(A)GGG(A), (A)CCC(A) and other potential 3′ splice signals in primate nuclear pre-mRNA sequences

Several 3′ splice signals in nuclear precursor mRNAs have already been known for some time: the AG doublet on the left-hand side of the splice and a run of pyrimidines just upstream of it. More recently it has been noted that the YNYTRAY sequence (where Y is a pyrimidine, R a purine and N any base) is a branching-sequence participating in formation of a lariat structure. Keller and Noon have shown the existence of several putative consensus sequences at this site. In this work, extensive computations of the distributions of 256 quartets in all primate nuclear pre-mRNA intron sequences present in GenBank have been carried out. Several putative signals upstream and downstream of the 3′ splice have been detected. These have been compared with the results obtained in analogous computations carried out on all nuclear pre-mRNA introns present in a combined eukaryotic file containing mammal, non-mammalian vertebrate, invertebrate and plant sequences. The distributions of the more interesting oligomers are shown here. Of particular interest are the putative (A)GGG(A) signal 60 nucleotides upstream of the 3′ splice site and (A)CCC(A) 3–40 nucleotides downstream of it. A possible splicing model explaining these data and involving formation of alterantive hairpin loop structures is proposed.     

利用~1H-NMR模拟谱研究三核苷酸A3′P(CH_3)5′A3′P(CH_3)5′A的构象性质

根据Altona等人的方法.利用~1H—NMR模拟谱分析了A3′P(CH_3)5A3′P(CH_3)5′A的三种非对映异构体:a、b和C的构象状态.发现:在室温下三个核糖环都是S型构象占优势(X_N≤0.44),而且随温度升高S型构象成分增多;两个二核苷酸片段-PAPA和APAP-的磷酸骨架扭角分别与A3′P(CH_3)5′A两种异构体a或b数值接近.差值约5°左右;二核苷酸片段中核糖环的重叠程度与手性磷原子的构型有关、R构型比S构型的核糖环重叠程度小;从而推测,A3′P(CH_3)5′A3′P(CH_3)5′A各种非对映异构体的稳定性为RR>RS.SR>SS,与化学合成中所见相符合.     

恐龙遗址旅游指南(3)欧洲篇(中A)

第二站:英国东苏塞克斯郡刘易斯镇(Lewes),曼特尔故居(Mantell’s house)其实这一站本应是我们这趟旅程的起点,虽然错过,但此处的光辉依然夺目,且将是永不消逝。恐龙,甚至整个古生物学科就在此起步。在此之前,人们从来未曾想过,在我们的后院里,在我们的家园上,居然有一群叫“恐     

Synthesis of 27-Oxo, 27-Hydroxymilbemycins A3 and A4 and Novel 27-Alkoxymilbemycins A3 and A4 from Milbemycins A3 and A4 and…

27-Oxomilbemycins A₃ and A₄ and 27-hydroxymilbemycins A₃ and A₄ were identified as metabolites in soil metabolism studies of milbemycins A₃ and A₄. Chemical derivation methods were developed to synthesize 27-oxomilbemycins A₃ and A₄ and 27-hydroxymilbemycins A₃ and A₄ from milbemycins A₃ and A₄. In addition, 27-alkoxymilbemycin derivatives were also synthesized from the same precursors. Some of the synthesized compounds displayed satisfactory acaricidal activity against the organophosphorus-sensitive two-spotted spider mite (Tetranychus urticae), but did not have superior activity to corresponding milbemycins A₃ and A₄.     

