Design of engineered active zymogen of microbial transglutaminase

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当院の子宮下部筋腫合併妊娠における分娩転帰について

　子宮筋腫は年齢と共に有病率が増加する。それ故、近年における晩婚化や出産年齢の高齢化に伴い、子宮筋腫合併妊娠も増加傾向にある。比較的大きな子宮筋腫が子宮下部にある場合、経腟分娩困難と判断され帝王切開が選択される症例も少なくないと考えられる。　子宮筋腫の位置及び大きさと経腟分娩の可否について検討するため、当院の外来にて経腟分娩困難となる可能性があると判断された子宮筋腫合併妊娠で、妊娠後期に核磁気共鳴画像法(Magnetic Resonance Imaging:以下MRI)が撮影された6症例に対して後方視的に検討した。6症例中5症例で経腟分娩が可能であったが、子宮体下部後壁に長径90mmの子宮筋腫を認めた1例は、妊娠41週で予定日超過のため誘発を開始したが、分娩停止のため帝王切開による分娩となった。帝王切開の1例を他の5例と比較すると、年齢、妊娠週数、子宮筋腫の大きさに特記すべき差異を認めないが、子宮筋腫の位置がほぼ正中で、かつ子宮筋腫の下端が内子宮口よりも低位であるとの特徴を認めた。逆に、長径70〜120mmの子宮下部筋腫が存在してもその位置が正中から偏心している場合や、位置が正中でも下端が内子宮口よりも高位であれば、経腟分娩が可能であった

生体分子高感度検出を目指した酵素融合タンパク質試薬の開発

主1工学_化学システム工

Predictive factors of labour onset using ultrasonography

We analysed the effectiveness of transvaginal ultrasonographic and foetal/maternal pulse Doppler findings as predictors of labour onset within 1 week. We included 22 single normal pregnancies and evaluated the one-point and short- and long-term differences in uterine artery pulsatility index (PI), umbilical artery PI, middle cerebral artery PI (MCA-PI), peak systolic velocity, and cervical length (CL). Presence of funnelling and membrane separation over the internal cervical os was evaluated. Significant changes were observed in the one-point measurement of and short-term and long-term differences in CL, the one-point measurement of and long-term difference in MCA-PI, and the presence of membrane separation (Grade 2). In multivariate analysis, the significant predictors were short-term differences in CL (odds ratio [OR]: 5.27), long-term differences in MCA-PI (OR: 13.3), and presence of membrane separation (Grade 2) (OR: 5.38). Transvaginal ultrasonographic and foetal pulse Doppler findings were effective predictors of labour onset within 1 week.Impact statement What is already known on this subject? Parameters reported to predict labour onset include the Bishop score, cervical length, decreased long-term cervical length, funnelling of the internal cervical os, and adrenal gland volume. What do the results of this study add? Short-term changes in cervical length, long-term changes in middle cerebral artery pulsatility index, and the presence of membrane separation Grade 2 were found to be useful predictive factors of labour onset in this study. What are the implications of these findings for clinical practice and/or further research? The prediction of labour onset enables clinicians to properly manage pregnancy and delivery considering maternal and foetal conditions

Protein-Grafted Polymers Prepared Through a Site-Specific Conjugation by Microbial Transglutaminase for an Immunosorbent Assay

Protein–polymer conjugates have been developed in many fields. Most hybrids are composed of one protein attached to one or several polymer chains. The other form of hybrid involves the construction of multiple proteins on one polymer chain, thereby facilitating protein assemblies that provide multivalent effects. Unfortunately, synthetic methods for production of these types of hybrids are limited and challenging because precise control of the conjugation sites is needed. Herein, a novel synthetic polymer that can enzymatically assemble multiple proteins was developed. Polyacrylamide grafted with multiple microbial transglutaminase (MTG)-recognizable peptide derivatives was synthesized, and MTG-catalyzed site-specific conjugation of proteins with the polymer was achieved. The application for immunological biosensing was demonstrated using the assembly of a fusion protein composed of antibody-binding and enzyme moieties. This enzymatic method to synthesize a one-dimensional protein assembly on a synthetic polymer is versatile and can be expanded to a wide range of applications

Circadian Clock Proteins LHY and CCA1 Regulate SVP Protein Accumulation to Control Flowering in Arabidopsis[W]

The floral regulators GIGANTEA (GI), CONSTANS (CO), and FLOWERING LOCUS T (FT) play key roles in the photoperiodic flowering responses of the long-day plant Arabidopsis thaliana. The GI-CO-FT pathway is highly conserved in plants. Here, we demonstrate that the circadian clock proteins LATE ELONGATED HYPOCOTYL (LHY) and CIRCADIAN CLOCK–ASSOCIATED1 (CCA1) not only repressed the floral transition under short-day and long-day conditions but also accelerated flowering when the plants were grown under continuous light (LL). LHY and CCA1 accelerated flowering in LL by promoting FT expression through a genetic pathway that appears to be independent of the canonical photoperiodic pathway involving GI and CO proteins. A genetic screen revealed that the late-flowering phenotype of the lhy;cca1 double mutant under LL was suppressed through mutations in SHORT VEGETATIVE PHASE (SVP), a MADS box transcription factor. Yeast two-hybrid analysis demonstrated an interaction between SVP and FLOWERING LOCUS C, and genetic analysis indicated that these two proteins act as partially redundant repressors of flowering time. SVP protein accumulated in lhy;cca1 plants under LL. We propose a model in which LHY and CCA1 accelerate flowering in part by reducing the abundance of SVP and thereby antagonizing its capacity to repress FT expression under LL

WNT7A regulates tumor growth and progression in ovarian cancer through the WNT/β-catenin pathway

Abnormal activation the WNT/β-catenin signaling pathway has been associated with ovarian carcinomas, but a specific WNT ligand and pertinent downstream mechanisms are not fully understood. In this study, we found abundant WNT7A in the epithelium of serous ovarian carcinomas, but not detected in borderline and benign tumors, normal ovary or endometrioid carcinomas. To characterize the role of WNT7A in ovarian tumor growth and progression, nude mice were injected either intraperitoneally (i.p.) or subcutaneously (s.c.) with WNT7A knocked down SKOV3.ip1 and overexpressed SKOV3 cells. In the i.p. group, mice receiving SKOV3.ip1 cells with reduced WNT7A expression developed significantly fewer tumor lesions. Gross and histological examination revealed greatly reduced invasion of WNT7A knockdown cells into intestinal mesentery and serosa compared to the control cells. Tumor growth was regulated by loss or overexpression of WNT7A in mice receiving s.c. injection as well. In vitro analysis of cell function revealed that cell proliferation, adhesion, and invasion were regulated by WNT7A. The activity of the TCF/LEF reporter was stimulated by overexpression of WNT7A in ovarian cancer cells. Co-transfection with WNT7A and FZD5 receptor further increased activity, and this effect was inhibited by co-transfection with SFRP2, or dominant-negative TCF4. Overexpression of WNT7A stimulated MMP7 promoter, and mutation of TCF binding sites in MMP7 promoter confirmed that activation of MMP7 promoter by WNT7A was mediated by β-catenin/TCF signaling. Collectively, these results suggest that re-expression of WNT7A during malignant transformation of ovarian epithelial cells plays a critical role in ovarian cancer progression mediated by WNT/β-catenin signaling pathway