A plasmid-encoded arsenite pump produces arsenite resistance in Escherichia coli

The arsenate resistance operon of R-factor R773, a conjugative resistance plasmid, has two functional regions, a promoter-proximal region encoding resistance to arsenite and antimonate, and a promoter-distal one encoding arsenate resistance. Cells bearing arsenite resistance plasmids exhibited reduced accumulation of 74AsO2-. When resistant cells were depleted of endogenous energy reserves and then loaded with 74AsO2-, active extrusion of the ion was observed when an energy source was supplied. Intracellular ATP was required for extrusion, but a proton motive force was neither necessary nor sufficient. An arsenite-sensitive mutant was unable to extrude arsenite, while an arsenate-sensitive mutant had normal arsenite transport. These results suggest that the action of a plasmid-encoded primary arsenite efflux pump is the mechanism of arsenite resistance.     

Role of residual Sb(III) in meglumine antimoniate cytotoxicity and MRP1-mediated resistance

Despite the clinical use of pentavalent antimonials for more than half a century, their metabolism in mammals and mechanisms of action and toxicity remain poorly understood. It has been proposed that the more active and toxic trivalent antimony form Sb(III) plays a critical role in their antileishmanial activity and toxicity. The aim of this work was to investigate the role of residual Sb(III) both in the antileishmanial/antitumoral activities of the pentavalent meglumine antimoniate and in the MRP1 (multidrug resistance-associated protein 1)-mediated resistance to this drug. Samples of meglumine antimoniate differing in their amount of residual Sb(III) (meglumine antimoniate synthesized either from SbCl₅ or from KSb(OH)₆ as well as commercially-available meglumine antimoniate) were evaluated in vitro and in vivo on Leishmania amazonensis infections, as well as for their cytotoxicity to normal and MRP1-overexpressing GLC4 cell lines. Although in vitro the two most effective drugs contained the highest levels of Sb(III), no correlation was found in vivo between the antileishmanial activity of meglumine antimoniate and its residual Sb(III) content, suggesting that residual Sb(III) contributes only marginally to the drug antileishmanial activity. On the other hand, the GLC4 cells growth inhibition data strongly suggests a marked contribution of residual Sb(III). Additionally, the potassium salt of antimoniate (non-complexed form of Sb(V)) was found to be more cytotoxic than meglumine antimoniate. Although MRP1-overexpressing GLC4 cells showed a marked resistance to trivalent antimonials, cross-resistance to meglumine antimoniate was observed only for the products that contained relatively high levels of Sb(III) (at least 0.03% by weight), suggesting that MRP1 mediates resistance to Sb(III) but not to Sb(V). In conclusion, our data strongly suggest that residual Sb(III) in pentavalent antimonial drugs does not contribute significantly to their antileishmanial activity, but is responsible for their cytotoxic activity against mammalian cells and the MRP1-mediated resistance to these drugs.     

New insights into the chemical structure and composition of the pentavalent antimonial drugs, meglumine antimonate and sodium stibogluconate

The chemical structures of the pentavalent antimonial drugs, meglumine antimonate (MA) and sodium stibogluconate (SSG), were re-evaluated using electrospray ionization mass spectrometry (ESI-MS) and osmolarity measurements. Both MA and SSG were found to contain 1:1, 1:2, 2:2 and 2:3 Sb(V)-ligand complexes. ESI-MS analysis of MA showed negatively-charged 1:1 (m/z 364) and 2:2 (m/z 765) Sb(V)-meglumine complexes, supporting the predominance of zwitterionic species in solution. Our data are consistent with a structure for the 1:2 Sb(V)-meglumine, which differs from that previously postulated, with two positively-charged amino groups and one negatively-charged antimonate group. Instead of the commonly hypothesized structure for SSG, in which two Sb atoms are linked by an oxygen, an alternative structure is proposed, based on the ability of Sb(V)-gluconate complexes to polymerize. MA (or SSG) in concentrated aqueous solutions, such as of MA (or SSG) in its commercial form, is expected to consist mainly of a mixture of 2:2, 2:3 and 2:1 Sb(V)-ligand complexes, as suggested by the 2:1 Sb-to-particle ratio found by osmometry. 1:1 Sb(V)-ligand complexes in MA and SSG are expected to play an important pharmacological role, as suggested by the slow increase of osmolarity of MA solution upon dilution at 37 degrees C (half-time of 20min).     