Phosphorylation and Inactivation of Glycogen Synthase Kinase 3β (GSK3β) by Dual-specificity Tyrosine Phosphorylation-regulated Kinase 1A (Dyrk1A)

Glycogen synthase kinase 3β (GSK3β) participates in many cellular processes, and its dysregulation has been implicated in a wide range of diseases such as obesity, type 2 diabetes, cancer, and Alzheimer disease. Inactivation of GSK3β by phosphorylation at specific residues is a primary mechanism by which this constitutively active kinase is controlled. However, the regulatory mechanism of GSK3β is not fully understood. Dual-specificity tyrosine phosphorylation-regulated kinase 1A (Dyrk1A) has multiple biological functions that occur as the result of phosphorylation of diverse proteins that are involved in metabolism, synaptic function, and neurodegeneration. Here we show that GSK3β directly interacts with and is phosphorylated by Dyrk1A. Dyrk1A-mediated phosphorylation at the Thr³⁵⁶ residue inhibits GSK3β activity. Dyrk1A transgenic (TG) mice are lean and resistant to diet-induced obesity because of reduced fat mass, which shows an inverse correlation with the effect of GSK3β on obesity. This result suggests a potential in vivo association between GSK3β and Dyrk1A regarding the mechanism underlying obesity. The level of Thr(P)³⁵⁶-GSK3β was higher in the white adipose tissue of Dyrk1A TG mice compared with control mice. GSK3β activity was differentially regulated by phosphorylation at different sites in adipose tissue depending on the type of diet the mice were fed. Furthermore, overexpression of Dyrk1A suppressed the expression of adipogenic proteins, including peroxisome proliferator-activated receptor γ, in 3T3-L1 cells and in young Dyrk1A TG mice fed a chow diet. Taken together, these results reveal a novel regulatory mechanism for GSK3β activity and indicate that overexpression of Dyrk1A may contribute to the obesity-resistant phenotype through phosphorylation and inactivation of GSK3β.     

真核细胞mRNA3′端poly(A)序列长度测定

在生物体中,信使核糖核酸(mRNA)的生物学功能是从DNA接受遗传信息,再以密码形式翻译出特异性蛋白质。mRNA对于基因表达和遗传信息传递起着重要作用。真核细胞mRNA分子的结构较为复杂,其特征之一是它的3′端几乎都有多聚腺嘌呤核苷酸序列     

α-Trifluoromethyl-β-Ureido-Propionic Acid (F3MUPA): A New Metabolite of Trifluridine (F3TdR)

Abstract

The metabolism of 5-trifluro-2′-deoxythymidine (trifluridine; F₃TdR) in male BALB/c mice bearing EMT-6 tumors has been investigated using ¹9F NMR spectroscopy. We previously (Tandon et al, 1992) reported the detection and identification of 5,6-dihydro-5-trifluorothymine (DHF₃T, 3) and 5,6-dihydroxy-5-trifluorothymine (DOHF₃T) as new metabolites of F₃TdR in mice urine. Further exploration of the metabolism of trifluridine has led to the identification of α-trifluoromethyl-β-ureido-propionic acid (F₃MUPA, 4) as a previously unreported metabolite in the urine of F₃TdR treated mice. Authentic F₃MUPA was obtained by synthesis via an established route. A comparison of chemical shift and the H-F coupling constant of an authentic sample, with the ¹⁹F signal from urine, indicated the presence of F₃MUPA in murine urine. Mixing crude urine with authentic F₃MUPA resulted in the enhancement of the corresponding fluorine signal without affecting or introducing others, thereby confirming the presence of F₃MUPA as a urinary metabolite.     

利用~1H-NMR模拟谱研究二核苷酸A3′P(CH_3)5′A的构象性质

本文根据Altona等人的方法,在利用~1H-NMR模拟谱确定18℃、40℃、70℃三种温度下有关质子化学位移及偶合常数的基础上,分析了A3′P(CH_3)5′A的两种非对映异构体a和b的构象状态。它们与A3′P(OH)5′A相比发现:(1)在18℃时两个核糖环是S型构象占主要成分,并随温度升高S型构象成分增多;(2)磷酸骨架的扭角ε′和β′分别改变±15°及±4°;(3)在优势构象情况下,两个核糖环都部分重叠,而且A3′P(CH_3)5′A(a)的重叠程度比A3P′(CH_3)5′A(b)小;(4)腺嘌呤碱基都更倾向去堆积状态,而且随温度升高A3′P(CH_3)5′A(a)比A3′P(CH_3)5′A(b)有着更强的去堆积效应。     

