Effect of glucose on protein phosphorylation in rat pancreatic islets

Isolated rat pancreatic islets, incubated in the presence of extracellular ³²Pi to steady state ³²P incorporation into cellular phosphopeptides, were exposed to glucose for 10 min. Glucose (16.7 mM) significantly stimulated the phosphorylation of six phosphoproteins with molecular weights of 15,000, 35,000, 49,000, 64,000, 93,000 and 138,000. Mannoheptulose (16.7 mM) markedly inhibited glucose-stimulated phosphorylation of these six phosphoproteins. This protein phosphorylation might be important in mediating glucose-stimulated insulin release.     

Hemocytes of Bulinus truncatus rohlfsi (Mollusca: Gastropoda)

Two basic cell types occur in the hemolymph of Bulinus truncatus rohlfsi: granulocytes and hyalinocytes. Granulocytes are divided into three subtypes: (1) Granulocytes I, which account for 19% of the hemocytes, are small, young amoebocytes with 1–20 filopodia and small numbers of cytoplasmic granules, including some lysosomes; (2) granulocytes II, which account for 78% of the cells, are large, fully developed amoebocytes that possess 1–20 filopodia and many granules, both acidophilic and basophilic, including numerous lysosomes, phagosomes, and mitochondria; and (3) spent granulocytes, which are rare, have few filopodia, large accumulations of glycogen granules and prominent vacuoles in addition to lysosomes in the cytoplasm. These three subtypes of granulocytes probably represent ontogenetic stages within a single cell line. In addition, granulocytes with 40 or more filopodia and little ectoplasm, found in only 1 of 45 snails examined, probably reflect a pathologic condition. Hyalinocytes, which account for 3% of all hemocytes, are similar in size to mature granulocytes, but have few or no cytoplasmic granules and lack filopodia and glycogen granules. Total hemocyte concentration in hemolymph is 328,000 ± 188,000 cells/ml.     

HISTOLOGICAL ANALYSIS OF ADVENTITIOUS BUD FORMATION IN CULTURED DOUGLAS FIR COTYLEDON

Initiation of adventitious bud formation in vitro from Douglas fir cotyledons required both cytokinin and auxin at concentrations of 5 μM BAP and 5 nM NAA. Histological observations showed that these adventitious buds arose de novo from cells residing in hypodermal layers. Development of adventitious buds in culture was characterized by the sequential appearance of four anatomically distinguishable structures: 1) meristemoid, 2) bud primordium, 3) shoot apex with needle primordia, and 4) adventitious bud. The anatomical structure of tissue culture-produced buds was similar to that of vegetative buds produced on intact plants. Cultured cotyledons capable of producing adventitious buds (bud culture) were compared with bud-callus and callus cultures initiated by 5 μM BAP plus 5 μM NAA and 5μM NAA alone without BAP, respectively. Results showed that, during early stages of the culture period (i.e., prior to the appearance of meristemoid structure), cell division of bud culture was mainly located in hypodermal layers, whereas for the other culture types, bud-callus and callus cultures, cell division occurred randomly in all tissues.     

Spectrophotometric determination of bromopyruvate by reaction with 2-nitro-5-thiobenzoic acid

A spectrophotometric method for determination of bromopyruvate, based on the reaction with 2-nitro-5-thiobenzoic acid, is described. The reaction is complete in 30 min at room temperature in 0.1 m Tris-MES, 1 mm EDTA, pH 8.0. The method is sensitive to at least 1 × 10^?5m bromopyruvate. Reagents are stable, easy to prepare, and specific for β-halopyruvate. Bromopyruvate solutions must be prepared fresh daily. Solutions of bromopyruvate at pH 8.0 and 23°C have a half-life of 3 hr.     

The inhibition of sugar transport and oxidation in fat cell ghosts by colchicine.

Colchicine inhibits glucose oxidation and the uptake of 2-deoxy-D-glucose in fat cell ghosts but has no effect on glucose oxidation by fat cell homogenates. This inhibition is rapid, reversible, and temperature-independent. Insulin-stimulated glucose oxidation and 2-deoxy-D-glucose transport are also inhibited by colchicine to an extent comparable to the basal processes.     

SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target

Cytokine storm and multi-organ failure are the main causes of SARS-CoV-2-related death. However, the origin of excessive damages caused by SARS-CoV-2 remains largely unknown. Here we show that the SARS-CoV-2 envelope (2-E) protein alone is able to cause acute respiratory distress syndrome (ARDS)-like damages in vitro and in vivo. 2-E proteins were found to form a type of pH-sensitive cation channels in bilayer lipid membranes. As observed in SARS-CoV-2-infected cells, heterologous expression of 2-E channels induced rapid cell death in various susceptible cell types and robust secretion of cytokines and chemokines in macrophages. Intravenous administration of purified 2-E protein into mice caused ARDS-like pathological damages in lung and spleen. A dominant negative mutation lowering 2-E channel activity attenuated cell death and SARS-CoV-2 production. Newly identified channel inhibitors exhibited potent anti-SARS-CoV-2 activity and excellent cell protective activity in vitro and these activities were positively correlated with inhibition of 2-E channel. Importantly, prophylactic and therapeutic administration of the channel inhibitor effectively reduced both the viral load and secretion of inflammation cytokines in lungs of SARS-CoV-2-infected transgenic mice expressing human angiotensin-converting enzyme 2 (hACE-2). Our study supports that 2-E is a promising drug target against SARS-CoV-2.Subject terms: Cell death, Molecular biology     

金合欢根瘤菌的几种酶活特性

金合欢根瘤菌Rhizobium sp.(Acacia farnesiana)的氧化酶、过氧化氢酶、脲酶、青霉素酶和β-半乳糖苷酶均呈阳性。经过酶活性定量测定表明金合欢根瘤菌有葡萄糖酸-6-磷酸脱氢酶(6PGD)和β-半乳糖苷酶的酶活性,而慢生型大豆根瘤菌未测到上述两种酶活性。根据聚丙烯酰胺凝胶电泳酯酶图谱,6株金合欢根瘤菌可分成两群:酋株AF1、AF2和AF3的酯酶图谱中有A带,AF4、AF5和AF6的酯酶图谱中没有A带。