左旋卡尼汀联合环磷腺苷葡胺治疗慢性充血性心力衰竭

目的:探究左旋卡尼汀联合环磷腺苷葡胺治疗慢性充血性心力衰竭(chronic congestive heart failure,CHF)的临床疗效。方法:选取我院心内科CHF患者58例,随机分为对照组28例与实验组30例,其中对照组28例,予地高辛、氢氯噻嗪、环磷腺苷葡胺常规治疗;实验组30例,在常规治疗的基础上加用左旋卡尼汀治疗。观察两组患者治疗前后心功能分级(NYHA)、6 min步行距离、左心室射血分数(LVEF)及左心室舒张末容积(LVEDD)等。结果:与对照组比较,实验组患者治疗后心率和NYHA心功能分级明显改善,6 min步行实验明显提高,患者LVEDD、LVEF明显改善,差异具有统计学意义(P<0.05);两组患者接受相应治疗后均未出现任何不良反应(P>0.05)。结论:左旋卡尼汀联合环磷腺苷葡胺能够明显改善慢性充血性心力衰竭,对临床具有指导意义,值得临床推广。     

Testing of drugs for antileishmanial activity in golden hamsters infected with Leishmania donovani

Hanson W. L., Chapman W. L. Jr. and Kinnamon K. E. 1977. Testing of drugs for antileishmanial activity in golden hamsters infected with Leishmania donovani. International Journal for Parasitology7: 443–447.An existing procedure has been modified to make possible the screening of 10–15 selected compounds per week for suppressive activity against Leishmania donovani in a golden hamster model. Compounds with previously known antileishmanial activity were consistently found to have suppressive activity with this procedure. Antileishmanial activity is reported for the first time with compounds of the 6- and 7-aminoquinoline classes. The activity of all compounds studied is compared to that of Glucantime® which is the reference compound used in this procedure.     

热胁迫对辣椒花药发育过程中Ca2+分布的影响

运用焦锑酸钾沉淀法,研究了不同热胁迫时间对辣椒（Capsicum annuum L.)小孢子发生和花粉发育过程中Ca^2 分布的影响。在小孢子母细胞中,细胞表面有少量Ca^2 分布,细胞核中基本上观察不到Ca^2 ,热胁迫12h后,细胞质和细胞核中Ca^2 明显增多,液泡膜内侧也有许多Ca^2 分布,热胁迫24h后,大量的Ca^2 分布在细胞质中,液泡膜上和液泡内;在四分体时期,与小孢子母细胞相比,四分小孢子表面和细胞质中Ca^2 数量明显增加,热胁迫24h后,细胞质和细胞核中Ca^2 更多;在小孢子中,大量Ca^2 分布在壁上,质膜内侧,液泡膜上,少量分布在细胞质和细胞核中,热胁迫12h后,质膜上Ca^2 增多,24h后,细胞质,细胞核中,质膜内侧的Ca^2 继续增多,热胁迫对成熟花粉中Ca^2 的分布无明显影响。     

