Organelle contacts: Sub‐organelle zones to facilitate rapid and accurate inter‐organelle trafficking of lipids

When one person wants to communicate securely with another, he/she should contact the other person directly. This rule applies not only to human society, but also to the intracellular micro‐society. In the past two decades, it has become increasingly clear that the sub‐organelle regions called membrane contact sites (MCSs) are pivotal for inter‐organelle transport of lipids in cells, as highlighted in the thematic review series “Interorganelle trafficking of lipids” held in Traffic in 2014–2015. In this commentary, we will describe how the currently prevailing model for lipid trafficking at MCSs was generated, and comment on three important issues that have not been explored: (a1) the principles guiding the generation of an asymmetrical inter‐organelle flow of lipids in cells, (b2) the advantages in lipid trafficking at organelle contacts, and (c3) the dynamic network of inter‐organelle lipid trafficking.     

Erythroleukemia treated effects of rat plasma profile and erythrocyte membranes

Erythroleukemia disease is caused by over production of malignant blood and immature large number of blood cells enters into peripheral compartment. Biophysical and biochemical changes in plasma and erythrocyte membrane in erythroleukemia treated rats were identified. Our study, leukemia is experimentally exposed in rats were injecting erythroleukemia cells (FLC) (H-2d) intravenously in adult rats and normal control rats were maintained. Significant increase in the activity of blood glucose, proteins levels, aspartate transaminase (AST) and alanine transaminase (ALT) values and significant decrease in haemoglobin (Hb), albumin levels in erythroleukemia treated rats were observed when compared with control rats. Cholesterol and low density liproprotein (LDL) levels increased significantly in erythroleukemia treated rats but triglycerides, high density lipoprotein (HDL) and very low density lipoprotein (VLDL) levels decreased significantly. Levels of red cell membrane cholesterol decreased in erythroleukemia treated rats in comparison with control while levels of phospholipids and proteins increased in erythrocytes of erythroleukemia treated rats. Red blood cell (RBC) and white blood cell (WBC) counts increased significantly and platelet count decreased. C/P (cholesterol/phospholipid) ratio decreased significantly in erythroleukemia treated rats. This study has been undertaken for the first time to investigate the effect of (FLC) (H-2d) erythroleukemia cells (treated) in intravenously in adult rats and normal control rats. Results indicate biophysical and biochemical alterations at molecular level in plasma and erythrocyte membrane.     

他汀类药物对中老年男性冠心病患者肠道菌群的影响

目的探讨他汀类药物对中老年男性稳定型冠心病患者肠道菌群的影响,为临床冠心病的二级预防提供参考。方法选择64例经皮冠状动脉介入治疗术后坚持服用中等强度他汀类药物的男性稳定型冠心病患者为研究对象。患者年龄55～74岁,均不合并糖尿病。根据研究期间患者低密度脂蛋白胆固醇(LDL-C)水平分为血脂未达标组(NSG组,LDL-C≥1.8 mmol/L)与血脂达标组(SG组,LDL-C1.8 mmol/L)。比较两组患者一般临床情况,治疗前后血糖、血脂水平,同时应用16S rRNA测序分析技术对两组患者肠道菌群进行分析。结果两组患者一般临床情况及他汀类药物治疗前患者血糖、血脂水平差异均无统计学意义(均P0.05)。治疗后,NSG组患者总胆固醇、LDL-C、三酰甘油水平均明显高于SG组;同时NSG组患者治疗后空腹血糖水平明显高于治疗前(均P0.05)。Alpha多样性分析显示,NSG组患者肠道菌群多样性低于SG组(P0.05)。LEfSe分析显示,SG组患者肠道Verrucomicrobia、Verrucomicrobiae、Verrucomicrobiales、Verrucomicrobiaceae、Akkermansias、Akkermansia muciniphila、Subdoligranulum和uncultured_Christensenellaceae_bacterium数量明显高于NSG组,而uncultured_Veillonellaceae_bacterium及Prevotella_stercorea的丰度显著低于NSG组。结论中等强度他汀类药物对男性中老年稳定型冠心病患者的疗效与肠道菌群存在一定关系,主要表现为治疗后LDL-C未达标者肠道脂代谢有益菌Akkermansia muciniphila及Subdoligranulum丰度明显减少。     

Arylesterase activity of paraoxonase 1 (PON1) on HDL3 and HDL2: Relationship with Q192R,C-108T,and L55M polymorphisms

