Impaired plasma clottability induction through fibrinogen degradation by ASP, a serine protease released from Aeromonas sobria

Aeromonas sobria infection often advances to sepsis, in which interaction of bacterial components with plasma proteins possibly causes various disorders. This bacterium releases a serine protease (ASP), a putative virulence factor, and binds to fibrinogen. To study the ASP effect on fibrinogen, we incubated fibrinogen or plasma with ASP and investigated their clotting elicited by thrombin, which converts fibrinogen to a fibrin clot. Enzymatically active ASP retarded plasma clotting in a dose-dependent manner starting at an ASP concentration of 10 nM. ASP also retarded fibrinogen clotting at 3 nM and above, which appeared to correspond to ASP cleavage of fibrinogen at the Aα-chain. Consistent with containing serine protease activity for an ASP-specific substrate, the culture supernatant of an ASP gene-introduced strain retarded plasma and fibrinogen clotting more than that of the wild-type strain. The culture supernatant of an ASP gene-disrupted strain that releases negligible serine protease activity for the ASP-specific substrate did not affect plasma clotting. These results indicate that ASP is the main fibrinogenolytic protease released from A. sobria. Impaired plasma clottability induction through fibrinogen degradation is a new virulence activity of ASP and may contribute to hemorrhagic tendencies in sepsis caused by infection with this bacterium

Development of Bilayered Bone Marrow-derived Cell-Gelatin Grafts for Augmentation Cystoplasty and Reconstruction of Bladder Tissues in Rats

Background : This study attempted to produce a novel graft composed of bone marrow-derived mesenchymal cell (BMC) layer-gelatin sheets for bladder augmentation cystoplasty. Then, we determined if the grafts could reconstruct bladder tissues. Methods : BMCs harvested from the femurs of green fluorescence protein (GFP)-transfected Sprague-Dawley (SD) rats were adherent and proliferating cells on collagen dishes. The cells were then cultured on temperatureresponsive culture dishes. Following this, the BMCs maintaining cell-cell contacts within the monolayer itself were applied to a gelatin sheet. Two BMC layer-gelatin sheets were overlaid together with the cell sides juxtaposed with one another (bilayered BMC-gelatin graft). Bladder top of SD rats were incised and transplanted with the bilayered BMC-gelatin grafts. Similarly, urinary bladders irradiated with 2 Gy once a week for 5 weeks were also conducted. As control, bilayered acellular-gelatin grafts were used. At 4 weeks after transplantation, the bladders were histologically investigated. Results : At 4 weeks after transplantation into either normal or radiation-injured urinary bladders, incised regions closed. The closed regions of bladder top had reconstructed tissues that were formed with urothelium, and smooth muscle layers. Within the reconstructed tissues, the thickness of the smooth muscle layers in the bilayered BMC-gelatin graft-transplanted bladders were larger compared to controls. The GFP-positive transplanted BMCs were detected. Some of the cells were simultaneously positive for smooth muscle or nerve cell markers. Conclusion : This study showed that the bilayered BMC-gelatin grafts that were experimentally produced could reconstruct bladder tissues. The grafts would be developed as grafts for bladder augmentation cystoplasty.Article信州医学雑誌 71(3) : 167-177, (2023)journal articl

Isolation and characterization of a virus (CvV-BW1) that infects symbiotic algae of Paramecium bursaria in Lake Biwa, Japan

<p>Abstract</p> <p>Background</p> <p>We performed an environmental study of viruses infecting the symbiotic single-celled algae of <it>Paramecium bursaria </it>(<it>Paramecium bursaria Chlorella </it>virus, PBCV) in Lake Biwa, the largest lake in Japan. The viruses detected were all <it>Chlorella variabilis </it>virus (CvV = NC64A virus). One of them, designated CvV-BW1, was subjected to further characterization.</p> <p>Results</p> <p>CvV-BW1 formed small plaques and had a linear DNA genome of 370 kb, as judged by pulsed-field gel electrophoresis. Restriction analysis indicated that CvV-BW1 DNA belongs to group H, one of the most resistant groups among CvV DNAs. Based on a phylogenetic tree constructed using the <it>dnapol </it>gene, CvV was classified into two clades, A and B. CvV-BW1 belonged to clade B, in contrast to all previously identified virus strains of group H that belonged to clade A.</p> <p>Conclusions</p> <p>We conclude that CvV-BW1 composes a distinct species within <it>C. variabilis </it>virus.</p

