Sample records for albopictus cell line

  1. Analysis of the Aedes albopictus C6/36 genome provides insight into cell line utility for viral propagation.

    PubMed

    Miller, Jason R; Koren, Sergey; Dilley, Kari A; Puri, Vinita; Brown, David M; Harkins, Derek M; Thibaud-Nissen, Françoise; Rosen, Benjamin; Chen, Xiao-Guang; Tu, Zhijian; Sharakhov, Igor V; Sharakhova, Maria V; Sebra, Robert; Stockwell, Timothy B; Bergman, Nicholas H; Sutton, Granger G; Phillippy, Adam M; Piermarini, Peter M; Shabman, Reed S

    2018-03-01

    The 50-year-old Aedes albopictus C6/36 cell line is a resource for the detection, amplification, and analysis of mosquito-borne viruses including Zika, dengue, and chikungunya. The cell line is derived from an unknown number of larvae from an unspecified strain of Aedes albopictus mosquitoes. Toward improved utility of the cell line for research in virus transmission, we present an annotated assembly of the C6/36 genome. The C6/36 genome assembly has the largest contig N50 (3.3 Mbp) of any mosquito assembly, presents the sequences of both haplotypes for most of the diploid genome, reveals independent null mutations in both alleles of the Dicer locus, and indicates a male-specific genome. Gene annotation was computed with publicly available mosquito transcript sequences. Gene expression data from cell line RNA sequence identified enrichment of growth-related pathways and conspicuous deficiency in aquaporins and inward rectifier K+ channels. As a test of utility, RNA sequence data from Zika-infected cells were mapped to the C6/36 genome and transcriptome assemblies. Host subtraction reduced the data set by 89%, enabling faster characterization of nonhost reads. The C6/36 genome sequence and annotation should enable additional uses of the cell line to study arbovirus vector interactions and interventions aimed at restricting the spread of human disease.

  2. Cytoarchitecture of Zika virus infection in human neuroblastoma and Aedes albopictus cell lines.

    PubMed

    Offerdahl, Danielle K; Dorward, David W; Hansen, Bryan T; Bloom, Marshall E

    2017-01-15

    The Zika virus (ZIKV) pandemic is a global concern due to its role in the development of congenital anomalies of the central nervous system. This mosquito-borne flavivirus alternates between mammalian and mosquito hosts, but information about the biogenesis of ZIKV is limited. Using a human neuroblastoma cell line (SK-N-SH) and an Aedes albopictus mosquito cell line (C6/36), we characterized ZIKV infection by immunofluorescence, transmission electron microscopy (TEM), and electron tomography (ET) to better understand infection in these disparate host cells. ZIKV replicated well in both cell lines, but infected SK-N-SH cells suffered a lytic crisis. Flaviviruses scavenge host cell membranes to serve as replication platforms and ZIKV showed the hallmarks of this process. Via TEM, we identified virus particles and 60-100nm spherular vesicles. ET revealed these vesicular replication compartments contain smaller 20-30nm spherular structures. Our studies indicate that SK-N-SH and C6/36 cells are relevant models for viral cytoarchitecture study. Published by Elsevier Inc.

  3. Cytoarchitecture of Zika virus infection in human neuroblastoma and Aedes albopictus cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Offerdahl, Danielle K.

    The Zika virus (ZIKV) pandemic is a global concern due to its role in the development of congenital anomalies of the central nervous system. This mosquito-borne flavivirus alternates between mammalian and mosquito hosts, but information about the biogenesis of ZIKV is limited. Using a human neuroblastoma cell line (SK-N-SH) and an Aedes albopictus mosquito cell line (C6/36), we characterized ZIKV infection by immunofluorescence, transmission electron microscopy (TEM), and electron tomography (ET) to better understand infection in these disparate host cells. ZIKV replicated well in both cell lines, but infected SK-N-SH cells suffered a lytic crisis. Flaviviruses scavenge host cell membranesmore » to serve as replication platforms and ZIKV showed the hallmarks of this process. Via TEM, we identified virus particles and 60–100 nm spherular vesicles. ET revealed these vesicular replication compartments contain smaller 20–30 nm spherular structures. Our studies indicate that SK-N-SH and C6/36 cells are relevant models for viral cytoarchitecture study. - Highlights: •First electron tomography of Zika virus cytoarchitecture. •Comparison of Zika virus infection in human neuroblastoma and mosquito cells. •Ultrastructure of Zika virus infection in human neuroblastoma and mosquito cells.« less

  4. Regulation of asparagine-linked oligosaccharide processing. Oligosaccharide processing in Aedes albopictus mosquito cells.

    PubMed

    Hsieh, P; Robbins, P W

    1984-02-25

    We have examined the synthesis and processing of asparagine-linked oligosaccharides from Aedes albopictus C6/36 mosquito cells. These cells synthesized a glucose-containing lipid-linked oligosaccharide with properties identical to that of Glc3Man9GlcNAc2-PP-dolichol. Results of brief pulse label experiments with [3H]mannose were consistent with the transfer of Glc3Man9GlcNAc2 to protein followed by the rapid removal of glucose residues. Pulse-chase experiments established that further processing of oligosaccharides in C6/36 cells resulted in the removal of up to six alpha-linked mannose residues yielding Man3GlcNAc2 whose structure is identical to that of the trimannosyl "core" of N-linked oligosaccharides of vertebrate cells and yeast. Complex-type oligosaccharides were not observed in C6/36 cells. When Sindbis virus was grown in mosquito cells, Man3GlcNAc2 glycans were preferentially located at the two glycosylation sites which were previously shown to have complex glycans in virus grown in vertebrate cells. These Man3GlcNAc2 structures are the most extensively processed oligosaccharides in A. albopictus, and as such, are analogous to the complex glycans of vertebrate cells. We suggest that determinants of oligosaccharide processing which reside in the polypeptide are universally recognized despite evolutionary divergence of the oligosaccharide-processing pathway between insects and vertebrates.

  5. Dual Insect specific virus infection limits Arbovirus replication in Aedes mosquito cells.

    PubMed

    Schultz, Michaela J; Frydman, Horacio M; Connor, John H

    2018-05-01

    Aedes mosquitoes are vectors for many pathogenic viruses. Cell culture systems facilitate the investigation of virus growth in the mosquito vector. We found Zika virus (ZIKV) growth to be consistent in A. albopictus cells but hypervariable in A. aegypti cell lines. As a potential explanation of this variability, we tested the hypothesis that our cells harbored opportunistic viruses. We screened Aedes cell lines for the presence of insect specific viruses (ISVs), Cell-fusing agent virus (CFAV) and Phasi charoen-like virus (PCLV). PCLV was present in the ZIKV-growth-variable A. aegypti cell lines but absent in A. albopictus lines, suggesting that these ISVs may interfere with ZIKV growth. In support of this hypothesis, PCLV infection of CFAV-positive A. albopictus cells inhibited the growth of ZIKV, dengue virus and La Crosse virus. These data suggest ISV infection of cell lines can impact arbovirus growth leading to significant changes in cell permissivity to arbovirus infection. Copyright © 2018 Elsevier Inc. All rights reserved.

  6. wPip Wolbachia contribution to Aedes albopictus SIT performance: Advantages under intensive rearing.

    PubMed

    Puggioli, Arianna; Calvitti, Maurizio; Moretti, Riccardo; Bellini, Romeo

    2016-12-01

    As a part of a project aiming at the suppression of the mosquito vector Aedes albopictus, a specific Ae. albopictus line producing sterile males, ARwP, was tested for its suitability to intense rearing conditions compatible with mass production and field release. This line was developed by the Italian National Agency for New Technologies, Energy and Sustainable Economic Development thanks to the artificial infection with a heterologous Wolbachia strain, resulting in a bidirectional incompatibility pattern with wild-type Ae. albopictus. ARwP was reared under Standard Operating Procedures at the Centro Agricoltura Ambiente and compared with a wild-type strain in terms of time of pupation onset, production of male pupae in the following 24h and mechanical sexing efficacy. Mating competitiveness of ARwP males was also evaluated in comparison with irradiated wild-type males in large field enclosures. ARwP males demonstrated a significantly shorter time of pupation onset, a higher rate of production of male pupae in the following 24h and a lower percentage of residual contaminant females when applying mechanical sexing procedures. In addition, ARwP males were more efficient than wild-types in competing for wild-type females in large enclosures, thus inducing a level of sterility significantly higher than that expected for an equal mating competitiveness. These results encourage the use of this Ae. albopictus strain as suppression tool against Ae. albopictus based on considerations thoroughly discussed in the manuscript. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. piggybac- and PhiC31-Mediated Genetic Transformation of the Asian Tiger Mosquito, Aedes albopictus (Skuse)

    PubMed Central

    Labbé, Geneviève M. C.; Nimmo, Derric D.; Alphey, Luke

    2010-01-01

    Background The Asian tiger mosquito, Aedes albopictus (Skuse), is a vector of several arboviruses including dengue and chikungunya. This highly invasive species originating from Southeast Asia has travelled the world in the last 30 years and is now established in Europe, North and South America, Africa, the Middle East and the Caribbean. In the absence of vaccine or antiviral drugs, efficient mosquito control strategies are crucial. Conventional control methods have so far failed to control Ae. albopictus adequately. Methodology/Principal Findings Germline transformation of Aedes albopictus was achieved by micro-injection of embryos with a piggyBac-based transgene carrying a 3xP3-ECFP marker and an attP site, combined with piggyBac transposase mRNA and piggyBac helper plasmid. Five independent transgenic lines were established, corresponding to an estimated transformation efficiency of 2–3%. Three lines were re-injected with a second-phase plasmid carrying an attB site and a 3xP3-DsRed2 marker, combined with PhiC31 integrase mRNA. Successful site-specific integration was observed in all three lines with an estimated transformation efficiency of 2–6%. Conclusions/Significance Both piggybac- and site-specific PhiC31-mediated germline transformation of Aedes albopictus were successfully achieved. This is the first report of Ae. albopictus germline transformation and engineering, a key step towards studying and controlling this species using novel molecular techniques and genetic control strategies. PMID:20808959

  8. Functional characterization of three MicroRNAs of the Asian Tiger Mosquito, Aedes albopictus

    PubMed Central

    2013-01-01

    Background Temporal and stage specific expression of microRNAs (miRNAs) in embryos, larvae, pupae and adults of Aedes albopictus showed differential expression levels across the four developmental stages, indicating their potential regulatory roles in mosquito development. The functional characterization of these miRNAs was not known. Accordingly our study evaluated the functional characterization of three miRNAs, which are temporally up-regulated in the various developmental stages of Ae. albopictus mosquitoes. Methods miRNA mimics, inhibitors and negative controls were designed and their knock-in and knock-down efficiency were analyzed by qRT-PCR after transfecting the mosquito cell lines C6/36, and also by injecting in their specific developmental stages. The functional role of each individual miRNA was analyzed with various parameters of development such as, hatching rate and hatching time in embryos, eclosion rate in larvae, longevity and fecundity in the adult mosquitoes. Results The knock-in with the specifically designed miRNA mimics showed increased levels of expression of miRNA compared with their normal controls. We confirmed these findings using qRT-PCR, both by in vitro expression in C6/36 mosquito cell lines after transfection as well as in in vivo expression in developmental stages of mosquitoes by microinjection. The knock-down of expression with the corresponding inhibitors showed a considerable decrease in the expression levels of these miRNAs and obvious functional effects in Ae. albopictus development, detected by a decrease in the hatching rate of embryos and eclosion rate in larvae and a marked reduction in longevity and fecundity in adults. Conclusion This study carried out by knock-in and knock-down of specifically and temporally expressed miRNAs in Ae. albopictus by microinjection is a novel study to delineate the importance of the miRNA expression in regulating mosquito development. The knock-down and loss of function of endogenously

  9. Identification of AaCASPS7, an effector caspase in Aedes albopictus.

    PubMed

    Feng, Lingyan; Liu, Hao; Li, Xiaomei; Qiao, Jialu; Wang, Shengya; Guo, Deyin; Liu, Qingzhen

    2016-11-15

    Aedes albopictus mosquito is a vector of various arboviruses and is becoming a significant threat to public health due to its rapid global expansion. Several reports suggest that apoptosis could be a factor limiting arbovirus infection in mosquitoes. Thus, it is significant to identify apoptosis pathway and study the correlation between apoptosis and virus infection in mosquitoes. Apoptosis is a type of programmed cell death that plays a vital role in immunity, development, and tissue homeostasis. Caspases are a family of conserved proteases playing important roles in apoptosis. In this study, we identified Aedes albopictus AaCASPS7, a caspase shared high identity with dipteran insect drICE orthologs. Phylogenetic analysis showed the closest relative of AaCASPS7 was Aedes aegypti AeCASPS7. AaCASPS7 displayed several features that were typical of an effector caspase and showed significant activity to effector caspase substrates. Aacasps7 transcripts were expressed ubiquitously in developmental and adult stages in Aedes albopictus mosquitoes. Transient expression of AaCASPS7 induced caspase-dependent apoptosis in C6/36 cells. Taken together the above data, this study identified a novel caspase, AaCASPS7, which might function as an apoptotic caspase. Further study the function of AaCASPS7 would facilitate better understanding the apoptotic mechanism in Aedes albopictus mosquito. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Invasion of cemeteries in Florida by Aedes albopictus.

    PubMed

    O'Meara, G F; Gettman, A D; Evans, L F; Scheel, F D

    1992-03-01

    Aedes albopictus has been found in 53 of the 67 Florida counties. The initial discoveries in 11 of these counties were made in cemeteries. At several locations, Ae. albopictus became well-established in cemeteries before appearing in nearby accumulations of waste tires. The recycling of plastic floral baskets may be aiding the spread of Ae. albopictus. Mosquitoes were commonly found in all types of flower-holding containers in cemeteries, except bronze vases. In the laboratory, most Aedes aegypti eggs laid in bronze vases hatched, but larvae subsequently died. The spread of Ae. albopictus in cemeteries seems to occur at the expense of Ae. aegypti populations. At one cemetery immature Ae. albopictus and Ae. aegypti were found in about 70% of the Aedes-positive containers at the start of a monitoring program. In subsequent collections from this site, Ae. albopictus was found in nearly all Aedes-positive containers, whereas there was a progressive decrease in containers with Ae. aegypti. This trend did not appear to be the result of any seasonal pattern because in a nearby cemetery where Ae. albopictus was absent, Ae. aegypti did not show a similar decline. Limiting flower-holding containers to those with drain holes or to bronze vases would greatly limit mosquito production.

  11. The Climate Range Expansion of Aedes albopictus (Diptera: Culicidae) in Asia Inferred From the Distribution of Albopictus Subgroup Species of Aedes (Stegomyia).

    PubMed

    Mogi, M; Armbruster, P A; Tuno, N; Aranda, C; Yong, H S

    2017-11-07

    We compared climatic distribution ranges between Aedes albopictus (Skuse) (Diptera: Culicidae) and the five wild (nondomesticated) species of Albopictus Subgroup of Scutellaris Group of Aedes (Stegomyia) in southern Asia. Distribution sites of the wild species concentrate in seasonal forest and savannah climate zones in India, Indochina, and southern China. The distribution of Ae. albopictus is broader than the wild species under 1) tropical rain-forest climate, 2) steppe and temperate savannah climate, and 3) continental climate with large seasonal temperature variation (hot summer and cold winter) at temperate lowlands (northernmost sites 40°N in Ae. albopictus vs 32°N in the wild species). However, the distribution of Ae. albopictus is more limited at tropical and subtropical highlands where the climate is cool but less continental (small seasonal variation, mild summer, and winter). We discuss a possibility that the broader climate ranges of Ae. albopictus are ecological or eco-evolutionary consequences of adaptation to human habitats. We also propose a general scenario for the origin, dispersal, and adaptation of Ae. albopictus in Asia as a hypothesis for future research. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Comprehensive proteome profiling in Aedes albopictus to decipher Wolbachia-arbovirus interference phenomenon.

    PubMed

    Saucereau, Yoann; Valiente Moro, Claire; Dieryckx, Cindy; Dupuy, Jean-William; Tran, Florence-Hélène; Girard, Vincent; Potier, Patrick; Mavingui, Patrick

    2017-08-18

    Aedes albopictus is a vector of arboviruses that cause severe diseases in humans such as Chikungunya, Dengue and Zika fevers. The vector competence of Ae. albopictus varies depending on the mosquito population involved and the virus transmitted. Wolbachia infection status in believed to be among key elements that determine viral transmission efficiency. Little is known about the cellular functions mobilized in Ae. albopictus during co-infection by Wolbachia and a given arbovirus. To decipher this tripartite interaction at the molecular level, we performed a proteome analysis in Ae. albopictus C6/36 cells mono-infected by Wolbachia wAlbB strain or Chikungunya virus (CHIKV), and bi-infected. We first confirmed significant inhibition of CHIKV by Wolbachia. Using two-dimensional gel electrophoresis followed by nano liquid chromatography coupled with tandem mass spectrometry, we identified 600 unique differentially expressed proteins mostly related to glycolysis, translation and protein metabolism. Wolbachia infection had greater impact on cellular functions than CHIKV infection, inducing either up or down-regulation of proteins associated with metabolic processes such as glycolysis and ATP metabolism, or structural glycoproteins and capsid proteins in the case of bi-infection with CHIKV. CHIKV infection inhibited expression of proteins linked with the processes of transcription, translation, lipid storage and miRNA pathways. The results of our proteome profiling have provided new insights into the molecular pathways involved in tripartite Ae. albopictus-Wolbachia-CHIKV interaction and may help defining targets for the better implementation of Wolbachia-based strategies for disease transmission control.

  13. Temporal Patterns of Abundance of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) and Mitochondrial DNA Analysis of Ae. albopictus in the Central African Republic

    PubMed Central

    Kamgang, Basile; Ngoagouni, Carine; Manirakiza, Alexandre; Nakouné, Emmanuel; Paupy, Christophe; Kazanji, Mirdad

    2013-01-01

    The invasive Asian tiger mosquito Aedes albopictus (Diptera: Culicidae) was first reported in central Africa in 2000, in Cameroon, with the indigenous mosquito species Ae. aegypti (Diptera: Culicidae). Today, this invasive species is present in almost all countries of the region, including the Central African Republic (CAR), where it was first recorded in 2009. As invasive species of mosquitoes can affect the distribution of native species, resulting in new patterns of vectors and concomitant risk for disease, we undertook a comparative study early and late in the wet season in the capital and the main cities of CAR to document infestation and the ecological preferences of the two species. In addition, we determined the probable geographical origin of invasive populations of Ae. albopictus with two mitochondrial DNA genes, COI and ND5. Analysis revealed that Ae. aegypti was more abundant earlier in the wet season and Ae. albopictus in the late wet season. Used tyres were the most heavily colonized productive larval habitats for both species in both seasons. The invasive species Ae. albopictus predominated over the resident species at all sites in which the two species were sympatric. Mitochondrial DNA analysis revealed broad low genetic diversity, confirming recent introduction of Ae. albopictus in CAR. Phylogeographical analysis based on COI polymorphism indicated that the Ae. albopictus haplotype in the CAR population segregated into two lineages, suggesting multiple sources of Ae. albopictus. These data may have important implications for vector control strategies in central Africa. PMID:24349596

  14. Female-specific flightless (fsRIDL) phenotype for control of Aedes albopictus.

    PubMed

    Labbé, Geneviève M C; Scaife, Sarah; Morgan, Siân A; Curtis, Zoë H; Alphey, Luke

    2012-01-01

    Aedes albopictus, the Asian tiger mosquito, is a vector of several arboviruses including dengue and chikungunya, and is also a significant nuisance mosquito. It is one of the most invasive of mosquitoes with a relentlessly increasing geographic distribution. Conventional control methods have so far failed to control Ae. albopictus adequately. Novel genetics-based strategies offer a promising alternative or aid towards efficient control of this mosquito. We describe here the isolation, characterisation and use of the Ae. albopictus Actin-4 gene to drive a dominant lethal gene in the indirect flight muscles of Ae. albopictus, thus inducing a conditional female-specific late-acting flightless phenotype. We also show that in this context, the Actin-4 regulatory regions from both Ae. albopictus and Ae. aegypti can be used to provide conditional female-specific flightlessness in either species. With the disease-transmitting females incapacitated, the female flightless phenotype encompasses a genetic sexing mechanism and would be suitable for controlling Ae. albopictus using a male-only release approach as part of an integrated pest management strategy.

  15. Asymmetrical Competition and Patterns of Abundance of Aedes albopictus and Culex pipiens (Diptera: Culicidae)

    PubMed Central

    COSTANZO, KATIE S.; MORMANN, KIMBERLY; JULIANO, STEVEN A.

    2007-01-01

    We tested for competitive advantage among larvae of Aedes albopictus (Skuse) and Culex pipiens L. in a laboratory experiment and determined the frequency and spatial and temporal patterns of co-occurrence in the field in East St. Louis, IL. In a laboratory competition experiment at multiple combined densities of Ae. albopictus and Cx. pipiens larvae, Ae. albopictus survivorship and developmental times were significantly affected by conspecific densities but not by Cx. pipiens densities. In contrast, Cx. pipiens survivorship and developmental times were significantly affected by both conspecific and Ae. albopictus densities. Per capita rate of increase (r′) for Ae. albopictus cohorts declined significantly due to density of conspecifics, but not density of Cx. pipiens. Interspecific competition between Ae. albopictus and Cx. pipiens under these laboratory conditions was strong and asymmetrical, with the effect of Ae. albopictus on Cx. pipiens much stronger than the reverse. In monthly samples from tire sites in East St. Louis, Ae. albopictus was highly seasonal, occurring in relatively low abundance from early May to July and increasing in abundance in August and September. Co-occurrence corresponded to the seasonality of Ae. albopictus, with Cx. pipiens encountering Ae. albopictus in more tires and at higher numbers within a tire, in August and September. Abundance of both species was high in residential areas and was unrelated to overstory cover, total nitrogen, and total phosphorus. Abundance of Ae. albopictus, but not of Cx. pipiens, was positively associated with conductivity. We expect Cx. pipiens to suffer from the effects of interspecific competition in tires in which it encounters Ae. albopictus. Interspecific competition between these species may be of both ecological and medical importance. PMID:16119544

  16. Spatial and temporal patterns of abundance of Aedes aegypti L. (Stegomyia aegypti) and Aedes albopictus (Skuse) [Stegomyia albopictus (Skuse)] in southern Florida.

    PubMed

    Reiskind, M H; Lounibos, L P

    2013-12-01

    Invasion by mosquito vectors of disease may impact the distribution of resident mosquitoes, resulting in novel patterns of vectors and concomitant risk for disease. One example of such an impact is the invasion by Aedes albopictus (Skuse) [Stegomyia albopictus (Skuse)] (Diptera: Culicidae) of North America and this species' interaction with Aedes aegypti L. (Stegomyia aegypti L). We hypothesized that Ae. aegypti would be found in urban, coastal areas that experience hotter and drier conditions, whereas Ae. albopictus would be more commonly found in suburban and rural areas that are cooler and wetter. In addition, we hypothesized that Ae. aegypti would be more abundant early in the wet season, whereas Ae. albopictus would be more abundant later in the wet season. Urban areas were drier, hotter and contained more Ae. aegypti than suburban or rural areas. Aedes aegypti was relatively more abundant early in the wet season, whereas Ae. albopictus was more abundant in both the late wet season and the dry season. The spatial patterns of inter- and intraspecific encounters between these species were also described. The distribution of these mosquitoes is correlated with abiotic conditions, and with temperature, humidity and the relative availability of rain-filled containers. Understanding the ecological determinants of species distribution can provide insight into the biology of these vectors and important information for their appropriate control. © 2012 The Royal Entomological Society.

  17. Is Switzerland Suitable for the Invasion of Aedes albopictus?

    PubMed Central

    Neteler, Markus; Metz, Markus; Rocchini, Duccio; Rizzoli, Annapaola; Flacio, Eleonora; Engeler, Luca; Guidi, Valeria; Lüthy, Peter; Tonolla, Mauro

    2013-01-01

    Background Over the last 30 years, the Asian tiger mosquito, Aedes albopictus, has rapidly spread around the world. The European distribution comprises the Mediterranean basin with a first appearance in Switzerland in 2003. Early identification of the most suitable areas in Switzerland allowing progressive invasion by this species is considered crucial to suggest adequate surveillance and control plans. Methodology/Principal Findings We identified the most suitable areas for invasion and establishment of Ae. albopictus in Switzerland. The potential distribution areas linked to the current climatic suitability were assessed using remotely sensed land surface temperature data recorded by the MODIS satellite sensors. Suitable areas for adult survival and overwintering of diapausing eggs were also identified for future climatic conditions, considering two different climate change scenarios (A1B, A2) for the periods 2020–2049 and 2045–2074. At present, the areas around Lake Geneva in western Switzerland provide suitable climatic conditions for Ae. albopictus. In northern Switzerland, parts of the Rhine valley, around Lake Constance, as well as the surroundings of Lake Neuchâtel, appear to be suitable for the survival at least of adult Ae. albopictus. However, these areas are characterized by winters currently being too cold for survival and development of diapausing eggs. In southern Switzerland, Ae. albopictus is already well-established, especially in the Canton of Ticino. For the years 2020–2049, the predicted possible spread of the tiger mosquito does not differ significantly from its potential current distribution. However, important expansions are obtained if the period is extended to the years 2045–2074, when Ae. albopictus may invade large new areas. Conclusions/Significance Several parts of Switzerland provide suitable climatic conditions for invasion and establishment of Ae. albopictus. The current distribution and rapid spread in other European

  18. [Aedes albopictus in Italy: an underestimated health problem].

    PubMed

    Romi, R

    2001-01-01

    Since 1990, Ae. albopictus has spread in 9 regions and 30 provinces of the country. This species was introduced in Italy in shipments of scrap tires form the USA. In Italy, Ae. albopictus is the major biting pest throughout much of its range and, although there is no evidence that this mosquito is the vector of human disease in the country, the species might be involved in the transmission of some arboviruses which have been reported in the Mediterranean Basin. Aim of this paper is to provide an update on the distribution of Ae. albopictus in Italy and to renew the interest in a problem frequently underestimated. Public health implications after a ten-year presence of the species are also discussed.

  19. Is Switzerland suitable for the invasion of Aedes albopictus [corrected]?

    PubMed

    Neteler, Markus; Metz, Markus; Rocchini, Duccio; Rizzoli, Annapaola; Flacio, Eleonora; Engeler, Luca; Guidi, Valeria; Lüthy, Peter; Tonolla, Mauro

    2013-01-01

    Over the last 30 years, the Asian tiger mosquito, Aedes albopictus, has rapidly spread around the world. The European distribution comprises the Mediterranean basin with a first appearance in Switzerland in 2003. Early identification of the most suitable areas in Switzerland allowing progressive invasion by this species is considered crucial to suggest adequate surveillance and control plans. We identified the most suitable areas for invasion and establishment of Ae. albopictus in Switzerland. The potential distribution areas linked to the current climatic suitability were assessed using remotely sensed land surface temperature data recorded by the MODIS satellite sensors. Suitable areas for adult survival and overwintering of diapausing eggs were also identified for future climatic conditions, considering two different climate change scenarios (A1B, A2) for the periods 2020-2049 and 2045-2074. At present, the areas around Lake Geneva in western Switzerland provide suitable climatic conditions for Ae. albopictus. In northern Switzerland, parts of the Rhine valley, around Lake Constance, as well as the surroundings of Lake Neuchâtel, appear to be suitable for the survival at least of adult Ae. albopictus. However, these areas are characterized by winters currently being too cold for survival and development of diapausing eggs. In southern Switzerland, Ae. albopictus is already well-established, especially in the Canton of Ticino. For the years 2020-2049, the predicted possible spread of the tiger mosquito does not differ significantly from its potential current distribution. However, important expansions are obtained if the period is extended to the years 2045-2074, when Ae. albopictus may invade large new areas. Several parts of Switzerland provide suitable climatic conditions for invasion and establishment of Ae. albopictus. The current distribution and rapid spread in other European countries suggest that the tiger mosquito will colonize new areas in Switzerland

  20. Fipronil as a larvicide against container-inhabiting mosquito, Aedes albopictus.

    USDA-ARS?s Scientific Manuscript database

    In the laboratory, Fipronil was tested against laboratory-reared and field-collected early 4th instar Aedes albopictus larvae. The insecticide was also bioassayed for activity against natural field populations of Ae. albopictus inhabiting one-liter capacity stone-made containers in a cemetery, St. A...

  1. Aedes albopictus in Allende City, Nuevo Léon, Mexico.

    PubMed

    Pesina, H O; Hernandez, R M; Valdez Rodriguez, M A

    2001-12-01

    Nuevo León was the only 1 of the 3 northeastern states of Mexico where Aedes albopictus previously had not been shown to occur. However, of 195 samples of larval Aedes received during 1997 from the State Health Laboratory for identification from the 7th Sanitary District, Allende City, Nuevo León, 53 (27.2%) were identified as Ae. albopictus.

  2. First record in America of Aedes albopictus naturally infected with dengue virus during the 1995 outbreak at Reynosa, Mexico.

    PubMed

    Ibáñez-Bernal, S; Briseño, B; Mutebi, J P; Argot, E; Rodríguez, G; Martínez-Campos, C; Paz, R; de la Fuente-San Román, P; Tapia-Conyer, R; Flisser, A

    1997-10-01

    Mosquito collections were conducted during a dengue outbreak in Reynosa, Tamaulipas, Mexico, July-December 1995. A total of 6694 adult mosquitoes (four genera and nine species) were captured, of which 2986 (78.3% females and 21.7% males) were Aedes albopictus and 2339 (39.7% females and 60.3% males) were Ae.aegypti. These two species comprised 84.2% of the total collection. Specimens were grouped into pools, nearly 50% of them processed for detection of virus by cythopathic effect in C6-36 and VERO cell cultures and by haemagglutination test. Five pools gave positive haemagglutination reactions and were examined by immunofluorescence using monoclonal antibodies to flavivirus and to dengue virus. One pool of ten Ae.albopictus males was positive for dengue virus: serotypes 2 and 3 were identified by serotype-specific monoclonal antibodies and confirmed by RT-PCR. This is the first report of Ae.albopictus naturally infected with dengue virus in America. Also, it is the very first time Ae.albopictus males have been found infected with dengue virus in the wild.

  3. A MIDGUT DIGESTIVE PHOSPHOLIPASE A2 IN LARVAL MOSQUITOES, AEDES ALBOPICTUS AND CULEX QUINQUEFASCIATUS

    USDA-ARS?s Scientific Manuscript database

    Phospholipase A2 (PLA2) is a secretory digestive enzyme that hydrolyzes ester bond at sn-2 position of dietary phospholipids, creating free fatty acid and lysophopholipid. The free fatty acids (arachidonic acid) are absorbed into midgut cells. Aedes albopictus and Culex quinquefasciatus digestive PL...

  4. The effects of larval habitat quality on Aedes albopictus skip oviposition

    USDA-ARS?s Scientific Manuscript database

    Aedes albopictus, an invasive mosquito species that transmits disease-causing pathogens, oviposits in containers in resource-limited habitats. To mitigate larval competition, Ae. albopictus females may choose to distribute eggs from a single gonotrophic cycle among multiple containers through skip o...

  5. Recent and projected future climatic suitability of North America for the Asian tiger mosquito Aedes albopictus.

    PubMed

    Ogden, Nicholas H; Milka, Radojević; Caminade, Cyril; Gachon, Philippe

    2014-12-02

    Since the 1980s, populations of the Asian tiger mosquito Aedes albopictus have become established in south-eastern, eastern and central United States, extending to approximately 40°N. Ae. albopictus is a vector of a wide range of human pathogens including dengue and chikungunya viruses, which are currently emerging in the Caribbean and Central America and posing a threat to North America. The risk of Ae. albopictus expanding its geographic range in North America under current and future climate was assessed using three climatic indicators of Ae. albopictus survival: overwintering conditions (OW), OW combined with annual air temperature (OWAT), and a linear index of precipitation and air temperature suitability expressed through a sigmoidal function (SIG). The capacity of these indicators to predict Ae. albopictus occurrence was evaluated using surveillance data from the United States. Projected future climatic suitability for Ae. albopictus was obtained using output of nine Regional Climate Model experiments (RCMs). OW and OWAT showed >90% specificity and sensitivity in predicting observed Ae. albopictus occurrence and also predicted moderate to high risk of Ae. albopictus invasion in Pacific coastal areas of the Unites States and Canada under current climate. SIG also well predicted observed Ae. albopictus occurrence (ROC area under the curve was 0.92) but predicted wider current climatic suitability in the north-central and north-eastern United States and south-eastern Canada. RCM output projected modest (circa 500 km) future northward range expansion of Ae. albopictus by the 2050s when using OW and OWAT indicators, but greater (600-1000 km) range expansion, particularly in eastern and central Canada, when using the SIG indicator. Variation in future possible distributions of Ae. albopictus was greater amongst the climatic indicators used than amongst the RCM experiments. Current Ae. albopictus distributions were well predicted by simple climatic indicators

  6. Weak Larval Competition Between Two Invasive Mosquitoes Aedes koreicus and Aedes albopictus (Diptera: Culicidae).

    PubMed

    Baldacchino, Frédéric; Arnoldi, Daniele; Lapère, Charlotte; Rosà, Roberto; Montarsi, Fabrizio; Capelli, Gioia; Rizzoli, Annapaola

    2017-09-01

    Aedes (Hulecoeteomyia) koreicus (Edwards) and Aedes (Stegomyia) albopictus (Skuse) are two invasive mosquito species well established in northeastern Italy, and these two species may co-occur in artificial larval habitats such as tires, buckets, drums, and catch basins. Because Ae. albopictus has been shown experimentally to be a superior competitor to several mosquito species, we investigated larval competition between Ae. koreicus and Ae. albopictus using two diet levels (low level and high level) and 10 Ae. albopictus: Ae. koreicus density combination levels (30:0, 60:0, 15:15, 30:30, 10:20, 20:10, 20:40, 40:20, 0:60, and 0:30). A multivariate analysis (MANOVA) demonstrated a significant effect of the density combination on Ae. koreicus survivorship, female development time, and female wing length considered simultaneously in low-level diet and high-level diet treatments. Pairwise comparisons across low-level diet treatments showed a significant reduction of Ae. koreicus survivorship in 20:10 combination treatments (i.e. 20 Ae. albopictus and 10 Ae. koreicus larvae) compared to 10:20, 20:40, and 30:30 combination treatments, while no difference was detected for Ae. albopictus between density combination treatments. Furthermore, Ae. albopictus developed faster than Ae. koreicus regardless of diet and density combination treatments. Our results show weak larval competition between Ae. koreicus and Ae. albopictus with a slight advantage of the latter species. On the other hand, the presence of Ae. albopictus seems to favor the emergence of larger Ae. koreicus females. We suggest that factors such as habitats preferences or seasonal distributions may be determinant for the invasion success of Ae. koreicus. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  7. Establishment of Aedes albopictus (Diptera: Culicidae) in the Florida Keys, 2001-2017.

    PubMed

    Murray, Heidi L; Pruszynski, Catherine A; Leal, Andrea L; Hribar, Lawrence J

    2018-06-23

    The presence of Aedes albopictus (Skuse) was first reported in the Florida Keys in 1993. Despite extensive surveillance, the Florida Keys Mosquito Control District (FKMCD) only collected specimens occasionally on a few islands, some years finding no evidence of the invasive species. In 2013-2017, FKMCD witnessed a sudden increase in population size and geographic extent of Ae. albopictus. Samples of Ae. albopictus have now been identified on 30 different islands in the Florida Keys. Three islands in particular (Key Largo, Big Pine Key, and Stock Island) have produced multiple positive samples during at least 4 of the last 5 yr, suggesting establishment of the invasive species. FKMCD continues to monitor Ae. albopictus throughout the Keys and make extensive efforts to reduce population abundance and geographic extent of this disease vector.

  8. Combining the Sterile Insect Technique with Wolbachia-Based Approaches: II- A Safer Approach to Aedes albopictus Population Suppression Programmes, Designed to Minimize the Consequences of Inadvertent Female Release

    PubMed Central

    Zhang, Dongjing; Lees, Rosemary Susan; Xi, Zhiyong; Gilles, Jeremie R. L.; Bourtzis, Kostas

    2015-01-01

    Due to the absence of a perfect method for mosquito sex separation, the combination of the sterile insect technique and the incompatible insect technique is now being considered as a potentially effective method to control Aedes albopictus. In this present study first we examine the minimum pupal irradiation dose required to induce complete sterility in Wolbachia triple-infected (HC), double-infected (GUA) and uninfected (GT) female Ae. albopictus. The HC line is a candidate for Ae. albopictus population suppression programmes, but due to the risk of population replacement which characterizes this triple infected line, the individuals to be released need to be additionally irradiated. After determining the minimum irradiation dose required for complete female sterility, we test whether sterilization is sufficient to prevent invasion of the triple infection from the HC females into double-infected (GUA) populations. Our results indicate that irradiated Ae. albopictus HC, GUA and GT strain females have decreased fecundity and egg hatch rate when irradiated, inversely proportional to the dose, and the complete sterilization of females can be acquired by pupal irradiation with doses above 28 Gy. PCR-based analysis of F1 and F2 progeny indicate that the irradiated HC females, cannot spread the new Wolbachia wPip strain into a small cage GUA population, released at a 1:5 ratio. Considering the above results, we conclude that irradiation can be used to reduce the risk of population replacement caused by an unintentional release of Wolbachia triple-infected Ae. albopictus HC strain females during male release for population suppression. PMID:26252474

  9. Stable transformation of a mosquito cell line results in extraordinarily high copy numbers of the plasmid.

    PubMed Central

    Monroe, T J; Muhlmann-Diaz, M C; Kovach, M J; Carlson, J O; Bedford, J S; Beaty, B J

    1992-01-01

    Stable incorporation of high copy numbers (greater than 10,000 per cell) of a plasmid vector containing a gene conferring resistance to the antibiotic hygromycin was achieved in a cell line derived from the Aedes albopictus mosquito. Plasmid sequences were readily observed by ethidium bromide staining of cellular DNA after restriction endonuclease digestion and agarose gel electrophoresis. The plasmid was demonstrated by in situ hybridization to be present in large arrays integrated in metaphase chromosomes and in minute and double-minute replicating elements. In one subclone, approximately 60,000 copies of the plasmid were organized in a large array that resembles a chromosome, morphologically and in the segregation of its chromatids during anaphase. The original as well as modified versions of the plasmid were rescued by transformation of Escherichia coli using total cellular DNA. Southern blot analyses of recovered plasmids indicate the presence of mosquito-derived sequences. Images PMID:1631052

  10. Vector competence of Malaysian Aedes albopictus with and without Wolbachia to four dengue virus serotypes.

    PubMed

    Joanne, Sylvia; Vythilingam, Indra; Teoh, Boon-Teong; Leong, Cherng-Shii; Tan, Kim-Kee; Wong, Meng-Li; Yugavathy, Nava; AbuBakar, Sazaly

    2017-09-01

    To determine the susceptibility status of Aedes albopictus with and without Wolbachia to the four dengue virus serotypes. Two newly colonised colonies of Ae. albopictus from the wild were used for the study. One colony was naturally infected with Wolbachia while in the other Wolbachia was removed by tetracycline treatment. Both colonies were orally infected with dengue virus-infected fresh blood meal. Dengue virus load was measured using quantitative RT-PCR at four-time intervals in the salivary glands, midguts and ovaries. Wolbachia did not significantly affect Malaysian Ae. albopictus dengue infection or the dissemination rate for all four dengue virus serotypes. Malaysian Ae. albopictus had the highest replication kinetics for DENV-1 and the highest salivary gland and midgut infection rate for DENV-4. Wolbachia, which naturally exists in Malaysian Ae. albopictus, does not significantly affect dengue virus replication. Malaysian Ae. albopictus is susceptible to dengue virus infections and capable of transmitting dengue virus, especially DENV-1 and DENV-4. Removal of Wolbachia from Malaysian Ae. albopictus would not reduce their susceptibility status. © 2017 John Wiley & Sons Ltd.

  11. Successful overwintering of Aedes albopictus in Germany.

    PubMed

    Pluskota, Björn; Jöst, Artur; Augsten, Xenia; Stelzner, Lilith; Ferstl, Ina; Becker, Norbert

    2016-08-01

    The Asian tiger mosquito (Aedes albopictus) is of great concern to public health authorities due to its vector competence and rapid spread across the globe. In 2015, two large local breeding populations of Ae. albopictus were discovered in southwest Germany. In spring 2016, we were able to demonstrate the first evidence of a successful overwintering in Germany of this originally tropical mosquito species in different research projects. Particularly noteworthy is the successful hatching of diapause eggs of an Italian strain (Calabria), which overwintered successfully in the field in St. Georgen im Schwarzwald (Baden-Wuerttemberg) at 820 m above sea level. Furthermore, within the scope of a larvae monitoring, the first larvae that hatched in the field were detected on the April 09, 2016 in a rain barrel within the Heidelberg population. Our first results show that self-extinction due to an unsuccessful overwintering cannot be assumed for populations of the Asian tiger mosquito which settled in Germany in previous years. The evidence of a successful overwintering of a large number of diapause eggs and the hatching of the first larvae in field conditions opens the control year against Ae. albopictus in southwest Germany.

  12. Role of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) in local dengue epidemics in Taiwan.

    PubMed

    Tsai, Pui-Jen; Teng, Hwa-Jen

    2016-11-09

    Aedes mosquitoes in Taiwan mainly comprise Aedes albopictus and Ae. aegypti. However, the species contributing to autochthonous dengue spread and the extent at which it occurs remain unclear. Thus, in this study, we spatially analyzed real data to determine spatial features related to local dengue incidence and mosquito density, particularly that of Ae. albopictus and Ae. aegypti. We used bivariate Moran's I statistic and geographically weighted regression (GWR) spatial methods to analyze the globally spatial dependence and locally regressed relationship between (1) imported dengue incidences and Breteau indices (BIs) of Ae. albopictus, (2) imported dengue incidences and BI of Ae. aegypti, (3) autochthonous dengue incidences and BI of Ae. albopictus, (4) autochthonous dengue incidences and BI of Ae. aegypti, (5) all dengue incidences and BI of Ae. albopictus, (6) all dengue incidences and BI of Ae. aegypti, (7) BI of Ae. albopictus and human population density, and (8) BI of Ae. aegypti and human population density in 348 townships in Taiwan. In the GWR models, regression coefficients of spatially regressed relationships between the incidence of autochthonous dengue and vector density of Ae. aegypti were significant and positive in most townships in Taiwan. However, Ae. albopictus had significant but negative regression coefficients in clusters of dengue epidemics. In the global bivariate Moran's index, spatial dependence between the incidence of autochthonous dengue and vector density of Ae. aegypti was significant and exhibited positive correlation in Taiwan (bivariate Moran's index = 0.51). However, Ae. albopictus exhibited positively significant but low correlation (bivariate Moran's index = 0.06). Similar results were observed in the two spatial methods between all dengue incidences and Aedes mosquitoes (Ae. aegypti and Ae. albopictus). The regression coefficients of spatially regressed relationships between imported dengue cases and Aedes mosquitoes

  13. New insights into HCV replication in original cells from Aedes mosquitoes.

    PubMed

    Fallecker, Catherine; Caporossi, Alban; Rechoum, Yassine; Garzoni, Frederic; Larrat, Sylvie; François, Olivier; Fender, Pascal; Morand, Patrice; Berger, Imre; Petit, Marie-Anne; Drouet, Emmanuel

    2017-08-22

    The existing literature about HCV association with, and replication in mosquitoes is extremely poor. To fill this gap, we performed cellular investigations aimed at exploring (i) the capacity of HCV E1E2 glycoproteins to bind on Aedes mosquito cells and (ii) the ability of HCV serum particles (HCVsp) to replicate in these cell lines. First, we used purified E1E2 expressing baculovirus-derived HCV pseudo particles (bacHCVpp) so we could investigate their association with mosquito cell lines from Aedes aegypti (Aag-2) and Aedes albopictus (C6/36). We initiated a series of infections of both mosquito cells (Ae aegypti and Ae albopictus) with the HCVsp (Lat strain - genotype 3) and we observed the evolution dynamics of viral populations within cells over the course of infection via next-generation sequencing (NGS) experiments. Our binding assays revealed bacHCVpp an association with the mosquito cells, at comparable levels obtained with human hepatocytes (HepaRG cells) used as a control. In our infection experiments, the HCV RNA (+) were detectable by RT-PCR in the cells between 21 and 28 days post-infection (p.i.). In human hepatocytes HepaRG and Ae aegypti insect cells, NGS experiments revealed an increase of global viral diversity with a selection for a quasi-species, suggesting a structuration of the population with elimination of deleterious mutations. The evolutionary pattern in Ae albopictus insect cells is different (stability of viral diversity and polymorphism). These results demonstrate for the first time that natural HCV could really replicate within Aedes mosquitoes, a discovery which may have major consequences for public health as well as in vaccine development.

  14. The role of environmental variables on Aedes albopictus biology and chikungunya epidemiology

    PubMed Central

    Waldock, Joanna; Chandra, Nastassya L; Lelieveld, Jos; Proestos, Yiannis; Michael, Edwin; Christophides, George; Parham, Paul E

    2013-01-01

    Aedes albopictus is a vector of dengue and chikungunya viruses in the field, along with around 24 additional arboviruses under laboratory conditions. As an invasive mosquito species, Ae. albopictus has been expanding in geographical range over the past 20 years, although the poleward extent of mosquito populations is limited by winter temperatures. Nonetheless, population densities depend on environmental conditions and since global climate change projections indicate increasing temperatures and altered patterns of rainfall, geographic distributions of previously tropical mosquito species may change. Although mathematical models can provide explanatory insight into observed patterns of disease prevalence in terms of epidemiological and entomological processes, understanding how environmental variables affect transmission is possible only with reliable model parameterisation, which, in turn, is obtained only through a thorough understanding of the relationship between mosquito biology and environmental variables. Thus, in order to assess the impact of climate change on mosquito population distribution and regions threatened by vector-borne disease, a detailed understanding (through a synthesis of current knowledge) of the relationship between climate, mosquito biology, and disease transmission is required, but this process has not yet been undertaken for Ae. albopictus. In this review, the impact of temperature, rainfall, and relative humidity on Ae. albopictus development and survival are considered. Existing Ae. albopictus populations across Europe are mapped with current climatic conditions, considering whether estimates of climatic cutoffs for Ae. albopictus are accurate, and suggesting that environmental thresholds must be calibrated according to the scale and resolution of climate model outputs and mosquito presence data. PMID:23916332

  15. Introduction and potential establishment of Aedes albopictus in California in 2001.

    PubMed

    Linthicum, Kenneth J; Kramer, Vicki L; Madon, Minoo B; Fujioka, Kenn

    2003-12-01

    Aedes albopictus was discovered in Los Angeles, California, in June 2001 in a maritime cargo container from China containing a shipment of a commercial plant product known as "Lucky Bamboo" (Dracaena spp.). To keep the plants alive during the ocean transit, they were shipped in 5-8 cm of water, providing an excellent habitat for Ae. albopictus. Mosquito infestations were subsequently detected at 15 nursery distributors of Dracaena in 2 northern and 4 southern California counties. The distribution of the Ae. albopictus infestations was limited to the vicinity of those nursery distributors with documented infestations. Infestations persisted for more than 5 months near some of the nurseries, and eggs were found in ovitraps until mid-November 2001 up to 1,000 m from the original infestation sites. Overwintering Ae. albopictus populations were discovered in April, July, and August 2002 at original infestation sites in Chino, San Bernardino County, and Monterey Park and Rowland Heights, Los Angeles County, respectively. Specimens were found at some sites of overwintering populations until October 2002.

  16. Holding back the tiger: Successful control program protects Australia from Aedes albopictus expansion

    PubMed Central

    Devine, Gregor; Davis, Joseph; Crunkhorn, Bruce; van den Hurk, Andrew; Whelan, Peter; Russell, Richard; Walker, James; Horne, Peter; Ehlers, Gerhard; Ritchie, Scott

    2017-01-01

    Background The Asian tiger mosquito, Aedes albopictus, is an important vector of dengue, chikungunya and Zika viruses and is a highly invasive and aggressive biter. Established populations of this species were first recognised in Australia in 2005 when they were discovered on islands in the Torres Strait, between mainland Australia and Papua New Guinea. A control program was implemented with the original goal of eliminating Ae. albopictus from the Torres Strait. We describe the evolution of management strategies that provide a template for Ae. albopictus control that can be adopted elsewhere. Methodology / Principal findings The control strategy implemented between 2005 and 2008 targeted larval habitats using source reduction, insect-growth regulator and pyrethroid insecticide to control larvae and adults in the containers. However, the infrequency of insecticide reapplication, the continual accumulation and replacement of containers, and imminent re-introduction of mosquitoes through people’s movement from elsewhere compromised the program. Consequently, in 2009 the objective of the program changed from elimination to quarantine, with the goal of preventing Ae albopictus from infesting Thursday and Horn islands, which are the transport hubs connecting the Torres Strait to mainland Australia. However, larval control strategies did not prevent the species establishing on these islands in 2010. Thereafter, an additional strategy adopted by the quarantine program in early 2011 was harborage spraying, whereby the vegetated, well shaded resting sites of adult Ae. albopictus were treated with a residual pyrethroid insecticide. Inclusion of this additional measure led to a 97% decline in Ae. albopictus numbers within two years. In addition, the frequency of container treatment was increased to five weeks between treatments, compared to an average of 8 weeks that occurred in the earlier iterations of the program. By 2015 and 2016, Ae. albopictus populations on the two

  17. Identification of Aadnr1, a novel gene related to innate immunity and apoptosis in Aedes albopictus.

    PubMed

    Li, Xiaomei; Meng, Kun; Qiao, Jialu; Liu, Hao; Zhong, Chunyan; Liu, Qingzhen

    2016-08-01

    Innate immunity and apoptosis play critical roles in defending pathogens in insects. In Drosophila, Dnr1 was reported as a negative regulator of apoptosis and immune deficiency (Imd) pathway which belongs to innate immunity. Aedes albopictus is an important kind of arbovirus vector and becoming a significant threat to public health due to its rapid global expansion. Here we identified an ortholog of dnr1 from A. albopictus, named as Aadnr1. Aadnr1 encoded a putative protein containing an N-terminal FERM domain and a C-terminal RING domain. AaDnr1 shared high identity with dipteran insects Dnr1 orthologs. Phylogenetic analyses showed that the closest relative of AaDnr1 was Aedes aegypti Dnr1. Real-time PCR proved that Aadnr1 mRNA was expressed ubiquitously during developmental and adult stages. Transcriptional levels of Aadnr1 were decreased drastically in C6/36 cells underwent apoptosis induced by Actinomycin D (Act D) treatment. Partial silence of Aadnr1 enhanced Act D-induced caspase activity. When challenged by heat-inactivated E. coli, transcriptional level of Aadnr1 was also decreased dramatically in C6/36 cells. While when C6/36 cells were infected with Sindbis virus TE/GFP, transcriptional level of Aadnr1 was reduced and recovered repeatedly, with an overall decreasing trend. It was also shown in this study that similar to Drosophila Dnr1, RING domain destabilized AaDnr1 protein. Taken together, the study identified an innate immunity and apoptosis related gene Aadnr1 in A. albopictus. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Dengue serotype-specific immune response in Aedes aegypti and Aedes albopictus

    PubMed Central

    Smartt, Chelsea T; Shin, Dongyoung; Alto, Barry W

    2017-01-01

    BACKGROUND Dengue viruses (DENV) are considered one of the most important emerging pathogens and dengue disease is a global health threat. The geographic expansion of dengue viruses has led to co-circulation of all four dengue serotypes making it imperative that new DENV control strategies be devised. OBJECTIVES Here we characterize dengue serotype-specific innate immune responses in Aedes aegypti and Aedes albopictus using DENV from Puerto Rico (PR). METHODS Ae. aegypti and Ae. albopictus were infected with dengue serotype 1 and 2 isolated from Puerto Rico. DENV infected mosquito samples were collected and temporal change in expression of selected innate immune response pathway genes analyzed by quantitative real time PCR. FINDINGS The Toll pathway is involved in anti-dengue response in Ae. aegypti, and Ae. albopictus. Infections with PR DENV- 1 elicited a stronger response from genes of the Toll immune pathway than PR DENV-2 in Ae. aegypti but in infected Ae. albopictus expression of Toll pathway genes tended to be similar between the serotypes. Two genes (a ribosomal S5 protein gene and a nimrod-like gene) from Ae. albopictus were expressed in response to DENV. MAIN CONCLUSIONS These studies revealed a role for antiviral genes in DENV serotype-specific interactions with DENV vectors, demonstrated that infections with DENV-2 can modulate the Toll immune response pathway in Ae. aegypti and elucidated candidate molecules that might be used to interfere with serotype specific vector-virus interactions. PMID:29211244

  19. Dengue serotype-specific immune response in Aedes aegypti and Aedes albopictus.

    PubMed

    Smartt, Chelsea T; Shin, Dongyoung; Alto, Barry W

    2017-12-01

    Dengue viruses (DENV) are considered one of the most important emerging pathogens and dengue disease is a global health threat. The geographic expansion of dengue viruses has led to co-circulation of all four dengue serotypes making it imperative that new DENV control strategies be devised. Here we characterize dengue serotype-specific innate immune responses in Aedes aegypti and Aedes albopictus using DENV from Puerto Rico (PR). Ae. aegypti and Ae. albopictus were infected with dengue serotype 1 and 2 isolated from Puerto Rico. DENV infected mosquito samples were collected and temporal change in expression of selected innate immune response pathway genes analyzed by quantitative real time PCR. The Toll pathway is involved in anti-dengue response in Ae. aegypti, and Ae. albopictus. Infections with PR DENV- 1 elicited a stronger response from genes of the Toll immune pathway than PR DENV-2 in Ae. aegypti but in infected Ae. albopictus expression of Toll pathway genes tended to be similar between the serotypes. Two genes (a ribosomal S5 protein gene and a nimrod-like gene) from Ae. albopictus were expressed in response to DENV. These studies revealed a role for antiviral genes in DENV serotype-specific interactions with DENV vectors, demonstrated that infections with DENV-2 can modulate the Toll immune response pathway in Ae. aegypti and elucidated candidate molecules that might be used to interfere with serotype specific vector-virus interactions.

  20. The phenetic structure of Aedes albopictus.

    PubMed

    Morales Vargas, Ronald Enrique; Phumala-Morales, Noppawan; Tsunoda, Takashi; Apiwathnasorn, Chamnan; Dujardin, Jean-Pierre

    2013-01-01

    The large and recent migrations of the main vector of the Chikungunya virus, Aedes albopictus, raise questions about the epidemiological impact of possible microevolutionary changes in new territories. Quantitative traits are suitable to detect such changes as induced by environmental adaptations, local competition and founder effects. Using landmark-based geometric morphometrics, we compared the size and shape of 22 populations (1572 females) of Ae. albopictus. The collection sites covered five countries around the world, with special emphasis on Asia, which is assumed to be the origin of the vector. Some collections came from places where an important epidemic outbreak of Chikungunya has recently occurred. Quantitative comparisons were based on 18 anatomical landmarks on the wing. To contrast geographic and possible interspecific shape variation, close species were introduced in the sample, namely five Aedes riversi and nine Ae. flavopictus from Japan. The three species had similar size, but they were clearly separated by shape. Within Ae. albopictus, there was general agreement on size variation with the available climatic data. Shape variation was less related to climatic data than to either geography or a known itinerary of past territorial expansion. Thus, two main clusters were distinguished by wing shape variation: the first one composed of the Southeast Asian sample, the second one grouping Japan, Florida, Hawaii and the Reunion Island samples. The Southeast Asian countries, assumed to be at the origin of the geographic expansion of the mosquito, had similar wings and constituted a distinct group where localities clustered into northern and southern localities. Contrasting with this homogeneous group, very distant localities such as United States (US) and Japan shared a common shape pattern. The US Ae. albopictus samples (Hawaii and Florida) were indeed very similar to the Japanese samples, with Florida behaving exactly like a northern Japanese locality

  1. [Infestation status Aedes albopictus and related mosquito-borne infectious disease risk in central urban area in Shanghai].

    PubMed

    Gao, Q; Xiong, C L; Zhou, Y B; Cao, H; Jiang, Q W

    2016-05-01

    To evaluate Aedes albopictus infestation status in the central urban area of Shanghai, and analyze the related epidemic risk of mosquito-borne infectious disease. Consecutive mosquito surveillance was conducted in the green lands and residential areas in the central urban area of Shanghai during 2012-2014, the Aedes albopictus density and its seasonal fluctuation were observed; the sequence of Aedes albopictus in Shanghai was aligned with that in other epidemic area abroad, and the susceptibility of Aedes albopictus to mosquito-borne virus and endemic risk were analyzed. No Aedes aegypti was found in the central urban area of Shanghai. As predominant species in both the residential area and the green lands, the proportion of Aedes albopictus in the residential area was significantly higher than that in the green lands(78.53% vs. 19.99%, χ(2) =15 525.168, P<0.001), and so was the density(11.91, 42.02 pcs/day · site in the residential area vs. 3.65, 2.18, 2.73 pcs/day · site in the green lands, all P value <0.001). In 2014, the density reached 42.02 pcs/day · site and the proportion reached 94.69% in the residential areas. Phylogenetic tree analysis showed that the genetic distance between Aedes albopictus in Shanghai and Aedes albopictus in Africa was quite far. No Aedes aegypti was found in Shanghai and its surrounding areas, while Aedes albopictus infestation in the central urban area of Shanghai was serious. Strict measures should be taken to reduce the Aedes albopictus density for the effective control Zika virus spread.

  2. Indoor-breeding of Aedes albopictus in northern peninsular Malaysia and its potential epidemiological implications.

    PubMed

    Dieng, Hamady; Saifur, Rahman G M; Hassan, Ahmad Abu; Salmah, M R Che; Boots, Michael; Satho, Tomomitsu; Jaal, Zairi; AbuBakar, Sazaly

    2010-07-27

    The mosquito Ae. albopictus is usually adapted to the peri-domestic environment and typically breeds outdoors. However, we observed its larvae in most containers within homes in northern peninsular Malaysia. To anticipate the epidemiological implications of this indoor-breeding, we assessed some fitness traits affecting vectorial capacity during colonization process. Specifically, we examined whether Ae. albopictus exhibits increased survival, gonotrophic activity and fecundity due to the potential increase in blood feeding opportunities. In a series of experiments involving outdoors and indoors breeding populations, we found that Ae. albopictus lives longer in the indoor environment. We also observed increased nighttime biting activity and lifetime fecundity in indoor/domestic adapted females, although they were similar to recently colonized females in body size. Taken together these data suggest that accommodation of Ae. albopictus to indoor/domestic environment may increase its lifespan, blood feeding success, nuisance and thus vectorial capacity (both in terms of increased vector-host contacts and vector population density). These changes in the breeding behavior of Ae. albopictus, a potential vector of several human pathogens including dengue viruses, require special attention.

  3. Interspecific Larval Competition Between Aedes albopictus and Aedes japonicus (Diptera: Culicidae) in Northern Virginia

    PubMed Central

    Armistead, J. S.; Arias, J. R.; Nishimura, N.; Lounibos, L. P.