利用~1H-NMR模拟谱研究二核苷酸A3′P(CH_3)5′A的构象性质

本文根据Altona等人的方法,在利用~1H-NMR模拟谱确定18℃、40℃、70℃三种温度下有关质子化学位移及偶合常数的基础上,分析了A3′P(CH_3)5′A的两种非对映异构体a和b的构象状态。它们与A3′P(OH)5′A相比发现:(1)在18℃时两个核糖环是S型构象占主要成分,并随温度升高S型构象成分增多;(2)磷酸骨架的扭角ε′和β′分别改变±15°及±4°;(3)在优势构象情况下,两个核糖环都部分重叠,而且A3′P(CH_3)5′A(a)的重叠程度比A3P′(CH_3)5′A(b)小;(4)腺嘌呤碱基都更倾向去堆积状态,而且随温度升高A3′P(CH_3)5′A(a)比A3′P(CH_3)5′A(b)有着更强的去堆积效应。     

A deletion mutation in the βA1/A3 crystallin gene (CRYBA1/A3) is associated with autosomal dominant congenital nuclear cataract in a Chinese family

Congenital cataracts are an important cause of blindness worldwide. In a family of Chinese descent, a dominant congenital nuclear cataract locus was mapped to chromosome 17q11.1-12. The maximum LOD score, 2.49, at recombination fraction 0, was obtained for marker D17S1294. The results of both linkage and haplotype analyses defined a disease-gene to an 11.78-cM region harboring the gene coding for A1/A3 crystallin (CRYBA1/A3). Mutation analysis of the CRYBA1/A3 gene identified a 3-bp deletion in exon 4, which cosegregated with the disease risk in this family and was not observed in 100 normal chromosomes. This mutation resulted in the deletion of a highly conserved glycine at codon 91 (G91) and could be associated with an incorrect folding of A1/A3 crystallin. It highlights the physiological importance of crystallin and supports the role of CRYBA1/A3 in human cataracts formation.Y. Qi and H. Jia contributed equally to this work     

A20 (TNFAIP3) Alleviates CVB3-Induced Myocarditis via Inhibiting NF-κB Signaling

Background

Viral myocarditis, which is most prevalently caused by Coxsackievirus B3 (CVB3) infection, is a serious clinical condition characterized by cardiac inflammation. However, efficient therapies targeting inflammation are still lacking and much needed. A20, also known as tumor necrosis factor alpha induced protein 3 (TNFAIP3) is a key negative regulator of inflammation. But whether A20 may affect cardiac inflammation during acute viral myocarditis remains to be elucidated. The aim of this study was to investigate the potential protective effect of A20 on CVB3-induced myocarditis.

Methodology/Principal Findings

Mice were intraperitoneally inoculated with CVB3 to establish acute viral myocarditis model. We found that the expression of pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and monocyte chemotactic protein-1 (MCP-1) were markedly and persistently increased during the progression of CVB3-induced myocarditis, and positively correlated with the disease severity. Notably, intravenous injection in vivo with adenovirus expressed A20 (Ad-A20) remarkably reduced CVB3-induced pro-inflammatory cytokines production and alleviated the severity of myocarditis. Further, we observed that nuclear factor-kappaB (NF-κB) signaling which mediates inflammatory response was significantly inhibited in CVB3-infected mice with Ad-A20 treatment. Finally, we revealed that A20 was required to inhibit CVB3-induced NF-κB signaling by restricting TNF receptor associated factor 6 (TRAF6) ubiquitylation.

Conclusion/Significance

This study demonstrates the protective role of A20 against CVB3-induced myocarditis, which may provide a new therapeutic strategy for the treatment of viral myocarditis.     

