Fluorescent in situ Hybridization on Mitotic Chromosomes of Mosquitoes

Fluorescent in situ hybridization (FISH) is a technique routinely used by many laboratories to determine the chromosomal position of DNA and RNA probes. One important application of this method is the development of high-quality physical maps useful for improving the genome assemblies for various organisms. The natural banding pattern of polytene and mitotic chromosomes provides guidance for the precise ordering and orientation of the genomic supercontigs. Among the three mosquito genera, namely Anopheles, Aedes, and Culex, a well-established chromosome-based mapping technique has been developed only for Anopheles, whose members possess readable polytene chromosomes ¹. As a result of genome mapping efforts, 88% of the An. gambiae genome has been placed to precise chromosome positions ^2,3 . Two other mosquito genera, Aedes and Culex, have poorly polytenized chromosomes because of significant overrepresentation of transposable elements in their genomes ^{4, 5, 6}. Only 31 and 9% of the genomic supercontings have been assigned without order or orientation to chromosomes of Ae. aegypti ⁷ and Cx. quinquefasciatus ⁸, respectively. Mitotic chromosome preparation for these two species had previously been limited to brain ganglia and cell lines. However, chromosome slides prepared from the brain ganglia of mosquitoes usually contain low numbers of metaphase plates ⁹. Also, although a FISH technique has been developed for mitotic chromosomes from a cell line of Ae. aegypti ¹⁰, the accumulation of multiple chromosomal rearrangements in cell line chromosomes ¹¹ makes them useless for genome mapping. Here we describe a simple, robust technique for obtaining high-quality mitotic chromosome preparations from imaginal discs (IDs) of 4^th instar larvae which can be used for all three genera of mosquitoes. A standard FISH protocol ¹² is optimized for using BAC clones of genomic DNA as a probe on mitotic chromosomes of Ae. aegypti and Cx. quinquefasciatus, and for utilizing an intergenic spacer (IGS) region of ribosomal DNA (rDNA) as a probe on An. gambiae chromosomes. In addition to physical mapping, the developed technique can be applied to population cytogenetics and chromosome taxonomy/systematics of mosquitoes and other insect groups.     

Inhibition of human lymphocyte activation by wheat germ agglutinin: a model for saccharide-specific suppressor factors

The suppressive effect of wheat germ agglutinin (WGA) on lectin-stimulated blastogenesis and immunoglobulin production was studied. Addition of WGA at 10 micrograms/ml inhibited phytohemagglutinin (PHA)-, concanavalin-A (Con-A)-, and pokeweed mitogen (PWM)-induced mitogenic responses by 70-80%. PWM-driven immunoglobulin synthesis was suppressed by 45% with WGA. The inhibitory effects of WGA were not due to cell death or to interference with lectin binding at the cell surface. Inhibition was dependent on the presence of WGA in the cell culture during the first 24 hr of mitogen exposure and was observed in cultures of both monocyte-depleted peripheral blood mononuclear cells as well as T-cell-enriched populations. WGA-induced inhibition of blastogenesis was blocked by the addition of N-acetylglucosamine (GluNAc) which prevents WGA binding to the cell surface. WGA was found to mimic the suppressive effect of a soluble immune suppressor supernatant (SISS) derived from Con-A-activated mononuclear cell cultures. PHA responses were inhibited by 80 and 95% with SISS and WGA, respectively. The inhibition by both WGA and SISS was totally reversed with addition of GluNAc. Furthermore, WGA and SISS demonstrated competition for the same cell surface receptor site. WGA may therefore be useful as an in vitro model of a saccharide-specific, biologically relevant, soluble mediator for the investigation of mechanisms of immunologic suppression.     