    2008-01-01

    Aedes albopictus (Skuse) and Aedes japonicus (Theobald) are two of the most recent and widespread invasive mosquito species to have become established in the United States. The two species co-occur in water-filled artificial containers, where crowding and limiting resources are likely to promote inter- or intraspecific larval competition. The performance of northern Virginia populations of Ae. japonicus and Ae. albopictus competing as larvae under field conditions was evaluated. Per capita rates of population increase for each species were estimated, and the effects of species composition and larval density were determined. In water-containing cups provided with oak leaves, Ae. albopictus larvae exhibited a competitive advantage over Ae. japonicus as a consequence of higher survivorship, shorter developmental time, and a significantly higher estimated population growth rate under conditions of interspecific competition. Intraspecific competition constrained population performance of Ae. albopictus significantly more than competition with Ae. japonicus. In the context of the Lotka-Volterra model of competition, these findings suggest competitive exclusion of Ae. japonicus in those habitats where this species co-occurs with Ae. albopictus. PMID:18714861

  4. Interspecific larval competition between Aedes albopictus and Aedes japonicus (Diptera: Culicidae) in northern Virginia.

    PubMed

    Armistead, J S; Arias, J R; Nishimura, N; Lounibos, L P

    2008-07-01

    Aedes albopictus (Skuse) and Aedes japonicus (Theobald) are two of the most recent and widespread invasive mosquito species to have become established in the United States. The two species co-occur in water-filled artificial containers, where crowding and limiting resources are likely to promote inter- or intraspecific larval competition. The performance of northern Virginia populations of Ae. japonicus and Ae. albopictus competing as larvae under field conditions was evaluated. Per capita rates of population increase for each species were estimated, and the effects of species composition and larval density were determined. In water-containing cups provided with oak leaves, Ae. albopictus larvae exhibited a competitive advantage over Ae. japonicus as a consequence of higher survivorship, shorter developmental time, and a significantly higher estimated population growth rate under conditions of interspecific competition. Intraspecific competition constrained population performance of Ae. albopictus significantly more than competition with Ae. japonicus. In the context of the Lotka-Volterra model of competition, these findings suggest competitive exclusion of Ae. japonicus in those habitats where this species co-occurs with Ae. albopictus.

  5. Insecticide resistance status of United States populations of Aedes albopictus and mechanisms involved

    USDA-ARS?s Scientific Manuscript database

    Aedes albopictus (Skuse) is an invasive mosquito that has become an important vector of chikungunya and dengue viruses. Immature Ae. albopictus thrive in backyard household containers that require treatment with larvicides and when adult populations reach pest levels or disease transmission is ongoi...

  6. The global compendium of Aedes aegypti and Ae. albopictus occurrence

    PubMed Central

    Kraemer, Moritz U. G.; Sinka, Marianne E.; Duda, Kirsten A.; Mylne, Adrian; Shearer, Freya M.; Brady, Oliver J.; Messina, Jane P.; Barker, Christopher M.; Moore, Chester G.; Carvalho, Roberta G.; Coelho, Giovanini E.; Van Bortel, Wim; Hendrickx, Guy; Schaffner, Francis; Wint, G. R. William; Elyazar, Iqbal R. F.; Teng, Hwa-Jen; Hay, Simon I.

    2015-01-01

    Aedes aegypti and Ae. albopictus are the main vectors transmitting dengue and chikungunya viruses. Despite being pathogens of global public health importance, knowledge of their vectors’ global distribution remains patchy and sparse. A global geographic database of known occurrences of Ae. aegypti and Ae. albopictus between 1960 and 2014 was compiled. Herein we present the database, which comprises occurrence data linked to point or polygon locations, derived from peer-reviewed literature and unpublished studies including national entomological surveys and expert networks. We describe all data collection processes, as well as geo-positioning methods, database management and quality-control procedures. This is the first comprehensive global database of Ae. aegypti and Ae. albopictus occurrence, consisting of 19,930 and 22,137 geo-positioned occurrence records respectively. Both datasets can be used for a variety of mapping and spatial analyses of the vectors and, by inference, the diseases they transmit. PMID:26175912

  7. The global compendium of Aedes aegypti and Ae. albopictus occurrence

    NASA Astrophysics Data System (ADS)

    Kraemer, Moritz U. G.; Sinka, Marianne E.; Duda, Kirsten A.; Mylne, Adrian; Shearer, Freya M.; Brady, Oliver J.; Messina, Jane P.; Barker, Christopher M.; Moore, Chester G.; Carvalho, Roberta G.; Coelho, Giovanini E.; van Bortel, Wim; Hendrickx, Guy; Schaffner, Francis; Wint, G. R. William; Elyazar, Iqbal R. F.; Teng, Hwa-Jen; Hay, Simon I.

    2015-07-01

    Aedes aegypti and Ae. albopictus are the main vectors transmitting dengue and chikungunya viruses. Despite being pathogens of global public health importance, knowledge of their vectors’ global distribution remains patchy and sparse. A global geographic database of known occurrences of Ae. aegypti and Ae. albopictus between 1960 and 2014 was compiled. Herein we present the database, which comprises occurrence data linked to point or polygon locations, derived from peer-reviewed literature and unpublished studies including national entomological surveys and expert networks. We describe all data collection processes, as well as geo-positioning methods, database management and quality-control procedures. This is the first comprehensive global database of Ae. aegypti and Ae. albopictus occurrence, consisting of 19,930 and 22,137 geo-positioned occurrence records respectively. Both datasets can be used for a variety of mapping and spatial analyses of the vectors and, by inference, the diseases they transmit.

  8. The association of Aedes aegypti and Ae. albopictus in Allende, Nuevo León, Mexico.

    PubMed

    Mercado-Hernandez, Roberto; Aguilar-Gueta, Juan de Dios; Fernandez-Salas, Ildefoso; Earl, Paul R

    2006-03-01

    The recent appearance of Aedes Stegomyia albopictus (Skuse) in Nuevo León (NL) worries health officials. It is a vector of dengue fever in Asia and is more resistant to lower temperatures than Ae. aegypti. The objective of this study was to learn about some ecological parameters of Ae. albopictus and their association with Ae. aegypti, and other culicids in Allende, NL, Mexico, during 1999. Allende is a small town close to metropolitan Monterrey, which has 4 million inhabitants. The design was random with monthly sampling of 175 ovitraps. Chi-square analyses were performed with data of presence, absence, frequency, and relative abundances. During the study, the species Culex tarsalis (Coquillet), Cx. thriambus (Dyar), Cx. pipiens (Linnaeus), Cx. coronator (Dyar and Knab), Ae. albopictus, Ae. aegypti, Toxorhynchites rutilus (Coquillet), and Ae. triseriatus (Say) were found. April is the month for large numbers of mosquito species. September had the highest populations in positive ovitraps (66.67%), followed by July (63.27% of traps). Aedes aegypti was the most abundant (65.13%), followed by Ae. albopictus (19.71%). Both Ae. albopictus and Ae. aegypti were found from April until December. Aedes aegypti was more abundant than Ae. albopictus, except in August, when they were similar (chi2 = 0.197, P < 0.05). We found significant association between the presence of both species for every study month (chi2 = 9.837, P < 0.05), with a contingency coefficient of 0.247. September and November were the months having the most mosquitoes in this association. Only considering Ae. albopictus, more were found in ovitraps in July (34.6%), followed by September (33.3%). However, its presence was not significant throughout the year. Of 2 zones, in town and at the river, prevalence indicated that Ae. albopictus preferred the river. This mosquito is in its establishment phase in this area and requires further studies.

  9. Introduction, scenarios for establishment and seasonal activity of Aedes albopictus in The Netherlands.

    PubMed

    Takumi, Katsuhisa; Scholte, Ernst-Jan; Braks, Marieta; Reusken, Chantal; Avenell, David; Medlock, Jolyon M

    2009-04-01

    The Asian tiger mosquito Aedes albopictus was detected for the first time in the Netherlands in the summer of 2005. Aedes albopictus is a competent vector of several human viral diseases, and therefore the recent appearance of the vector is a concern to local public health authorities. In 2006 and 2007, the mosquito was found repeatedly and regularly at Lucky bamboo import companies. To assess whether imported Ae. albopictus could establish to produce subsequent generations in the following years or whether the winter conditions in the Netherlands would prove too cold to allow overwintering of diapausing eggs, predictions were made using a Geographic Information Systems (GIS) model based on January average temperature and the annual precipitation recorded in 2006. Seasonal activity of overwintering Ae. albopictus was estimated for temperate strains based on the weekly average temperature and weekly photoperiod using spring egg hatching thresholds of 10.5 degrees C and 11.25 hours, and egg diapause and adult survival thresholds of 9.5 degrees C and 13.5 hours. The analyses indicate that the climate conditions in the Netherlands over the past 10 years were favorable to allow overwintering of diapausing eggs of temperate strains of Ae. albopictus, particularly in the western coastal region. This region was also the area where adult Ae. albopictus were intercepted inside and surrounding plant glasshouses. The estimated number of weeks elapsing between first egg hatching in spring and the production of diapausing eggs in autumn ranged between 17 and 22 weeks in 2006.

  10. Biological characterization of Aedes albopictus (Diptera: Culicidae) in Argentina: implications for arbovirus transmission.

    PubMed

    Chuchuy, Ailen; Rodriguero, Marcela S; Ferrari, Walter; Ciota, Alexander T; Kramer, Laura D; Micieli, María V

    2018-03-22

    Aedes albopictus (Diptera: Culicidae) is an invasive mosquito, native to Asia, that has expanded its range worldwide. It is considered to be a public health threat as it is a competent vector of viruses of medical importance, including dengue, chikungunya, and Zika. Despite its medical importance there is almost no information on biologically important traits of Ae. albopictus in Argentina. We studied life cycle traits, demographic parameters and analyzed the competence of this mosquito as a virus vector. In addition, we determined the prevalence of Wolbachia strains in Ae. albopictus as a first approach to investigate the potential role of this bacteria in modulating vector competence for arboviruses. We observed low hatch rates of eggs, which led to a negative growth rate. We found that Ae. albopictus individuals were infected with Wolbachia in the F1 but while standard superinfection with wAlbA and wAlbB types was found in 66.7% of the females, 16.7% of the females and 62.5% of the males were single-infected with the wAlbB strain. Finally, despite high levels of infection and dissemination, particularly for chikungunya virus, Ae. albopictus from subtropical Argentina were found to be relatively inefficient vectors for transmission of both chikungunya and dengue viruses.

  11. Experimental studies of susceptibility of Italian Aedes albopictus to Zika virus.

    PubMed

    Di Luca, Marco; Severini, Francesco; Toma, Luciano; Boccolini, Daniela; Romi, Roberto; Remoli, Maria Elena; Sabbatucci, Michela; Rizzo, Caterina; Venturi, Giulietta; Rezza, Giovanni; Fortuna, Claudia

    2016-05-05

    We report a study on vector competence of an Italian population of Aedes albopictus for Zika virus (ZIKV). Ae. albopictus was susceptible to ZIKV infection (infection rate: 10%), and the virus could disseminate and was secreted in the mosquito's saliva (dissemination rate: 29%; transmission rate: 29%) after an extrinsic incubation period of 11 days. The observed vector competence was lower than that of an Ae. aegypti colony tested in parallel.

  12. Recovery of Infectious Pariacoto Virus from cDNA Clones and Identification of Susceptible Cell Lines

    PubMed Central

    Johnson, Karyn N.; Ball, L. Andrew

    2001-01-01

    Pariacoto virus (PaV) is a nodavirus that was recently isolated in Peru from the Southern armyworm, Spodoptera eridania. Virus particles are non enveloped and about 30 nm in diameter and have T=3 icosahedral symmetry. The 3.0-Å crystal structure shows that about 35% of the genomic RNA is icosahedrally ordered, with the RNA forming a dodecahedral cage of 25-nucleotide (nt) duplexes that underlie the inner surface of the capsid. The PaV genome comprises two single-stranded, positive-sense RNAs: RNA1 (3,011 nt), which encodes the 108-kDa catalytic subunit of the RNA-dependent RNA polymerase, and RNA2 (1,311 nt), which encodes the 43-kDa capsid protein precursor α. In order to apply molecular genetics to the structure and assembly of PaV, we identified susceptible cell lines and developed a reverse genetic system for this virus. Cell lines that were susceptible to infection by PaV included those from Spodoptera exigua, Helicoverpa zea and Aedes albopictus, whereas cells from Drosophila melanogaster and Spodoptera frugiperda were refractory to infection. To recover virus from molecular clones, full-length cDNAs of PaV RNAs 1 and 2 were cotranscribed by T7 RNA polymerase in baby hamster kidney cells that expressed T7 RNA polymerase. Lysates of these cells were infectious both for cultured cells from Helicoverpa zea (corn earworm) and for larvae of Galleria mellonella (greater wax moth). The combination of infectious cDNA clones, cell culture infectivity, and the ability to produce milligram amounts of virus allows the application of DNA-based genetic methods to the study of PaV structure and assembly. PMID:11711613

  13. Recovery of infectious pariacoto virus from cDNA clones and identification of susceptible cell lines.

    PubMed

    Johnson, K N; Ball, L A

    2001-12-01

    Pariacoto virus (PaV) is a nodavirus that was recently isolated in Peru from the Southern armyworm, Spodoptera eridania. Virus particles are non enveloped and about 30 nm in diameter and have T=3 icosahedral symmetry. The 3.0-A crystal structure shows that about 35% of the genomic RNA is icosahedrally ordered, with the RNA forming a dodecahedral cage of 25-nucleotide (nt) duplexes that underlie the inner surface of the capsid. The PaV genome comprises two single-stranded, positive-sense RNAs: RNA1 (3,011 nt), which encodes the 108-kDa catalytic subunit of the RNA-dependent RNA polymerase, and RNA2 (1,311 nt), which encodes the 43-kDa capsid protein precursor alpha. In order to apply molecular genetics to the structure and assembly of PaV, we identified susceptible cell lines and developed a reverse genetic system for this virus. Cell lines that were susceptible to infection by PaV included those from Spodoptera exigua, Helicoverpa zea and Aedes albopictus, whereas cells from Drosophila melanogaster and Spodoptera frugiperda were refractory to infection. To recover virus from molecular clones, full-length cDNAs of PaV RNAs 1 and 2 were cotranscribed by T7 RNA polymerase in baby hamster kidney cells that expressed T7 RNA polymerase. Lysates of these cells were infectious both for cultured cells from Helicoverpa zea (corn earworm) and for larvae of Galleria mellonella (greater wax moth). The combination of infectious cDNA clones, cell culture infectivity, and the ability to produce milligram amounts of virus allows the application of DNA-based genetic methods to the study of PaV structure and assembly.

  14. Indoor-Breeding of Aedes albopictus in Northern Peninsular Malaysia and Its Potential Epidemiological Implications

    PubMed Central

    Dieng, Hamady; Saifur, Rahman G. M.; Hassan, Ahmad Abu; Salmah, M. R. Che; Boots, Michael; Satho, Tomomitsu; Jaal, Zairi; AbuBakar, Sazaly

    2010-01-01

    Background The mosquito Ae. albopictus is usually adapted to the peri-domestic environment and typically breeds outdoors. However, we observed its larvae in most containers within homes in northern peninsular Malaysia. To anticipate the epidemiological implications of this indoor-breeding, we assessed some fitness traits affecting vectorial capacity during colonization process. Specifically, we examined whether Ae. albopictus exhibits increased survival, gonotrophic activity and fecundity due to the potential increase in blood feeding opportunities. Methodology/Principal Findings In a series of experiments involving outdoors and indoors breeding populations, we found that Ae. albopictus lives longer in the indoor environment. We also observed increased nighttime biting activity and lifetime fecundity in indoor/domestic adapted females, although they were similar to recently colonized females in body size. Conclusion/Significance Taken together these data suggest that accommodation of Ae. albopictus to indoor/domestic environment may increase its lifespan, blood feeding success, nuisance and thus vectorial capacity (both in terms of increased vector-host contacts and vector population density). These changes in the breeding behavior of Ae. albopictus, a potential vector of several human pathogens including dengue viruses, require special attention. PMID:20668543

  15. Competence of Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus Mosquitoes as Zika Virus Vectors, China.

    PubMed

    Liu, Zhuanzhuan; Zhou, Tengfei; Lai, Zetian; Zhang, Zhenhong; Jia, Zhirong; Zhou, Guofa; Williams, Tricia; Xu, Jiabao; Gu, Jinbao; Zhou, Xiaohong; Lin, Lifeng; Yan, Guiyun; Chen, Xiao-Guang

    2017-07-01

    In China, the prevention and control of Zika virus disease has been a public health threat since the first imported case was reported in February 2016. To determine the vector competence of potential vector mosquito species, we experimentally infected Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus mosquitoes and determined infection rates, dissemination rates, and transmission rates. We found the highest vector competence for the imported Zika virus in Ae. aegypti mosquitoes, some susceptibility of Ae. albopictus mosquitoes, but no transmission ability for Cx. quinquefasciatus mosquitoes. Considering that, in China, Ae. albopictus mosquitoes are widely distributed but Ae. aegypti mosquito distribution is limited, Ae. albopictus mosquitoes are a potential primary vector for Zika virus and should be targeted in vector control strategies.

  16. Competence of Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus Mosquitoes as Zika Virus Vectors, China

    PubMed Central

    Liu, Zhuanzhuan; Zhou, Tengfei; Lai, Zetian; Zhang, Zhenhong; Jia, Zhirong; Zhou, Guofa; Williams, Tricia; Xu, Jiabao; Gu, Jinbao; Zhou, Xiaohong; Lin, Lifeng; Yan, Guiyun

    2017-01-01

    In China, the prevention and control of Zika virus disease has been a public health threat since the first imported case was reported in February 2016. To determine the vector competence of potential vector mosquito species, we experimentally infected Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus mosquitoes and determined infection rates, dissemination rates, and transmission rates. We found the highest vector competence for the imported Zika virus in Ae. aegypti mosquitoes, some susceptibility of Ae. albopictus mosquitoes, but no transmission ability for Cx. quinquefasciatus mosquitoes. Considering that, in China, Ae. albopictus mosquitoes are widely distributed but Ae. aegypti mosquito distribution is limited, Ae. albopictus mosquitoes are a potential primary vector for Zika virus and should be targeted in vector control strategies. PMID:28430562

  17. The effects of simulated rainfall on immature population dynamics of Aedes albopictus and female oviposition

    NASA Astrophysics Data System (ADS)

    Dieng, Hamady; Rahman, G. M. Saifur; Abu Hassan, A.; Che Salmah, M. R.; Satho, Tomomitsu; Miake, Fumio; Boots, Michael; Sazaly, Abubakar

    2012-01-01

    Larvae of Aedes albopictus Skuse typically inhabit natural and artificial containers. Since these larval habitats are replenished by rainfall, Ae. albopictus may experience increased loss of immature stages in areas with high levels of rainfall. In this study, we investigated the effects of rainfall and container water level on population density, and oviposition activity of Ae. albopictus. In field and laboratory experiments, we found that rainfall resulted in the flushing of breeding habitats. Excess rain negatively impacted larval and pupal retention, especially in small habitats. When filled with water to overflowing, container habitats were significantly repellent to ovipositing females. Taken together, these data suggest that rainfall triggers population loss of Ae. albopictus and related species through a direct detrimental effect (flushing out) and an indirect effect (ovipositional repellency).

  18. Potential Risk Areas of Aedes albopictus in South-Eastern Iran: A Vector of Dengue Fever, Zika, and Chikungunya

    PubMed Central

    Nejati, Jalil; Bueno-Marí, Rubén; Collantes, Francisco; Hanafi-Bojd, Ahmad A.; Vatandoost, Hassan; Charrahy, Zabihollah; Tabatabaei, Seyed M.; Yaghoobi-Ershadi, Mohammad R.; Hasanzehi, Abdolghafar; Shirzadi, Mohammad R.; Moosa-Kazemi, Seyed H.; Sedaghat, Mohammad M.

    2017-01-01

    The possibility of the rapid and global spread of Zika, chikungunya, yellow fever, and dengue fever by Aedes albopictus is well documented and may be facilitated by changes in climate. To avert and manage health risks, climatic and topographic information can be used to model and forecast which areas may be most prone to the establishment of Ae. albopictus. We aimed to weigh and prioritize the predictive value of various meteorological and climatic variables on distributions of Ae. albopictus in south-eastern Iran using the Analytical Hierarchy Process. Out of eight factors used to predict the presence of Ae. albopictus, the highest weighted were land use, followed by temperature, altitude, and precipitation. The inconsistency of this analysis was 0.03 with no missing judgments. The areas predicted to be most at risk of Ae. albopictus-borne diseases were mapped using Geographic Information Systems and remote sensing data. Five-year (2011–2015) meteorological data was collected from 11 meteorological stations and other data was acquired from Landsat and Terra satellite images. Southernmost regions were at greatest risk of Ae. albopictus colonization as well as more urban sites connected by provincial roads. This is the first study in Iran to determine the regional probability of Ae. albopictus establishment. Monitoring and collection of Ae. albopictus from the environment confirmed our projections, though on-going field work is necessary to track the spread of this vector of life-threatening disease. PMID:28928720

  19. Aedes aegypti and Aedes albopictus Habitat Preferences in South Texas, USA

    PubMed Central

    Champion, Samantha R; Vitek, Christopher J

    2014-01-01

    The South Texas region has a historical record of occasional dengue outbreaks. The recent introduction of chikungunya virus to the Caribbean suggests that this disease may be a concern as well. Six different cities and three field habitat types (residential, tire shops, and cemeteries) were examined for evidence of habitat and longitudinal preference of two vector species, Aedes aegypti and Aedes albopictus. A. aegypti was more prevalent in tire shop sites, while A. albopictus was more prevalent in cemetery sites. In residential sites, the relative abundance of the two species varied with longitude, with A. albopictus being more abundant near the coast, and A. aegypti being more abundant inland. There was also a temporal variation, with A. aegypti declining in frequency over time in residential sites. These results have implications for control strategies and disease risk and suggest a greater need for increased surveillance and research in the region. PMID:25520559

  20. Aedes albopictus in the United States: ten-year presence and public health implications.

    PubMed Central

    Moore, C. G.; Mitchell, C. J.

    1997-01-01

    Since its discovery in Houston, Texas, in 1987, the Asian "tiger mosquito" Aedes albopictus has spread to 678 counties in 25 states. This species, which readily colonizes container habitats in the peridomestic environment, was probably introduced into the continental United States in shipments of scrap tires from northern Asia. The early pattern of dispersal followed the interstate highway system, which suggests further dispersal by human activities. The Public Health Service Act of 1988 requires shipments of used tires from countries with Ae. albopictus to be treated to prevent further importations. Given the extensive spread of the mosquito in the United States, it is questionable whether such a requirement is still justified. Ae. albopictus, a major biting pest throughout much of its range, is a competent laboratory vector of at least 22 arboviruses, including many viruses of public health importance. Cache Valley and eastern equine encephalomyelitis viruses are the only human pathogens isolated from U.S. populations of Ae. albopictus. There is no evidence that this mosquito is the vector of human disease in the United States. PMID:9284377

  1. Population genetics and ecological niche of invasive Aedes albopictus in Mexico.

    PubMed

    Pech-May, Angélica; Moo-Llanes, David A; Puerto-Avila, María Belem; Casas, Mauricio; Danis-Lozano, Rogelio; Ponce, Gustavo; Tun-Ku, Ezequiel; Pinto-Castillo, José Francisco; Villegas, Alejandro; Ibáñez-Piñon, Clemente R; González, Cassandra; Ramsey, Janine M

    2016-05-01

    The Asian tiger mosquito Aedes albopictus (Skuse), is one of the most invasive mosquito species worldwide. In Mexico it is now recorded in 12 states and represents a serious public health problem, given the recent introduction of Chikungunya on the southern border. The aim of this study was to analyze the population genetics of A. albopictus from all major recorded foci, and model its ecological niche. Niche similarity with that from its autochthonous distribution in Asia and other invaded countries were analyzed and its potential future expansion and potential human exposure in climate change scenarios measured. We analyzed 125 sequences of a 317 bp fragment of the cyt b gene from seven A. albopictus populations across Mexico. The samples belong to 25 haplotypes with moderate population structuring (Fst=0.081, p<0.02) and population expansion. The most prevalent haplotype, found in all principal sites, was shared with the USA, Brazil, France, Madagascar, and Reunion Island. The ecological niche model using Mexican occurrence records covers 79.7% of the country, and has an 83% overlap with the Asian niche projected to Mexico. Both Neotropical and Nearctic regions are included in the Mexican niche model. Currently in Mexico, 38.6 million inhabitants are exposed to A. albopictus, which is expected to increase to 45.6 million by 2070. Genetic evidence supports collection information that A. albopictus was introduced to Mexico principally by land from the USA and Central and South America. Prevalent haplotypes from Mexico are shared with most invasive regions across the world, just as there was high niche similarity with both natural and invaded regions. The important overlap with the Asian niche model suggests a high potential for the species to disperse to sylvatic regions in Mexico. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Detritus Type Alters the Outcome of Interspecific Competition Between Aedes aegypti and Aedes albopictus (Diptera: Culicidae)

    PubMed Central

    MURRELL, EBONY G.; JULIANO, STEVEN A.

    2008-01-01

    Many studies of interspecific competition between Aedes albopictus (Skuse) and Aedes aegypti (L.) (Diptera: Culicidae) larvae show that Ae. albopictus are superior resource competitors to Ae. aegypti. Single-species studies indicate that growth and survival of Ae. albopictus and Ae. aegypti larvae are affected by the type of detritus present in containers, which presumably affects the amount and quality of microorganisms that the mosquito larvae consume. We tested whether different detritus types alter the intensity of larval competition by raising 10 different density/species combinations of Ae. albopictus and Ae. aegypti larvae under standard laboratory conditions, with one of four detritus types (oak, pine, grass, or insect) provided as a nutrient base. Intraspecific competitive effects on survival were present with all detritus types. Ae. albopictus survivorship was unaffected by interspecific competition in all treatments. Negative interspecific effects on Ae. aegypti survivorship were present with three of four detritus types, but absent with grass. Estimated finite rate of increase (λ’) was lower with pine detritus than with any other detritus type for both species. Furthermore, Ae. aegypti λ’ was negatively affected by high interspecific density in all detritus types except grass. Thus, our experiment confirms competitive asymmetry in favor of Ae. albopictus with oak, pine, or insect detritus, but also demonstrates that certain detritus types may eliminate interspecific competition among the larvae of these species, which may allow for stable coexistence. Such variation in competitive outcome with detritus type may help to account for observed patterns of coexistence/exclusion of Ae. albopictus and Ae. aegypti in the field. PMID:18533429

  3. Age-Stage, Two-Sex Life Table Characteristics of Aedes albopictus and Aedes Aegypti in Penang Island, Malaysia.

    PubMed

    Maimusa, Hamisu A; Ahmad, Abu Hassan; Kassim, Nur Faeza A; Rahim, Junaid

    2016-03-01

    The life table developmental attributes of laboratory colonies of wild strains of Aedes albopictus and Aedes aegypti were analyzed and compared based on the age-stage, two-sex life table. Findings inclusive in this study are: adult preoviposition periods, total preoviposition period, mean intrinsic rate of increase (r), mean finite rate of increase (λ), net reproductive rates (R0), and mean generation time (T). The total preadult development time was 9.47 days for Ae. albopictus and 8.76 days for Ae. aegypti. The life expectancy was 19.01 days for Ae. albopictus and 19.94 days for Ae. aegypti. Mortality occurred mostly during the adult stage. The mean development time for each stage insignificantly correlated with temperature for Ae. albopictus (r  =  -0.208, P > 0.05) and (r  =  -0.312, P > 0.05) for Ae. aegypti. The population parameters suggest that Ae. albopictus and Ae. aegypti populations are r-strategists characterized by a high r, a large R0, and short T. This present study provides the first report to compare the life parameters of Ae. albopictus and Ae. aegypti strains from Penang island, Malaysia.

  4. Crouching Tiger, Hidden Trouble: Urban Sources of Aedes albopictus (Diptera: Culicidae) Refractory to Source-Reduction

    PubMed Central

    Unlu, Isik; Farajollahi, Ary; Strickman, Daniel; Fonseca, Dina M.

    2013-01-01

    Our ultimate objective is to design cost-effective control strategies for Aedes albopictus, the Asian tiger mosquito, an important urban nuisance and disease vector that expanded worldwide during the last 40 years.  We conducted mosquito larval surveys from May through October 2009 in the City of Trenton, New Jersey, USA, while performing intensive monthly source-reduction campaigns that involved removing, emptying, or treating all accessible containers with larvicides and pupicides. We examined patterns of occurrence of Ae. albopictus and Culex pipiens, another urban mosquito, among different container types by comparing observed and expected number of positive containers of each type. Expected use was based on the relative frequency of each container type in the environment. Aedes albopictus larvae and pupae were found significantly more often than expected in medium volumes of water in buckets and plant saucers but were rarely collected in small volumes of water found in trash items such as discarded cups and cans. They were also absent from large volumes of water such as in abandoned swimming pools and catch basins, although we consistently collected Cx. pipiens from those habitats. The frequency of Ae. albopictus in tires indicated rapid and extensive use of these ubiquitous urban containers. Standard larval-based indices did not correlate with adult catches in BG-Sentinel traps, but when based only on Ae. albopictus key containers (buckets, plant saucers, equipment with pockets of water, and tires) they did. Although we found that only 1.2% of the 20,039 water-holding containers examined contained immature Ae. albopictus (5.3% if only key containers were counted), adult populations were still above nuisance action thresholds six times during the 2009 mosquito season. We conclude that in urban New Jersey, effective source reduction for Ae. albopictus control will require scrupulous and repeated cleaning or treatment of everyday use containers and extensive

  5. [Distribution of Aedes albopictus (Skuse) in Nuevo León, México, 2001-2004].

    PubMed

    Orta-Pesina, Héctor; Mercado-Hernández, Roberto; Elizondo-Leal, José Fernando

    2005-01-01

    To determine the distribution of Aedes albopictus (Skuse) in Nuevo Leon between 2001 and 2004. Standardized methods were used to collect 6371 samples of mosquito's larvae from 167 sites. The proportions of samples positive for Ae. albopictus were obtained by municipality and year, using the software program Excel. There were 1179 samples positive for Ae. albopictus. This species was distributed over 10 municipalities, as follows: Five on the Gulf of Mexico Coastal Plains, which predominantly have a semi-warm, sub-humid climate, and precipitations almost year round (Allende, Montemorelos, Linares, General Teran, and Santiago); two (Sabinas Hidalgo, and Cadereyta) localized on the North American Great Plains, which have a warm, semiarid climate, and scarce precipitations. The final three were Apodaca, San Nicolas, and Escobedo distributed over the metropolitan area of Monterrey City. Up to January 2004, the second most important epidemiological vector in the transmission cycle of Ae. albopictus worldwide was found in ten municipalities of Nuevo Leon State, Mexico.

  6. Established Population of the Invasive Mosquito Species Aedes albopictus in Romania, 2012-14.

    PubMed

    Prioteasa, Liviu F; Dinu, Sorin; Fălcuţă, Elena; Ceianu, Cornelia S

    2015-06-01

    During an entomological investigation carried out in Bucharest and surroundings in fall of 2012, 45 adult mosquitoes (38 females and 7 males) of Aedes albopictus were collected in a neighborhood from the southern area of the city. The morphological identification of the species was further confirmed by sequencing 2 mitochondrial DNA markers: the cytochrome c oxidase subunit I and NADH dehydrogenase subunit 5 genes. Aedes albopictus was collected again in 2013 in the same area from July until October. During late summer the species was found also in another location in the city, downtown Bucharest. Larvae were found in water barrels and other types of household containers, as well as in rain catch basins. In 2014, following a nuisance complaint of a Bucharest inhabitant, the entomological investigation found aggressive Ae. albopictus adults on his property that harbored many mosquito larvae in container-type breeding habitats. These findings are the 1st records of this invasive species and of its breeding population in Romania, and show maintenance of the species over 2 winter seasons. Surveillance of the species outside the area of the capital city was not performed, therefore it is not known whether Ae. albopictus has been introduced in other regions of the country. The presence of Ae. albopictus has been reported every year (2012-14) to competent public health authorities, stressing on the importance of surveillance and of implementation of control measures.

  7. [Detection of Aedes albopictus (Skuse) (Diptera: Culicidae) in the city of Cali, Valle del Cauca, Colombia].

    PubMed

    Cuéllar-Jiménez, María Elena; Velásquez-Escobar, Olga Lucía; González-Obando, Ranulfo; Morales-Reichmann, Carlos Andrés

    2007-06-01

    Aedes albopictus is the second most important dengue virus vector in the Asian southeast after Aedes aegypti. Its entrance into the Americas occurred in 1985, and laboratory studies performed show its potential as a vector in this continent as well. In Colombia, this species has been reported in Leticia (Amazonas) in 1998 and Buenaventura (Valle del Cauca) in 2001. The latest discoveries show that this mosquito continues to advance toward the country's interior. To inform that the presence of A. albopictus is documented in the city of Cali, Valle del Cauca, Colombia. Since 2002, weekly sampling has been performed using larval traps located at seventeen stations. The identification of the A. albopictus species, was carried out in the Unidad de Entomología, Laboratorio de Salud Pública Departamental. These identifications were confirmed in the Entomology Laboratory at Universidad del Valle and the National Institute of Health in Bogotá. From April to June of 2006, larvae of A. albopictus were found in six sampling stations located between northwest and northeast of Cali, one of them in the suburban area of the Yumbo city. The control of A. aegypti and A. albopictus must be integrated into a single program. The surveillance in the cities and nearest departments must be intensified with the objective of limiting the advancement of A. albopictus.

  8. A comparative modeling study on non-climatic and climatic risk assessment on Asian Tiger Mosquito (Aedes albopictus)

    PubMed Central

    Shafapour Tehrany, Mahyat; Solhjouy-fard, Samaneh; Kumar, Lalit

    2018-01-01

    Aedes albopictus, the Asian Tiger Mosquito, vector of Chikungunya, Dengue Fever and Zika viruses, has proven its hardy adaptability in expansion from its natural Asian, forest edge, tree hole habitat on the back of international trade transportation, re-establishing in temperate urban surrounds, in a range of water receptacles and semi-enclosures of organic matter. Conventional aerial spray mosquito vector controls focus on wetland and stagnant water expanses, proven to miss the protected hollows and crevices favoured by Ae. albopictus. New control or eradication strategies are thus essential, particular in light of potential expansions in the southeastern and eastern USA. Successful regional vector control strategies require risk level analysis. Should strategies prioritize regions with non-climatic or climatic suitability parameters for Ae. albopictus? Our study used current Ae. albopictus distribution data to develop two independent models: (i) regions with suitable non-climatic factors, and (ii) regions with suitable climate for Ae. albopictus in southeastern USA. Non-climatic model processing used Evidential Belief Function (EBF), together with six geographical conditioning factors (raster data layers), to establish the probability index. Validation of the analysis results was estimated with area under the curve (AUC) using Ae. albopictus presence data. Climatic modeling was based on two General Circulation Models (GCMs), Miroc3.2 and CSIRO-MK30 running the RCP 8.5 scenario in MaxEnt software. EBF non-climatic model results achieved a 0.70 prediction rate and 0.73 success rate, confirming suitability of the study site regions for Ae. albopictus establishment. The climatic model results showed the best-fit model comprised Coldest Quarter Mean Temp, Precipitation of Wettest Quarter and Driest Quarter Precipitation factors with mean AUC value of 0.86. Both GCMs showed that the whole study site is highly suitable and will remain suitable climatically, according to

  9. A comparative modeling study on non-climatic and climatic risk assessment on Asian Tiger Mosquito (Aedes albopictus).

    PubMed

    Shabani, Farzin; Shafapour Tehrany, Mahyat; Solhjouy-Fard, Samaneh; Kumar, Lalit

    2018-01-01

    Aedes albopictus , the Asian Tiger Mosquito, vector of Chikungunya, Dengue Fever and Zika viruses, has proven its hardy adaptability in expansion from its natural Asian, forest edge, tree hole habitat on the back of international trade transportation, re-establishing in temperate urban surrounds, in a range of water receptacles and semi-enclosures of organic matter. Conventional aerial spray mosquito vector controls focus on wetland and stagnant water expanses, proven to miss the protected hollows and crevices favoured by Ae. albopictus. New control or eradication strategies are thus essential, particular in light of potential expansions in the southeastern and eastern USA. Successful regional vector control strategies require risk level analysis. Should strategies prioritize regions with non-climatic or climatic suitability parameters for Ae. albopictus ? Our study used current Ae. albopictus distribution data to develop two independent models: (i) regions with suitable non-climatic factors, and (ii) regions with suitable climate for Ae. albopictus in southeastern USA. Non-climatic model processing used Evidential Belief Function (EBF), together with six geographical conditioning factors (raster data layers), to establish the probability index. Validation of the analysis results was estimated with area under the curve (AUC) using Ae. albopictus presence data. Climatic modeling was based on two General Circulation Models (GCMs), Miroc3.2 and CSIRO-MK30 running the RCP 8.5 scenario in MaxEnt software. EBF non-climatic model results achieved a 0.70 prediction rate and 0.73 success rate, confirming suitability of the study site regions for Ae. albopictus establishment. The climatic model results showed the best-fit model comprised Coldest Quarter Mean Temp, Precipitation of Wettest Quarter and Driest Quarter Precipitation factors with mean AUC value of 0.86. Both GCMs showed that the whole study site is highly suitable and will remain suitable climatically, according

  10. Aedes albopictus in Rome: results and perspectives after 10 years of monitoring.

    PubMed

    Severini, F; Di Luca, M; Toma, L; Romi, R

    2008-06-01

    In 1997, Aedes albopictus (Skuse 1894) was detected in Rome in two opposite areas of the city. In the following 2 years, the species quickly spread. In 2000, scattered foci of the species were reported in the whole urban area and in the outskirts of the capital city. In Rome, Ae. albopictus seems to have found optimal environmental conditions to proliferate and to overwinter through and without diapausing eggs. In ten years Ae. albopictus has colonized the whole urban area through three phases: first massive spread, following maintenance of infestation, and colonization of alternative winter breeding sites with favorable climatic conditions. Data collected during the 2007 show that rainfall is no longer the most important factor for the development of the species, with respect to the past. In fact Ae. albopictus probably has found new alternative larval breeding sites through the colonization of small water collections refilled periodically by human activities. During 2007-2008 winter season, in order to evaluate the species adaptability, a study of eggs hatching and length of larval cycle at low temperatures, was carried out in laboratory and in simulated field conditions. Data and results are showed and discussed also by the light of existing literature.

  11. Chandipura virus growth kinetics in vertebrate cell lines, insect cell lines & embryonated eggs.

    PubMed

    Jadi, R S; Sudeep, A B; Kumar, Satyendra; Arankalle, V A; Mishra, A C

    2010-08-01

    Since not much information on Chandipura virus is available, an attempt was made to study the growth kinetics of the virus in certain vertebrate, invertebrate cell lines and embryonated chicken eggs. Comparative study of Chandipura virus (CHPV) growth kinetics in three vertebrate cell lines [Vero E6, Rhabdo myosarcoma (RD), Porcine stable kidney (PS) cell lines], two insect cell lines [Aedes aegypti (AA) and Phlebotomus papatasi (PP-9) cell lines] and embryonated pathogen free chicken eggs was conducted, by tissue culture infective dose 50 per cent (TCID(50)) and indirect immunofluorescence assay (IFA). All the cell lines and embryonated egg supported the growth of CHPV and yielded high virus titre. The vertebrate cell lines showed distinct cytopathic effect (CPE) within 4-6 h post infection (PI), while no CPE was observed in insect cell lines. PP-9 cell line was the most sensitive system to CHPV as viral antigen could be detected at 1 h PI by IFA. Our results demonstrated that all the systems were susceptible to CHPV and achieved high yield of virus. However, the PP-9 cell line had an edge over the others due to its high sensitivity to the virus which might be useful for detection and isolation of the virus during epidemics.

  12. Comparative studies on Aedes aegypti and Aedes albopictus adult females trespassing commercial nets.

    PubMed

    Andrade, Carlos F S; Cabrini, Isaías

    2010-03-01

    ABSTRACT. The abilities of Aedes aegypti and Ae. albopictus females in trespassing 5 different commercial nets, 2 nets impregnated with deltamethrin or permethrin and 3 non-impregnated nets (Guarany, Perame, and Ricca), were compared. The evaluating parameters were their percent trespassing success, the median trespassing time (TT50), and the relation between mosquito wing sizes and their trespassing success. Some mosquito behavioral traits were also observed. The trespassing success, directly related to opening areas, was found to be lower with impregnated nets for both species. The differences between the mosquito species were significant among all the nets except Ricca. Aedes albopictus showed a very high success rate in trespassing the non-impregnated Perame net and a lower success rate in trespassing Guarany. Aedes albopictus also showed a very high success rate in trespassing pyrethroid-impregnated nets. The TT50 values for the Ricca and Guarany nets were not significantly different between the species, but a significantly lower TT50 value was found for the net with the largest opening (Ricca). The smaller wingspan of Ae. albopictus seems to explain the higher trespassing success of this species.

  13. Repellency of essential oil of Piper aduncum against Aedes albopictus in the laboratory.

    PubMed

    Misni, Norashiqin; Sulaiman, Sallehudin; Othman, Hidayatulfathi; Omar, Baharudin

    2009-12-01

    The repellent activity of Piper aduncum essential oil against Aedes albopictus was investigated under laboratory conditions with human volunteers. The lowest median effective dose (ED50) value was 1.5 microg/cm2 at 60 sec of exposure when compared to 90 sec (2.1 microg/cm2) and 120 sec (1.8 microg/cm2) of exposure. At 0.4 g, the essential oil gave a high protection (95.2%) against Ae. albopictus bites or landing at 2 h postapplication. The percentage of protection was reduced to 83.3% after 4 h, 64.5% after 6 h, and 51.6% after 8 h postapplication. As a comparison, treatment with 10% deet gave 100% protection against mosquito biting/landing for 4 h postapplication. There was no significant difference in percentage protection reduction between the plant extract and the commercial product deet, respectively (P = 0.739). The essential oil, which was not as good as deet, still gave moderate protection against Ae. albopictus biting even until 4 h postapplication. In conclusion, the P. aduncum essential oil has the potential to be used as a repellent against the dengue/dengue hemorrhagic fever vector, Ae. albopictus.

  14. Experimental Transmission of Eastern Equine Encephalitis Virus by Strains of Aedes albopictus and A. taeniorhynchus (Diptera: Culicidae)

    DTIC Science & Technology

    1994-01-01

    AD-A281 335 0 Experimental Transmission of Eastern Equine Encephaliti Vi 4 by Strains of Aedes albopictus and A. taeniorhynch &1j (Diptera: Culicidae...co m •strains of Aedes albopictus (Skuse) was assessed for eastern equine encephalitis (EEE) virus isolated from Ae. albopictus collected in Polk...County, Florida. Both species became infected with and transmitted EEE virus by bite after feeding on 1-d-old chicks that had _been inoculated with EEE

  15. Occurrence and spread in Italy of Aedes albopictus, with implications for its introduction into other parts of Europe.

    PubMed

    Knudsen, A B; Romi, R; Majori, G

    1996-06-01

    Aedes albopictus is considered second only to Ae. aegypti in its importance to man as a disease vector of dengue and dengue hemorrhagic fever. The first sighting in 1979 of the vector species in Europe came from Albania; however, it was only when Ae. albopictus was introduced into Italy in 1990, through the importation of used tires, followed by its subsequent spread, that the species was considered as a threat to public health. At the close of 1995, Ae. albopictus infestations have been reported from 10 Italian regions and 19 provinces. The risk for greater distribution of Ae. albopictus in Europe can potentially be projected, based on well-established criteria such as: where the winter monthly mean temperature is 0 degree C, where at least 50 cm of mean annual rainfall occurs, and where the mean summer temperature is approximately 20 degrees C. Those countries where climatic conditions meet such criteria and that may be vulnerable to a potential introduction of Ae. albopictus include Spain, Portugal, Greece, Turkey, France, Albania, and the former Republic of Yugoslavia. The Italian plan of action, established for the surveillance and control of Ae. albopictus, is presented in detail.

  16. Evaluation of boric acid sugar baits against Aedes albopictus (Diptera: Culicidae) in tropical environments.

    PubMed

    Naranjo, Diana P; Qualls, Whitney A; Müller, Gunter C; Samson, Dayana M; Roque, Deborah; Alimi, Temitope; Arheart, Kristopher; Beier, John C; Xue, Rui-De

    2013-04-01

    Attractive toxic sugar bait (active ingredient, 1% boric acid) was evaluated against Aedes albopictus Skuse populations in the laboratory, semi-field trials, and field trials in residential communities in St. Augustine, Florida. Laboratory evaluations of boric acid sugar baits applied to the plant Pentas lanceolata (Rubiaceae) demonstrated 100 and 92% mortality of A. albopictus at day 7 and 14, respectively. A semi-field study evaluating the bait application to the upperside or topside of leaves resulted in no significant difference on mortality (P>0.05). Overall combined top and bottom boric acid sugar bait application mortality at day 7 was 95% based on leaf bioassays. Field application of the boric acid sugar baits significantly (P<0.05) decreased adult A. albopictus populations up to day 21 post-treatment compared to the pre-treatment population numbers. A significant reduction in oviposition was demonstrated both at day 7 and 14 post-application (P=0.001) as monitored by ovitraps. Attractive toxic sugar bait application in tropical environments demonstrated efficacy, persistence, and feasibility in controlling A. albopictus populations.

  17. CLO: The cell line ontology

    PubMed Central

    2014-01-01

    Background Cell lines have been widely used in biomedical research. The community-based Cell Line Ontology (CLO) is a member of the OBO Foundry library that covers the domain of cell lines. Since its publication two years ago, significant updates have been made, including new groups joining the CLO consortium, new cell line cells, upper level alignment with the Cell Ontology (CL) and the Ontology for Biomedical Investigation, and logical extensions. Construction and content Collaboration among the CLO, CL, and OBI has established consensus definitions of cell line-specific terms such as ‘cell line’, ‘cell line cell’, ‘cell line culturing’, and ‘mortal’ vs. ‘immortal cell line cell’. A cell line is a genetically stable cultured cell population that contains individual cell line cells. The hierarchical structure of the CLO is built based on the hierarchy of the in vivo cell types defined in CL and tissue types (from which cell line cells are derived) defined in the UBERON cross-species anatomy ontology. The new hierarchical structure makes it easier to browse, query, and perform automated classification. We have recently added classes representing more than 2,000 cell line cells from the RIKEN BRC Cell Bank to CLO. Overall, the CLO now contains ~38,000 classes of specific cell line cells derived from over 200 in vivo cell types from various organisms. Utility and discussion The CLO has been applied to different biomedical research studies. Example case studies include annotation and analysis of EBI ArrayExpress data, bioassays, and host-vaccine/pathogen interaction. CLO’s utility goes beyond a catalogue of cell line types. The alignment of the CLO with related ontologies combined with the use of ontological reasoners will support sophisticated inferencing to advance translational informatics development. PMID:25852852

  18. Aedes albopictus in northeast Mexico: an update on adult distribution and first report of parasitism by Ascogregarina taiwanensis.

    PubMed

    Reyes-Villanueva, Filiberto; Garza-Hernande, Javier A; Garcia-Munguia, Alberto M; Howard, Annabel F V; Ortega-Morales, Aldo I; Adeleke, Monsuru A; Rodriguez-Perez, Mario A

    2013-09-01

    Aedes albopictus has been known as efficient vector of dengue in Asian countries and its wide displacement of Ae. aegypti has been documented in many parts of the world. The present survey was carried out to update the distribution of Ae. albopictus in northeast Mexico and to report the first record of parasitism of mosquitoes by Ascogregarina taiwanensis in Mexico. Human landing collections were conducted in the month of May every year between 2007 and 2009 across the three states, Nuevo Leon (NL), Tamaulipas and Coahuila in northeast Mexico. Six human bait collections were also organized at the cemetery of Gomez Farias (GF), a village in southern Tamaulipas during the rainy and dry seasons in 2010. Aedes albopictus caught in 2010 were dissected for parasitic protozoan gregarines. The results of human landing collections carried out during 2007-10 across the three states of northeast zone of Mexico revealed that Ae. albopictus is invading along the route between Monterrey City in NL and Tampico, Tamaulipas, but not into the arid state of Coahuila. Aedes albopictus was recorded in nine new municipalities in addition to the 15 municipalities reported before 2005. Furthermore, six human-bait collections performed during the dry and rainy seasons in 2010 at the cemetery of GF suggest the exclusion of Ae. aegypti on that site. Dominance was shared by Ae. quadrivittatus, another container-inhabitant but indigenous species, and Ae. albopictus during the dry and rainy seasons, respectively. The results of dissection of the mosquitoes for gregarines revealed the parasitism of Ae. albopictus by A. taiwanensis. The results of this study showed that Ae. albopictus has spread to all the municipalities in the northeastern Mexico except the arid area and reported the first record of parasitic protozoan A. taiwanensis in Mexico. We recommend further studies on larval and adult populations of natural container-occupant mosquitoes in northeastern Mexico in order to have a better

  19. Area-wide management of Aedes albopictus: gauging the efficacy of multiple control measures during 2010.

    USDA-ARS?s Scientific Manuscript database

    Aedes albopictus is a major human biting pest and is considered second only to Aedes aegypti in its importance as a disease vector of dengue. Aedes albopictus is responsible for the majority of service requests from urban and suburban residents in New Jersey during recent years; however, standard a...

  20. Characterizing environmental suitability of Aedes albopictus (Diptera: Culicidae) in Mexico based on regional and global niche models.

    PubMed

    Yañez-Arenas, Carlos; Rioja-Nieto, Rodolfo; Martín, Gerardo A; Dzul-Manzanilla, Felipe; Chiappa-Carrara, Xavier; Buenfil-Ávila, Aura; Manrique-Saide, Pablo; Correa-Morales, Fabián; Díaz-Quiñónez, José Alberto; Pérez-Rentería, Crescencio; Ordoñez-Álvarez, José; Vazquez-Prokopec, Gonzalo; Huerta, Herón

    2018-01-10

    The Asian tiger mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae), is an invasive species and a vector of numerous human pathogens, including chikungunya, dengue, yellow fever, and Zika viruses. This mosquito had been reported from 36 geographic locations in Mexico by 2005, increasing to 101 locations by 2010 and 501 locations (spanning 16 states) by 2016. Here we modeled the occupied niche for Ae. albopictus in Mexico to characterize the environmental conditions related to its presence, and to generate updated environmental suitability maps. The predictors with the greatest contribution to characterizing the occupied niche for Ae. albopictus were NDVI and annual mean temperature. We also estimated the environmental suitability for Ae. albopictus in regions of the country where it has not been documented yet, by means of: 1) transferring its occupied niche model to these regions and 2) modeling its fundamental niche using global data. Our models will help vector control and public health institutions to identify areas where Ae. albopictus has not yet been recorded but where it may be present. We emphasize that most of Mexico has environmental conditions that potentially allow the survival of Ae. albopictus, which underscores the need for systematic mosquito monitoring in all states of the country. © The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  1. Dispersion and oviposition of Aedes albopictus in a Brazilian slum: Initial evidence of Asian tiger mosquito domiciliation in urban environments.

    PubMed

    Ayllón, Tania; Câmara, Daniel Cardoso Portela; Morone, Fernanda Cristina; Gonçalves, Larissa da Silva; Saito Monteiro de Barros, Fábio; Brasil, Patrícia; Carvalho, Marilia Sá; Honório, Nildimar Alves

    2018-01-01

    Aedes albopictus, originally considered as a secondary vector for arbovirus transmission, especially in areas where this species co-exist with Aedes aegypti, has been described in most regions of the world. Dispersion and domiciliation of Ae. albopictus in a complex of densely urbanized slums in Rio de Janeiro, Southeastern Brazil, was evidenced. In this study, we tested the hypotheses that 1) Ae. albopictus distribution in urban slums is negatively related to distance from vegetation, and 2) these vectors have taken on a domestic life style with a portion of the population feeding, ovipositing, and resting indoors. To do this, we developed an integrated surveillance proposal, aiming to detect the presence and abundance of Aedes mosquitoes. The study, based on a febrile syndrome surveillance system in a cohort of infants living in the slum complex, was performed on a weekly basis between February 2014 and April 2017. A total of 8,418 adult mosquitoes (3,052 Ae. aegypti, 44 Ae. albopictus, 16 Ae. scapularis, 4 Ae. fluviatilis and 5,302 Culex quinquefasciatus) were collected by direct aspiration and 46,047 Aedes spp. eggs were collected by oviposition traps. The Asian tiger mosquito, Ae. albopictus, was aspirated in its adult form (n = 44), and immature forms of this species (n = 12) were identified from the eggs collected by the ovitraps. In most collection sites, co-occurrence of Ae. aegypti and Ae. albopictus was observed. Key-sites, such as junkyards, thrift stores, factories, tire repair shops and garages, had the higher abundance of Ae. albopictus, followed by schools and households. We collected Ae. albopictus at up to 400 meters to the nearest vegetation cover. The log transformed (n+1) number of females Ae. albopictus captured at each collection point was inversely related to the distance to the nearest vegetation border. These results show that Ae. albopictus, a competent vector for important arboviruses and more commonly found in areas with higher

  2. Dispersion and oviposition of Aedes albopictus in a Brazilian slum: Initial evidence of Asian tiger mosquito domiciliation in urban environments

    PubMed Central

    Câmara, Daniel Cardoso Portela; Morone, Fernanda Cristina; Gonçalves, Larissa da Silva; Saito Monteiro de Barros, Fábio; Brasil, Patrícia; Carvalho, Marilia Sá; Honório, Nildimar Alves

    2018-01-01

    Aedes albopictus, originally considered as a secondary vector for arbovirus transmission, especially in areas where this species co-exist with Aedes aegypti, has been described in most regions of the world. Dispersion and domiciliation of Ae. albopictus in a complex of densely urbanized slums in Rio de Janeiro, Southeastern Brazil, was evidenced. In this study, we tested the hypotheses that 1) Ae. albopictus distribution in urban slums is negatively related to distance from vegetation, and 2) these vectors have taken on a domestic life style with a portion of the population feeding, ovipositing, and resting indoors. To do this, we developed an integrated surveillance proposal, aiming to detect the presence and abundance of Aedes mosquitoes. The study, based on a febrile syndrome surveillance system in a cohort of infants living in the slum complex, was performed on a weekly basis between February 2014 and April 2017. A total of 8,418 adult mosquitoes (3,052 Ae. aegypti, 44 Ae. albopictus, 16 Ae. scapularis, 4 Ae. fluviatilis and 5,302 Culex quinquefasciatus) were collected by direct aspiration and 46,047 Aedes spp. eggs were collected by oviposition traps. The Asian tiger mosquito, Ae. albopictus, was aspirated in its adult form (n = 44), and immature forms of this species (n = 12) were identified from the eggs collected by the ovitraps. In most collection sites, co-occurrence of Ae. aegypti and Ae. albopictus was observed. Key-sites, such as junkyards, thrift stores, factories, tire repair shops and garages, had the higher abundance of Ae. albopictus, followed by schools and households. We collected Ae. albopictus at up to 400 meters to the nearest vegetation cover. The log transformed (n+1) number of females Ae. albopictus captured at each collection point was inversely related to the distance to the nearest vegetation border. These results show that Ae. albopictus, a competent vector for important arboviruses and more commonly found in areas with higher

  3. Northern range expansion of the Asian tiger mosquito (Aedes albopictus): Analysis of mosquito data from Connecticut, USA.

    PubMed

    Armstrong, Philip M; Andreadis, Theodore G; Shepard, John J; Thomas, Michael C

    2017-05-01

    The Asian tiger mosquito (Aedes albopictus) is an invasive species and important arbovirus vector that was introduced into the U.S. in the 1980's where it continues to expand its range. Winter temperature is an important constraint to its northward expansion, with potential range limits located between the 0° and -5°C mean cold month isotherm. Connecticut is located within this climatic zone and therefore, Ae. albopictus was monitored statewide to assess its northern range expansion and to delineate where populations can stably persist. Ae. albopictus females were monitored at fixed trapping sites throughout Connecticut from June-October over a 20-year period, 1997-2016. In addition, Ae. albopictus larvae and pupae were collected from tire habitats and tires were retrieved from the field in the spring and flooded to evaluate overwintering success of hatching larvae. Ae. albopictus was first detected during statewide surveillance when a single adult female was collected in 2006. This species was not collected again until 2010 and was subsequently detected each successive year with increasing abundance and distribution except following the unusually cold winters of 2014 and 2015. Ae. albopictus mosquitoes were most abundant in urban and suburban locations along the southwestern shoreline of Connecticut; however, single specimens were occasionally detected in central parts of the state. Field-collected females were also screened for arbovirus infection yielding two isolations of Cache Valley virus and one isolation of West Nile virus, highlighting the threat posed by this mosquito. Ae. albopictus overwintered in Connecticut under mild winter conditions as shown by recovery of hatched larvae from field collected tires in spring and by early season detection of larvae and pupae. This study documents the establishment and expansion of Ae. albopictus at the northern boundary of its range in the northeastern U.S. and provides a baseline for monitoring the future spread

  4. Northern range expansion of the Asian tiger mosquito (Aedes albopictus): Analysis of mosquito data from Connecticut, USA

    PubMed Central

    Andreadis, Theodore G.; Shepard, John J.; Thomas, Michael C.

    2017-01-01

    Background The Asian tiger mosquito (Aedes albopictus) is an invasive species and important arbovirus vector that was introduced into the U.S. in the 1980's where it continues to expand its range. Winter temperature is an important constraint to its northward expansion, with potential range limits located between the 0° and -5°C mean cold month isotherm. Connecticut is located within this climatic zone and therefore, Ae. albopictus was monitored statewide to assess its northern range expansion and to delineate where populations can stably persist. Methodology/Principal findings Ae. albopictus females were monitored at fixed trapping sites throughout Connecticut from June-October over a 20-year period, 1997–2016. In addition, Ae. albopictus larvae and pupae were collected from tire habitats and tires were retrieved from the field in the spring and flooded to evaluate overwintering success of hatching larvae. Ae. albopictus was first detected during statewide surveillance when a single adult female was collected in 2006. This species was not collected again until 2010 and was subsequently detected each successive year with increasing abundance and distribution except following the unusually cold winters of 2014 and 2015. Ae. albopictus mosquitoes were most abundant in urban and suburban locations along the southwestern shoreline of Connecticut; however, single specimens were occasionally detected in central parts of the state. Field-collected females were also screened for arbovirus infection yielding two isolations of Cache Valley virus and one isolation of West Nile virus, highlighting the threat posed by this mosquito. Ae. albopictus overwintered in Connecticut under mild winter conditions as shown by recovery of hatched larvae from field collected tires in spring and by early season detection of larvae and pupae. Conclusions/Significance This study documents the establishment and expansion of Ae. albopictus at the northern boundary of its range in the

  5. Molecular analysis and genetic diversity of Aedes albopictus (Diptera, Culicidae) from China.

    PubMed

    Ruiling, Zhang; Peien, Leng; Xuejun, Wang; Zhong, Zhang

    2018-05-01

    Aedes albopictus is one of the most invasive species, which can carry Dengue virus, Yellow fever virus and more than twenty arboviruses. Based on mitochondrial gene cytochrome c oxidase I (COI) and samples collected from 17 populations, we investigated the molecular character and genetic diversity of Ae. albopictus from China. Altogether, 25 haplotypes were detected, including 10 shared haplotypes and 15 private haplotypes. H1 was the dominant haplotype, which is widely distributed in 13 populations. Tajima'D value of most populations was significantly negative, demonstrating that populations experienced rapid range expansion recently. Most haplotypes clustered together both in phylogenetic and median-joining network analysis without clear phylogeographic patterns. However, neutrality tests revealed shallow divergences among Hainan and Guangxi with other populations (0.15599 ≤ F ST ≤ 0.75858), which probably due to interrupted gene flow, caused by geographical isolations. In conclusion, Ae. albopictus populations showed low genetic diversity in China.

