Phosphoglucomutase and trehalase isoenzymes of Venezuelan simulium vectors of Onchocerca volvulus

Phosphoglucomutase (PGM) and trehalase (Tre) isoenzymes of five species of Simulium blackflies (Diptera: Simuliidae), vectors of onchocerciasis in Venezuela, were investigated by means of a portable electrophoresis field kit. Tre differed between S. incrustatum and S. oyapockense s.l. Electrophoretic variation of Tre in other members of the S. amazonicum and S. incrustatum groups merit further investigation. PGM appears to be more useful for separating populations within species complexes. Multiple populations and/or seasonal changes in population structure of S. guianense s.l., S. exiguum s.l. and S. metallicum s.l. were inferred from elecrophoretic variation of PGM.     

Further evidence that deltamethrin-impregnated collars protect domestic dogs from sandfly bites

In many foci of zoonotic visceral leishmaniasis (ZVL), domestic dogs are important reservoir hosts of the causative Leishmania parasites transmitted by phlebotomine sandflies (Diptera: Psychodidae). We tested the protective value of impregnated dog collars (20 g plastic containing deltamethrin 800 mg ai) against Phlebotomus papatasi (Scopoli) sandflies in Iran. For each assay, the dog was sedated and caged in a net with 70-100 wild-caught sandflies overnight (23.30-06.30 hours). Dogs wearing the collars were bitten by approximately 80% fewer sandflies than before collars were fitted, i.e. 51% vs. 11% of hungry female flies exposed. Sandfly mortality rates following 20 h exposure to dogs with collars (18%) or without collars (17%) were not significantly different. Effects of collars were tested when dogs had been wearing them for 8 days. A previous trial against the sandfly P. perniciosus Newstead in France, using smaller dogs, showed that effects of such collars were not fully realized until they had been worn for 2 weeks or more; they remained effective for at least 8 months and killed significant proportions of the sandflies exposed. Present results with P. papatasi, confirming that this simple device provides effective protection against sandflies, are considered sufficiently encouraging to justify a community-wide field trial of deltamethrin-impregnated dog collars against ZVL vector sandflies in Iran.     

Diversion of Anopheles gambiae from children to other hosts following exposure to permethrin-treated bednets

Permethrin-treated bednets reduce mortality and morbidity from malaria in Gambian children. However, it is not certain how this effect is achieved, as neither mosquito numbers nor the human blood index of indoor-resting female Anopheles gambiae Giles sensu lato (Diptera: Culicidae) mosquitoes have been reduced when treated bednets were introduced into a community. One possibility is that insecticide-treated bednets divert mosquitoes from children to adults. To investigate this hypothesis, a cross-over trial with insecticide-treated bednets was undertaken in two small Gambian villages. To differentiate mosquitoes that had fed on children from those that had fed on adults, all children in the study villages were immunized with rabies vaccine before the trial. Using the detection of rabies antibody in a bloodmeal as an indicator that a mosquito had bitten a child, it was found that the percentage of blood-fed mosquitoes caught indoors that had bitten a child fell significantly from 30.8% to 9.2% and from 28.0% to 6.9% in each village after insecticide-treated bednets were introduced. To investigate the possibility that some diversion to animals had occurred, a PCR analysis for human beta-globin DNA was undertaken on selected samples. The results of this investigation were confusing, as some rabies-antibody positive bloodmeals were negative for human DNA. This may have been due to cross-reacting antibodies in animal sera and/or DNA degradation by digestion in the mosquito. Although good evidence for diversion of mosquitoes away from children was obtained, it remains uncertain whether diversion was mainly to adult humans, to animals or to both.     

