酵母校正tRNA的提取及其对番茄花叶病毒增殖的抑制作用

从酵母(Saccharamyces cerevisiae SUP-6-1温度突变株,含SUPt RNATyr,SUFtRNAleo,SUPtRNAHis)和菠菜提取了总tRNA,经过BD-纤维素柱层析,得到部份纯化的amber终止密码校正tRNA,它们在兔网织细胞裂解液翻译体系中能促进番茄花叶病毒(ToMV)RNA183k抄读蛋白的合成。研究了酵母SUP6-l校正tRNA对ToMV在烟草中增殖的影响。在用酵母SUP6-l校正tRNA处理的植株顶部新生叶中病毒滴度接种3、5、11、15和20天后,分别是对照的3％、12％、3 8％、42％和67％。用校正tRNA处理的烟草植株中层叶片,在接种后11—20天明显低于对照。下层接种叶中的病毒滴度无显著差异。讨论了校正tRNA对ToMV增殖抑制的机理。     

感染番茄花叶病毒及其弱毒系的烟叶中病毒双链RNA的比较

利用3H-尿嘧畦核苷标记技术、抗Rnase性质和聚丙烯酰胺凝肢电泳分析,比较了接种番茄花叶病毒(ToMV)及其弱毒系N14的烟叶中ds-RNA含量的变化。接种后第2天,N14的抗Rnase的ds-RNA为58．4％,ToMV为4．8％随着接种时间延长,ToMV的ds一RNA迅速降低到10.4—18．5％,而N14 的仍保持在25．5--36．6％,的较高水平。Ds一RNA的聚丙烯酰胺凝肢电泳分析证实了上述结果。讨论了ds-RNA在具有单链RNA为基因组的病毒毒力中的可能作用,ds-RNA含量高可能降低病毒合成能力和／或诱导某种类似干扰素的抗病毒物质的产生。     

高温胁迫下番茄幼苗对冷激的响应

高温逆境是影响夏秋季蔬菜设施集约化育苗质量的主要因素之一,利用温度逆境诱导植物产生交叉适应是植物获得抗逆性的一种有效手段.为探索冷激强度对番茄幼苗高温胁迫的缓解效应,试验采用人工气候箱模拟夏季设施中的高温胁迫,研究了不同冷激温度(5、10、15 ℃)和冷激持续时间(10、20、30 min)对番茄幼苗生长、生物膜保护系统的影响,并研究了单次适宜冷激处理对番茄小分子热激蛋白LeHSP23.8和CaHSP18基因表达的影响.结果表明： 在高温胁迫前对番茄幼苗进行冷激处理可以抑制其下胚轴的伸长和株高的生长.冷激缓解番茄幼苗高温胁迫的效应在不同冷激温度下表现出不同的变化趋势;5 ℃冷激处理抑制了番茄幼苗抗氧化酶活性的升高,使细胞膜透性增大,对幼苗产生伤害;10 ℃冷激处理对番茄幼苗高温胁迫的缓解效应随冷激时间的延长呈降低趋势;而15 ℃冷激处理缓解番茄幼苗高温胁迫的效应随冷激时间的延长呈增加趋势.适宜冷激温度和冷激持续时间能够诱导番茄幼苗对高温逆境的交叉适应性,在高温胁迫前将番茄幼苗进行温度为10 ℃、持续10 min的冷激处理效果最佳,与对照相比,显著提高了高温胁迫下番茄幼苗植株单株干质量和壮苗指数,降低了番茄幼苗叶片相对电导率和丙二醛含量,促进了脯氨酸和可溶性蛋白的积累,提高了番茄幼苗叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT) 3种抗氧化酶活性,并诱导了小分子热激蛋白基因LeHSP23.8和CaHSP18在常温条件下的上调表达.
     