  6. Detection of Wolbachia in Aedes albopictus and Their Effects on Chikungunya Virus

    PubMed Central

    Ahmad, Noor Afizah; Vythilingam, Indra; Lim, Yvonne A. L.; Zabari, Nur Zatil Aqmar M.; Lee, Han Lim

    2017-01-01

    Wolbachia-based vector control strategies have been proposed as a means to augment the currently existing measures for controlling dengue and chikungunya vectors. Prior to utilizing Wolbachia as a novel vector control strategy, it is crucial to understand the Wolbachia–mosquito interactions. In this study, field surveys were conducted to screen for the infection status of Wolbachia in field-collected Aedes albopictus. The effects of Wolbachia in its native host toward the replication and dissemination of chikungunya virus (CHIKV) was also studied. The prevalence of Wolbachia-infected field-collected Ae. albopictus was estimated to be 98.6% (N = 142) for females and 95.1% (N = 102) for males in the population studied. The Ae. albopictus were naturally infected with both wAlbA and wAlbB strains. We also found that the native Wolbachia has no impact on CHIKV infection and minimal effect on CHIKV dissemination to secondary organs. PMID:27920393

  7. CellLineNavigator: a workbench for cancer cell line analysis

    PubMed Central

    Krupp, Markus; Itzel, Timo; Maass, Thorsten; Hildebrandt, Andreas; Galle, Peter R.; Teufel, Andreas

    2013-01-01

    The CellLineNavigator database, freely available at http://www.medicalgenomics.org/celllinenavigator, is a web-based workbench for large scale comparisons of a large collection of diverse cell lines. It aims to support experimental design in the fields of genomics, systems biology and translational biomedical research. Currently, this compendium holds genome wide expression profiles of 317 different cancer cell lines, categorized into 57 different pathological states and 28 individual tissues. To enlarge the scope of CellLineNavigator, the database was furthermore closely linked to commonly used bioinformatics databases and knowledge repositories. To ensure easy data access and search ability, a simple data and an intuitive querying interface were implemented. It allows the user to explore and filter gene expression, focusing on pathological or physiological conditions. For a more complex search, the advanced query interface may be used to query for (i) differentially expressed genes; (ii) pathological or physiological conditions; or (iii) gene names or functional attributes, such as Kyoto Encyclopaedia of Genes and Genomes pathway maps. These queries may also be combined. Finally, CellLineNavigator allows additional advanced analysis of differentially regulated genes by a direct link to the Database for Annotation, Visualization and Integrated Discovery (DAVID) Bioinformatics Resources. PMID:23118487

  8. Male accessory gland substances from Aedes albopictus affect the locomotor activity of Aedes aegypti females

    PubMed Central

    Lima-Camara, Tamara Nunes; Codeço, Claudia Torres; Honório, Nildimar Alves; Bruno, Rafaela Vieira; Peixoto, Alexandre Afranio; Lounibos, Leon Philip

    2013-01-01

    Dengue is one of the world’s most important mosquito-borne diseases and is usually transmitted by one of two vector species: Aedes aegypti or Aedes albopictus . These two diurnal mosquitoes are frequently found coexisting in similar habitats, enabling interactions between adults, such as cross-mating. The objective of this study was to assess cross-mating between Ae. aegypti females and Ae. albopictus males under artificial conditions and evaluate the locomotor activity of Ae. aegypti virgin females injected with male accessory gland (MAG) homogenates to infer the physiological and behavioural responses to interspecific mating. After seven days of exposure, 3.3-16% of Ae. aegypti females mated with Ae. albopictus males. Virgin Ae. aegypti females injected with conspecific and heterospecific MAGs showed a general decrease in locomotor activity compared to controls and were refractory to mating with conspecific males. The reduction in diurnal locomotor activity induced by injections of conspecific or heterospecific MAGs is consistent with regulation of female reproductive activities by male substances, which are capable of sterilising female Ae. aegypti through satyrisation by Ae. albopictus . PMID:24473799

  9. Aedes albopictus (Diptera: Culicidae) Oviposition Preference as Influenced by Container Size and Buddleja davidii Plants.

    PubMed

    Davis, Timothy J; Kline, Daniel L; Kaufman, Phillip E

    2016-03-01

    Aedes albopictus (Skuse) is a container-breeding mosquito commonly found in residential areas of its range in the United States. Mosquitoes are known to utilize flowering plants for sugar acquisition. Limited information is known about the influences on oviposition site selection, outside of container size. Residential areas are often landscaped with a variety of flowering plants and are known to provide numerous sizes of potential larval developmental sites for container-breeding mosqutioes. Through screened enclosure and field studies, the oviposition preference of Ae. albopictus for containers of three selected sizes (473, 946 and 1,892 ml) and the influence of flowering butterfly bush (Buddleja davidii Franchett cultivar 'Guinevere') plants were examined. Our results document that significantly more eggs were oviposited in the largest containers. Additionally, significantly more eggs were oviposited in containers adjacent to flowering butterfly bushes than in those without a flowering butterfly bush. Finally, our results document that flowering butterfly bushes exerted greater influence over Ae. albopictus oviposition decisions than did container size. Our findings can be applied to several aspects of Ae. albopictus surveillance and control.

  10. Female Adult Aedes albopictus Suppression by Wolbachia-Infected Male Mosquitoes.

    PubMed

    Mains, James W; Brelsfoard, Corey L; Rose, Robert I; Dobson, Stephen L

    2016-09-23

    Dengue, chikungunya and zika viruses are pathogens with an increasing global impact. In the absence of an approved vaccine or therapy, their management relies on controlling the mosquito vectors. But traditional controls are inadequate, and the range of invasive species such as Aedes albopictus (Asian Tiger Mosquito) is expanding. Genetically modified mosquitoes are being tested, but their use has encountered regulatory barriers and public opposition in some countries. Wolbachia bacteria can cause a form of conditional sterility, which can provide an alternative to genetic modification or irradiation. It is unknown however, whether openly released, artificially infected male Ae. albopictus can competitively mate and sterilize females at a level adequate to suppress a field population. Also, the unintended establishment of Wolbachia at the introduction site could result from horizontal transmission or inadvertent female release. In 2014, an Experimental Use Permit from the United States Environmental Protection Agency approved a pilot field trial in Lexington, Kentucky, USA. Here, we present data showing localized reduction of both egg hatch and adult female numbers. The artificial Wolbachia type was not observed to establish in the field. The results are discussed in relation to the applied use of Wolbachia-infected males as a biopesticide to suppress field populations of Ae. albopictus.

  11. Colonized Aedes albopictus and its sexual performance in the wild: implications for SIT technology and containment.

    PubMed

    Hamady, Dieng; Ruslan, Norrafiza Binti; Ahmad, Abu Hassan; Rawi, Che Salmah Md; Ahmad, Hamdan; Satho, Tomomitsu; Miake, Fumio; Zuharah, Wan Fatma; FuKumitsu, Yuki; Saad, Ahmad Ramli; Rajasaygar, Sudha; Vargas, Ronald Enrique Morales; Majid, Abdul Hafiz Ab; Fadzly, Nik; Ghani, Idris Abd; AbuBakar, Sazaly

    2013-07-15

    Mating is a physiological process of crucial importance underlying the size and maintenance of mosquito populations. In sterile and incompatible insect technologies (SIT and IIT), mating is essential for mass production, persistence, and success of released individuals, and is a central parameter for judging the effectiveness of SIT/IIT programs. Some mosquitoes have an enormous reproductive potential for both themselves and pathogens and mating may contribute to persistence of infection in nature. As Aedes albopictus can transmit flaviviruses both sexually and horizontally, and as infected insects are usually derived from laboratory colonies, we investigated the implications of mating between a long-term laboratory colony of Ae. albopictus and wild populations. Through a series of mating experiments, we examined the reproductive outcomes of sexual cross-affinity between laboratory-raised and wild adults of Ae. albopictus. The results indicated appreciable mating compatibility between laboratory-reared and wild adults, and equivalent levels of egg production among reciprocal crosses. We also observed comparable larval eclosion in lab females mated with wild males, and increased adult longevity in female offspring from wild females|×|laboratory males crosses. Taken together, these data suggest that Ae. albopictus can preserve its reproductive fitness over a long period of time in the laboratory environment and has valuable attributes for SIT application. These observations together with the ability to successfully inseminate heterospecific females indicate the potential of Ae. albopictus to act as an ecological barrier if non-sterilized males are massively released in areas occupied by Aedes aegypti. The observed substantial reproductive fitness combined with the capability to reproduce both, itself and viruses illustrates the potential of Ae. albopictus to pose a serious threat if infected and released accidentally.

  12. The invasive Asian tiger mosquito Aedes albopictus (Diptera: Culicidae) in Germany: Local reproduction and overwintering.

    PubMed

    Walther, D; Scheuch, D E; Kampen, H

    2017-02-01

    Within the framework of a German mosquito monitoring programme, the 'Mueckenatlas' (mosquito atlas) has been established as an instrument of citizen participation in mosquito mapping. In 2015, a strikingly large number of Aedes albopictus, which had not been considered established in Germany, was submitted. Three of six collection sites showed local reproduction, with demonstration of developmental stages over three months at two sites. The third populated site was checked only once in October. Developmental stages of Ae. albopictus were found again at these three sites in spring 2016, including one site in southeastern Germany where reproduction had already been documented in 2014. Although population genetic analyses performed on specimens collected at the latter locality in 2014 and 2015 did not provide proof for hibernation, the finding of developmental stages at this and two other very same sites as in the year before and at very early times in the season strongly suggest accomplished overwintering of Ae. albopictus in Germany. Obviously, the second extremely mild winter in Germany in a row and ongoing adaptation of Ae. albopictus to the temperate European climate allow the species to push northwards from endemic regions in the south. Due to the vector competence of Ae. albopictus for numerous pathogens, including dengue, chikungunya and Zika viruses, action should be taken immediately after the detection of local reproduction to eliminate the populations. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Temperature Characterization of Different Urban Microhabitats of Aedes albopictus (Diptera Culicidae) in Central-Northern Italy.

    PubMed

    Vallorani, Roberto; Angelini, Paola; Bellini, Romeo; Carrieri, Marco; Crisci, Alfonso; Mascali Zeo, Silvia; Messeri, Gianni; Venturelli, Claudio

    2015-08-01

    Aedes albopictus (Skuse) is an invasive mosquito species that has spread to many countries in temperate regions bordering the Mediterranean basin, where it is becoming a major public health concern. A good knowledge of the thermal features of the most productive breeding sites for Ae. albopictus is crucial for a better estimation of the mosquitoes' life cycle and developmental rates. In this article, we address the problem of predicting air temperature in three microhabitats common in urban and suburban areas and the air and water temperature inside an ordinary catch basin, which is considered the most productive breeding site for Ae. albopictus in Italy. Temperature differences were statistically proven between the three microhabitats and between the catch basin external and internal temperature. The impacts on the developmental rates for each life stage of Ae. albopictus were tested through a parametric function of the temperature, and the aquatic stages resulted as being the most affected using the specific temperature inside a typical catch basin instead of a generic air temperature. The impact of snow cover on the catch basin internal temperature, and consequently on the mortality of diapausing eggs, was also evaluated. These data can be useful to improve epidemiological models for a better prediction of Ae. albopictus seasonal and population dynamics in central-northern Italian urban areas. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  14. Bioefficacy of Insect Growth Regulators Against Aedes albopictus (Diptera: Culicidea) From Sarawak, Malaysia: A Statewide Survey.

    PubMed

    Lau, Koon Weng; Chen, Chee Dhang; Lee, Han Lim; Low, Van Lun; Sofian-Azirun, Mohd

    2018-05-28

    The susceptibility status of Aedes albopictus (Skuse; Diptera: Culicidea) larvae collected from 13 districts in Sarawak state, Malaysia was evaluated against five insect growth regulators (IGRs) namely, methoprene, pyriproxyfen, diflubenzuron, cyromazine, and novaluron. Field populations of Ae. albopictus were susceptible to methoprene, pyriproxyfen, cyromazine and novaluron with resistance ratios (RRs) ranging from 0.19-0.38, 0.05-0.14, 0.50-0.95, and 0.75-1.00, respectively. Nevertheless, tolerance towards diflubenzuron (0.33-1.33) was observed in this study. In general, these IGRs exhibited promising results and can be used as alternative control agents against field populations of Ae. albopictus in Sarawak, Malaysia.

  15. Detection of the Invasive Mosquito Species Aedes (Stegomyia) albopictus (Diptera: Culicidae) in Portugal.

    PubMed

    Osório, Hugo Costa; Zé-Zé, Líbia; Neto, Maria; Silva, Sílvia; Marques, Fátima; Silva, Ana Sofia; Alves, Maria João

    2018-04-21

    The Asian tiger mosquito Aedes albopictus is an invasive mosquito originating from the Asia-Pacific region. This species is of major concern to public and veterinary health because of its vector role in the transmission of several pathogens, such as chikungunya, dengue, and Zika viruses. In Portugal, a National Vector Surveillance Network (REde de VIgilância de VEctores—REVIVE) is responsible for the surveillance of autochthonous, but also invasive, mosquito species at points of entry, such as airports, ports, storage areas, and specific border regions with Spain. At these locations, networks of mosquito traps are set and maintained under surveillance throughout the year. In September 2017, Ae. albopictus was detected for the first time in a tyre company located in the North of Portugal. Molecular typing was performed, and a preliminary phylogenetic analysis indicated a high similarity with sequences of Ae. albopictus collected in Europe. A prompt surveillance response was locally implemented to determine its dispersal and abundance, and adult mosquitoes were screened for the presence of arboviral RNA. A total of 103 specimens, 52 immatures and 51 adults, were collected. No pathogenic viruses were detected. Despite the obtained results suggest low abundance of the population locally introduced, the risk of dispersal and potential establishment of Ae. albopictus in Portugal has raised concern for autochthonous mosquito-borne disease outbreaks.

  16. Photoperiodic Diapause and the Establishment of Aedes albopictus (Diptera: Culicidae) in North America

    PubMed Central

    2016-01-01

    The invasion and range expansion of Aedes albopictus (Skuse) in North America represents an outstanding opportunity to study processes of invasion, range expansion, and climatic adaptation. Furthermore, knowledge obtained from such research is relevant to developing novel strategies to control this important vector species. Substantial evidence indicates that the photoperiodic diapause response is an important adaptation to climatic variation across the range of Ae. albopictus in North America. Photoperiodic diapause is a key determinant of abundance in both space and time, and the timing of entry into and exit out of diapause strongly affects seasonal population dynamics and thus the potential for arbovirus transmission. Emerging genomic technologies are making it possible to develop high-resolution, genome-wide genetic markers that can be used for genetic mapping of traits relevant to disease transmission and phylogeographic studies to elucidate invasion history. Recent work using next-generation sequencing technologies (e.g., RNA-seq), combined with physiological experiments, has provided extensive insight into the transcriptional basis of the diapause response in Ae. albopictus. Applying this knowledge to identify novel targets for vector control represents an important future challenge. Finally, recent studies have begun to identify traits other than diapause that are affected by photoperiodism. Extending this work to identify additional traits influenced by photoperiod should produce important insights into the seasonal biology of Ae. albopictus. PMID:27354438

  17. Detection of the Invasive Mosquito Species Aedes (Stegomyia) albopictus (Diptera: Culicidae) in Portugal

    PubMed Central

    Osório, Hugo Costa; Zé-Zé, Líbia; Neto, Maria; Silva, Sílvia; Marques, Fátima; Silva, Ana Sofia; Alves, Maria João

    2018-01-01

    The Asian tiger mosquito Aedes albopictus is an invasive mosquito originating from the Asia-Pacific region. This species is of major concern to public and veterinary health because of its vector role in the transmission of several pathogens, such as chikungunya, dengue, and Zika viruses. In Portugal, a National Vector Surveillance Network (REde de VIgilância de VEctores—REVIVE) is responsible for the surveillance of autochthonous, but also invasive, mosquito species at points of entry, such as airports, ports, storage areas, and specific border regions with Spain. At these locations, networks of mosquito traps are set and maintained under surveillance throughout the year. In September 2017, Ae. albopictus was detected for the first time in a tyre company located in the North of Portugal. Molecular typing was performed, and a preliminary phylogenetic analysis indicated a high similarity with sequences of Ae. albopictus collected in Europe. A prompt surveillance response was locally implemented to determine its dispersal and abundance, and adult mosquitoes were screened for the presence of arboviral RNA. A total of 103 specimens, 52 immatures and 51 adults, were collected. No pathogenic viruses were detected. Despite the obtained results suggest low abundance of the population locally introduced, the risk of dispersal and potential establishment of Ae. albopictus in Portugal has raised concern for autochthonous mosquito-borne disease outbreaks. PMID:29690531

  18. Hatch plasticity in response to varied inundation frequency in Aedes albopictus.

    PubMed

    Vitek, Christopher J; Livdahl, Todd

    2009-07-01

    Eggs of container-breeding mosquitoes are able to withstand drought conditions as an egg and hatch when submerged. Frequent rainfall can be simulated by frequent submersion, and drought conditions can be simulated by infrequent submersion. We examined the hatch response of Aedes albopictus (Skuse) eggs to simulated drought conditions. Ae. albopictus eggs from a strain originating outside Kobe, Japan, were subjected to one of three treatments; high-frequency hatch stimulation consisting of submerging the eggs in a nutrient broth mixture every 3 d, low-frequency hatch stimulation consisting of submerging the eggs every 7 d, and delayed high-frequency hatch stimulation. Eggs that were subjected to lower-frequency stimulation showed a significant decrease in hatch delay, which was the opposite of the predicted response. This decrease in hatch delay may be an example of hatch plasticity in response to drought conditions. This response could not be explained as a result of the difference in the ages of the eggs on any given stimulus. A decreased hatch delay response to potential drought conditions combined with rapid larval development may enable Ae. albopictus, whose eggs are not as desiccation resistant as some other container-breeding mosquitoes, to survive extended drought.

  19. Detection of and monitoring for Aedes albopictus (Diptera: Culicidae) in suburban and sylvatic habitats in north central Florida using four sampling techniques.

    PubMed

    Obenauer, P J; Kaufman, P E; Kline, D L; Allan, S A

    2010-10-01

    A sampling study using a BG-Sentinel trap baited with CO(2), a gravid trap baited with an oak-pine infusion, a human subject, and a vegetative aspirator was conducted to compare their reliability at detecting Aedes albopictus Skuse in suburban and sylvatic habitats. We collected 73,849 mosquitoes, representing 29 species from 11 genera over a 20-wk period. The BG-Sentinel trap accounted for over 85% of all Ae. albopictus captured and was significantly more effective at detecting the presence of Ae. albopictus compared with the other three techniques. Landing counts provided the fewest mosquito species (n = 10), yet provided a quick and effective weekly assessment of the major biting species and were the most effective method for sampling Ae. albopictus within a 10-min period. Fewer Ae. albopictus were sampled from sylvatic habitats compared with suburban ones. Sampling criteria advantageous for surveying Ae. albopictus and other mosquito species are discussed.

  20. Analysis of renal cancer cell lines from two major resources enables genomics-guided cell line selection

    NASA Astrophysics Data System (ADS)

    Sinha, Rileen; Winer, Andrew G.; Chevinsky, Michael; Jakubowski, Christopher; Chen, Ying-Bei; Dong, Yiyu; Tickoo, Satish K.; Reuter, Victor E.; Russo, Paul; Coleman, Jonathan A.; Sander, Chris; Hsieh, James J.; Hakimi, A. Ari

    2017-05-01

    The utility of cancer cell lines is affected by the similarity to endogenous tumour cells. Here we compare genomic data from 65 kidney-derived cell lines from the Cancer Cell Line Encyclopedia and the COSMIC Cell Lines Project to three renal cancer subtypes from The Cancer Genome Atlas: clear cell renal cell carcinoma (ccRCC, also known as kidney renal clear cell carcinoma), papillary (pRCC, also known as kidney papillary) and chromophobe (chRCC, also known as kidney chromophobe) renal cell carcinoma. Clustering copy number alterations shows that most cell lines resemble ccRCC, a few (including some often used as models of ccRCC) resemble pRCC, and none resemble chRCC. Human ccRCC tumours clustering with cell lines display clinical and genomic features of more aggressive disease, suggesting that cell lines best represent aggressive tumours. We stratify mutations and copy number alterations for important kidney cancer genes by the consistency between databases, and classify cell lines into established gene expression-based indolent and aggressive subtypes. Our results could aid investigators in analysing appropriate renal cancer cell lines.

  1. Characterization of Bacterial Communities and Asaia Infection with Field-Collected and Laboratory-Reared Aedes albopictus

    DTIC Science & Technology

    2016-08-18

    Characterization of bacterial communities and Asaia infection within field-collected and 1 laboratory-reared Aedes albopictus 2 3 4 Elizabeth S...Running Head: Bacterial communities within Ae. albopictus 10 11 #Address correspondence to Elizabeth S. Andrews, elizabeth.s.andrews11.ctr@mail.mil 12...189 DISTRIBUTION STATEMENT A: Approved for public release; distribution is unlimited. UNCLASSIFIED Abstract 19 The bacterial communities within

  2. First Record of the Asian Tiger Mosquito Aedes albopictus in Hidalgo State, Mexico.

    PubMed

    Ortega-Morales, Aldo I; Cueto-Medina, Sarai M; Rodríguez, Quetzaly K Siller

    2016-09-01

    The occurrence of the Asian tiger mosquito, Aedes albopictus , has been reported in the Mexican states of Tamaulipas, Nuevo León, Coahuila (northeastern), Veracruz, Chiapas, Quintana Roo (southeastern), Morelos, San Luis Potosí (middle), and Sinaloa (northwestern). In April and September 2012, Ae. albopictus was collected in a variety of habitats and landing/biting on the collecting personnel in 12 counties of Hidalgo state (middle). This is the first record of the occurrence of this species in Hidalgo state.

  3. Invasion of Aedes albopictus (Diptera: Culicidae) into central Africa: what consequences for emerging diseases?

    PubMed

    Ngoagouni, Carine; Kamgang, Basile; Nakouné, Emmanuel; Paupy, Chistophe; Kazanji, Mirdad

    2015-03-31

    Aedes albopictus, a mosquito native to Asia, has invaded all five continents during the past three decades. It was reported in central Africa in the 2000s, first in Cameroon, and, since then, has colonised almost all countries of the region. The species, originally considered a secondary vector of dengue viruses, has been showed to play a major role in transmission of chikungunya virus in numerous countries, including in the central African region. We review the current spread of Ae. albopictus in central Africa, its larval ecology and its impact on indigenous species such as Ae. aegypti. We explore the potential of Ae. albopictus to affect the epidemiology of emerging or re-emerging arboviruses and discuss the conventional means for its control, while emphasizing the importance of data on its susceptibility to insecticides to cope with potential outbreaks.

  4. Area-wide management of Aedes albopictus: lessons learned.

    USDA-ARS?s Scientific Manuscript database

    Aedes albopictus, the Asian tiger mosquito, is the principal vector of chikungunya fever and a critical vector of dengue. This daytime biting pest often causes the majority of service requests from urban and suburban residents in New Jersey and many other states and nations where it has spread. Ou...

  5. Increased risk for autochthonous vector-borne infections transmitted by Aedes albopictus in continental Europe.

    PubMed

    Gossner, Céline M; Ducheyne, Els; Schaffner, Francis

    2018-06-01

    Autochthonous outbreaks of chikungunya and dengue during the past decade showed that continental Europe is vulnerable to Aedes albopictus -borne infections. Ae. albopictus has spread geographically, resulting in more people exposed to risk. Timely application of adequate mosquito suppression measures may delay, or even prevent, the vector population from crossing the potential epidemic abundance threshold should a pathogen be introduced. Health authorities should be on alert to detect early cases to prevent autochthonous outbreaks.

  6. Updating the geographical distribution and frequency of Aedes albopictus in Brazil with remarks regarding its range in the Americas

    PubMed Central

    Carvalho, Roberta Gomes; Lourenço-de-Oliveira, Ricardo; Braga, Ima Aparecida

    2014-01-01

    The geographical distribution of Aedes albopictus in Brazil was updated according to the data recorded across the country over the last eight years. Countrywide house indexes (HI) for Ae. albopictus in urban and suburban areas were described for the first time using a sample of Brazilian municipalities. This mosquito is currently present in at least 59% of the Brazilian municipalities and in 24 of the 27 federal units (i.e., 26 states and the Federal District). In 34 Brazilian municipalities, the HI values for Ae. albopictus were higher than those recorded for Ae. aegypti, reaching figures as high as HI = 7.72 in the Southeast Region. Remarks regarding the current range of this mosquito species in the Americas are also presented. Nineteen American countries are currently infested and few mainland American countries have not confirmed the occurrence of Ae. albopictus. The large distribution and high frequency of Ae. albopictus in the Americas may become a critical factor in the spread of arboviruses like chikungunya in the new world. PMID:25317707

  7. Updating the geographical distribution and frequency of Aedes albopictus in Brazil with remarks regarding its range in the Americas.

    PubMed

    Carvalho, Roberta Gomes; Lourenço-de-Oliveira, Ricardo; Braga, Ima Aparecida

    2014-09-01

    The geographical distribution of Aedes albopictus in Brazil was updated according to the data recorded across the country over the last eight years. Countrywide house indexes (HI) for Ae. albopictus in urban and suburban areas were described for the first time using a sample of Brazilian municipalities. This mosquito is currently present in at least 59% of the Brazilian municipalities and in 24 of the 27 federal units (i.e., 26 states and the Federal District). In 34 Brazilian municipalities, the HI values for Ae. albopictus were higher than those recorded for Ae. aegypti, reaching figures as high as HI = 7.72 in the Southeast Region. Remarks regarding the current range of this mosquito species in the Americas are also presented. Nineteen American countries are currently infested and few mainland American countries have not confirmed the occurrence of Ae. albopictus. The large distribution and high frequency of Ae. albopictus in the Americas may become a critical factor in the spread of arboviruses like chikungunya in the new world.

  8. Global temperature constraints on Aedes aegypti and Ae. albopictus persistence and competence for dengue virus transmission

    PubMed Central

    2014-01-01

    Background Dengue is a disease that has undergone significant expansion over the past hundred years. Understanding what factors limit the distribution of transmission can be used to predict current and future limits to further dengue expansion. While not the only factor, temperature plays an important role in defining these limits. Previous attempts to analyse the effect of temperature on the geographic distribution of dengue have not considered its dynamic intra-annual and diurnal change and its cumulative effects on mosquito and virus populations. Methods Here we expand an existing modelling framework with new temperature-based relationships to model an index proportional to the basic reproductive number of the dengue virus. This model framework is combined with high spatial and temporal resolution global temperature data to model the effects of temperature on Aedes aegypti and Ae. albopictus persistence and competence for dengue virus transmission. Results Our model predicted areas where temperature is not expected to permit transmission and/or Aedes persistence throughout the year. By reanalysing existing experimental data our analysis indicates that Ae. albopictus, often considered a minor vector of dengue, has comparable rates of virus dissemination to its primary vector, Ae. aegypti, and when the longer lifespan of Ae. albopictus is considered its competence for dengue virus transmission far exceeds that of Ae. aegypti. Conclusions These results can be used to analyse the effects of temperature and other contributing factors on the expansion of dengue or its Aedes vectors. Our finding that Ae. albopictus has a greater capacity for dengue transmission than Ae. aegypti is contrary to current explanations for the comparative rarity of dengue transmission in established Ae. albopictus populations. This suggests that the limited capacity of Ae. albopictus to transmit DENV is more dependent on its ecology than vector competence. The recommendations, which we

  9. Deep sequencing reveals complex mechanisms of diapause preparation in the invasive mosquito, Aedes albopictus.

    PubMed

    Poelchau, Monica F; Reynolds, Julie A; Elsik, Christine G; Denlinger, David L; Armbruster, Peter A

    2013-05-22

    Seasonal environments present fundamental physiological challenges to a wide range of insects. Many temperate insects surmount the exigencies of winter by undergoing photoperiodic diapause, in which photoperiod provides a token cue that initiates an alternative developmental programme leading to dormancy. Pre-diapause is a crucial preparatory phase of this process, preceding developmental arrest. However, the regulatory and physiological mechanisms of diapause preparation are largely unknown. Using high-throughput gene expression profiling in the Asian tiger mosquito, Aedes albopictus, we reveal major shifts in endocrine signalling, cell proliferation, metabolism, energy production and cellular structure across pre-diapause development. While some hallmarks of diapause, such as insulin signalling and stress response, were not important at the transcriptional level, two genes, Pepck and PCNA, appear to show diapause-induced transcriptional changes across insect taxa. These processes demonstrate physiological commonalities between Ae. albopictus pre-diapause and diapause strategies across insects, and support the idea of a genetic 'toolkit' for diapause. Observations of gene expression trends from a comparative developmental perspective suggest that individual physiological processes are delayed against a background of a fixed morphological ontogeny. Our results demonstrate how deep sequencing can provide new insights into elusive molecular bases of complex ecological adaptations.

  10. Evaluation of the Human IgG Antibody Response to Aedes albopictus Saliva as a New Specific Biomarker of Exposure to Vector Bites

    PubMed Central

    Doucoure, Souleymane; Mouchet, François; Cornelie, Sylvie; DeHecq, Jean Sébastien; Rutee, Abdul Hamid; Roca, Yelin; Walter, Annie; Hervé, Jean Pierre; Misse, Dorothée; Favier, François; Gasque, Philippe; Remoue, Franck

    2012-01-01

    Background The spread of Aedes albopictus, a vector for re-emergent arbovirus diseases like chikungunya and dengue, points up the need for better control strategies and new tools to evaluate transmission risk. Human antibody (Ab) responses to mosquito salivary proteins could represent a reliable biomarker for evaluating human-vector contact and the efficacy of control programs. Methodology/Principal Findings We used ELISA tests to evaluate specific immunoglobulin G (IgG) responses to salivary gland extracts (SGE) in adults exposed to Aedes albopictus in Reunion Island. The percentage of immune responders (88%) and levels of anti-SGE IgG Abs were high in exposed individuals. At an individual level, our results indicate heterogeneity of the exposure to Aedes albopictus bites. In addition, low-level immune cross-reactivity between Aedes albopictus and Aedes aegypti SGEs was observed, mainly in the highest responders. Conclusion/Significance Ab responses to saliva could be used as an immuno-epidemiological tool for evaluating exposure to Aedes albopictus bites. Combined with entomological and epidemiological methods, a “salivary” biomarker of exposure to Aedes albopictus could enhance surveillance of its spread and the risk of arbovirus transmission, and could be used as a direct tool for the evaluation of Aedes albopictus control strategies. PMID:22363823

  11. Larval mosquito habitat utilization and community dynamics of Aedes albopictus and Aedes japonicus (Diptera: Culicidae).

    PubMed

    Bartlett-Healy, Kristen; Unlu, Isik; Obenauer, Peter; Hughes, Tony; Healy, Sean; Crepeau, Taryn; Farajollahi, Ary; Kesavaraju, Banu; Fonseca, Dina; Schoeler, George; Gaugler, Randy; Strickman, Daniel

    2012-07-01

    Aedes albopictus (Skuse) and Ae. japonicus (Theobald) are important container-inhabiting mosquitoes that transmit disease agents, outcompete native species, and continue to expand their range in the United States. Both species deposit eggs in natural and artificial containers and thrive in peridomestic environments. The goal of our study was to examine the types and characteristics of containers that are most productive for these species in the northeastern United States. In total, 306 containers were sampled in urban, suburban, and rural areas of New Jersey. Multiple biotic and abiotic factors were recorded in an attempt to identify variables associated with the productivity of each species. Based on pupal abundance and density of container types, results showed that tires, trash cans, and planter dishes were the most important containers for Ae. albopictus, while planter dishes were the most important containers for Ae. japonicus. Container color (black and gray), material (rubber), and type (tires) were correlated with species presence for Ae. albopictus and Ae. japonicus. These factors may play a role in the selection of oviposition sites by female mosquitoes or in the survival of their progeny. Differences in species composition and abundance were detected between areas classified as urban, suburban, and rural. In urban and suburban areas, Ae. albopictus was more abundant in container habitats than Ae. japonicus; however, Ae. japonicus was more abundant in rural areas, and when water temperatures were below 14 degrees C. Our results suggest many variables can influence the presence of Ae. albopictus and Ae. japonicus in container habitats in northeastern United States.

  12. Cell lines authentication and mycoplasma detection as minimun quality control of cell lines in biobanking.

    PubMed

    Corral-Vázquez, C; Aguilar-Quesada, R; Catalina, P; Lucena-Aguilar, G; Ligero, G; Miranda, B; Carrillo-Ávila, J A

    2017-06-01

    Establishment of continuous cell lines from human normal and tumor tissues is an extended and useful methodology for molecular characterization of cancer pathophysiology and drug development in research laboratories. The exchange of these cell lines between different labs is a common practice that can compromise assays reliability due to contamination with microorganism such as mycoplasma or cells from different flasks that compromise experiment reproducibility and reliability. Great proportions of cell lines are contaminated with mycoplasma and/or are replaced by cells derived for a different origin during processing or distribution process. The scientific community has underestimated this problem and thousand of research experiment has been done with cell lines that are incorrectly identified and wrong scientific conclusions have been published. Regular contamination and authentication tests are necessary in order to avoid negative consequences of widespread misidentified and contaminated cell lines. Cell banks generate, store and distribute cell lines for research, being mandatory a consistent and continuous quality program. Methods implementation for guaranteeing both, the absence of mycoplasma and authentication in the supplied cell lines, has been performed in the Andalusian Health System Biobank. Specifically, precise results were obtained using real time PCR detection for mycoplasma and 10 STRs identification by capillary electrophoresis for cell line authentication. Advantages and disadvantages of these protocols are discussed.

  13. Local environmental and meteorological conditions influencing the invasive mosquito Ae. albopictus and arbovirus transmission risk in New York City.

    PubMed

    Little, Eliza; Bajwa, Waheed; Shaman, Jeffrey

    2017-08-01

    Ae. albopictus, an invasive mosquito vector now endemic to much of the northeastern US, is a significant public health threat both as a nuisance biter and vector of disease (e.g. chikungunya virus). Here, we aim to quantify the relationships between local environmental and meteorological conditions and the abundance of Ae. albopictus mosquitoes in New York City. Using statistical modeling, we create a fine-scale spatially explicit risk map of Ae. albopictus abundance and validate the accuracy of spatiotemporal model predictions using observational data from 2016. We find that the spatial variability of annual Ae. albopictus abundance is greater than its temporal variability in New York City but that both local environmental and meteorological conditions are associated with Ae. albopictus numbers. Specifically, key land use characteristics, including open spaces, residential areas, and vacant lots, and spring and early summer meteorological conditions are associated with annual Ae. albopictus abundance. In addition, we investigate the distribution of imported chikungunya cases during 2014 and use these data to delineate areas with the highest rates of arboviral importation. We show that the spatial distribution of imported arboviral cases has been mostly discordant with mosquito production and thus, to date, has provided a check on local arboviral transmission in New York City. We do, however, find concordant areas where high Ae. albopictus abundance and chikungunya importation co-occur. Public health and vector control officials should prioritize control efforts to these areas and thus more cost effectively reduce the risk of local arboviral transmission. The methods applied here can be used to monitor and identify areas of risk for other imported vector-borne diseases.

  14. Local environmental and meteorological conditions influencing the invasive mosquito Ae. albopictus and arbovirus transmission risk in New York City

    PubMed Central

    Bajwa, Waheed; Shaman, Jeffrey

    2017-01-01

    Ae. albopictus, an invasive mosquito vector now endemic to much of the northeastern US, is a significant public health threat both as a nuisance biter and vector of disease (e.g. chikungunya virus). Here, we aim to quantify the relationships between local environmental and meteorological conditions and the abundance of Ae. albopictus mosquitoes in New York City. Using statistical modeling, we create a fine-scale spatially explicit risk map of Ae. albopictus abundance and validate the accuracy of spatiotemporal model predictions using observational data from 2016. We find that the spatial variability of annual Ae. albopictus abundance is greater than its temporal variability in New York City but that both local environmental and meteorological conditions are associated with Ae. albopictus numbers. Specifically, key land use characteristics, including open spaces, residential areas, and vacant lots, and spring and early summer meteorological conditions are associated with annual Ae. albopictus abundance. In addition, we investigate the distribution of imported chikungunya cases during 2014 and use these data to delineate areas with the highest rates of arboviral importation. We show that the spatial distribution of imported arboviral cases has been mostly discordant with mosquito production and thus, to date, has provided a check on local arboviral transmission in New York City. We do, however, find concordant areas where high Ae. albopictus abundance and chikungunya importation co-occur. Public health and vector control officials should prioritize control efforts to these areas and thus more cost effectively reduce the risk of local arboviral transmission. The methods applied here can be used to monitor and identify areas of risk for other imported vector-borne diseases. PMID:28832586

  15. Updated Reported Distribution of Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus (Diptera: Culicidae) in the United States, 1995-2016.

    PubMed

    Hahn, Micah B; Eisen, Lars; McAllister, Janet; Savage, Harry M; Mutebi, John-Paul; Eisen, Rebecca J

    2017-09-01

    Aedes (Stegomyia) aegypti (L.) and Aedes (Stegomyia) albopictus (Skuse) are potential vectors of Zika, dengue, and chikungunya viruses in the United States. A Zika virus outbreak in Florida in the summer of 2016, driven by Ae. aegypti and resulting in > 200 locally acquired cases of human illness, underscored the need for up-to-date information on the geographic distribution of Ae. aegypti and Ae. albopictus in the United States. In early 2016, we conducted a survey and literature review to compile county records for presence of Ae. aegypti and Ae. albopictus in the United States from 1995 to 2016. Surveillance for these vectors was intensified across the United States during the summer and fall of 2016. At the end of 2016, we therefore conducted a follow-up survey of mosquito control agencies, university researchers, and state and local health departments to document new collection records for Ae. aegypti and Ae. albopictus. The repeated survey at the end of the year added Ae. aegypti collection records from 38 new counties and Ae. albopictus collection records from 127 new counties, representing a 21 and 10 percent increase, respectively, in the number of counties with reported presence of these mosquitoes compared with the previous report. Moreover, through our updated survey, 40 and 183 counties, respectively, added additional years of collection records for Ae. aegypti and Ae. albopictus from 1995 to 2016. Our findings underscore the continued need for systematic surveillance of Ae. aegypti and Ae. albopictus. Published by Oxford University Press on behalf of Entomological Society of America 2017. This work is written by US Government employees and is in the public domain in the US.

  16. First mass development of Aedes albopictus (Diptera: Culicidae)-its surveillance and control in Germany.

    PubMed

    Becker, Norbert; Schön, Stefanie; Klein, Alexandra-Maria; Ferstl, Ina; Kizgin, Ali; Tannich, Egbert; Kuhn, Carola; Pluskota, Björn; Jöst, Artur

    2017-03-01

    The Asian tiger mosquito Aedes albopictus has undergone a dramatic expansion of its range in the last few decades. Since its first detection in 2007 in Germany at the motorway A5 coming from Italy via Switzerland to Germany, it has been continuously introduced by vehicles, most probably from Italy. After a hint from an alert gardener in an allotment garden area in Freiburg, Southwest Germany, in 2015, a surveillance programme was started focusing on the garden area and adjacent areas as well as most of the cemeteries as potential infestation areas. The surveillance programme confirmed a high infestation of the allotment garden. The container index (CI) exceeded almost 30% in August 2015. In lethal gravid Aedes traps (GATs) and BG-Sentinel traps, 4038 adults were caught. It could be proven that the Aedes population is more or less still spatially restricted to the allotment garden area which is adjacent to a train station where trucks from Novara, Italy, arrive loaded on trains. Outside the garden area, only a few breeding sites with developmental stages and adults were found within a radius of approximately 600 m from the highly infested garden area. It is most likely that Ae. albopictus females are constantly introduced as 'blind passengers' to Freiburg via trucks from Italy to Freiburg, Germany. After the first detection of the mass development of Ae. albopictus immediate and comprehensive control measures were initiated to reduce or even eliminate the Aedes population. Citizen awareness, especially of the gardeners, was increased by providing thorough information about the biology and control of Ae. albopictus. Beside environmental management, tablets based on Bacillus thuringiensis israelensis (Bti) were applied. The success of the control activities by the gardeners is reflected by the data gained during monthly inspection of the garden plots. The number of gardens without any container increased from 17% in July to 22% in August and 35% in September, 2015

  17. Further specimens of the Asian tiger mosquito Aedes albopictus (Diptera, Culicidae) trapped in southwest Germany.

    PubMed

    Kampen, Helge; Kronefeld, Mandy; Zielke, Dorothee; Werner, Doreen

    2013-02-01

    After two previous demonstrations of introductions of the Asian tiger mosquito, Aedes albopictus, into southern Germany in 2007 and 2011, another three specimens were trapped in the city of Freiburg in the Upper Rhine Valley. The females were caught in early September 2011 (n = 2) and mid-July 2012 (n = 1). The trap was located at a railway container station where cargo is transferred to trains from trucks predominantly coming from southern Europe where A. albopictus is widely distributed. The reported findings confirm vehicle transport of A. albopictus to be an important and probably frequent mode of importation, and suggest that more regular and intense monitoring for invasive mosquito species in the Upper Rhine Valley should be undertaken in order to detect an establishment and implement adequate control measures in good time.

  18. Seasonal patterns of oviposition and egg hatching rate of Aedes albopictus in Rome.

    PubMed

    Toma, Luciano; Severini, Francesco; Di Luca, Marco; Bella, Antonino; Romi, Roberto

    2003-03-01

    Since its introduction in Italy in 1990, Aedes albopictus has spread quickly across the northern and central regions of the country. The Italian populations of the species probably originated from temperate areas and are able to survive the cold season through the production of diapausing eggs. In 1997, Ae. albopictus was detected in Rome, where it seems to have found ideal environmental conditions for proliferating and extending its season of activity. In 2000, we carried out a study to evaluate the length of the season favorable to the species in Rome and the factors that might induce production of diapausing eggs. Adults of the species were reported to be active from February-March to December, peaking in August-September. Female Ae. albopictus produced different numbers of summer eggs throughout the study. The peak of egg hatching occurred at the 35th week of the year (end of August). Starting from the 36th week, when the photoperiod dropped below 14:10 h light:dark, a rapid decrease in summer egg production was recorded; the minimum value (17%) was reached during the 1st week of October. A new rise in egg hatching was recorded in October-November, probably because of a polymorphism in the diapause response of the population of Ae. albopictus and to the persistence of mild temperatures up to the end of November. Eggs completely ceased to hatch in mid-December.

  19. A Rainfall- and Temperature-Driven Abundance Model for Aedes albopictus Populations

    PubMed Central

    Tran, Annelise; L’Ambert, Grégory; Lacour, Guillaume; Benoît, Romain; Demarchi, Marie; Cros, Myriam; Cailly, Priscilla; Aubry-Kientz, Mélaine; Balenghien, Thomas; Ezanno, Pauline

    2013-01-01

    The mosquito Aedes (Stegomyia) albopictus (Skuse) (Diptera: Culicidae) is an invasive species which has colonized Southern Europe in the last two decades. As it is a competent vector for several arboviruses, its spread is of increasing public health concern, and there is a need for appropriate monitoring tools. In this paper, we have developed a modelling approach to predict mosquito abundance over time, and identify the main determinants of mosquito population dynamics. The model is temperature- and rainfall-driven, takes into account egg diapause during unfavourable periods, and was used to model the population dynamics of Ae. albopictus in the French Riviera since 2008. Entomological collections of egg stage from six locations in Nice conurbation were used for model validation. We performed a sensitivity analysis to identify the key parameters of the mosquito population dynamics. Results showed that the model correctly predicted entomological field data (Pearson r correlation coefficient values range from 0.73 to 0.93). The model’s main control points were related to adult’s mortality rates, the carrying capacity in pupae of the environment, and the beginning of the unfavourable period. The proposed model can be efficiently used as a tool to predict Ae. albopictus population dynamics, and to assess the efficiency of different control strategies. PMID:23624579

  20. Molluscan cells in culture: primary cell cultures and cell lines

    PubMed Central

    Yoshino, T. P.; Bickham, U.; Bayne, C. J.

    2013-01-01

    In vitro cell culture systems from molluscs have significantly contributed to our basic understanding of complex physiological processes occurring within or between tissue-specific cells, yielding information unattainable using intact animal models. In vitro cultures of neuronal cells from gastropods show how simplified cell models can inform our understanding of complex networks in intact organisms. Primary cell cultures from marine and freshwater bivalve and gastropod species are used as biomonitors for environmental contaminants, as models for gene transfer technologies, and for studies of innate immunity and neoplastic disease. Despite efforts to isolate proliferative cell lines from molluscs, the snail Biomphalaria glabrata Say, 1818 embryonic (Bge) cell line is the only existing cell line originating from any molluscan species. Taking an organ systems approach, this review summarizes efforts to establish molluscan cell cultures and describes the varied applications of primary cell cultures in research. Because of the unique status of the Bge cell line, an account is presented of the establishment of this cell line, and of how these cells have contributed to our understanding of snail host-parasite interactions. Finally, we detail the difficulties commonly encountered in efforts to establish cell lines from molluscs and discuss how these difficulties might be overcome. PMID:24198436

  1. Aedes albopictus and Aedes japonicus - two invasive mosquito species with different temperature niches in Europe.

    PubMed

    Cunze, Sarah; Koch, Lisa K; Kochmann, Judith; Klimpel, Sven

    2016-11-04

    Aedes albopictus and Ae. japonicus are two of the most widespread invasive mosquito species that have recently become established in western Europe. Both species are associated with the transmission of a number of serious diseases and are projected to continue their spread in Europe. In the present study, we modelled the habitat suitability for both species under current and future climatic conditions by means of an Ensemble forecasting approach. We additionally compared the modelled MAXENT niches of Ae. albopictus and Ae. japonicus regarding temperature and precipitation requirements. Both species were modelled to find suitable habitat conditions in distinct areas within Europe: Ae. albopictus within the Mediterranean regions in southern Europe, Ae. japonicus within the more temperate regions of central Europe. Only in few regions, suitable habitat conditions were projected to overlap for both species. Whereas Ae. albopictus is projected to be generally promoted by climate change in Europe, the area modelled to be climatically suitable for Ae. japonicus is projected to decrease under climate change. This projection of range reduction under climate change relies on the assumption that Ae. japonicus is not able to adapt to warmer climatic conditions. The modelled MAXENT temperature niches of Ae. japonicus were found to be narrower with an optimum at lower temperatures compared to the niches of Ae. albopictus. Species distribution models identifying areas with high habitat suitability can help improving monitoring programmes for invasive species currently in place. However, as mosquito species are known to be able to adapt to new environmental conditions within the invasion range quickly, niche evolution of invasive mosquito species should be closely followed upon in future studies.

  2. Aedes albopictus (Diptera: Culicidae) as a potential vector of endemic and exotic arboviruses in Australia.

    PubMed

    Nicholson, J; Ritchie, S A; van den Hurk, A F

    2014-05-01

    In 2005, established populations of Aedes albopictus (Skuse) were discovered in the Torres Strait, the region that separates Papua New Guinea from northern Australia. This increased the potential for this species to be introduced to mainland Australia. Because it is an arbovirus vector elsewhere, we undertook laboratory-based infection and transmission experiments to determine the potential for Ae. albopictus from the Torres Strait to become infected with and transmit the four major Australian endemic arboviruses--Murray Valley encephalitis virus, West Nile virus Kunjin strain (WNV(KUN)), Ross River virus (RRV), and Barmah Forest virus--as well as the exotic Japanese encephalitis virus. Ae. albopictus is susceptible to infection with all viruses, with infection rates ranging between 8% for WNV(KUN) and 71% for RRV. Transmission rates of approximately 25% were observed for RRV and Barmah Forest virus, but these were < 17% for Murray Valley encephalitis virus, WNV(KUN), and Japanese encephalitis virus. Given its relative vector competence for alphaviruses, we also examined the replication kinetics and extrinsic incubation periods required for transmission of RRV and chikungunya virus. Despite lower body titers, more mosquitoes reared and maintained at 28 degrees C became infected with and transmitted the virus than those reared and maintained at 22 degrees C. The minimum time between Ae. albopictus consuming an infected bloodmeal and transmitting chikungunya virus was 2 d at 28 degrees C and 4 d at 22 degrees C, and for RRV, it was 4 d, irrespective of the temperature. Given its opportunistic feeding habits and aggressive biting behavior, the establishment of Ae. albopictus on the Australian mainland could have a considerable impact on alphavirus transmission.

  3. Ascogregarina taiwanensis infection in Aedes aegypti and Aedes albopictus in Santa Catarina, South Brazil.

    PubMed

    Prophiro, Josiane Somariva; Pereira, Thiago Nunes; Oliveira, Joice Guilherme de; Dandolini, Guilherme Werner; Silva, Mario Antonio Navarro da; Silva, Onilda Santos da

    2017-01-01

    This study registers Ascogregarina spp. infection in field populations of Aedes aegypti and Aedes albopictus in a subtropical region of Brazil. Mosquito larvae collected in tires placed in four municipalities of Santa Catarina were identified morphologically and assessed for Ascogregarina sp. infection using morphological and molecular methods. Both mosquito species harbored Ascogregarina taiwanensis, whose genomic DNA was confirmed in both the Aedes species by PCR. DNA sequences were deposited in GenBank. Conclusion: Both Ae. albopictus e Ae. aegypti harbor Ascogregarina sp.

  4. Population genetics of the Asian tiger mosquito Aedes albopictus, an invasive vector of human diseases

    PubMed Central

    Goubert, C; Minard, G; Vieira, C; Boulesteix, M

    2016-01-01

    The Asian tiger mosquito Aedes albopictus is currently one of the most threatening invasive species in the world. Native to Southeast Asia, the species has spread throughout the world in the past 30 years and is now present in every continent but Antarctica. Because it was the main vector of recent Dengue and Chikungunya outbreaks, and because of its competency for numerous other viruses and pathogens such as the Zika virus, A. albopictus stands out as a model species for invasive diseases vector studies. A synthesis of the current knowledge about the genetic diversity of A. albopictus is needed, knowing the interplays between the vector, the pathogens, the environment and their epidemiological consequences. Such resources are also valuable for assessing the role of genetic diversity in the invasive success. We review here the large but sometimes dispersed literature about the population genetics of A. albopictus. We first debate about the experimental design of these studies and present an up-to-date assessment of the available molecular markers. We then summarize the main genetic characteristics of natural populations and synthesize the available data regarding the worldwide structuring of the vector. Finally, we pinpoint the gaps that remain to be addressed and suggest possible research directions. PMID:27273325

  5. Cell Line Data Base: structure and recent improvements towards molecular authentication of human cell lines

    PubMed Central

    Romano, Paolo; Manniello, Assunta; Aresu, Ottavia; Armento, Massimiliano; Cesaro, Michela; Parodi, Barbara

    2009-01-01

    The Cell Line Data Base (CLDB) is a well-known reference information source on human and animal cell lines including information on more than 6000 cell lines. Main biological features are coded according to controlled vocabularies derived from international lists and taxonomies. HyperCLDB (http://bioinformatics.istge.it/hypercldb/) is a hypertext version of CLDB that improves data accessibility by also allowing information retrieval through web spiders. Access to HyperCLDB is provided through indexes of biological characteristics and navigation in the hypertext is granted by many internal links. HyperCLDB also includes links to external resources. Recently, an interest was raised for a reference nomenclature for cell lines and CLDB was seen as an authoritative system. Furthermore, to overcome the cell line misidentification problem, molecular authentication methods, such as fingerprinting, single-locus short tandem repeat (STR) profile and single nucleotide polymorphisms validation, were proposed. Since this data is distributed, a reference portal on authentication of human cell lines is needed. We present here the architecture and contents of CLDB, its recent enhancements and perspectives. We also present a new related database, the Cell Line Integrated Molecular Authentication (CLIMA) database (http://bioinformatics.istge.it/clima/), that allows to link authentication data to actual cell lines. PMID:18927105

  6. Cell Line Data Base: structure and recent improvements towards molecular authentication of human cell lines.

    PubMed

    Romano, Paolo; Manniello, Assunta; Aresu, Ottavia; Armento, Massimiliano; Cesaro, Michela; Parodi, Barbara

    2009-01-01

    The Cell Line Data Base (CLDB) is a well-known reference information source on human and animal cell lines including information on more than 6000 cell lines. Main biological features are coded according to controlled vocabularies derived from international lists and taxonomies. HyperCLDB (http://bioinformatics.istge.it/hypercldb/) is a hypertext version of CLDB that improves data accessibility by also allowing information retrieval through web spiders. Access to HyperCLDB is provided through indexes of biological characteristics and navigation in the hypertext is granted by many internal links. HyperCLDB also includes links to external resources. Recently, an interest was raised for a reference nomenclature for cell lines and CLDB was seen as an authoritative system. Furthermore, to overcome the cell line misidentification problem, molecular authentication methods, such as fingerprinting, single-locus short tandem repeat (STR) profile and single nucleotide polymorphisms validation, were proposed. Since this data is distributed, a reference portal on authentication of human cell lines is needed. We present here the architecture and contents of CLDB, its recent enhancements and perspectives. We also present a new related database, the Cell Line Integrated Molecular Authentication (CLIMA) database (http://bioinformatics.istge.it/clima/), that allows to link authentication data to actual cell lines.

  7. Bioecology and vectorial capacity of Aedes albopictus (Diptera: Culicidae) in Macao, China, in relation to dengue virus transmission.

    PubMed

    Almeida, A Paulo G; Baptista, Susana S S G; Sousa, Carla A G C C; Novo, M Teresa L M; Ramos, Helena C; Panella, Nicholas A; Godsey, Marvin; Simões, M João; Anselmo, M Luisa; Komar, Nicholas; Mitchell, Carl J; Ribeiro, Henrique

    2005-05-01

    Until 2001, the Chinese Territory of Macao had not registered any autochthonous dengue cases, despite the abundance of Aedes albopictus (Skuse), a known vector. This work describes a bioecological characterization of the local Ae. albopictus adult population, with the purpose of estimating the receptivity of Macao to dengue introduction. In the wet seasons of 1997 and 1998 and the dry season of 1998, Ae. albopictus was the most abundant human-biting mosquito. Daily biting rates of 314 mosquitoes per person were recorded in the wet season with a reduction to 94 in the dry season. Ae. albopictus was mainly exophagic and exophilic and had a human blood index of 44%. The parity rate of field-collected mosquitoes was 57%. Daily survival rate ranged from 91 to 97%. Estimates of vectorial capacity ranged from 144 to 880, depending on what parameter values were used. These estimates indicated a great receptivity for the introduction of dengue viruses, as the 2001 outbreak came to prove.

  8. Molecular and Functional Characterization of Odorant-Binding Protein Genes in an Invasive Vector Mosquito, Aedes albopictus

    PubMed Central

    Deng, Yuhua; Yan, Hui; Gu, Jinbao; Xu, Jiabao; Wu, Kun; Tu, Zhijian; James, Anthony A.; Chen, Xiaoguang

    2013-01-01

    Aedes albopictus is a major vector of dengue and Chikungunya viruses. Olfaction plays a vital role in guiding mosquito behaviors and contributes to their ability to transmit pathogens. Odorant-binding proteins (OBPs) are abundant in insect olfactory tissues and involved in the first step of odorant reception. While comprehensive descriptions are available of OBPs from Aedes aegypti, Culex quinquefasciatus and Anopheles gambiae, only a few genes from Ae. albopictus have been reported. In this study, twenty-one putative AalbOBP genes were cloned using their homologues in Ae. aegypti to query an Ae. albopictus partial genome sequence. Two antenna-specific OBPs, AalbOBP37 and AalbOBP39, display a remarkable similarity in their overall folding and binding pockets, according to molecular modeling. Binding affinity assays indicated that AalbOBP37 and AalbOBP39 had overlapping ligand affinities and are affected in different pH condition. Electroantennagrams (EAG) and behavioral tests show that these two genes were involved in olfactory reception. An improved understanding of the Ae. albopictus OBPs is expected to contribute to the development of more efficient and environmentally-friendly mosquito control strategies. PMID:23935894

  9. Radiation sensitivity of Merkel cell carcinoma cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Leonard, J.H.; Ramsay, J.R.; Birrell, G.W.

    1995-07-30

    Merkel cell carcinoma (MCC), being a small cell carcinoma, would be expected to be sensitive to radiation. Clinical analysis of patients at our center, especially those with macroscopic disease, would suggest the response is quite variable. We have recently established a number of MCC cell lines from patients prior to radiotherapy, and for the first time are in a position to determine their sensitivity under controlled conditions. Some of the MCC lines grew as suspension cultures and could not be single cell cloned; therefore, it was not possible to use clonogenic survival for all cell lines. A tetrazolium based (MTT)more » assay was used for these lines, to estimate cell growth after {gamma} irradiation. Control experiments were conducted on lymphoblastoid cell lines (LCL) and the adherent MCC line, MCC13, to demonstrate that the two assays were comparable under the conditions used. We have examined cell lines from MCC, small cell lung cancer (SCLC), malignant melanomas, Epstein Barr virus (EBV) transformed lymphocytes (LCL), and skin fibroblasts for their sensitivity to {gamma} irradiation using both clonogenic cell survival and MTT assays. The results show that the tumor cell lines have a range of sensitivities, with melanoma being more resistant (surviving fraction at 2 Gy (SF2) 0.57 and 0.56) than the small cell carcinoma lines, MCC (SF2 range 0.21-0.45, mean SF2 0.30, n = 8) and SCLC (SF2 0.31). Fibroblasts were the most sensitive (SF2 0.13-0.20, mean 0.16, n = 5). The MTT assay, when compared to clonogenic assay for the MCC13 adherent line and the LCL, gave comparable results under the conditions used. Both assays gave a range of SF2 values for the MCC cell lines, suggesting that these cancers would give a heterogeneous response in vivo. The results with the two derivative clones of MCC14 (SF2 for MCC14/1 0.38, MCC14/2 0.45) would further suggest that some of them may develop resistance during clonogenic evolution. 25 refs., 3 figs., 1 tab.« less

  10. Truck-mounted Area-wide Application of Pyriproxyfen Targeting Aedes aegypti and Aedes albopictus in Northeast Florida

    DTIC Science & Technology

    2014-12-01

    the time and the lower rate of pyriproxyfen applied. Within 2 wk following Spray 2, however, Ae. albopictus collections from the treatment plot...districts/public health agencies. Pyriproxyfen, a juvenile hormone mimic, func- tions as an insect growth regulator (IGR) preventing normal development of...the lower rate of pyriproxyfen applied. Within 2 wk following Spray 2, however, Ae. albopictus collections from the treatment plot averaged

  11. Cold acclimation and overwintering of female Aedes albopictus in Roma.

    PubMed

    Romi, Roberto; Severini, Francesco; Toma, Luciano

    2006-03-01

    Eight years after the introduction and establishment of Aedes albopictus in Roma, females of the species extended their trophic activity to the coldest months of the year. Winter monitoring carried out from December 2003 to March 2004 in the urban area of the capital city of Italy recorded a weekly rate of positive ovitraps constantly around 30%, for the entire period of surveillance (14 wk). Eggs from ovitraps did not hatch when stimulated in laboratory with a wet and dry procedure. The hypothesis that long-lived female Ae. albopictus, belonging to the last seasonal generation, could have continued their trophic activity for 3 months, or that 1 or more cycles of reproduction have occurred in peculiar breeding sites, where a favorable microclimate may have allowed the development of at least 1 larval generation, are discussed and compared with the existing literature.

