エラスターゼ産生性Prevotella intermediaの菌体外ゼラチナーゼの分離と性状

Gelatinase in the culture supernatant of Prevotella intermedia was purified to homogeneity by the combined procedure of ethanol precipitation and three steps of chromatography. The enzyme was a cystein protease with a molecular mass of 45 kDa. The activity was inhibited by divalent metal chelators and its inhibition was recovered by the addition of Ca^. The optimum pH for activity was 7.0-7.5 and the enzyme was inactivated by heating at 60℃ for 10 min. It hydrolyzed actively azocoll and hide powder besides gelatin. Hydrolysis of type IV collagen, if not strong, was also observed

口腔スピロヘータTreponema denticolaの分離とその生物的性状

Spirochetes were isolated from the lesions of periodontitis patients, identified them as Treponema denticola and their properties were investigated. These strains required serum for growth and demonstrated low sugar fermentation activity. Indole, hydrogen sulfide, acetic acid, propionic acid and butyric acid were detected in the metabolites of all the isolates. Proteases, esterase and neuraminidase were also produced by the isolates, of which, proteolytic enzymes appeared to contain at least three different kinds. These proteases were possible to separate from each other by column chromatography

Acid Dissociation Equilibrium and Singlet Molecular Oxygen Quantum Yield of Acetylated 6,8–Dithioguanosine in Aqueous Buffer Solution

2’,3’,5’–Tri–O–acetyl–6,8–dithioguanosine (taDTGuo) is a modified nucleoside of drug 6–thioguanine and further developed as a potential photochemotherapeutic agent due to its desirable properties of photosensitivity to UVA light and singlet molecular oxygen generation. The photochemical characteristics of taDTGuo under biological conditions (namely in aqueous solution) were intensively investigated by the steady–state absorption and emission, time–resolved near–infrared emission measurements, and quantum chemical calculations. taDTGuo was found to be held in sequential acid dissociation equilibria within pH 3.79–11.93. With the global fitting analysis of the absorption spectra at various pHs, two pKa values of the equilibria were determined to be 7.02 ± 0.01 and 9.79 ± 0.01. Quantum chemical calculations suggested that its mono– and di–anionic species in the ground state should be 1–imide anionic form (N1–taDTGuo–) and 1,7–di–imide anionic form (taDTGuo2–) respectively. taDTGuo generates singlet molecular oxygen effectively and has pH–dependent quantum yields. In conclusion, taDTGuo would be most useful as a potent agent for photochemotherapy under certain carcinomatous pH conditions

Physiological and Pathological Mitochondrial Clearance Is Related to Pectoralis Major Muscle Pathogenesis in Broilers With Wooden Breast Syndrome

Wooden breast syndrome (WB) constitutes an emerging myopathy in the pectoralis major muscle (PM) of broiler chickens, characterized by myofiber hypertrophy and degeneration along with severe fibrosis. WB pathogenesis has been considered to involve hypoxia induced by rapid growth of the PM. In this study, we focused on mitochondrial morphology and dynamics in the myofibers, as these organelles are sensitive to damage by hypoxia, and examined the effects on WB pathogenesis. Specifically, the PMs of a flock of 35 broilers at 50 days of age were evaluated. First, the severity of disease in each bird was determined by measuring histopathological indices including the fibrotic area (FA) in the muscle and circularity of myofibers (CM). These values were 29.4 ± 9.6% and 0.70 ± 0.042, respectively, showing variety among the flock. Myofiber vacuolization was observed in all birds including numerous small- or large-rimmed vacuoles, with the former consisting of ultrastructurally autophagosome-like vacuoles engulfing degenerated mitochondria. The large-rimmed vacuoles frequently occurred in the PMs with more severe FA and CM, indicating a relationship between altered autophagy/mitophagy and WB severity. Next, the expression levels of hypoxia-adaptive and mitochondrial dynamics-related genes were analyzed, and their correlations with the histopathological indices were examined. The histopathological indices were negatively correlated with the expression of vascular endothelial growth factor A (VEGFA), indicating that less angiogenesis owing to weakened hypoxia-inducible factor signaling induces more severe WB pathology. In addition, the observed negative correlation with mitochondrial dynamics-related genes implied that WB pathology deteriorates concomitant with reduced mitochondrial dynamics. Furthermore, the expression of mitochondrial dynamics-related genes showed strong positive correlation with that of VEGFA and autophagy-/mitophagy-related genes. These results revealed that the PMs of broilers possess the mechanism of physiological clearance of mitochondria damaged by the hypoxia resulting from the continuous mitochondrial dynamics and autophagy/mitophagy accompanying rapid PM growth. In turn, the altered mitochondrial clearance induced by chronic hypoxia and the accumulation of damaged mitochondria likely underly the severe pathological features of WB.ArticleFrontiers in Pharmacology.11:579(2020)journal articl

Early treatment with a sodium-glucose co-transporter 2 inhibitor in high-risk patients with acute heart failure:Rationale for and design of the EMPA-AHF trial

Aims: The aim of the EMPA-AHF trial is to clarify whether early initiation of a sodium-glucose co-transporter 2 inhibitor before clinical stabilization is safe and beneficial for patients with acute heart failure (AHF) who are at a high risk of adverse events. Methods: The EMPA-AHF trial is a randomized, double-blind, placebo-controlled, multicentre trial examining the efficacy and safety of early initiation of empagliflozin (10 mg once daily). In total, 500 patients admitted for AHF will be randomized 1:1 to either empagliflozin 10 mg daily or placebo at 47 sites in Japan. Study entry requires hospitalization for AHF with dyspnoea, signs of volume overload, elevated natriuretic peptide, and at least one of the following criteria: estimated glomerular filtration rate &lt;60 mL/min/1.73 m2; already taking ≥40 mg of furosemide daily before hospitalization; and urine output of &lt;300 mL within 2 hours after an adequate dose of intravenous furosemide. Patients will be randomized within 12 hours of hospital presentation, with treatment continued up to 90 days. The primary outcome is the clinical benefit of empagliflozin on the win ratio for a hierarchical composite endpoint consisting of death within 90 days, heart failure rehospitalization within 90 days, worsening heart failure during hospitalization, and urine output within 48 hours after treatment initiation. Conclusion: The EMPA-AHF trial is the first to evaluate the efficacy and safety of early initiation of empagliflozin in patients with AHF considered to be at high risk under conventional treatment.</p