Inhibition of influenza virus replication in cultured cells by RNA-cleaving DNA enzyme

AbstractInfluenza virus replication has been effectively inhibited by antisense phosphothioate oligonucleotides targeting the AUG initiation codon of PB2 mRNA. We designed RNA-cleaving DNA enzymes from 10-23 catalytic motif to target PB2-AUG initiation codon and measured their RNA-cleaving activity in vitro. Although the RNA-cleaving activity was not optimal under physiological conditions, DNA enzymes inhibited viral replication in cultured cells more effectively than antisense phosphothioate oligonucleotides. Our data indicated that DNA enzymes could be useful for the control of viral infection

Interfamily Transfer of Dual NB-LRR Genes Confers Resistance to Multiple Pathogens

A major class of disease resistance (R) genes which encode nucleotide binding and leucine rich repeat (NB-LRR) proteins have been used in traditional breeding programs for crop protection. However, it has been difficult to functionally transfer NB-LRR-type R genes in taxonomically distinct families. Here we demonstrate that a pair of Arabidopsis (Brassicaceae) NB-LRR-type R genes, RPS4 and RRS1, properly function in two other Brassicaceae, Brassica rapa and Brassica napus, but also in two Solanaceae, Nicotiana benthamiana and tomato (Solanum lycopersicum). The solanaceous plants transformed with RPS4/RRS1 confer bacterial effector-specific immunity responses. Furthermore, RPS4 and RRS1, which confer resistance to a fungal pathogen Colletotrichum higginsianum in Brassicaceae, also protect against Colletotrichum orbiculare in cucumber (Cucurbitaceae). Importantly, RPS4/RRS1 transgenic plants show no autoimmune phenotypes, indicating that the NB-LRR proteins are tightly regulated. The successful transfer of two R genes at the family level implies that the downstream components of R genes are highly conserved. The functional interfamily transfer of R genes can be a powerful strategy for providing resistance to a broad range of pathogens

Nagasaki Schizophrenia Study : Outcome of a 15-year Follow-up of an Incident Cohort

The Nagasaki World Health Organization (WHO) Collaborating Center for Research and Training in Mental Health conducted the WHO Coordinated Multi-Center Study of the Long-term Course and Outcome of Schizophrenia as a part of the International Study on Schizophrenia (ISoS). The study used 107 patients who were initially diagnosed as having ICD-9 schizophrenia for the WHO Collaborative Study on the Determinants of Outcome of Severe Mental Disorders (DOSMeD). Subjects were first collected in 1979-1980 for an incidence study of schizophrenia in Nagasaki. In this 15-yearfollow- up study, 7 subjects died, 43 subjects were lost to follow-up and 57 were successfully traced. Among the 7 death cases, 4 suicides were confirmed and 1 was suspected. During the 15-year period, 25 (44%) of the 57 living subjects displayed continuous psychotic course type schizophrenia. During the last 2 years, 14 (25%) were not psychotic ; 31 (54%) were continuously psychotic. Global Assessment of Functioning Scale for Symptomatology (GAF-S) indicated symptomatological outcomes : 16 (28%) had severe symptoms (GAF-S70). Social outcome was evaluated using the Global Assessment of Functioning Scale for Disability (GAF-D): 28 (49%) showed poor adjustment (GAF-D70). The overall time trend was almost evenly divided in thirds : 20 (35%) were getting better, 18 (32%) were the same and 19 (33%) were worse. The present study showed that the outcome of schizophrenia is not always poor, although some patients display a continuous course and poor outcome

Factors Predicting Difficult Biliary Cannulation during Endoscopic Retrograde Cholangiopancreatography for Common Bile Duct Stones