  12. Effects of tire leachate on the invasive mosquito Aedes albopictus and the native congener Aedes triseriatus.

    PubMed

    Villena, Oswaldo C; Terry, Ivana; Iwata, Kayoko; Landa, Edward R; LaDeau, Shannon L; Leisnham, Paul T

    2017-01-01

    Discarded vehicle tire casings are an important artificial habitat for the developmental stages of numerous vector mosquitoes. Discarded vehicle tires degrade under ultraviolet light and leach numerous soluble metals (e.g., barium, cadmium, zinc) and organic substances (e.g., benzothiazole and its derivatives [BZTs], polyaromatic hydrocarbons [PAHs]) that could affect mosquito larvae that inhabit the tire casing. This study examined the relationship between soluble zinc, a common marker of tire leachate, on mosquito densities in tire habitats in the field, and tested the effects of tire leachate on the survival and development of newly hatched Aedes albopictus and Aedes triseriatus larvae in a controlled laboratory dose-response experiment. In the field, zinc concentrations were as high as 7.26 mg/L in a single tire and averaged as high as 2.39 (SE ± 1.17) mg/L among tires at a single site. Aedes albopictus (37/42 tires, 81.1%) and A. triseriatus (23/42, 54.8%) were the most widespread mosquito species, co-occurred in over half (22/42, 52.4%) of all tires, and A. triseriatus was only collected without A. albopictus in one tire. Aedes triseriatus was more strongly negatively associated with zinc concentration than A. albopictus, and another common mosquito, C. pipiens, which was found in 17 tires. In the laboratory experiment, A. albopictus per capita rate of population change ( λ ') was over 1.0, indicating positive population growth, from 0-8.9 mg/L zinc concentration (0-10,000 mg/L tire leachate), but steeply declined to zero from 44.50-89.00 mg/L zinc (50,000-100,000 mg/L tire leachate). In contrast, A. triseriatus λ ' declined at the lower concentration of 0.05 mg/L zinc (100 mg/L tire leachate), and was zero at 0.45, 8.90, 44.50, and 89.00 mg/L zinc (500, 10,000, 50,000 and 100,000 mg/L tire leachate). These results indicate that tire leachate can have severe negative effects on populations of container-utilizing mosquitoes at concentrations

  13. Effects of tire leachate on the invasive mosquito Aedes albopictus and the native congener Aedes triseriatus

    PubMed Central

    Villena, Oswaldo C.; Terry, Ivana; Iwata, Kayoko; Landa, Edward R.; LaDeau, Shannon L.

    2017-01-01

    Discarded vehicle tire casings are an important artificial habitat for the developmental stages of numerous vector mosquitoes. Discarded vehicle tires degrade under ultraviolet light and leach numerous soluble metals (e.g., barium, cadmium, zinc) and organic substances (e.g., benzothiazole and its derivatives [BZTs], polyaromatic hydrocarbons [PAHs]) that could affect mosquito larvae that inhabit the tire casing. This study examined the relationship between soluble zinc, a common marker of tire leachate, on mosquito densities in tire habitats in the field, and tested the effects of tire leachate on the survival and development of newly hatched Aedes albopictus and Aedes triseriatus larvae in a controlled laboratory dose-response experiment. In the field, zinc concentrations were as high as 7.26 mg/L in a single tire and averaged as high as 2.39 (SE ± 1.17) mg/L among tires at a single site. Aedes albopictus (37/42 tires, 81.1%) and A. triseriatus (23/42, 54.8%) were the most widespread mosquito species, co-occurred in over half (22/42, 52.4%) of all tires, and A. triseriatus was only collected without A. albopictus in one tire. Aedes triseriatus was more strongly negatively associated with zinc concentration than A. albopictus, and another common mosquito, C. pipiens, which was found in 17 tires. In the laboratory experiment, A. albopictus per capita rate of population change (λ′) was over 1.0, indicating positive population growth, from 0–8.9 mg/L zinc concentration (0–10,000 mg/L tire leachate), but steeply declined to zero from 44.50–89.00 mg/L zinc (50,000–100,000 mg/L tire leachate). In contrast, A. triseriatus λ′ declined at the lower concentration of 0.05 mg/L zinc (100 mg/L tire leachate), and was zero at 0.45, 8.90, 44.50, and 89.00 mg/L zinc (500, 10,000, 50,000 and 100,000 mg/L tire leachate). These results indicate that tire leachate can have severe negative effects on populations of container-utilizing mosquitoes at

  14. BHD Tumor Cell Line and Renal Cell Carcinoma Line | NCI Technology Transfer Center | TTC

    Cancer.gov

    Scientists at the National Cancer Institute  have developed a novel renal cell carcinoma (RCC) cell line designated UOK257, which was derived from the surgical kidney tissue of a patient with hereditary Birt-Hogg-Dube''''(BHD) syndrome and companion cell line UOK257-2 in which FLCN expression has been restored by lentivirus infection. The NCI Urologic Oncology Branch seeks parties interested in licensing or collaborative research to co-develop, evaluate, or commercialize kidney cancer tumor cell lines.

  15. Biogeography of the two major arbovirus mosquito vectors, Aedes aegypti and Aedes albopictus (Diptera, Culicidae), in Madagascar.

    PubMed

    Raharimalala, Fara Nantenaina; Ravaomanarivo, Lala Harivelo; Ravelonandro, Pierre; Rafarasoa, Lala Sahondra; Zouache, Karima; Tran-Van, Van; Mousson, Laurence; Failloux, Anna-Bella; Hellard, Eléonore; Moro, Claire Valiente; Ralisoa, Bakoly Olga; Mavingui, Patrick

    2012-03-20

    In the past ten years, the Indian Ocean region has been the theatre of severe epidemics of chikungunya and dengue. These outbreaks coincided with a high increase in populations of Aedes albopictus that outcompete its sister taxon Aedes aegypti in most islands sampled. The objective of this work was to update the entomological survey of the two Aedes species in the island of Madagascar which has to face these arboviroses. The sampling of Aedes mosquitoes was conducted during two years, from October 2007 to October 2009, in fifteen localities from eight regions of contrasting climates. Captured adults were identified immediately whereas immature stages were bred until adult stage for determination. Phylogenetic analysis was performed using two mtDNA genes, COI and ND5 and trees were constructed by the maximum likelihood (ML) method with the gene time reversible (GTR) model. Experimental infections with the chikungunya virus strain 06.21 at a titer of 107.5 pfu/mL were performed to evaluate the vector competence of field-collected mosquitoes. Disseminated infection rates were measured fourteen days after infection by immunofluorescence assay performed on head squashes. The species Aedes aegypti was detected in only six sites in native forests and natural reserves. In contrast, the species Aedes albopictus was found in 13 out of the 15 sites sampled. Breeding sites were mostly found in man-made environments such as discarded containers, used tires, abandoned buckets, coconuts, and bamboo cuts. Linear regression models showed that the abundance of Ae. albopictus was significantly influenced by the sampling region (F = 62.00, p < 2.2 × 10(-16)) and period (F = 36.22, p = 2.548 × 10(-13)), that are associated with ecological and climate variations. Phylogenetic analysis of the invasive Ae. albopictus distinguished haplotypes from South Asia and South America from those of Madagascar, but the markers used were not discriminant enough to discern Malagasy populations. The

  16. First report of the Phe1534Cys kdr mutation in natural populations of Aedes albopictus from Brazil.

    PubMed

    Aguirre-Obando, Oscar Alexander; Martins, Ademir Jesus; Navarro-Silva, Mário Antônio

    2017-03-27

    Knockdown resistance (kdr), caused by alterations in the voltage-gated sodium channel (Na V ), is one of the mechanisms responsible for pyrethroid (PY) resistance. In the Asian tiger mosquito, Aedes albopictus, at least four different mutations were described in the IIIS6 Na V segment in populations from Asia, North America and Europe. In contrast, in Aedes aegypti at least 12 non-synonymous mutations have been reported at nine different codons, mostly in the IIS6 and IIIS6 Na V segments. The Phe1534Cys kdr mutation in the IIIS6 Na V segment is the most prevalent in populations of Ae. aegypti worldwide, also found in Ae. albopictus from Singapore. Herein, we investigated the DNA diversity corresponding to the IIS6 and IIIS6 Na V segments in natural populations of Ae. albopictus from Brazil. DNA from eight Brazilian Ae. albopictus natural populations were individually extracted and pooled by states of origin, amplified, cloned and sequenced for the corresponding IIS6 and IIIS6 Na V segments. Additionally, samples from each location were individually genotyped by an allelic specific PCR (AS-PCR) approach to obtain the genotypic and allelic frequencies for the 1534 Na V site. No non-synonymous substitutions were observed in the IIS6 sequences. However, the Phe1534Cys kdr mutation was evidenced in the Ae. albopictus Na V IIIS6 segment sequences from Paraná (PR) and Rondônia (RO) states, but not from Mato Grosso (MT) state. The 1534Cys kdr allele varied from 3% (Marilena/PR and Porto Velho/RO) to 10% (Foz do Iguaçu/PR). To our knowledge, this paper reports the first occurrence and provides distribution data of a possible kdr mutation in Ae. albopictus in South America. The emergence of a likely kdr mutation in Ae. albopitus natural populations is a signal of alert for vector control measures since PY are the most popular insecticides adopted by residents. Additionally, once the kdr allele is present, its frequency tends to increase faster under exposition to those

  17. Comparison of Dengue Virus Type 2-Specific Small RNAs from RNA Interference-Competent and –Incompetent Mosquito Cells

    PubMed Central

    Scott, Jaclyn C.; Brackney, Doug E.; Campbell, Corey L.; Bondu-Hawkins, Virginie; Hjelle, Brian; Ebel, Greg D.; Olson, Ken E.; Blair, Carol D.

    2010-01-01

    The exogenous RNA interference (RNAi) pathway is an important antiviral defense against arboviruses in mosquitoes, and virus-specific small interfering (si)RNAs are key components of this pathway. Understanding the biogenesis of siRNAs in mosquitoes could have important ramifications in using RNAi to control arbovirus transmission. Using deep sequencing technology, we characterized dengue virus type 2 (DENV2)-specific small RNAs produced during infection of Aedes aegypti mosquitoes and A. aegypti Aag2 cell cultures and compared them to those produced in the C6/36 Aedes albopictus cell line. We show that the size and mixed polarity of virus-specific small RNAs from DENV-infected A. aegypti cells indicate that they are products of Dicer-2 (Dcr2) cleavage of long dsRNA, whereas C6/36 cells generate DENV2-specific small RNAs that are longer and predominantly positive polarity, suggesting that they originate from a different small RNA pathway. Examination of virus-specific small RNAs after infection of the two mosquito cell lines with the insect-only flavivirus cell fusing agent virus (CFAV) corroborated these findings. An in vitro assay also showed that Aag2 A. aegypti cells are capable of siRNA production, while C6/36 A. albopictus cells exhibit inefficient Dcr2 cleavage of long dsRNA. Defective expression or function of Dcr2, the key initiator of the RNAi pathway, might explain the comparatively robust growth of arthropod-borne viruses in the C6/36 cell line, which has been used frequently as a surrogate for studying molecular interactions between arboviruses and cells of their mosquito hosts. PMID:21049014

  18. Modeling the habitat suitability for the arbovirus vector Aedes albopictus (Diptera: Culicidae) in Germany.

    PubMed

    Koch, Lisa K; Cunze, Sarah; Werblow, Antje; Kochmann, Judith; Dörge, Dorian D; Mehlhorn, Heinz; Klimpel, Sven

    2016-03-01

    Climatic changes raise the risk of re-emergence of arthropod-borne virus outbreaks globally. These viruses are transmitted by arthropod vectors, often mosquitoes. Due to increasing worldwide trade and tourism, these vector species are often accidentally introduced into many countries beyond their former distribution range. Aedes albopictus, a well-known disease vector, was detected for the first time in Germany in 2007, but seems to have failed establishment until today. However, the species is known to occur in other temperate regions and a risk for establishment in Germany remains, especially in the face of predicted climate change. Thus, the goal of the study was to estimate the potential distribution of Ae. albopictus in Germany. We used ecological niche modeling in order to estimate the potential habitat suitability for this species under current and projected future climatic conditions. According to our model, there are already two areas in western and southern Germany that appear suitable for Ae. albopictus under current climatic conditions. One of these areas lies in Baden-Wuerttemberg, the other in North-Rhine Westphalia in the Ruhr region. Furthermore, projections under future climatic conditions show an increase of the modeled habitat suitability throughout Germany. Ae. albopictus is supposed to be better acclimated to colder temperatures than other tropical vectors and thus, might become, triggered by climate change, a serious threat to public health in Germany. Our modeling results can help optimizing the design of monitoring programs currently in place in Germany.

  19. The “Auto-Dissemination” Approach: A Novel Concept to Fight Aedes albopictus in Urban Areas

    PubMed Central

    Caputo, Beniamino; Ienco, Annamaria; Cianci, Daniela; Pombi, Marco; Petrarca, Vincenzo; Baseggio, Alberto; Devine, Gregor J.; della Torre, Alessandra

    2012-01-01

    Background The main constraint to the fight against container-breeding mosquito vectors of human arboviruses is the difficulty in targeting the multiplicity of larval sources, mostly represented by small man-made water containers. The aim of this work is to assess the feasibility of the “auto-dissemination” approach, already tested for Aedes aegypti, as a possible alternative to traditional, inefficient control tools, against Ae. albopictus in urban areas. The approach is based on the possibility that wild adult females, exposed to artificial resting sites contaminated with pyriproxyfen, can disseminate this juvenile hormone analogue to larval habitats, thus interfering with adult emergence. Methodology We carried out four field experiments in two areas of Rome that are typically highly infested with Ae. albopictus, i.e. the main cemetery and a small green area within a highly urbanised neighbourhood. In each area we used 10 pyriproxyfen “dissemination” stations, 10 “sentinel” sites and 10 covered, control sites. The sentinel and control sites each contained 25 Ae. albopictus larvae. These were monitored for development and adult emergence. Principal Findings When a 5% pyriproxyfen powder was used to contaminate the dissemination sites, we observed significantly higher mortality at the pupal stage in the sentinel sites (50–70%) than in the controls (<2%), showing that pyriproxyfen was transferred by mosquitoes into sentinel sites and that it had a lethal effect. Conclusions The results support the potential feasibility of the auto-dissemination approach to control Ae. albopictus in urban areas. Further studies will be carried out to optimize the method and provide an effective tool to reduce the biting nuisance caused by this aggressive species and the transmission risk of diseases such as Dengue and Chikungunya. These arboviruses pose an increasing threat in Europe as Ae. albopictus expands its range. PMID:22953015

  20. Predicting the Start of the Aedes albopictus (Diptera: Culicidae) Female Adult Biting Season Using the Spring Temperature in Japan

    PubMed Central

    Higa, Yukiko; Muto, Atsushi; Hirabayashi, Kimio; Yoshida, Masahiro; Sato, Takashi; Nihei, Naoko; Sawabe, Kyoko; Kobayashi, Mutsuo

    2017-01-01

    Abstract Aedes albopictus (Skuse) (Diptera: Culicidae) is distributed widely and is common in much of Japan. In Japan, female adults begin to bite in between April and June, except in the southern subtropics where the mosquito has no dormant period. It is difficult to estimate the first Ae. albopictus biting day because it varies annually depending on the location. Over several years, we surveyed the mosquitoes at different locations that covered a range of warmer to cooler areas of Japan. We found an association between the timing of first biting day by Ae. albopictus and spring temperature. In spring months, the strongest correlation was found with mean April temperatures, followed by March. Based on these data, it may, therefore, be possible to apply a simple formula to predict the timing of the first biting day at various geographical locations in Japan. Forecasting maps were created using a simple prediction formula. We found that the first biting day for Ae. albopictus changed depending on early spring temperatures for each year. There was an approximate 20-d difference in first biting day between years with warmer and cooler springs. This prediction model will provide useful insight for planning and practice of Ae. albopictus control programs, targeting larvae and adults, in temperate regions globally. PMID:28968909

  1. Authentication of M14 melanoma cell line proves misidentification of MDA‐MB‐435 breast cancer cell line

    PubMed Central

    Korch, Christopher; Hall, Erin M.; Dirks, Wilhelm G.; Ewing, Margaret; Faries, Mark; Varella‐Garcia, Marileila; Robinson, Steven; Storts, Douglas; Turner, Jacqueline A.; Wang, Ying; Burnett, Edward C.; Healy, Lyn; Kniss, Douglas; Neve, Richard M.; Nims, Raymond W.; Reid, Yvonne A.; Robinson, William A.

    2017-01-01

    A variety of analytical approaches have indicated that melanoma cell line UCLA‐SO‐M14 (M14) and breast carcinoma cell line MDA‐MB‐435 originate from a common donor. This indicates that at some point in the past, one of these cell lines became misidentified, meaning that it ceased to correspond to the reported donor and instead became falsely identified (through cross‐contamination or other means) as a cell line from a different donor. Initial studies concluded that MDA‐MB‐435 was the misidentified cell line and M14 was the authentic cell line, although contradictory evidence has been published, resulting in further confusion. To address this question, we obtained early samples of the melanoma cell line (M14), a lymphoblastoid cell line from the same donor (ML14), and donor serum preserved at the originator's institution. M14 samples were cryopreserved in December 1975, before MDA‐MB‐435 cells were established in culture. Through a series of molecular characterizations, including short tandem repeat (STR) profiling and cytogenetic analysis, we demonstrated that later samples of M14 and MDA‐MB‐435 correspond to samples of M14 frozen in 1975, to the lymphoblastoid cell line ML14, and to the melanoma donor's STR profile, sex and blood type. This work demonstrates conclusively that M14 is the authentic cell line and MDA‐MB‐435 is misidentified. With clear provenance information and authentication testing of early samples, it is possible to resolve debates regarding the origins of problematic cell lines that are widely used in cancer research. PMID:28940260

  2. A new tent trap for monitoring the daily activity of Aedes aegypti and Aedes albopictus.

    PubMed

    Casas Martínez, Mauricio; Orozco Bonilla, Arnoldo; Muñoz Reyes, Miguel; Ulloa García, Armando; Bond, J Guillermo; Valle Mora, Javier; Weber, Manuel; Rojas, Julio C

    2013-12-01

    In this study, we designed a new tent trap; the BioDiVector (BDV) tent trap, consisting of two rectangular tents that use human bait without endangering the technical personnel. The daily activity pattern of Aedes aegypti and Aedes albopictus in intra, peri, and extradomiciliary sites was studied in an endemic area of dengue in southern Mexico by using the BDV tent trap. Totals of 3,128 individuals of Ae. aegypti and 833 Ae. albopictus were captured. More Ae. aegypti males than females were caught, while the opposite was true with Ae. albopictus. The activity of both mosquito species was affected by the interaction between the collection site and time of day. In general, more individuals of both mosquito species were captured at the extradomicillary sites than at the peri and intradomicillary sites. Mosquitoes showed two peaks of activity, one in the morning and the other in the afternoon, but in general this only occurred at the extradomicillary sites, whereas no peak of activity was observed at the intra and peridomicillary sites. Overall, Ae. aegypti had a higher indirect biting rate than Ae. albopictus. Finally, due to its efficiency, simplicity, and low cost, we suggest the use of this innovative tool for entomological surveillance, bionomics and vector incrimination studies in geographical areas where dengue and other arboviruses are present. © 2013 The Society for Vector Ecology.

  3. Survivorship of adult Aedes albopictus (Diptera: Culicidae) feeding on indoor ornamental plants with no inflorescence.

    PubMed

    Qualls, Whitney A; Xue, Rui De; Beier, John C; Müller, Günter C

    2013-06-01

    The international trade of lucky bamboo (Dracaena sanderiana [Asparagaceae]) is responsible for certain introductions of the exotic species Aedes albopictus (Skuse) in California and the Netherlands. Understanding the association of this species with lucky bamboo and other ornamental plants is important from a public health standpoint. The aim of this study was to investigate the importance of indoor ornamental plants as sugar sources for adult A. albopictus. If exposed to D. sanderiana, bromeliad (Guzmania spp. hybrid [Bromeliaceae]), Moses-in-the-cradle (Rhoeo spathacea [Commelinaceae]), 10 % sucrose solution, and a negative water control as the only nutrient source, adult female A. albopictus mean survival time was 12, 7, 6, 15, and 4 days, respectively. Mean survival times for adult males were not significantly different (P > 0.05) from the females and were 10, 7, 6, 14, and 3 days, respectively. Combined male and female survival times were not significantly different on lucky bamboo compared to survival times on a 10 % sucrose control. Based on our findings, A. albopictus can readily survive long enough to complete a gonotrophic cycle and potentially complete the extrinsic incubation period for many arboviruses when only provided access to lucky bamboo plants or possibly other common ornamentals. Vector control professionals should be aware of potential in-home infestations and public health concerns associated with mosquito breeding and plant tissue feeding on ornamental plants.

  4. Thyroid cell lines in research on goitrogenesis.

    PubMed

    Gerber, H; Peter, H J; Asmis, L; Studer, H

    1991-12-01

    Thyroid cell lines have contributed a lot to the understanding of goitrogenesis. The cell lines mostly used in thyroid research are briefly discussed, namely the rat thyroid cell lines FRTL and FRTL-5, the porcine thyroid cell lines PORTHOS and ARTHOS, The sheep thyroid cell lines OVNIS 5H and 6H, the cat thyroid cell lines PETCAT 1 to 4 and ROMCAT, and the human thyroid cell lines FTC-133 and HTh 74. Chinese hamster ovary (CHO) cells and COS-7 cells, stably transfected with TSH receptor cDNA and expressing a functional TSH receptor, are discussed as examples for non-thyroidal cells, transfected with thyroid genes.

  5. Role of Bunyamwera Orthobunyavirus NSs protein in infection of mosquito cells.

    PubMed

    Szemiel, Agnieszka M; Failloux, Anna-Bella; Elliott, Richard M

    2012-01-01

    Bunyamwera orthobunyavirus is both the prototype and study model of the Bunyaviridae family. The viral NSs protein seems to contribute to the different outcomes of infection in mammalian and mosquito cell lines. However, only limited information is available on the growth of Bunyamwera virus in cultured mosquito cells other than the Aedes albopictus C6/36 line. To determine potential functions of the NSs protein in mosquito cells, replication of wild-type virus and a recombinant NSs deletion mutant was compared in Ae. albopictus C6/36, C7-10 and U4.4 cells, and in Ae. aegypti Ae cells by monitoring N protein production and virus yields at various times post infection. Both viruses established persistent infections, with the exception of NSs deletion mutant in U4.4 cells. The NSs protein was nonessential for growth in C6/36 and C7-10 cells, but was important for productive replication in U4.4 and Ae cells. Fluorescence microscopy studies using recombinant viruses expressing green fluorescent protein allowed observation of three stages of infection, early, acute and late, during which infected cells underwent morphological changes. In the absence of NSs, these changes were less pronounced. An RNAi response efficiently reduced virus replication in U4.4 cells transfected with virus specific dsRNA, but not in C6/36 or C7/10 cells. Lastly, Ae. aegypti mosquitoes were exposed to blood-meal containing either wild-type or NSs deletion virus, and at various times post-feeding, infection and disseminated infection rates were measured. Compared to wild-type virus, infection rates by the mutant virus were lower and more variable. If the NSs deletion virus was able to establish infection, it was detected in salivary glands at 6 days post-infection, 3 days later than wild-type virus. Bunyamwera virus NSs is required for efficient replication in certain mosquito cell lines and in Ae. aegypti mosquitoes.

  6. Chemical composition, larvicidal action, and adult repellency of Thymus magnus against Aedes albopictus.

    PubMed

    Park, Young-Uk; Koo, Hyun-Na; Kim, Gil-Hah

    2012-09-01

    Thymus magnus, an endemic species, is found in the Republic of Korea. The volatile compounds extracted by SPME from T. magnus were investigated for their chemical composition and electrophysiological response against the Asian tiger mosquito, Aedes albopictus. The volatile compounds of T. magnus as determined by gas chromatography mass spectrometry were gamma-terpinene (33.0%), thymol (29.9%), beta-bisabolene (8.9%), p-cymene (8.3%), alpha-terpinene (5.0%), myrcene (4.7%), beta-caryophyllene (4.0%), alpha-thujene (2.7%), camphene (1.3%), carvacrol (1.2%), and alpha-pinene (1.1%). Among these candidates, thymol exhibited complete (100%) repellent activity against female Ae. albopictus, an effect that was confirmed through evaluating the electrophysiological response on the antenna of Ae. albopictus. The effectiveness of a binary 1:2 mixture of thymol and vanillin (0.05:0.1 microl per cm2) was found to be significantly more effective than thymol alone for a period of 120 min. In addition, thymol, alpha-terpinene, and carvacrol showed high larvicidal activity against on the third-stage larvae with LC50 values of 0.9 microl per 100 ml.

  7. On the analysis of parasite effect for Aedes aegypti and Aedes albopictus population

    NASA Astrophysics Data System (ADS)

    Kallista, Meta; Aldila, Dipo; Nuraini, Nuning; Soewono, Edy

    2014-03-01

    It has been reported in some countries that the population of Aedes aegypti has been significantly reduced by the invasion of Aedes albopictus. There has been a hypothesis explaining this phenomenon of which investigated the influence of parasites pathogenesis to the competition between these two mosquito species in the fields. Ascogregarina taiwanensis and Ascogregarina culicis are known as parasites that infect Aedes albopictus and Aedes aegypti, respectively. Several studies have concluded that Ascogregarina taiwanensis caused high fatality for Aedes aegypti larvae, but Ascogregarina culicis was not pathogenic to Aedes albopictus larvae. Therefore, Ascogregarina taiwanensis may contribute to reduce the number of populations Aedes aegypti in the fields. Inspired by these facts, a mathematical model depicting interaction between parasites and mosquitoes is constructed in this paper. In this model are included six dynamic mosquito compartments, i.e. egg, larvae, infected larvae, adult, infected adult and one dynamic compartment for parasite. Derivation of the existence criteria and the stability analysis of parasite-free equilibrium as well as the basic offspring for the model are presented. Numerical simulations for sensitivity analysis indicating the invasive species for variation parameters are shown.

  8. Seasonal Differences in Density But Similar Competitive Impact of Aedes albopictus (Skuse) on Aedes aegypti (L.) in Rio de Janeiro, Brazil

    PubMed Central

    Camara, Daniel Cardoso Portela; Codeço, Claudia Torres; Juliano, Steven A.; Lounibos, L. Philip; Riback, Thais Irene Souza; Pereira, Glaucio Rocha; Honorio, Nildimar Alves

    2016-01-01

    Previous studies have shown that the negative effects of density of Ae. albopictus on Ae. aegypti exceed those of Ae. aegypti on Ae. albopictus for population growth, adult size, survivorship, and developmental rate. This competitive superiority has been invoked to explain the displacement of Ae. aegypti by Ae. albopictus in the southeastern USA. In Brazil, these species coexist in many vegetated suburban and rural areas. We investigated a related, but less-well-studied question: do effects of Ae. albopictus on Ae. aegypti larval development and survival occur under field conditions at realistic densities across multiple seasons in Brazil? We conducted additive competition experiments in a vegetated area of Rio de Janeiro where these species coexist. We tested the hypothesis that Ae. aegypti (the focal species, at a fixed density) suffers negative effects on development and survivorship across a gradient of increasing densities of Ae. albopictus (the associate species) in three seasons. The results showed statistically significant effects of both season and larval density on Ae. aegypti survivorship, and significant effects of season on development rate, with no significant season-density interactions. Densities of Aedes larvae in these habitats differed among seasons by a factor of up to 7x. Overall, Spring was the most favorable season for Ae. aegypti survivorship and development. Results showed that under natural conditions the negative competitive effects of Ae. albopictus on Ae. aegypti were expressed primarily as lower survivorship. Coexistence between Ae. aegypti and Ae. albopictus in vegetated areas is likely affected by seasonal environmental differences, such as detrital resource levels or egg desiccation, which can influence competition between these species. Interactions between these Aedes are important in Brazil, where both species are well established and widely distributed and vector dengue, Zika and chikungunya viruses. PMID:27322537

  9. Drug/Cell-line Browser: interactive canvas visualization of cancer drug/cell-line viability assay datasets.

    PubMed

    Duan, Qiaonan; Wang, Zichen; Fernandez, Nicolas F; Rouillard, Andrew D; Tan, Christopher M; Benes, Cyril H; Ma'ayan, Avi

    2014-11-15

    Recently, several high profile studies collected cell viability data from panels of cancer cell lines treated with many drugs applied at different concentrations. Such drug sensitivity data for cancer cell lines provide suggestive treatments for different types and subtypes of cancer. Visualization of these datasets can reveal patterns that may not be obvious by examining the data without such efforts. Here we introduce Drug/Cell-line Browser (DCB), an online interactive HTML5 data visualization tool for interacting with three of the recently published datasets of cancer cell lines/drug-viability studies. DCB uses clustering and canvas visualization of the drugs and the cell lines, as well as a bar graph that summarizes drug effectiveness for the tissue of origin or the cancer subtypes for single or multiple drugs. DCB can help in understanding drug response patterns and prioritizing drug/cancer cell line interactions by tissue of origin or cancer subtype. DCB is an open source Web-based tool that is freely available at: http://www.maayanlab.net/LINCS/DCB CONTACT: avi.maayan@mssm.edu Supplementary data are available at Bioinformatics online. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  10. Area-Wide Ground Applications of Bacillus thuringiensis var. israelensis for the Control of Aedes albopictus in Residential Neighborhoods: From Optimization to Operation

    PubMed Central

    Williams, Gregory M.; Faraji, Ary; Unlu, Isik; Healy, Sean P.; Farooq, Muhammad; Gaugler, Randy; Hamilton, George; Fonseca, Dina M.

    2014-01-01

    The increasing range of Aedes albopictus, the Asian tiger mosquito, in the USA and the threat of chikungunya and dengue outbreaks vectored by this species have necessitated novel approaches to control this peridomestic mosquito. Conventional methods such as adulticiding provide temporary relief, but fail to manage this pest on a sustained basis. We explored the use of cold aerosol foggers and misting machines for area-wide applications of Bacillus thuringiensis var. israelensis (VectoBac WDG) as a larvicide targeting Aedes albopictus. During 2010–2013 we performed initially open field trials and then 19 operational area-wide applications in urban and suburban residential areas in northeastern USA to test three truck-mounted sprayers at two application rates. Area-wide applications of WDG in open field conditions at 400 and 800 g/ha killed on average 87% of tested larvae. Once techniques were optimized in residential areas, applications with a Buffalo Turbine Mist Sprayer at a rate of 800 g/ha, the best combination, consistently provided over 90% mortality. Importantly, there was no significant decrease in efficacy with distance from the spray line even in blocks of row homes with trees and bushes in the backyards. Under laboratory conditions Bti deposition in bioassay cups during the operational trials resulted in over 6 weeks of residual control. Our results demonstrate that area-wide truck mounted applications of WDG can effectively suppress Ae. albopictus larvae and should be used in integrated mosquito management approaches to control this nuisance pest and disease vector. PMID:25329314

  11. Area-wide ground applications of Bacillus thuringiensis var. israelensis for the control of Aedes albopictus in residential neighborhoods: from optimization to operation.

    PubMed

    Williams, Gregory M; Faraji, Ary; Unlu, Isik; Healy, Sean P; Farooq, Muhammad; Gaugler, Randy; Hamilton, George; Fonseca, Dina M

    2014-01-01

    The increasing range of Aedes albopictus, the Asian tiger mosquito, in the USA and the threat of chikungunya and dengue outbreaks vectored by this species have necessitated novel approaches to control this peridomestic mosquito. Conventional methods such as adulticiding provide temporary relief, but fail to manage this pest on a sustained basis. We explored the use of cold aerosol foggers and misting machines for area-wide applications of Bacillus thuringiensis var. israelensis (VectoBac WDG) as a larvicide targeting Aedes albopictus. During 2010-2013 we performed initially open field trials and then 19 operational area-wide applications in urban and suburban residential areas in northeastern USA to test three truck-mounted sprayers at two application rates. Area-wide applications of WDG in open field conditions at 400 and 800 g/ha killed on average 87% of tested larvae. Once techniques were optimized in residential areas, applications with a Buffalo Turbine Mist Sprayer at a rate of 800 g/ha, the best combination, consistently provided over 90% mortality. Importantly, there was no significant decrease in efficacy with distance from the spray line even in blocks of row homes with trees and bushes in the backyards. Under laboratory conditions Bti deposition in bioassay cups during the operational trials resulted in over 6 weeks of residual control. Our results demonstrate that area-wide truck mounted applications of WDG can effectively suppress Ae. albopictus larvae and should be used in integrated mosquito management approaches to control this nuisance pest and disease vector.

  12. [Detection of Aedes albopictus (Skuse) (Diptera: Culicidae) in the municipality of Istmina, Chocó, Colombia].

    PubMed

    Carvajal, José Joaquín; Honorio, Nildimar Alves; Díaz, Silvia Patricia; Ruiz, Edinso Rafael; Asprilla, Jimmy; Ardila, Susanne; Parra-Henao, Gabriel

    2016-09-01

    Aedes albopictus is widely distributed around the world. Its introduction to the Americas occurred in 1985 and it is considered a potential vector of dengue viruses and one of the principal vectors of chikungunya virus. In Colombia, this species was reported for the first time in Leticia (Amazonas) in 1998, followed by Buenaventura (Valle del Cauca) in 2001, Barrancabermeja (Santander) in 2010, and Medellín (Antioquia) in 2011. So far, this species has been reported in ten departments of the country. Objective: To report the finding of A. albopictus in the city of Istmina, Chocó, and its implications for public health. Materials and methods: In January 2015, we conducted an inspection of immature stages of Aedes spp. in breeding sites in the neighborhoods of San Agustín, Santa Genoveva and Subestación in Istmina, Chocó. The immature stages collected in this municipality were identified at the Unidad de Entomología of the Laboratorio de Salud Pública Departamental de Chocó, and confirmed by the Laboratorio de Entomología, Red Nacional de Laboratorios, Instituto Nacional de Salud, in Bogotá. In January 2015, twelve A. albopictus larvae were found in the breeding sites located in Subestación and San Agustín neighborhoods. The occurrence of A. albopictus in the municipality of Istmina underlines the importance of strengthening continuous entomological surveillance strategies at national and local levels in the country, especially in Istmina and its surrounding municipalities.

  13. History of leukemia-lymphoma cell lines.

    PubMed

    Drexler, Hans G; Macleod, Roderick A F

    2010-08-01

    We outline the near 50-year history of leukemia-lymphoma (LL) cell lines - a key model system in biomedicine. Due to the detailed documentation of their oncogenomic and transcriptional alterations via recent advances in molecular medicine, LL cell lines may be fitted to parent tumors with a degree of precision unattainable in other cancers. We have surveyed the corpus of published LL cell lines and found 637 examples that meet minimum standards of authentication and characterization. Alarmingly, the rate of establishment of new LL cell lines has plummeted over the last decade. Although the main hematopoietic developmental cell types are represented by cell lines, some LL categories stubbornly resist establishment in vitro. The advent of engineering techniques for immortalizing primary human cells that maintain differentiation means the time is ripe for renewed search for in vitro models from un(der)represented hematologic entities. Given their manifold applications in biomedicine, there is little doubt that LL-derived cell lines will continue to play a vital part well into the next half-century as well. © 2010 The Authors. Human Cell © 2010 Japan Human Cell Society.

  14. Projection of climatic suitability for Aedes albopictus Skuse (Culicidae) in Europe under climate change conditions

    NASA Astrophysics Data System (ADS)

    Fischer, Dominik; Thomas, Stephanie Margarete; Niemitz, Franziska; Reineking, Björn; Beierkuhnlein, Carl

    2011-07-01

    During the last decades the disease vector Aedes albopictus ( Ae. albopictus) has rapidly spread around the globe. The spread of this species raises serious public health concerns. Here, we model the present distribution and the future climatic suitability of Europe for this vector in the face of climate change. In order to achieve the most realistic current prediction and future projection, we compare the performance of four different modelling approaches, differentiated by the selection of climate variables (based on expert knowledge vs. statistical criteria) and by the geographical range of presence records (native range vs. global range). First, models of the native and global range were built with MaxEnt and were either based on (1) statistically selected climatic input variables or (2) input variables selected with expert knowledge from the literature. Native models show high model performance (AUC: 0.91-0.94) for the native range, but do not predict the European distribution well (AUC: 0.70-0.72). Models based on the global distribution of the species, however, were able to identify all regions where Ae. albopictus is currently established, including Europe (AUC: 0.89-0.91). In a second step, the modelled bioclimatic envelope of the global range was projected to future climatic conditions in Europe using two emission scenarios implemented in the regional climate model COSMO-CLM for three time periods 2011-2040, 2041-2070, and 2071-2100. For both global-driven models, the results indicate that climatically suitable areas for the establishment of Ae. albopictus will increase in western and central Europe already in 2011-2040 and with a temporal delay in eastern Europe. On the other hand, a decline in climatically suitable areas in southern Europe is pronounced in the Expert knowledge based model. Our projections appear unaffected by non-analogue climate, as this is not detected by Multivariate Environmental Similarity Surface analysis. The generated risk maps

  15. Dispersal of male and female Culex quinquefasciatus and Aedes albopictus mosquitoes using stable isotope enrichment

    PubMed Central

    Roark, E. Brendan; Hamer, Gabriel L.

    2017-01-01

    The dispersal patterns of mosquito vectors are important drivers of vector-borne infectious disease dynamics and understanding movement patterns is pivotal to devise successful intervention strategies. Here, we investigate the dispersal patterns of two globally important mosquito vectors, Aedes albopictus and Culex quinquefasciatus, by marking naturally-occurring larvae with stable isotopes (13C or 15N). Marked individuals were captured with 32 CDC light trap, 32 gravid trap, and 16 BG Sentinel at different locations within two-kilometer radii of six larval habitats enriched with either 13C or 15N. In total, 720 trap nights from July to August 2013 yielded a total of 32,140 Cx. quinquefasciatus and 7,722 Ae. albopictus. Overall, 69 marked female mosquitoes and 24 marked male mosquitoes were captured throughout the study period. The distance that Cx. quinquefasciatus females traveled differed for host-seeking and oviposition-seeking traps, with females seeking oviposition sites traveling further than those seeking hosts. Our analysis suggests that 41% of Cx. quinquefasciatus females that were host-seeking occurred 1–2 kilometer from their respective natal site, while 59% remained within a kilometer of their natal site. In contrast, 59% of Cx. quinquefasciatus females that were seeking oviposition sites occurred between 1–2 kilometer away from their larval habitat, while 15% occurred > 2 kilometer away from their natal site. Our analysis estimated that approximately 100% of Ae. albopictus females remained within 1 km of their respective natal site, with 79% occurring within 250m. In addition, we found that male Ae. albopictus dispersed farther than females, suggesting male-biased dispersal in this Ae. albopictus population. This study provides important insights on the dispersal patterns of two globally relevant vector species, and will be important in planning next generation vector control strategies that mitigate mosquito-borne disease through sterile insect

  16. First report of Aedes (Stegomyia) albopictus (Diptera: Culicidae) in Oran, West of Algeria.

    PubMed

    Benallal, K E; Allal-Ikhlef, A; Benhamouda, K; Schaffner, F; Harrat, Z

    2016-12-01

    The increasing globalisation of trades, human movements and environmental changes facilitate the introduction and the establishment of the invasive Asian tiger mosquito Aedes albopictus outside its native geographical area. Alerted by the complaints about mosquito biting which occurred daytime for the inhabitants of the seaside town Ain Turk (West of Algeria), an entomological survey was conducted in December 2015 to determine the origin of this nuisance. Among the collected mosquitoes, specimens of Ae. albopictus (2 males, 3 females and 3 pupae) were collected. This is the first observation of that invasive mosquito in the west of Algeria which confirms its presence and establishment in Mediterranean Africa. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. First record of Aedes (Stegomyia) albopictus in Belgium.

    PubMed

    Schaffner, Francis; Van Bortel, Wim; Coosemans, Marc

    2004-06-01

    The 1st record of Aedes albopictus in Belgium was made in a village in Oost-Vlaanderen Province. Two preimaginal stages were collected on October 31, 2000, in the used tire stock of a recycling company that imports tires from the USA and Japan. The species has reproduced on site, and local environmental conditions make its establishment possible. Anopheles plumbeus was a common companion species found in tires in high densities.

  18. Aedes albopictus in an area of Misiones, Argentina.

    PubMed

    Schweigmann, Nicolás; Vezzani, Darío; Orellano, Pablo; Kuruc, Jorge; Boffi, Rolando

    2004-02-01

    Until early 1998 the presence of Aedes albopictus had never been detected in Argentina. During April of the same year, few individuals of this species were recorded in 33 breeding sites found in 25 out of 161 inspected houses in the city of Eldorado, Province of Misiones. The homogeneous spatial distribution of the proliferation foci suggests the existence of a generalized infestation in this locality during the study period.

  19. Mapping the spatial distribution of Aedes aegypti and Aedes albopictus.

    PubMed

    Ding, Fangyu; Fu, Jingying; Jiang, Dong; Hao, Mengmeng; Lin, Gang

    2018-02-01

    Mosquito-borne infectious diseases, such as Rift Valley fever, Dengue, Chikungunya and Zika, have caused mass human death with the transnational expansion fueled by economic globalization. Simulating the distribution of the disease vectors is of great importance in formulating public health planning and disease control strategies. In the present study, we simulated the global distribution of Aedes aegypti and Aedes albopictus at a 5×5km spatial resolution with high-dimensional multidisciplinary datasets and machine learning methods Three relatively popular and robust machine learning models, including support vector machine (SVM), gradient boosting machine (GBM) and random forest (RF), were used. During the fine-tuning process based on training datasets of A. aegypti and A. albopictus, RF models achieved the highest performance with an area under the curve (AUC) of 0.973 and 0.974, respectively, followed by GBM (AUC of 0.971 and 0.972, respectively) and SVM (AUC of 0.963 and 0.964, respectively) models. The simulation difference between RF and GBM models was not statistically significant (p>0.05) based on the validation datasets, whereas statistically significant differences (p<0.05) were observed for RF and GBM simulations compared with SVM simulations. From the simulated maps derived from RF models, we observed that the distribution of A. albopictus was wider than that of A. aegypti along a latitudinal gradient. The discriminatory power of each factor in simulating the global distribution of the two species was also analyzed. Our results provided fundamental information for further study on disease transmission simulation and risk assessment. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. EFFECTS OF SUGAR CONCENTRATION ON FECUNDITY, BITING BEHAVIOR AND SURVIVABILITY OF FEMALE AEDES (STEGOMYIA) ALBOPICTUS (SKUSE).

    PubMed

    Aiman, Muhammad; Kassim, Nur Faeza A; Jong, Zheng-Wei; Webb, Cameron E

    2016-11-01

    This study was conducted to better understand the effect of different sucrose concentrations on Aedes albopictus fecundity, biting behavior and survival. Laboratory strain Ae. albopictus females were raised at four different sucrose concentrations (10%, 30%, 50%, and 70%) and their fecundity, host biting on and survival rates were determined. Mosquitoes fed on high (50% or 70%) showed higher mean fecundity rate compared to those on low (10% or 30%) sucrose concentration, and had higher daily biting rate. On the other hand, mosquitoes fed on the low (10% or 30%) sucrose concentrations recorded higher survival rate. These results suggest female mosquitoes deficient in nutrient intake during sugar feeding may regain nutrients needed during blood feeding, whereas those fed on high sucrose concentration have high fecundity due to high biting rate but have low survivability due to low sucrose intake during sugar feeding. Thus, Ae. albopictus females have a capability to regulate their metabolic needs based on sugar nutrient availability.

  1. Efficacy of attractive toxic sugar baits (ATSB) against Aedes albopictus with garlic oil encapsulated in beta-Cyclodextrin as the active ingredient

    USDA-ARS?s Scientific Manuscript database

    We tested the efficacy of attractive toxic sugar bait (ATSB) with garlic oil microencapsulated in beta-cyclodextrin as active ingredient against Aedes albopictus in suburban Haifa, Israel. Two three-acre gardens with high numbers of Ae. albopictus were chosen for perimeter spray treatment with ATSB ...

  2. Interspecific Cross-Mating Between Aedes aegypti and Aedes albopictus Laboratory Strains: Implication of Population Density on Mating Behaviors.

    PubMed

    Marcela, P; Hassan, A Abu; Hamdan, A; Dieng, H; Kumara, T K

    2015-12-01

    Mating behavior between Aedes aegypti and Ae. albopictus, established colony strains were examined under laboratory conditions (30-cm(3) screened cages) for 5 consecutive days. The effect of selected male densities (30, 20, 10) and female density (20) on the number of swarming, mating pairs, eggs produced, and inseminated females were evaluated. Male densities significantly increased swarming behavior, mating pairs, and egg production of heterospecific females, but female insemination was reduced. Aedes aegypti males mate more readily with heterospecific females than do Ae. albopictus males. The current study suggests that Ae. aegypti males were not species-specific in mating, and if released into the field as practiced in genetically modified mosquito techniques, they may mate with both Ae. aegypti and Ae. albopictus females, hence reducing populations of both species by producing infertile eggs.

  3. Cancer stem cell-like cells from a single cell of oral squamous carcinoma cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Felthaus, O.; Department of Oral and Maxillofacial Surgery, University of Regensburg; Ettl, T.

    2011-04-01

    Research highlights: {yields} Four oral squamous cancer cell lines (OSCCL) were analyzed for cancer stem cells (CSCs). {yields} Single cell derived colonies of OSCCL express CSC-marker CD133 differentially. {yields} Monoclonal cell lines showed reduced sensitivity for Paclitaxel. {yields} In situ CD133{sup +} cells are slow cycling (Ki67-) indicating a reduced drug sensitivity. {yields} CD133{sup +} and CSC-like cells can be obtained from single colony forming cells of OSCCL. -- Abstract: Resistance of oral squamous cell carcinomas (OSCC) to conventional chemotherapy or radiation therapy might be due to cancer stem cells (CSCs). The development of novel anticancer drugs requires a simplemore » method for the enrichment of CSCs. CSCs can be enriched from OSCC cell lines, for example, after cultivation in serum-free cell culture medium (SFM). In our study, we analyzed four OSCC cell lines for the presence of CSCs. CSC-like cells could not be enriched with SFM. However, cell lines obtained from holoclone colonies showed CSC-like properties such as a reduced rate of cell proliferation and a reduced sensitivity to Paclitaxel in comparison to cells from the parental lineage. Moreover, these cell lines differentially expressed the CSC-marker CD133, which is also upregulated in OSCC tissues. Interestingly, CD133{sup +} cells in OSCC tissues expressed little to no Ki67, the cell proliferation marker that also indicates reduced drug sensitivity. Our study shows a method for the isolation of CSC-like cell lines from OSCC cell lines. These CSC-like cell lines could be new targets for the development of anticancer drugs under in vitro conditions.« less

  4. Geographic distribution of Aedes aegypti and Aedes albopictus collected from used tires in Vietnam.

    PubMed

    Higa, Yukiko; Yen, Nguyen Thi; Kawada, Hitoshi; Son, Tran Hai; Hoa, Nguyen Thuy; Takagi, Masahiro

    2010-03-01

    The spatial distribution of Aedes aegypti and Aedes albopictus in environmental and geographical zones, e.g., urban-rural, coastal-mountainous, and north-south, was investigated throughout Vietnam. Immature stages were collected from used tires along roads. The effects of regions, seasons, and the degree of urbanization on the density and the frequency were statistically analyzed. Aedes aegypti predominated in the southern and central regions, while Ae. albopictus predominated in the northern region, which may be related to climatic conditions (temperature and rainfall). Larval collection from used tires may be suitable to assess rapidly the current distribution of dengue mosquitoes for estimating health risks and implementing vector control measures.

  5. Vector competence of Italian Aedes albopictus populations for the chikungunya virus (E1-226V).

    PubMed

    Severini, Francesco; Boccolini, Daniela; Fortuna, Claudia; Di Luca, Marco; Toma, Luciano; Amendola, Antonello; Benedetti, Eleonora; Minelli, Giada; Romi, Roberto; Venturi, Giulietta; Rezza, Giovanni; Remoli, Maria Elena

    2018-04-01

    Chikungunya virus (CHIKV) is an emerging arbovirus, belonging to the Togaviridae family, Alphavirus genus, transmitted by Aedes spp. mosquitoes. Since 2007, two different CHIKV strains (E1-226A and E1-226V) have been responsible for outbreaks in European countries, including Italy, sustained by Ae. albopictus mosquitoes. In this study, we assessed the susceptibility to the CHIKV E1-226V, strain responsible for the Italian 2007 outbreak, of eight Ae. albopictus populations collected in Northern, Central, Southern, and Island Italy, by experimental infections. Vector competence was evaluated by estimating infection, dissemination, and transmission rates (IR, DR, TR), through detection of the virus in the bodies, legs plus wings, and saliva, respectively. Additionally, vertical transmission was evaluated by the detection of the virus in the offspring. The results of our study demonstrated that the Italian populations of Ae. albopictus tested were susceptible to CHIKV infection, and can disseminate the virus outside the midgut barrier with high values of IR and DR. Viral infectious RNA was detected in the saliva of three populations from Central, Southern, and Island Italy, also tested for TR and population transmission rate (PTR) values. No progeny of the first and second gonotrophic cycle were positive for CHIKV. This study strongly confirms the role of Ae. albopictus as a potential CHIKV vector in Italy. This may represent a threat, especially considering both the high density of this species, which is widespread throughout the country, and the increasing number of cases of imported arboviruses.

  6. LINE-1 Cultured Cell Retrotransposition Assay

    PubMed Central

    Kopera, Huira C.; Larson, Peter A.; Moldovan, John B.; Richardson, Sandra R.; Liu, Ying; Moran, John V.

    2016-01-01

    Summary The Long INterspersed Element-1 (LINE-1 or L1) retrotransposition assay has facilitated the discovery and characterization of active (i.e., retrotransposition-competent) LINE-1 sequences from mammalian genomes. In this assay, an engineered LINE-1 containing a retrotransposition reporter cassette is transiently transfected into a cultured cell line. Expression of the reporter cassette, which occurs only after a successful round of retrotransposition, allows the detection and quantification of the LINE-1 retrotransposition efficiency. This assay has yielded insight into the mechanism of LINE-1 retrotransposition. It also has provided a greater understanding of how the cell regulates LINE-1 retrotransposition and how LINE-1 retrotransposition impacts the structure of mammalian genomes. Below, we provide a brief introduction to LINE-1 biology and then detail how the LINE-1 retrotransposition assay is performed in cultured mammalian cells. PMID:26895052

  7. Simple Indices Provide Insight to Climate Attributes Delineating the Geographic Range of Aedes albopictus (Diptera: Culicidae) Prior to Worldwide Invasion.

    PubMed

    Mogi, Motoyoshi; Armbruster, Peter; Tuno, Nobuko; Campos, Raúl; Eritja, Roger

    2015-07-01

    Aedes albopictus (Skuse) has expanded its distribution worldwide during the past decades. Despite attempts to explain and predict its geographic occurrence, analyses of the distribution of Ae. albopictus in the context of broad climatic regions (biomes) has not been performed. We analyzed climate conditions at its distribution sites in the range before the worldwide invasions (from the easternmost Hawaii through westernmost Madagascar) by using thermal and aridity-humidity indices descriptive of major biomes. A significant advantage of this approach is that it uses simple indices clearly related to the population dynamics of Ae. albopictus. Although Ae. albopictus has been regarded as a forest species preferring humid climate, in areas with significant human habitation, the distribution sites extended from the perhumid, rain forest zone to the semiarid, steppe zone. This pattern was common from the tropics through the temperate zone. Across the distribution range, there was no seasonal discordance between temperature and precipitation; at sites where winter prevents Ae. albopictus reproduction (monthly means<10°C), precipitation was concentrated in warm months (>10°C) under the Asian summer monsoon. Absence of the species in northern and eastern coastal Australia and eastern coastal Africa was not attributable solely to climate conditions. However, Asia west of the summer monsoon range was climatically unsuitable because of low precipitation throughout the year or in warm months favorable to reproduction (concentration of precipitation in winter). We hypothesized that Ae. albopictus originated in continental Asia under the monsoon climate with distinct dry seasons and hot, wet summer, enabling rapid population growth. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  8. Genome-wide SNPs reveal the drivers of gene flow in an urban population of the Asian Tiger Mosquito, Aedes albopictus.

    PubMed

    Schmidt, Thomas L; Rašić, Gordana; Zhang, Dongjing; Zheng, Xiaoying; Xi, Zhiyong; Hoffmann, Ary A

    2017-10-01

    Aedes albopictus is a highly invasive disease vector with an expanding worldwide distribution. Genetic assays using low to medium resolution markers have found little evidence of spatial genetic structure even at broad geographic scales, suggesting frequent passive movement along human transportation networks. Here we analysed genetic structure of Aedes albopictus collected from 12 sample sites in Guangzhou, China, using thousands of genome-wide single nucleotide polymorphisms (SNPs). We found evidence for passive gene flow, with distance from shipping terminals being the strongest predictor of genetic distance among mosquitoes. As further evidence of passive dispersal, we found multiple pairs of full-siblings distributed between two sample sites 3.7 km apart. After accounting for geographical variability, we also found evidence for isolation by distance, previously undetectable in Ae. albopictus. These findings demonstrate how large SNP datasets and spatially-explicit hypothesis testing can be used to decipher processes at finer geographic scales than formerly possible. Our approach can be used to help predict new invasion pathways of Ae. albopictus and to refine strategies for vector control that involve the transformation or suppression of mosquito populations.

  9. Insecticide susceptibility of Aedes albopictus and Ae. aegypti from Brazil and the Swiss-Italian border region.

    PubMed

    Suter, Tobias; Crespo, Mônica Maria; de Oliveira, Mariana Francelino; de Oliveira, Thaynan Sama Alves; de Melo-Santos, Maria Alice Varjal; de Oliveira, Cláudia Maria Fontes; Ayres, Constância Flávia Junqueira; Barbosa, Rosângela Maria Rodrigues; Araújo, Ana Paula; Regis, Lêda Narcisa; Flacio, Eleonora; Engeler, Lukas; Müller, Pie; Silva-Filha, Maria Helena Neves Lobo

    2017-09-19

    Aedes aegypti and Ae. albopictus are two highly invasive mosquito species, both vectors of several viruses, including dengue, chikungunya and Zika. While Ae. aegypti is the primary vector in the tropics and sub-tropics, Ae. albopictus is increasingly under the public health watch as it has been implicated in arbovirus-transmission in more temperate regions, including continental Europe. Vector control using insecticides is the pillar of most control programmes; hence development of insecticide resistance is of great concern. As part of a Brazilian-Swiss Joint Research Programme we set out to assess whether there are any signs of existing or incipient insecticide resistance primarily against the larvicide Bacillus thuringiensis svar. israelensis (Bti), but also against currently applied and potentially alternative insecticides in our areas, Recife (Brazil) and the Swiss-Italian border region. Following World Health Organization guidelines, dose-response curves for a range of insecticides were established for both colonized and field caught Ae. aegypti and Ae. albopictus. The larvicides included Bti, two of its toxins, Cry11Aa and Cry4Ba, Lysinibacillus sphaericus, Vectomax CG®, a formulated combination of Bti and L. sphaericus, and diflubenzuron. In addition to the larvicides, the Swiss-Italian Ae. albopictus populations were also tested against five adulticides (bendiocarb, dichlorodiphenyltrichloroethane, malathion, permethrin and λ-cyhalothrin). Showing a similar dose-response, all mosquito populations were fully susceptible to the larvicides tested and, in particular, to Bti which is currently used both in Brazil and Switzerland. In addition, there were no signs of incipient resistance against Bti as larvae were equally susceptible to the individual toxins, Cry11Aa and Cry4Ba. The field-caught Swiss-Italian populations were susceptible to the adulticides tested but DDT mortality rates showed signs of reduced susceptibility. The insecticides currently used for

  10. Area-wide management of Aedes albopictus. Part 2: gauging the efficacy of traditional integrated pest control measures against urban container mosquitoes.

    PubMed

    Fonseca, Dina M; Unlu, Isik; Crepeau, Taryn; Farajollahi, Ary; Healy, Sean P; Bartlett-Healy, Kristen; Strickman, Daniel; Gaugler, Randy; Hamilton, George; Kline, Daniel; Clark, Gary G

    2013-12-01

    Aedes (Stegomyia) albopictus (Skuse) is an important disease vector and biting nuisance. During the 2009 active season, six ∼1000-parcel sites were studied, three in urban and three in suburban areas of New Jersey, United States, to examine the efficacy of standard integrated urban mosquito control strategies applied area wide. Active source reduction, larviciding, adulticiding and public education (source reduction through education) were implemented in one site in each county, an education-only approach was developed in a second site and a third site was used as an untreated experimental control. Populations were surveyed weekly with BG-Sentinel traps and ovitraps. A substantial reduction in Ae. albopictus populations was achieved in urban sites, but only modest reductions in suburban sites. Education alone achieved significant reductions in urban adult Ae. albopictus. Egg catches echoed adult catches only in suburban sites. There are significant socioeconomic and climatic differences between urban and suburban sites that impact upon Ae. albopictus populations and the efficacy of the control methods tested. An integrated pest management approach can affect abundances, but labor-intensive, costly source reduction was not enough to maintain Ae. albopictus counts below a nuisance threshold. Nighttime adult population suppression using truck-mounted adulticides can be effective. Area-wide cost-effective strategies are necessary. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.

  11. Generating mammalian stable cell lines by electroporation.

    PubMed

    A Longo, Patti; Kavran, Jennifer M; Kim, Min-Sung; Leahy, Daniel J

    2013-01-01

    Expression of functional, recombinant mammalian proteins often requires expression in mammalian cells (see Single Cell Cloning of a Stable Mammalian Cell Line). If the expressed protein needs to be made frequently, it can be best to generate a stable cell line instead of performing repeated transient transfections into mammalian cells. Here, we describe a method to generate stable cell lines via electroporation followed by selection steps. This protocol will be limited to the CHO dhfr-Urlaub et al. (1983) and LEC1 cell lines, which in our experience perform the best with this method. Copyright © 2013 Elsevier Inc. All rights reserved.

  12. Characterization of a Merkel Cell Polyomavirus-Positive Merkel Cell Carcinoma Cell Line CVG-1.

    PubMed

    Velásquez, Celestino; Amako, Yutaka; Harold, Alexis; Toptan, Tuna; Chang, Yuan; Shuda, Masahiro

    2018-01-01

    Merkel cell polyomavirus (MCV) plays a causal role in ∼80% of Merkel cell carcinomas (MCC). MCV is clonally integrated into the MCC tumor genome, which results in persistent expression of large T (LT) and small T (sT) antigen oncoproteins encoded by the early locus. In MCV-positive MCC tumors, LT is truncated by premature stop codons or deletions that lead to loss of the C-terminal origin binding (OBD) and helicase domains important for replication. The N-terminal Rb binding domain remains intact. MCV-positive cell lines derived from MCC explants have been valuable tools to study the molecular mechanism of MCV-induced Merkel cell carcinogenesis. Although all cell lines have integrated MCV and express truncated LT antigens, the molecular sizes of the LT proteins differ between cell lines. The copy number of integrated viral genome also varies across cell lines, leading to significantly different levels of viral protein expression. Nevertheless, these cell lines share phenotypic similarities in cell morphology, growth characteristics, and neuroendocrine marker expression. Several low-passage MCV-positive MCC cell lines have been established since the identification of MCV. We describe a new MCV-positive MCV cell line, CVG-1, with features distinct from previously reported cell lines. CVG-1 tumor cells grow in more discohesive clusters in loose round cell suspension, and individual cells show dramatic size heterogeneity. It is the first cell line to encode an MCV sT polymorphism resulting in a unique leucine (L) to proline (P) substitution mutation at amino acid 144. CVG-1 possesses a LT truncation pattern near identical to that of MKL-1 cells differing by the last two C-terminal amino acids and also shows an LT protein expression level similar to MKL-1. Viral T antigen knockdown reveals that, like other MCV-positive MCC cell lines, CVG-1 requires T antigen expression for cell proliferation.