环磷腺苷葡胺联合培哚普利在慢性心力衰竭中的应用研究

目的:分析环磷腺苷葡胺联合培哚普利在慢性心力衰竭患者的应用价值。方法:以2013年1月至2015年12月新疆医科大学第五附属医院心血管内科诊治的慢性心力衰竭患者90例为研究对象,将其按照就诊病历顺序号分对照组、培哚普利组和联合组,每组30例。对照组患者接受常规治疗,培哚普利组患者在对照组的基础之上加用培哚普利,联合组患者在培哚普利组的基础之上加用环磷腺苷葡胺,三组患者均连续治疗2周,比较三组临床疗效及治疗前后患者的左心室重塑情况(左心室收缩末期内径(LVESD)和舒张末期内径(LVEDD))及左心室心功能(左心室射血分数(LVEF)),患者活动能力(6 min的步行距离)以及血浆脑钠利肽(BNP)、胰岛素样生长因子1(IGF-1)、胱抑素(Cys-C)水平。结果:经过2周的治疗,培哚普利组和联合组的治疗总有效率分别为86.67%和96.67%,均显著高于对照组的66.67%(P0.05),但培哚普利组和联合组比较差异无统计学意义(P0.05);三组患者的LVEDD、LVESD、血浆BNP水平均较治疗前显著降低,且培哚普利组和联合组均显著低于对照组(P0.05),联合组LVESD、血浆BNP水平显著低于培哚普利组;三组患者的LVEF和6 min步行距离、血浆IGF-1水平均较治疗前显著升高,且培哚普利组和联合组均显著高于对照组,联合组血浆IGF-1水平、LVEF和6 min步行距离显著高于培哚普利组(P0.05);联合组和培哚普利组治疗后的Cys-C水平治疗前显著降低,且联合组和培哚普利组显著低于对照组,联合组血Cys-C水平较培哚普利组更低(P0.05)。结论:环磷腺苷葡胺联合培哚普利治疗慢性心力衰竭临床疗效显著,可较单药治疗显著改善患者左心室重构情况、心室射血功能和活动能力,可能与协同调节患者血浆BNP、IGF-1、Cys-C水平有关。     

钙离子在水霉(Saprolegnia ferax)菌丝顶端细胞壁中呈极性分布

焦锑酸钾处理的水霉(Saprolegnia ferax)菌丝显示:焦锑酸沉淀颗粒仅存在菌丝细胞壁而不是原生质中,并呈顶端到基部的极性分布,即在菌丝最顶端细胞壁中丰富致密、重叠在一起,在约3—10μm的亚顶端则变得稍为稀薄而可分辨,在约10μm以后的成熟区域进一步显得松散而无规律。焦锑酸沉淀颗粒可经EGTA螫合处理去除,并经X-射线微区分析证明在3.6—3.7keV区域可产生Sb和Ca元素混合峰,说明焦锑酸沉淀反映了Ca~(2 )的分布。对冷冻干燥菌丝细胞壁表面进行扫描电镜X-射线微区分析,同样证明菌丝细胞壁含有大量的Ca,菌丝最顶端Ca信号强度高于10μm以后的成熟区。由于Ca~(2 )和H~ 可能为一对拮抗因子维持菌丝顶端细胞壁可塑性和刚性间的平衡以保证菌丝顶端生长,我们检验了菌丝生长过程中培养介质的pH变化,证实培养介质pH随培养时间的延长而逐渐下降。上述结果提示细胞壁Ca~(2 )可能在菌丝顶端生长过程中起作用。     

Meglumine as a green,efficient and reusable catalyst for synthesis and molecular docking studies of bis(indolyl)methanes as antioxidant agents

An efficient and convenient Meglumine catalyzed procedure for the synthesis of bis(indolyl) methanes at ambient temperature under aqueous conditions in high yields. The catalytic reaction proceeds very smoothly. Clean reaction, ease of product isolation/purification, easily available reactants, metal free and environmentally friendly reaction conditions are the notable advantages of the present methodology. All the entitled compounds were characterized by IR, ¹H, ¹³C NMR, mass spectra and evaluated for their antioxidant (DPPH, H₂O₂ and NO scavenging methods). They exhibited potent in vitro antioxidant activity dose-dependently. The binding interactions and molecular docking studies for entitled compounds were studied against 3MNG protein. 4d exhibited marked binding affinity with excellent docking score of −7.6 K.cal/mol and emerged as a lead compound.     

烟草花粉萌发和花粉管生长期间柱头和花柱中的钙分布

烟草柱头表面有两层覆盖物,其中含有少量细小的钙颗粒.花粉落到柱头上后,储存在花粉外壁中的钙被释放到覆盖层中.当花粉管穿过覆盖层长入柱头细胞之间时,花粉管顶端的细胞壁中出现了大量的细小钙颗粒.开花后22 h观察时,在花柱引导组织中形成了钙的梯度分布:花柱上部引导组织中的钙较少,而下部连接子房处的花柱引导组织中含有较多的钙颗粒.去雄花开花后1 d时,花柱上部引导组织中的钙明显增多;3 d时,连柱头细胞中也出现了较多的钙颗粒.讨论了烟草花柱引导组织中钙梯度分布和花粉管生长的关系.