BACKGROUND & AIMS: -2

背景与目标： -2

BACKGROUND & AIMS: :Somatic gene transfer continues to have potential for the study and therapy of cardiovascular disease. We have developed a modular, self-assembling, nonviral system consisting of Lipofectin, integrin-targeting peptides, and plasmid DNA (LID) and we have applied this to a model of vascular injury, rat carotid angioplasty. Marker gene studies identified transfection of adventitial cells after vector delivery to that layer. Human tissue inhibitor of metalloproteinase-1 (hTIMP-1) was tested as a therapeutic gene product after direct application to the exposed adventitial layer. Vascular LID.hTIMP-1 transfection was confirmed by polymerase chain reaction and gene expression by immunohistochemistry at 7 days. Neointimal areas were 0.160 +/- 0.078 and 0.225 +/- 0.052 mm(2) for LID.hTIMP-1-transfected (n = 14) and LID.pCI-transfected (n = 12) vessels, respectively, at 14 days, and 0.116 +/- 0.068 mm(2) (n = 14) and 0.194 +/- 0.095 mm(2) (n = 14), respectively, at 28 days, representing a 29 and 40% reduction in neointimal hyperplasia at 14 and 28 days, respectively, after balloon dilatation. Neointima-to-media ratios were similarly reduced. In addition, expansile remodeling after balloon injury was inhibited at 14 days, the area within the external elastic lamina being 0.50 +/- 0.02 and 0.61 +/- 0.02 mm(2) in LID.hTIMP-1- and LID.pCI-transfected arteries, respectively (p < 0.0005). We have demonstrated an effective system of therapeutic gene transfer, particularly targeting the arterial adventitia, where transfer of genes involved in matrix remodeling and cell migration may be useful.

背景与目标： ：体细胞基因转移对于心血管疾病的研究和治疗仍然具有潜力。我们已经开发了由脂转染蛋白，整合素靶向肽和质粒DNA（LID）组成的模块化，自组装非病毒系统，并将其应用于血管损伤，大鼠颈动脉血管成形术的模型。标记基因研究确定了载体递送至该层后外膜细胞的转染。直接应用于暴露的外膜层后，将人类组织金属蛋白酶-1（hTIMP-1）抑制剂作为治疗性基因产物进行了测试。在第7天通过聚合酶链反应和基因表达通过免疫组织化学证实了血管LID.hTIMP-1转染。 LID.hTIMP-1转染（n = 14）和LID.pCI转染（n = 12）的血管分别在14天和0.116处的新内膜面积分别为0.160 /-0.078和0.225 /-0.052 mm（2）。 /-在28天时分别为0.068 mm（2）（n = 14）和0.194 /-0.095 mm（2）（n = 14），分别表示在14天和28天时内膜增生减少了29％和40％ ，球囊扩张后。新内膜与培养基的比例也降低了。此外，气囊损伤后的重塑在第14天被抑制，LID.hTIMP-1-和LID.pCI转染的动脉中，外部弹性层的面积分别为0.50 /-0.02和0.61 /-0.02 mm（2），分别为（p <0.0005）。我们已经证明了一种有效的治疗性基因转移系统，尤其是靶向动脉外膜，其中涉及基质重塑和细胞迁移的基因转移可能是有用的。

BACKGROUND & AIMS: BACKGROUND:Deltamethrin 62.5 polymer-enhanced suspension concentrate (SC-PE) is one of the World Health Organization-approved insecticides for indoor residual spraying and was recommended to evaluate its residual activity for determination of appropriate spray cycles in different eco-epidemiologic settings. In the current study, efficacy of deltamethrin 62.5 SC-PE was evaluated against vectors of malaria and its impact on malaria incidence in a Plasmodium falciparum hyper-endemic area in Koraput district, Odisha State, India. METHODS:The trial had two comparable arms, arm 1 with residual spraying of deltamethrin 62.5 SC-PE and arm 2 with deltamethrin 2.5% WP (positive control). Comparative assessment of the impact of each intervention arm on entomological (density, parity, infection and human blood index), epidemiological (malaria incidence) parameters, residual efficacy and adverse effects were evaluated. RESULTS:Both the arms were comparable in terms of entomological and epidemiological parameters. While, deltamethrin 62.5 SC-PE was found to be effective for 150 days in mud and wood surfaces and 157 days in cement surfaces; deltamethrin 2.5% was effective only for 105 days on mud surfaces and 113 days on cement and wood surfaces. CONCLUSIONS:Deltamethrin 62.5 SC-PE had prolonged killing effectiveness up to 5 months. Hence, one round of IRS with deltamethrin 62.5 SC-PE would be sufficient to cover two existing malaria peak transmission seasons (July-August and October-November) in many parts of India.