  13. Suitability of European climate for the Asian tiger mosquito Aedes albopictus: recent trends and future scenarios

    PubMed Central

    Caminade, Cyril; Medlock, Jolyon M.; Ducheyne, Els; McIntyre, K. Marie; Leach, Steve; Baylis, Matthew; Morse, Andrew P.

    2012-01-01

    The Asian tiger mosquito (Aedes albopictus) is an invasive species that has the potential to transmit infectious diseases such as dengue and chikungunya fever. Using high-resolution observations and regional climate model scenarios for the future, we investigated the suitability of Europe for A. albopictus using both recent climate and future climate conditions. The results show that southern France, northern Italy, the northern coast of Spain, the eastern coast of the Adriatic Sea and western Turkey were climatically suitable areas for the establishment of the mosquito during the 1960–1980s. Over the last two decades, climate conditions have become more suitable for the mosquito over central northwestern Europe (Benelux, western Germany) and the Balkans, while they have become less suitable over southern Spain. Similar trends are likely in the future, with an increased risk simulated over northern Europe and slightly decreased risk over southern Europe. These distribution shifts are related to wetter and warmer conditions favouring the overwintering of A. albopictus in the north, and drier and warmer summers that might limit its southward expansion. PMID:22535696

  14. Nepenthes ampullaria (Nepenthaceae) Pitchers Are Unattractive to Gravid Aedes aegypti and Aedes albopictus (Diptera: Culicidae).

    PubMed

    Chou, Lee Yiung; Dykes, Gary A; Wilson, Robyn F; Clarke, Charles M

    2016-02-01

    Nepenthes pitcher plants are colonized by a variety of specialized arthropods. As Aedes mosquitoes are container breeders, Nepenthes pitchers are a potential candidate oviposition site for vector species, such as Aedes aegypti (L.) and Aedes albopictus (Skuse). However, Aedes spp. are not commonly encountered in Nepenthes pitchers, and the environment inside the pitchers of some species is lethal to them. One exception is Nepenthes ampullaria Jack, whose pitchers are known to be colonized by Ae. albopictus on very rare occasions. Given that Ae. albopictus larvae can survive in N. ampullaria pitcher fluids, we sought to determine why pitcher colonization is rare, testing the hypothesis that gravid Aedes mosquitoes are deterred from ovipositing into container habitats that have similar characteristics to N. ampullaria pitchers. Using plastic ovitraps of different sizes, colors, and with different types of fluids (based on the characteristics of N. ampullaria pitchers), we compared oviposition rates by Aedes mosquitoes in urban and rural areas within the geographical range of N. ampullaria near Kuala Lumpur, Malaysia. Ovitraps that were black and large (>250-ml capacity) accumulated significantly more eggs than ovitraps that were smaller, or green in color. In terms of size and color, small, green ovitraps are analogous to N. ampullaria pitchers, indicating that these pitchers are not particularly attractive to gravid Ae. albopictus. Although Aedes spp. are capable of colonizing N. ampullaria pitchers, the pitchers are relatively unattractive to gravid females and do not represent a significant habitat for larvae of dengue vectors at present. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  15. Enhancement of the BG-sentinel trap with varying number of mice for field sampling of male and female Aedes albopictus mosquitoes.

    PubMed

    Le Goff, Gilbert; Damiens, David; Payet, Laurent; Ruttee, Abdoul-Hamid; Jean, Frédéric; Lebon, Cyrille; Dehecq, Jean-Sébastien; Gouagna, Louis-Clément

    2016-09-22

    Trapping male mosquitoes in the field is essential for the development of area-wide vector control programs with a sterile insect technique (SIT) component. To determine the optimal temporal and spatial release strategy, an estimation of the wild population density and its temporal dynamics is essential. Among the traps available for such data collection, the BG-Sentinel trap developed by the Biogents company uses a combination of visual cues, convection currents and olfactory signals. Although in numerous cases, this trap has shown high efficiency in sampling Aedes albopictus, in some cases low capture rates of Ae. albopictus males were recorded for the BG-sentinel mosquito trap baited with synthetic attractants. The effects of modifying the BG-sentinel trap (by adding one mouse, two or three live mice to the trap) on the efficiency of trapping Ae. albopictus males and females was tested. The experiment was carried out in three distinct areas on La Réunion that have been selected for pilot field testing of the release of sterile male Ae. albopictus mosquitoes. The effect of four types of attractant (including the generic BG-Lure, one mouse or two to three mice) in baited BGS traps was tested with a Latin square design in order to control for the variability of different sampling positions and dates. At the three studied sites, the number of Ae. albopictus adults caught and the proportion of males per trap consistently increased with the number of mice present in the trap. The results from this study suggest that some new attractants derived from, or similar to, mouse odors could be developed and tested in combination with other existing attractive components, such as CO 2 and heat, in order to provide a reliable estimation method for Ae. albopictus adult male abundance in the wild.

  16. Comparison of BG-Sentinel Trap and Oviposition Cups for Aedes aegypti and Aedes albopictus Surveillance in Jacksonville, Florida, USA

    DTIC Science & Technology

    2015-03-01

    albopictus, BG-Sentinel, oviposition cup surveillance, Florida, dengue INTRODUCTION Dengue fever remains a serious mosquitoborne disease in many...state of Florida (FL) in 1986 at a waste tire site in Jacksonville (O’Meara et al. 1992). While the role of Ae. aegypti in the transmission of dengue ...virus is well known, the importance of Ae. albopictus in the transmission of arboviruses in the USA, including dengue , is not clear. Specifically, Cache

  17. Development and evaluation of an attractive self-marking ovitrap to measure dispersal and determine skip oviposition in Aedes albopictus(Diptera:Culicidae) field populations

    USDA-ARS?s Scientific Manuscript database

    Aedes albopictus (Skuse) is a container-breeding mosquito of public health importance. Its oviposition behavior has been assessed in outdoor conditions, but only with laboratory-reared specimens. We used an attractive self-marking oviposition device to assess Ae. albopictus skip oviposition behavi...

  18. The Cellosaurus, a Cell-Line Knowledge Resource

    PubMed Central

    Bairoch, Amos

    2018-01-01

    The Cellosaurus is a knowledge resource on cell lines. It aims to describe all cell lines used in biomedical research. Its scope encompasses both vertebrates and invertebrates. Currently, information for >100,000 cell lines is provided. For each cell line, it provides a wealth of information, cross-references, and literature citations. The Cellosaurus is available on the ExPASy server (https://web.expasy.org/cellosaurus/) and can be downloaded in a variety of formats. Among its many uses, the Cellosaurus is a key resource to help researchers identify potentially contaminated/misidentified cell lines, thus contributing to improving the quality of research in the life sciences. PMID:29805321

  19. Geographic and ecological distribution of the dengue and chikungunya virus vectors Aedes aegypti and Aedes albopictus in three major Cameroonian towns.

    PubMed

    Kamgang, B; Happi, J Y; Boisier, P; Njiokou, F; Hervé, J-P; Simard, F; Paupy, C

    2010-06-01

    Aedes albopictus (Diptera: Culicidae) was first reported in Central Africa in 2000, together with the indigenous mosquito species Aedes aegypti (Diptera: Culicidae). Because Ae. albopictus can also transmit arboviruses, its introduction is a public health concern. We undertook a comparative study in three Cameroonian towns (Sahelian domain: Garoua; equatorial domain: Douala and Yaoundé) in order to document infestation by the two species and their ecological preferences. High and variable levels of pre-imaginal Ae. aegypti and Ae. albopictus infestation were detected. Only Ae. aegypti was encountered in Garoua, whereas both species were found in Douala and Yaoundé, albeit with significant differences in their relative prevalence. Peridomestic water containers were the most strongly colonized and productive larval habitats for both species. No major differences in types of larval habitat were found, but Ae. albopictus preferentially bred in containers containing plant debris or surrounded by vegetation, whereas Ae. aegypti tended to breed in containers located in environments with a high density of buildings. These findings may have important implications for vector control strategies.

  20. Aedes albopictus (Skuse, 1895) (Diptera: Culicidae) in Greece: 13 years of living with the Asian tiger mosquito.

    PubMed

    Badieritakis, Ε; Papachristos, D; Latinopoulos, D; Stefopoulou, Α; Kolimenakis, Α; Bithas, K; Patsoula, Ε; Beleri, S; Maselou, D; Balatsos, G; Michaelakis, Α

    2018-02-01

    Aedes (Stegomyia) albopictus (Skuse, 1895) (Diptera: Culicidae), commonly known as the Asian tiger mosquito, is an invasive mosquito species of public health significance, well established in many countries worldwide. In Greece, it was first recorded in Corfu and Thesprotia between 2003 and 2004. In the following years, further distribution and establishment of Ae. albopictus in Greece have been confirmed in many Regional Units of the country. In the current study, we report the invasion history of Ae. albopictus in Greece, until 2016. The results from the entomological investigation following imported virus cases in 2014, 2015, and 2016 are also included. Moreover, its presence in Greece is demonstrated in a thematic map based on (a) information provided by pest control companies and/or citizens, (b) the official samples sent to Benaki Phytopathological Institute (BPI) and the National School of Public Health (NSPH), and (c) the entomological surveillance conducted by the BPI.

  1. Current and future distribution of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in WHO Eastern Mediterranean Region.

    PubMed

    Ducheyne, Els; Tran Minh, Nhu Nguyen; Haddad, Nabil; Bryssinckx, Ward; Buliva, Evans; Simard, Frédéric; Malik, Mamunur Rahman; Charlier, Johannes; De Waele, Valérie; Mahmoud, Osama; Mukhtar, Muhammad; Bouattour, Ali; Hussain, Abdulhafid; Hendrickx, Guy; Roiz, David

    2018-02-14

    Aedes-borne diseases as dengue, zika, chikungunya and yellow fever are an emerging problem worldwide, being transmitted by Aedes aegypti and Aedes albopictus. Lack of up to date information about the distribution of Aedes species hampers surveillance and control. Global databases have been compiled but these did not capture data in the WHO Eastern Mediterranean Region (EMR), and any models built using these datasets fail to identify highly suitable areas where one or both species may occur. The first objective of this study was therefore to update the existing Ae. aegypti (Linnaeus, 1762) and Ae. albopictus (Skuse, 1895) compendia and the second objective was to generate species distribution models targeted to the EMR. A final objective was to engage the WHO points of contacts within the region to provide feedback and hence validate all model outputs. The Ae. aegypti and Ae. albopictus compendia provided by Kraemer et al. (Sci Data 2:150035, 2015; Dryad Digit Repos, 2015) were used as starting points. These datasets were extended with more recent species and disease data. In the next step, these sets were filtered using the Köppen-Geiger classification and the Mahalanobis distance. The occurrence data were supplemented with pseudo-absence data as input to Random Forests. The resulting suitability and maximum risk of establishment maps were combined into hard-classified maps per country for expert validation. The EMR datasets consisted of 1995 presence locations for Ae. aegypti and 2868 presence locations for Ae. albopictus. The resulting suitability maps indicated that there exist areas with high suitability and/or maximum risk of establishment for these disease vectors in contrast with previous model output. Precipitation and host availability, expressed as population density and night-time lights, were the most important variables for Ae. aegypti. Host availability was the most important predictor in case of Ae. albopictus. Internal validation was assessed

  2. [Health implications of the establishment and spread of Aedes albopictus in Spain].

    PubMed

    Bueno Marí, Rubén; Jiménez Peydró, Ricardo

    2012-01-01

    The spread of Aedes albopictus by Eastern Spain has been constant since its first finding in 2004. Currently the species has been collected in the coastal provinces of Girona, Barcelona, Tarragona, Castellón, Alicante and Murcia. The high synanthropism of the species, together with its anthropophilic behaviour and vectorial capacity to transmit several arboviruses, have led to the return of active transmission cycles of common diseases in the past such as Dengue virus and even the appearance of new tropical viruses as Chikungunya in southern Europe. This manuscript discusses the public health implications of the expected expansion of Ae. albopictus for much of the Iberian Peninsula, in the context of current climatic and sociodemographic situation. Moreover, several issues related with the control of the species in urban and suburban environments are exposed. Finally the preliminary data about other invasive aedines recently collected in the European continent are presented.

  3. Vectorial status of the Asian tiger mosquito Aedes albopictus of La Réunion Island for Zika virus.

    PubMed

    Vazeille, M; Dehecq, J-S; Failloux, A-B

    2018-06-01

    La Réunion Island has been the scene of unusually large epidemics of dengue and chikungunya viruses with Aedes albopictus as the sole vector. After experimental oral infection, Ae. albopictus from La Réunion Island can disseminate both dengue and chikungunya viruses but not the Asian genotype of Zika virus, suggesting a strong midgut barrier to dissemination. Autochthonous transmission of the Asian genotype of Zika virus is improbable on La Réunion Island. © 2017 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of Royal Entomological Society.

  4. Development of a new canine osteosarcoma cell line.

    PubMed

    Séguin, B; Zwerdling, T; McCallan, J L; DeCock, H E V; Dewe, L L; Naydan, D K; Young, A E; Bannasch, D L; Foreman, O; Kent, M S

    2006-12-01

    Establishing a canine osteosarcoma (OSA) cell line can be useful to develop in vivo and in vitro models of OSA. The goal of this study was to develop, characterize and authenticate a new canine OSA cell line and a clone. A cell line and a clone were developed with standard cell culture techniques from a naturally occurring OSA in a dog. The clonal cell line induced a tumour after injection in RAG 1-deficient mouse. Histology was consistent with OSA. The original tumour from the dog and the tumour induced in the mouse were both reactive with vimentin and osteonectin (ON). The parent cell line and clonal cell line were reactive with ON, osteocalcin and alkaline phosphatase. Loss of heterozygosity was found in the same three microsatellite markers in the parent and clonal cell lines, and the tumour tissue grown in the mouse.

  5. Larvicidal Activity of Citrus Limonoids against Aedes albopictus Larvae.

    PubMed

    Bilal, Hazrat; Akram, Waseem; Ali-Hassan, Soaib

    2012-12-01

    Development of insecticide resistance occurred due to the continuous and misuse of synthetic insecticides therefore, the recent study was conducted to explore eco-friendly plant extracts that have some potential to suppress mosquito larval population. WHO recommended mosquito larval bioassay method for insecticide was used while for the analysis of citrus oils for limonin and nomilin content HPLC was used. Among the two citrus cultivars tested as larvicide against Aedes albopictus, valencia late (Citrus sinensis) was the best in terms of LC(50) (297 ppm), % mortality (97%) and LT(50) (18.49 hours) then freutrall early (Citrus reticulate) with LC(50) (377.4 ppm), % mortality (88%) and LT(50) (31 hours), While nomilin gave lowest LC(50) (121.04 ppm) than limonin (382.22 ppm) after 72 hours of exposure. Valencia late also had more limonin and nomilin (377 μg/ml and 21.19 μg/ml) than freutrall early (5.29 μg/ml and 3.89 μg/ml) respectively. Valencia late showed best results in term of LC(50), LT(50) and percentage mortality against Aedes albopictus as it has more amount of nomilin then freutrall early, however further evaluation in the field conditions is required.

  6. Larvicidal Activity of Citrus Limonoids against Aedes albopictus Larvae

    PubMed Central

    Bilal, Hazrat; Akram, Waseem; Ali-Hassan, Soaib

    2012-01-01

    Background: Development of insecticide resistance occurred due to the continuous and misuse of synthetic insecticides therefore, the recent study was conducted to explore eco-friendly plant extracts that have some potential to suppress mosquito larval population. Methods: WHO recommended mosquito larval bioassay method for insecticide was used while for the analysis of citrus oils for limonin and nomilin content HPLC was used. Results: Among the two citrus cultivars tested as larvicide against Aedes albopictus, valencia late (Citrus sinensis) was the best in terms of LC50 (297 ppm), % mortality (97%) and LT50 (18.49 hours) then freutrall early (Citrus reticulate) with LC50 (377.4 ppm), % mortality (88%) and LT50 (31 hours), While nomilin gave lowest LC50 (121.04 ppm) than limonin (382.22 ppm) after 72 hours of exposure. Valencia late also had more limonin and nomilin (377 μg/ml and 21.19 μg/ml) than freutrall early (5.29 μg/ml and 3.89 μg/ml) respectively. Conclusion: Valencia late showed best results in term of LC50, LT50 and percentage mortality against Aedes albopictus as it has more amount of nomilin then freutrall early, however further evaluation in the field conditions is required. PMID:23378967

  7. Assessment of Aedes albopictus (Skuse) clutch size in wild and laboratory populations

    USDA-ARS?s Scientific Manuscript database

    Aedes albopictus (Skuse) is an invasive mosquito species found across the southern U.S. and has undergone range expansion into many northern states. Although primarily pestiferous, it is a capable vector of many disease-causing pathogens. Intra- and interspecific larval competition have been evalu...

  8. Participation of 14-3-3ε and 14-3-3ζ proteins in the phagocytosis, component of cellular immune response, in Aedes mosquito cell lines.

    PubMed

    Trujillo-Ocampo, Abel; Cázares-Raga, Febe Elena; Del Angel, Rosa María; Medina-Ramírez, Fernando; Santos-Argumedo, Leopoldo; Rodríguez, Mario H; Hernández-Hernández, Fidel de la Cruz

    2017-08-01

    Better knowledge of the innate immune system of insects will improve our understanding of mosquitoes as potential vectors of diverse pathogens. The ubiquitously expressed 14-3-3 protein family is evolutionarily conserved from yeast to mammals, and at least two isoforms of 14-3-3, the ε and ζ, have been identified in insects. These proteins have been shown to participate in both humoral and cellular immune responses in Drosophila. As mosquitoes of the genus Aedes are the primary vectors for arboviruses, causing several diseases such as dengue fever, yellow fever, Zika and chikungunya fevers, cell lines derived from these mosquitoes, Aag-2 from Aedes aegypti and C6/36 HT from Aedes albopictus, are currently used to study the insect immune system. Here, we investigated the role of 14-3-3 proteins (ε and ζ isoform) in phagocytosis, the main cellular immune responses executed by the insects, using Aedes spp. cell lines. We evaluated the mRNA and protein expression of 14-3-3ε and 14-3-3ζ in C6/36 HT and Aag-2 cells, and demonstrated that both proteins were localised in the cytoplasm. Further, in C6/36 HT cells treated with a 14-3-3 specific inhibitor we observed a notable modification of cell morphology with filopodia-like structure caused through cytoskeleton reorganisation (co-localization of 14-3-3 proteins with F-actin), more importantly the decrease in Salmonella typhimurium, Staphylococcus aureus and E. coli phagocytosis and reduction in phagolysosome formation. Additionally, silencing of 14-3-3ε and 14-3-3ζ expression by mean of specific DsiRNA confirmed the decreased phagocytosis and phagolysosome formation of pHrodo labelled E. coli and S. aureus bacteria by Aag-2 cells. The 14-3-3ε and 14-3-3ζ proteins modulate cytoskeletal remodelling, and are essential for phagocytosis of Gram-positive and Gram-negative bacteria in Aedes spp. cell lines.

  9. Taming a Tiger in the City: Comparison of Motorized Backpack Applications and Source Reduction Against the Asian Tiger Mosquito, Aedes albopictus

    DTIC Science & Technology

    2014-06-01

    TAMING A TIGER IN THE CITY: COMPARISON OF MOTORIZED BACKPACK APPLICATIONS AND SOURCE REDUCTION AGAINST THE ASIAN TIGER MOSQUITO, AEDES ALBOPICTUS...FONSECA2 AND ARY FARAJOLLAHI2,3,7 ABSTRACT. We evaluated 2 strategies to manage Aedes albopictus: 1) motorized backpack applications and 2) source...the community level. KEY WORDS Bacillus thuringiensis israelensis, Bti, methoprene, hand applications, backpack applications INTRODUCTION Aedes

  10. A field test for competitive effects of Aedes albopictus on A. aegypti in South Florida: differences between sites of coexistence and exclusion?

    PubMed Central

    Juliano, Steven A.; Lounibos, L. Philip; O’Meara, George F.

    2007-01-01

    We tested whether interspecific competition from Aedes albopictus had measurable effects on A. aegypti at the typical numbers of larval mosquitoes found in cemetery vases in south Florida. We also tested whether the effect of interspecific competition from A. albopictus on A. aegypti differed between sites where A. aegypti either persists or went extinct following invasion by A. albopictus. Similar experiments manipulating numbers of A. albopictus in cemetery vases were conducted at three sites of A. aegypti persistence and three sites where A. aegypti was apparently extinct. The experiments were done using numbers of larvae that were determined by observed numbers of larvae for each site, and with resources (leaf detritus) that accumulated in experimental vases placed into each field site. In both the early rainy season (when number of mosquito larvae was low) and the late rainy season (when number of mosquito larvae was high), there was a significant effect of treatment on developmental progress of experimental A. aegypti. In the late rainy season, when numbers of larvae were high, there was also a significant effect of treatment on survivorship of A. aegypti. However, the competition treatment × site type (A. aegypti persists vs extinct) interaction was never significant, indicating that the competitive effect of A. albopictus on A. aegypti did not differ systematically between persistence versus extinction sites. Thus, although competition from A. albopictus is strong under field conditions at all sites, we find no evidence that variation in the impact of interspecific competition is associated with coexistence or exclusion. Interspecific competition among larvae is thus a viable explanation for exclusion or reduction of A. aegypti in south Florida, but variation in the persistence of A. aegypti following invasion does not seem to be primarily a product of variation in the conditions in the aquatic environments of cemetery vases. PMID:15024640

  11. Pyrethroid resistance in Aedes aegypti and Aedes albopictus: Important mosquito vectors of human diseases.

    PubMed

    Smith, Letícia B; Kasai, Shinji; Scott, Jeffrey G

    2016-10-01

    Aedes aegypti and A. albopictus mosquitoes are vectors of important human disease viruses, including dengue, yellow fever, chikungunya and Zika. Pyrethroid insecticides are widely used to control adult Aedes mosquitoes, especially during disease outbreaks. Herein, we review the status of pyrethroid resistance in A. aegypti and A. albopictus, mechanisms of resistance, fitness costs associated with resistance alleles and provide suggestions for future research. The widespread use of pyrethroids has given rise to many populations with varying levels of resistance worldwide, albeit with substantial geographical variation. In adult A. aegypti and A. albopictus, resistance levels are generally lower in Asia, Africa and the USA, and higher in Latin America, although there are exceptions. Susceptible populations still exist in several areas of the world, particularly in Asia and South America. Resistance to pyrethroids in larvae is also geographically widespread. The two major mechanisms of pyrethroid resistance are increased detoxification due to P450-monooxygenases, and mutations in the voltage sensitive sodium channel (Vssc) gene. Several P450s have been putatively associated with insecticide resistance, but the specific P450s involved are not fully elucidated. Pyrethroid resistance can be due to single mutations or combinations of mutations in Vssc. The presence of multiple Vssc mutations can lead to extremely high levels of resistance. Suggestions for future research needs are presented. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. [Establishment of Z-HL16C cell line.].

    PubMed

    Chen, J P; Li, J; Zhao, S L; Tian, J Y; Ye, F

    2006-09-01

    To establish and study the nature and the application of Z-HL16C cell line. The cell line was continuously passed, frozen stored and recovered. Its application was expanded and the cell type was identified. The cell line had an epithelial-cell-like shape, the size appeared uniform, the cell boundary was distinct. It has been continuously passed, frozen stored and recovered for ten years. Its recovery rate was about 90%. It has been proved to be sensitive to the tested viruses which were enteroviruses (Polio, Cox, Echo), influenza viruses, parainfluenzaviruses, adenoviruses, measles virus. This cell line has been identified as a cancerization cell. The cell line Z-HL16C has been stably established, it has a broad spectrum in sensitivity for culturing viruses.

  13. A field survey for Wolbchia and phage WO infections of Aedes albopictus in Guangzhou City, China.

    PubMed

    Zhang, Dongjing; Zhan, Ximei; Wu, Xiansheng; Yang, Xiao; Liang, Gehao; Zheng, Zhantu; Li, Zhuoya; Wu, Yu; Zheng, Xiaoying

    2014-01-01

    Wolbachia are maternal endosymbiotic bacterium, which infect a diverse range of arthropods, ranging from 20 to 76% in nature. They are capable of inducing a wide range of reproductive abnormalities to their hosts, such as cytoplasmic incompatibility (CI), which has been proposed to be used as a tool to modify mosquitoes that are resistant to the development of pathogen, as an alternative vector control strategy. Here, we evaluated the prevalence of Wolbachia and phage WO infections in the field population of Aedes albopictus in Guangzhou City via polymerase chain reaction (PCR) assay using the Wolbachia specific Wolbachia surface protein (wsp) and phage WO orf7 gene primers. Based on the results of PCR and phylogeny analysis, we found that A. albopictus in Guangzhou City were infected with two Wolbachia strains, wAlbA and wAlbB. Phage WO, the virus-infected Wolbachia, was also detected in A. albopictus. One hundred and ten female individuals were screened via PCR, with 109 super-infected with Wolbachia and one sample single-infected with wAlbB strain. And 104 of 113 male individuals were both infected with wAlbA and wAlbB, and nine male samples were found to be infected with wAlbA strain only. The infection rates of phage WO in female and male individuals were 82.73 and 46.02%, respectively. These results showed that the natural Wolbachia and phage WO infections in A. albopictus population in Guangzhou were at a higher frequency at present, indicating that Wolbachia appear to be a better candidate nature resource for biological control insect vectors to reduce vector-borne diseases.

  14. History and updating on the spread of Aedes albopictus in Italy.

    PubMed

    Romi, R

    1995-12-01

    The present distribution in Italy of Aedes albopictus, the Asian mosquito recently introduced, is described. Until now this mosquito species has been established in 8 regions of our country: Liguria, Veneto, Lazio, Lombardia, Emilia-Romagna, Toscana, Piemonte and recently Sardegna. The established populations have been able to overcome the winter season. These populations, certainly imported from temperate areas, have reconfirmed the great adaptation of the species, reaching the high latitude of 46 degrees North of the Padova area in Northern Italy. The origin of the infestation in Veneto region was shown to be strictly related to the importation of used tires and demonstrated at least for one load of tires from USA. The internal trading of tires from Veneto is the source of infestation of the other Italian regions, with the exception of Liguria and Lazio. Since the introduction and the establishment of Ae. albopictus in Italy represent a new problem of public health, a centre for the surveillance and control of this species has been set up at the Department of Parasitology, Istituto Superiore di Sanità, Roma.

  15. Modeling the Environmental Suitability for Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus (Diptera: Culicidae) in the Contiguous United States

    PubMed Central

    Johnson, Tammi L.; Haque, Ubydul; Monaghan, Andrew J.; Eisen, Lars; Hahn, Micah B.; Hayden, Mary H.; Savage, Harry M.; McAllister, Janet; Mutebi, John-Paul; Eisen, Rebecca J.

    2018-01-01

    The mosquitoes Aedes (Stegomyia) aegypti (L.)(Diptera:Culicidae) and Ae. (Stegomyia) albopictus (Skuse) (Diptera:Culicidae) transmit dengue, chikungunya, and Zika viruses and represent a growing public health threat in parts of the United States where they are established. To complement existing mosquito presence records based on discontinuous, non-systematic surveillance efforts, we developed county-scale environmental suitability maps for both species using maximum entropy modeling to fit climatic variables to county presence records from 1960–2016 in the contiguous United States. The predictive models for Ae. aegypti and Ae. albopictus had an overall accuracy of 0.84 and 0.85, respectively. Cumulative growing degree days (GDDs) during the winter months, an indicator of overall warmth, was the most important predictive variable for both species and was positively associated with environmental suitability. The number (percentage) of counties classified as environmentally suitable, based on models with 90 or 99% sensitivity, ranged from 1,443 (46%) to 2,209 (71%) for Ae. aegypti and from 1,726 (55%) to 2,329 (75%) for Ae. albopictus. Increasing model sensitivity results in more counties classified as suitable, at least for summer survival, from which there are no mosquito records. We anticipate that Ae. aegypti and Ae. albopictus will be found more commonly in counties classified as suitable based on the lower 90% sensitivity threshold compared with the higher 99% threshold. Counties predicted suitable with 90% sensitivity should therefore be a top priority for expanded mosquito surveillance efforts while still keeping in mind that Ae. aegypti and Ae. albopictus may be introduced, via accidental transport of eggs or immatures, and potentially proliferate during the warmest part of the year anywhere within the geographic areas delineated by the 99% sensitivity model. PMID:29029153

  16. Niche conservatism of Aedes albopictus and Aedes aegypti - two mosquito species with different invasion histories.

    PubMed

    Cunze, Sarah; Kochmann, Judith; Koch, Lisa K; Klimpel, Sven

    2018-05-16

    Biological invasions have been associated with niche changes; however, their occurrence is still debated. We assess whether climatic niches between native and non-native ranges have changed during the invasion process using two globally spread mosquitoes as model species, Aedes albopictus and Aedes aegypti. Considering the different time spans since their invasions (>300 vs. 30-40 years), niche changes were expected to be more likely for Ae. aegypti than for Ae. albopictus. We used temperature and precipitation variables as descriptors for the realized climatic niches and different niche metrics to detect niche dynamics in the native and non-native ranges. High niche stability, therefore, no niche expansion but niche conservatism was revealed for both species. High niche unfilling for Ae. albopictus indicates a great potential for further expansion. Highest niche occupancies in non-native ranges occurred either under more temperate (North America, Europe) or tropical conditions (South America, Africa). Aedes aegypti has been able to fill its native climatic niche in the non-native ranges, with very low unfilling. Our results challenge the assumption of rapid evolutionary change of climatic niches as a requirement for global invasions but support the use of native range-based niche models to project future invasion risk on a large scale.

  17. Co-breeding Association of Aedes albopictus (Skuse) and Aedes aegypti (Linnaeus) (Diptera: Culicidae) in Relation to Location and Container Size

    PubMed Central

    Hashim, Nur Aida; Ahmad, Abu Hassan; Talib, Anita; Athaillah, Farida; Krishnan, Kumara Thevan

    2018-01-01

    The occurrence of major outbreaks of dengue, and other vector borne diseases such as chikungunya and zika in tropical and subtropical regions has rendered control of the diseases a top-priority for many affected countries including Malaysia. Control of the mosquito vectors Aedes aegypti and Aedes albopictus through the reduction of breeding sites and the application of insecticides to kill immature forms and adults are the main control efforts to combat these diseases. The present study describes the association between Ae. albopictus and Ae. aegypti in shared breeding sites. This study is important given that any measure taken against one species may affect the other. A yearlong larval survey was conducted in four dengue endemic areas of Penang Island. Sorenson’s coefficient index indicated that no association between number of the immatures of the two species regardless of container size and study location. Therefore, the mean number Ae. albopictus immature was not decreased in the presence of Ae. aegypti in shared breeding container. However Ae. aegypti appeared to prefer breeding in habitats not occupied by Ae. albopictus, the two species sharing breeding sites only where available containers were limited. In control efforts, eliminating the preferred breeding containers for one species might not affect or reduce the population of the other species. PMID:29644025

  18. Comparison of BG-Sentinel® Trap and Oviposition Cups for Aedes aegypti and Aedes albopictus Surveillance in Jacksonville, Florida, USA.

    PubMed

    Wright, Jennifer A; Larson, Ryan T; Richardson, Alec G; Cote, Noel M; Stoops, Craig A; Clark, Marah; Obenauer, Peter J

    2015-03-01

    The BG-Sentinel® (BGS) trap and oviposition cups (OCs) have both proven effective in the surveillance of Aedes species. This study aimed to determine which of the 2 traps could best characterize the relative population sizes of Aedes albopictus and Aedes aegypti in an urban section of Jacksonville, FL. Until 1986, Ae. aegypti was considered the dominant container-breeding species in urban northeastern Florida. Since the introduction of Ae. albopictus, Ae. aegypti has become almost completely extirpated. In 2011, a resurgence of Ae. aegypti was detected in the urban areas of Jacksonville; thus this study initially set out to determine the extent of Ae. aegypti reintroduction to the area. We determined that the BGS captured a greater number of adult Ae. aegypti than Ae. albopictus, while OCs did not monitor significantly different numbers of either species, even in areas where the BGS traps suggested a predominance of one species over the other. Both traps were effective at detecting Aedes spp.; however, the BGS proved more diverse by detecting over 20 other species as well. Our results show that in order to accurately determine vectorborne disease threats and the impact of control operations on these 2 species, multiple trapping techniques should be utilized when studying Ae. aegypti and Ae. albopictus population dynamics.

  19. Seasonal Synchronization of Diapause Phases in Aedes albopictus (Diptera: Culicidae).

    PubMed

    Lacour, Guillaume; Chanaud, Lionel; L'Ambert, Grégory; Hance, Thierry

    2015-01-01

    In temperate areas, population dynamics of the invasive Asian tiger mosquito Aedes albopictus are strongly affected by winter. The work we present here analyzes the adaptive synchronization of the diapause process in the wintry generation of A. albopictus, where the egg stage is exposed to adverse winter conditions. The seasonal pattern of egg laying activity of a French Mediterranean population of the Asian tiger mosquito was monitored weekly for 2 years with ovitraps. The field diapause incidence and the critical photoperiod (CPP, i.e. the maternal day length inducing diapause in 50% of the eggs), were determined by hatching experiments on the collected eggs. The period of diapause termination was estimated by a field survey of the first hatchings for both years. The CPP is equal to 13.5 hours of light and occurs in the field on the 25th of August. Thus, it is on September 11th, 17 days after the CPP, that 50% of the eggs are in a prediapause stage in the field. The egg diapause rate increases rapidly during September, whereas the mean number of eggs laid decreases sharply after mid-September. Surprisingly, after having reached a peak of 95% at the end of September, from mid-October the diapause incidence declined and stayed below 50%. Indeed, both years the diapause initiates before the rapid decrease of the environmental temperature. This leaves a sufficient period of time to the complete development of one generation of A. albopictus with effective induction of diapause in the laid eggs. The very first larvae hatched were sampled both years in the first half of March. With 20 to 26 weeks in the egg stage and about 7 weeks in the larval stages, the first annual generation spends a long time in immature stages. On a practical point of view, this long development time represents a wide window for eggs and larvae control in early spring.

  20. The transcriptional diversity of 25 Drosophila cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cherbas, Lucy; Willingham, Aarron; Zhang, Dayu

    2010-12-22

    Drosophila melanogaster cell lines are important resources for cell biologists. In this article, we catalog the expression of exons, genes, and unannotated transcriptional signals for 25 lines. Unannotated transcription is substantial (typically 19% of euchromatic signal). Conservatively, we identify 1405 novel transcribed regions; 684 of these appear to be new exons of neighboring, often distant, genes. Sixty-four percent of genes are expressed detectably in at least one line, but only 21% are detected in all lines. Each cell line expresses, on average, 5885 genes, including a common set of 3109. Expression levels vary over several orders of magnitude. Major signalingmore » pathways are well represented: most differentiation pathways are ‘‘off’’ and survival/growth pathways ‘‘on.’’ Roughly 50% of the genes expressed by each line are not part of the common set, and these show considerable individuality. Thirty-one percent are expressed at a higher level in at least one cell line than in any single developmental stage, suggesting that each line is enriched for genes characteristic of small sets of cells. Most remarkable is that imaginal disc-derived lines can generally be assigned, on the basis of expression, to small territories within developing discs. These mappings reveal unexpected stability of even fine-grained spatial determination. No two cell lines show identical transcription factor expression. We conclude that each line has retained features of an individual founder cell superimposed on a common ‘‘cell line‘‘ gene expression pattern. We report the transcriptional profiles of 25 Drosophila melanogaster cell lines, principally by whole-genome tiling microarray analysis of total RNA, carried out as part of the modENCODE project. The data produced in this study add to our knowledge of the cell lines and of the Drosophila transcriptome in several ways. We summarize the expression of previously annotated genes in each of the 25 lines

  1. Comparison of steroid receptors from the androgen responsive DDT1 cell line and the nonresponsive HVP cell line.

    PubMed

    Norris, J S; Kohler, P O

    1978-01-01

    Two hamster cell lines have been isolated from androgen target tissue. The DDT1 cells derived from ductus deferens tissue exhibit a growth response to androgens, while the HVP cells derived from ventral prostate are androgen unresponsive. Both cell lines contain androgen receptors, that are similar when compared by kinetic methods, sedimentation velocity, chromatographic procedures or nuclear translocation ability. The forms of the high salt extracted nuclear receptors are indistinguishable chromatographically. Therefore, we postulate that the lesion preventing androgen induced growth in the HVP cell line is subseqent to nuclear translocation of the steroid receptor complex.

  2. Two Chikungunya isolates from the outbreak of La Reunion (Indian Ocean) exhibit different patterns of infection in the mosquito, Aedes albopictus.

    PubMed

    Vazeille, Marie; Moutailler, Sara; Coudrier, Daniel; Rousseaux, Claudine; Khun, Huot; Huerre, Michel; Thiria, Julien; Dehecq, Jean-Sébastien; Fontenille, Didier; Schuffenecker, Isabelle; Despres, Philippe; Failloux, Anna-Bella

    2007-11-14

    A Chikungunya (CHIK) outbreak hit La Réunion Island in 2005-2006. The implicated vector was Aedes albopictus. Here, we present the first study on the susceptibility of Ae. albopictus populations to sympatric CHIKV isolates from La Réunion Island and compare it to other virus/vector combinations. We orally infected 8 Ae. albopictus collections from La Réunion and 3 from Mayotte collected in March 2006 with two Chikungunya virus (CHIKV) from La Réunion: (i) strain 05.115 collected in June 2005 with an Alanine at the position 226 of the glycoprotein E1 and (ii) strain 06.21 collected in November 2005 with a substitution A226V. Two other CHIKV isolates and four additional mosquito strains/species were also tested. The viral titer of the infectious blood-meal was 10(7) plaque forming units (pfu)/mL. Dissemination rates were assessed by immunofluorescent staining on head squashes of surviving females 14 days after infection. Rates were at least two times higher with CHIKV 06.21 compared to CHIKV 05.115. In addition, 10 individuals were analyzed every day by quantitative RT-PCR. Viral RNA was quantified on (i) whole females and (ii) midguts and salivary glands of infected females. When comparing profiles, CHIKV 06.21 produced nearly 2 log more viral RNA copies than CHIKV 05.115. Furthermore, females infected with CHIKV 05.115 could be divided in two categories: weakly susceptible or strongly susceptible, comparable to those infected by CHIKV 06.21. Histological analysis detected the presence of CHIKV in salivary glands two days after infection. In addition, Ae. albopictus from La Réunion was as efficient vector as Ae. aegypti and Ae. albopictus from Vietnam when infected with the CHIKV 06.21. Our findings support the hypothesis that the CHIK outbreak in La Réunion Island was due to a highly competent vector Ae. albopictus which allowed an efficient replication and dissemination of CHIKV 06.21.

  3. Two Chikungunya Isolates from the Outbreak of La Reunion (Indian Ocean) Exhibit Different Patterns of Infection in the Mosquito, Aedes albopictus

    PubMed Central

    Vazeille, Marie; Moutailler, Sara; Coudrier, Daniel; Rousseaux, Claudine; Khun, Huot; Huerre, Michel; Thiria, Julien; Dehecq, Jean-Sébastien; Fontenille, Didier; Schuffenecker, Isabelle; Despres, Philippe; Failloux, Anna-Bella

    2007-01-01

    Background A Chikungunya (CHIK) outbreak hit La Réunion Island in 2005–2006. The implicated vector was Aedes albopictus. Here, we present the first study on the susceptibility of Ae. albopictus populations to sympatric CHIKV isolates from La Réunion Island and compare it to other virus/vector combinations. Methodology and Findings We orally infected 8 Ae. albopictus collections from La Réunion and 3 from Mayotte collected in March 2006 with two Chikungunya virus (CHIKV) from La Réunion: (i) strain 05.115 collected in June 2005 with an Alanine at the position 226 of the glycoprotein E1 and (ii) strain 06.21 collected in November 2005 with a substitution A226V. Two other CHIKV isolates and four additional mosquito strains/species were also tested. The viral titer of the infectious blood-meal was 107 plaque forming units (pfu)/mL. Dissemination rates were assessed by immunofluorescent staining on head squashes of surviving females 14 days after infection. Rates were at least two times higher with CHIKV 06.21 compared to CHIKV 05.115. In addition, 10 individuals were analyzed every day by quantitative RT-PCR. Viral RNA was quantified on (i) whole females and (ii) midguts and salivary glands of infected females. When comparing profiles, CHIKV 06.21 produced nearly 2 log more viral RNA copies than CHIKV 05.115. Furthermore, females infected with CHIKV 05.115 could be divided in two categories: weakly susceptible or strongly susceptible, comparable to those infected by CHIKV 06.21. Histological analysis detected the presence of CHIKV in salivary glands two days after infection. In addition, Ae. albopictus from La Réunion was as efficient vector as Ae. aegypti and Ae. albopictus from Vietnam when infected with the CHIKV 06.21. Conclusions Our findings support the hypothesis that the CHIK outbreak in La Réunion Island was due to a highly competent vector Ae. albopictus which allowed an efficient replication and dissemination of CHIKV 06.21. PMID:18000540

  4. Global risk mapping for major diseases transmitted by Aedes aegypti and Aedes albopictus

    PubMed Central

    Leta, Samson; Beyene, Tariku Jibat; De Clercq, Eva M.; Amenu, Kebede; Revie, Crawford W.; Kraemer, Moritz U G

    2018-01-01

    Objectives The objective of this study was to map the global risk of the major arboviral diseases transmitted by Aedes aegypti and Aedes albopictus by identifying areas where the diseases are reported, either through active transmission or travel-related outbreaks, as well as areas where the diseases are not currently reported but are nonetheless suitable for the vector. Methods Data relating to five arboviral diseases (Zika, dengue fever, chikungunya, yellow fever, and Rift Valley fever (RVF)) were extracted from some of the largest contemporary databases and paired with data on the known distribution of their vectors, A. aegypti and A. albopictus. The disease occurrence data for the selected diseases were compiled from literature dating as far back as 1952 to as recent as 2017. The resulting datasets were aggregated at the country level, except in the case of the USA, where state-level data were used. Spatial analysis was used to process the data and to develop risk maps. Results Out of the 250 countries/territories considered, 215 (86%) are potentially suitable for the survival and establishment of A. aegypti and/or A. albopictus. A. albopictus has suitability foci in 197 countries/territories, while there are 188 that are suitable for A. aegypti. There is considerable variation in the suitability range among countries/territories, but many of the tropical regions of the world provide high suitability over extensive areas. Globally, 146 (58.4%) countries/territories reported at least one arboviral disease, while 123 (49.2%) reported more than one of the above diseases. The overall numbers of countries/territories reporting autochthonous vector-borne occurrences of Zika, dengue, chikungunya, yellow fever, and RVF, were 85, 111, 106, 43, and 39, respectively. Conclusions With 215 countries/territories potentially suitable for the most important arboviral disease vectors and more than half of these reporting cases, arboviral diseases are indeed a global public

  5. Global risk mapping for major diseases transmitted by Aedes aegypti and Aedes albopictus.

    PubMed

    Leta, Samson; Beyene, Tariku Jibat; De Clercq, Eva M; Amenu, Kebede; Kraemer, Moritz U G; Revie, Crawford W

    2018-02-01

    The objective of this study was to map the global risk of the major arboviral diseases transmitted by Aedes aegypti and Aedes albopictus by identifying areas where the diseases are reported, either through active transmission or travel-related outbreaks, as well as areas where the diseases are not currently reported but are nonetheless suitable for the vector. Data relating to five arboviral diseases (Zika, dengue fever, chikungunya, yellow fever, and Rift Valley fever (RVF)) were extracted from some of the largest contemporary databases and paired with data on the known distribution of their vectors, A. aegypti and A. albopictus. The disease occurrence data for the selected diseases were compiled from literature dating as far back as 1952 to as recent as 2017. The resulting datasets were aggregated at the country level, except in the case of the USA, where state-level data were used. Spatial analysis was used to process the data and to develop risk maps. Out of the 250 countries/territories considered, 215 (86%) are potentially suitable for the survival and establishment of A. aegypti and/or A. albopictus. A. albopictus has suitability foci in 197 countries/territories, while there are 188 that are suitable for A. aegypti. There is considerable variation in the suitability range among countries/territories, but many of the tropical regions of the world provide high suitability over extensive areas. Globally, 146 (58.4%) countries/territories reported at least one arboviral disease, while 123 (49.2%) reported more than one of the above diseases. The overall numbers of countries/territories reporting autochthonous vector-borne occurrences of Zika, dengue, chikungunya, yellow fever, and RVF, were 85, 111, 106, 43, and 39, respectively. With 215 countries/territories potentially suitable for the most important arboviral disease vectors and more than half of these reporting cases, arboviral diseases are indeed a global public health threat. The increasing proportion of

  6. Characterization of stem-like cells in a new astroblastoma cell line

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Coban, Esra Aydemir; Kasikci, Ezgi; Karatas, Omer Faruk

    Cell lines established from tumors are the most commonly used models in cancer research, and their use in recent years has enabled a greater understanding of the biology of cancer and the means to develop effective treatment strategies. Astroblastomas are uncommon neuroepithelial tumors of glial origin, predominantly affecting young people, mainly teenagers and children, predominantly females. To date, only a single study has reported that astroblastomas contain a large number of neural stem-like cells, which had only a partial proliferation capacity and differentiation. Our objective was to establish an astroblastoma cell line to investigate the presence of astroblastic cells andmore » cancer stem-like cells. The migratory and invasion abilities of the cells were quantified with invasion and migration assays and compared to a glioblastoma cell line. The presence of stem cells was detected with surface-marker analysis by using flow cytometry, and measuring the differentiation ability with a differentiation assay and the self-renewal capacity with a sphere-forming assay. These characteristics may determine whether this novel cell line is a model for astroblastomas that may have stem-cell characteristics. With this novel cell line, scientists can investigate the molecular pathways underlying astroblastomas and develop new therapeutic strategies for patients with these tumors. - Highlights: • An establishment of a novel astroblastoma cell line was proposed. • The presence of astroblastic cells and cancer stem-like cells was investigated. • The molecular pathways underlying astroblastomas may be investigated. • New therapeutic strategies for patients with astroblastoma may be developed.« less

  7. Ontological representation, integration, and analysis of LINCS cell line cells and their cellular responses.

    PubMed

    Ong, Edison; Xie, Jiangan; Ni, Zhaohui; Liu, Qingping; Sarntivijai, Sirarat; Lin, Yu; Cooper, Daniel; Terryn, Raymond; Stathias, Vasileios; Chung, Caty; Schürer, Stephan; He, Yongqun

    2017-12-21

    Aiming to understand cellular responses to different perturbations, the NIH Common Fund Library of Integrated Network-based Cellular Signatures (LINCS) program involves many institutes and laboratories working on over a thousand cell lines. The community-based Cell Line Ontology (CLO) is selected as the default ontology for LINCS cell line representation and integration. CLO has consistently represented all 1097 LINCS cell lines and included information extracted from the LINCS Data Portal and ChEMBL. Using MCF 10A cell line cells as an example, we demonstrated how to ontologically model LINCS cellular signatures such as their non-tumorigenic epithelial cell type, three-dimensional growth, latrunculin-A-induced actin depolymerization and apoptosis, and cell line transfection. A CLO subset view of LINCS cell lines, named LINCS-CLOview, was generated to support systematic LINCS cell line analysis and queries. In summary, LINCS cell lines are currently associated with 43 cell types, 131 tissues and organs, and 121 cancer types. The LINCS-CLO view information can be queried using SPARQL scripts. CLO was used to support ontological representation, integration, and analysis of over a thousand LINCS cell line cells and their cellular responses.

  8. Failure of Aedes albopictus to overwinter following introduction and seasonal establishment at a tire recycling plant in the northeastern USA.

    PubMed

    Andreadis, Theodore G

    2009-03-01

    In July 2006, an introduction of the Asian tiger mosquito, Aedes albopictus, was documented for the first time at a commercial tire recycling plant in northeastern Connecticut, USA. The introduction likely occurred via transport of infested tires originating from northern New Jersey or metropolitan New York City. Efforts were made to determine seasonal establishment and overwintering success by assessing adult biting and oviposition activity in the surrounding woodlands. The first adult female was collected in a CO2-baited Mosquito Magnet Liberty trap within the confines of the tire plant during the week of July 28. Additional females were collected intermittently thereafter through October 16. Host-seeking female Ae. albopictus attempting to alight on human subjects and larvae hatching from eggs collected in ovitraps placed in the woodlands surrounding the tire plant were detected weekly from August 21 through October 2, denoting seasonal establishment in the adjoining woodlands. However, no larvae of Ae. albopictus were recovered from eggs collected in ovitraps that were placed in the surrounding woodlands or in traps placed 1.0-1.6 km away, nor were any host-seeking females detected by human subjects the following season (July to October 2007), indicating that the species did not survive winter conditions to enable successful colonization. The failure of Ae. albopictus to overwinter and establish itself in the forested woodlands following several weeks of seasonal breeding and oviposition during the summer and early fall were most likely due to winter egg mortality, interspecific competition from Aedes triseriatus and Aedes japonicus, and/or other ecological barriers. Permanent establishment of Ae. albopictus in New England is unlikely despite the recurring importation of infested used tires into recycling facilities. However, continued monitoring of such facilities for potential reinvasion is warranted especially in urban/suburban environs where global warming

  9. Coexistence of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Peninsular Florida Two Decades After Competitive Displacements.

    PubMed

    Lounibos, L Philip; Bargielowski, Irka; Carrasquilla, María Cristina; Nishimura, Naoya

    2016-11-01

    The spread of Aedes albopictus (Skuse) eastward in the mid-1980s from its initial establishment in Houston, TX, was associated with rapid declines and local disappearances of Aedes aegypti (L.) in Gulf Coast states and Florida where annual larval surveillance during the early 1990s described temporal and spatial patterns of competitive displacements in cemeteries and tire shops. Approximately 20 yr later in 2013-2014, we re-visited former collection sites and sampled aquatic immatures of these two species from tire shops in 10 cities on State Route 441 and from 9 cemeteries from Lakeland to Miami in southwest Florida. In the recent samples Ae. aegypti was recovered from three central Florida cities where it had not been detected in 1994, but its northern limit on Rte. 441, Apopka, did not change. Other evidence, such as trends at a few cemeteries, suggested a moderate resurgence of this species since 1994. Cage experiments that exposed female progeny of Ae. aegypti from recent Florida collection sites to interspecific mating by Ae. albopictus males showed that females from coexistence sites had evolved resistance to cross-mating, but Ae. aegypti from sites with no Ae. albopictus were relatively susceptible to satyrization. Habitat classifications of collection sites were reduced by principal component (PC) analysis to four variables that accounted for > 99% of variances; PCs with strong positive loadings for tree cover and ground vegetation were associated with collection sites yielding only Ae. albopictus Within the coexistence range of the two species, the numbers of Ae. aegypti among total Aedes collected were strongly correlated in stepwise logistic regression models with two habitat-derived PCs, distance from the coast, and annual rainfall and mean maximum temperatures at the nearest weather station. Subtle increases in the range of Ae. aegypti since its previous displacements are interpreted in the context of the evolution of resistance to mating

  10. The Genetic Diversity, Haplotype Analysis, and Phylogenetic Relationship of Aedes albopictus (Diptera: Culicidae) Based on the Cytochrome Oxidase 1 Marker: A Malaysian Scenario.

    PubMed

    Ismail, Nurul-Ain; Adilah-Amrannudin, Nurul; Hamsidi, Mayamin; Ismail, Rodziah; Dom, Nazri Che; Ahmad, Abu Hassan; Mastuki, Mohd Fahmi; Camalxaman, Siti Nazrina

    2017-11-07

    The global expansion of Ae. albopictus from its native range in Southeast Asia has been implicated in the recent emergence of dengue endemicity in Malaysia. Genetic variability studies of Ae. albopictus are currently lacking in the Malaysian setting, yet are crucial to enhancing the existing vector control strategies. The study was conducted to establish the genetic variability of maternally inherited mitochondrial DNA encoding for cytochrome oxidase subunit 1 (CO1) gene in Ae. albopictus. Twelve localities were selected in the Subang Jaya district based on temporal indices utilizing 120 mosquito samples. Genetic polymorphism and phylogenetic analysis were conducted to unveil the genetic variability and geographic origins of Ae. albopictus. The haplotype network was mapped to determine the genealogical relationship of sequences among groups of population in the Asian region. Comparison of Malaysian CO1 sequences with sequences derived from five Asian countries revealed genetically distinct Ae. albopictus populations. Phylogenetic analysis revealed that all sequences from other Asian countries descended from the same genetic lineage as the Malaysian sequences. Noteworthy, our study highlights the discovery of 20 novel haplotypes within the Malaysian population which to date had not been reported. These findings could help determine the genetic variation of this invasive species, which in turn could possibly improve the current dengue vector surveillance strategies, locally and regionally. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  11. Surface receptors on human haematopoietic cell lines.

    PubMed Central

    Huber, C; Sundström, C; Nilsson, K; Wigzell, H

    1976-01-01

    The expression of complement receptors, of Fc receptors, of SRBC receptors and of S-Ig was investigated on human haematopoietic cell lines of proved malignant derivation. According to their origin and to a panel of phenotypic markers these lines have been classified into lymphoma lines, myeloma lines and leukemia lines. Results were compared with those obtained on non-malignant EBV carrying lymphoblastoid cell lines (LCL). Among the lymphoid cell lines the LCL showed a pattern of B-lymphocyte surface markers, i.e. surface immunoglobulins, C3 receptors but low density of Fc receptors. The non-Burkitt lymphoma lines bore in varying degree these B-lymphocyte markers. The lines U-698 M and DG-75 were exceptional in having only surface immunoglobulin. The Burkitt lymphoma lines had all B-lymphocyte markers. The myeloma lines differed from the lymphoid lines in lacking C3 and Fc receptors and showed only trace amounts of surface immunoglobulins. In contrast to lymphoid and myeloma lines, the leukaemia lines were completely lacking surface immunoglobulins, but showed C3 and Fc receptors in variable densities. On line, the ALL derived line MOLT-3 showed the capacity to spontaneous rosette formation with SRBC. The findings that LCL presented a homogeneous pattern of B-lymphocyte surface markers may be of value in order to discriminate between these lines and lines derived from haematopoietic malignancies other than Burkitt lymphomas. PMID:963908

  12. The effect of Piper aduncum Linn. (Family: Piperaceae) essential oil as aerosol spray against Aedes aegypti (L.) and Aedes albopictus Skuse.

    PubMed

    Misni, Norashiqin; Othman, Hidayatulfathi; Sulaiman, Sallehudin

    2011-08-01

    The bioefficacy of Piper aduncum L. essential oil formulated in aerosol cans was evaluated against Aedes aegypti and Aedes albopictus in a simulated room. The aerosol spray test was based on the Malaysian test standard for aerosol (MS 1221:1991UDC 632.982.2 modified from WHO 2009 methodology) and examined the knockdown effect within 20 minutes of exposure. Mortality rate after 24 hour of holding period was also determined. A commercial aerosol spray (0.09% prallethrin 0.05% d-phenothrin) was also tested as a comparison. Our results showed that the knockdown effect of the commercial aerosol spray and P. aduncum essential oil spray (8% and 10% concentrations) was significantly higher in Ae. albopictus adult females, when compared with that of Ae. aegypti adult females (P<0.05). There was a significant difference in knockdown between commercial aerosol spray and essential oil spray for both Aedes spp. (P<0.05). The essential oil induced significantly higher mortality in Ae. aegypti (80%) than in Ae. albopictus (71.6%) (P<0.05). The commercial aerosol spray caused 97.7% and 86.5% mortality against Ae. aegypti and Ae. albopictus respectively (P<0.05). Based on these data, P. aduncum essential oil has the potential to be used as an aerosol spray against Aedes spp.

  13. Chemical composition and assessment of larvicidal and repellent capacity of 14 Lamiaceae essential oils against Aedes albopictus.

    PubMed

    Giatropoulos, Athanassios; Kimbaris, Athanasios; Michaelakis, Αntonios; Papachristos, Dimitrios P; Polissiou, Moschos G; Emmanouel, Nickolaos

    2018-06-01

    In the current laboratory study, 14 essential oils (EOs) derived from 12 Lamiaceae plant species and their major components were screened for their larvicidal and repellent properties against Aedes albopictus, an invasive mosquito species of great medical importance. The results of toxicity bioassays revealed that the EOs from Thymus vulgaris, Ocimum basilicum, Origanum dictamnus, Origanum majorana, and Origanum vulgare, as well as their major components (terpenes), namely thymol, carvacrol, p-cymene, and γ-terpinene exerted the highest larvicidal effect. Essential oils from Mellisa officinalis, Origanum dictamus, Mentha spicata (chem. piperitenone epoxide), Origanum majorana, and Satureja thymbra were the most potent repellents, with the last two assigned as the best ones. Among the terpenes tested, piperitenone epoxide, carvacrol, thymol, and piperitenone provided the highest level of protection against Ae. albopictus adults. Chemical analysis revealed the presence of a high number of terpenes in the EOs, while in most cases, the biological action of the tested EOs and their major components was in consistency. The most effective EOs and terpenes that were identified through the current laboratory bioassays could be used as alternative agents to control larvae and repel adults of Ae. albopictus.

  14. La Crosse Encephalitis Virus Infection in Field-Collected Aedes albopictus, Aedes japonicus, and Aedes triseriatus in Tennessee.

    PubMed

    Westby, Katie M; Fritzen, Charissa; Paulsen, Dave; Poindexter, Stephanie; Moncayo, Abelardo C

    2015-09-01

    La Crosse virus (LACV) is a mosquito-borne virus and a major cause of pediatric encephalitis in the USA. La Crosse virus emerged in Tennessee and other states in the Appalachian region in 1997. We investigated LACV infection rates and seasonal abundances of the native mosquito vector, Aedes triseriatus, and 2 recently introduced mosquito species, Ae. albopictus and Ae. japonicus, in an emerging disease focus in Tennessee. Mosquitoes were collected using multiple trapping methods specific for Aedes mosquitoes at recent human case sites. Mosquito pools were tested via reverse transcriptase-polymerase chain reaction (RT-PCR) of the S segment to detect multiple Bunyamwera and California serogroup viruses, including LACV, as well as real-time RT-PCR of the M segment. A total of 54 mosquito pools were positive, including wild-caught adult females and laboratory-reared adults, demonstrating transovarial transmission in all 3 species. Maximum likelihood estimates (per 1,000 mosquitoes) were 2.72 for Ae. triseriatus, 3.01 for Ae. albopictus, and 0.63 for Ae. japonicus. We conclude that Ae. triseriatus and Ae. albopictus are important LACV vectors and that Ae. japonicus also may be involved in virus maintenance and transmission.

  15. Field evaluation of the Off! Clip-on Mosquito Repellent (metofluthrin) against Aedes albopictus and Aedes taeniorhynchus (Diptera: Culicidae) in northeastern Florida.

    PubMed

    Xue, Rui-De; Qualls, Whitney A; Smith, Michael L; Gaines, Marcia K; Weaver, James H; Debboun, Mustapha

    2012-05-01

    Repellent efficacy of the Off! Clip-on Mosquito Repellent device (S. C. Johnson and Son, Inc., Racine, WI) containing Metofluthrin was evaluated on six human volunteers against the container-breeding mosquito Aedes albopictus (Skuse) and the salt marsh mosquito Aedes taeniorhynchus (Wiedemann) at two field locations in northeastern Florida. The device repelled mosquitoes by releasing a vaporized form of the pyrethroid insecticide metofluthrin ([AI] 31.2%) and provided 70% protection from Ae. albopictus bites for > 3 h. For the second field trial, a repellent device that was used in the first trial was tested after being open for >1 wk. This device provided 79% protection from Ae. taeniorhynchus bites for 3 h. Our field results showed that the repellent device was 70 and 79% effective at repelling Ae. albopictus and Ae. taeniorhynchus from human test subjects in both field locations in northeastern Florida.

