Comparison of recovery of mobility and self-efficacy after total knee arthroplasty based on two different protocols: A prospective cohort study

Objectives: The purpose of this study was to compare the recovery of mobility and self-efficacy following total knee arthroplasty (TKA) between the 5-day and the 28-day protocol. This prospective cohort study was carried out at two hospitals. Methods: In total, 104 patients who underwent TKA were enrolled. The primary outcomes measured were Life Space Assessment (LSA) for mobility and modified-Gait Efficacy Scale (mGES) for self-efficacy. Knee Society Score (KSS) was used to estimate the functional outcomes. These assessments were performed in all patients preoperatively, and at 1, 3, and 6 months postoperatively. After calculating the propensity score using covariates, such as patient characteristics, LSA, mGES, and KSS at baseline, propensity score-adjusted multivariate analysis of covariance (MANCOVA) was performed. Results: MANCOVA revealed significant differences in LSA and mGES, but not in KSS, between the two protocols. The adjusted means of LSA and mGES in the 28-day protocol were significantly greater than those in the 5-day protocol in all the postoperative assessments. Conclusion: Mobility and self-efficacy were greater following the 28-day protocol than the 5-day protocol after TKA. Our findings suggest that the modified treatment procedure for improving mobility and self-efficacy is necessary to introduce the early discharge protocol in Japan

Amino acids C-terminal to the 14-3-3 binding motif in CDC25B affect the efficiency of 14-3-3 binding

金沢大学医薬保健研究域薬学系The phospho-site adapter protein 14-3-3 binds to target proteins at amino acid sequences matching the consensus motif Arg-X-X-Ser/Thr-X-Pro, where the serine or threonine residue is phosphorylated and X is any amino acid. The dual-specificity phosphatase CDC25B, which is involved in cell cycle regulation, contains five 14-3-3 binding motifs, but 14-3-3 preferentially binds to the motif at Ser309 in CDC25B1 (or Ser323 in CDC25B3). In the present study, we demonstrate that amino acid residues C-terminal to the 14-3-3 binding motif strongly affect the efficiency of 14-3-3 binding. Alanine substitutions at residues downstream of the Ser309 motif dramatically reduced 14-3-3 binding, although phosphorylation of Ser309 was unaffected. We also observed that binding of endogenous 14-3-3 to mutant CDC25B occurred less efficiently than to the wild type. Mutants to which 14-3-3 cannot bind efficiently tend to be located in the nucleus, although not as specifically as the alanine substitution mutant of Ser309. These results indicate that amino acid sequences C-terminal to the consensus binding site have an important role in the efficient binding of 14-3-3 to at least CDC25B, which may partly explain why some consensus sequences are inactive as 14-3-3 binding sites. © 2006 The Japanese Biochemical Society

Androgen-independent proliferation of LNCaP prostate cancer cells infected by xenotropic murine leukemia virus-related virus

Xenotropic murine leukemia virus-related virus (XMRV) is a novel gammaretrovirus that was originally isolated from human prostate cancer. It is now believed that XMRV is not the etiologic agent of prostate cancer. An analysis of murine leukemia virus (MLV) infection in various human cell lines revealed that prostate cancer cell lines are preferentially infected by XMRV, and this suggested that XMRV infection may confer some sort of growth advantage to prostate cancer cell lines. To examine this hypothesis, androgen-dependent LNCaP cells were infected with XMRV and tested for changes in certain cell growth properties. We found that XMRV-infected LNCaP cells can proliferate in the absence of the androgen dihydrotestosterone. Moreover, androgen receptor expression is significantly reduced in XMRV-infected LNCaP cells. Such alterations were not observed in uninfected and amphotropic MLV-infected LNCaP cells. This finding explains why prostate cancer cell lines are preferentially infected with XMRV

Protein kinase Cδ binds TIRAP/Mal to participate in TLR signaling

Toll-like receptor (TLR) family members recognize specific molecular patterns within pathogens. Signaling through TLRs results in a proximal event that involves direct binding of adaptor proteins to the receptors. We observed that TIRAP/Mal, an adaptor protein for TLR2 and TLR4, binds protein kinase Cδ (PKCδ). TIRAP/Mal GST-fusion protein and a TIRAP/Mal antibody were able to precipitate PKCδ from rat peritoneal macrophage and THP1 cell lysates. Truncation mutants of TIRAP/Mal showed that the TIR domain of TIRAP/Mal is responsible for binding. TLR2- and TLR4-mediated phosphorylation of p38 MAPK, IKK, and IκB in RAW264.7 cells were abolished by depletion of PKCδ. These results suggest that PKCδ binding to TIRAP/Mal promotes TLR signaling events

Infection of XC Cells by MLVs and Ebola Virus Is Endosome-Dependent but Acidification-Independent