背景与目标： 背景：溴氰菊酯62.5聚合物增强悬浮剂（SC-PE）是世界卫生组织批准的室内残留喷洒杀虫剂之一，建议评估其残留活性，以确定在不同的生态流行病学环境中适当的喷洒周期。在当前的研究中，在印度奥里萨邦科拉普特地区的恶性疟原虫高流行区，对溴氰菊酯62.5 SC-PE的抗疟疾载体及其对疟疾发病率的影响进行了评估。
方法：该试验有两个可比的手臂，手臂1残留喷洒了溴氰菊酯62.5 SC-PE，手臂2喷洒了溴氰菊酯2.5％WP（阳性对照）。比较评估每个干预措施对昆虫学（密度，均等，感染和人类血液指数），流行病学（疟疾发病率）参数，残留功效和不良影响的影响。
结果：在昆虫学和流行病学参数方面，两臂具有可比性。而溴氰菊酯62.5 SC-PE被发现在泥浆和木材表面150天有效，在水泥表面157天有效； 2.5％溴氰菊酯仅在泥浆表面上有效105天，在水泥和木材表面上有效113天。
结论：溴氰菊酯62.5 SC-PE的杀灭效果已延长至5个月。因此，一轮带有溴氰菊酯62.5 SC-PE的IRS足以覆盖印度许多地方的两个现有疟疾高峰传播季节（7月至8月和10月至11月）。

BACKGROUND & AIMS: :A mixed phase hybridization technique was developed to detect dengue virus type 2 (DEN-2) RNA in pools of infected Aedes albopictus mosquitoes using radiolabelled RNA probes. This technique used a guanidine thiocyanate extraction procedure to simplify analyte preparation. The probes contained sequences complementary to portions of the NS-1 or NS-5 genes of the DEN-2 viral genome. One infected mosquito in a pool of 25 could be detected in approximately 48 h. RNAs from DEN serotypes 1-4 were extracted from cultured mosquito (C6/36) cells. The NS-1 RNA probe was highly specific for DEN-2 RNA. The NS-5 RNA probe detected both DEN-2 and DEN-4 RNA. DEN-2 RNA was also detected by molecular hybridization in concentrated solutions of guanidine thiocyanate using the NS-1 probe. Solution hybridization was 10-fold more sensitive when detecting RNA from purified DEN-2 virus than the mixed phase assay and could detect one infected mosquito in a pool of 25 within 6-8 h. Solution hybridizations were performed in 2-3 h vs 16-20 h for mixed phase hybridizations, and solution hybridizations required 5-10 times less mosquito RNA than mixed phase hybridizations to attain comparable sensitivities. However, solution hybridizations did result in a broader probe specificity than mixed phase hybridizations.

背景与目标： ：开发了一种混合相杂交技术，以使用放射性标记的RNA探针在感染的白纹伊蚊的池中检测登革热病毒2型（DEN-2）RNA。该技术使用了硫氰酸胍提取程序来简化分析物的制备。探针包含与DEN-2病毒基因组的NS-1或NS-5基因的部分互补的序列。在大约48小时内，可以在25个池中检测到一只被感染的蚊子。从培养的蚊子（C6 / 36）细胞中提取DEN血清型1-4的RNA。 NS-1 RNA探针对DEN-2 RNA具有高度特异性。 NS-5 RNA探针同时检测到DEN-2和DEN-4 RNA。还使用NS-1探针通过在硫氰酸胍浓缩溶液中进行分子杂交来检测DEN-2 RNA。当从纯化的DEN-2病毒中检测RNA时，溶液杂交的灵敏度比混合相测定高10倍，并且可以在6-8小时内在25个池中检测到一只被感染的蚊子。溶液杂交在2-3小时内进行，而混合相杂交在16至20小时内进行，与杂交相杂交相比，溶液杂交所需的蚊子RNA少5-10倍，以达到可比的敏感性。但是，溶液杂交的确比混合相杂交产生了更广泛的探针特异性。

BACKGROUND & AIMS: :Neutralizing antibodies directed against adeno-associated virus (AAV) are commonly found in humans. In seropositive subjects, vector administration is not feasible as antibodies neutralize AAV vectors even at low titers. Consequently, a relatively large proportion of humans is excluded from enrollment in clinical trials and, similarly, vector redosing is not feasible because of development of high-titer antibodies following AAV vector administration. Plasmapheresis has been proposed as strategy to remove anti-AAV antibodies from the bloodstream. Although safe and relatively effective, the technology has some limitations mainly related to the nonspecific removal of all circulating IgG. Here we developed an AAV-specific plasmapheresis column which was shown to efficiently and selectively deplete anti-AAV antibodies without depleting the total immunoglobulin pool from plasma. We showed the nearly complete removal of anti-AAV antibodies from high titer purified human IgG pools and plasma samples, decreasing titers to levels that allow AAV vector administration in mice. These results provide proof-of-concept of a method for the AAV-specific depletion of neutralizing antibodies in the setting of in vivo gene transfer.