  16. Ultra low concentration deltamethrin loaded patch development and evaluation of its repellency against dengue vector Aedes (S) albopictus

    PubMed Central

    2013-01-01

    Background Mosquito repellents and emanators confer protection against mosquito bites through spatial action of emanated vapours which are released into the adjoining environment. Synthetic insecticides released into the environment in ultra low volume vapour phase deter the mosquitoes from biting humans in a protected space. Methods Formulation patches were prepared using the solvent evaporation method over a backing membrane and using Dibutylphthalate (DBT) as a plasticizer. The effect of formulation variables on the deltamethrin release from the patch matrices were studied under accelerated conditions, whereas, HPLC was used for quantitative estimation of deltamethrin. The prepared patch formulations were subjected to physicochemical studies, such as, deltamethrin content, thickness, weight variation, percent moisture content, moisture uptake, surface area and surface pH determination. Deltamethrin-polymer interaction and compatibility was ascertained using DSC and FT-IR, while surface morphology and deltamethrin distribution in the patch were studied using SEM technique. Repellent activity of the patch formulations was evaluated against Ae. albopictus mosquitoes. Results Blends of polymeric combinations of polyvinylpyrrolidone (PVP) and ethyl-cellulose (EC) with admixture of deltamethrin provided prolonged repellent activity against Ae. albopictus mosquitoes. Physicochemical characterisation indicated the suitability of deltamethrin patch formulation with the polymeric combinations of PVP and EC. Patches were very effective against laboratory reared Ae. albopictus mosquitoes. No significant difference was observed between the performance of test patches and commercially available repellent cream Mosqshield®. Conclusion Deltamethrin loaded patches provided effective repellency against Ae. albopictus mosquitoes. The study emphasised that deltamethrin released to the environment in low concentration could be an excellent spatial repellent against hematophagous

  17. Breeding of Aedes aegypti (L.) and Aedes albopictus (Skuse) in urban housing of Sibu town, Sarawak.

    PubMed

    Seng, C M; Jute, N

    1994-09-01

    An Aedes survey using various larval survey methods was conducted in 12 urban housing areas and 29 vacant lands in Sibu town proper. Aedes albopictus larvae were found in all areas surveyed while Aedes aegypti larvae were present in 10 localities and 4 vacant lands. There were no significant difference in the house index, breteau and larval density index of these two Aedes (Stegomyia) species from the survey areas. The proportion of containers positive with Ae. aegypti and Ae. albopictus in area outside the house compound and near the house fencing were 3.2 times higher than outdoor compound. The indoor/outdoor breeding ratio for Ae. aegypti alone is 1.6:1. The most preferred breeding habitats outdoor were plastic cups and used tires while indoor habitats were ant traps and flower vases. In the vacant lands, the average number of larvae per containers was significantly higher than in houses and over 51% of the containers inspected were positive. Shared breeding between Ae. aegypti and Ae. albopictus larvae accounted for 9% in house surveys and 4.5% in vacant land survey. The use of various methods in Aedes larval survey may provide essential information in the study of vector epidemiology in dengue and dengue hemorrhagic fever transmission.

  18. Differential Larval Toxicity and Oviposition Altering Activity of Some Indigenous Plant Extracts against Dengue and Chikungunya Vector Aedes albopictus.

    PubMed

    Yadav, Ruchi; Tyagi, Varun; Tikar, Sachin N; Sharma, Ajay K; Mendki, Murlidhar J; Jain, Ashok K; Sukumaran, Devanathan

    2014-12-01

    Mosquitoes are well known as vectors of several disease causing pathogens. The extensive use of synthetic insecticides in the mosquito control strategies resulted to the development of pesticide resistance and fostered environmental deterioration. Hence in recent years plants become alternative source of mosquito control agents. The present study assessed the larvicidal and oviposition altering activity of six different plants species-Alstonia scholaris, Callistemon viminalis, Hyptis suaveolens, Malvastrum coromandelianum, Prosopis juliflora, Vernonia cinerea against Aedes albopictus mosquito in laboratory. Leaf extracts of all the six plants species in five different solvents of various polarities were used in the range of 20-400ppm for larval bioassay and 50,100 and 200ppm for cage bioassay (for the study of oviposition behavior) against Ae. albopictus. The larval mortality data were recorded after 24 h and subjected to Probit analysis to determine the lethal concentrations (LC50), while OAI (Oviposition activity index) was calculated for oviposition altering activity of the plant extracts. Vernonia cinerea extract in acetone and C. viminalis extract in isopropanol were highly effective against Aedes albopictus larvae with LC50 value 64.57, 71.34ppm respectively. Acetone extract of P. juliflora found to be strong oviposition-deterrent which inhibited >2 fold egg laying (OAI-0.466) at 100ppm. Vernonia cinerea and C. viminallis leaf extracts have the potential to be used as larvicide and P. juliflora as an oviposition-deterrent for the control of Ae. albopictus mosquito.

  19. Differential Larval Toxicity and Oviposition Altering Activity of Some Indigenous Plant Extracts against Dengue and Chikungunya Vector Aedes albopictus

    PubMed Central

    Yadav, Ruchi; Tyagi, Varun; Tikar, Sachin N; Sharma, Ajay K; Mendki, Murlidhar J; Jain, Ashok K; Sukumaran, Devanathan

    2014-01-01

    Background: Mosquitoes are well known as vectors of several disease causing pathogens. The extensive use of synthetic insecticides in the mosquito control strategies resulted to the development of pesticide resistance and fostered environmental deterioration. Hence in recent years plants become alternative source of mosquito control agents. The present study assessed the larvicidal and oviposition altering activity of six different plants species-Alstonia scholaris, Callistemon viminalis, Hyptis suaveolens, Malvastrum coromandelianum, Prosopis juliflora, Vernonia cinerea against Aedes albopictus mosquito in laboratory. Methods: Leaf extracts of all the six plants species in five different solvents of various polarities were used in the range of 20–400ppm for larval bioassay and 50,100 and 200ppm for cage bioassay (for the study of oviposition behavior) against Ae. albopictus. The larval mortality data were recorded after 24 h and subjected to Probit analysis to determine the lethal concentrations (LC50), while OAI (Oviposition activity index) was calculated for oviposition altering activity of the plant extracts. Results: Vernonia cinerea extract in acetone and C. viminalis extract in isopropanol were highly effective against Aedes albopictus larvae with LC50 value 64.57, 71.34ppm respectively. Acetone extract of P. juliflora found to be strong oviposition-deterrent which inhibited >2 fold egg laying (OAI-0.466) at 100ppm. Conclusion: Vernonia cinerea and C. viminallis leaf extracts have the potential to be used as larvicide and P. juliflora as an oviposition-deterrent for the control of Ae. albopictus mosquito. PMID:26114131

  20. Natural transmission of dengue virus serotype 3 by Aedes albopictus (Skuse) during an outbreak in Havelock Island: Entomological characteristics.

    PubMed

    Sivan, Arun; Shriram, A N; Sugunan, A P; Anwesh, Maile; Muruganandam, N; Kartik, C; Vijayachari, P

    2016-04-01

    From May to June 2014, an outbreak of dengue virus (DENV) illness occurred in the Havelock Island, South Andaman. Entomological investigations were undertaken during the peak of the outbreak, from 26th May-4th June, to identify the primary vector(s) involved in the transmission so that appropriate public health measures could be implemented. Adult mosquitoes were collected by BG-Sentinel traps in houses and neighborhoods of clinically ill patients. Water holding containers were inspected for the presence of mosquito larvae and pupae. Adult mosquitoes were analyzed by RT-PCR for the presence of nucleic acids of DENV and CHIKV. A total of 498 mosquitoes were collected and processed in 27 pools. The species composition comprised of 58.3% Aedes albopictus, 7.5% Aedes aegypti and 4.2% Aedes edwardsi and 3.1% constituted others. Two A. albopictus pools were found to be positive for DENV RNA. Sequencing of the RT PCR 511 base pair amplicon positive samples showed homology with DENV-3, suggesting that serotype-3 was responsible for the outbreak and A. albopictus was the primary vector responsible. This was supported by high container (10.1%), premise (25.4%) and Breteau (27.9) indices, with miscellaneous receptacles (2.4%), tree holes (1.2%) and discarded tires (1.2%) registering relatively higher container indices. This is the first report of detection of DENV in A. albopictus from Andaman and Nicobar Islands. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. 77 FR 5489 - Identification of Human Cell Lines Project

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-03

    ...-01] Identification of Human Cell Lines Project AGENCY: National Institute of Standards and Technology... cell line samples as part of the Identification of Human Cell Lines Project. All data and corresponding... cell lines accepted on the NIST Applied Genetics Group Web site at http://www.nist.gov/mml/biochemical...

  2. Interspecific competition of a new invasive mosquito, Culex coronator, and two container mosquitoes, Aedes albopictus and Cx. quinquefasciatus (Diptera: Culicidae), across different detritus environments.

    PubMed

    Yee, D A; Skiff, J F

    2014-01-01

    The mosquito Culex coronator (Dyar and Knab) (Diptera: Culicidae) has undergone rapid range expansion in the United States since 2003, with its historical distribution in the southwest expanding eastward to the Atlantic coast. Although Cx. coronator nominally use small natural aquatic habitats for development, the use of containers (e.g., tires) makes it potentially important as container invasive. To determine the potential ecological effects of Cx. coronator on resident container species, we conducted a laboratory experiment to assess its competitive ability with two common tire-inhabiting species, Aedes albopictus (Skuse) and Culex quinquefasciatus (Say) (Diptera: Culicidae). Larvae were reared under a factorial design with each species alone and in combination (Cx. coronator + Ae. albopictus, Cx. coronator + Cx. quinquefasciatus) across three different resource environments (leaf detritus only, animal detritus only, animal + leaf). Mosquito performance (survival, adult male and female mass, and development time) was measured for each species across treatments. Female Cx. coronator developed slowest when grown with Ae. albopictus, or when grown with leaves only regardless of species combinations; similar patterns emerged for males although species effects were restricted to mass. Few differences were evident in performance for male and female Cx. coronator across detritus environments when grown with Cx. quinquefasciatus. Cx. quinquefasciatus did not vary in mass or development time in the presence of Cx. coronator compared with when grown alone. Ae. albopictus female mass was 15% lower in the presence of Cx. coronator. Survival of Cx. coronator was highest in animal and leaf detritus containers, although survival was generally lower when larvae were grown with Ae. albopictus. These findings suggest that the performance of Cx. coronator is similar to that of Cx. quinquefasciatus but it suffers in the presence of Ae. albopictus under some resource environments.

  3. Macrocyclops albidus (Copepoda: cyclopidae) for the Biocontrol of Aedes albopictus and Culex pipiens in Italy.

    PubMed

    Veronesi, Rodolfo; Carrieri, Marco; Maccagnani, Bettina; Maini, Stefano; Bellini, Romeo

    2015-03-01

    The aim of our study was to assess the potential of Macrocyclops albidus as a biological control agent against the 1st and 2nd instars of Culex pipiens and Aedes albopictus. Under laboratory conditions of prey saturation (50 1st instars/copepod), an average of 58.98% of Cx. pipiens and 54.99% of Ae. albopictus larvae were killed by 1 copepod in 24 h. Trials run in big drums containing 200 liters of water showed that the M. albidus population, inoculated in April, efficiently controlled the mosquito population for the entire season. The predator was particularly effective against Ae. albopictus, as only 2 larvae of this species were found in the treated drums, compared to 814 larvae in untreated control drums throughout the study period. No difference was observed in the control efficacy between the 2 initial densities of copepods used. The reduction in Ae. albopictus density in the drums with 100 and 500 M. albidus with respect to the control drums was 99.90 ± 0.35% and 100.0 ± 0.0%, respectively. For Cx. pipiens, the reduction in density was 88.69 ± 13.51% and 84.65% in drums inoculated with 100 and 500 copepods, respectively. Macrocyclops albidus populations survived through the winter and continued to keep the mosquito population under control during the 2008 season. The M. albidus population developed very well both in drums placed in sunny and shaded areas and proved to be tolerant to both high (summer) and low (winter) temperatures. Trials performed on M. albidus survival in catch basins showed that after a few weeks, the copepod population dramatically decreased and subsequently disappeared. The main problem for copepod survival in catch basins seemed to be the low oxygen tension and accumulation of toxic substances, rather than copepods being flushed out in heavy rainfall episodes. During the period when copepods were present, they maintained the mosquito population under control; their partial disappearance from the catch basins, however, would

  4. Spread of the Tiger: Global Risk of Invasion by the Mosquito Aedes albopictus

    PubMed Central

    BENEDICT, MARK Q.; LEVINE, REBECCA S.; HAWLEY, WILLIAM A.; LOUNIBOS, L. PHILIP

    2008-01-01

    Aedes albopictus, commonly known as the Asian tiger mosquito, is currently the most invasive mosquito in the world. It is of medical importance due to its aggressive daytime human-biting behavior and ability to vector many viruses, including dengue, LaCrosse, and West Nile. Invasions into new areas of its potential range are often initiated through the transportation of eggs via the international trade in used tires. We use a genetic algorithm, Genetic Algorithm for Rule Set Production (GARP), to determine the ecological niche of Ae. albopictus and predict a global ecological risk map for the continued spread of the species. We combine this analysis with risk due to importation of tires from infested countries and their proximity to countries that have already been invaded to develop a list of countries most at risk for future introductions and establishments. Methods used here have potential for predicting risks of future invasions of vectors or pathogens. PMID:17417960

  5. Invasiveness of Aedes aegypti and Aedes albopictus and Vectorial Capacity for Chikungunya Virus

    PubMed Central

    Lounibos, Leon Philip; Kramer, Laura D.

    2016-01-01

    In this review, we highlight biological characteristics of Aedes aegypti and Aedes albopictus, 2 invasive mosquito species and primary vectors of chikungunya virus (CHIKV), that set the tone of these species' invasiveness, vector competence, and vectorial capacity (VC). The invasiveness of both species, as well as their public health threats as vectors, is enhanced by preference for human blood. Vector competence, characterized by the efficiency of an ingested arbovirus to replicate and become infectious in the mosquito, depends largely on vector and virus genetics, and most A. aegypti and A. albopictus populations thus far tested confer vector competence for CHIKV. VC, an entomological analog of the pathogen's basic reproductive rate (R0), is epidemiologically more important than vector competence but less frequently measured, owing to challenges in obtaining valid estimates of parameters such as vector survivorship and host feeding rates. Understanding the complexities of these factors will be pivotal in curbing CHIKV transmission. PMID:27920173

  6. How does the dengue vector mosquito Aedes albopictus respond to global warming?

    PubMed

    Jia, Pengfei; Chen, Xiang; Chen, Jin; Lu, Liang; Liu, Qiyong; Tan, Xiaoyue

    2017-03-11

    Global warming has a marked influence on the life cycle of epidemic vectors as well as their interactions with human beings. The Aedes albopictus mosquito as the vector of dengue fever surged exponentially in the last decade, raising ecological and epistemological concerns of how climate change altered its growth rate and population dynamics. As the global warming pattern is considerably uneven across four seasons, with a confirmed stronger effect in winter, an emerging need arises as to exploring how the seasonal warming effects influence the annual development of Ae. albopictus. The model consolidates a 35-year climate dataset and designs fifteen warming patterns that increase the temperature of selected seasons. Based on a recently developed mechanistic population model of Ae. albopictus, the model simulates the thermal reaction of blood-fed adults by systematically increasing the temperature from 0.5 to 5 °C at an interval of 0.5 °C in each warming pattern. The results show the warming effects are different across seasons. The warming effects in spring and winter facilitate the development of the species by shortening the diapause period. The warming effect in summer is primarily negative by inhibiting mosquito development. The warming effect in autumn is considerably mixed. However, these warming effects cannot carry over to the following year, possibly due to the fact that under the extreme weather in winter the mosquito fully ceases from development and survives in terms of diapause eggs. As the historical pattern of global warming manifests seasonal fluctuations, this study provides corroborating and previously ignored evidence of how such seasonality affects the mosquito development. Understanding this short-term temperature-driven mechanism as one chain of the transmission events is critical to refining the thermal reaction norms of the epidemic vector under global warming as well as developing effective mosquito prevention and control strategies.

  7. Vertical Transmission of Zika Virus by Aedes aegypti and Ae. albopictus Mosquitoes.

    PubMed

    Ciota, Alexander T; Bialosuknia, Sean M; Ehrbar, Dylan J; Kramer, Laura D

    2017-05-01

    To determine the potential role of vertical transmission in Zika virus expansion, we evaluated larval pools of perorally infected Aedes aegypti and Ae. albopictus adult female mosquitoes; ≈1/84 larvae tested were Zika virus-positive; and rates varied among mosquito populations. Thus, vertical transmission may play a role in Zika virus spread and maintenance.

  8. Feeder-cell-independent culture of the pig-embryonic-stem-cell-derived exocrine pancreatic cell line, PICM-31

    USDA-ARS?s Scientific Manuscript database

    The adaptation to feeder-independent growth of a pig embryonic stem cell-derived pancreatic cell line is described. The parental PICM-31 cell line, previously characterized as an exocrine pancreas cell line, was colony-cloned two times in succession resulting in the subclonal cell line, PICM-31A1. P...

  9. Modeling the Environmental Suitability for Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus (Diptera: Culicidae) in the Contiguous United States.

    PubMed

    Johnson, Tammi L; Haque, Ubydul; Monaghan, Andrew J; Eisen, Lars; Hahn, Micah B; Hayden, Mary H; Savage, Harry M; McAllister, Janet; Mutebi, John-Paul; Eisen, Rebecca J

    2017-11-07

    The mosquitoes Aedes (Stegomyia) aegypti (L.)(Diptera:Culicidae) and Ae. (Stegomyia) albopictus (Skuse) (Diptera:Culicidae) transmit dengue, chikungunya, and Zika viruses and represent a growing public health threat in parts of the United States where they are established. To complement existing mosquito presence records based on discontinuous, non-systematic surveillance efforts, we developed county-scale environmental suitability maps for both species using maximum entropy modeling to fit climatic variables to county presence records from 1960-2016 in the contiguous United States. The predictive models for Ae. aegypti and Ae. albopictus had an overall accuracy of 0.84 and 0.85, respectively. Cumulative growing degree days (GDDs) during the winter months, an indicator of overall warmth, was the most important predictive variable for both species and was positively associated with environmental suitability. The number (percentage) of counties classified as environmentally suitable, based on models with 90 or 99% sensitivity, ranged from 1,443 (46%) to 2,209 (71%) for Ae. aegypti and from 1,726 (55%) to 2,329 (75%) for Ae. albopictus. Increasing model sensitivity results in more counties classified as suitable, at least for summer survival, from which there are no mosquito records. We anticipate that Ae. aegypti and Ae. albopictus will be found more commonly in counties classified as suitable based on the lower 90% sensitivity threshold compared with the higher 99% threshold. Counties predicted suitable with 90% sensitivity should therefore be a top priority for expanded mosquito surveillance efforts while still keeping in mind that Ae. aegypti and Ae. albopictus may be introduced, via accidental transport of eggs or immatures, and potentially proliferate during the warmest part of the year anywhere within the geographic areas delineated by the 99% sensitivity model. Published by Oxford University Press on behalf of Entomological Society of America 2017. This work

  10. Effects of infectious virus dose and bloodmeal delivery method on susceptibility of Aedes aegypti and Aedes albopictus to chikungunya virus.

    PubMed

    Pesko, Kendra; Westbrook, Catherine J; Mores, Christopher N; Lounibos, L Philip; Reiskind, Michael H

    2009-03-01

    Chikungunya virus (CHIKV) is an arbovirus (genus Alphavirus, family Togaviridae) that has recently caused disease outbreaks in the Indian Ocean basin and southern Europe. These outbreaks could be associated with a possible shift in primary vector from Aedes aegypti to Ae. albopictus. To evaluate vector competence differences in possible CHIKV vectors, we evaluated the dose-dependant susceptibility of Florida strains of Ae. albopictus and Ae. aegypti for infection with a La Réunion island strain of CHIKV. Pledget and water-jacketed membrane feeding systems were also evaluated. We show that both Aedes spp. were susceptible to the highest CHIKV doses, whereas only Ae. albopictus developed disseminated infections after exposure to the two lowest doses. Infection rates for both mosquito species were significantly affected by the bloodmeal delivery method used. This information is important in assessing risk of an outbreak of imported CHIKV in the United States, in determining differences in vectorial capacity of these two vector species, and in evaluating arbovirus delivery methods in the laboratory.

  11. The tale of two buckets and associated containers: impact on aedes albopictus oviposition

    USDA-ARS?s Scientific Manuscript database

    Aedes albopictus is an invasive species. Its oviposition behavior is the subject of several projects in our research unit. The main emphasis of this presentation is a study which utilizes two five gallon buckets, one heated and one with ambient temperature. The heat is provided by an aquarium hea...

  12. Aedes albopictus (Skuse, 1894) infected with the American-Asian genotype of dengue type 2 virus in Medellín suggests its possible role as vector of dengue fever in Colombia.

    PubMed

    Gómez-Palacio, Andrés; Suaza-Vasco, Juan; Castaño, Sandra; Triana, Omar; Uribe, Sandra

    2017-03-29

    Aedes aegypti and Ae. albopictus are recognized vectors of dengue, yellow fever, chikungunya and Zika arboviruses in several countries worldwide. In Colombia, Ae. albopictus geographical distribution has increased to include highly populated cities such as Cali and Medellín. Although this species has been frequently found in urban and semi-urban zones in the country, its role as vector of the dengue fever is poorly known. To identify the presence of Ae. albopictus specimens naturally infected with dengue virus collected in Medellín. Insects were collected in the Universidad Nacional de Colombia campus in Medellín. Individuals were classified as Ae. albopictus and confirmed by DNA barcode region analysis. Mosquitoes were processed for dengue virus identification, and a fragment of the NS3 gen was sequenced and compared with DENV-2 genotypes reported in the literature. Sequence analysis of COI indicated Ae. albopictus individuals were similar to those recently reported in Colombia, and genetically close to those from other regions worldwide. Among the pools tested one was positive for DENV-2, and the NS3 analysis indicated it belonged to the Asian-American clade. We report the presence Ae. albopictus naturally infected with the Asian-American genotype of DENV-2 in Colombia. The presence of Ae. albopictus specimens carrying the most common genotype infecting humans in a highly populated city such as Medellín indicates its potential role as dengue vector in Colombia and highlights the relevance of including it in current vector surveillance strategies.

  13. 3-Bromopyruvate induces necrotic cell death in sensitive melanoma cell lines.

    PubMed

    Qin, J-Z; Xin, H; Nickoloff, B J

    2010-05-28

    Clinicians successfully utilize high uptake of radiolabeled glucose via PET scanning to localize metastases in melanoma patients. To take advantage of this altered metabolome, 3-bromopyruvate (BrPA) was used to overcome the notorious resistance of melanoma to cell death. Using four melanoma cell lines, BrPA triggered caspase independent necrosis in two lines, whilst the other two lines were resistant to killing. Mechanistically, sensitive cells differed from resistant cells by; constitutively lower levels of glutathione, reduction of glutathione by BrPA only in sensitive cells; increased superoxide anion reactive oxygen species, loss of outer mitochondrial membrane permeability, and rapid ATP depletion. Sensitive cell killing was blocked by N-acetylcysteine or glutathione. When glutathione levels were reduced in resistant cell lines, they became sensitive to killing by BrPA. Taken together, these results identify a metabolic-based Achilles' heel in melanoma cells to be exploited by use of BrPA. Future pre-clinical and clinical trials are warranted to translate these results into improved patient care for individuals suffering from metastatic melanoma. Copyright (c) 2010 Elsevier Inc. All rights reserved.

  14. Tuft (caveolated) cells in two human colon carcinoma cell lines.

    PubMed

    Barkla, D H; Whitehead, R H; Foster, H; Tutton, P J

    1988-09-01

    The presence of an unusual cell type in two human colon carcinoma cell lines is reported. The cells show the same morphology as "tuft" (caveolated) cells present in normal gastrointestinal epithelium. Tuft cells were seen in cell line LIM 1863 growing in vitro and in human colon carcinoma cell line LIM 2210 growing as subcutaneous solid tumour xenografts in nude mice. Characteristic morphologic features of tuft cells included a wide base, narrow apex and a tuft of long microvilli projecting from the apical surface. The microvilli are attached by a core of long microfilaments passing deep into the apical cytoplasm. Between the microvilli are parallel arrays of vesicles (caveoli) containing flocculent material. Two different but not mutually exclusive explanations for the presence of tuft cells are proposed. The first explanation is that tuft cells came from the resected tumour and have survived by mitotic division during subsequent passages. The second explanation suggests that tuft cells are the progeny of undifferentiated tumour cells. Descriptions of tuft cells in colon carcinomas are uncommon and possible reasons for this are presented. The morphology of tuft cells is consistent with that of a highly differentiated cell specialised for absorption, and these new models provide an opportunity to further investigate the structure and function of tuft cells.

  15. Leukemia-lymphoma cell lines as model systems for hematopoietic research.

    PubMed

    Drexler, Hans G; MacLeod, Roderick A F

    2003-01-01

    Continuous human leukemia-lymphoma (LL) cell lines comprise a rich self-renewing resource of accessible and manipulable living cells which has illuminated the pathophysiology of hematopoietic tumors as well as basic cell biology. The major key advantages of continuous cell lines are the unlimited supply and worldwide availability of identical cell material and their cryopreservation. LL cell lines are characterized generally by monoclonal origin and differentiation arrest, sustained proliferation in vitro with preservation of most cellular features, and specific genetic alterations. The most practical classification of LL cell lines assigns them to one of the physiologically occurring cell lineages, based on their immunophenotype, genotype and functional features. Truly malignant cell lines may be distinguished from Epstein-Barr virus (EBV)-immortalized normal cells, using various operational and conceptual parameters. The characterization and publication of new LL cell lines provides important and informative core data which, by opening new avenues for investigation, have become ubiquitous powerful research tools that are available to every investigator by reference cell repositories. There is a need in the scientific community for clean and authenticated LL cell lines to which every scientist has access as offered by these institutionalized public cell line banks. A list of the most useful, robust and freely available reference cell lines is proposed in this review. Clearly, studies of LL cell lines have provided seminal insights into the biology of hematopoietic neoplasia.

  16. Continuous human cell lines and method of making same

    DOEpatents

    Stampfer, Martha R.

    1989-01-01

    Substantially genetically stable continuous human cell lines derived from normal human mammary epithelial cells (HMEC) and processes for making and using the same. In a preferred embodiment, the cell lines are derived by treating normal human mammary epithelial tissue with a chemical carcinogen such as benzo[a]pyrene. The novel cell lines serve as useful substrates for elucidating the potential effects of a number of toxins, carcinogens and mutagens as well as of the addition of exogenous genetic material. The autogenic parent cells from which the cell lines are derived serve as convenient control samples for testing. The cell lines are not neoplastically transformed, although they have acquired several properties which distinguish them from their normal progenitors.

  17. Continuous human cell lines and method of making same

    DOEpatents

    Stampfer, M.R.

    1985-07-01

    Substantially genetically stable continuous human cell lines derived from normal human mammary epithelial cells (HMEC) and processes for making and using the same. In a preferred embodiment, the cell lines are derived by treating normal human mammary epithelial tissue with a chemical carcinogen such as benzo(a)pyrene. The novel cell lines serve as useful substrates for elucidating the potential effects of a number of toxins, carcinogens and mutagens as well as of the addition of exogenous genetic material. The autogenic parent cells from which the cell lines are derived serve as convenient control samples for testing. The cell lines are not neoplastically transformed, although they have acquired several properties which distinguish them from their normal progenitors. 2 tabs.

  18. Effects of curcumin on stem-like cells in human esophageal squamous carcinoma cell lines.

    PubMed

    Almanaa, Taghreed N; Geusz, Michael E; Jamasbi, Roudabeh J

    2012-10-24

    Many cancers contain cell subpopulations that display characteristics of stem cells. Because these cancer stem cells (CSCs) appear to provide resistance to chemo-radiation therapy, development of therapeutic agents that target CSCs is essential. Curcumin is a phytochemical agent that is currently used in clinical trials to test its effectiveness against cancer. However, the effect of curcumin on CSCs is not well established. The current study evaluated curcumin-induced cell death in six cancer cell lines derived from human esophageal squamous cell carcinomas. Moreover, these cell lines and the ones established from cells that survived curcumin treatments were characterized. Cell loss was assayed after TE-1, TE-8, KY-5, KY-10, YES-1, and YES-2 cells were exposed to 20-80 μM curcumin for 30 hrs. Cell lines surviving 40 or 60 μM curcumin were established from these six original lines. The stem cell markers aldehyde dehydrogenase-1A1 (ALDH1A1) and CD44 as well as NF-κB were used to compare CSC-like subpopulations within and among the original lines as well as the curcumin-surviving lines. YES-2 was tested for tumorsphere-forming capabilities. Finally, the surviving lines were treated with 40 and 60 μM curcumin to determine whether their sensitivity was different from the original lines. The cell loss after curcumin treatment increased in a dose-dependent manner in all cell lines. The percentage of cells remaining after 60 μM curcumin treatment varied from 10.9% to 36.3% across the six lines. The cell lines were heterogeneous with respect to ALDH1A1, NF-κB and CD44 expression. KY-5 and YES-1 were the least sensitive and had the highest number of stem-like cells whereas TE-1 had the lowest. The curcumin-surviving lines showed a significant loss in the high staining ALDH1A1 and CD44 cell populations. Tumorspheres formed from YES-2 but were small and rare in the YES-2 surviving line. The curcumin-surviving lines showed a small but significant decrease in sensitivity

  19. Synthesis of some novel phosphorylated and thiophosphorylated benzimidazoles and benzothiazoles and their evaluation for larvicidal potential to Aedes albopictus and Culex quinquefasciatus.

    PubMed

    Bandyopadhyay, Prabal; Sathe, Manisha; Tikar, Sachin N; Yadav, Ruchi; Sharma, Pratibha; Kumar, Ashok; Kaushik, M P

    2014-07-01

    Series of benzimidazole and benzothiazole linked phosphoramidates and phosphoramidothioates (5a-j) and benzimidazole linked phenylphosphoramidates and phenylphosphoramidothioates (10a-e) were synthesized. The title compounds were preliminary screened for mosquito larvicidal properties against Aedes albopictus and Culex quinquefasciatus at different concentration from 40 to 5 mg/L. Among the screened compounds three compounds revealed potential larvicidal effects with 100% mortality in the order of 10e>5j>5e. Compound 10e was found to be the most toxic compound to Ae. albopictus and Cx. quinquefasciatus. The LC50 of 10e against Ae. albopictus was found to be 6.42 and 5.25 mg/L at 24 and 48 h, respectively, whereas it was 7.01 and 3.88 mg/L, respectively in Cx. quinquefasciatus. Temephos was used as positive control. Copyright © 2014 Elsevier Ltd. All rights reserved.

  20. Influence of multiple blood meals on gonotrophic dissociation and fecundity in Aedes albopictus

    USDA-ARS?s Scientific Manuscript database

    Female Aedes albopictus blood fed on guinea pig and human hosts produced significantly (P < 0.05) higher number of eggs (80 and 82/female, respectively) than females fed on chicken (67 eggs/female). Fecundity in mosquitoes that took a double blood meal (chicken and guinea pig), a triple blood meal ...

  1. Vertical transmission of Key West dengue-1 virus by Aedes aegypti and Aedes albopictus (Diptera: Culicidae) mosquitoes from Florida.

    PubMed

    Buckner, Eva A; Alto, Barry W; Lounibos, L Philip

    2013-11-01

    Following the 2009 and 2010 dengue-1 (DENV-1) outbreaks in Key West, FL, we used Florida Aedes aegypti (L.) mosquitoes and DENV-1 isolated from Key West in 2010 to test the hypothesis that if the 2009 and 2010 DENV-1 genome sequences are similar, then vertical transmission of DENV-1 from infected Ae. aegypti female mosquitoes to their eggs could have served as an interepidemic reservoir between outbreaks. We also investigated the ability of Florida Aedes albopictus (Skuse) mosquitoes to vertically transmit DENV-1. In addition, we determined the rates of infection and dissemination of these Florida mosquito species for DENV-1 and the effect of DENV-1 infection on oviposition success and number of mosquito eggs laid by females. Vertical transmission of DENV-1 was documented, with rates of 11.11% (2 out of 18) for Ae. albopictus and 8.33% (3 out of 36) for Ae. aegypti. Approximately 93% (111 out of 119) of Ae. aegypti that fed on DENV-1 in blood became infected, and 80% (89 out of 111) of infections were disseminated. Similarly, 93% of Ae. albopictus became infected (53 out of 57), and 85% (45 out of 53) of infections were disseminated. No significant differences were detected in numbers of eggs laid by either species after imbibing DENV-1 in blood, suggesting little cost of infection on number of eggs laid. Our results demonstrate that Florida Ae. aegypti and Ae. albopictus mosquitoes are competent vectors for DENV-1, whose maintenance between the 2009 and 2010 Key West outbreaks may have been facilitated by vertical transmission.

  2. Replication of Heliothis virescens ascovirus in insect cell lines.

    PubMed

    Asgari, S

    2006-09-01

    Ascoviruses (AVs) infect larvae of various insect pests belonging to the family Noctuidae. The result of AV infection in the hosts is cleavage of infected cells into vesicles, a unique feature of AV infection. Since insect cell lines facilitate the study of virus life cycles, attempts were made to analyze Heliothis virescens AV (HvAV3e) infection in several cell lines and compare cell pathology to larval infection. In this study, replication and cytopathological effects of HvAV3e on four different cell lines were investigated. HvAV3e replication was confirmed in three noctuid cell lines from Spodoptera frugiperda (Sf9) and Helicoverpa zea (BCIRL-Hz-AM1 and FB33). However, the virus did not replicate in the non-noctuid insect cell line from Pieris rapae (Pieridae). Despite replication of the virus in the three permissive cell lines, the cytopathological effects of the virus were significantly different from that of larval infection.

  3. 3-Bromopyruvate induces necrotic cell death in sensitive melanoma cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Qin, J.-Z.; Xin, H.; Nickoloff, B.J., E-mail: bnickol@lumc.edu

    2010-05-28

    Clinicians successfully utilize high uptake of radiolabeled glucose via PET scanning to localize metastases in melanoma patients. To take advantage of this altered metabolome, 3-bromopyruvate (BrPA) was used to overcome the notorious resistance of melanoma to cell death. Using four melanoma cell lines, BrPA triggered caspase independent necrosis in two lines, whilst the other two lines were resistant to killing. Mechanistically, sensitive cells differed from resistant cells by; constitutively lower levels of glutathione, reduction of glutathione by BrPA only in sensitive cells; increased superoxide anion reactive oxygen species, loss of outer mitochondrial membrane permeability, and rapid ATP depletion. Sensitive cellmore » killing was blocked by N-acetylcysteine or glutathione. When glutathione levels were reduced in resistant cell lines, they became sensitive to killing by BrPA. Taken together, these results identify a metabolic-based Achilles' heel in melanoma cells to be exploited by use of BrPA. Future pre-clinical and clinical trials are warranted to translate these results into improved patient care for individuals suffering from metastatic melanoma.« less

  4. Variability of human pluripotent stem cell lines.

    PubMed

    Ortmann, Daniel; Vallier, Ludovic

    2017-10-01

    Human pluripotent stem cells derived from embryos (human Embryonic Stem Cells or hESCs) or generated by direct reprogramming of somatic cells (human Induced Pluripotent Stem Cells or hiPSCs) can proliferate almost indefinitely in vitro while maintaining the capacity to differentiate into a broad diversity of cell types. These two properties (self-renewal and pluripotency) confers human pluripotent stem cells a unique interest for clinical applications since they could allow the production of infinite quantities of cells for disease modelling, drug screening and cell based therapy. However, recent studies have clearly established that human pluripotent stem cell lines can display variable capacity to differentiate into specific lineages. Consequently, the development of universal protocols of differentiation which could work efficiently with any human pluripotent cell line is complicated substantially. As a consequence, each protocol needs to be adapted to every cell line thereby limiting large scale applications and precluding personalised therapies. Here, we summarise our knowledge concerning the origin of this variability and describe potential solutions currently available to bypass this major challenge. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Continuous hematopoietic cell lines as model systems for leukemia-lymphoma research.

    PubMed

    Drexler, H G; Matsuo, A Y; MacLeod, R A

    2000-11-01

    Along with other improvements, the advent of continuous human leukemia-lymphoma (LL) cell lines as a rich resource of abundant, accessible and manipulable living cells has contributed significantly to a better understanding of the pathophysiology of hematopoietic tumors. The first LL cell lines, Burkitt's lymphoma-derived lines, were established in 1963. Since then, more than 1000 cell lines have been described, although not all of them in full detail. The major advantages of continuous cell lines is the unlimited supply and worldwide availability of identical cell material, and the infinite viable storability in liquid nitrogen. LL cell lines are characterized generally by monoclonal origin and differentiation arrest, sustained proliferation in vitro under preservation of most cellular features, and specific genetic alterations. The most practical classification of LL cell lines assigns them to one of the physiologically occurring cell lineages, based on their immunophenotype, genotype and functional features. Truly malignant cell lines must be discerned from Epstein-Barr virus (EBV)-immortalized normal cells, using various distinguishing parameters. However, the picture is not quite so straightforward, as some types of LL cell lines are indeed EBV+, and some EBV+ normal cell lines carry also genetic aberrations and may mimic malignancy-associated features. Apart from EBV and human T-cell leukemia virus in some lines, the majority of wild-type LL cell lines are virus-negative. The efficiency of cell line establishment is rather low and the deliberate establishment of new LL cell lines remains by and large an unpredictable random process. Difficulties in establishing continuous cell lines may be caused by the inappropriate selection of nutrients and growth factors for these cells. Clearly, a generally suitable microenvironment for hematopoietic cells, either malignant or normal, cannot yet be created in vitro. The characterization and publication of new LL cell

  6. Tuft (caveolated) cells in two human colon carcinoma cell lines.

    PubMed Central

    Barkla, D. H.; Whitehead, R. H.; Foster, H.; Tutton, P. J.

    1988-01-01

    The presence of an unusual cell type in two human colon carcinoma cell lines is reported. The cells show the same morphology as "tuft" (caveolated) cells present in normal gastrointestinal epithelium. Tuft cells were seen in cell line LIM 1863 growing in vitro and in human colon carcinoma cell line LIM 2210 growing as subcutaneous solid tumour xenografts in nude mice. Characteristic morphologic features of tuft cells included a wide base, narrow apex and a tuft of long microvilli projecting from the apical surface. The microvilli are attached by a core of long microfilaments passing deep into the apical cytoplasm. Between the microvilli are parallel arrays of vesicles (caveoli) containing flocculent material. Two different but not mutually exclusive explanations for the presence of tuft cells are proposed. The first explanation is that tuft cells came from the resected tumour and have survived by mitotic division during subsequent passages. The second explanation suggests that tuft cells are the progeny of undifferentiated tumour cells. Descriptions of tuft cells in colon carcinomas are uncommon and possible reasons for this are presented. The morphology of tuft cells is consistent with that of a highly differentiated cell specialised for absorption, and these new models provide an opportunity to further investigate the structure and function of tuft cells. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 PMID:3414781

  7. Effects of Cabazitaxel in Renal Cell Carcinoma Cell Lines.

    PubMed

    Mizutani, Kosuke; Tomoda, Masashi; Ohno, Yuta; Hayashi, Hideki; Fujita, Yasunori; Kawakami, Kyojiro; Kameyama, Koji; Kato, Taku; Sugiyama, Tadashi; Itoh, Yoshinori; Ito, Masafumi; Deguchi, Takashi

    2015-12-01

    Advanced renal cell carcinoma is treated with mammalian target of rapamycin (mTOR) inhibitors or tyrosine kinase inhibitors (TKIs). The effects of these drugs are, however, limited and novel treatment strategies are required. Clear-cell type renal cell carcinoma (ccRCC) is chemo-resistant, in part, due to expression of multidrug resistance proteins such as p-glycoprotein. Cabazitaxel, a tubulin-binding taxane drug used for castration-resistant prostate cancer, has less affinity for p-glycoprotein compared to docetaxel. In the current study, the effects of docetaxel and cabazitaxel on ccRCC cells were investigated. The expression of p-glycoprotein was evaluated in the ccRCC cell lines, Caki-1, KMRC-1 and OS-RC-2 by western blotting. Cells were treated with cabazitaxel or docetaxel, and growth kinetics and tubulin polymerization were determined by the WST-1 assay and cell-based tubulin polymerization assay, respectively. Intracellular drug concentrations were measured by chromatography. AKT activation after treatment was examined by western blotting. All ccRCC cell lines expressed p-glycoprotein. Cabazitaxel inhibited cell growth and induced tubulin polymerization more potently than docetaxel. The intracellular concentration of cabazitaxel was much higher than docetaxel in all cell lines. Both docetaxel and cabazitaxel inhibit AKT phosphorylation at 5 min among three cells. Cabazitaxel inhibits growth of ccRCC cells expressing p-glycoprotein and could thus be possibly used for advanced ccRCC patients in combination with targeted-therapy enhancing their effects. Copyright© 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

  8. Radiation sensitivities of 31 human oesophageal squamous cell carcinoma cell lines

    PubMed Central

    Ban, Sadayuki; Michikawa, Yuichi; Ishikawa, Ken-ichi; Sagara, Masashi; Watanabe, Koji; Shimada, Yutaka; Inazawa, Johji; Imai, Takashi

    2005-01-01

    The purpose of this study was to determine the radiosensitivities of 31 human oesophageal squamous cell carcinoma cell lines with a colony-formation assay. A large variation in radiosensitivity existed among 31 cell lines. Such a large variation may partly explain the poor result of radiotherapy for this cancer. One cell line (KYSE190) demonstrated an unusual radiosensitivity. Ataxia-telangiectasia-mutated (ATM) gene in these cells had five missense mutations, and ATM protein was truncated or degraded. Inability to phosphorylate Chk2 in the irradiated KYSE190 cells suggests that the ATM protein in these cells had lost its function. The dysfunctional ATM protein may be a main cause of unusual radiosensitivity of KYSE190 cells. Because the donor of these cells was not diagnosed with ataxia telangiectasia, mutations in ATM gene might have occurred during the initiation and progression of cancer. Radiosensitive cancer developed in non-hereditary diseased patients must be a good target for radiotherapy. PMID:16045545

  9. Evaluating cell lines as tumour models by comparison of genomic profiles

    PubMed Central

    Domcke, Silvia; Sinha, Rileen; Levine, Douglas A.; Sander, Chris; Schultz, Nikolaus

    2013-01-01

    Cancer cell lines are frequently used as in vitro tumour models. Recent molecular profiles of hundreds of cell lines from The Cancer Cell Line Encyclopedia and thousands of tumour samples from the Cancer Genome Atlas now allow a systematic genomic comparison of cell lines and tumours. Here we analyse a panel of 47 ovarian cancer cell lines and identify those that have the highest genetic similarity to ovarian tumours. Our comparison of copy-number changes, mutations and mRNA expression profiles reveals pronounced differences in molecular profiles between commonly used ovarian cancer cell lines and high-grade serous ovarian cancer tumour samples. We identify several rarely used cell lines that more closely resemble cognate tumour profiles than commonly used cell lines, and we propose these lines as the most suitable models of ovarian cancer. Our results indicate that the gap between cell lines and tumours can be bridged by genomically informed choices of cell line models for all tumour types. PMID:23839242

  10. Assessment of geraniol-incorporated polymers to control Aedes albopictus (Diptera: culicidae)

    PubMed Central

    Chuaycharoensuk, T.; Manguin, S.; Duvallet, G.; Chareonviriyaphap, T.

    2012-01-01

    Effective control of mosquito borne diseases has proven extremely difficult with both vector and pathogen remaining entrenched and expanding in many disease endemic areas. When lacking an effective vaccine, vector control methods targeting both larval habitats and adult mosquito populations remain the primary strategy for reducing risk. Aedes albopictus from Thailand was used as a reference baseline for evaluation of natural insecticides incorporated in polymer disks and pellets and tested both in laboratory and field conditions. In laboratory and field tests, the highest larval mortality was obtained with disks or pellets containing IKHC (Insect Killer Highly Concentrate) from Fulltec AG Company. This product is reputed to contain geraniol as an active ingredient. With pellets, high mortality of Ae. albopictus larvae (92%) was observed in presence of 1 g of pellets per 500 ml of water at day 1st, and the mortality was 100% at day 1st for larvae in presence of 5 or 10 g of pellets. Fulltec AG Company has not accepted to give us the exact composition of their IKHC product. Therefore, we cannot recommend it, but the principle of using monoterpenes like geraniol, incorporated into polymer disks or pellets as natural larvicide needs more attention as it could be considered as a powerful alternative in mosquito vector control. PMID:22910616

  11. Peridomestic Aedes malayensis and Aedes albopictus are capable vectors of arboviruses in cities.

    PubMed

    Mendenhall, Ian H; Manuel, Menchie; Moorthy, Mahesh; Lee, Theodore T M; Low, Dolyce H W; Missé, Dorothée; Gubler, Duane J; Ellis, Brett R; Ooi, Eng Eong; Pompon, Julien

    2017-06-01

    Dengue and chikungunya are global re-emerging mosquito-borne diseases. In Singapore, sustained vector control coupled with household improvements reduced domestic mosquito populations for the past 45 years, particularly the primary vector Aedes aegypti. However, while disease incidence was low for the first 30 years following vector control implementation, outbreaks have re-emerged in the past 15 years. Epidemiological observations point to the importance of peridomestic infection in areas not targeted by control programs. We investigated the role of vectors in peri-domestic areas. We carried out entomological surveys to identify the Aedes species present in vegetated sites in highly populated areas and determine whether mosquitoes were present in open-air areas frequented by people. We compared vector competence of Aedes albopictus and Aedes malayensis with Ae. aegypti after oral infection with sympatric dengue serotype 2 and chikungunya viruses. Mosquito saliva was tested for the presence of infectious virus particles as a surrogate for transmission following oral infection. We identified Aedes albopictus and Aedes malayensis throughout Singapore and quantified their presence in forested and opened grassy areas. Both Ae. albopictus and Ae. malayensis can occupy sylvatic niches and were highly susceptible to both arboviruses. A majority of saliva of infected Ae. malayensis contained infectious particles for both viruses. Our study reveals the prevalence of competent vectors in peri-domestic areas, including Ae. malayensis for which we established the vector status. Epidemics can be driven by infection foci, which are epidemiologically enhanced in the context of low herd immunity, selective pressure on arbovirus transmission and the presence of infectious asymptomatic persons, all these conditions being present in Singapore. Learning from Singapore's vector control success that reduced domestic vector populations, but has not sustainably reduced arboviral incidence

  12. Peridomestic Aedes malayensis and Aedes albopictus are capable vectors of arboviruses in cities

    PubMed Central

    Manuel, Menchie; Low, Dolyce H. W.; Missé, Dorothée; Gubler, Duane J.; Ellis, Brett R.; Ooi, Eng Eong; Pompon, Julien

    2017-01-01

    Background Dengue and chikungunya are global re-emerging mosquito-borne diseases. In Singapore, sustained vector control coupled with household improvements reduced domestic mosquito populations for the past 45 years, particularly the primary vector Aedes aegypti. However, while disease incidence was low for the first 30 years following vector control implementation, outbreaks have re-emerged in the past 15 years. Epidemiological observations point to the importance of peridomestic infection in areas not targeted by control programs. We investigated the role of vectors in peri-domestic areas. Methods We carried out entomological surveys to identify the Aedes species present in vegetated sites in highly populated areas and determine whether mosquitoes were present in open-air areas frequented by people. We compared vector competence of Aedes albopictus and Aedes malayensis with Ae. aegypti after oral infection with sympatric dengue serotype 2 and chikungunya viruses. Mosquito saliva was tested for the presence of infectious virus particles as a surrogate for transmission following oral infection. Results We identified Aedes albopictus and Aedes malayensis throughout Singapore and quantified their presence in forested and opened grassy areas. Both Ae. albopictus and Ae. malayensis can occupy sylvatic niches and were highly susceptible to both arboviruses. A majority of saliva of infected Ae. malayensis contained infectious particles for both viruses. Conclusions Our study reveals the prevalence of competent vectors in peri-domestic areas, including Ae. malayensis for which we established the vector status. Epidemics can be driven by infection foci, which are epidemiologically enhanced in the context of low herd immunity, selective pressure on arbovirus transmission and the presence of infectious asymptomatic persons, all these conditions being present in Singapore. Learning from Singapore’s vector control success that reduced domestic vector populations, but has

  13. Evidence of Zika Virus RNA Fragments in Aedes albopictus (Diptera: Culicidae) Field-Collected Eggs From Camaçari, Bahia, Brazil.

    PubMed

    Smartt, Chelsea T; Stenn, Tanise M S; Chen, Tse-Yu; Teixeira, Maria Gloria; Queiroz, Erivaldo P; Souza Dos Santos, Luciano; Queiroz, Gabriel A N; Ribeiro Souza, Kathleen; Kalabric Silva, Luciano; Shin, Dongyoung; Tabachnick, Walter J

    2017-07-01

    A major mosquito-borne viral disease outbreak caused by Zika virus (ZIKV) occurred in Bahia, Brazil, in 2015, largely due to transmission by the mosquito, Aedes aegypti (L.). Detecting ZIKV in field samples of Ae. aegypti has proven problematic in some locations, suggesting other mosquito species might be contributing to the spread of ZIKV. In this study, several (five) adult Aedes albopictus (Skuse) mosquitoes that emerged from a 2015 field collection of eggs from Camaçari, Bahia, Brazil, were positive for ZIKV RNA; however, attempts to isolate live virus were not successful. Results from this study suggest that field-collected Ae. albopictus eggs may contain ZIKV RNA that require further tests for infectious ZIKV. There is a need to investigate the role of Ae. albopictus in the ZIKV infection process in Brazil and to study the potential presence of vertical and sexual transmission of ZIKV in this species. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  14. Effects of Infectious Virus Dose and Bloodmeal Delivery Method on Susceptibility of Aedes aegypti and Aedes albopictus to Chikungunya Virus

    PubMed Central

    Pesko, Kendra; Westbrook, Catherine J.; Mores, Christopher N.; Lounibos, L. Philip; Reiskind, Michael H.

    2009-01-01

    Chikungunya virus (CHIKV) is an arbovirus (genus Alphavirus, family Togaviridae) that has recently caused disease outbreaks in the Indian Ocean basin and southern Europe. These outbreaks could be associated with a possible shift in primary vector from Aedes aegypti to Ae. albopictus. To evaluate vector competence differences in possible CHIKV vectors, we evaluated the dose-dependant susceptibility of Florida strains of Ae. albopictus and Ae. aegypti for infection with a La Réunion island strain of CHIKV. Pledget and water-jacketed membrane feeding systems were also evaluated. We show that both Aedes spp. were susceptible to the highest CHIKV doses, whereas only Ae. albopictus developed disseminated infections after exposure to the two lowest doses. Infection rates for both mosquito species were significantly affected by the bloodmeal delivery method used. This information is important in assessing risk of an outbreak of imported CHIKV in the United States, in determining differences in vectorial capacity of these two vector species, and in evaluating arbovirus delivery methods in the laboratory. PMID:19351094

  15. First record of Aedes albopictus (Skuse 1894) on São tomé island.

    PubMed

    Reis, Sandra; Cornel, Anthony J; Melo, Martim; Pereira, Hugo; Loiseau, Claire

    2017-07-01

    Biological invasions have critical impacts on native biodiversity and human societies and especially on oceanic islands that are fragile and threatened ecosystems. The invasive tiger mosquito Aedes (Stegomyia) albopictus (Skuse, 1894) native to Southeast Asia has been introduced during the past 30 years almost everywhere in the world, including the Americas, the Pacific, Europe and Africa. It has been reported for the first time in the Gulf of Guinea in 2000, first in Cameroon, then in Bioko Island in 2003 and more recently in Gabon in 2007. Here we report the first record of Ae. albopictus on São Tomé Island. Although we cannot estimate precisely the year of introduction on São Tomé Island, it most likely arrived within the last 10 years. By sequencing the mitochondrial cytochrome c oxidase gene from individual adults, we detected three haplotypes already present in mainland Africa. More studies are needed to explore the dynamics of its expansion and competition with insular native mosquitoes. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Invasiveness of Aedes aegypti and Aedes albopictus and Vectorial Capacity for Chikungunya Virus.

    PubMed

    Lounibos, Leon Philip; Kramer, Laura D

    2016-12-15

    In this review, we highlight biological characteristics of Aedes aegypti and Aedes albopictus, 2 invasive mosquito species and primary vectors of chikungunya virus (CHIKV), that set the tone of these species' invasiveness, vector competence, and vectorial capacity (VC). The invasiveness of both species, as well as their public health threats as vectors, is enhanced by preference for human blood. Vector competence, characterized by the efficiency of an ingested arbovirus to replicate and become infectious in the mosquito, depends largely on vector and virus genetics, and most A. aegypti and A. albopictus populations thus far tested confer vector competence for CHIKV. VC, an entomological analog of the pathogen's basic reproductive rate (R 0 ), is epidemiologically more important than vector competence but less frequently measured, owing to challenges in obtaining valid estimates of parameters such as vector survivorship and host feeding rates. Understanding the complexities of these factors will be pivotal in curbing CHIKV transmission. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  17. Host-feeding pattern of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in heterogeneous landscapes of South Andaman, Andaman and Nicobar Islands, India.

    PubMed

    Sivan, Arun; Shriram, A N; Sunish, I P; Vidhya, P T

    2015-09-01

    Mosquito foraging behavior is a determinant of host-vector contact and has an impact on the risk of arboviral epidemics. Therefore, blood-feeding patterns is a useful tool for assessing the role in pathogen transmission by vector mosquitoes. Competent vectors of dengue and chikungunya viz. Aedes aegypti and Aedes albopictus are widely prevalent in the Andaman and Nicobar archipelago. Considering the vector potential, medical importance of both these mosquito species and lack of information on host-feeding patterns, blood meal analysis of both these vector mosquitoes was undertaken. Biogents Sentinel traps were used for sampling blooded mosquitoes, for identifying the source of blood meal by agar gel-precipitin test. We identified vertebrate source of 147 and 104 blood meals in Ae. aegypti and Ae. albopictus from heterogeneous landscapes in South Andaman district. Results revealed that Ae. aegypti (88 %) and Ae. albopictus (49 %) fed on human and a small proportion on mammals and fowls, indicative of predominance of anthropophilism. Ae. aegypti predominantly fed on human blood (94.2 %-densely built urban, 89.8 %-low vegetation coverage, and 78.3 %-medium vegetation coverage). Anthropophilism in Ae. albopictus was maximal in densely built urban (90.5 %) and progressively decreased from low vegetation-vegetation/forested continuum (66.7, 36.4, and 8.7 %), indicating plasticity in feeding across these landscapes. Epidemiological significance of the findings is discussed.

  18. Tracing the Tiger: Population Genetics Provides Valuable Insights into the Aedes (Stegomyia) albopictus Invasion of the Australasian Region

    PubMed Central

    Beebe, Nigel W.; Ambrose, Luke; Hill, Lydia A.; Davis, Joseph B.; Hapgood, George; Cooper, Robert D.; Russell, Richard C.; Ritchie, Scott A.; Reimer, Lisa J.; Lobo, Neil F.; Syafruddin, Din; van den Hurk, Andrew F.

    2013-01-01

    Background The range of the Asian tiger mosquito Aedes albopictus is expanding globally, raising the threat of emerging and re-emerging arbovirus transmission risks including dengue and chikungunya. Its detection in Papua New Guinea's (PNG) southern Fly River coastal region in 1988 and 1992 placed it 150 km from mainland Australia. However, it was not until 12 years later that it appeared on the Torres Strait Islands. We hypothesized that the extant PNG population expanded into the Torres Straits as an indirect effect of drought-proofing the southern Fly River coastal villages in response to El Nino-driven climate variability in the region (via the rollout of rainwater tanks and water storage containers). Methodology/Principal Findings Examination of the mosquito's mitochondrial DNA cytochrome oxidase I (COI) sequences and 13 novel nuclear microsatellites revealed evidence of substantial intermixing between PNG's southern Fly region and Torres Strait Island populations essentially compromising any island eradication attempts due to potential of reintroduction. However, two genetically distinct populations were identified in this region comprising the historically extant PNG populations and the exotic introduced population. Both COI sequence data and microsatellites showed the introduced population to have genetic affinities to populations from Timor Leste and Jakarta in the Indonesian region. Conclusions/Significance The Ae. albopictus invasion into the Australian region was not a range expansion out of PNG as suspected, but founded by other, genetically distinct population(s), with strong genetic affinities to populations sampled from the Indonesian region. We now suspect that the introduction of Ae. albopictus into the Australian region was driven by widespread illegal fishing activity originating from the Indonesian region during this period. Human sea traffic is apparently shuttling this mosquito between islands in the Torres Strait and the southern PNG

  19. Multi-country Survey Revealed Prevalent and Novel F1534S Mutation in Voltage-Gated Sodium Channel (VGSC) Gene in Aedes albopictus.

    PubMed

    Xu, Jiabao; Bonizzoni, Mariangela; Zhong, Daibin; Zhou, Guofa; Cai, Songwu; Li, Yiji; Wang, Xiaoming; Lo, Eugenia; Lee, Rebecca; Sheen, Roger; Duan, Jinhua; Yan, Guiyun; Chen, Xiao-Guang

    2016-05-01

    Aedes albopictus is an important dengue vector because of its aggressive biting behavior and rapid spread out of its native home range in Southeast Asia. Pyrethroids are widely used for adult mosquito control, and resistance to pyrethroids should be carefully monitored because vector control is the only effective method currently available to prevent dengue transmission. The voltage-gated sodium channel gene is the target site of pyrethroids, and mutations in this gene cause knockdown resistance (kdr). Previous studies reported various mutations in the voltage-gated sodium channel (VGSC) gene, but the spatial distribution of kdr mutations in Ae. albopictus has not been systematically examined, and the association between kdr mutation and phenotypic resistance has not been established. A total of 597 Ae. albopictus individuals from 12 populations across Asia, Africa, America and Europe were examined for mutations in the voltage-gated sodium channel gene. Three domains for a total of 1,107 bp were sequenced for every individual. Two populations from southern China were examined for pyrethroid resistance using the World Health Organization standard tube bioassay, and the association between kdr mutations and phenotypic resistance was tested. A total of 29 synonymous mutations were found across domain II, III and IV of the VGSC gene. Non-synonymous mutations in two codons of the VGSC gene were detected in 5 populations from 4 countries. A novel mutation at 1532 codon (I1532T) was found in Rome, Italy with a frequency of 19.7%. The second novel mutation at codon 1534 (F1534S) was detected in southern China and Florida, USA with a frequency ranging from 9.5-22.6%. The WHO insecticide susceptibility bioassay found 90.1% and 96.1% mortality in the two populations from southern China, suggesting resistance and probable resistance. Positive association between kdr mutations with deltamethrin resistance was established in these two populations. Two novel kdr mutations, I1532T

  20. Establishment and characterization of three immortal bovine muscular epithelial cell lines.

    PubMed

    Jin, Xun; Lee, Joong-Seob; Kwak, Sungwook; Lee, Soo-Yeon; Jung, Ji-Eun; Kim, Tae-Kyung; Xu, Chenxiong; Hong, Zhongshan; Li, Zhehu; Kim, Sun-Myung; Pian, Xumin; Lee, Dong-Hee; Yoon, Jong-Taek; You, Seungkwon; Choi, Yun-Jaie; Kim, Huunggee

    2006-02-28

    We have established three immortal bovine muscular epithelial (BME) cell lines, one spontaneously immortalized (BMES), the second SV40LT-mediated (BMEV) and the third hTERT-mediated (BMET). The morphology of the three immortal cell lines was similar to that of early passage primary BME cells. Each of the immortal cell lines made cytokeratin, a typical epithelial marker. BMET grew faster than the other immortal lines and the BME cells, in 10% FBS-DMEM medium, whereas neither the primary cells nor the three immortal cell lines grew in 0.5% FBS-DMEM. The primary BME cells and the immortal cell lines, with the exception of BMES, made increasing amounts of p53 protein when treated with doxorubicin, a DNA damaging agent. On the other hand, almost half of the cells in populations of the three immortal cell lines may lack p16(INK4a) regulatory function, compared to primary BME cells that were growth arrested by enforced expression of p16(INK4a). In soft-agar assays, the primary cells and immortal cell lines proved to be less transformed in phenotype than HeLa cells. The three immortal epithelial-type cell lines reported here are the first cell lines established from muscle tissue of bovine or other species.