Inhibitors of endosome acidification or cathepsin proteases attenuated infections mediated by envelope proteins of xenotropic murine leukemia virus-related virus (XMRV) and Ebola virus, as well as ecotropic, amphotropic, polytropic, and xenotropic murine leukemia viruses (MLVs), indicating that infections by these viruses occur through acidic endosomes and require cathepsin proteases in the susceptible cells such as TE671 cells. However, as previously shown, the endosome acidification inhibitors did not inhibit these viral infections in XC cells. It is generally accepted that the ecotropic MLV infection in XC cells occurs at the plasma membrane. Because cathepsin proteases are activated by low pH in acidic endosomes, the acidification inhibitors may inhibit the viral infections by suppressing cathepsin protease activation. The acidification inhibitors attenuated the activities of cathepsin proteases B and L in TE671 cells, but not in XC cells. Processing of cathepsin protease L was suppressed by the acidification inhibitor in NIH3T3 cells, but again not in XC cells. These results indicate that cathepsin proteases are activated without endosome acidification in XC cells. Treatment with an endocytosis inhibitor or knockdown of dynamin 2 expression by siRNAs suppressed MLV infections in all examined cells including XC cells. Furthermore, endosomal cathepsin proteases were required for these viral infections in XC cells as other susceptible cells. These results suggest that infections of XC cells by the MLVs and Ebola virus occur through endosomes and pH-independent cathepsin activation induces pH-independent infection in XC cells

Susceptibility of muridae cell lines to ecotropic murine leukemia virus and the cationic amino acid transporter 1 viral receptor sequences: implications for evolution of the viral receptor

Ecotropic murine leukemia viruses (Eco-MLVs) infect mouse and rat, but not other mammalian cells, and gain access for infection through binding the cationic amino acid transporter 1 (CAT1). Glycosylation of the rat and hamster CAT1s inhibits Eco-MLV infection, and treatment of rat and hamster cells with a glycosylation inhibitor, tunicamycin, enhances Eco-MLV infection. Although the mouse CAT1 is also glycosylated, it does not inhibit Eco-MLV infection. Comparison of amino acid sequences between the rat and mouse CAT1s shows amino acid insertions in the rat protein near the Eco-MLV-binding motif. In addition to the insertion present in the rat CAT1, the hamster CAT1 has additional amino acid insertions. In contrast, tunicamycin treatment of mink and human cells does not elevate the infection, because their CAT1s do not have the Eco-MLV-binding motif. To define the evolutionary pathway of the Eco-MLV receptor, we analyzed CAT1 sequences and susceptibility to Eco-MLV infection of other several murinae animals, including the southern vole (Microtus rossiaemeridionalis), large Japanese field mouse (Apodemus speciosus), and Eurasian harvest mouse ( Micromys minutus). Eco-MLV infection was enhanced by tunicamycin in these cells, and their CAT1 sequences have the insertions like the hamster CAT1. Phylogenetic analysis of mammalian CAT1s suggested that the ancestral CAT1 does not have the Eco-MLV-binding motif, like the human CAT1, and the mouse CAT1 is thought to be generated by the amino acid deletions in the third extracellular loop of CAT1

Time-resolved serial femtosecond crystallography reveals early structural changes in channelrhodopsin

X線自由電子レーザーを用いて、光照射によるチャネルロドプシンの構造変化の過程を捉えることに成功. 京都大学プレスリリース. 2021-03-26.Channelrhodopsins (ChRs) are microbial light-gated ion channels utilized in optogenetics to control neural activity with light . Light absorption causes retinal chromophore isomerization and subsequent protein conformational changes visualized as optically distinguished intermediates, coupled with channel opening and closing. However, the detailed molecular events underlying channel gating remain unknown. We performed time-resolved serial femtosecond crystallographic analyses of ChR by using an X-ray free electron laser, which revealed conformational changes following photoactivation. The isomerized retinal adopts a twisted conformation and shifts toward the putative internal proton donor residues, consequently inducing an outward shift of TM3, as well as a local deformation in TM7. These early conformational changes in the pore-forming helices should be the triggers that lead to opening of the ion conducting pore

遠隔教育の実施と大学での教育に関する一考察 ―建学の精神を伝える授業のオンラインでの実施をもとに―

2019年末に端を発した新型コロナ感染症（COVID-19）の拡大によって、多くの大学は2020年度の新学期から遠隔教育の実施に踏み切った。本学においても遠隔教育の採用が決定され、ホスピタリティ概論も IT 機器を活用したオンラインでの実施となった。この中で大学教育の将来像に大きな影響を与えると考えられるオンライン授業の在り方を探る基礎調査を実施した。その結果、次のことが明らかになった。1）受講生の IT 環境の整備には、差がみられること、2）受講生の使用する機器は、パソコン、タブレット、スマートフォンに分かれること、3）オンライン授業に関しては、肯定する意見がある一方、改善を要望する意見も見られること、4）スマートフォンのみでの受講者とパソコンおよびタブレットでの受講者を比較分析した結果、画面の明瞭度や授業の進行、及び授業の理解などにおいて両者に差がみられ、スマートフォンのみでの受講者の方が有意に低い結果であったこと、5）遠隔授業に関する要望等の中には、遠隔教育の利点を述べている受講生の他に、授業以外の大学が持つ機能、つまり、友人獲得や相互啓発に関する不安も多くみられたこと。この結果等を踏まえ、オンライン授業のあり方、及び、将来社会における大学教育のあり方に関する提案を行った