番茄凝集素的分离纯化及某些性质的研究

番茄成熟果实经抽提上鸡卵类粘蛋白(OM)-Sepharose 4B亲和层析柱分离纯化制得番茄凝集素。制品在SDS—聚丙烯酰胺凝胶电泳上呈一条带,表观分子量为205,000。Sephadex G200凝胶过滤行为呈单一吸收峰,分子量为180,000。等电聚焦测得等电点为7.6和9.4。它能使人和多种动物红细胞,以及某些培养细胞凝集,但效价不同。N—乙酰葡萄糖胺寡聚糖是其专一性结合糖。     

番茄花叶病毒移动蛋白和番茄抗病基因Tm-2^2的互作诱导烟草转化体程序性细胞死亡

番茄的抗病基因Tm -2 2 与番茄花叶病毒 (ToMV)的移动蛋白MP基因是一对互作的基因 ,Tm- 2 2 基因和ToMV MP基因同时在烟草中表达 ,并分别获得单一基因整合的纯合转化体植株。病毒接种试验表明 ,Tm -2 2 基因转化体与Tm- 2 2 番茄对Tobamavirus病毒的特异抗性结果一致 ;Tm -2 2 转基因植株和ToMV MP转基因植株杂交试验及其农杆菌注射试验均证明 :(1)Tm -2 2 基因与ToMV- MP在转基因烟草上保持“基因对基因”的互作关系 ;(2 )在外源乙烯的参与下 ,ToMV的移动蛋白与Tm -2 2 基因编码蛋白的互作能够诱导转化体程序性细胞死亡。这一结果为今后研究Tm -2 2 与MP互作的分子机制奠定了基础。     

叶面喷施亚精胺对高温胁迫下番茄叶绿素合成代谢的影响

以设施番茄栽培品种‘金棚朝冠’幼苗为试验材料,研究叶面喷施0.25 mmol·L~(-1)亚精胺(Spd)对高温胁迫下(38℃/28℃,昼/夜)番茄幼苗生长及叶绿素合成过程中前体物质含量、关键酶活性和叶绿素含量的影响,探讨Spd缓解番茄高温胁迫伤害的生理机制。结果表明:(1)高温胁迫显著抑制了番茄幼苗的生长,叶面喷施Spd可有效缓解高温胁迫对番茄幼苗地上部生长的抑制作用,但对于地下部的生长无显著缓解作用。(2)高温胁迫下番茄叶片叶绿素合成前体物质δ-氨基酮戊酸(ALA)和胆色素原(PBG)的含量显著提高,而尿卟啉原Ⅲ(UroⅢ)、原卟啉Ⅸ(ProtoⅨ)、镁-原卟啉Ⅸ(Mg-protoⅨ)和原叶绿素酸(Pchl)的含量显著降低,证明叶绿素前体由PBG向UroⅢ的转化受阻,导致叶绿素a(Chla)、总叶绿素(Chlt)含量和Chla/Chlb显著降低。(3)叶面喷施Spd能增强高温胁迫下胆色素原脱氨酶(PBGD)活性,有效缓解了高温胁迫对PBG到UroⅢ转化的阻碍作用,促进PBG之后叶绿素合成前体物质的合成,Chla含量和Chla/Chlb显著增加。研究发现,高温胁迫显著抑制番茄幼苗的生长,叶面喷施Spd可通过显著增强PBGD活性来缓解由PBG向UroⅢ的受阻程度,促进高温胁迫下番茄叶片的叶绿素前体合成,提高叶绿素含量和番茄植株的生长量,减轻高温胁迫对番茄幼苗生长的伤害。     

烟草花叶病毒和番茄花叶病毒在含N基因烟草上的症状差异是由运动蛋白基因决定的

烟草花叶病毒(TMV)和番茄花叶病毒(ToMV)是烟草花叶病毒属中关系最为密切的病毒, 但它们在含N基因烟草上产生的枯斑大小有明显的差异. 比较了TMV, ToMV及用ToMV运动蛋白基因(MP)精确置换TMV MP后获得的重组病毒T/OMP在不同寄主上的症状差异, 发现T/OMP在含N基因烟草上产生的枯斑大小与ToMV相似. 分析比较TMV, ToMV和T/OMP外壳蛋白和MP在植物体内的积累水平, 发现三者之间没有明显的差异, 而TMV和T/OMP在原生质体中的复制水平也没有差异. 比较TMV, ToMV和T/OMP接种后烟草体内防御相关酶(PAL, POD和PPO)的活性变化, 结果T/OMP和TMV所诱导酶的变化趋势基本一致, 而与ToMV有所差异, 因此认为MP基因功能的差异决定了TMV和ToMV在N基因烟草上枯斑的大小.     