  1. Terra and Aqua satellites track tiger mosquito invasion: modelling the potential distribution of Aedes albopictus in north-eastern Italy

    PubMed Central

    2011-01-01

    Background The continuing spread of the Asian tiger mosquito Aedes albopictus in Europe is of increasing public health concern due to the potential risk of new outbreaks of exotic vector-borne diseases that this species can transmit as competent vector. We predicted the most favorable areas for a short term invasion of Ae. albopictus in north-eastern Italy using reconstructed daily satellite data time series (MODIS Land Surface Temperature maps, LST). We reconstructed more than 11,000 daily MODIS LST maps for the period 2001-09 (i.e. performed spatial and temporal gap-filling) in an Open Source GIS framework. We aggregated these LST maps over time and identified the potential distribution areas of Ae. albopictus by adapting published temperature threshold values using three variables as predictors (0°C for mean January temperatures, 11°C for annual mean temperatures and 1350 growing degree days filtered for areas with autumnal mean temperatures > 11°C). The resulting maps were integrated into the final potential distribution map and this was compared with the known current distribution of Ae. albopictus in north-eastern Italy. Results LST maps show the microclimatic characteristics peculiar to complex terrains, which would not be visible in maps commonly derived from interpolated meteorological station data. The patterns of the three indicator variables partially differ from each other, while winter temperature is the determining limiting factor for the distribution of Ae. albopictus. All three variables show a similar spatial pattern with some local differences, in particular in the northern part of the study area (upper Adige valley). Conclusions Reconstructed daily land surface temperature data from satellites can be used to predict areas of short term invasion of the tiger mosquito with sufficient accuracy (200 m pixel resolution size). Furthermore, they may be applied to other species of arthropod of medical interest for which temperature is a relevant

  2. Derivation and characterization of a pig embryonic stem cell-derived exocrine pancreatic cell line

    USDA-ARS?s Scientific Manuscript database

    The establishment and initial characterization of a pig embryonic stem cell-derived pancreatic cell line, PICM-31, and a colony-cloned derivative cell line, PICM-31A, is described. The cell lines were propagated for several months at split ratios of 1:3 or 1:5 at each passage on STO feeder cells af...

  3. Spatiotemporal oviposition and habitat preferences of Ochlerotatus triseriatus and Aedes albopictus in an emerging focus of La Crosse virus.

    PubMed

    Barker, Christopher M; Brewster, Carlyle C; Paulson, Sally L

    2003-12-01

    The number of cases of encephalitis caused by La Crosse virus recently has increased in southwestern Virginia counties. This article presents results of a study conducted from May to September 2000 in Wise County, VA, that examined the area-wide oviposition activity and habitat preferences of Ochlerotatus triseriatus and Aedes albopictus, potential vectors of La Crosse virus in the region. Data from 490 ovitrap collections made throughout the county showed that mean oviposition activity throughout the study was higher for Oc. triseriatus (20.4 eggs/trap-day) than for Ae. albopictus (3.7 eggs/trap-day). The 2 species also had distinct habitat preferences for oviposition, with Oc. triseriatus favoring forested habitats and Ae. albopictus favoring urban/residential habitats. A landcover map of 6 habitat types derived from Landsat satellite imagery of the county showed that 63% of the county was forested and 18% was urban/residential. A Bayesian decision-rule model that incorporated the ovitrap data and landcover map was moderately successful at predicting the occurrence of high oviposition activity and abundance of the 2 species. The predictions reflected seasonal and spatial fluctuations in oviposition activity, with accuracies between 55 and 79% for Oc. triseriatus and 70 and 94% for Ae. albopictus. Kappa (K), a measure of the predictive power of the model, varied from poor (K < 0.4) to good (0.4 < K < 0.75) for both species, and was highest during periods when actual egg abundance was high. This suggests that the predictions were most accurate during periods when the risk for La Crosse virus transmission is greatest. Limitations and suggestions for improving the model are discussed.

  4. Spinosad as an effective larvicide for control of Aedes albopictus and Aedes aegypti, vectors of dengue in southern Mexico.

    PubMed

    Marina, Carlos F; Bond, J Guillermo; Casas, Mauricio; Muñoz, José; Orozco, Arnoldo; Valle, Javier; Williams, Trevor

    2011-01-01

    Field trials were conducted during the wet and dry seasons in periurban and semi-rural cemeteries in southern Mexico to determine the efficacy of a suspension concentrate formulation of spinosad (Tracer 480SC) on the inhibition of development of Aedes albopictus L. and Ae. aegypti Skuse. For this, oviposition traps were treated with spinosad (1 or 5 mg L(-1)), Bacillus thuringiensis israelensis (Bti, VectoBac 12AS), a sustained release formulation of temephos and a water control. Ae. albopictus was subordinate to Ae. aegypti during the dry season, but became dominant or codominant during the wet season at both sites. The two species could not be differentiated in field counts on oviposition traps. Mean numbers of larvae + pupae of Aedes spp. in Bti-treated containers were similar to the control at both sites during both seasons. The duration of complete absence of aquatic stages varied from 5 to 13 weeks for the spinosad treatments and from 6 to 9 weeks for the temephos treatment, depending on site, season and product concentration. Predatory Toxorhynchites theobaldi Dyar and Knab suffered low mortality in control and Bti treatments, but high mortality in spinosad and temephos treatments. Egg counts and percentage of egg hatch of Aedes spp. increased significantly between the dry and wet seasons, but significant treatment differences were not detected. Temephos granules and a suspension concentrate formulation of spinosad were both highly effective larvicides against Ae. aegypti and Ae. albopictus. These compounds merit detailed evaluation for inclusion in integrated control programs targeted at Ae. aegypti and Ae. albopictus in regions where they represent important vectors of human diseases. Copyright © 2010 Society of Chemical Industry.

  5. Genetic evidence for a worldwide chaotic dispersion pattern of the arbovirus vector, Aedes albopictus.

    PubMed

    Manni, Mosè; Guglielmino, Carmela R; Scolari, Francesca; Vega-Rúa, Anubis; Failloux, Anna-Bella; Somboon, Pradya; Lisa, Antonella; Savini, Grazia; Bonizzoni, Mariangela; Gomulski, Ludvik M; Malacrida, Anna R; Gasperi, Giuliano

    2017-01-01

    Invasive species represent a global concern for their rapid spread and the possibility of infectious disease transmission. This is the case of the global invader Aedes albopictus, the Asian tiger mosquito. This species is a vector of medically important arboviruses, notably chikungunya (CHIKV), dengue (DENV) and Zika (ZIKV). The reconstruction of the complex colonization pattern of this mosquito has great potential for mitigating its spread and, consequently, disease risks. Classical population genetics analyses and Approximate Bayesian Computation (ABC) approaches were combined to disentangle the demographic history of Aedes albopictus populations from representative countries in the Southeast Asian native range and in the recent and more recently colonized areas. In Southeast Asia, the low differentiation and the high co-ancestry values identified among China, Thailand and Japan indicate that, in the native range, these populations maintain high genetic connectivity, revealing their ancestral common origin. China appears to be the oldest population. Outside Southeast Asia, the invasion process in La Réunion, America and the Mediterranean Basin is primarily supported by a chaotic propagule distribution, which cooperates in maintaining a relatively high genetic diversity within the adventive populations. From our data, it appears that independent and also trans-continental introductions of Ae. albopictus may have facilitated the rapid establishment of adventive populations through admixture of unrelated genomes. As a consequence, a great amount of intra-population variability has been detected, and it is likely that this variability may extend to the genetic mechanisms controlling vector competence. Thus, in the context of the invasion process of this mosquito, it is possible that both population ancestry and admixture contribute to create the conditions for the efficient transmission of arboviruses and for outbreak establishment.

  6. Polymer encapsulated dopaminergic cell lines as "alternative neural grafts".

    PubMed

    Jaeger, C B; Greene, L A; Tresco, P A; Winn, S R; Aebischer, P

    1990-01-01

    Our preliminary findings (Jaeger et al., 1988; Aebischer et al., 1989; Tresco et al., 1989) and the studies in progress show that encapsulated dopaminergic cell lines survive enclosure within a semi-permeable membrane. The encapsulated cells remained viable for extended time periods when maintained in vitro. Moreover, encapsulated PC12 and T28 cells have the potential to survive following their implantation into the forebrain of rats. Cell lines are essentially "immortal" because they continue to divide indefinitely. This property allows perpetual "self-renewal" of a given cell population. However, the capacity of continuous uncontrolled cell division may also lead to tumor formation. This in fact is the case for unencapsulated PC12 cell implants placed into the brain of young Sprague Dawley rats (Jaeger, 1985). Cell line encapsulation has the potential to prevent tumor growth (Jaeger et al., 1988). Survival for 6 months in vitro suggests that encapsulation does not preclude long-term maintenance of an homogeneous cell line like PC12 cells. The presence of mitotic figures in the capsules further supports the likelihood of propagation and self renewal of the encapsulated population. Another significant property of cell lines is that they consist of a single, genetically homogeneous cell type. They do not require specific synaptic interactions for their survival. In the case of PC12 and T28 lines, the cells synthesize and release neurotransmitters. Our data show that PC12 and T28 cells continue to release dopamine spontaneously and to express specific transmitters and enzymes following encapsulation. Thus, cell lines such as these may constitute relatively simple "neural implants" exerting their function via humoral release.(ABSTRACT TRUNCATED AT 250 WORDS)

  7. Discovery of HeLa Cell Contamination in HES Cells: Call for Cell Line Authentication in Reproductive Biology Research.

    PubMed

    Kniss, Douglas A; Summerfield, Taryn L

    2014-08-01

    Continuous cell lines are used frequently in reproductive biology research to study problems in early pregnancy events and parturition. It has been recognized for 50 years that many mammalian cell lines contain inter- or intraspecies contaminations with other cells. However, most investigators do not routinely test their culture systems for cross-contamination. The most frequent contributor to cross-contamination of cell lines is the HeLa cell isolated from an aggressive cervical adenocarcinoma. We report on the discovery of HeLa cell contamination of the human endometrial epithelial cell line HES isolated in our laboratory. Short tandem repeat analysis of 9 unique genetic loci demonstrated molecular identity between HES and HeLa cells. In addition, we verified that WISH cells, isolated originally from human amnion epithelium, were also contaminated with HeLa cells. Inasmuch as our laboratory did not culture HeLa cells at the time of HES cell derivations, the source of contamination was the WISH cell line. These data highlight the need for continued diligence in authenticating cell lines used in reproductive biology research. © The Author(s) 2014.

  8. THP-1 cell line: an in vitro cell model for immune modulation approach.

    PubMed

    Chanput, Wasaporn; Mes, Jurriaan J; Wichers, Harry J

    2014-11-01

    THP-1 is a human leukemia monocytic cell line, which has been extensively used to study monocyte/macrophage functions, mechanisms, signaling pathways, and nutrient and drug transport. This cell line has become a common model to estimate modulation of monocyte and macrophage activities. This review attempts to summarize and discuss recent publications related to the THP-1 cell model. An overview on the biological similarities and dissimilarities between the THP-1 cell line and human peripheral blood mononuclear cell (PBMC) derived-monocytes and macrophages, as well as the advantages and disadvantages of the use of THP-1 cell line, is included. The review summarizes different published co-cultivation studies of THP-1 cells with other cell types, for instance, intestinal cells, adipocytes, T-lymphocytes, platelets, and vascular smooth muscle cells, which can be an option to study cell-cell interaction in vitro and can be an approach to better mimic in vivo conditions. Macrophage polarization is a relatively new topic which gains interest for which the THP-1 cell line also may be relevant. Besides that an overview of newly released commercial THP-1 engineered-reporter cells and THP-1 inflammasome test-cells is also given. Evaluation of recent papers leads to the conclusion that the THP-1 cell line has unique characteristics as a model to investigate/estimate immune-modulating effects of compounds in both activated and resting conditions of the cells. Although the THP-1 response can hint to potential responses that might occur ex vivo or in vivo, these should be, however, validated by in vivo studies to draw more definite conclusions. Copyright © 2013. Published by Elsevier B.V.

  9. Precipitation and Temperature Effects on Populations of Aedes albopictus (Diptera: Culicidae): Implications for Range Expansion

    PubMed Central

    ALTO, BARRY W.; JULIANO, STEVEN A.

    2008-01-01

    We investigated how temperature and precipitation regime encountered over the life cycle of Aedes albopictus (Skuse) affects populations. Caged populations of A. albopictus were maintained at 22, 26, and 30°C. Cages were equipped with containers that served as sites for oviposition and larval development. All cages were assigned to one of three simulated precipitation regimes: (1) low fluctuation regime - water within the containers was allowed to evaporate to 90% of its maximum before being refilled, (2) high fluctuation regime - water was allowed to evaporate to 25% of its maximum before being refilled, and (3) drying regime - water was allowed to evaporate to complete container dryness before being refilled. Greater temperature and the absence of drying resulted in greater production of adults. Greater temperature in combination with drying were detrimental to adult production. These precipitation effects on adult production were absent at 22°C. Greater temperatures and drying treatments yielded higher and lower eclosion rates, respectively and, both yielded greater mortality. Development time and size of adults decreased with increased temperatures, and drying produced larger adults. Greater temperatures resulted in greater egg mortality. These results suggest that populations occurring in warmer regions are likely to produce more adults as long as containers do not dry completely. Populations in cooler regions are likely to produce fewer adults with the variability of precipitation contributing less to variation in adult production. Predicted climate change in North America is likely to extend the northern distribution of A. albopictus and to limit further its establishment in arid regions. PMID:11580037

  10. Presence and Potential Distribution of Aedes albopictus and Aedes japonicus japonicus (Diptera: Culicidae) in Slovenia.

    PubMed

    Kalan, Katja; Ivovic, Vladimir; Glasnovic, Peter; Buzan, Elena

    2017-11-07

    In Slovenia, two invasive mosquito species are present, Aedes albopictus (Skuse, 1895) (Diptera: Culicidae) and Aedes japonicus (Theobald, 1901) (Diptera: Culicidae). In this study, we examined their actual distribution and suitable habitats for new colonizations. Data from survey of species presence in 2013 and 2015, bioclimatic variables and altitude were used for the construction of predictive maps. We produced various models in Maxent software and tested two bioclimatic variable sets, WorldClim and CHELSA. For the variable selection of A. albopictus modeling we used statistical and expert knowledge-based approach, whereas for A. j. japonicus we used only a statistically based approach. The best performing models for both species were chosen according to AIC score-based evaluation. In 2 yr of sampling, A. albopictus was largely confined to the western half of Slovenia, whereas A. j. japonicus spread significantly and can be considered as an established species in a large part of the country. Comparison of models with WorldClim and CHELSA variables for both species showed models with CHELSA variables as a better tool for prediction. Finally, we validated the models performance in predicting distribution of species according to collected field data. Our study confirms that both species are co-occurring and are sympatric in a large part of the country area. The tested models could be used for future prevention of invasive mosquitoes spreading in other countries with similar bioclimatic conditions. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. A Linkage Map of the Asian Tiger Mosquito (Aedes albopictus) Based on cDNA Markers

    PubMed Central

    Sutherland, Ian W.; Mori, Akio; Montgomery, John; Fleming, Karen L.; Anderson, Jennifer M.; Valenzuela, Jesus G.; Severson, David W.

    2011-01-01

    The Asian tiger mosquito, Aedes (Stegomyia) albopictus (Skuse), is an important vector of a number of arboviruses, and populations exhibit extreme variation in adaptive traits such as egg diapause, cold hardiness, and autogeny (ability to mature a batch of eggs without blood feeding). The genetic basis of some of these traits has been established, but lack of a high-resolution linkage map has prevented in-depth genetic analyses of the genes underlying these complex traits. We report here on the breeding of 4 F1 intercross mapping families and the use of these to locate 35 cDNA markers to the A. albopictus linkage map. The present study increases the number of markers on the A. albopictus cDNA linkage map from 38 to 73 and the density of markers from 1 marker/5.7 cM to 1 marker/2.9 cM and adds 9, 16, and 10 markers to the 3 linkage groups, respectively. The overall lengths of the 3 linkage groups are 64.5, 76.5, and 71.6 cM, respectively, for a combined length of 212.6 cM. Despite conservation in the order of most genes among the 4 families and a previous mapping family, we found substantial heterogeneity in the amount of recombination among markers. This was most marked in linkage group I, which varied between 16.7 and 69.3 cM. A map integrating the results from these 4 families with an earlier cDNA linkage map is presented. PMID:21148282

  12. Chemical composition and larvicidal activity of essential oil from Ocimum basilicum (L.) against Culex tritaeniorhynchus, Aedes albopictus and Anopheles subpictus (Diptera: Culicidae).

    PubMed

    Govindarajan, M; Sivakumar, R; Rajeswary, M; Yogalakshmi, K

    2013-05-01

    The toxicity of mosquito larvicidal activity of leaf essential oil and their major chemical constituents from Ocimum basilicum were evaluated against Culex tritaeniorhynchus, Aedes albopictus and Anopheles subpictus. The chemical composition of the leaf essential oil was analyzed using gas chromatography-mass spectroscopy. GC-MS revealed that the essential oil of O. basilicum contained 20 compounds. The major chemical components identified were linalool (52.42%), methyl eugenol (18.74%) and 1, 8-cineol (5.61%). The essential oil had a significant toxic effect against late third-stage larvae of Cx. tritaeniorhynchus, Ae. albopictus and An. subpictus with an LC(50) values of 14.01, 11.97 and 9.75 ppm and an LC(90) values of 23.44, 21.17 and 18.56 ppm, respectively. The results could be useful in search for newer, safer, and more effective natural larvicidal agents against Cx. tritaeniorhynchus, Ae. albopictus and An. subpictus. Copyright © 2013 Elsevier Inc. All rights reserved.

  13. Human Rhabdomyosarcoma Cell Lines for Rhabdomyosarcoma Research: Utility and Pitfalls

    PubMed Central

    Hinson, Ashley R. P.; Jones, Rosanne; Crose, Lisa E. S.; Belyea, Brian C.; Barr, Frederic G.; Linardic, Corinne M.

    2013-01-01

    Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of childhood and adolescence. Despite intergroup clinical trials conducted in Europe and North America, outcomes for high risk patients with this disease have not significantly improved in the last several decades, and survival of metastatic or relapsed disease remains extremely poor. Accrual into new clinical trials is slow and difficult, so in vitro cell-line research and in vivo xenograft models present an attractive alternative for preclinical research for this cancer type. Currently, 30 commonly used human RMS cell lines exist, with differing origins, karyotypes, histologies, and methods of validation. Selecting an appropriate cell line for RMS research has important implications for outcomes. There are also potential pitfalls in using certain cell lines including contamination with murine stromal cells, cross-contamination between cell lines, discordance between the cell line and its associated original tumor, imposter cell lines, and nomenclature errors that result in the circulation of two or more presumed unique cell lines that are actually from the same origin. These pitfalls can be avoided by testing for species-specific isoenzymes, microarray analysis, assays for subtype-specific fusion products, and short tandem repeat analysis. PMID:23882450

  14. Generation of genome-modified Drosophila cell lines using SwAP.

    PubMed

    Franz, Alexandra; Brunner, Erich; Basler, Konrad

    2017-10-02

    The ease of generating genetically modified animals and cell lines has been markedly increased by the recent development of the versatile CRISPR/Cas9 tool. However, while the isolation of isogenic cell populations is usually straightforward for mammalian cell lines, the generation of clonal Drosophila cell lines has remained a longstanding challenge, hampered by the difficulty of getting Drosophila cells to grow at low densities. Here, we describe a highly efficient workflow to generate clonal Cas9-engineered Drosophila cell lines using a combination of cell pools, limiting dilution in conditioned medium and PCR with allele-specific primers, enabling the efficient selection of a clonal cell line with a suitable mutation profile. We validate the protocol by documenting the isolation, selection and verification of eight independently Cas9-edited armadillo mutant Drosophila cell lines. Our method provides a powerful and simple workflow that improves the utility of Drosophila cells for genetic studies with CRISPR/Cas9.

  15. Forecasting the spatial and seasonal dynamic of Aedes albopictus oviposition activity in Albania and Balkan countries.

    PubMed

    Tisseuil, Clément; Velo, Enkelejda; Bino, Silvia; Kadriaj, Perparim; Mersini, Kujtim; Shukullari, Ada; Simaku, Artan; Rogozi, Elton; Caputo, Beniamino; Ducheyne, Els; Della Torre, Alessandra; Reiter, Paul; Gilbert, Marius

    2018-02-01

    The increasing spread of the Asian tiger mosquito, Aedes albopictus, in Europe and US raises public health concern due to the species competence to transmit several exotic human arboviruses, among which dengue, chikungunya and Zika, and urges the development of suitable modeling approach to forecast the spatial and temporal distribution of the mosquito. Here we developed a dynamical species distribution modeling approach forecasting Ae. albopictus eggs abundance at high spatial (0.01 degree WGS84) and temporal (weekly) resolution over 10 Balkan countries, using temperature times series of Modis data products and altitude as input predictors. The model was satisfactorily calibrated and validated over Albania based observed eggs abundance data weekly monitored during three years. For a given week of the year, eggs abundance was mainly predicted by the number of eggs and the mean temperature recorded in the preceding weeks. That is, results are in agreement with the biological cycle of the mosquito, reflecting the effect temperature on eggs spawning, maturation and hatching. The model, seeded by initial egg values derived from a second model, was then used to forecast the spatial and temporal distribution of eggs abundance over the selected Balkan countries, weekly in 2011, 2012 and 2013. The present study is a baseline to develop an easy-handling forecasting model able to provide information useful for promoting active surveillance and possibly prevention of Ae. albopictus colonization in presently non-infested areas in the Balkans as well as in other temperate regions.

  16. [Intergration and epression of porcine endogenous retrovinus in the immortal cell line of Banna Minipig Inberd Line-Mesenhymal Stem Cells].

    PubMed

    Yu, Ping; Liu, Jin; Zhang, Li; Li, Shrng-Fu; Bu, Hong; Li, You-Ping; Cheng, Jing-Qui; Lu, Yan-Rong; Long, Dan

    2005-11-01

    To detect the integration and expression of porcine endogenous retrovirus (PERV) in the immortal cell line of Banna Minipig Inbred Line-Mesenchymal Stem Cells (BMI-MSCs). DNA and total RNA of the immortal cell line of BMI-MSCs were extracted and PCR, RT-PCR were performed to detect PERV-gag, pol and env gene, and the type of PERV was also detected. PERV-gag, pol and env gene were all detected in the primary culture and immortal cell line (passage 150 and passage 180) of BMI-MSCs, and the type of PERV was PERV-A, B. Functional expression of PERV-gag and pol mRNA was also detected. In this laboratory, PERV was not lost during the proceeding of pig inbred and since has been in long-term culture of pig cells in vitro. PERV has integrated into the genome of its natural host, and virus mRNA can effectively express. So it is very essential to evaluate the possibility of xenozoonoses in pig-to-human xenotransplantation.

  17. Standards for Cell Line Authentication and Beyond

    PubMed Central

    Cole, Kenneth D.; Plant, Anne L.

    2016-01-01

    Different genomic technologies have been applied to cell line authentication, but only one method (short tandem repeat [STR] profiling) has been the subject of a comprehensive and definitive standard (ASN-0002). Here we discuss the power of this document and why standards such as this are so critical for establishing the consensus technical criteria and practices that can enable progress in the fields of research that use cell lines. We also examine other methods that could be used for authentication and discuss how a combination of methods could be used in a holistic fashion to assess various critical aspects of the quality of cell lines. PMID:27300367

  18. First report of field evolved resistance to agrochemicals in dengue mosquito, Aedes albopictus (Diptera: Culicidae), from Pakistan.

    PubMed

    Khan, Hafiz Azhar Ali; Akram, Waseem; Shehzad, Khurram; Shaalan, Essam A

    2011-07-22

    Agrochemicals have been widely used in Pakistan for several years. This exposes mosquito populations, particularly those present around agricultural settings, to an intense selection pressure for insecticide resistance. The aim of the present study was to investigate the toxicity of representative agrochemicals against various populations of Aedes albopictus (Skuse) collected from three different regions from 2008-2010. For organophosphates and pyrethroids, the resistance ratios compared with susceptible Lab-PK were in the range of 157-266 fold for chlorpyrifos, 24-52 fold for profenofos, 41-71 fold for triazofos, and 15-26 fold for cypermethrin, 15-53 fold for deltamethrin and 21-58 fold for lambdacyhalothrin. The resistance ratios for carbamates and new insecticides were in the range of 13-22 fold for methomyl, 24-30 fold for thiodicarb, and 41-101 fold for indoxacarb, 14-27 fold for emamectin benzoate and 23-50 fold for spinosad. Pair wise comparisons of the log LC50s of insecticides revealed correlation among several insecticides, suggesting a possible cross resistance mechanism. Moreover, resistance remained stable across 3 years, suggesting field selection for general fitness had also taken place for various populations of Ae. albopictus. Moderate to high level of resistance to agrochemicals in Pakistani field populations of Ae. albopictus is reported here first time. The geographic extent of resistance is unknown but, if widespread, may lead to problems in future vector control.

  19. First evidence of resistance to pyrethroid insecticides in Italian Aedes albopictus populations 26 years after invasion.

    PubMed

    Pichler, Verena; Bellini, Romeo; Veronesi, Rodolfo; Arnoldi, Daniele; Rizzoli, Annapaola; Lia, Riccardo Paolo; Otranto, Domenico; Montarsi, Fabrizio; Carlin, Sara; Ballardini, Marco; Antognini, Elisa; Salvemini, Marco; Brianti, Emanuele; Gaglio, Gabriella; Manica, Mattia; Cobre, Pietro; Serini, Paola; Velo, Enkelejda; Vontas, John; Kioulos, Ilias; Pinto, Joao; Della Torre, Alessandra; Caputo, Beniamino

    2018-06-01

    Aedes albopictus has spread during the last few decades all over the world. This has increased significantly the risk of exotic arbovirus transmission (e.g. chikungunya, dengue, and Zika) also in temperate areas, as demonstrated by the Chikungunya 2007 and 2017 outbreaks in northeastern and central Italy. Insecticides are an important tool for limiting the circulation of these mosquito-borne viruses. The aim of the present study was to address the gap in current knowledge of pyrethroid insecticide resistance of European Ae. albopictus populations, focusing on populations from Italy, Albania and Greece. Bioassays for resistance to permethrin (0.75%), α-cypermethrin (0.05%) or deltamethrin (0.05%) were performed according to World Health Organization (WHO) protocols and showed reduced susceptibility (<90% mortality) of some Italian populations to permethrin and α-cypermethrin, but not to deltamethrin. This study reports the first evidence of resistance to pyrethroids in adult Italian Ae. albopictus populations. Results refer to the season preceding the Chikungunya 2017 outbreak in central Italy and highlight the need to increase efforts to monitor the spread of insecticide resistance and the need to develop strategies to limit the spread of insecticide resistance, particularly in areas where extensive treatments have been carried out to contain disease outbreaks. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  20. Spatial and Temporal Variation in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) Numbers in the Yogyakarta Area of Java, Indonesia, With Implications for Wolbachia Releases.

    PubMed

    Tantowijoyo, W; Arguni, E; Johnson, P; Budiwati, N; Nurhayati, P I; Fitriana, I; Wardana, S; Ardiansyah, H; Turley, A P; Ryan, P; O'Neill, S L; Hoffmann, A A

    2016-01-01

    of mosquito vector populations, particularly through Wolbachia endosymbionts. The success of these strategies depends on understanding the dynamics of vector populations. In preparation for Wolbachia releases around Yogyakarta, we have studied Aedes populations in five hamlets. Adult monitoring with BioGent- Sentinel (BG-S) traps indicated that hamlet populations had different dynamics across the year; while there was an increase in Aedes aegypti (L.) and Aedes albopictus (Skuse) numbers in the wet season, species abundance remained relatively stable in some hamlets but changed markedly (>2 fold) in others. Local rainfall a month prior to monitoring partly predicted numbers of Ae. aegypti but not Ae. albopictus. Site differences in population size indicated by BG-S traps were also evident in ovitrap data. Egg or larval collections with ovitraps repeated at the same location suggested spatial autocorrelation (<250 m) in the areas of the hamlets where Ae. aegypti numbers were high. Overall, there was a weak negative association (r<0.43) between Ae. aegypti and Ae. albopictus numbers in ovitraps when averaged across collections. Ae. albopictus numbers in ovitraps and BG-S traps were positively correlated with vegetation around areas where traps were placed, while Ae. aegypti were negatively correlated with this feature. These data inform intervention strategies by defining periods when mosquito densities are high, highlighting the importance of local site characteristics on populations, and suggesting relatively weak interactions between Ae. aegypti and Ae. albopictus. They also indicate local areas within hamlets where consistently high mosquito densities may influence Wolbachia invasions and other interventions.

  1. Aedes albopictus (Diptera: Culicidae) Oviposition Response to Organic Infusions from Common Flora of Suburban Florida

    USDA-ARS?s Scientific Manuscript database

    We evaluated the oviposition response of Aedes albopictus to six organic infusions. Laboratory and field placed ovitraps baited with water oak (Quercus nigra L.), longleaf pine (Pinus palustris P. Mill) and St. Augustine grass (Stenotaphrum secundatum (Walt.) Kuntze), as well as two-species mixture...

  2. Peptidomic analysis of human cell lines

    PubMed Central

    Gelman, Julia S.; Sironi, Juan; Castro, Leandro M.; Ferro, Emer S.; Fricker, Lloyd D.

    2011-01-01

    Peptides have been proposed to function in intracellular signaling within the cytosol. Although cytosolic peptides are considered to be highly unstable, a large number of peptides have been detected in mouse brain and other biological samples. In the present study, we evaluated the peptidome of three diverse cell lines: SH-SY5Y, MCF7, and HEK293 cells. A comparison of the peptidomes revealed considerable overlap in the identity of the peptides found in each cell line. The majority of the observed peptides are not derived from the most abundant or least stable proteins in the cell, and approximately half of the cellular peptides correspond to the N- or C- termini of the precursor proteins. Cleavage site analysis revealed a preference for hydrophobic residues in the P1 position. Quantitative peptidomic analysis indicated that the levels of most cellular peptides are not altered in response to elevated intracellular calcium, suggesting that calpain is not responsible for their production. The similarity of the peptidomes of the three cell lines and the lack of correlation with the predicted cellular degradome implies the selective formation or retention of these peptides, consistent with the hypothesis that they are functional in the cells. PMID:21204522

  3. The Importance of Interspecific Interactions on the Present Range of the Invasive Mosquito Aedes albopictus (Diptera: Culicidae) and Persistence of Resident Container Species in the United States.

    PubMed

    Fader, Joseph E

    2016-09-01

    Aedes albopictus (Skuse) established in the United States over 30 yr ago and quickly spread throughout the entire eastern half of the country. It has recently spread into western regions and projected climate change scenarios suggest continued expansion to the west and north. Aedes albopictus has had major impacts on, and been impacted by, a diverse array of resident mosquito species. Laying eggs at the edges of small, water-holding containers, hatched larvae develop within these containers feeding on detritus-based resources. Under limited resource conditions, Ae. albopictus has been shown to be a superior competitor to essentially all native and resident species in the United States. Adult males also mate interspecifically with at least one resident species with significant negative impacts on reproductive output for susceptible females. Despite these strong interference effects on sympatric species, competitor outcomes have been highly variable, ranging from outright local exclusion by Ae. albopictus, to apparent exclusion of Ae. albopictus in the presence of the same species. Context-dependent mechanisms that alter the relative strengths of inter- and intraspecific competition, as well as rapid evolution of satyrization-resistant females, may help explain these patterns of variable coexistence. Although there is a large body of research on interspecific interactions of Ae. albopictus in the United States, there remain substantial gaps in our understanding of the most important species interactions. Addressing these gaps is important in predicting the future distribution of this species and understanding consequences for resident species, including humans, that interact with this highly invasive mosquito. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  4. Characterization of endogenous calcium responses in neuronal cell lines.

    PubMed

    Vetter, Irina; Lewis, Richard J

    2010-03-15

    An increasing number of putative therapeutic targets have been identified in recent years for the treatment of neuronal pathophysiologies including pain, epilepsy, stroke and schizophrenia. Many of these targets signal through calcium (Ca(2+)), either by directly facilitating Ca(2+) influx through an ion channel, or through activation of G proteins that couple to intracellular Ca(2+) stores or voltage-gated Ca(2+) channels. Immortalized neuronal cell lines are widely used models to study neuropharmacology. However, systematic pharmacological characterization of the receptors and ion channels expressed in these cell lines is lacking. In this study, we systematically assessed endogenous Ca(2+) signaling in response to addition of agonists at potential therapeutic targets in a range of cell lines of neuronal origin (ND7/23, SH-SY5Y, 50B11, F11 and Neuro2A cells) as well as HEK293 cells, a cell line commonly used for over-expression of receptors and ion channels. This study revealed a remarkable diversity of endogenous Ca(2+) responses in these cell lines, with one or more cell lines responding to addition of trypsin, bradykinin, ATP, nicotine, acetylcholine, histamine and neurotensin. Subtype specificity of these responses was inferred from agonist potency and the effect of receptor subtype specific antagonist. Surprisingly, HEK293 and SH-SY5Y cells responded to the largest number of agonists with potential roles in neuronal signaling. These findings have implications for the heterologous expression of neuronal receptors and ion channels in these cell lines, and highlight the potential of neuron-derived cell lines for the study of a range of endogenously expressed receptors and ion channels that signal through Ca(2+). Crown Copyright 2009. Published by Elsevier Inc. All rights reserved.

  5. Toxic effect and genotoxicity of the semisynthetic derivatives dillapiole ethyl ether and dillapiole n-butyl ether for control of Aedes albopictus (Diptera: Culicidae).

    PubMed

    da Fonseca Meireles, Sabrina; Domingos, Pedro Rauel Cândido; da Silva Pinto, Ana Cristina; Rafael, Míriam Silva

    2016-09-01

    Two derivatives of dillapiole, dillapiole ethyl ether (1KL39-B) and butyl ether-n dillapiole (1KL43-C), were studied for their toxicity and genotoxicity against Aedes albopictus, to help develop new strategies for the control of this potential vector of dengue and other arboviruses, because it is resistant to synthetic insecticides. Eggs and larvae exposed to different concentrations of 1KL39-B (25, 30, 50, 70, and 80μg/mL) and of 1KL43-C (12.5, 20, 25, 30 and 40μg/mL) exhibited toxicity and susceptibility, with 100% mortality. The LC50 was 55.86±1.57μg/mL for 1KL39-B and 25.60±1.24μg/mL for 1KL43-C, while the LC90 was 70.12μg/mL for 1KL39-B and 41.51μg/mL for 1KL43-C. The gradual decrease in oviposition of the females of the G1 to G4 generations was proportional to the increase in concentrations of these compounds, which could be related to the cumulative effect of cell anomalies in neuroblasts and oocytes (P<0.05), including micronuclei, budding, multinucleated cells and nuclear bridges. These findings showed that both 1KL39-B and 1KL43-C can serve as potential alternatives in the control of A. albopictus. Copyright © 2016. Published by Elsevier B.V.

  6. [Establishment of human embryonic stem cell lines and their therapeutic application].

    PubMed

    Suemori, Hirofumi

    2004-03-01

    Embryonic stem (ES) cell lines are pluripotent stem cell lines that can be propagated indefinitely in culture, retaining their potency to differentiate into every type of cell and tissue in the body. ES cell lines were first established from mouse blastocysts, and have been used for research in developmental biology. ES cells have been proven to be very valuable in the genetic modification of the mouse, especially in producing knockout mice. Since establishment of human ES cell lines was reported, their use in cell replacement therapies has been enthusiastically expected. There have been reports of the differentiation of several useful cell types from human ES cell lines, and clinical use of functional tissues and cells from human ES cells is anticipated. In Japan, there have also been many demands for the use of human ES cells in basic and pre-clinical research. We obtained governmental permission to establish human ES cell lines in April 2002 and started research using donated frozen embryos in January 2003. We successfully established three ES cell line from three blastocysts. These cell lines will be distributed at cost to researchers who have governmental permission to use human ES cells.

  7. Molecular characterization of immortalized normal and dysplastic oral cell lines.

    PubMed

    Dickman, Christopher T D; Towle, Rebecca; Saini, Rajan; Garnis, Cathie

    2015-05-01

    Cell lines have been developed for modeling cancer and cancer progression. The molecular background of these cell lines is often unknown to those using them to model disease behaviors. As molecular alterations are the ultimate drivers of cell phenotypes, having an understanding of the molecular make-up of these systems is critical for understanding the disease biology modeled. Six immortalized normal, one immortalized dysplasia, one self-immortalized dysplasia, and two primary normal cell lines derived from oral tissues were analyzed for DNA copy number changes and changes in both mRNA and miRNA expression using SMRT-v.2 genome-wide tiling comparative genomic hybridization arrays, Agilent Whole Genome 4x44k expression arrays, and Exiqon V2.M-RT-PCR microRNA Human panels. DNA copy number alterations were detected in both normal and dysplastic immortalized cell lines-as well as in the single non-immortalized dysplastic cell line. These lines were found to have changes in expression of genes related to cell cycle control as well as alterations in miRNAs that are deregulated in clinical oral squamous cell carcinoma tissues. Immortal lines-whether normal or dysplastic-had increased disruption in expression relative to primary lines. All data are available as a public resource. Molecular profiling experiments have identified DNA, mRNA, and miRNA alterations for a panel of normal and dysplastic oral tissue cell lines. These data are a valuable resource to those modeling diseases of the oral mucosa, and give insight into the selection of model cell lines and the interpretation of data from those lines. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  8. Malignant hematopoietic cell lines: in vitro models for the study of natural killer cell leukemia-lymphoma.

    PubMed

    Drexler, H G; Matsuo, Y

    2000-05-01

    Malignancies involving natural killer (NK) cells are rare disorders. The complexity of NK cell-involving disorders has only recently been appreciated. Modern classifications discern immature (precursor) from mature NK cell leukemias-lymphomas. Continuous NK leukemia-lymphoma cell lines represent important model systems to study these neoplasms. While there are a number of putative NK cell lines which are, however, either not characterized, not immortalized, non-malignant, non-NK, or plain false cell lines, six bona fide malignant NK cell lines have been established and are sufficiently well characterized: HANK1, KHYG-1, NK-92, NKL, NK-YS and YT. Except for YT which was derived from a not further defined acute lymphoblastic lymphoma, these cell lines were established from patients with various NK cell malignancies. Five of the six cell lines are constitutively interleukin-2-dependent. Their immunoprofile is remarkably similar: CD1-, CD2+, surface CD3 (but cytoplasmic CD3epsilon+), CD4-, CD5-, CD7+, CD8-, CD16-, CD56+, CD57-, TCRalphabeta-, TCRgammadelta-, negative for B cell and myelomonocytic markers. The immunoglobulin heavy chain and T cell receptor genes are all in germline configuration. All six lines show complex chromosomal alterations, with both numerical and structural aberrations, attesting to their malignant and monoclonal nature. Functionally, these cells which contain azurophilic granules in their cytoplasm are nearly universally positive in NK activity assays. Three of five cell lines are Epstein-Barr virus-positive (type II latency). The composite data on these six cell lines allow for the operational definition of a typical malignant NK cell line profile. NK leukemia-lymphoma cell lines will prove invaluable for studies of normal and malignant NK cell biology.

  9. Establishment and characterization of feeder cell-dependent bovine fetal liver cell lines.

    PubMed

    Talbot, Neil C; Wang, Ling; Garrett, Wesley M; Caperna, Thomas J; Tang, Young

    2016-03-01

    The establishment and initial characterization of bovine fetal liver cell lines are described. Bovine fetal hepatocytes were cultured from the liver of a 34-d bovine fetus by physical disruption of the liver tissue. Released liver cells and clumps of cells were plated on STO (SIMS mouse strain, thioguanine- and ouabain-resistant) feeder layers and were cultured in a medium supplemented with 10% fetal bovine serum. After 2-3 wk, primary colonies of hepatocytes were observed by phase-contrast microscopic observation. Individual hepatocyte colonies were colony-cloned into independent bovine fetal liver (BFL) cell lines. Two cell lines, BFL-6 and BFL-9, grew the best of several isolates, and they were further characterized for growth potential and for hepatocyte morphology and function. The two cell lines were found to grow markedly better in the presence of the transforming growth factor (TGF)-beta inhibitor, SB431542 (1 μM). Their continuous culture also depended on a particular medium height-for T12.5 flasks, 3 ml total medium produced optimum growth. Higher or lower amounts of medium caused less cell growth or cessation of growth. The cell lines were propagated for over a year at split ratios of 1:2 or 1:3 at each passage until reaching senescence at approximately 30 passages. The cells were laterally polarized with well-developed canalicular spaces occurring between adjacent BFL cells. Treatment of the cultures with cyclic adenosine monophosphate (cAMP)-stimulating chemicals or peptides (e.g., forskolin or glucagon) caused physical expansion of the canaliculi between the cells within 15 min. The cells secreted a spectrum of serum proteins, were positive for the expression of several hepatocyte-specific genes, and converted ammonia to urea, although at a relatively low rate. The culture system provides an in vitro model of fetal bovine hepatocytes and is the first demonstration of the continuous culture of normal bovine hepatocytes as cell lines.

  10. The USDA-ARS area-wide project for management of the asian tiger mosquito, Aedes albopictus

    USDA-ARS?s Scientific Manuscript database

    The Asian tiger mosquito, Aedes albopictus, is among the most invasive species in the world. Established in the U.S. since 1985, this species now infests 30 states and continues to spread internationally. Concerned public health officials recognize this species as an important vector of chikunguny...

  11. Spatial distribution of Aedes aegypti and Aedes albopictus in relation to geo-ecological features in South Andaman, Andaman and Nicobar Islands, India.

    PubMed

    Shriram, A N; Sivan, A; Sugunan, A P

    2018-04-01

    The study was undertaken in South Andaman district, comprising three tehsils, viz. Port Blair, Ferrargunj and Little Andaman Tehsils, respectively. Intensive pupal infestation surveys were carried out along the National Highway (NH 223), the main passenger and trade route, referred to as Great Andaman Trunk Road. Sampling locations at every 3 km were geo-referenced with global positioning system unit. A total of 17314 water collections were examined from 29 locations across the South Andaman district, among which 1021 (5.9%) were colonized by immature stages of Aedes albopictus, Aedes aegypti and other mosquito species. Ae. aegypti were found in 12 locations, showing higher infestation in the densely built Aberdeen Bazaar. Breeding populations of Ae. albopictus were observed in 27 sampling locations. Both the species were not recorded in two Northern localities. In the areas where both the species are present, they were often found in the same developmental sites, suggesting convergent habitat selection. The most frequently encountered man-made, artificial and natural developmental sites were fixed cement tanks, plastic drums, plastic cans, metal drums, metal pots, discarded tires, coconut shells, leaf axils and tree holes. Ae. aegypti and Ae. albopictus were observed in varying proportions in Port Blair and Ferrargunj Tehsils, while the former species appeared to be absent in Little Andaman. This study elucidates the spatial distribution of Ae. aegypti and Ae. albopictus with preponderance of the latter species, pointing towards arboviral transmission and assumes public health importance in South Andaman district, endemic for dengue.

  12. Establishment of an ASPL-TFE3 renal cell carcinoma cell line (S-TFE).

    PubMed

    Hirobe, Megumi; Masumori, Naoya; Tanaka, Toshiaki; Kitamura, Hiroshi; Tsukamoto, Taiji

    2013-06-01

    Xp11 translocation renal cell carcinoma is a rare disease diagnosed in children and adolescents in the advanced stage with an aggressive clinical course. Various gene fusions including the transcription factor E3 (TFE3) gene located on chromosome X cause the tumor. We established an Xp11 translocation renal cell carcinoma cell line from a renal tumor in a 18-y-old Japanese female and named it "S-TFE." The cell line and its xenograft demonstrated definite gene fusion including TFE3. They showed strong nuclear staining for TFE3 in immunohistochemistry, TFE3 gene rearrangement in dual-color, break-apart FISH analysis and ASPL-TFE3 type 1 fusion transcripts detected by RT-PCR and direct DNA sequencing. Although many renal cell carcinoma cell lines have been established and investigated, only a few cell lines are recognized as Xp11.2 translocation carcinoma. S-TFE will be useful to examine the characteristics and drug susceptibility of Xp11 translocation renal cell carcinoma.

  13. Differential Susceptibilities of Aedes aegypti and Aedes albopictus from the Americas to Zika Virus

    PubMed Central

    Vazeille, Marie; Yebakima, André; Girod, Romain; Goindin, Daniella; Dupont-Rouzeyrol, Myrielle; Lourenço-de-Oliveira, Ricardo; Failloux, Anna-Bella

    2016-01-01

    Background Since the major outbreak in 2007 in the Yap Island, Zika virus (ZIKV) causing dengue-like syndromes has affected multiple islands of the South Pacific region. In May 2015, the virus was detected in Brazil and then spread through South and Central America. In December 2015, ZIKV was detected in French Guiana and Martinique. The aim of the study was to evaluate the vector competence of the mosquito spp. Aedes aegypti and Aedes albopictus from the Caribbean (Martinique, Guadeloupe), North America (southern United States), South America (Brazil, French Guiana) for the currently circulating Asian genotype of ZIKV isolated from a patient in April 2014 in New Caledonia. Methodology/Principal Findings Mosquitoes were orally exposed to an Asian genotype of ZIKV (NC-2014-5132). Upon exposure, engorged mosquitoes were maintained at 28°±1°C, a 16h:8h light:dark cycle and 80% humidity. 25–30 mosquitoes were processed at 4, 7 and 14 days post-infection (dpi). Mosquito bodies (thorax and abdomen), heads and saliva were analyzed to measure infection, dissemination and transmission, respectively. High infection but lower disseminated infection and transmission rates were observed for both Ae. aegypti and Ae. albopictus. Ae. aegypti populations from Guadeloupe and French Guiana exhibited a higher dissemination of ZIKV than the other Ae. aegypti populations examined. Transmission of ZIKV was observed in both mosquito species at 14 dpi but at a low level. Conclusions/Significance This study suggests that although susceptible to infection, Ae. aegypti and Ae. albopictus were unexpectedly low competent vectors for ZIKV. This may suggest that other factors such as the large naïve population for ZIKV and the high densities of human-biting mosquitoes contribute to the rapid spread of ZIKV during the current outbreak. PMID:26938868

  14. Genetic evidence for a worldwide chaotic dispersion pattern of the arbovirus vector, Aedes albopictus

    PubMed Central

    Manni, Mosè; Guglielmino, Carmela R.; Scolari, Francesca; Vega-Rúa, Anubis; Failloux, Anna-Bella; Somboon, Pradya; Lisa, Antonella; Savini, Grazia; Bonizzoni, Mariangela; Gomulski, Ludvik M.; Malacrida, Anna R.

    2017-01-01

    Background Invasive species represent a global concern for their rapid spread and the possibility of infectious disease transmission. This is the case of the global invader Aedes albopictus, the Asian tiger mosquito. This species is a vector of medically important arboviruses, notably chikungunya (CHIKV), dengue (DENV) and Zika (ZIKV). The reconstruction of the complex colonization pattern of this mosquito has great potential for mitigating its spread and, consequently, disease risks. Methodology/Principal findings Classical population genetics analyses and Approximate Bayesian Computation (ABC) approaches were combined to disentangle the demographic history of Aedes albopictus populations from representative countries in the Southeast Asian native range and in the recent and more recently colonized areas. In Southeast Asia, the low differentiation and the high co-ancestry values identified among China, Thailand and Japan indicate that, in the native range, these populations maintain high genetic connectivity, revealing their ancestral common origin. China appears to be the oldest population. Outside Southeast Asia, the invasion process in La Réunion, America and the Mediterranean Basin is primarily supported by a chaotic propagule distribution, which cooperates in maintaining a relatively high genetic diversity within the adventive populations. Conclusions/Significance From our data, it appears that independent and also trans-continental introductions of Ae. albopictus may have facilitated the rapid establishment of adventive populations through admixture of unrelated genomes. As a consequence, a great amount of intra-population variability has been detected, and it is likely that this variability may extend to the genetic mechanisms controlling vector competence. Thus, in the context of the invasion process of this mosquito, it is possible that both population ancestry and admixture contribute to create the conditions for the efficient transmission of

  15. Differential Susceptibilities of Aedes aegypti and Aedes albopictus from the Americas to Zika Virus.

    PubMed

    Chouin-Carneiro, Thais; Vega-Rua, Anubis; Vazeille, Marie; Yebakima, André; Girod, Romain; Goindin, Daniella; Dupont-Rouzeyrol, Myrielle; Lourenço-de-Oliveira, Ricardo; Failloux, Anna-Bella

    2016-03-01

    Since the major outbreak in 2007 in the Yap Island, Zika virus (ZIKV) causing dengue-like syndromes has affected multiple islands of the South Pacific region. In May 2015, the virus was detected in Brazil and then spread through South and Central America. In December 2015, ZIKV was detected in French Guiana and Martinique. The aim of the study was to evaluate the vector competence of the mosquito spp. Aedes aegypti and Aedes albopictus from the Caribbean (Martinique, Guadeloupe), North America (southern United States), South America (Brazil, French Guiana) for the currently circulating Asian genotype of ZIKV isolated from a patient in April 2014 in New Caledonia. Mosquitoes were orally exposed to an Asian genotype of ZIKV (NC-2014-5132). Upon exposure, engorged mosquitoes were maintained at 28° ± 1 °C, a 16h:8h light:dark cycle and 80% humidity. 25-30 mosquitoes were processed at 4, 7 and 14 days post-infection (dpi). Mosquito bodies (thorax and abdomen), heads and saliva were analyzed to measure infection, dissemination and transmission, respectively. High infection but lower disseminated infection and transmission rates were observed for both Ae. aegypti and Ae. albopictus. Ae. aegypti populations from Guadeloupe and French Guiana exhibited a higher dissemination of ZIKV than the other Ae. aegypti populations examined. Transmission of ZIKV was observed in both mosquito species at 14 dpi but at a low level. This study suggests that although susceptible to infection, Ae. aegypti and Ae. albopictus were unexpectedly low competent vectors for ZIKV. This may suggest that other factors such as the large naïve population for ZIKV and the high densities of human-biting mosquitoes contribute to the rapid spread of ZIKV during the current outbreak.

  16. Establishment of immortalized murine mesothelial cells and a novel mesothelioma cell line.

    PubMed

    Blum, Walter; Pecze, László; Felley-Bosco, Emanuela; Worthmüller-Rodriguez, Janine; Wu, Licun; Vrugt, Bart; de Perrot, Marc; Schwaller, Beat

    2015-08-01

    Mesothelial cells are susceptible to asbestos fiber-induced cytotoxicity and on longer time scales to transformation; the resulting mesothelioma is a highly aggressive neoplasm that is considered as incurable at the present time Zucali et al. (Cancer Treatment Reviews 37:543-558, 2011). Only few murine cell culture models of immortalized mesothelial cells and mesothelioma cell lines exist to date. We generated SV40-immortalized cell lines derived from wild-type (WT) and neurofibromatosis 2 (merlin) heterozygote (Nf2+/-) mice, both on a commonly used genetic background, C57Bl/6J. All immortalized mesothelial clones consistently grow in DMEM supplemented with fetal bovine serum. Cells can be passaged for more than 40 times without any signs of morphological changes or a decrease in proliferation rate. The tumor suppressor gene NF2 is one of the most frequently mutated genes in human mesothelioma, but its detailed function is still unknown. Thus, these genotypically distinct cell lines likely relevant for malignant mesothelioma formation are expected to serve as useful in vitro models, in particular to compare with in vivo studies in mice of the same genotype. Furthermore, we generated a novel murine mesothelioma cell line RN5 originating from an Nf2+/- mouse subjected to repeated crocidolite exposure. RN5 cells are highly tumorigenic.

  17. Casein gene expression in mouse mammary epithelial cell lines: Dependence upon extracellular matrix and cell type

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Medina, D.; Oborn, C.J.; Li, M.L.

    1987-09-01

    The COMMA-D mammary cell line exhibits mammary-specific functional differentiation under appropriate conditions in cell culture. The cytologically heterogeneous COMMA-D parental line and the clonal lines DB-1, TA-5, and FA-1 derived from the COMMA-D parent were examined for similar properties of functional differentiation. In monolayer cell culture, the cell lines DB-1, TA-5, FA-1, and MA-4 were examined for expression of mammary-specific and epithelial-specific proteins by an indirect immunofluorescence assay. The clonal cell lines were relatively homogeneous in their respective staining properties and seemed to represent three subpopulations found in the heterogeneous parental COMMA-D lines. None of the four clonal lines appearedmore » to represent myoepithelial cells. The cell lines were examined for expression of {beta}-casein mRNA in the presence or absence of prolactin. The inducibility of {beta}-casein in the COMMA-D cell line was further enhanced by a reconstituted basement membrane preparation enriched in laminin, collagen IV, and proteoglycans. These results support the hypothesis that the functional response of inducible mammary cell populations is a result of interaction among hormones, multiple extracellular matrix components, and specific cell types.« less

  18. The effect of temperature on the extrinsic incubation period and infection rate of dengue virus serotype 2 infection in Aedes albopictus.

    PubMed

    Xiao, Fang-Zhen; Zhang, Yi; Deng, Yan-Qin; He, Si; Xie, Han-Guo; Zhou, Xiao-Nong; Yan, Yan-Sheng

    2014-11-01

    Dengue fever is an acute mosquito-borne viral disease caused by dengue virus (DENV). Temperature may affect the efficiency of the mosquito vectors in spreading DENV. Aedes albopictus mosquitoes were infected orally with a DENV2 suspension and incubated at different temperatures. Subsequently, DENV2 antigen was collected from salivary gland and thorax-abdomen samples on different days postinfection and tested using an immunofluorescence assay to determine the extrinsic incubation period and infection rate. As the temperature increased, the extrinsic DENV2 incubation period in Ae. albopictus gradually shortened, and infection rates showed a tendency to initially increase, followed by a subsequent decrease.

  19. Surveillance and Control of Aedes albopictus in the Swiss-Italian Border Region: Differences in Egg Densities between Intervention and Non-intervention Areas

    PubMed Central

    Suter, Tobias T.; Flacio, Eleonora; Feijoó Fariña, Begoña; Engeler, Lukas; Tonolla, Mauro; Regis, Lêda N.; de Melo Santos, Maria A. V.; Müller, Pie

    2016-01-01

    Background Aedes albopictus, the Asian tiger mosquito, originates from the tropical and subtropical regions of Southeast Asia. Over the recent decades it has been passively spread across the globe, primarily through the used tyre trade and passive transportation along major traffic routes. A. albopictus is a proven vector for many arboviruses, most notably chikungunya and dengue, with recent outbreaks also in continental Europe. In southern Switzerland, in the Canton of Ticino A. albopictus was spotted for the first time in 2003. Since then the local authorities have implemented a control programme based on larval source reduction. Despite these efforts, mosquito densities have increased over the last decade, casting doubts on the effectiveness of such larval control programmes. Methodology/Principal Findings The Italian communities just across the Swiss-Italian border lack a control programme. This motivated us to compare the intervention and the non-intervention areas side by side in an attempt to find evidence for, or against, the effectiveness of larval A. albopictus control. Using ovitraps and a randomised sampling scheme, we examined the seasonal and spatial abundance of A. albopictus in sylvatic and urban environments across the Swiss-Italian border in 2012 and 2013. In the urban environments of the non-intervention area, egg densities were 2.26 times higher as compared to the intervention area. In the sylvatic environments, as compared to the urban environments, egg densities were 36% in the intervention area and 18% in the non-intervention area. Conclusions/Significance Though alternative explanations are also valid, the results support the hypothesis that the Ticino intervention programme does have an impact. At the same time the data also suggest that current larval interventions fall short in gaining full control over the mosquito, calling for the evaluation of additional, or alternative, approaches. Ideally, these should also consider inclusion of the

  20. Susceptibility profile and metabolic mechanisms involved in Aedes aegypti and Aedes albopictus resistant to DDT and deltamethrin in the Central African Republic.

    PubMed

    Ngoagouni, Carine; Kamgang, Basile; Brengues, Cécile; Yahouedo, Gildas; Paupy, Christophe; Nakouné, Emmanuel; Kazanji, Mirdad; Chandre, Fabrice

    2016-11-24

    Aedes aegypti and Ae. albopictus are the main epidemic vectors of dengue, chikungunya and Zika viruses worldwide. Their control during epidemics relies mainly on control of larvae and adults with insecticides. Unfortunately, loss of susceptibility of both species to several insecticide classes limits the efficacy of interventions. In Africa, where Aedes-borne viruses are of growing concern, few data are available on resistance to insecticides. To fill this gap, we assessed the susceptibility to insecticides of Ae. aegypti and Ae. albopictus populations in the Central African Republic (CAR) and studied the mechanisms of resistance. Immature stages were sampled between June and September 2014 in six locations in Bangui (the capital of CAR) for larval and adult bioassays according to WHO standard procedures. We also characterized DDT- and pyrethroid-resistant mosquitoes molecularly and biochemically, including tests for the activities of nonspecific esterases (α and β), mixed-function oxidases, insensitive acetylcholinesterase and glutathione S-transferases. Larval bioassays, carried out to determine the lethal concentrations (LC 50 and LC 95 ) and resistance ratios (RR 50 and RR 95 ), suggested that both vector species were susceptible to Bacillus thuringiensis var. israeliensis and to temephos. Bioassays of adults showed susceptibility to propoxur and fenitrothion, except for one Ae. albopictus population that was suspected to be resistant to fenithrothion. None of the Ae. aegypti populations was fully susceptible to DDT. Ae. albopictus presented a similar profile to Ae. aegypti but with a lower mortality rate (41%). Possible resistance to deltamethrin was observed among Ae. aegypti and Ae. albopictus, although some were susceptible. No kdr mutations were detected in either species; however, the activity of detoxifying enzymes was higher in most populations than in the susceptible Ae. aegypti strain, confirming decreased susceptibility to DDT and deltamethrin

  1. Trapping the Tiger: Efficacy of the Novel BG-Sentinel 2 With Several Attractants and Carbon Dioxide for Collecting Aedes albopictus (Diptera: Culicidae) in Southern France.