Background/Aims Difficult biliary cannulation is an important risk factor for post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP). Therefore, this study aimed to identify the factors that predict difficult cannulation for common bile duct stones (CBDS) to reduce the risk for PEP. Methods This multicenter retrospective study included 1,406 consecutive patients with native papillae who underwent ERCP for CBDS. Factors predicting difficult cannulation for CBDS were identified using univariate and multivariate analyses. Results Univariate analysis showed that six factors significantly predicted difficult cannulation: ERCP performed by non-expert endoscopists, low-volume center, absence of acute cholangitis, normal serum bilirubin, intradiverticular papilla, and type of major duodenal papilla. Multivariate analysis identified ERCP performed by non-expert endoscopists (odds ratio [OR], 2.5; p<0.001), low-volume center (OR, 1.6; p<0.001), intradiverticular papilla (OR, 1.3; p=0.007), normal serum bilirubin (OR, 1.3; p=0.038), and absence of acute cholangitis (OR, 1.3; p=0.049) as factors significantly predicting difficult cannulation for CBDS. Conclusions Initial cannulation by an experienced endoscopist, early rescue cannulation, or early takeover by an experienced endoscopist should be considered when performing ERCP for CBDS in the presence of factors predicting difficult cannulation

Comparison with Magnetic Resonance Three-Dimensional Sequence for Lumbar Nerve Root with Intervertebral Foramen

Study DesignProspective study based on magnetic resonance (MR) imaging of the lumbar spinal root of the intervertebral foramen.PurposeThis study was to compare MR three-dimensional (3D) sequences for the evaluation of the lumbar spinal root of the intervertebral foramen.Overview of LiteratureThe diagnosis of spinal disorders by MR imaging is commonly performed using two-dimensional T1- and T2-weighted images, whereas 3D MR images can be used for acquiring further detailed data using thin slices with multi-planar reconstruction.MethodsOn twenty healthy volunteers, we investigated the contrast-to-noise ratio (CNR) of the lumbar spinal root of the intervertebral foramen with a 3D balanced sequence. The sequences used were the fast imaging employing steady state acquisition and the coherent oscillatory state acquisition for the manipulation of image contrast (COSMIC). COSMIC can be used with or without fat suppression (FS). We compared these sequence to determine the optimized visualization sequence for the lumbar spinal root of the intervertebral foramen.ResultsFor the CNR between the nerve root and the peripheral tissue, these were no significant differences between the sequences at the entry of foramen. There was a significant difference and the highest CNR was seen with COSMIC-FS for the intra- and extra-foramen.ConclusionsIn this study, the findings suggest that the COSMIC-FS sequences should be used for the internal or external foramen for spinal root disorders

Risk-adjusted therapy for pediatric non-T cell ALL improves outcomes for standard risk patients: results of JACLS ALL-02

This study was a second multicenter trial on childhood ALL by the Japan Childhood Leukemia Study Group (JACLS) to improve outcomes in non-T ALL. Between April 2002 and March 2008, 1138 children with non-T ALL were enrolled in the JACLS ALL-02 trial. Patients were stratified into three groups using age, white blood cell count, unfavorable genetic abnormalities, and treatment response: standard risk (SR), high risk (HR), and extremely high risk (ER). Prophylactic cranial radiation therapy (PCRT) was abolished except for CNS leukemia. Four-year event-free survival (4yr-EFS) and 4-year overall survival (4yr-OS) rates for all patients were 85.4% ± 1.1% and 91.2% ± 0.9%, respectively. Risk-adjusted therapy resulted in 4yr-EFS rates of 90.4% ± 1.4% for SR, 84.9% ± 1.6% for HR, and 66.5% ± 4.0% for ER. Based on NCI risk classification, 4yr-EFS rates were 88.2% in NCI-SR and 76.4% in NCI-HR patients, respectively. Compared to previous trial ALL-97, 4yr-EFS of NCI-SR patients was significantly improved (88.2% vs 81.2%, log rank p = 0.0004). The 4-year cumulative incidence of isolated (0.9%) and total (1.5%) CNS relapse were significantly lower than those reported previously. In conclusion, improved EFS in NCI-SR patients and abolish of PCRT was achieved in ALL-02