Mutual Exclusivity of Hyaluronan and Hyaluronidase in Invasive Group A Streptococcus

A recent analysis of group A Streptococcus (GAS) invasive infections in Australia has shown a predominance of M4 GAS, a serotype recently reported to lack the antiphagocytic hyaluronic acid (HA) capsule. Here, we use molecular genetics and bioinformatics techniques to characterize 17 clinical M4 isolates associated with invasive disease in children during this recent epidemiology. All M4 isolates lacked HA capsule, and whole genome sequence analysis of two isolates revealed the complete absence of the hasABC capsule biosynthesis operon. Conversely, M4 isolates possess a functional HA-degrading hyaluronate lyase (HylA) enzyme that is rendered nonfunctional in other GAS through a point mutation. Transformation with a plasmid expressing hasABC restored partial encapsulation in wild-type (WT) M4 GAS, and full encapsulation in an isogenic M4 mutant lacking HylA. However, partial encapsulation reduced binding to human complement regulatory protein C4BP, did not enhance survival in whole human blood, and did not increase virulence of WT M4 GAS in a mouse model of systemic infection. Bioinformatics analysis found no hasABC homologs in closely related species, suggesting that this operon was a recent acquisition. These data showcase a mutually exclusive interaction of HA capsule and active HylA among strains of this leading human pathogen.     

Mobility is related to species traits in noctuid moths

1. Mobility is important for the understanding of how species survive in fragmented landscapes and cope with increasing rates of habitat and climate change. However, mobility is a difficult trait to explore and is poorly known in most taxa. Species traits have been studied in relation to range shifts, extinction risks, and responses to habitat area and isolation, and have also been suggested as good estimators of mobility. Here we explore the relation between mobility and species traits in noctuid moths. 2. We sampled noctuid moths by an automatic light‐trap on an island far out in the Baltic Sea. We compared traits of the non‐resident species on the island with traits of a species pool of assumed potential migrants from the Swedish mainland. 3. Mobility was significantly related to adult activity period, length of flight period, and the interaction between host‐plant specificity and distribution area. Widely distributed host‐plant generalists were more mobile than host‐plant specialists with more restricted distribution, and species with an adult activity period in August to September moved to the island to a higher extent than species with an adult activity period in May to July. Our results remained qualitatively robust in additional analyses, after controlling for phylogeny and including all species recorded on the island, except for the trait ‘length of flight period’. 4. Our results highlight the importance of the relation between mobility and species traits. Noctuid moths with certain traits move over longer distances than earlier known. This finding is important to include when predicting range dynamics in fragmented and changing landscapes, and when conservation measures of species are devised.     

榆木蠹蛾生物学特性观察

结合田间观察和室内试验,对榆木蠹蛾HolcocerusvicariusWalker的生物学特性进行了较为系统的研究。结果表明,榆木蠹蛾在宁夏3年发生1代,幼虫主要危害枝干和根颈部,幼虫在蛀道内越冬,5月下旬老熟幼虫在被害树周围5—10cm深的沙土内分散化蛹,蛹期（21±5）d。6月初成虫开始出现,有2个羽化高峰,分别为6月中旬和7月下旬,成虫羽化当晚即可交尾,交尾当天或第2天产卵,每雌蛾产卵最多达720粒,卵期（17±5）d,孵化率为72％～88％。未交尾雌雄成虫寿命为5—6d,交尾后雌雄成虫寿命缩短为3—5d。初孵幼虫于6月中旬始见,10月下旬幼虫开始越冬。幼虫孵化后,先危害韧皮部,常10多条聚集在一起,稍大一点即蛀入木质部。本研究为制定切实可行的榆木蠹蛾有效防治提供理论依据。     

温州市第三代二化螟大发生原因分析

第 3代二化螟大发生是由防治第 2代时失去最佳防治适期 ,而使其绝大部分幼虫存活下来 ,成为第 3代有效虫源所致 ;二是发生时间早 ;三是早稻收割期天气差 ,使 2代向 3代的转化率提高。1998温州市第 3代二化螟大发生正是由于上述 3个原因综合的结果 ,发生面积达 13.5万公顷 ,占当年晚稻面积的 95%以上 ,防治面积达 18万公顷 ,为该市近 10年来所少见。     