背景与目标： ：针对腺相关病毒（AAV）的中和抗体通常在人类中发现。在血清反应阳性的受试者中，由于抗体即使在低滴度下也能中和AAV载体，因此载体给药是不可行的。因此，相对较大比例的人被排除在临床试验之外，并且类似地，由于施用AAV载体后高滴度抗体的发展，载体的重新分配是不可行的。血浆置换已被提议作为从血流中去除抗AAV抗体的策略。尽管安全且相对有效，但该技术仍存在一些局限性，主要与所有循环IgG的非特异性清除有关。在这里，我们开发了一种AAV特异性血浆置换色谱柱，该色谱柱可有效且选择性地消耗抗AAV抗体，而不会消耗血浆中的总免疫球蛋白。我们显示了从高滴度纯化的人IgG库和血浆样品中几乎完全去除了抗AAV抗体，将滴度降低到允许在小鼠中施用AAV载体的水平。这些结果提供了在体内基因转移中AAV特异性消耗中和抗体的方法的概念证明。

BACKGROUND & AIMS: :Incorporation of a central polypurine tract (cPPT) and a posttranscriptional regulatory element (PRE) into lentivirus vectors provides increased transduction efficiency and transgene expression. We compared the effects of these elements individually and together on transduction efficiency and gene expression, using lentivirus vectors pseudotyped with vesicular stomatitis virus G protein (VSV-G) and encoding enhanced green fluorescent protein (GFP) and rat erythropoietin (EPO). The transduction efficiency was greater than 2-fold higher in the vector containing the PRE element, 3-fold higher in vector encoding the cPPT element, and 5-fold increased in the GFP virus containing both cPPT and PRE elements relative to the parent virus. In comparison with parent vector the mean fluorescence intensity (MFI) of GFP expression was 7-fold higher in cells transduced with virus containing PRE, 6-fold increased in cells transduced with virus containing cPPT, and 42-fold increased in GFP-virus containing both cPPT and PRE elements. EPO-virus containing a PRE element showed a nearly 5-fold increase in EPO secretion over the parent vector, and the vector encoding both PRE and cPPT showed a 65-fold increase. Thus, lentivirus vectors incorporating both PRE and cPPT showed expression levels significantly increased over the sum of the components alone, suggesting a synergistic effect.

背景与目标： 将慢病毒载体整合到中央多嘌呤区（cPPT）和转录后调控元件（PRE）中，可提高转导效率和转基因表达。我们使用伪装了水泡性口炎病毒G蛋白（VSV-G）并编码增强型绿色荧光蛋白（GFP）和大鼠促红细胞生成素（EPO）的慢病毒载体，比较了这些元素单独以及一起对转导效率和基因表达的影响。相对于亲本病毒，在含有PRE元素的载体中，转导效率高出2倍，在编码cPPT元素的载体中，转导效率高出3倍，在同时含有cPPT和PRE元素的GFP病毒中，转导效率提高了5倍。与亲代载体相比，用含PRE的病毒转导的细胞中GFP表达的平均荧光强度（MFI）高7倍，用含cPPT的病毒转导的细胞中GFP表达的平均荧光强度高6倍，而在含cPPT的病毒转导的细胞中GFP表达的平均荧光强度增加42倍。 cPPT和PRE元素。含有PRE元件的EPO病毒的EPO分泌量比亲本载体增加了近5倍，而编码PRE和cPPT的载体均表现出了65倍的增加。因此，掺有PRE和cPPT的慢病毒载体显示表达水平明显高于单独组分的总和，表明具有协同作用。

BACKGROUND & AIMS: :Pathologic lesions caused by catheter-based revascularization procedures for occlusive artery disease include disruption of the endothelium, exposure of extracellular matrix (ECM) proteins, and proliferation of vascular smooth muscle cells, which lead to neointima formation and restenosis. We have developed matrix-collagen-targeted retroviral vectors that are able to accumulate at sites of vascular injury (Hall et al., Hum. Gene Ther. 1997;8:2183-2192; Hall et al., Hum. Gene Ther. 2000;11:983-993). The primary tissue-targeting motif, adapted from the physiological surveillance sequence found in von Willebrand factor, served to localize and concentrate the vector within vascular lesions. In the present study, we evaluated the efficiency of this vector-targeting system in rats with nonligated balloon-injured carotid arteries. Both intraarterial (by retrograde femoral artery catheterization) and intravenous (via femoral vein) injection of a matrix-targeted vector enhanced transduction of neointimal cells ( approximately 20%) at severely denuded areas when compared with the nontargeted vector (<1%). Further, intraarterial instillation of a matrix-targeted, but not a nontargeted, vector bearing an antisense cyclin G1 construct inhibited neointima lesion formation in the injured carotid arteries. Taken together, these data indicate that strategic targeting of retroviral vectors to vascular lesions would have therapeutic potential in the management of vascular restenosis and many other disorders of uncontrolled proliferation where endothelial disruption, ECM remodeling, and collagen deposition form the nexus for preferential vector localization and concentration in vivo.

