を加えたサーモグラフィーの測定結果の数値化は下肢に症状の有る患者の末梢循環の評価ならびに薬効の評価の比較に有用であることが表わされた

気管支喘息患者における努力肺活量(% FVC)の低下と気道炎症ならびに臨床病型との関連

Clinical features of asthmatics with a low % FVC (<80%) were studied in relation to airway inflammation and clinical asthma types. 1. Twenty four (75.0%) of the 32 subjects with a low % FVC had steroid-dependent intractable asthma. 2. A significant association between % FVC value and clinical asthma type was found. The value of % FVC was significantly lower in subjects with type II asthma (bronchiolar obstruction), in which significantly decreased proportion of lymphocytes,and significantly increased proportion of neutrophils in bronchoalveolar lavage (BAL) fluid were observed, than in those with type la-1 (simple bronchoconstriction) with (P<0.001) and without glucocorticoid therapy (P<0.02). 3. The % FVC value was significantly improved after treatment in type lb (hypersecretion) with BAL eosinophilia, but not in those with type II with BAL neutrophilia. These results demonstrate that marked decrease of % FVC in patients with asthma correlated with airway inflammation, and that airway reversibility by medication is very low in patients with type II asthma.努力肺活量(% FVC)低値を示す気管支喘息の臨床病態を明らかにするために気道炎症ならびに臨床病型との関連について検討を行った｡% FVCが80%未満の気管支喘息患者32名を対象として,スパイロメトリー,気管支肺胞洗浄(BAL)を施行し,臨床病態を解析し,治療効果の評価を行なった｡1.対象中24名(75%)の患者がステロイド依存性重症難治性喘息であった｡2.細気管支閉塞型喘息患者において,% FVC値の有意の低下が認められ,BAL液中のリンパ球の減少と好中球の増加が認められた｡3.FVC値,FEV1.0億の治療による改善は,BAL液中好酸球の増加を示す過分泌型喘息においては認められが,BAL液中好中球の増加を示す細気管支閉塞型喘息においては認められなかった｡ 以上より喘息患者における% FVC値の低下は気道炎症と関連しており,細気管支閉塞型喘息における治療による可逆性は著しく低下していることが示唆された

中高年発症型喘息患者におけるIgE系アレルギー反応の加令による変化

Age-related changes in IgE-mediated allergic reaction were examined in patients with late onset asthma, whose asthma occured over the age at onset of 40. 1. The number of patients with a high serum IgE level over 3001U/mℓ was larger in those between the ages at onset of 40 and 49 and over age at onset 60 than in those between the ages at onset 50 and 59. 2. A positive RAST score to Hdm and cockroach allergens was more frequently found in patients between 40 and 49 and over age 60 at onset compared to those between 50 and 59 at onset. In contrast, the number of patients with a positive RAST to Candida was larger in those between 50 and 59 at onset than in those of between 40 and 49 and over 60 at onset. 3. Severe asthma was more frequently and slight asthma was less frequently observed in patients between 50 and 59 at onset than in those of the other age at onset groups. These results suggest that IgE-mediated allergic reaction in patients between the ages at onset of 50 and 59 is different from that in those with early onset asthma, and that the reaction in patients between the ages at onset 40 and 49 and over 60 resembles to the reaction in those with early onset asthma.発症年令が40才以上の中高年発症型喘息患者におけるIgE系アレルギー反応の加令による変化について検討を加えた｡1.発症年令が40から49才と60才以上の患者では,50から59才の患者に比して血清IgE値が300IU/mℓ以上の高値を示す頻度が高い傾向が見られた。2.ダニ,ゴキブリに対するRAST陽性率は発症年令が50から59才の患者に比して,40から49才と60才以上の患者で高値を示した｡一方,カンジダに対するRAST陽性率は,発症年令が40から49才と60才以上の患者に比して,50から59才の患者で高値を示した｡3.発症年令が50から59才の患者では,他の発症年令の患者に比べて,重症例が多く,軽症例が少ない傾向が見られた｡以上の結果から,発症年令が50から59才の患者のIgE系アレルギー反応は若年発症型喘息患者と異なり,発症年令が40から49才と60才以上の患者のIgE系アレルギー反応は若年発症型喘息患者と 類似することが示唆された

Establishment of a novel collagenase perfusion method to isolate rat pancreatic stellate cells and investigation of their gene expression of TGF-beta1, type I collagen, and CTGF in primary culture or freshly isolated cells.