Les sépultures italiennes du Paléolithique supérieur. Reconstitutions du régime alimentaire

The identification of dietary habits is increasingly seen as a fundamental aspect for studying the ancient human populations. Accordingly, several projects aiming to identify Paleolithic individuals’ dietary patterns were developed to analyze the organic component of bone tissue and identify isotopic markers to reconstruct the food typology. Bone fragments from six individuals were selected for carbon and nitrogen stable isotopes analysis. The interpretation of human isotopic data was framed through a dataset of twenty-one Italian Paleolithic individuals. The isotopic data generated for the Paleolithic individuals agree with the information already provided by the archaeological record concerning the Italian hunter and gatherer communities. Their subsistence economy was essentially grounded upon the exploitation of high protein foods, either from terrestrial fauna resources or inland lacustrine or riverine species.     

A practical validation approach for virus titer testing of avian infectious bursal disease live vaccine according to current regulatory guidelines

The method for virus titer determination of avian infectious bursal disease (IBD) live vaccine, developed long before regulatory validation guidelines is a cell culture based biological assay intended for use in vaccine release testing.The aim of our study was to perform a validation, based on fit-for-purpose principle, of an old 50% tissue culture infectious dose (TCID₅₀) method according to Guidelines of the International Cooperation on Harmonization of Technical Requirements for Registration of Veterinary Medicinal Products (VICH).This paper addresses challenges and discusses some key aspects that should be considered when validating biological methods. A different statistical approach and non-parametric statistics was introduced in validation protocol in order to derive useful information from experimental data. This approach is applicable for a wide range of methods.In conclusion, the previous virus titration method had showed to be precise, accurate, linear, robust and in accordance with current regulatory standards, which indicates that there is no need for additional re-development or upgrades of the method for its suitability for intended use.     

Ab initio maximum likelihood reconstruction from cryo electron microscopy images of an infectious virion of the tailed bacteriophage P22 and maximum likelihood versions of Fourier Shell Correlation appropriate for measuring resolution of spherical or cylindrical objects

A maximum likelihood reconstruction method for an asymmetric reconstruction of the infectious P22 bacteriophage virion is described and demonstrated on a subset of the images used in [Lander, G.C., Tang, L., Casjens, S.R., Gilcrease, E.B., Prevelige, P., Poliakov, A., Potter, C.S., Carragher, B., Johnson, J.E., 2006. The structure of an infectious P22 virion shows the signal for headful DNA packaging. Science 312(5781), 1791–1795]. The method makes no assumptions at any stage regarding the structure of the phage tail or the relative rotational orientation of the phage tail and capsid but rather the structure and the rotation angle are determined as a part of the analysis. A statistical method for determining resolution consistent with maximum likelihood principles based on ideas for cylinders analogous to the ideas for spheres that are embedded in the Fourier Shell Correlation method is described and demonstrated on the P22 reconstruction. With a correlation threshold of .95, the resolution in the tail measured radially is greater than (33.3 Å) and measured axially is greater than (70.6 Å) both with probability p=0.02.     

望天树学名研讨

经考证,新组合的望天树学名Ｓｈｏｒｅａ ｃｈｉｎｅｎｓｉｓ Ｈ．Ｚｈｕ ｃｏｍｂ．ｎｏｖ．是不合法的晚出同名,它的合法正确学名应为Ｓｈｏｒｈａ ｗａｎｇｔｉａｎｓｈｕｅａ Ｙ．Ｋ．Ｙａｎｇ ｅｔ Ｊ．Ｋ．Ｗｕ ｎｏｍ．ｎｏｖ．。对其进行订正的明确目的是：根据现行的《国际植物命名法规》的原则,规则,辅则和规程来解决分类及鉴定中可能被混淆的问题。