A Short Syhthesis of 3′-(Methylsulfinyl)-3′-Deoxythymidine and Related Analogues

Abstract

The ring opening of the O-2,3′-anhydrothymidine 5 with the anion of methyl mercaptan gave the 3′-methylthio derivaative 6. Subsequent oxidation and deprotection afforded 3′-(methyl-sulfinyl)-3′-deoxythymidine 2 and its sulfone analogue 3.     

第11届国际生物奥林匹克竞赛(2000)理论试题A部分(3)

(续 2 0 0 1年第 7期第 44页 )植物形态和生理学60 .下面哪种未能阐明环式光合磷酸化 ?A.当细胞需要 ATP多于 NADPH更有利B.当 NADP 供应不足时更有利C.高能电子由铁氰化物首先接受D.质子菁是高能电子到达中心之前的最终受体E.在系统中 ,细胞色素 f把铁氧还蛋白和质体醌联系起来61.删除62 .1)对一些脱氢酶、脱羧酶、激酶、氧化酶和过氧化物酶的活性是需要的 ;2 )当缺乏时 ,植物组织变得柔软 ,且常常是软弱的 ,特别在低温和协迫的条件下 ;3 )对光合作用反应 ,包括 O2 的循环是需要的。选择哪种组合有利于上述内容 ?A.　 N　Ca　Mg　…     

3′-Oxo-tabernaelegantine A (OTNA) selectively relaxes pulmonary arteries by inhibiting AhR

BackgroundPulmonary arterial hypertension (PAH), characterized by pulmonary artery constriction and vascular remodeling, has a high mortality rate. New drugs for the treatment of PAH urgently need to be developed.PurposeThis study was designed to investigate the vasorelaxant activity of OTNA in isolated pulmonary arteries, and explore its molecular mechanism.MethodsPulmonary arteries and thoracic aortas were isolated from mice, and vascular tone was tested with a Wire Myograph System. Nitric oxide levels were determined with DAF-FM DA and DAX-J2™ Red. Cellular thermal shift assays, microscale thermophoresis, and molecular docking were used to identify the interaction between OTNA and aryl hydrocarbon receptor (AhR). The levels of PI3K, p-PI3K, Akt, p-Akt, eNOS, p-eNOS, and AhR were analyzed by Western blotting.ResultsOTNA selectively relaxed the isolated pulmonary artery rings in an endothelium-dependent manner. Mechanistic study showed that OTNA induced NO production through activation of the PI3K/Akt/eNOS pathway in endothelial cells. Furthermore, we also found that OTNA directly bound to AhR and activated the PI3K/Akt/eNOS pathway to dilate pulmonary arteries by inhibiting AhR.ConclusionsOTNA relaxes pulmonary arteries by antagonizing AhR. This study provides a new natural antagonist of AhR as a promising lead compound for PAH treatment.     

Possibility Theory: A New Approach to Uncertainty Analysis? (3 pp)

Background, Aims and Scope The problem of the evaluation of practitioner's belief and belief-related uncertainties on LCA results obtained from different methodological choices has been addressed so far by scenario modeling, Cultural Theory perspectives and probabilistic simulation. The direct evaluation of belief and related uncertainties could be of interest, e.g. when the information available (resulting from classical uncertainty analysis or the application of the precautionary principle) do not allow one to choose between methodological alternatives leading to different LCA results and conclusions. The difficulty of modeling belief arises from the additive nature of classical measures, e.g. probabilities. Since the 1960s, non-additive measures (e.g. possibilities) have been developed and applied to model belief in real world problems. The aim of this paper is to discuss the application of possibility measures in LCA for uncertainty analysis in complement to classical approaches. Methods The nature and the meaning of possibilities are briefly introduced by comparison with probabilities (subjective or not) in order to enlighten strengths, drawbacks and complementarities. A tentative possibilistic approach based on the evaluation of a posteriori possibilities of final LCA results depending on a priori possibilities of the methodological choices behind the calculations is described, also by means of an application example. Results and Outlook. A new approach for the modeling of practitioner's belief and belief-related uncertainties in complement of classical methods of uncertainty analysis has been proposed for discussion. Uncertainty can be characterized by confidence intervals and indexes that could help practitioners in making methodological choices and could improve the interpretation and reliability of LCA results, still increasing its sophistication.     