In studies of the pathogenesis of pancreatic fibrosis, pancreatic stellate cells (PSCs) have recently gained attention. In the present study, we established a new collagenase perfusion method through thoracic aorta cannulation to isolate PSCs, and we studied gene expression of TGF-beta1, type I collagen, and connective tissue growth factor using primary cultured PSCs. Our method facilitated PSC isolation, and by our new method, 4.3 +/- 1.2 x 10(6) PSCs were obtained from a rat. In comparing the expression of these genes with that of hepatic stellate cells (HSCs), we observed a similar pattern, although PSCs expressed type I collagen gene earlier than did HSCs. These results suggest that PSCs may play an important role in fibrosis of the pancreas, as HSCs do in liver fibrosis; in addition, PSCs may exist in a preactivated state or may be more easily activated than are HSCs. We also isolated the PSCs from a WBN/Kob rat, the spontaneous pancreatitis rat, and compared the gene expression with that from a normal rat

気管支喘息におけるアレルギー性鼻炎と花粉抗原に対するIgE抗体

IgE antibodies against inhalant allergens were estimated in 53 patients with bronchial asthma in relation to allergic rhinitis. Of them, 20 patients (37.7%) had allergic rhinitis. 1. Asthma + allergic rhinitis were often observed in patients between the ages of 0 and 39. In contrast, asthma alone in those over age 60. 2. A RAST score was positive in house dust mite (HDm)(50.9%), cockroach(24.5%), and Candida (26.4%) in these patients, and the positive rate was not different between patients with and without allergic rhinitis. 3. The frequency of positive RAST against Japanese cedar and rice plants was higher in patients with allergic rhinitis (42.9% and 18.5%) than in those without allergic rhinitis (28.6 and 3.7%), however, the each positive rate or RAST against the two allergens was not significantly different between those with AR and without AR. 4. The number of patients with AR induced by pollen alone (pollinosis) was not large ( 5/53, 9.4%) in the patients with asthma. It was clarified from the results that allergic rhinitis was often observed in asthma patients, but not pollinosis. and that IgE antibodies against Japanese cedar and rice plants were found even in patients without AR.気管支喘息53例を対象に,吸入抗原に対するIgE抗体とアレルギー性鼻炎の合併の有無との関連について若干の検討を加えた｡対象53例中20例(37.7%)にアレルギー性鼻炎の合併が見られた｡ 1.喘息+アレルギー性鼻炎の合併は,0-39才の年齢層で最も多い傾向であったが,一方,喘息単独は60才以上の症例に多く見られた｡2,IgE抗体の陽性率は,それぞれHDm50.9%,ごきぶり 24.5%,カンジダ26.4%であった｡これら抗原のRAST陽性率は,アレルギー性鼻炎合併例と非合併例の問に有意の差は見られなかった｡3.スギおよびイネ科の花粉に対するRAST陽性率はアレルギー性鼻炎合併例で(スギ42.9%,イネ科18.5%),非合併例に比べ(28.6%と3.7%)高い傾向が見られたが,両群間に有意の差は見られなかった｡4.花粉抗原によるアレルギー性鼻炎(花粉症)の頻度は9.4% (53例中5列)とあまり高くはなかった｡以上の結果より,気管支喘息患者でしばしばアレルギー性鼻炎の合併が見られること,(しかし, 花粉症は少ない),そして,スギおよびイネ科の花粉に対するIgE抗体は,アレルギー性鼻炎の合併のない症例においても観察されること,などが明らかにされた

気管支喘息に対する複合温泉療法の作用機序 1.温泉療法の評価方法との関連

Action mechanisms of complex spa therapy (swimming training in a hot spring pool, inhalation with iodine salt solution, and fango therapy) were studied in relation to patient age, clinical asthma type, and airway inflammation. Actions of spa therapy were observed by two clinical effects : direct and indirect effects. Improvement of subjective symptoms, ventilatory function, and bronchial hyperresponsiveness was observed as direct effects of spa therapy, and improvement of suppressed function of adrenocortical glands as indirect effects of the therapy. The clinical effects of spa therapy were higher in patients over the age of 40. Regarding clinical asthma type, the effects were larger in patients with type Ib and type II asthma than in those with type Ia. The efficacy of spa therapy was closely related with airway inflammation in patients with type Ib and type II asthma.複合温泉療法(温泉プール水泳訓練,ヨードゾル吸入,鉱泥湿布療法)の作用機序が,年齢,臨床病型,気道炎症反応との関連のもとに検討された｡温泉療法の作用機序は,2種類の臨床効果,すなわち,直接効果と間接効果により観察された｡自,他覚症状の改善,換気機能の改善,そして,気道過敏性の改善などが,温泉療法の直接効果として,また,副腎皮質機能の改善が間接効果とし て観察された｡温泉療法の臨床効果は,40才以上の症例においてより有効であり,また,臨床病型別では､Ia型に比べ,Ib型およびⅡ型においてより有効であった｡Ib型やⅡ型の気管支喘息では,温泉療法の作用機序と気道炎症反応との間に密接な関連が認められた