Simultaneous monitoring of independent gene expression patterns in two types of cocultured fibroblasts with different color-emitting luciferases

<p>Abstract</p> <p>Background</p> <p>Luciferase assay systems enable the real-time monitoring of gene expression in living cells. We have developed a dual-color luciferase assay system in which the expression of multiple genes can be tracked simultaneously using green- and red-emitting beetle luciferases. We have applied the system to monitoring independent gene expressions in two types of cocultured fibroblasts in real time.</p> <p>Results</p> <p>Two Rat-1 cell lines were established that stably express either green- or red-emitting luciferases under the control of the <it>mBmal1 </it>promoter, a canonical clock gene. We cocultured these cell lines, and gene expression profiles in both were monitored simultaneously. The circadian rhythms of these cell lines are independent, oscillating following their intrinsic circadian phases, even when cocultured. Furthermore, the independent rhythms were synchronized by medium change as an external stimulus.</p> <p>Conclusion</p> <p>Using this system, we successfully monitored independent gene expression patterns in two lines of cocultured fibroblasts.</p

Hypotaurine is an Energy-Saving Hepatoprotective Compound against Ischemia-Reperfusion Injury of the Rat Liver

Metabolome analyses assisted by capillary electrophoresis-mass spectrometry (CE-MS) have allowed us to systematically grasp changes in small molecular metabolites under disease conditions. We applied CE-MS to mine out biomarkers in hepatic ischemia-reperfusion. Rat livers were exposed to ischemia by clamping of the portal inlet followed by reperfusion. Metabolomic profiling revealed that l contents of taurine in liver and plasma were significantly increased. Of interest is an elevation of hypotaurine, collectively suggesting significance of hypotaurine/taurine in post-ischemic responses. Considering the anti-oxidative capacity of hypotaurine, we examined if supplementation of the compound or its precursor amino acids could affect hepatocellular viability and contents of taurine in liver and plasma. Administration of hypotaurine, N-acetylcysteine or methionine upon reperfusion comparablly attenuated the post-ischemic hepatocellular injury but with different metabolomic profiling among groups: rats treated with methionine or N-acetylcysteine but not those treated with hypotaurine, exhibited significant elevation of hepatic lactate generation without notable recovery of the energy charge. Furthermore, the group treated with hypotaurine exhibited elevation of the plasma taurine, suggesting that the exogenously administered compound was utilized as an antioxidant. These results suggest that taurine serves as a surrogate marker for ischemia-reperfusion indicating effectiveness of hypotaurine as an energy-saving hepatoprotective amino acid

Association between Public Assistance and Frequent Emergency Department Visits in Urban Areas of Japan: A Case-Control Study

Emergency department（ED）crowding due to frequent visitors increases medical costs and endangers the safety of patients who need prompt treatment. Although some studies have examined the social background of frequent visitors to the ED, there are few comprehensive studies of factors related to frequent ED visits in Japan. The present study examined whether public assistance status, age, and underlying conditions are associated with frequent visits to the ED. Subjects in this study were patients over 20 years of age who visited the ED of General Medicine in Showa University Hospital in Tokyo, Japan, from April 1, 2014, to March 31, 2015（n＝6,182）. Information regarding sex, age, public insurance, hypertension, diabetes mellitus, dyslipidemia, chronic obstructive pulmonary disease, cancer, dysuria, bronchial asthma, and chronic kidney disease was collected using a patient database created from medical and hospital records. Based on a previous study, subjects who visited the ED four or more times were defined as frequent visitors, while those who visited the ED less than four times were considered infrequent visitors. Logistic regression analysis was performed to calculate odds ratios（ORs）and 95％ confidence intervals（CIs）for frequent ED visits. Receipt of public assistance significantly increased the OR for frequent visits（4.26, 95％ CI: 1.90-9.56）, even after adjusting for sex, age, and potential confounding factors（OR: 3.89, 95％ CI: 1.62-9.35）. The current study found a significant association between frequent ED visits and the receipt of public assistance. Emergency physicians and medical practitioners need to share information with outpatient physicians and welfare officers and work together with patients to address their specific medical and social vulnerabilities

Acute exercise induces biphasic increase in respiratory mRNA in skeletal muscle

Peroxisome proliferator-activated receptor c coactivator-1α (PGC-1α) promotes the expression of oxidative enzymes in skeletalmuscle. We hypothesized that activation of the p38 MAPK (mitogen-activated protein kinase) in response to exercise was associatedwith exercise-induced PGC-1α and respiratory enzymes expression and aimed to demonstrate this under the physiological level. Wesubjected mice to a single bout of treadmill running and found that the exercise induced a biphasic increase in the expression ofrespiratory enzymes mRNA. The second phase of the increase was accompanied by an increase in PGC-1α protein, but the otherwas not. Administration of SB203580 (SB), an inhibitor of p38 MAPK, suppressed the increase in PGC-1α expression and respiratoryenzymes mRNA in both phases. These data suggest that p38 MAPK is associated with the exercise-induced expression of PGC-1α andbiphasic increase in respiratory enzyme mRNAs in mouse skeletal muscle under physiological conditions

小学生向け外来種＆ヒアリ学習ワークショップの開発と実践

市民のヒアリ監視活動への参画，および外来種リテラシーの向上を目指した体験型ワークショップ（以下，WS）を開発した．ヒアリの日本国土への定着を阻止するためには，専門家だけでなく市民ひとりひとりがヒアリ監視のスキルと意識を持つことが，きわめて有効なリスク対策となる．本WS は，外来種問題という社会が抱える喫緊課題に対する問題解決の手段としての側面と，市民が身近な自然の生物多様性を学ぶ環境学習の側面を併せ持つ．そのため，ヒアリや外来種に対する危機意識の醸成だけではなく，生き物への純粋な興味や知識欲をくすぐるエンターテイメント性を意識してプログラムをデザインした．加えて，生物を扱うWS で最も講師のスキルと経験が求められるパートである野外観察・採集と顕微鏡観察を省略するために，アリ類の精密拡大模型を作成した．これにより，専門性を担保しつつも，専門家でなくとも実施可能な比較的手軽で汎用性の高いWSとなった．WS のコンパクト化を実現したことで，危機管理WS の命題である実施範囲の拡大へとつながった．ヒアリ対策のニーズの高い沖縄県において継続的にWS を実践し，改良を重ねて本WS が完成した．WS の前後でとった参加者へのアンケート調査結果から，本WS の最適な実施対象は小学校中学年であること，参加者の「ヒアリ」のキーワード認識率はWS 前から高い水準にあること，そしてWS 参加によって「ヒアリ」および「外来種」のキーワード認識率が上昇することが明らかになった