Expression of a Chitinase Gene and Lysis of the Host Cell Wall during Chlorella Virus CVK2 Infection

AbstractA chitinase gene (vChti-1) encoded by the Chlorella virus CVK2 was cloned and characterized. The vChti-1 open reading frame consisted of 2508 bp corresponding to 836 amino acid residues. The predicted amino acid sequence contained two sets of a family 18 catalytic domain that is responsible for chitinase activity. Northern blot analysis revealed that the vChti-1 gene was expressed in virus-infected Chlorella cells late in infection, when a single transcript of about 2.5 kb appeared at 120 min postinfection. This result was confirmed by Western blotting with a specific anti-vChti-1 protein antibody; a protein of about 94 kDa was detected specifically beginning at 240 min postinfection and was present until cell lysis. The protein was not incorporated into viral particles but remained in the medium after cell lysis. The vChti-1 protein produced in virus-infected cells showed chitinase activity on zymogram assays

Oxidative modification of IκB by monochloramine inhibits tumor necrosis factor α-induced NF-κB activation

AbstractWe have previously reported that monochloramine (NH2Cl), a neutrophil-derived oxidant, inhibited tumor necrosis factor α (TNFα)-induced expression of cell adhesion molecules and nuclear factor-κB (NF-κB) activation (Free Radical Research 36 (2002) 845–852). Here, we studied the mechanism how NH2Cl inhibited TNFα-induced NF-κB activation, and compared the effects with taurine chloramine (Tau–NHCl). Pretreatment of Jurkat cells with NH2Cl at 70 μM resulted in suppression of TNFα-induced IκB phosphorylation and degradation, and inhibited NF-κB activation. In addition, a slow-moving IκB band appeared on SDS-PAGE. By contrast, Tau–NHCl for up to 200 μM had no effects. Interestingly, NH2Cl did not inhibit IκB kinase activation by TNFα. Protein phosphatase activity did not show apparent change. When recombinant IκB was oxidized by NH2Cl in vitro and phosphorylated by TNFα-stimulated Jurkat cell lysate, its phosphorylation occurred less effectively than non-oxidized IκB. In addition, when NF-κB–IκB complex was immunoprecipitated from NH2Cl-treated cells and phosphorylated in vitro by recombinant active IκB kinase, native IκB but not oxidized IκB was phosphorylated. Amino acid analysis of the in vitro oxidized IκB showed methionine oxidation to methionine sulfoxide. Although Tau–NHCl alone had little effects on TNFα-induced NF-κB activation, simultaneous presence of Tau–NHCl and ammonium ion significantly inhibited the NF-κB activation, probably through the conversion of Tau–NHCl to NH2Cl. These results indicated that NH2Cl inhibited TNFα-induced NF-κB activation through the oxidation of IκB, and that NH2Cl is physiologically more relevant than Tau–NHCl in modifying NF-κB-mediated cellular responses

The whole blood transcriptional regulation landscape in 465 COVID-19 infected samples from Japan COVID-19 Task Force

「コロナ制圧タスクフォース」COVID-19患者由来の血液細胞における遺伝子発現の網羅的解析 --重症度に応じた遺伝子発現の変化には、ヒトゲノム配列の個人差が影響する--. 京都大学プレスリリース. 2022-08-23.Coronavirus disease 2019 (COVID-19) is a recently-emerged infectious disease that has caused millions of deaths, where comprehensive understanding of disease mechanisms is still unestablished. In particular, studies of gene expression dynamics and regulation landscape in COVID-19 infected individuals are limited. Here, we report on a thorough analysis of whole blood RNA-seq data from 465 genotyped samples from the Japan COVID-19 Task Force, including 359 severe and 106 non-severe COVID-19 cases. We discover 1169 putative causal expression quantitative trait loci (eQTLs) including 34 possible colocalizations with biobank fine-mapping results of hematopoietic traits in a Japanese population, 1549 putative causal splice QTLs (sQTLs; e.g. two independent sQTLs at TOR1AIP1), as well as biologically interpretable trans-eQTL examples (e.g., REST and STING1), all fine-mapped at single variant resolution. We perform differential gene expression analysis to elucidate 198 genes with increased expression in severe COVID-19 cases and enriched for innate immune-related functions. Finally, we evaluate the limited but non-zero effect of COVID-19 phenotype on eQTL discovery, and highlight the presence of COVID-19 severity-interaction eQTLs (ieQTLs; e.g., CLEC4C and MYBL2). Our study provides a comprehensive catalog of whole blood regulatory variants in Japanese, as well as a reference for transcriptional landscapes in response to COVID-19 infection

Benefits of intracorporeal gastrointestinal anastomosis following laparoscopic distal gastrectomy

Abstract Background Laparoscopic gastrectomy has recently been gaining popularity as a treatment for cancer; however, little is known about the benefits of intracorporeal (IC) gastrointestinal anastomosis with pure laparoscopic distal gastrectomy (LDG) compared with extracorporeal (EC) anastomosis with laparoscopy-assisted distal gastrectomy (LADG). Methods Between June 2000 and December 2011, we assessed 449 consecutive patients with early-stage gastric cancer who underwent LDG. The patients were classified into three groups according to the method of reconstruction LADG followed by EC hand-sewn anastomosis (LADG + EC) (n = 73), using any of three anastomosis methods (Billroth-I (B-I), Billroth-II (B-II) or Roux-en-Y (R-Y); LDG followed by IC B-I anastomosis (LDG + B-I) (n = 248); or LDG followed by IC R-Y anastomosis (LDG + R-Y) (n = 128)). The analyzed parameters included patient and tumor characteristics, operation details, and post-operative outcomes. Results The tumor location was significantly more proximal in the LDG + R-Y group than in the LDG + B-I group (P P P  Conclusions Intracorporeal mechanical anastomosis by either the B-I or R-Y method following LDG has several advantages over at the LADG + EC, including small wound size, reduced invasiveness, and safe anastomosis. Although additional randomized control studies are warranted to confirm these findings, we consider that pure LDG is a useful technique for patients with early gastric cancer.</p