NMDA-Receptor Activation Induces Calpain-Mediated β-Catenin Cleavages for Triggering Gene Expression

SummaryThe canonical Wnt-β-catenin signaling pathway is important for a variety of developmental phenomena as well as for carcinogenesis. Here, we show that, in hippocampal neurons, NMDA-receptor-dependent activation of calpain induced the cleavage of β-catenin at the N terminus, generating stable, truncated forms. These β-catenin fragments accumulated in the nucleus and induced Tcf/Lef-dependent gene transcription. We identified Fosl1, one of the immediate-early genes, as a target of this signaling pathway. In addition, exploratory behavior by mice resulted in a similar cleavage of β-catenin, as well as activation of the Tcf signaling pathway, in hippocampal neurons. Both β-catenin cleavage and Tcf-dependent gene transcription were suppressed by calpain inhibitors. These findings reveal another pathway for β-catenin-dependent signaling, in addition to the canonical Wnt-β-catenin pathway, and suggest that this other pathway could play an important role in activity-dependent gene expression

プロテアソーム阻害剤エポキソマイシンとMG262はヒト口腔扁平上皮癌細胞(HSC-3)によるウロキナーゼ型プラスミノーゲンアクチベータ発現を抑制する

プロテアソーム阻害剤のエポキソマイシンとMG262が、HSC-3細胞におけるウロキナーゼ型プラスミノーゲンアクチベータ(uPA)のmRNA発現とuPA産生に及ぼす影響について検討した。エポキソマイシンとMG262はuPA mRNA発現とuPA産生を用量依存性に抑制した。経時変化の検討では、エポキソマイシンまたはMG262への曝露から6時間程度で、uPA mRNA発現に顕著な減少が認められ、抑制効果は12および24時間後と同等またはやや強かった。エポキソマイシンとMG262は核性因子(NF)-κB-依存性プロモーターの転写活性を弱めた。uPA遺伝子の転写はNF-κB活性に依存することが知られており、これらのプロテアソーム阻害剤によるuPA遺伝子発現の抑制は、構成的NF-κB活性阻害の影響と考えられる。またエポキソマイシンとPG262はHSC-3細胞の浸潤活性を低下させた。プロテアソーム阻害剤による浸潤活性の減少はuPA産生抑制の影響を一部に受けていると考えられた

Purification of rabbit tumor necrosis factor

AbstractRabbit tumor necrosis factor (TNF) was purified and shown by SDS-PAGE to be a single protein of 18 kDa. TNF in 355 ml rabbit serum was precipitated with ammonium sulfate, and purified by repeated DEAE-Sephadex and Sephacryl S-200 chromatographies, and the final fractionation on Blue-Sepharose 6B. By this procedure its yield was 22% and its specific activity was 2.4 × 107 U/mg protein. The sequence of the N-terminal 20 amino acids was determined

Test of mp/me changes using vibrational transitions in N2+

In this paper we propose to utilize the X2Σg(ν,N,F,M)=(0,0,1/2,±1/2)→(1,0,1/2,±1/2) or (2,0,1/2,±1/2) transition of N2+ (I=0) to test variations of the proton-to-electron mass ratio. The X2Σg ground state exhibits no quadrupole shift and the Zeeman shift of the N=0→N=0 transition is exactly zero. Because N2+ is nonpolar, systematic level shifts such as Stark shifts induced by trap electric field or blackbody radiation are very small and the thermalization of the rotational states is inhibited. This eases the requirements on the experimental setup significantly. Employing Raman transitions at the “magic” wavelength the (0,0,1/2,±1/2)→(1,0,1/2,±1/2) or (2,0,1/2,±1/2) transition frequency can be measured very precisely

Detection of miRNA in Cell Cultures by Using Microchip Electrophoresis with a Fluorescence-Labeled Riboprobe

The analysis of a microRNA (miRNA), miR-222 isolated from the PC12 cell line, was performed by use of the ribonuclease (RNase) protection assay, cyanine 5 (Cy5)-labeled miR-222 riboprobe, and a Hitachi SV1210 microchip electrophoresis system, which can be used to evaluate the integrity of total RNA. The fluorescence intensity corresponding to the protected RNA fragment increased in a dose-dependent manner with respect to the complementary-strand RNA. More highly sensitive detection of miRNA by microchip electrophoresis than by conventional method using fluorescence-labeled riboprobe could be obtained in 180 s. An obvious increase in miR-222 expression induced by nerve growth factor in PC12 cells could be observed. These results clearly indicate the potential of microchip electrophoresis for the analysis of miRNA using RNase protection assay with a fluorescence-labeled riboprobe

Ulinastatin did not reduce mortality in elderly multiple organ failure patients: a retrospective observational study in a single center ICU

AimOur aim was to evaluate the effect of ulinastatin on 28‐day mortality in patients who developed multiple organ failure (MOF) related to their acute illness and were admitted to the intensive care unit (ICU).MethodsWe carried out a retrospective observational study of MOF patients in a general ICU of a tertiary care hospital in Japan from January 2009 to December 2012. The primary outcome was 28‐day all‐cause mortality. Secondary outcomes were ventilator‐free days, ICU‐free days, and vasopressor‐free days at day 28. We investigated the association between ulinastatin treatment and outcomes using multivariable regression analysis.ResultsA total of 212 MOF patients were included, 79 (37%) of whom received ulinastatin. The median age was 70 years (interquartile range, 60–77) and median APACHE II score was 25 (interquartile range, 19–29). Overall 28‐day mortality was 20%. There were no significant differences between the ulinastatin group and the control group in age, gender, or APACHE II score. The ulinastatin group had higher prevalence of sepsis (44% versus 22%, P = 0.001). Multivariable logistic regression analysis showed that ulinastatin was not associated with 28‐day mortality (odds ratio = 1.22; 95% confidence interval, 0.54–2.79). Moreover, ulinastatin did not reduce the mortality in patients with sepsis (odds ratio = 1.92; 95% confidence interval, 0.52–7.13). However, ICU‐free days and ventilator‐free days was significantly fewer in the ulinastatin group than control group.ConclusionsIn this retrospective observational study, ulinastatin was not associated with mortality in elderly patients with established MOF, although it might be related to patient\u27s utility

Transient improvement of urticaria induces poor adherence as assessed by Morisky Medication Adherence Scale-8.

Poor adherence to medication is a major public health challenge. Here, we aimed to determine the adherence to oral and topical medications and to analyze underlying associated factors using the translated Japanese version of Morisky Medication Adherence Scale-8 regarding urticaria treatment. Web-based questionnaires were performed for 3096 registered dermatological patients, along with a subanalysis of 751 registered urticaria patients in this study. The adherence to oral medication was significantly associated with the frequency of hospital visits. Variables that affected the adherence to topical medication included age and experience of drug effectiveness. The rate of responses that "It felt like the symptoms had improved" varied significantly among the dermatological diseases treated with oral medications. Dermatologists should be aware that adherence to the treatment of urticaria is quite low. Regular visits and active education for patients with urticaria are mandatory in order to achieve a good therapeutic outcome by increasing the adherence