背景与目标： 闭塞性动脉疾病的基于导管的血运重建术引起的病理损害包括内皮破坏，细胞外基质（ECM）蛋白暴露以及血管平滑肌细胞增殖，从而导致新内膜形成和再狭窄。我们已经开发了靶向胶原蛋白的逆转录病毒载体，它们能够在血管损伤部位积聚（Hall等，Hum。Gene Ther。1997; 8：2183-2192； Hall等，Hum.Gene Ther。2000 ; 11：983-993）。适应于von Willebrand因子中发现的生理学监视序列的主要组织靶向基序可将载体定位并集中在血管病变内。在本研究中，我们评估了这种载体靶向系统在未结扎球囊损伤颈动脉的大鼠中的效率。与非靶向载体（<1％）相比，动脉内注射（通过逆行股动脉导管插入术）和静脉内（通过股静脉）注射在严重裸露区域的新内膜细胞（约20％）的转导都增强了内膜细胞的转导。此外，动脉内滴注带有反义细胞周期蛋白G1构建体的靶向基质的载体，而非非靶向载体，抑制了受损颈动脉中新内膜病变的形成。综上所述，这些数据表明，将逆转录病毒载体战略性地靶向血管病变将具有治疗血管再狭窄和许多其他不受控制的增生性疾病的治疗潜力，其中内皮细胞破坏，ECM重塑和胶原蛋白沉积形成了优先载体定位和相关性的纽带。体内浓度。

BACKGROUND & AIMS: :In this study we analyzed two ways of retargeting of Ad-vectors to human pancreatic carcinoma with the aim of enhancing the gene transfer efficiency. First, we analyzed the expression of the epidermal growth factor receptor (EGFR) on primary, as well as established pancreatic carcinoma cells by flow cytometry which revealed high expression levels of EGFR on the surface of these cells. We showed that EGFR-retargeted entry pathway using a bispecific fusion protein formed by a recombinant soluble form of truncated Coxsackie and Adenovirus Receptor (sCAR) genetically fused with human EGF (sCAR-EGF) redirects them to the EGFR leading to an enhanced gene transfer efficiency to pancreatic carcinoma cells. Since flow cytometry revealed absence of CAR expression, but the presence of at least one of both alphav integrins on the pancreatic carcinoma cells, a second way of targeting was investigated using a genetically modified Ad vector which has an RGD (Arg-Gly-Asp)-containing peptide inserted into the HI-loop of the fiber knob. This RGD targeted Ad (AdlucRGD) revealed efficient CAR-independent infection by allowing binding to cellular integrins resulting in a dramatic enhancement of gene transfer. These findings have direct relevance for Ad-vector based gene therapy strategies for pancreatic carcinoma.

背景与目标： ：在这项研究中，我们分析了将Ad载体重新靶向人胰腺癌的两种方法，目的是提高基因转移效率。首先，我们通过流式细胞术分析了表皮生长因子受体（EGFR）在原发性和已建立的胰腺癌细胞上的表达，揭示了这些细胞表面上EGFR的高表达水平。我们显示，使用由人源EGF（sCAR-EGF）基因融合的截短的柯萨奇和腺病毒受体（sCAR）的重组可溶性形式形成的双特异性融合蛋白，通过双特异性融合蛋白形成的EGFR靶向进入途径将其重定向至EGFR，从而提高了基因转移效率胰腺癌细胞。由于流式细胞仪显示不存在CAR表达，但在胰腺癌细胞上同时存在两种alphav整合素中的至少一种，因此使用具有RGD（Arg-Gly-Asp）的基因修饰的Ad载体研究了第二种靶向方法插入到纤维结的HI-环中的含-肽。该RGD靶向广告（AdlucRGD）通过与细胞整联蛋白结合，从而显着增强了基因转移，从而揭示了与CAR无关的有效感染。这些发现与基于Ad载体的胰腺癌基因治疗策略直接相关。

