Effect of epinephrine on glycogen phosphorylase-alpha in various preparations of rat liver

1. Glycogen phosphorylase-alpha, a commonly used index of cytosolic free calcium, was compared in various preparations of rat liver in the absence and presence of 0.1 microM epinephrine. 2. Total phosphorylase in isolated perfused livers and freshly-isolated hepatocytes were the same as that observed in liver in situ; however, phosphorylase-alpha was 50% higher in perfused liver and 80% higher in hepatocytes than activities measured in situ. Total phosphorylase was reduced approximately 50% in hepatocytes maintained in primary culture for 24 hr. 3. Epinephrine increased phosphorylase-alpha approximately 2-fold in livers perfused for 30 min but only about 20% in hepatocytes incubated for 30 min. After 90 min of perfusion or incubation, epinephrine increased phosphorylase-alpha nearly 4-fold in perfused livers and only 30% in isolated hepatocytes. The results suggest that amounts of free calcium and calcium-dependent coupling of adrenergic receptors to phosphorylase-alpha differ markedly between the intact liver and isolated hepatocytes.     

Quantitative and phenotypic analysis of bone marrow-derived cells in the intact and inflamed central nervous system

Bone marrow has been proposed as a possible source of cells capable of replacing injured neural cells in diseases such as Multiple Sclerosis (MS). Previous studies have reported conflicting results regarding the transformation of bone marrow cells into neural cells in vivo. This study is a detailed analysis of the fate of bone marrow derived cells (BMDC) in the CNS of C57Bl/6 mice with and without experimental autoimmune encephalomyelitis using flow cytometry to identify GFP-labeled BMDC that lacked the pan-hematopoietic marker CD45 and co-expressed neural markers polysialic acid-neural cell adhesion molecule or A2B5. A small number of BMDC displaying neural markers and lacking CD45 expression was identified within both the non-inflamed and inflamed CNS. However, the majority of BMDC exhibited a hematopoietic phenotype.Key words: bone marrow, transplantation, transdifferentiation, central nervous system, green fluorescence protein, experimental autoimmune encephalomyelitis, multiple sclerosis     

中药新药保肝作用的研究进展

肝脏作为代谢器官,在人体内发挥着重要作用。随着肝病的发病率逐年上升,如何有效的保肝护肝已成为医学界和药学界共同面一临的巨大挑战之一。化学药物在治疗肝病的同时常常伴随各种毒副作用甚至更进一步的肝损伤,而中药凭借其安全性和有效性的优势在肝病治疗领域受到越来越多的重视。中药护肝已有悠久的历史,近年来随着技术的进步,更多更好的新型中药逐步上市,相关研究不断增多。本文将就这些研究成果进行阐述。     

Development of potent autophagy inhibitors that sensitize oncogenic BRAF V600E mutant melanoma tumor cells to vemurafenib

Autophagy is a dynamic cell survival mechanism by which a double-membrane vesicle, or autophagosome, sequesters portions of the cytosol for delivery to the lysosome for recycling. This process can be inhibited using the antimalarial agent chloroquine (CQ), which impairs lysosomal function and prevents autophagosome turnover. Despite its activity, CQ is a relatively inadequate inhibitor that requires high concentrations to disrupt autophagy, highlighting the need for improved small molecules. To address this, we screened a panel of antimalarial agents for autophagy inhibition and chemically synthesized a novel series of acridine and tetrahydroacridine derivatives. Structure-activity relationship studies of the acridine ring led to the discovery of VATG-027 as a potent autophagy inhibitor with a high cytotoxicity profile. In contrast, the tetrahydroacridine VATG-032 showed remarkably little cytotoxicity while still maintaining autophagy inhibition activity, suggesting that both compounds act as autophagy inhibitors with differential effects on cell viability. Further, knockdown of autophagy-related genes showed no effect on cell viability, demonstrating that the ability to inhibit autophagy is separate from the compound cytotoxicity profiles. Next, we determined that both inhibitors function through lysosomal deacidification mechanisms and ultimately disrupt autophagosome turnover. To evaluate the genetic context in which these lysosomotropic inhibitors may be effective, they were tested in patient-derived melanoma cell lines driven by oncogenic BRAF (v-raf murine sarcoma viral oncogene homolog B). We discovered that both inhibitors sensitized melanoma cells to the BRAF V600E inhibitor vemurafenib. Overall, these autophagy inhibitors provide a means to effectively block autophagy and have the potential to sensitize mutant BRAF melanomas to first-line therapies.     