BackgroundControversy exists regarding the role of the subfractions of high-density lipoproteins (HDL₂ and HDL₃) in cardiovascular disease. The functionality of these particles, and their protective role, is due in part to the paraoxonase 1 (PON1) presence in them. The polymorphisms rs662 (Q192R, A/G), rs854560 (L55 M, T/A), and rs705379 (C-108T) of the PON1 gene have been related to enzyme activity and, with the anti-oxidative capacity of the HDL. The objective was to determine the arylesterase PON1 activity in HDL₃ and HDL₂ and its relationship with the polymorphisms mentioned, in a young population.MethodsThe polymorphisms were determined through mini-sequencing (SnaPshot). The HDL subpopulations were separated via ionic precipitation, cholesterol was measured with enzymatic methods, and PON1 activity was measured through spectrophotometry.ResultsThe results show that the PON1 polymorphisms do not influence the cholesterol in the HDL. A variation between 40.02 and 43.9 mg/dL was in all the polymorphisms without significant differences. Additionally, PON1 activity in the HDL₃ subfractions was greater (62.83 ± 20 kU/L) than with HDL₂ (35.8 ± 20.8 kU/L) in the whole population and in all the polymorphisms (p < 0.001), and it was independent of the polymorphism and differential arylesterase activity in the Q192R polymorphism (QQ > QR > RR). Thus, 115.90 ± 30.7, 88.78 ± 21.3, 65.29 ± 10.2, respectively, for total HDL, with identical behavior for HDL₃ and HDL₂.ConclusionsPON1 polymorphisms do not influence the HDL_-c, and the PON activity is greater in the HDL₃ than in the HDL₂, independent of the polymorphism, but it is necessary to delve into the functionality of these findings in different populations.     

Performance of Luffa cylindrica as an immobilization matrix for the biotransformation of cholesterol by Mycobacterium species

Abstract

The microbial cleavage of the side chain of cholesterol is a slow process due to the low solubility of the substrate in aqueous media (< 1 μM). Cell immobilization has been shown to be an efficient technology for enhancing the yield of cholesterol biotransformation. In these experiments, living cells of Mycobacterium sp. DSM 2966 and Mycobacterium sp. DSM 2967 were immobilized by passive adsorption on different types of solid carriers. As compared to the control and other solid supports, Luffa cylindrica resulted in a 3–4-fold increase of the specific side chain cleavage activity after 7 days of incubation. Luffa cylindrica had no significant negative influence on cell growth. Furthermore, it is a natural, inexpensive, non-toxic and mechanically strong material and therefore suitable for follow-up experiments.     

Columbin inhibits cholesterol uptake in bloodstream forms of Trypanosoma brucei-A possible trypanocidal mechanism

The diterpenoid furanolactone (columbin) from Aristolochia albida inhibited growth of culture forms of Trypanosoma brucei. In vitro analysis of the compound at 5–250 μg/ml showed complete lysis of the parasites within 10–20 minutes post incubation. At 50 μg/ml, columbin killed about 50% of the parasites which initially appeared swollen under phase contrast microscopy. Also the total amount of cholesterol diminished dose-dependently in the presence of 10–100 μg/ml of columbin after a 3-day incubation period.

In vivo analysis of the compound in T. brucei-infected mice revealed that 25 mg/kg administered for 3 consecutive days, completely cleared the parasites from the peripheral circulation. However, columbin could not clear parasites in the cerebrospinal fluid.     

The AAA ATPase VPS4/SKD1 Regulates Endosomal Cholesterol Trafficking Independently of ESCRT‐III

The exit of low‐density lipoprotein derived cholesterol (LDL‐C) from late endosomes (LE)/lysosomes (Ly) is mediated by Niemann–Pick C1 (NPC1), a multipass integral membrane protein on the limiting membranes of LE/Ly, and by NPC2, a cholesterol‐binding protein in the lumen of LE/Ly. NPC2 delivers cholesterol to the N‐terminal domain of NPC1, which is believed to insert cholesterol into the limiting membrane for subsequent transport to other subcellular organelles. Few cytoplasmic factors have been identified to govern cholesterol efflux from LE/Ly, and much less is known about the underlying molecular mechanisms. Here we establish VPS4, an AAA ATPase that has a well‐established role in disassembling the ESCRT (endosomal sorting complex required for transport)‐III polymer, as an important regulator of endosomal cholesterol transport. Knocking down VPS4 in HeLa cells resulted in prominent accumulation of LDL‐C in LE/Ly, and disrupted cholesterol homeostatic responses at the endoplasmic reticulum. The level and localization of NPC1 and NPC2 appeared to be normal in VPS4 knockdown cells. Importantly, depleting any of the ESCRT‐III components did not exert a significant effect on endosomal cholesterol transport. Our results thus identify an important cytoplasmic regulator of endosomal cholesterol trafficking and represent the first functional separation of VPS4 from ESCRT‐III.     

Lysosomal Membrane Proteins and Their Central Role in Physiology

The lysosomal membrane was thought for a long time to primarily act as a physical barrier separating the luminal acidic milieu from the cytoplasmic environment. Meanwhile, it has been realized that unique lysosomal membranes play essential roles in a number of cellular events ranging from phagocytosis, autophagy, cell death, virus infection to membrane repair. This review provides an overview about the most interesting emerging functions of lysosomal membrane proteins and how they contribute to health and disease. Their importance is exemplified by their role in acidification, transport of metabolites and ions across the membrane, intracellular transport of hydrolases and the regulation of membrane fusion events. Studies in patient cells, non‐mammalian model organisms and knockout mice contributed to our understanding of how the different lysosomal membrane proteins affect cellular homeostasis, developmental processes as well as tissue functions. Because these proteins are central for the biogenesis of this compartment they are also considered as attractive targets to modulate the lysosomal machinery in cases where impaired lysosomal degradation leads to cellular pathologies. We are only beginning to understand the complex composition and function of these proteins which are tightly linked to processes occurring throughout the endocytic and biosynthetic pathways.