二斑叶螨为害对番茄叶片主要营养物质和防御酶活性的影响

本文研究了二斑叶螨Tetranychus urticae为害对寄主植物番茄营养物质和防御酶活性的影响。结果表明:二斑叶螨为害对番茄植株体内营养物质含量和防御酶活性均有一定影响,且为害密度和为害时间存在交互作用。二斑叶螨为害后番茄叶片可溶性糖含量下降幅度与叶螨为害密度和时间呈正比;可溶性蛋白和游离氨基酸含量在二斑叶螨高密度为害3 d时最低。二斑叶螨为害后番茄叶片防御酶活性普遍增强,过氧化物酶POD活性在5头/叶密度时与为害时间呈正比;苯丙氨酸解氨酶PAL、多酚氧化酶PPO和脂氧合酶LOX活性的变化一致,均在中密度为害时活性最高。以上结果说明番茄叶片通过调节其体内营养物质含量和各种防御酶活性的变化,对二斑叶螨的胁迫产生了应激反应。     

一种利用普通垂直电泳槽回收PAGE胶蛋白条带的简便方法

植物总蛋白样品经聚丙烯酰胺凝胶电泳分离之后,直接用考马斯亮蓝染色、切胶回收目的条带,再用聚丙烯酰胺凝胶电泳槽电洗脱纯化得到单一条带的目的蛋白.此法可得到有活性的黄瓜衰老叶片中被特异激活的DNA酶,对样品中含量少,特别是与其他分子量相近的蛋白质十分有效.     

夜间低温对番茄幼苗磷素吸收及转运的影响

以‘辽园多丽’番茄幼苗为试材,采用营养液栽培模式,以夜温15℃为对照,对夜间低温(6℃)影响番茄幼苗磷素吸收及转运过程的因素进行研究。结果显示:(1)夜间低温胁迫导致番茄幼苗根系活力受到显著抑制。(2)夜间低温条件下,番茄幼苗根系中酸性磷酸酶活性无明显变化,而其地上部酸性磷酸酶活性增强,且以叶片中活性增加较大。(3)夜间低温胁迫使根系中磷酸盐转运蛋白基因LePT1和LePT2的相对表达量较对照增加,地上部磷酸盐转运蛋白基因LePT1的表达受到抑制,且叶片中受到的抑制作用更显著。(4)夜间低温胁迫处理营养液中剩余磷素含量始终多于对照;其番茄幼苗地下部和地上部中磷素绝对含量均下降,且叶片比茎的下降幅度更大。(5)夜间低温胁迫使番茄幼苗伤流强度下降,伤流液中磷素含量随着处理天数的增加而增加,在处理第9天时极显著高于对照。研究表明,夜间低温导致番茄幼苗根系活力降低,诱导植株中磷酸盐转运蛋白基因LePT1的表达下调以及伤流强度降低,从而引起磷素由茎向叶片中的转运过程受到明显抑制。     

Effect of self-assembly on antimicrobial activity of double-chain short cationic lipopeptides