BACKGROUND & AIMS: BACKGROUND:Knockdown resistance (kdr) is a well-characterized target-site insecticide resistance mechanism that is associated with DDT and pyrethroid resistance. Even though insecticide resistance to pyrethroids and DDT have been reported in Anopheles albimanus, Anopheles benarrochi sensu lato (s.l.), Anopheles darlingi, Anopheles nuneztovari s.l., and Anopheles pseudopunctipennis s.l. malaria vectors in Latin America, there is a knowledge gap on the role that kdr resistance mechanisms play in this resistance. The aim of this study was to establish the role that kdr mechanisms play in pyrethroid and DDT resistance in the main malaria vectors in Colombia, in addition to previously reported metabolic resistance mechanisms, such as mixed function oxidases (MFO) and nonspecific esterases (NSE) enzyme families. METHODS:Surviving (n = 62) and dead (n = 67) An. nuneztovari s.l., An. darlingi and An. albimanus mosquitoes exposed to diagnostic concentrations of DDT and pyrethroid insecticides were used to amplify and sequence a ~ 225 bp fragment of the voltage-gated sodium channels (VGSC) gene. This fragment spanning codons 1010, 1013 and 1014 at the S6 segment of domain II to identify point mutations, which have been associated with insecticide resistance in different species of Anopheles malaria vectors. RESULTS:No kdr mutations were detected in the coding sequence of this fragment in 129 samples, 62 surviving mosquitoes and 67 dead mosquitoes, of An. darlingi, An. nuneztovari s.l. and An. albimanus. CONCLUSION:Mutations in the VGSC gene, most frequently reported in other species of the genus Anopheles resistant to pyrethroid and DDT, are not associated with the low-intensity resistance detected to these insecticides in some populations of the main malaria vectors in Colombia. These results suggest that metabolic resistance mechanisms previously reported in these populations might be responsible for the resistance observed.

背景与目标： 背景：击倒抗性（kdrdown）是一种特征明确的靶点杀虫剂抗性机制，与DDT和拟除虫菊酯抗性相关。尽管在白bi按蚊，按蚊Bennarrochi sensu lato（s.l.），达令按蚊Annueles narlingtovari s.l.和拟按蚊Anopheles pseudopunctipennis s.l.中已经报告了对拟除虫菊酯和DDT的杀虫剂抗性。在拉丁美洲的疟疾媒介中，关于kdr抗性机制在这种抗性中所起作用的知识差距很大。这项研究的目的是确定哥伦比亚除主要报道的代谢抗性机制（如混合功能氧化酶（MFO）和非特异性酯酶（NSE））外，kdr机制在哥伦比亚主要疟疾媒介中对拟除虫菊酯和滴滴涕抗性中的作用。酶家族。
方法：存活（n = 62）和死亡（n = 67）。 nuneztovari s.l.，An。达令吉和安。暴露于诊断浓度的滴滴涕和拟除虫菊酯杀虫剂的白bi蚊被用来扩增和测序电压门控钠通道（VGSC）基因的〜225 bp片段。该片段跨越域II的S6区段的密码子1010、1013和1014，以识别点突变，该突变已与不同种类的按蚊疟疾载体中的杀虫剂抗性相关。
结果：在An的129个样品，62个存活的蚊子和67个死亡的蚊子中，在该片段的编码序列中未检测到kdr突变。达令吉努涅斯托瓦里湖和。 albimanus。
结论：VGSC基因的突变（在拟除虫菊酯和滴滴涕耐药的其他按蚊属的其他物种中最常报道）与在哥伦比亚一些主要疟疾媒介人群中对这些杀虫剂的低强度耐药性无关。这些结果表明，先前在这些人群中报道的代谢抗性机制可能是观察到的抗性的原因。

BACKGROUND & AIMS: :In the past decade, adenovirus vectors have generated tremendous interest, especially in gene therapy applications. In the so-called 'first generation' adenovirus vectors, the transgenes are inserted in place of the E1 region, or less often the E3 region. Although second-generation and helper-dependent adenovirus vectors will probably prevail in the future in applications that require long-term gene expression, first generation adenovirus vectors will remain very useful in other settings, such as cancer and vaccination, or simply to transfect cell lines that are refractory to other transfection methods. Until a few years ago, the construction of first generation adenovirus vectors was a labor-intensive and time-consuming process. More than 20 methods have appeared that facilitate their construction and are reviewed below.

背景与目标： 在过去的十年中，腺病毒载体引起了极大的兴趣，特别是在基因治疗应用中。在所谓的“第一代”腺病毒载体中，转基因被插入到E1区域，或更不常见的是E3区域。尽管第二代和依赖于助手的腺病毒载体可能会在将来需要长期表达基因的应用中盛行，但第一代腺病毒载体在其他环境（例如癌症和疫苗接种）或仅用于转染细胞系中仍将非常有用其他转染方法无法耐受的直到几年前，第一代腺病毒载体的构建还是一项劳动密集型且耗时的过程。已经出现了20多种有助于其构建的方法，下面将进行综述。

