Levels of N-acylethanolamines in O,O,S-trimethylphosphorothioate (OOS-TMP)-treated C57BL/6J mice and potential anti-obesity, anti-diabetic effects of OOS-TMP in hyperphagia and hyperglycemia mouse models

O,O,S-Trimethylphosphorothioate (OOS-TMP) has been shown to induce hypophagia and hypopraxia. Recent studies suggest that OOS-TMP-induced anorexia is partly mediated by its effect on the central nervous system. In this study, we examined the profiles of N-acylethanolamines (NEAs), including five amide-linked compounds, in the gastrointestinal system in C57BL/6J (B6) mice. The present results shown an orexigenic profile of the levels of NEAs with downregulation of the anorectic lipid, N-stearoylethanolamine (SEA), upregulation of the orexigenic lipid, 2-arachidonoyl glycerol (2-AG), at 2 h and upregulation of 2-AG at 24 h albeit with significant anorexia. However, the data indicated that the high level of 2-AG may be responsible for the hypopraxia. We next explored whether OOS-TMP may affect two models of hyperphagia and hyperglycemia, ins2^+/Akita B6 (Akita) and B6-lepr^db/lepr^db mice (db/db). We identified potential anorexigenic effects in B6, Akita and db/db mice. Moreover, OOS-TMP was found to reduce blood glucose in Akita mice but not in db/db mice. Collectively, these findings suggest that N-acylethanolamines are not involved in the hypophagia but rather hypopraxia, and may play multiple physiological roles in this process. OOS-TMP might be a promising candidate for anti-obesity and anti-diabetic drug development.     

肺炎支原体抗体IgM检测的常用方法比较

目的比较几种肺炎支原体IgM（MP-IgM）检测方法的敏感性、特异性和准确度，供不同条件的实验室根据实际组合应用。方法酶联免疫吸附试验，金标斑点法，冷凝集试验，间接血凝试验，明胶颗粒凝集法。结果冷凝集试验与其他四种方法存在显著性差异。结论酶联免疫吸附试验敏感性最好，金标斑点法特异性最好，间接血凝法和明胶颗粒凝集法较实用，冷凝集试验诊断意义不大。     

人白细胞介素2-Linker-PE38重组毒素的构建

目的　构建人白介素 2 linker PE38融合基因 ,为进一步表达具有特异性杀伤作用的融合蛋白奠定基础。方法　用PCR方法扩增绿脓杆菌外毒素 (PEA)衍生物PE38基因及柔性连接肽linker片段 ,并与人白介素 2 (Interleukin2 ,IL2 )基因连接插入质粒pET2 8a中。结果　构建的表达载体经核苷酸测序分析无突变发生。结论　成功地构建了表达质粒pIL2 linker Pe38。     

N-Acylethanolamine-Hydrolyzing Acid Amidase Inhibition,but Not Fatty Acid Amide Hydrolase Inhibition,Prevents the Development of Experimental Autoimmune Encephalomyelitis in Mice

N-acylethanolamines (NAEs) are endogenous bioactive lipids reported to exert anti-inflammatory and neuroprotective effects mediated by cannabinoid receptors and peroxisome proliferator–activated receptors (PPARs), among others. Therefore, interfering with NAE signaling could be a promising strategy to decrease inflammation in neurological disorders such as multiple sclerosis (MS). Fatty acid amide hydrolase (FAAH) and N-acylethanolamine-hydrolyzing acid amidase (NAAA) are key modulators of NAE levels. This study aims to investigate and compare the effect of NAAA inhibition, FAAH inhibition, and dual inhibition of both enzymes in a mouse model of MS, namely the experimental autoimmune encephalomyelitis (EAE). Our data show that NAAA inhibition strongly decreased the hallmarks of the pathology. Interestingly, FAAH inhibition was less efficient in decreasing inflammatory hallmarks despite the increased NAE levels. Moreover, the inhibition of both NAAA and FAAH, using a dual-inhibitor or the co-administration of NAAA and FAAH inhibitors, did not show an added value compared to NAAA inhibition. Furthermore, our data suggest an important role of decreased activation of astrocytes and microglia in the effects of NAAA inhibition on EAE, while NAAA inhibition did not affect T cell recall. This work highlights the beneficial effects of NAAA inhibition in the context of central nervous system inflammation and suggests that the simultaneous inhibition of NAAA and FAAH has no additional beneficial effect in EAE.Supplementary InformationThe online version contains supplementary material available at 10.1007/s13311-021-01074-x.     

Monoamine oxidase activities of porcine vascular endothelial and smooth muscle cells

Amine uptake by cultured vascular cells was studied under conditions minimizing nonenzymic oxidation. 5-Hydroxytryptamine (5HT) was accumulated only very poorly; detailed kinetic analysis couid not be performed, but there was no evidence for a saturable high affinity process. Comparison of β-phenylethylamine (PEA) and 5HT metabolism in intact cells and lysed cells demonstrated that the rates of entry of the amines into cells usually limited their metabolism especially at low (μM) concentrations. Primary cultures of aortic endothelial cells metabolised 5HT and PEA substantially faster than did subcultured endothelium. Subcultured aortic vascular smooth muscle cells and endothelial cells metabolised PEA and 5HT with comparable specific enzyme activities to those found in aortic medial tissue. Inhibition by clorgyline of PEA, 5HT and benzylamine (BZA) metabolism reveaied, however, that while aortic tissue possessed monoamine oxidase (MAO) types A and B and a comparable amount of a clorgyline resistant amine oxidase(s) (CRAO), cultured vascular cells possessed MAO-A, but little or no CRAO or MAO-B. Cultured venous endothelium, and smooth muscle from several vascular sites, metabolised PEA and 5HT at similar rates to those found in aortic cells. the studies demonstrate that although cultured porcine endothelial and smooth muscle cells from large blood vessels contain MAO, they do not apparently possess the amine transport process present in the lung. Additionally, conditions of culture can affect both the extent of amine metabolism and the pattern of amine oxidase present.     

Role of Ets transcription factors in mammary gland development and oncogenesis

PEA3 is the founding member of a subfamily of closely related ets genes that includes ER81 and ERM. PEA3 is expressed in the epithelial cells of mammary buds at the time that these first appear during mouse embryogenesis, and it is differentially expressed during postnatal mammary gland development. PEA3 expression is highest at the onset of puberty and during early pregnancy, times of extensive epithelial outgrowth and branching. PEA3 is expressed in undifferentiated epithelial cap cells of terminal end buds, and in differentiated myoepithelial cells of ducts and alveoli. Loss-of-function mutations in the PEA3 gene compromise mammary ductal branching at the onset of puberty and early during pregnancy. PEA3 is overexpressed in the vast majority of human breast tumors and in nearly all of the HER2-positive subclass of such tumors. PEA3 is similarly overexpressed in transgenic mouse models of this malignancy. Expression of dominant-negative PEA3 in the mouse mammary gland of MMTV-HER2 transgenic mice dramatically delays the onset and reduces the incidence of mammary tumors. Hence PEA3 and/or its close relatives play key regulatory roles in both mammary gland development and oncogenesis.