Effect of clofibrate treatment on carnitine acyltransferases in different subcellular fractions of rat liver

The subcellular distribution of carnitine acetyl-, octanoyl-, and palmitoyltransferase in the livers of normal and clofibrate-treated male rats was studied with isopycnic sucrose density gradient fraction.In normal liver 48% of total carnitine acetyltransferase activity was peroxisomal, 36% of the activity located in mitochondria and 16% in a membranous fraction containing microsomes. Carnitine octanoyltransferase and carnitine palmitoyltransferase were confined almost totally (77–81%) to mitochondria in normal liver.Clofibrate treatment increased the total activity of carnitine acetyltransferase over 30 times, whereas the total activities of the other two transferases were increased only 5-fold.From the three different subcellular carnitine acetyltransferases the mitochondrial one was not responsive to clofibrate treatment, i.e. the rise in mitochondrial activity was over 70-fold as contrasted to the 6- and 14-fold rises in peroxisomal and microsomal activities, respectively. After treatment mitochondria contained 79% of total activity.It is concluded that the clofibrate-induced increase of carnitine acetyltransferase activity is not due to the peroxisomal proliferation that occurs during clofibrate treatment. The rise in peroxisomal activity contributed only 8% to the total increase.After clofibrate treatment the greatest part of carnitine octanoyl- and palmitoyltrnasferase activities were located in mitochondria but a considerable amount of both activities was found also in the soluble fraction of liver.     

Phorbol myristate acetate stimulation of lymphocyte guanylate cyclase

Human lymphocyte guanylate cyclase activities are increased in a dose-dependent fashion by incubation of intact cells with phorbol myristate acetate, a tumor promoter and lymphocyte mitogen. Increased activity is detectable after 1 minute, and peak membrane-bound and soluble forms of guanylate cyclase occur after 10- and 30-minute exposure to phorbol myristate acetate, respectively. The soluble form is stimulated much more than the membrane form. Enzyme activities measured in the presence of either Ca²⁺, Mg²⁺, or Mn²⁺ are elevated to similar degrees. Comparisons of phorbol and a series of its diesters revealed a good correlation between the capacities for guanylate cyclase stimulation, lymphocyte mitogenesis, and tumor promotion.     

植被的PE（可能蒸散）指标与植被气候分类（三）

Holdridge的生命地带分类系统由于其指标的计算十分简便与对植被的对应性强而受到国际植被生态学界与环境科学研究者的重视。特别是近年来在环境的评价、生态区划与预测全球变化对生态系统的影响等方面得到较多采用。该系统对中国各植被地带的气候台站资料进行计算分析的结果表明有较好的适应性。但由于该系统发展于中美洲的热带地区,因而在中国的亚热带地区须进行局部的调整。但采用该系统将有利于与世界各地的气候—植被分类系统的统一与对比研究。通过回归计算表明,该系统的可能蒸散率(PER)指标与CHIKUGO模型的辐射干燥度(RDI)显著相关。因而可以采用便于取得资料与易于计算的PER来进行潜在第一性生产力(NPP)的估算。对中国各植被地带的计算结果令人满意,可进一步用于在全球变化条件下,中国各植被地带或生态系统主要类型及其NPP变化的预测。     

Predictive markers of safety and immunogenicity of adjuvanted vaccines

Vaccination represents one of the greatest public health triumphs; in part due to the effect of adjuvants that have been included in vaccine preparations to boost the immune responses through different mechanisms. Although a variety of novel adjuvants have been under development, only a limited number have been approved by regulatory authorities for human vaccines. This report reflects the conclusions of a group of scientists from academia, regulatory agencies and industry who attended a conference on the current state of the art in the adjuvant field. Held at the U.S. Pharmacopeial Convention (USP) in Rockville, Maryland, USA, from 18 to 19 April 2013 and organized by the International Association for Biologicals (IABS), the conference focused particularly on the future development of effective adjuvants and adjuvanted vaccines and on overcoming major hurdles, such as safety and immunogenicity assessment, as well as regulatory scrutiny. More information on the conference output can be found on the IABS website, http://www.iabs.org/.     

A Novel and Convenient Method for the Synthesis of Free 5′-Thiol Modified Oligonucleotides

Abstract

The synthesis of free 5′-thiol-modified oligonucleotides using a 4,4′,4″-trimethoxytrityl (TMTr)-protected linker and standard Poly-Pak^TM purification has been described.     

Catalytic DNA and RNA for Targeting MDR1 mRNA

Abstract

Design, synthesis and properties of catalytic NAs for targeting MDR1 mRNA are reported.     

Activation of microsomal glutathione S-transferase activity by sulfhydryl reagents.

Rat liver microsomes exhibit glutathione S-transferase activity with 1-chloro-2,4-dinitrobenzene as the second substrate. This activity can be stimulated 8-fold by treatment of the microsomes with N-ethylmaleimide and 4-fold with iodoacetamide. The corresponding glutathione S-transferase activity of the supernatant fraction is not affected by such treatment. These findings suggest that rat liver microsomes contain glutathione S-transferase distinct from those found in the cytoplasmic and that the microsomal transferase can be activated by modification of microsomal sulfhydryl group(s).     

Proteasomal dysfunction and ER stress triggers 2′-hydroxy-retrochalcone-induced paraptosis in cancer cells

Chalcones are biologically active class of compounds, known for their anticancer activities. Here we show for the first time that out of the six synthetic derivatives of chalcone tested, 2′-hydroxy-retrochalcone (HRC) was the most effective in inducing extensive cytoplasmic vacuolation mediated death called paraptosis in malignant breast and cervical cancer cells. The cell death by HRC is found to be nonapoptotic in nature due to the absence of DNA fragmentation, PARP cleavage, and phosphatidylserine externalization. It was also found to be nonautophagic as there was an increase in the levels of autophagic markers LC3I, LC3II and p62. Immunofluorescence with the endoplasmic reticulum (ER) marker protein calreticulin showed that the cytoplasmic vacuoles formed were derived from the ER. This ER dilation was due to ER stress as evidenced from the increase in polyubiquitinated proteins, Bip and CHOP. Docking studies revealed that HRC could bind to the Thr1 residue on the active site of the chymotrypsin-like subunit of the proteasome. The inhibition of proteasomal activity was further confirmed by the fluorescence based assay of the chymotrypsin-like subunit of the 26S proteasome. The cell death by HRC was also triggered by the collapse of mitochondrial membrane potential and depletion of ATP. Pretreatment with thiol antioxidants and cycloheximide were able to inhibit this programmed cell death. Thus our data suggest that HRC can effectively kill cancer cells via paraptosis, an alternative death pathway and can be a potential lead molecule for anticancer therapy.