BACKGROUND & AIMS: :Currently, mosquito vector control is facing a number of key challenges, including the rapid development of resistance to synthetic pesticides and the recent spread of aggressive arbovirus outbreaks. The biosynthesis of silver nanoparticles (AgNPs) is currently considered an environmental friendly alternative to the employ of pyrethroids, carbamates and microbial agents (e.g. Bacillus thuringiensis var. israelensis), since AgNPs are easy to produce, effective and stable in the aquatic environment. However, their biophysical features showed wide variations according to the botanical agent using for the green synthesis, outlining the importance of screening local floral resources used as reducing and stabilizing agents. In this study, we focused on the biophysical properties and the mosquitocidal action of Quisqualis indica-fabricated AgNPs. AgNPs were characterized using spectroscopic (UV, FTIR, XRD) and microscopic (AFM, SEM, TEM and EDX) techniques. AFM, SEM and TEM confirmed the synthesis of poly-dispersed AgNPs with spherical shape and size ranging from 1 to 30nm. XRD shed light on the crystalline structure of these AgNPs. The acute toxicity of Quisqualis indica extract and AgNPs was evaluated against malaria, arbovirus, and filariasis vectors, Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus, as well as on three important non-target aquatic organisms. The Q. indica leaf extract showed moderate larvicidal effectiveness on Cx. quinquefasciatus (LC50=220.42), Ae. aegypti (LC50=203.63) and An. stephensi (LC50=185.98). Q. indica-fabricated AgNPs showed high toxicity against Cx. quinquefasciatus (LC50=14.63), Ae. aegypti (LC50=13.55) and An. stephensi (LC50=12.52), respectively. Notably, Q. indica-synthesized AgNPs were moderately toxic to non-target aquatic mosquito predators Anisops bouvieri (LC50=653.05μg/mL), Diplonychus indicus (LC50=860.94μg/mL) and Gambusia affinis (LC50=2183.16μg/mL), if compared to the targeted mosquitoes. Overall, the proposed one-pot biogenic fabrication of AgNPs using Q. indica is a low-cost and eco-friendly tool in the fight against Zika virus, malaria and filariasis vectors, with little impact against non-target aquatic mosquito predators.

背景与目标： 目前，蚊媒的控制面临许多关键挑战，包括对合成农药的抗性迅速发展以及近期侵略性虫媒病毒爆发的蔓延。银纳米颗粒（AgNPs）的生物合成目前被认为是替代拟除虫菊酯，氨基甲酸酯和微生物制剂（例如苏云金芽孢杆菌var.israelensis）的一种环境友好替代方法，因为AgNPs在水生环境中易于生产，有效且稳定。然而，根据用于绿色合成的植物剂，它们的生物物理特征显示出很大的差异，突出了筛选用作还原剂和稳定剂的当地花卉资源的重要性。在这项研究中，我们集中于Quisqualis d制造的AgNPs的生物物理特性和灭蚊作用。 AgNPs使用光谱技术（UV，FTIR，XRD）和微观技术（AFM，SEM，TEM和EDX）进行表征。 AFM，SEM和TEM证实了球形和尺寸范围为1至30nm的多分散AgNP的合成。 XRD揭示了这些AgNP的晶体结构。评价了Quisqualis indica提取物和AgNPs对疟疾，虫媒病毒和丝虫病载体，Stephensi按蚊，埃及伊蚊和库克西库克斯犬以及三种重要的非目标水生生物的急性毒性。 in叶提取物对Cx表现出中等杀幼虫效果。 quinquefasciatus（LC50 = 220.42），Ae。埃及（LC50 = 203.63）和An。斯蒂芬斯（LC50 = 185.98）。 Q. Ag制造的AgNP对Cx表现出高毒性。 quinquefasciatus（LC50 = 14.63），Ae。埃及（LC50 = 13.55）和An。斯蒂芬斯（LC50 = 12.52）。值得注意的是，Q合成的AgNPs对非目标水生捕食性蚊食美洲大按蚊（LC50 =653.05μg/ mL），印度洋双翅目（LC50 =860.94μg/ mL）和革兰（Gambusia affinis）（LC50 =2183.16μg/ mL）具有中等毒性。 ，如果与目标蚊子相比。总体而言，拟议的使用印度Q草的一锅式AgNP的生物制备是对抗寨卡病毒，疟疾和丝虫病载体的一种低成本且生态友好的工具，对非目标水生蚊虫几乎没有影响。

BACKGROUND & AIMS: :RNA editing is a source of molecular diversity that regulates the functional repertoire of animal transcriptomes. Multiple studies in Drosophila have revealed that conserved editing events can be a source of evolutionary adaptations, and there is a solid body of evidence linking editing and the fine-tuning of neural genes, which are often targeted by insecticides used in vector control. Yet, despite these suggestive connections, genome-wide analyses of editing in insect vectors are conspicuously lacking. Future advances will require complementing the growing wealth of vector genomes with targeted transcriptome analyses. Here, we review recent investigations of the genetic footprints of adaptive RNA editing in insects and provide an overview of new methodologies applicable to studies of RNA editing in insect vectors.