Short cationic antimicrobial lipopeptides with surfactant-like structure are promising antibiotic candidates that preferentially target microbial membranes. Therefore, we focused our study on double-chain lipopeptides, (C_10-16)₂Dab-KKK-NH₂ and (C_10-16)₂Dap-KKK-NH₂, where Dab and Dap are 2,4-diaminobutyric and 2,3-diaminopropionic acids, respectively. We tried to answer a question how the self-assembly behaviour affects biological activities of the tested compounds. The subject compounds were synthesized by solid-phase method and screened for their antimicrobial and haemolytic activities. Cytotoxicity tests on human keratinocytes were carried out for the most promising lipopeptides. Self-assembly properties were evaluated by both experimental and theoretical methods. Interactions with membrane models were examined using the ITC and FTIR techniques. All the lipopeptides studied showed the tendency to self-assembly in solution, and this behaviour was affected by the length of the hydrocarbon chains. Acyl chain elongation supported the formation of the bilayer structure and deprived the lipopeptides of antimicrobial activity. A multi-step mechanism of interaction with a negatively charged membrane was observed for the short-chain lipopeptides, indicating other processes accompanying the binding process. Short-chain lipopeptides were able to penetrate into the liposome’s interior and/or cause the rupture of the liposome, this being compatible with their high antimicrobial activity.     

Transposition burst of mariner-like elements in the sequenced genome of Rhodnius prolixus

Transposable elements (TEs) are widespread in insect's genomes. However, there are wide differences in the proportion of the total DNA content occupied by these repetitive sequences in different species. We have analyzed the TEs present in R. prolixus (vector of the Chagas disease) and showed that 3.0% of this genome is occupied by Class II TEs, belonging mainly to the Tc1-mariner superfamily (1.65%) and MITEs (1.84%). Interestingly, most of this genomic content is due to the expansion of two subfamilies belonging to: irritans himar, a well characterized subfamily of mariners, and prolixus1, one of the two novel subfamilies here described. The high amount of sequences in these subfamilies suggests that bursts of transposition occurred during the life cycle of this family. In an attempt to characterize these elements, we performed an in silico analysis of the sequences corresponding to the DDD/E domain of the transposase gene. We performed an evolutionary analysis including network and Bayesian coalescent-based methods in order to infer the dynamics of the amplification, as well as to estimate the time of the bursts identified in these subfamilies. Given our data, we hypothesized that the TE expansions occurred around the time of speciation of R. prolixus around 1.4 mya. This suggestion lays on the “Transposon Model” of TE evolution, in which the members of a TE population that are replicative active are present at multiple loci in the genome, but their replicative potential varies, and of the “Life Cycle Model” that states that when present-day TEs have been involved in amplification bursts, they share an ancestral copy that dates back to this initial amplification.     

Potential chemical transformation of phosphinic acid derivatives and their applications in the synthesis of drugs

The chemical transformation of phosphinic acid is a well-considered mature area of research on account of the historical significant reactions such as Kabachnik–Fields, Mannich, Arbuzov, Michaelis–Becker, etc. Considerable advances have been made over last years especially in metal-catalyzed, free-radical processes and asymmetric synthesis using catalytic enantioselective. As a result, the aim of this synopsis is to make the reader familiar with advances in the approaches of phosphinic acids toward the synthesis of highly functionalized and valuable buildings blocks. Another purpose of this survey is to provide the current status of the applications of phosphinic acids in the synthesis of drugs.     

Synthesis and biological evaluation of aryl phosphoramidate prodrugs of fosfoxacin and its derivatives

Aryl phosphoramidate prodrugs of fosfoxacin derivatives 15a-b and 8a-b were synthesized and investigated for their ability to target bacteria. No growth inhibition was observed neither for Mycobacterium smegmatis nor for Escherichia coli on solid medium, demonstrating the absence of release of the active compounds in the bacterial cells. Investigation of the stability of the prodrugs and their multienzymatic cleavage in abiotic and biotic conditions showed that the use of aryl phosphoramidate prodrug approach to deliver non-nucleotides compounds is not obvious and might not be appropriate for an antimicrobial drug.     