Seasonal Population Dynamics of Ixodes Ticks and Tick-Borne Encephalitis Virus

Seasonality of the epidemic and epizootic processes of tick-borne encephalitis (TBE) depend on the period of activity of ixodid ticks Ixodes persulcatus Schulze and I. ricinus Linnaeus, which are the main reservoirs and vectors of TBE virus, and also on the process of their activation. The period of activity is the period during which the ticks occur in the active state. Activation is the transition into this state of ticks that moulted from the preceding stage and completed post-moulting development. For I. persulcatus, the first adult ticks generally emerge between April 10 and May 9. Under a variety of natural conditions, activation of adult I. persulcatus after wintering lasts for 45–86 days and this period may be even longer in certain areas of the Far East. The period during which one-half of the entire tick population becomes activated (AT₅₀) comprises no more than 10–20 days. In adult I. ricinus ticks the activation period may last even longer than in I. persulcatus. The data on duration of the period of activity and on activation of larval and nymphal stages of both tick species were considered. Ticks exhausting their nutrient reserves and failing to find a host die quickly. The period during which 50% of the entire tick population die under natural conditions is designated LT₅₀. The main types of I. persulcatus and I. ricinus seasonal activity within their species ranges were reviewed. Data on the relationship between TBE virus reproduction in a natural focus and physiological age, pattern of activation, and seasonal changes in age structure of the tick population were analyzed. Seasonal changes in the prevalence of infection among active unfed adult ticks in a natural population are determined by virus content in individual ticks at the moment of their activation and also by the duration of subsequent virus persistence (the rate of virus loss) in ticks. Apparently, the opportunity and frequency of horizontal TBE virus transmission under natural conditions, change during the season of tick activity.     

腺病毒介导的人巨细胞病毒UL49基因小鼠模型的建立

建立表达HCMV UL49 基因的转基因小鼠,为抗病毒药物研究提供有效的实验动物模型。本实验将UL49-GFP基因插入腺病毒穿梭质粒pDC316中,构建重组质粒pDC316-UL49-GFP,与腺病毒骨架质粒pBHGloxΔE1,3Cre 通过脂质体介导共转染293 细胞,重组产生腺病毒Ad-UL49-GFP, 经PCR和Western Blot鉴定正确后,大量扩增、纯化,制备高滴度重组腺病毒。纯化腺病毒经尾静脉注射感染小鼠,通过荧光定量PCR 和Western blot 方法,检测UL49 基因在小鼠体内组织分布和表达时相。结果显示UL49基因在小鼠的心、肝、脾、肺、肾组织均有表达,并且表达量由高到低顺序依次是：肝、脾、肾、心、肺,在腺病毒感染第3天在各靶器官表达水平较高,此后逐渐下降,第14天时仅存在肝和脾中。表明表达UL49基因的小鼠模型构建成功。小鼠模型的成功建立为下一步筛选以UL49基因为靶的抗病毒药物奠定了基础。     

The Biology of the Tospoviruses

The tospoviruses are a diverse, cosmopolitan and economically important genus of plant viruses. In the recent past, interest in the tospoviruses has been rekindled with the resurgence and expansion of tomato spotted wilt virus and the appearance of new tospoviruses, including impatiens necrotic spot. This renewed interest in the tospoviruses, accompanied with the many recent advances in plant virology techniques, particularly those utilising molecular biology, have resulted in a rapid growth of our understanding of these viruses. This paper provides a review of the tospoviruses, encompassing all the major aspects of their biology, including the recent changes in the classification of the genus and current knowledge on molecular biology, vector relations, control and diagnosis.     

Ovitrap surveys of dengue vector mosquitoes in Chiang Mai, northern Thailand: seasonal shifts in relative abundance of Aedes albopictus and Ae. aegypti

Aedes aegypti (L.) and Aedes albopictus (Skuse) were surveyed using ovitraps in residential areas in Chiang Mai, northern Thailand. Egg populations (both species inclusive) remained low in the dry season, but increased/decreased exponentially during the first/latter half of the rainy season, respectively. This seasonal pattern was similar to the seasonal distribution of dengue haemorrhagic fever cases in the area. During the dry season (November-March) Ae.aegypti was dominant in urban and indoor ovitraps. With onset of the rainy season in April, relative abundance of Ae.albopictus increased in rural and outdoor ovitraps. Ae.albopictus displaced Ae.aegypti in the latter half of the rainy season in the rural area. Possible mechanisms to account for this seasonal decline of Ae.aegypti and reciprocal fluctuations in relative abundance of Ae.albopictus are discussed in relation to food availability for larvae in container habitats.     