背景与目标： RNA编辑是调节动物转录组功能的分子多样性的一个来源。在果蝇中的多项研究表明，保守的编辑事件可能是进化适应的来源，并且存在将编辑和神经基因的微调联系起来的可靠证据，而神经基因通常是载体控制中使用的杀虫剂所针对的。然而，尽管有这些暗示性的联系，但是仍然明显缺乏对昆虫载体编辑的全基因组分析。未来的发展将需要通过靶向转录组分析来补充日益丰富的载体基因组。在这里，我们回顾了昆虫中自适应RNA编辑的遗传足迹的最新研究，并提供了适用于昆虫载体RNA编辑研究的新方法的概述。

BACKGROUND & AIMS: :Up until recently, Australia was considered free of Leishmania due to the absence of phlebotomine sandfly species (Diptera: Phlebotominae) known to transmit Leishmania parasites in other parts of the world. The discovery of Leishmania (Mundinia) macropodum (Kinetoplastida: Trypanosomatidae) in Northern Australia sparked questions as to the existence of alternative vectors of Leishmania. This has added to the complexity of fully understanding the parasite's interaction with its vector, which is known to be very specific. Previous findings demonstrated L. macropodum infection beyond the blood meal stage in the day-biting midges Forcipomyia (Lasiohelea) Kieffer (Diptera: Ceratopogonidae) implicating them in the parasite's life cycle. Currently, there is no conclusive evidence demonstrating this suspected vector to transmit L. macropodum to a naïve host. Therefore, this research aimed to investigate the vector competency of day-biting midge F. (Lasiohelea) to transmit L. macropodum utilising a novel technology that preserves nucleic acids. Honey-soaked Flinders Technology Associates (FTA®) filter-paper cards were used to obtain saliva expectorated from biting midges while sugar-feeding. F. (Lasiohelea) were aspirated directly off macropods from a known Leishmania-transmission site and were kept in a waxed-paper container holding a honey-coated FTA® card for feeding. Insect identification and Taqman quantitative real-time PCR (qPCR) screening assays revealed L. macropodum DNA in F. (Lasiohelea) up to 7 days post field-collection, and in an unidentified biting midge, previously known as F. (Lasiohelea) sp.1. Moreover, 7/145 (4.83%) of FTA® cards were confirmed positive with L. macropodum DNA after exposure to field-collected F. (Lasiohelea). Additionally, FTA® cards were found to be a valuable surveillance tool, given the ease of use in the field and laboratory. Overall, our findings support previous reports on L. macropodum transmission by an alternative vector to phlebotomine sandflies. Further studies identifying and isolating infective L. macropodum promastigotes is necessary to resolve questions on the L. macropodum vector.

背景与目标： ：直到最近，由于在世界其他地方没有已知传播利什曼原虫寄生虫的毒理mine沙蝇物种（双翅目：Phlebotominae），澳大利亚被认为没有利什曼原虫。在北澳大利亚发现利什曼原虫（Mundinia）macropodum（Kinetoplastida：Trypanosomatidae）引发了人们对利什曼原虫替代载体的存在的疑问。这增加了充分了解寄生虫与其媒介的相互作用的复杂性，这是众所周知的。先前的发现表明，在咬咬mid虫（Lasiohelea）Kieffer（双翅目：Ceratopogonidae）的食血期之后，L。macropodum感染超过了寄生虫的生命周期。目前，尚无确凿证据表明该可疑载体将大脚乳杆菌传播至幼稚宿主。因此，本研究旨在利用一种保存核酸的新技术来研究咬日mid（Lasiohelea）传播大花胶球菌的载体能力。蜂蜜浸透的Flinders Technology Associates（FTA®）滤纸卡用于在喂糖时从咬咬mid虫中排出唾液。将F.（Lasiohelea）从已知的利什曼原虫传播地点直接从巨足类动物中吸出，并保存在装有蜂蜜涂层FTA®卡的蜡纸容器中进行喂养。昆虫鉴定和Taqman定量实时PCR（qPCR）筛选分析揭示了田间采集后长达7天的F.（Lasiohelea）中的L. macropodum DNA，以及在以前被称为F.（Lasiohelea）sp的不明咬人蚊中。 .1。此外，在暴露于田间采集的F.（Lasiohelea）后，证实了L. macropodum DNA的FTA®卡有7/145（4.83％）阳性。此外，由于FTA®卡在现场和实验室易于使用，因此被认为是一种有价值的监视工具。总的来说，我们的研究结果支持了以前关于巨脚乳杆菌通过除草胺sand替代载体传播的报道。进一步的研究，以鉴定和分离感染性大脚紫苏前鞭毛体对于解决关于大脚紫苏载体的问题是必要的。