细胞外钙调素诱导番茄悬浮细胞rbcS-3A基因表达(英文)

利用番茄 (LycopersicumesculentumMill.)悬浮培养细胞为材料 ,以3 2 P标记的寡核苷酸 (40bp)为探针检测了细胞外钙调素对番茄细胞rbcS_3A及rbcS_3C基因表达的影响。当向暗中培养的番茄悬浮细胞 (第 7天 )中加入外源纯化钙调素 (10 -7mol/L)并处理 2 4h后 ,rbcS_3A基因的表达明显增加 ,而相同浓度的S_10 0蛋白和BSA则没有作用。加入外源钙调素 (10 -7mol/L)对rbcS_3C基因的表达没有影响。上述结果表明细胞外钙调素对暗中培养的番茄rbcS基因的表达有调控作用 ,并且这种调控作用具有亚型特异性 ,即对番茄rbcS_3A基因的表达有诱导作用 ,而对番茄rbcS_3C基因的表达没有作用。     

Tumor Necrosis Factor (TNF) –308G>A,Nitric Oxide Synthase 3 (NOS3) +894G>T Polymorphisms and Migraine Risk: A Meta-Analysis

Background and Objective

Conflicting data have been reported on the association between tumor necrosis factor (TNF) –308G>A and nitric oxide synthase 3 (NOS3) +894G>T polymorphisms and migraine. We performed a meta-analysis of case-control studies to evaluate whether the TNF –308G>A and NOS3 +894G>T polymorphisms confer genetic susceptibility to migraine.

Method

We performed an updated meta-analysis for TNF –308G>A and a meta-analysis for NOS3 +894G>T based on studies published up to July 2014. We calculated study specific odds ratios (OR) and 95% confidence intervals (95% CI) assuming allele contrast, dominant model, recessive model, and co-dominant model as pooled effect estimates.

Results

Eleven studies in 6682 migraineurs and 22591 controls for TNF –308G>A and six studies in 1055 migraineurs and 877 controls for NOS3 +894G>T were included in the analysis. Neither indicated overall associations between gene polymorphisms and migraine risk. Subgroup analyses suggested that the “A” allele of the TNF –308G>A variant increases the risk of migraine among non-Caucasians (dominant model: pooled OR = 1.82; 95% CI 1.15 – 2.87). The risk of migraine with aura (MA) was increased among both Caucasians and non-Caucasians. Subgroup analyses suggested that the “T” allele of the NOS3 +894G>T variant increases the risk of migraine among non-Caucasians (co-dominant model: pooled OR = 2.10; 95% CI 1.14 – 3.88).

Conclusions

Our findings appear to support the hypothesis that the TNF –308G>A polymorphism may act as a genetic susceptibility factor for migraine among non-Caucasians and that the NOS3 +894G>T polymorphism may modulate the risk of migraine among non-Caucasians.     

DF-6 P3A型超声波粉碎机 DF-6P3A型智能超声波粉碎机

简讯 宁波师范学院实验总厂是浙江省教委、宁波市教委下属研究、生产生化仪器和教学仪器的科技实体,技术力量雄厚,质量检验手段先进,产品结构合理,性能可靠,并取得省科技部门和省教仪公司鉴定。该厂生产的超声波粉碎机是利用强超声在液体中产生空化效应原理,对生物材料等物质进行超声处理的多功能、多用途仪器,用于粉碎细胞、乳化、分散、提取、匀化、消泡、清洗及加速化学反应等,是生物、化学、农林、医药、食品、环保、卫生、能源等领域的得力工具。目前可供施种类型的超声波粉碎机。