对分子置换法中积分半径选取方法的探讨

通过对晶胞中帕特逊向量的统计分布的研究,从理论上阐述了根据模型结构单元的线度确定旋转函数的积分半径这种作法的合理性,并且指出了估算积分半径取值范围的具体方法,以及自身旋转函数与交叉旋转函数的积分半径取值范围的区别。经过我们将此方法应用于酚胰岛素Ｂ链羰端六肽胰岛素的旋转函数求解,计算结果证实了这种积分半径的估算方法的可靠性。     

The EF‐1α promoter maintains high‐level transgene expression from episomal vectors in transfected CHO‐K1 cells

In our previous study, we demonstrated that episomal vectors based on the characteristic sequence of matrix attachment regions (MARs) and containing the cytomegalovirus (CMV) promoter allow transgenes to be maintained episomally in Chinese hamster ovary (CHO) cells. However, the transgene expression was unstable and the number of copies was low. In this study, we focused on enhancers, various promoters and promoter variants that could improve the transgene expression stability, expression magnitude (level) and the copy number of a MAR‐based episomal vector in CHO‐K1 cells. In comparison with the CMV promoter, the eukaryotic translation elongation factor 1 α (EF‐1α, gene symbol EEF1A1) promoter increased the transfection efficiency, the transgene expression, the proportion of expression‐positive clones and the copy number of the episomal vector in long‐term culture. By contrast, no significant positive effects were observed with an enhancer, CMV promoter variants or CAG promoter in the episomal vector in long‐term culture. Moreover, the high‐expression clones harbouring the EF‐1α promoter tended to be more stable in long‐term culture, even in the absence of selection pressure. According to these findings, we concluded that the EF‐1α promoter is a potent regulatory sequence for episomal vectors because it maintains high transgene expression, transgene stability and copy number. These results provide valuable information on improvement of transgene stability and the copy number of episomal vectors.     

Efficient adenoviral-mediated gene delivery into porcine mesenchymal stem cells

Mesenchymal stem cell (MSC) mediated gene therapy research has been conducted predominantly on rodents. Appropriate large animal models may provide additional safety and efficacy information prior to human clinical trials. The objectives of this study were: (a) to optimize adenoviral transduction efficiency of porcine bone marrow MSCs using a commercial polyamine-based transfection reagent (GeneJammer, Stratagene, La Jolla, CA), and (b) to determine whether transduced MSCs retain the ability to differentiate into mesodermal lineages. Porcine MSCs (pMSCs) were infected under varying conditions, with replication-defective adenoviral vectors carrying the GFP gene and GFP expression analyzed. Transduced cells were induced to differentiate in vitro into adipogenic, chondrogenic, and osteogenic lineages. We observed a 5.5-fold increase in the percentage of GFP-expressing pMSCs when adenovirus type 5 carrying the adenovirus type 35 fiber (Ad5F35eGFP) was used in conjunction with GeneJammer. Transduction of pMSCs at 10.3-13.8 MOI (1,500-2,000 vp/cell) in the presence of Gene Jammer yielded the highest percentage of GFP-expressing cells ( approximately 90%) without affecting cell viability. A similar positive effect was detected when pMSCs were infected with an Ad5eGFP vector. Presence of fetal bovine serum (FBS) during adenoviral transduction enhanced vector-encoded transgene expression in both GeneJammer-treated and control groups. pMSCs transduced with adenovirus vector in the presence of GeneJammer underwent lipogenic, chondrogenic, and osteogenic differentiation. Addition of GeneJammer during adenoviral infection of pMSCs can revert the poor transduction efficiency of pMSCs while retaining their pluripotent differentiation capacity. GeneJammer-enhanced transduction will facilitate the use of adenoviral vectors in MSC-mediated gene therapy models and therapies.