Therapeutic utility of the newly discovered properties of interleukin-21

Since its discovery in 2000, interleukin-21 (IL-21) has been shown to display a broad spectrum of pleiotropic actions including the regulation of development, differentiation and function of lymphoid-myeloid cells. More specifically, IL-21 modulates the effector functions of T, B and NK cells, which not only have key roles in antitumoral and antiviral immunity but also in exerting major effects on inflammatory responses promoting the development of autoimmune diseases. Recent studies have unveiled an unexpected role for IL-21 in immune regulation and de novo T-cell development. While highlighting its critical role in immunity, this review will mainly focus on recent advances in IL-21 biology and how such newly discovered properties could potentially be exploited therapeutically in the establishment of future clinical trials.     

Lipids of mitochondria

A unique organelle for studying membrane biochemistry is the mitochondrion whose functionality depends on a coordinated supply of proteins and lipids. Mitochondria are capable of synthesizing several lipids autonomously such as phosphatidylglycerol, cardiolipin and in part phosphatidylethanolamine, phosphatidic acid and CDP-diacylglycerol. Other mitochondrial membrane lipids such as phosphatidylcholine, phosphatidylserine, phosphatidylinositol, sterols and sphingolipids have to be imported. The mitochondrial lipid composition, the biosynthesis and the import of mitochondrial lipids as well as the regulation of these processes will be main issues of this review article. Furthermore, interactions of lipids and mitochondrial proteins which are highly important for various mitochondrial processes will be discussed. Malfunction or loss of enzymes involved in mitochondrial phospholipid biosynthesis lead to dysfunction of cell respiration, affect the assembly and stability of the mitochondrial protein import machinery and cause abnormal mitochondrial morphology or even lethality. Molecular aspects of these processes as well as diseases related to defects in the formation of mitochondrial membranes will be described.     

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/.     

Role of antigen presentation in the production of pro-inflammatory cytokines in obese adipose tissue

Type II diabetes regroups different physiological anomalies that ultimately lead to low-grade chronic inflammation, insulin resistance and loss of pancreatic β-cells. Obesity is one of the best examples of such a condition that can develop into Metabolic Syndrome, causing serious health problems of great socio-economic consequences. The pathological outcome of obesity has a genetic basis and depends on the delicate balance between pro- and anti-inflammatory effectors of the immune system. The causal link between obesity and inflammation is well established. While innate immunity plays a key role in the development of a pro-inflammatory state in obese adipose tissues, it has now become clear that adaptive immune cells are also involved and participate in the cascade of events that lead to metabolic perturbations. The efficacy of some immunotherapeutic protocols in reducing the symptoms of obesity-driven metabolic syndrome in mice implicated all arms of the immune response. Recently, the production of pathogenic immunoglobulins and pro-inflammatory cytokines by B and T lymphocytes suggested an auto-immune basis for the establishment of a non-healthy obese state. Understanding the cellular landscape of obese adipose tissues and how immune cells sustain chronic inflammation holds the key to the development of targeted therapies. In this review, we emphasize the role of antigen-presenting cells and MHC molecules in obese adipose tissue and the general contribution of the adaptive arm of the immune system in inflammation-induced insulin resistance.     

Insights of synthetic analogues of anti-leprosy agents

Today, the emergence of the phenomenon of drug or multidrug-resistance for community-associated diseases represents a major concern in the world. In these contexts, the chronic infectious disease, leprosy, grounded by a slow-growing bacterium called Mycobacterium leprae or Mycobacterium lepromatosis is a leading cause of severe disfiguring skin sores and nerve damage in the arms, legs, and skin areas around the body. Even, over 200,000 new leprosy cases are being accounted every year along with the relapsed leprosy cases. Nonetheless, this has been considered a curable disease with a higher dose of multidrug therapy (MDT) for a long period of time. The prolonged action of a high dose of combination drugs administration may cause an adverse reaction that can significantly affect patient compliance, particularly the outbreak of multidrug-resistance in the infected person. To overcome these shortfalls or prevent the resistance-associated problems, researchers are diligently involved in the structural modifications of the clinically used anti-leprosy drugs or the allied compounds for the structure-antimycobacterial activity relationship study. This review article described the detailed synthesis and biological assays of different anti-leprosy compounds reported by several research groups.