BACKGROUND & AIMS: :A new Doppler echocardiographically based method has been developed to quantify volume flow rate by surface integration of velocity vectors (SIVV). Electrocardiographic-gated color Doppler images acquired in two orthogonal planes were used to estimate volume flow rate through a bowl-shaped surface at a given time and distance from the probe. To provide in vitro validation, the method was tested in a hydraulic model representing a pulsatile flow system with a restrictive orifice. Accurate estimates of stroke volume (+/- 10%) were obtained in a window between 1.2 and 1.6 cm proximal to the orifice, just before the region of prestenotic acceleration. By use of the Bernoulli's equation, the estimated flows were used to generate pressure gradient waveforms across the orifice, which agreed well with the measured flows. To demonstrate in vivo applicability, the SIVV method was applied retrospectively to the determination of stroke volume and subaortic flow from the apical three-chamber and five-chamber views in two patients. Stroke volume estimates along the left ventricular outflow tract showed a characteristic similar to that in the in vitro study and agreed well with those obtained by the Fick oxygen method. The region where accurate measurements can be obtained is affected by instrumental factors including Nyquist velocity limit, wall motion filter cutoff, and color flow sector angle. The SIVV principle should be useful for quantitative assessment of the severity of valvular abnormalities and noninvasive measurement of pulsatile volume flows in general.

背景与目标： ：已经开发了一种基于多普勒超声心动图的新方法，可以通过对速度矢量（SIVV）进行表面积分来量化体积流量。在两个正交平面中采集的心电门控彩色多普勒图像用于估计在给定时间和距探头的距离上通过碗状表面的体积流量。为了提供体外验证，该方法在液压模型中进行了测试，该液压模型表示带有节流孔的脉动流系统。在靠近小孔的区域内，在靠近孔口1.2到1.6厘米之间的窗口中获得了对搏动量的准确估计（±10％）。通过使用伯努利方程，估计流量用于生成穿过孔口的压力梯度波形，该波形与测得的流量非常吻合。为了证明在体内的适用性，将SI​​VV方法追溯应用于两名患者从心尖三腔和五腔视图确定中风量和主动脉流量。沿左心室流出道的中风量估计值显示了与体外研究相似的特征，并且与通过Fick氧气法获得的结果相吻合。可以得到准确测量结果的区域受仪器因素的影响，这些因素包括奈奎斯特速度极限，壁运动滤波器截止和色流扇形角。一般而言，SIVV原理可用于定量评估瓣膜异常的严重程度和无创测量搏动量流量。

BACKGROUND & AIMS: :Development of methods to engineer gamma-retroviral vectors capable of transducing target cells in a cell-specific manner could impact the future of the clinical application of gene therapy as well as the understanding of the biology of transfer gene vectors. Two molecular events are critical for controlling the entry of gamma-retroviral vectors to target cells: binding to cell-surface receptors and the subsequent fusion of viral vector membrane and cellular membrane. In this report, we evaluated a method to incorporate a membrane-bound antibody and a fusogenic molecule to provide binding and fusion functions respectively, into gamma-retroviral vectors for targeted gene delivery. An anti-CD20 antibody and a fusogenic protein derived from Sindbis virus glycoprotein could be efficiently co-displayed on the surface of viral vectors. Vectors bearing anti-CD20 antibody conferred their binding specificity to cells expressing CD20. Enhanced in vitro transduction towards CD20-expressing cells was observed for gamma-retroviral vectors displaying both an antibody and a fusogen. We found that the biological activity of the fusogen played an important role on the efficiency of such a targeting strategy and were able to engineer several mutant forms of the fusogen exhibiting elevated fusion function to improve the overall efficiency of targeted transduction. We devised an animal model to show that subcutaneous injection of such engineered vectors to the areas xenografted with target cells could achieve targeted gene delivery in vivo. Taken together, we demonstrated as proof-of-principle a flexible and modular two-molecule strategy for engineering targeting gamma-retroviral vectors.

背景与目标： ：开发能够以细胞特异性方式转导靶细胞的γ-逆转录病毒载体的工程方法，可能会影响基因治疗临床应用的未来以及对转移基因载体生物学的理解。两个分子事件对于控制γ-逆转录病毒载体进入靶细胞至关重要：与细胞表面受体的结合以及随后病毒载体膜和细胞膜的融合。在本报告中，我们评估了将结合膜的抗体和融合分子分别提供结合和融合功能的方法掺入用于目标基因递送的γ-逆转录病毒载体中的方法。抗CD20抗体和Sindbis病毒糖蛋白衍生的融合蛋白可以有效地在病毒载体表面共展示。带有抗CD20抗体的载体赋予它们与表达CD20的细胞的结合特异性。对于展示抗体和融合蛋白的γ-逆转录病毒载体，观察到了向表达CD20的细胞增强的体外转导。我们发现，融合蛋白的生物学活性在这种靶向策略的效率中起着重要作用，并且能够工程化融合蛋白的几种突变形式，这些突变体表现出升高的融合功能，从而提高了靶向转导的整体效率。我们设计了一种动物模型来表明，将这种工程载体皮下注射到异种移植有靶细胞的区域，可以在体内实现靶向基因的递送。两者合计，我们证明了针对工程学针对γ-逆转录病毒载体的灵活模块化的两分子策略作为原理证明。