The influence of oxygenated sterol compounds on dipalmitoylphosphatidylcholine bilayer structure and packing

Fourier Transform Infra-red and Raman Spectroscopies indicate that 7 alpha-hydroxycholesterol and 7-ketocholesterol have a diminished capacity to condense (increase the packing order of) fluid-state dipalmitoylphosphatidylcholine (DPPC) acyl chains when compared with the effects of cholesterol and the other oxidized sterols studied. DPPC head groups were also more ordered by 7-ketocholesterol over the temperature range 10 degrees - 70 degrees C. Primary effects of these sterols appear to be associated with the hydrophillic regions of the DPPC bilayer, although packing arrangements with acyl chains are also involved. Phosphate and acyl chain ester groups were observed to possess a packing order which was invariant which indicates that these may be the target groups in the interaction with 7-ketocholesterol. A surprising observation was the synergistic amplification of the effects of 7-ketocholesterol by the presence of cholesterol in the DPPC bilayer.     

Blood flow and gastric secretion

The relationship between gastric blood flow and acid secretion has been studied by using a number of secretory stimulants and inhibitors and different techniques that measure gastric blood flow. Although there are conflicting data, there appears to be a consensus regarding the main aspects of this relationship. Agents that stimulate gastric acid secretion such as histamine, gastrin, cholinergic agents, and vagal stimulators also increase gastric blood flow. Other agents such as isoproterenol, epinephrine, and prostaglandins, which at low doses increase gastric blood flow, reduce gastric acid secretion at higher doses. Norepinephrine, vasopressin, and shock reduce gastric blood flow and thereby cause a decrease in secretion. Histamine H2-receptor antagonists reduce stimulated acid secretion and gastric blood flow. Histamine, gastrin, and acetylcholine have been shown to stimulate acid secretion in vitro. Therefore, these observations suggest that although blood flow is not a prerequisite for initiation of stimulated acid secretion, it can become rate-limiting at higher rates of secretion. Although the literature is replete with studies that attempt to characterize the relationship between gastric blood flow and acid secretion, conclusions have varied. Much of the difficulty has arisen because of the differences in technique used to measure gastric blood flow and the differences between anesthetized and unanesthetized animal preparations. Under some specific conditions, the different blood flow techniques give comparable results and this relationship can be defined.     

Shrimp ponds lead to massive loss of soil carbon and greenhouse gas emissions in northeastern Brazilian mangroves

Mangroves of the semiarid Caatinga region of northeastern Brazil are being rapidly converted to shrimp pond aquaculture. To determine ecosystem carbon stocks and potential greenhouse gas emissions from this widespread land use, we measured carbon stocks of eight mangrove forests and three shrimp ponds in the Acaraú and Jaguaribe watersheds in Ceará state, Brazil. The shrimp ponds were paired with adjacent intact mangroves to ascertain carbon losses and potential emissions from land conversion. The mean total ecosystem carbon stock of mangroves in this semiarid tropical landscape was 413 ± 94 Mg C/ha. There were highly significant differences in the ecosystem carbon stocks between the two sampled estuaries suggesting caution when extrapolating carbon stock across different estuaries even in the same landscape. Conversion of mangroves to shrimp ponds resulted in losses of 58%–82% of the ecosystem carbon stocks. The mean potential emissions arising from mangrove conversion to shrimp ponds was 1,390 Mg CO₂e/ha. Carbon losses were largely from soils which accounted for 81% of the total emission. Losses from soils >100 cm in depth accounted for 33% of the total ecosystem carbon loss. Soil carbon losses from shrimp pond conversion are equivalent to about 182 years of soil carbon accumulation. Losses from mangrove conversion are about 10‐fold greater than emissions from conversion of upland tropical dry forest in the Brazilian Caatinga underscoring the potential value for their inclusion in climate change mitigation activities.