    PubMed

    Roiz, David; Duperier, Sandy; Roussel, Marion; Boussès, Philippe; Fontenille, Didier; Simard, Frédéric; Paupy, Christophe

    2016-03-01

    Targeted trapping of mosquito disease vectors plays an important role in the surveillance and control of mosquito-borne diseases. The Asian tiger mosquito, Aedes albopictus (Skuse), is an invasive species, which is spreading throughout the world, and is a potential vector of 24 arboviruses, particularly efficient in the transmission of chikungunya, dengue, and zika viruses. Using a 4 × 4 Latin square design, we assessed the efficacy of the new BG-Sentinel 2 mosquito trap using the attractants BG-lure and (R)-1-octen-3-ol cartridge, alone or in combination, and with and without carbon dioxide, for the field collection of Ae. albopictus mosquitoes.We found a synergistic effect of attractant and carbon dioxide that significantly increased twofold to fivefold the capture rate of Ae. albopictus. In combination with carbon dioxide, BG-lure cartridge is more effective than (R)-1-octen-3-ol in attracting females, while a combination of both attractants and carbon dioxide is the most effective for capturing males. In the absence of carbon dioxide, BG-lure cartridge alone did not increase the capture of males or females when compared with an unbaited trap. However, the synergistic effect of carbon dioxide and BG-lure makes this the most efficient combination in attracting Ae. albopictus.

  2. Temporal distribution and insecticide resistance profile of two major arbovirus vectors Aedes aegypti and Aedes albopictus in Yaoundé, the capital city of Cameroon.

    PubMed

    Kamgang, Basile; Yougang, Aurelie P; Tchoupo, Micareme; Riveron, Jacob M; Wondji, Charles

    2017-10-10

    Aedes aegypti and Ae. albopictus are the major epidemic vectors of several arbovirus diseases such as yellow fever, dengue, Zika and chikungunya worldwide. Both Aedes vectors are presents in Cameroon; however, knowledge on the dynamic of the distribution of these species across cities and their resistance profile to insecticide are limited. Here, we assessed the current distribution of Ae. aegypti and Ae. albopictus in Yaoundé, the Capital City, established the resistance profile to insecticides and explored the resistance mechanisms involved. Immature stages of Aedes were sampled in several breeding sites in December 2015 (dry season) and June 2016 (rainy season) in three central neighborhoods and four peripheral neighborhoods and reared to adult stage. The G0 adults were used for molecular identification and genotyping of F1534C mutation in Ae. aegypti. Bioassays and piperonyl butoxide (PBO) assays were carried out according to WHO guidelines. Analysis revealed that both species Ae. aegypti and Ae. albopictus are present in all prospected sites in Yaounde. However, in the dry season Ae. aegypti is most abundant in neighborhoods located in downtown. In contrast, Ae. albopictus was found most prevalent in suburbs whatever the season and in downtown during the rainy season. Bioassay analysis showed that both Ae. aegypti and Ae. albopictus, are resistant to 0.05% deltamethrin, 0.1% bendiocarb and 4% dichlorodiphenyltrichloroethane (DDT). A decreased of susceptibility to 0.75% permethrin and a full susceptibility to malathion 5% was observed. The mortality rate was increased after pre-exposure to synergist PBO. None of Ae. aegypti assayed revealed the presence of F1534C mutation. These findings are useful to planning vector control programme against arbovirus vectors in Cameroon and can be used as baseline in Africa where data on Aedes resistance is very scarce to plan further works.

  3. Micro-RNA expression in cisplatin resistant germ cell tumor cell lines

    PubMed Central

    2011-01-01

    Background We compared microRNA expression patterns in three cisplatin resistant sublines derived from paternal cisplatin sensitive germ cell tumor cell lines in order to improve our understanding of the mechanisms of cisplatin resistance. Methods Three cisplatin resistant sublines (NTERA-2-R, NCCIT-R, 2102EP-R) showing 2.7-11.3-fold increase in drug resistance after intermittent exposure to increasing doses of cisplatin were compared to their parental counterparts, three well established relatively cisplatin sensitive germ cell tumor cell lines (NTERA-2, NCCIT, 2102EP). Cells were cultured and total RNA was isolated from all 6 cell lines in three independent experiments. RNA was converted into cDNA and quantitative RT-PCR was run using 384 well low density arrays covering almost all (738) known microRNA species of human origin. Results Altogether 72 of 738 (9.8%) microRNAs appeared differentially expressed between sensitive and resistant cell line pairs (NTERA-2R/NTERA-2 = 43, NCCIT-R/NCCIT = 53, 2102EP-R/2102EP = 15) of which 46.7-95.3% were up-regulated (NTERA-2R/NTERA-2 = 95.3%, NCCIT-R/NCCIT = 62.3%, 2102EP-R/2102EP = 46.7%). The number of genes showing differential expression in more than one of the cell line pairs was 34 between NTERA-2R/NTERA-2 (79%) and NCCIT-R/NCCIT (64%), and 3 and 4, respectively, between these two cell lines and 2102EP-R/2102EP (about 27%). Only the has-miR-10b involved in breast cancer invasion and metastasis and has-miR-512-3p appeared to be up-regulated (2-3-fold) in all three cell lines. The hsa-miR-371-373 cluster (counteracting cellular senescence and linked with differentiation potency), as well as hsa-miR-520c/-520h (inhibiting the tumor suppressor p21) were 3.9-16.3 fold up-regulated in two of the three cisplatin resistant cell lines. Several new micro-RNA species missing an annotation towards cisplatin resistance could be identified. These were hsa-miR-512-3p/-515/-517/-518/-525 (up to 8.1-fold up-regulated) and hsa-miR-99a

  4. In vitro culture of human osteosarcoma cell lines: a comparison of functional characteristics for cell lines cultured in medium without and with fetal calf serum.

    PubMed

    Bruserud, Oystein; Tronstad, Karl Johan; Berge, Rolf

    2005-06-01

    Experimental in vitro models including well-characterised cell lines can be used to identify possible new therapeutic targets for the treatment of osteosarcoma. Culture media including inactivated serum is often recommended for in vitro culture of osteosarcoma cells, but the serum component then represents a nonstandardised parameter including a wide range of unidentified mediators. To improve the standardisation we have investigated whether serum-free culture media can be used in experimental in vitro studies of osteosarcoma cell lines. The seven osteosarcoma cell lines Cal72, SJSA-1, Saos-2, SK-ES-1, U2OS, 143.98.2, and KHOS-32IH were cultured in vitro in various serum-free media and media supplemented with 10% heat-inactivated fetal calf serum (FCS). Although proliferation often was relatively low in serum-free media (X-vivo 10, X-vivo 15, X-vivo 20, Stem Span SFEM), some cell lines (Cal72, KHOS-32IH, Saos-2) showed proliferation comparable with the recommended FCS-containing media even when using serum-free conditions. The optimal serum-free medium then varied between cell lines. We also compared 6 different FCS-containing media (including Stem Span with 10% FCS) and the optimal FCS-containing medium varied between cell lines. However, all cell lines proliferated well in Stem Span with FCS, and this medium was regarded as optimal for four of the lines. FCS could not be replaced by fatty acids or low density lipoprotein when testing the Stem Span medium. The release of a wide range of soluble mediators showed only minor differences when using serum-free and FCS-containing media (including Stem Span with and without FCS), and serum-free Stem Span could also be used for in vitro studies of mitogen-stimulated T cell activation in the presence of accessory osteosarcoma cells. The use of Stem Span with 10% FCS allowed the release of a wide range of chemokines by osteosarcoma cell lines (Cal72, SJSA-1), and the chemokine release profile was very similar to the

  5. Vector competence of Aedes albopictus and Aedes aegypti (Diptera: Culicidae) for the DEN2-FJ10 and DEN2-FJ11 strains of the dengue 2 virus in Fujian, China.

    PubMed

    Guo, Xiao-Xia; Li, Chun-Xiao; Zhang, Ying-Mei; Xing, Dan; Dong, Yan-De; Zhang, Heng-Duan; Qin, Cheng-Feng; Zhao, Tong-Yan

    2016-09-01

    Dengue is an acute, emerging, infectious disease transmitted by Aedes mosquitoes that has become a serious global public health problem. The DEN2-FJ10 and DEN2-FJ11 strains of the dengue 2 virus were originally isolated from the serum of a patient with dengue fever in Fujian Province, China, in 1999. Our data provide the first assessment of the vector competence of Aedes mosquitoes with respect to the DEN2-FJ10 and DEN2-FJ11 strains of the dengue virus. There were significant differences in the replication rates of these two viral strains in Aedes albopictus and Aedes aegypti (P<0.05); replication of the DEN2-FJ10 strain was greater in Ae. aegypti than in Ae. albopictus 5 days post infection whereas replication of the DEN2-FJ11 was greater in Ae. albopictus than in Ae. aegypti 7 days post infection. The replicative ability of the DEN2-FJ11 strain was greater than that of the DEN2-FJ10 strain in infected Ae. albopictus. In infected Ae. aegypti, rapid proliferation of the DEN2-FJ10 strain occurred earlier than in the DEN2-FJ11 strain. There were no significant differences in the midgut and salivary gland infection rates of Ae. albopictus and Ae. aegypti with respect to either viral strain. Although the DEN2-FJ10 and DEN2-FJ11 strains differ in their virulence to neonatal rats, there was no significant difference in the ability of either Ae. albopictus or Ae. aegypti to transmit the DEN2-FJ10 and DEN2-FJ10 strains of the dengue 2 virus (P>0.05). In summary, our results indicate that Ae. albopictus and Ae. aegypti mosquitoes are moderately competent vectors of the DEN2-FJ10 and DEN2-FJ11 strains of the dengue virus and provide the first evidence of the effect of these two viral strains on the vector competence of mosquitoes in China. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Adventitious viruses in insect cell lines used for recombinant protein expression.

    PubMed

    Geisler, Christoph; Jarvis, Donald L

    2018-04-01

    Insect cells are widely used for recombinant protein expression, typically as hosts for recombinant baculovirus vectors, but also for plasmid-mediated transient transfection or stable genetic transformation. Insect cells are used to express proteins for research, as well as to manufacture biologicals for human and veterinary medicine. Recently, several insect cell lines used for recombinant protein expression were found to be persistently infected with adventitious viruses. This has raised questions about how these infections might affect research performed using those cell lines. Furthermore, these findings raised serious concerns about the safety of biologicals produced using those cell lines. In response, new insect cell lines lacking adventitious viruses have been isolated for use as improved research tools and safer biological manufacturing platforms. Here, we review the scientific and patent literature on adventitious viruses found in insect cell lines, affected cell lines, and new virus-free cell lines. Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Effect of Larval Competition on Extrinsic Incubation Period and Vectorial Capacity of Aedes albopictus for Dengue Virus.

    PubMed

    Bara, Jeffrey; Rapti, Zoi; Cáceres, Carla E; Muturi, Ephantus J

    2015-01-01

    Despite the growing awareness that larval competition can influence adult mosquito life history traits including susceptibility to pathogens, the net effect of larval competition on human risk of exposure to mosquito-borne pathogens remains poorly understood. We examined how intraspecific larval competition affects dengue-2 virus (DENV-2) extrinsic incubation period and vectorial capacity of its natural vector Aedes albopictus. Adult Ae. albopictus from low and high-larval density conditions were orally challenged with DENV-2 and then assayed for virus infection and dissemination rates following a 6, 9, or 12-day incubation period using real-time quantitative reverse transcription PCR. We then modeled the effect of larval competition on vectorial capacity using parameter estimates obtained from peer-reviewed field and laboratory studies. Larval competition resulted in significantly longer development times, lower emergence rates, and smaller adults, but did not significantly affect the extrinsic incubation period of DENV-2 in Ae. albopictus. Our vectorial capacity models suggest that the effect of larval competition on adult mosquito longevity likely has a greater influence on vectorial capacity relative to any competition-induced changes in vector competence. Furthermore, we found that large increases in the viral dissemination rate may be necessary to compensate for small competition-induced reductions in daily survivorship. Our results indicate that mosquito populations that experience stress from larval competition are likely to have a reduced vectorial capacity, even when susceptibility to pathogens is enhanced.

  8. First report of field evolved resistance to agrochemicals in dengue mosquito, Aedes albopictus (Diptera: Culicidae), from Pakistan

    PubMed Central

    2011-01-01

    Background Agrochemicals have been widely used in Pakistan for several years. This exposes mosquito populations, particularly those present around agricultural settings, to an intense selection pressure for insecticide resistance. The aim of the present study was to investigate the toxicity of representative agrochemicals against various populations of Aedes albopictus (Skuse) collected from three different regions from 2008-2010. Results For organophosphates and pyrethroids, the resistance ratios compared with susceptible Lab-PK were in the range of 157-266 fold for chlorpyrifos, 24-52 fold for profenofos, 41-71 fold for triazofos, and 15-26 fold for cypermethrin, 15-53 fold for deltamethrin and 21-58 fold for lambdacyhalothrin. The resistance ratios for carbamates and new insecticides were in the range of 13-22 fold for methomyl, 24-30 fold for thiodicarb, and 41-101 fold for indoxacarb, 14-27 fold for emamectin benzoate and 23-50 fold for spinosad. Pair wise comparisons of the log LC50s of insecticides revealed correlation among several insecticides, suggesting a possible cross resistance mechanism. Moreover, resistance remained stable across 3 years, suggesting field selection for general fitness had also taken place for various populations of Ae. albopictus. Conclusion Moderate to high level of resistance to agrochemicals in Pakistani field populations of Ae. albopictus is reported here first time. The geographic extent of resistance is unknown but, if widespread, may lead to problems in future vector control. PMID:21781290

  9. Effect of Larval Competition on Extrinsic Incubation Period and Vectorial Capacity of Aedes albopictus for Dengue Virus

    PubMed Central

    Bara, Jeffrey; Rapti, Zoi; Cáceres, Carla E.; Muturi, Ephantus J.

    2015-01-01

    Despite the growing awareness that larval competition can influence adult mosquito life history traits including susceptibility to pathogens, the net effect of larval competition on human risk of exposure to mosquito-borne pathogens remains poorly understood. We examined how intraspecific larval competition affects dengue-2 virus (DENV-2) extrinsic incubation period and vectorial capacity of its natural vector Aedes albopictus. Adult Ae. albopictus from low and high-larval density conditions were orally challenged with DENV-2 and then assayed for virus infection and dissemination rates following a 6, 9, or 12-day incubation period using real-time quantitative reverse transcription PCR. We then modeled the effect of larval competition on vectorial capacity using parameter estimates obtained from peer-reviewed field and laboratory studies. Larval competition resulted in significantly longer development times, lower emergence rates, and smaller adults, but did not significantly affect the extrinsic incubation period of DENV-2 in Ae. albopictus. Our vectorial capacity models suggest that the effect of larval competition on adult mosquito longevity likely has a greater influence on vectorial capacity relative to any competition-induced changes in vector competence. Furthermore, we found that large increases in the viral dissemination rate may be necessary to compensate for small competition-induced reductions in daily survivorship. Our results indicate that mosquito populations that experience stress from larval competition are likely to have a reduced vectorial capacity, even when susceptibility to pathogens is enhanced. PMID:25951173

  10. Lung Cancer Cell Lines as Tools for Biomedical Discovery and Research

    PubMed Central

    Girard, Luc; Lockwood, William W.; Lam, Wan L.; Minna, John D.

    2010-01-01

    Lung cancer cell lines have made a substantial contribution to lung cancer translational research and biomedical discovery. A systematic approach to initiating and characterizing cell lines from small cell and non–small cell lung carcinomas has led to the current collection of more than 200 lung cancer cell lines, a number that exceeds those for other common epithelial cancers combined. The ready availability and widespread dissemination of the lines to investigators worldwide have resulted in more than 9000 citations, including multiple examples of important biomedical discoveries. The high (but not perfect) genomic similarities between lung cancer cell lines and the lung tumor type from which they were derived provide evidence of the relevance of their use. However, major problems including misidentification or cell line contamination remain. Ongoing studies and new approaches are expected to reveal the full potential of the lung cancer cell line panel. PMID:20679594

  11. Establishment of an ASPL-TFE3 renal cell carcinoma cell line (S-TFE)

    PubMed Central

    Hirobe, Megumi; Masumori, Naoya; Tanaka, Toshiaki; Kitamura, Hiroshi; Tsukamoto, Taiji

    2013-01-01

    Xp11 translocation renal cell carcinoma is a rare disease diagnosed in children and adolescents in the advanced stage with an aggressive clinical course. Various gene fusions including the transcription factor E3 (TFE3) gene located on chromosome X cause the tumor. We established an Xp11 translocation renal cell carcinoma cell line from a renal tumor in a 18-y-old Japanese female and named it “S-TFE.” The cell line and its xenograft demonstrated definite gene fusion including TFE3. They showed strong nuclear staining for TFE3 in immunohistochemistry, TFE3 gene rearrangement in dual-color, break-apart FISH analysis and ASPL-TFE3 type 1 fusion transcripts detected by RT-PCR and direct DNA sequencing. Although many renal cell carcinoma cell lines have been established and investigated, only a few cell lines are recognized as Xp11.2 translocation carcinoma. S-TFE will be useful to examine the characteristics and drug susceptibility of Xp11 translocation renal cell carcinoma. PMID:23760492

  12. Evaluation of pyriproxyfen dissemination via Aedes albopictus from a point source larvicide application in northeast Florida

    USDA-ARS?s Scientific Manuscript database

    The Asian tiger mosquito, Aedes albopictus, ranks among the most important vectors of dengue fever, Zika virus, and chikungunya virus. With no specific medications or vaccines available, vector control is the only way to combat these diseases. Autodissemination of the insect growth regulator pyripro...

  13. Apoptosis inhibitor of macrophage (AIM) reduces cell number in canine histiocytic sarcoma cell lines.

    PubMed

    Uchida, Mona; Saeki, Kohei; Maeda, Shingo; Tamahara, Satoshi; Yonezawa, Tomohiro; Matsuki, Naoaki

    2016-10-01

    Apoptosis inhibitor of macrophage (AIM) is initially reported to protect macrophages from apoptosis. In this study, we determined the effect of AIM on the macrophage-derived tumor, histiocytic sarcoma cell lines (HS) of dogs. Five HS and five other tumor cell lines were used. When recombinant canine AIM was applied to non-serum culture media, cell numbers of all the HS and two of other tumor cell lines decreased dose-dependently. The DNA fragmentation, TUNEL staining and flow cytometry tests revealed that AIM induced both of apoptosis and cell cycle arrest in the HS. Although AIM is known as an apoptosis inhibitor, these results suggest that a high dose of AIM could have an opposite function in HS and some tumor cell lines.

  14. Acrylamide affects proliferation and differentiation of the neural progenitor cell line C17.2 and the neuroblastoma cell line SH-SY5Y.

    PubMed

    Attoff, K; Kertika, D; Lundqvist, J; Oredsson, S; Forsby, A

    2016-09-01

    Acrylamide is a well-known neurotoxic compound and people get exposed to the compound by food consumption and environmental pollutants. Since acrylamide crosses the placenta barrier, the fetus is also being exposed resulting in a risk for developmental neurotoxicity. In this study, the neural progenitor cell line C17.2 and the neuroblastoma cell line SH-SY5Y were used to study proliferation and differentiation as alerting indicators for developmental neurotoxicity. For both cell lines, acrylamide reduced the number of viable cells by reducing proliferation and inducing cell death in undifferentiated cells. Acrylamide concentrations starting at 10fM attenuated the differentiation process in SH-SY5Y cells by sustaining cell proliferation and neurite outgrowth was reduced at concentrations from 10pM. Acrylamide significantly reduced the number of neurons starting at 1μM and altered the ratio between the different phenotypes in differentiating C17.2 cell cultures. Ten micromolar of acrylamide also reduced the expression of the neuronal and astrocyte biomarkers. Although the neurotoxic concentrations in the femtomolar range seem to be specific for the SH-SY5Y cell line, the fact that micromolar concentrations of acrylamide seem to attenuate the differentiation process in both cell lines raises the interest to further investigations on the possible developmental neurotoxicity of acrylamide. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  15. Larvicidal and repellent activity of the essential oil of Coriandrum sativum L. (Apiaceae) fruits against the filariasis vector Aedes albopictus Skuse (Diptera: Culicidae).

    PubMed

    Benelli, Giovanni; Flamini, Guido; Fiore, Giulia; Cioni, Pier Luigi; Conti, Barbara

    2013-03-01

    The essential oils of many Apiaceae species have been already studied for their insecticidal and repellent properties against insect pests. In this research, the essential oil (EO) extracted from the fruits of Coriandrum sativum L. (Apiaceae) was evaluated for the first time for its larvicidal and repellent activities against the most invasive mosquito worldwide, Aedes albopictus Skuse (Diptera: Culicidae). The chemical composition of C. sativum EO was investigated by gas chromatography with electron impact mass spectrometry analysis. Coriander EO was mainly composed by monoterpene hydrocarbons and oxygenated monoterpenes, with linalool (83.6 %) as the major constituent. C. sativum EO exerted toxic activity against A. albopictus larvae: LC(50) was 421 ppm, while LC(90) was 531.7 ppm. Repellence trials highlighted that C. sativum EO was a good repellent against A. albopictus, also at lower dosages: RD(50) was 0.0001565 μL/cm(2) of skin, while RD(90) was 0.002004 μL/cm(2). At the highest dosage (0.2 μL/cm(2) of skin), the protection time achieved with C. sativum essential oil was higher than 60 min. This study adds knowledge about the chemical composition of C. sativum EO as well as to the larvicidal and repellent activity exerted by this EO against A. albopictus. On this basis, we believe that our findings could be useful for the development of new and safer products against the Asian tiger mosquito.

  16. The Cytochrome P450 gene CYP6P12 confers pyrethroid resistance in kdr-free Malaysian populations of the dengue vector Aedes albopictus.

    PubMed

    Ishak, Intan H; Riveron, Jacob M; Ibrahim, Sulaiman S; Stott, Rob; Longbottom, Joshua; Irving, Helen; Wondji, Charles S

    2016-04-20

    Control of Aedes albopictus, major dengue and chikungunya vector, is threatened by growing cases of insecticide resistance. The mechanisms driving this resistance remain poorly characterised. This study investigated the molecular basis of insecticide resistance in Malaysian populations of Ae. albopictus. Microarray-based transcription profiling revealed that metabolic resistance (cytochrome P450 up-regulation) and possibly a reduced penetration mechanism (consistent over-expression of cuticular protein genes) were associated with pyrethroid resistance. CYP6P12 over-expression was strongly associated with pyrethroid resistance whereas CYP6N3 was rather consistently over-expressed across carbamate and DDT resistant populations. Other detoxification genes also up-regulated in permethrin resistant mosquitoes included a glucuronosyltransferase (AAEL014279-RA) and the glutathione-S transferases GSTS1 and GSTT3. Functional analyses further supported that CYP6P12 contributes to pyrethroid resistance in Ae. albopictus as transgenic expression of CYP6P12 in Drosophila was sufficient to confer pyrethroid resistance in these flies. Furthermore, molecular docking simulations predicted CYP6P12 possessing enzymatic activity towards pyrethroids. Patterns of polymorphism suggested early sign of selection acting on CYP6P12 but not on CYP6N3. The major role played by P450 in the absence of kdr mutations suggests that addition of the synergist PBO to pyrethroids could improve the efficacy of this insecticide class and overcome resistance in field populations of Ae. albopictus.

  17. Standard operating procedures for standardized mass rearing of the dengue and chikungunya vectors Aedes aegypti and Aedes albopictus (Diptera: Culicidae) - I - egg quantification.

    PubMed

    Zheng, Min-Lin; Zhang, Dong-Jing; Damiens, David D; Yamada, Hanano; Gilles, Jeremie R L

    2015-01-23

    Quantification of eggs prior to rearing the immature stages of mosquitoes is an essential step in establishing a standardized mass rearing system. To develop a simple and accurate method of egg quantification for Aedes aegypti and Aedes albopictus, the relationship between egg number and weight, as well as egg number and volume, were studied. Known quantities of eggs (1,000, 3,000, 6,000, 12,000, 15,000, 18,000, 21,000 and 27,000) were counted and subsequently their weight and volume were measured. Best-fit curves and regression equations were used to describe relationships between Aedes egg number and both weight and volume. Eighteen thousand Ae. aegypti eggs weighed 159.8 mg and had a volume of 277.4 μl, compared to measurements of 131.5 mg and 230.3 μl for Ae. albopictus. The eggs of Ae. aegypti were thus larger and heavier than those of Ae. albopictus. The use of weight and volume to quantify egg number was validated by counting volumes and weights of eggs expected to correspond to 3,000 and 18,000 eggs of each species; significant correlations were found in all cases except in the case of 3,000 Ae. albopictus eggs measured by volume. Methods for egg quantification were validated and shown to be a consistent and practical means to achieve uniform distribution of Aedes larvae between rearing trays, important for optimal mass rearing of the immature stages of Aedes mosquitoes.

  18. Identification of a novel rhabdovirus in Spodoptera frugiperda cell lines.

    PubMed

    Ma, Hailun; Galvin, Teresa A; Glasner, Dustin R; Shaheduzzaman, Syed; Khan, Arifa S

    2014-06-01

    The Sf9 cell line, derived from Spodoptera frugiperda, is used as a cell substrate for biological products, and no viruses have been reported in this cell line after extensive testing. We used degenerate PCR assays and massively parallel sequencing (MPS) to identify a novel RNA virus belonging to the order Mononegavirales in Sf9 cells. Sequence analysis of the assembled virus genome showed the presence of five open reading frames (ORFs) corresponding to the genes for the N, P, M, G, and L proteins in other rhabdoviruses and an unknown ORF of 111 amino acids located between the G- and L-protein genes. BLAST searches indicated that the S. frugiperda rhabdovirus (Sf-rhabdovirus) was related in a limited region of the L-protein gene to Taastrup virus, a newly discovered member of the Mononegavirales from a leafhopper (Hemiptera), and also to plant rhabdoviruses, particularly in the genus Cytorhabdovirus. Phylogenetic analysis of sequences in the L-protein gene indicated that Sf-rhabdovirus is a novel virus that branched with Taastrup virus. Rhabdovirus morphology was confirmed by transmission electron microscopy of filtered supernatant samples from Sf9 cells. Infectivity studies indicated potential transient infection by Sf-rhabdovirus in other insect cell lines, but there was no evidence of entry or virus replication in human cell lines. Sf-rhabdovirus sequences were also found in the Sf21 parental cell line of Sf9 cells but not in other insect cell lines, such as BT1-TN-5B1-4 (Tn5; High Five) cells and Schneider's Drosophila line 2 [D.Mel.(2); SL2] cells, indicating a species-specific infection. The results indicate that conventional methods may be complemented by state-of-the-art technologies with extensive bioinformatics analysis for identification of novel viruses. The Spodoptera frugiperda Sf9 cell line is used as a cell substrate for the development and manufacture of biological products. Extensive testing has not previously identified any viruses in this cell

  19. Identification of a Novel Rhabdovirus in Spodoptera frugiperda Cell Lines

    PubMed Central

    Ma, Hailun; Galvin, Teresa A.; Glasner, Dustin R.; Shaheduzzaman, Syed

    2014-01-01

    ABSTRACT The Sf9 cell line, derived from Spodoptera frugiperda, is used as a cell substrate for biological products, and no viruses have been reported in this cell line after extensive testing. We used degenerate PCR assays and massively parallel sequencing (MPS) to identify a novel RNA virus belonging to the order Mononegavirales in Sf9 cells. Sequence analysis of the assembled virus genome showed the presence of five open reading frames (ORFs) corresponding to the genes for the N, P, M, G, and L proteins in other rhabdoviruses and an unknown ORF of 111 amino acids located between the G- and L-protein genes. BLAST searches indicated that the S. frugiperda rhabdovirus (Sf-rhabdovirus) was related in a limited region of the L-protein gene to Taastrup virus, a newly discovered member of the Mononegavirales from a leafhopper (Hemiptera), and also to plant rhabdoviruses, particularly in the genus Cytorhabdovirus. Phylogenetic analysis of sequences in the L-protein gene indicated that Sf-rhabdovirus is a novel virus that branched with Taastrup virus. Rhabdovirus morphology was confirmed by transmission electron microscopy of filtered supernatant samples from Sf9 cells. Infectivity studies indicated potential transient infection by Sf-rhabdovirus in other insect cell lines, but there was no evidence of entry or virus replication in human cell lines. Sf-rhabdovirus sequences were also found in the Sf21 parental cell line of Sf9 cells but not in other insect cell lines, such as BT1-TN-5B1-4 (Tn5; High Five) cells and Schneider's Drosophila line 2 [D.Mel.(2); SL2] cells, indicating a species-specific infection. The results indicate that conventional methods may be complemented by state-of-the-art technologies with extensive bioinformatics analysis for identification of novel viruses. IMPORTANCE The Spodoptera frugiperda Sf9 cell line is used as a cell substrate for the development and manufacture of biological products. Extensive testing has not previously identified any

  20. Development of apoptosis-resistant dihydrofolate reductase-deficient Chinese hamster ovary cell line.

    PubMed

    Lee, Suk Kyoo; Lee, Gyun Min

    2003-06-30

    Apoptosis-resistant dihydrofolate reductase-deficient CHO cell line (dhfr(-) CHO-bcl2) was developed by introduction of the bcl-2 gene into the dhfr(-) CHO cell line (DUKX-B11, ATCC CRL-9096) and subsequent selection of clones stably overexpressing Bcl-2 in the absence of selection pressure. When the dhfr(-) CHO-bcl2 cell line was used as a host cell line for development of a recombinant CHO (rCHO) cell line expressing a humanized antibody, it displayed stable expression of the bcl-2 gene during rCHO cell line development and no detrimental effect of Bcl-2 overexpression on specific antibody productivity. Taken together, the results obtained demonstrate that the use of an apoptosis-resistant dhfr(-) CHO cell line as the host cell line saves the effort of establishing an apoptosis-resistant rCHO cell line and expedites the development process of apoptosis-resistant rCHO cells producing therapeutic proteins. Copyright 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 872-876, 2003.

  1. CRISPR/Cas9 mediated generation of stable chondrocyte cell lines with targeted gene knockouts; analysis of an aggrecan knockout cell line.

    PubMed

    Yang, Maozhou; Zhang, Liang; Stevens, Jeff; Gibson, Gary

    2014-12-01

    The Swarm rat chondrosarcoma (RCS) cell lines derived from a spontaneous neoplasm in a rat spine several decades ago have provided excellent models of chondrosarcoma tumor development. In addition the robust chondrocyte phenotype (expression of a large panel of genes identical to that seen in normal rat cartilage) and the ability to generate cell clones have facilitated their extensive use in the identification of chondrocyte proteins and genes. The clustered regularly interspersed short palindromic repeat (CRISPR) technology employing the RNA-guided nuclease Cas9 has rapidly dominated the genome engineering field as a unique and powerful gene editing tool. We have generated a stable RCS cell line (RCS Cas9) expressing the nuclease Cas9 that enables the editing of any target gene or non-coding RNA by simple transfection with a guide RNA. As proof of principle, stable cell lines with targeted ablation of aggrecan expression (Acan KO) were generated and characterized. The studies show that stable chondrocyte cell lines with targeted genome editing can be quickly generated from RCS Cas9 cells using this system. The Acan KO cell lines also provided a tool for characterizing the response of chondrocytes to aggrecan loss and the role of aggrecan in chondrosarcoma development. Loss of aggrecan expression while not affecting the chondrocyte phenotype resulted in a much firmer attachment of cells to their substrate in culture. Large changes in the expression of several genes were observed in response to the absence of the proteoglycan matrix, including those for several small leucine rich proteoglycans (SLRPs), transcription factors and membrane transporters. Acan KO cells failed to form a substantial chondrosarcoma when injected subcutaneously in nude mice consistent with previous suggestions that the glycosaminoglycan-rich matrix surrounding the chondrosarcoma protects it from destruction by the host immune system. The studies provide new understanding of aggrecan

  2. Phenotypes and Karyotypes of Human Malignant Mesothelioma Cell Lines

    PubMed Central

    Relan, Vandana; Morrison, Leanne; Parsonson, Kylie; Clarke, Belinda E.; Duhig, Edwina E.; Windsor, Morgan N.; Matar, Kevin S.; Naidoo, Rishendran; Passmore, Linda; McCaul, Elizabeth; Courtney, Deborah; Yang, Ian A.; Fong, Kwun M.; Bowman, Rayleen V.

    2013-01-01

    Background Malignant mesothelioma is an aggressive tumour of serosal surfaces most commonly pleura. Characterised cell lines represent a valuable tool to study the biology of mesothelioma. The aim of this study was to develop and biologically characterise six malignant mesothelioma cell lines to evaluate their potential as models of human malignant mesothelioma. Methods Five lines were initiated from pleural biopsies, and one from pleural effusion of patients with histologically proven malignant mesothelioma. Mesothelial origin was assessed by standard morphology, Transmission Electron Microscopy (TEM) and immunocytochemistry. Growth characteristics were assayed using population doubling times. Spectral karyotyping was performed to assess chromosomal abnormalities. Authentication of donor specific derivation was undertaken by DNA fingerprinting using a panel of SNPs. Results Most of cell lines exhibited spindle cell shape, with some retaining stellate shapes. At passage 2 to 6 all lines stained positively for calretinin and cytokeratin 19, and demonstrated capacity for anchorage-independent growth. At passage 4 to 16, doubling times ranged from 30–72 hours, and on spectral karyotyping all lines exhibited numerical chromosomal abnormalities ranging from 41 to 113. Monosomy of chromosomes 8, 14, 22 or 17 was observed in three lines. One line displayed four different karyotypes at passage 8, but only one karyotype at passage 42, and another displayed polyploidy at passage 40 which was not present at early passages. At passages 5–17, TEM showed characteristic features of mesothelioma ultrastructure in all lines including microvilli and tight intercellular junctions. Conclusion These six cell lines exhibit varying cell morphology, a range of doubling times, and show diverse passage-dependent structural chromosomal changes observed in malignant tumours. However they retain characteristic immunocytochemical protein expression profiles of mesothelioma during

  3. Characteristics of cell lines established from human colorectal carcinoma.

    PubMed

    Park, J G; Oie, H K; Sugarbaker, P H; Henslee, J G; Chen, T R; Johnson, B E; Gazdar, A

    1987-12-15

    We have characterized 14 human colorectal carcinoma cell lines established from primary and metastatic sites by us during the years 1982 to 1985. Five lines were established in fully defined ACL-4 medium and 9 in serum supplemented R10 medium. However, after establishment, cultures could be grown interchangeably in either medium. The lines grew as floating cell aggregates in ACL-4 medium, while most demonstrated substrate adherence in R10 medium. The lines had relatively long doubling times and low cloning efficiencies. Twelve were tumorigenic in athymic nude mice when injected s.c., and two grew i.p. as well. Based on culture, xenograft, and ultrastructural morphologies, the 14 lines could be subtyped as follows: 4 were well differentiated; 5 were moderately differentiated; 4 were poorly differentiated; and 1 was a mucinous carcinoma. Membrane associated antigens characteristic for gastrointestinal cells (carcinoembryonic antigen, CA 19-9, and TAG-72 antigens) were expressed by 50-71% of the lines. Lines expressing carcinoembryonic antigen and CA 19-9 actively secreted these antigens into the supernatant fluids while TAG-72 antigen was not secreted. Surprisingly, 5 of 7 of the original tumor samples tested and 13 of 14 cultured lines expressed L-dopa decarboxylase activity, which is a characteristic enzyme marker of neuroendocrine cells and tumors. In addition, one poorly differentiated cell line contained dense core granules, characteristic of endocrine secretion. Preliminary cytogenetic analyses indicated that 9 of 11 lines examined contained double minute chromosomes. In addition, 3 of the 9 lines with double minutes also had homogeneously staining regions. These findings indicate a high incidence of amplification of one or more as yet unidentified genes.

  4. Picking Cell Lines for High-Throughput Transcriptomic Toxicity ...

    EPA Pesticide Factsheets

    High throughput, whole genome transcriptomic profiling is a promising approach to comprehensively evaluate chemicals for potential biological effects. To be useful for in vitro toxicity screening, gene expression must be quantified in a set of representative cell types that captures the diversity of potential responses across chemicals. The ideal dataset to select these cell types would consist of hundreds of cell types treated with thousands of chemicals, but does not yet exist. However, basal gene expression data may be useful as a surrogate for representing the relevant biological space necessary for cell type selection. The goal of this study was to identify a small (< 20) number of cell types that capture a large, quantifiable fraction of basal gene expression diversity. Three publicly available collections of Affymetrix U133+2.0 cellular gene expression data were used: 1) 59 cell lines from the NCI60 set; 2) 303 primary cell types from the Mabbott et al (2013) expression atlas; and 3) 1036 cell lines from the Cancer Cell Line Encyclopedia. The data were RMA normalized, log-transformed, and the probe sets mapped to HUGO gene identifiers. The results showed that <20 cell lines capture only a small fraction of the total diversity in basal gene expression when evaluated using either the entire set of 20960 HUGO genes or a subset of druggable genes likely to be chemical targets. The fraction of the total gene expression variation explained was consistent when

  5. Effects of desiccation stress on adult female longevity in Aedes aegypti and Ae. albopictus (Diptera: Culicidae): results of a systematic review and pooled survival analysis.

    PubMed

    Schmidt, Chris A; Comeau, Genevieve; Monaghan, Andrew J; Williamson, Daniel J; Ernst, Kacey C

    2018-04-25

    Transmission dynamics of mosquito-borne viruses such as dengue, Zika and chikungunya are affected by the longevity of the adult female mosquito. Environmental conditions influence the survival of adult female Aedes mosquitoes, the primary vectors of these viruses. While the association of temperature with Aedes mortality has been relatively well-explored, the role of humidity is less established. The current study's goals were to compile knowledge of the influence of humidity on adult survival in the important vector species Aedes aegypti and Ae. albopictus, and to quantify this relationship while accounting for the modifying effect of temperature. We performed a systematic literature review to identify studies reporting experimental results informing the relationships among temperature, humidity and adult survival in Ae. aegypti and Ae. albopictus. Using a novel simulation approach to harmonize disparate survival data, we conducted pooled survival analyses via stratified and mixed effects Cox regression to estimate temperature-dependent associations between humidity and mortality risk for these species across a broad range of temperatures and vapor pressure deficits. After screening 1517 articles, 17 studies (one in semi-field and 16 in laboratory settings) met inclusion criteria and collectively reported results for 192 survival experiments. We review and synthesize relevant findings from these studies. Our stratified model estimated a strong temperature-dependent association of humidity with mortality in both species, though associations were not significant for Ae. albopictus in the mixed effects model. Lowest mortality risks were estimated around 27.5 °C and 21.5 °C for Ae. aegypti and Ae. albopictus, respectively, and mortality increased non-linearly with decreasing humidity. Aedes aegypti had a survival advantage relative to Ae. albopictus in the stratified model under most conditions, but species differences were not significant in the mixed effects model

  6. Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties

    PubMed Central

    Gudjonsson, Thorarinn; Villadsen, René; Nielsen, Helga Lind; Rønnov-Jessen, Lone; Bissell, Mina J.; Petersen, Ole William

    2002-01-01

    The epithelial compartment of the human breast comprises two distinct lineages: the luminal epithelial and the myoepithelial lineage. We have shown previously that a subset of the luminal epithelial cells could convert to myoepithelial cells in culture signifying the possible existence of a progenitor cell. We therefore set out to identify and isolate the putative precursor in the luminal epithelial compartment. Using cell surface markers and immunomagnetic sorting, we isolated two luminal epithelial cell populations from primary cultures of reduction mammoplasties. The major population coexpresses sialomucin (MUC+) and epithelial-specific antigen (ESA+) whereas the minor population has a suprabasal position and expresses epithelial specific antigen but no sialomucin (MUC−/ESA+). Two cell lines were further established by transduction of the E6/E7 genes from human papilloma virus type 16. Both cell lines maintained a luminal epithelial phenotype as evidenced by expression of the tight junction proteins, claudin-1 and occludin, and by generation of a high transepithelial electrical resistance on semipermeable filters. Whereas in clonal cultures, the MUC+/ESA+ epithelial cell line was luminal epithelial restricted in its differentiation repertoire, the suprabasal-derived MUC−/ESA+ epithelial cell line was able to generate itself as well as MUC+/ESA+ epithelial cells and Thy-1+/α-smooth muscle actin+ (ASMA+) myoepithelial cells. The MUC−/ESA+ epithelial cell line further differed from the MUC+/ESA+ epithelial cell line by the expression of keratin K19, a feature of a subpopulation of epithelial cells in terminal duct lobular units in vivo. Within a reconstituted basement membrane, the MUC+/ESA+ epithelial cell line formed acinus-like spheres. In contrast, the MUC−/ESA+ epithelial cell line formed elaborate branching structures resembling uncultured terminal duct lobular units both by morphology and marker expression. Similar structures were obtained by

  7. Establishment and characterization of Macaca fascicularis lymphoblastoid cell lines.

    PubMed

    Manning, C H; Heise, E R

    1992-01-01

    A panel of cynomolgus macaque lymphoblastoid cell lines (LCL) was established by transforming peripheral blood mononuclear cells (PBMC) with Herpesvirus papio (HVP), and selected lines were examined by flow cytometry. Results indicate that HVP-transformed macaque LCL are phenotypically heterogeneous and resemble human Epstein-Barr virus (EBV)-transformed LCL in the abundant expression of major histocompatibility complex (MHC) class I and class II molecules. At least some lines are of B cell origin.

  8. Molecular characterization of breast cancer cell lines through multiple omic approaches.

    PubMed

    Smith, Shari E; Mellor, Paul; Ward, Alison K; Kendall, Stephanie; McDonald, Megan; Vizeacoumar, Frederick S; Vizeacoumar, Franco J; Napper, Scott; Anderson, Deborah H

    2017-06-05

    Breast cancer cell lines are frequently used as model systems to study the cellular properties and biology of breast cancer. Our objective was to characterize a large, commonly employed panel of breast cancer cell lines obtained from the American Type Culture Collection (ATCC 30-4500 K) to enable researchers to make more informed decisions in selecting cell lines for specific studies. Information about these cell lines was obtained from a wide variety of sources. In addition, new information about cellular pathways that are activated within each cell line was generated. We determined key protein expression data using immunoblot analyses. In addition, two analyses on serum-starved cells were carried out to identify cellular proteins and pathways that are activated in these cells. These analyses were performed using a commercial PathScan array and a novel and more extensive phosphopeptide-based kinome analysis that queries 1290 phosphorylation events in major signaling pathways. Data about this panel of breast cancer cell lines was also accessed from several online sources, compiled and summarized for the following areas: molecular classification, mRNA expression, mutational status of key proteins and other possible cancer-associated mutations, and the tumorigenic and metastatic capacity in mouse xenograft models of breast cancer. The cell lines that were characterized included 10 estrogen receptor (ER)-positive, 12 human epidermal growth factor receptor 2 (HER2)-amplified and 18 triple negative breast cancer cell lines, in addition to 4 non-tumorigenic breast cell lines. Within each subtype, there was significant genetic heterogeneity that could impact both the selection of model cell lines and the interpretation of the results obtained. To capture the net activation of key signaling pathways as a result of these mutational combinations, profiled pathway activation status was examined. This provided further clarity for which cell lines were particularly deregulated

  9. Authentication of the R06E Fruit Bat Cell Line

    PubMed Central

    Jordan, Ingo; Munster, Vincent J.; Sandig, Volker

    2012-01-01

    Fruit bats and insectivorous bats are believed to provide a natural reservoir for a wide variety of infectious diseases. Several lines of evidence, including the successful isolation of infectious viruses, indicate that Marburg virus and Ravn virus have found a major reservoir in colonies of the Egyptian rousette (Rousettus aegyptiacus). To facilitate molecular studies on virus-reservoir host interactions and isolation of viruses from environmental samples, we established cell lines from primary cells of this animal. The cell lines were given to several laboratories until we realized that a contamination with Vero cells in one of the cultures had occurred. Here we describe a general diagnostic procedure for identification of cross-species contamination with the focus on Vero and Rousettus cell lines, and summarize newly discovered properties of the cell lines that may pertain to pathogen discovery. PMID:22754654

  10. Authentication of the R06E fruit bat cell line.

    PubMed

    Jordan, Ingo; Munster, Vincent J; Sandig, Volker

    2012-05-01

    Fruit bats and insectivorous bats are believed to provide a natural reservoir for a wide variety of infectious diseases. Several lines of evidence, including the successful isolation of infectious viruses, indicate that Marburg virus and Ravn virus have found a major reservoir in colonies of the Egyptian rousette (Rousettus aegyptiacus). To facilitate molecular studies on virus-reservoir host interactions and isolation of viruses from environmental samples, we established cell lines from primary cells of this animal. The cell lines were given to several laboratories until we realized that a contamination with Vero cells in one of the cultures had occurred. Here we describe a general diagnostic procedure for identification of cross-species contamination with the focus on Vero and Rousettus cell lines, and summarize newly discovered properties of the cell lines that may pertain to pathogen discovery.

  11. Selection of Phage Display Peptides Targeting Human Pluripotent Stem Cell-Derived Progenitor Cell Lines.

    PubMed

    Bignone, Paola A; Krupa, Rachel A; West, Michael D; Larocca, David

    2016-01-01

    The ability of human pluripotent stem cells (hPS) to both self-renew and differentiate into virtually any cell type makes them a promising source of cells for cell-based regenerative therapies. However, stem cell identity, purity, and scalability remain formidable challenges that need to be overcome for translation of pluripotent stem cell research into clinical applications. Directed differentiation from hPS cells is inefficient and residual contamination with pluripotent cells that have the potential to form tumors remains problematic. The derivation of scalable (self-renewing) embryonic progenitor stem cell lines offers a solution because they are well defined and clonally pure. Clonally pure progenitor stem cell lines also provide a means for identifying cell surface targeting reagents that are useful for identification, tracking, and repeated derivation of the corresponding progenitor stem cell types from additional hPS cell sources. Such stem cell targeting reagents can then be applied to the manufacture of genetically diverse banks of human embryonic progenitor cell lines for drug screening, disease modeling, and cell therapy. Here we present methods to identify human embryonic progenitor stem cell targeting peptides by selection of phage display libraries on clonal embryonic progenitor cell lines and demonstrate their use for targeting quantum dots (Qdots) for stem cell labeling.

  12. Large genetic differentiation and low variation in vector competence for dengue and yellow fever viruses of Aedes albopictus from Brazil, the United States, and the Cayman Islands.

    PubMed

    Lourenço de Oliveira, Ricardo; Vazeille, Marie; de Filippis, Ana Maria Bispo; Failloux, Anna-Bella

    2003-07-01

    We conducted a population genetic analysis of Aedes albopictus collected from 20 sites in Brazil, the United States (Florida, Georgia, and Illinois), and the Cayman Islands. Using isoenzyme analysis, we examined genetic diversity and patterns of gene flow. High genetic differentiation was found among Brazilian samples, and between them and North American samples. Regression analysis of genetic differentiation according to geographic distances indicated that Ae. albopictus samples from Florida were genetically isolated by distance. Infection rates with dengue and yellow fever viruses showed greater differences between two Brazilian samples than between the two North American samples or between a Brazilian sample and a North American sample. Introductions and establishments of new Ae. albopictus populations in the Americas are still in progress, shaping population genetic composition and potentially modifying both dengue and yellow fever transmission patterns.

  13. Establishment of an immortal chicken embryo liver-derived cell line.

    PubMed

    Lee, Jeongyoon; Foster, Douglas N; Bottje, Walter G; Jang, Hyeon-Min; Chandra, Yohanna G; Gentles, Lauren E; Kong, Byung-Whi

    2013-06-01

    A continuously growing immortal cell substrate can be used for virus propagation, diagnostic purposes, and vaccine production. The aim of this study was to develop an immortal chicken cell line for efficient propagation of avian infectious viruses. From the various chicken embryo cells that were tested for life span extension, an immortalized chicken embryo liver (CEL) cell line, named CEL-im, was derived spontaneously without either oncogenic viruses or carcinogenic chemical treatment. Currently, CEL-im cells are growing 0.8 to 1.1 population doublings per day and have reached 120 passages. The CEL-im cell line is permissive for poultry infectious viruses, including avian metapneumovirus (AMPV), Marek's disease virus serotype 1 (MDV-1), and infectious laryngotracheitis virus. The CEL-im cells produced high AMPV titer (>10(5) pfu/mL), whereas very low titers (~10 pfu/mL) for MDV-1 and infectious laryngotracheitis virus were produced. To identify genetic alterations in the immortal CEL-im cell line, telomerase activity and mRNA expression for major cell cycle regulatory genes were determined during the immortalizing process. The CEL-im cell line has negative telomerase activity, and when compared with the primary passage 2 CEL cell counterpart, mRNA expression of tumor suppressor protein p53, mouse double minute 2 (Mdm2), cyclin dependent kinase (CDK) inhibitor p21 (p21(WAF)), and CDK inhibitor p16 (p16(INK4)) were downregulated in the CEL-im cell line, whereas retinoblastoma (Rb), transcription factor E2F, member 1 (E2F-1), and alternative reading frame of p16(INK4) (ARF) were upregulated. These results are similar to genetic alterations found previously in immortal chicken embryo fibroblast (CEF) cell lines that showed efficient propagation of MDV-1. Therefore, this newly established CEL-im cell line can serve as an alternative cell substrate for the propagation of poultry viruses, such as AMPV.

  14. Characterization of three new serous epithelial ovarian cancer cell lines

    PubMed Central

    Ouellet, Véronique; Zietarska, Magdalena; Portelance, Lise; Lafontaine, Julie; Madore, Jason; Puiffe, Marie-Line; Arcand, Suzanna L; Shen, Zhen; Hébert, Josée; Tonin, Patricia N; Provencher, Diane M; Mes-Masson, Anne-Marie

    2008-01-01

    Background Cell lines constitute a powerful model to study cancer, and here we describe three new epithelial ovarian cancer (EOC) cell lines derived from poorly differentiated serous solid tumors (TOV-1946, and TOV-2223G), as well as the matched ascites for one case (OV-1946). Methods In addition to growth parameters, the cell lines were characterized for anchorage independent growth, migration and invasion potential, ability to form spheroids and xenografts in SCID mice. Results While all cell lines were capable of anchorage independent growth, only the TOV-1946 and OV-1946 cell lines were able to form spheroid and produce tumors. Profiling of keratins, p53 and Her2 protein expression was assessed by immunohistochemistry and western blot analyses. Somatic TP53 mutations were found in all cell lines, with TOV-1946 and OV-1946 harboring the same mutation, and none harbored the commonly observed somatic mutations in BRAF, KRAS or germline BRCA1/2 mutations found to recur in the French Canadian population. Conventional cytogenetics and spectral karyotype (SKY) analyses revealed complex karyotypes often observed in ovarian disease. Conclusion This is the first report of the establishment of matched EOC cell lines derived from both solid tumor and ascites of the same patient. PMID:18507860

  15. Establishment of a medium-scale mosquito facility: tests on mass production cages for Aedes albopictus (Diptera: Culicidae).

    PubMed

    Zhang, Dongjing; Li, Yongjun; Sun, Qiang; Zheng, Xiaoying; Gilles, Jeremie R L; Yamada, Hanano; Wu, Zhongdao; Xi, Zhiyong; Wu, Yu

    2018-03-19

    Mass egg production is an important component of Aedes albopictus mosquito control programs, such as the sterile insect technique and incompatible insect technique, which requires the releases of large number of sterile males. Developing standard operating procedures and optimized cages for adult maintenance of Ae. albopictus can improve the mass rearing efficiency. Three different sex ratios of females to males with a total number of 4,000 mosquitoes were tested by evaluating the insemination rate, egg production (total number of eggs per cage), female fecundity and egg hatch rate in small cage (30 × 30 × 30 cm). Blood meals with adenosine triphosphate (ATP, 0.05 g/ml), cage structures (Big cage A: 90 × 30 × 30 cm; Big cage B: 90 × 30 × 50 cm or 90 × 50 × 30 cm) and rearing densities (12,000, 16,000 and 20,000 mosquitoes, corresponding to 0.9 cm 2 /mosquito, 0.675 cm 2 /mosquito and 0.54 cm 2 /mosquito, respectively) were also tested and evaluated on the basis of egg production, female fecundity and egg hatch rate. An adult rearing unit holding 15 of Big cage A with optimal egg production was designed to produce 10 million eggs per rearing cycle in a 1.8 m 2 space. Female to male ratios at 3:1 in small cages resulted in higher egg production but did not affect insemination rate, female fecundity and egg hatch rate. A concentration of 0.05 g/ml of ATP added to blood meals improved the blood-feeding frequency and thus increased the overall egg production per cage. Cage structures affected the egg production per cage, but not egg hatch rate. A medium rearing density at 0.675 cm 2 /mosquito (16,000 mosquitoes) resulted in higher egg production compared to both low and high densities. An adult rearing unit for Ae. albopictus on the basis of Big cage A has been developed with the capacity of producing 10 million eggs within 15 days. Our results have indicated that the adult rearing methods and adult maintenance unit are recommended for Ae. albopictus mass

  16. Mapping Past, Present, and Future Climatic Suitability for Invasive Aedes Aegypti and Aedes Albopictus in the United States: A Process-Based Modeling Approach Using CMIP5 Downscaled Climate Scenarios

    NASA Technical Reports Server (NTRS)

    Donnelly, Marisa Anne Pella; Marcantonio, Matteo; Melton, Forrest S.; Barker, Christopher M.

    2016-01-01

    The ongoing spread of the mosquitoes, Aedes aegypti and Aedes albopictus, in the continental United States leaves new areas at risk for local transmission of dengue, chikungunya, and Zika viruses. All three viruses have caused major disease outbreaks in the Americas with infected travelers returning regularly to the U.S. The expanding range of these mosquitoes raises questions about whether recent spread has been enabled by climate change or other anthropogenic influences. In this analysis, we used downscaled climate scenarios from the NASA Earth Exchange Global Daily Downscaled Projections (NEX GDDP) dataset to model Ae. aegypti and Ae. albopictus population growth rates across the United States. We used a stage-structured matrix population model to understand past and present climatic suitability for these vectors, and to project future suitability under CMIP5 climate change scenarios. Our results indicate that much of the southern U.S. is suitable for both Ae. aegypti and Ae. albopictus year-round. In addition, a large proportion of the U.S. is seasonally suitable for mosquito population growth, creating the potential for periodic incursions into new areas. Changes in climatic suitability in recent decades for Ae. aegypti and Ae. albopictus have occurred already in many regions of the U.S., and model projections of future climate suggest that climate change will continue to reshape the range of Ae. aegypti and Ae. albopictus in the U.S., and potentially the risk of the viruses they transmit.

  17. Mapping past, present, and future climatic suitability for invasive Aedes aegypti and Aedes albopictus in the United States: a process-based modeling approach using CMIP5 downscaled climate scenarios

    NASA Astrophysics Data System (ADS)

    Donnelly, M. A. P.; Marcantonio, M.; Melton, F. S.; Barker, C. M.

    2016-12-01

    The ongoing spread of the mosquitoes, Aedes aegypti and Aedes albopictus, in the continental United States leaves new areas at risk for local transmission of dengue, chikungunya, and Zika viruses. All three viruses have caused major disease outbreaks in the Americas with infected travelers returning regularly to the U.S. The expanding range of these mosquitoes raises questions about whether recent spread has been enabled by climate change or other anthropogenic influences. In this analysis, we used downscaled climate scenarios from the NASA Earth Exchange Global Daily Downscaled Projections (NEX GDDP) dataset to model Ae. aegypti and Ae. albopictus population growth rates across the United States. We used a stage-structured matrix population model to understand past and present climatic suitability for these vectors, and to project future suitability under CMIP5 climate change scenarios. Our results indicate that much of the southern U.S. is suitable for both Ae. aegypti and Ae. albopictus year-round. In addition, a large proportion of the U.S. is seasonally suitable for mosquito population growth, creating the potential for periodic incursions into new areas. Changes in climatic suitability in recent decades for Ae. aegypti and Ae. albopictus have occurred already in many regions of the U.S., and model projections of future climate suggest that climate change will continue to reshape the range of Ae. aegypti and Ae. albopictus in the U.S., and potentially the risk of the viruses they transmit.

  18. Evaluation of Lambda-Cyhalothrin and Pyriproxyfen Barrier Treatments for Aedes albopictus (Diptera: Culicidae) Management in Urbanized Areas of New Jersey.

    PubMed

    Unlu, Isik; Williams, Gregory M; Rochlin, Ilia; Suman, Devi; Wang, Yi; Chandel, Kshitij; Gaugler, Randy

    2018-02-28

    Mosquito control programs in the United States are still searching for best management practices to control the Asian tiger mosquito, Aedes albopictus (Skuse; Diptera: Culicidae). Most intervention methods for this species are either labor intensive (e.g., source reduction) or short-term (e.g., ultra-low-volume adulticiding). We investigated the effectiveness of barrier spray pesticide applications within urban and suburban residential yards in New Jersey as a control strategy using a before-after-control-impact (BACI) approach. Applications of Demand CSR pyrethroid (9.7% AI lambda-cyhalothrin) only or combined Demand CSR and Archer IGR insect growth regulator (1.3% AI pyriproxyfen) applications resulted in significant and similar decreases in adult mosquito abundance post-treatment ranging from 78 to 74% respectively, compared with the untreated control. Both insecticides exceeded the 70% reduction threshold considered as effective for Ae. albopictus control for 2 to 4 wk. However, applications of Archer IGR alone did not reduce adult mosquito abundance. The field study results were supported by laboratory no-choice bioassays using treated leaf foliage. Our study is the first data driven evidence of the residual efficacy of barrier pesticide applications in New Jersey with lambda-cyhalothrin that provided significant reductions in adult Ae. albopictus populations for an extended duration.

  19. Peroxisomal abnormalities in the immortalized human hepatocyte (IHH) cell line.

    PubMed

    Klouwer, Femke C C; Koster, Janet; Ferdinandusse, Sacha; Waterham, Hans R

    2017-04-01

    The immortalized human hepatocyte (IHH) cell line is increasingly used for studies related to liver metabolism, including hepatic glucose, lipid, lipoprotein and triglyceride metabolism, and the effect of therapeutic interventions. To determine whether the IHH cell line is a good model to investigate hepatic peroxisomal metabolism, we measured several peroxisomal parameters in IHH cells and, for comparison, HepG2 cells and primary skin fibroblasts. This revealed a marked plasmalogen deficiency and a deficient fatty acid α-oxidation in the IHH cells, due to a defect of PEX7, a cytosolic receptor protein required for peroxisomal import of a subset of peroxisomal proteins. These abnormalities have consequences for the lipid homeostasis of these cells and thus should be taken into account for the interpretation of data previously generated by using this cell line and when considering using this cell line for future research.

  20. A comparative analysis of resistance testing methods in Aedes albopictus (Diptera: Culicidae) from St. Johns County, Florida

    USDA-ARS?s Scientific Manuscript database

    Aedes albopictus Skuse (Diptera: Culicidae) was tested for resistance to permethrin, bifenthrin, and malathion using Centers for Disease Control and Prevention (CDC) bottle bioassays and topical toxicology assays on adults and larval bioassays. Eggs were collected from 3 locations across St. Johns C...

  1. Macrophage cell lines derived from major histocompatibility complex II-negative mice

    NASA Technical Reports Server (NTRS)

    Beharka, A. A.; Armstrong, J. W.; Chapes, S. K.; Spooner, B. S. (Principal Investigator)

    1998-01-01

    Two bone-marrow-derived macrophage cell lines, C2D and C2Dt, were isolated from major histocompatibility class II negative knock-out mice. The C2D cell line was stabilized by continuous culture in colony-stimulating factor-1 and the C2Dt cell line was transformed with SV40 virus large T antigen. These cells exhibited phenotypic properties of macrophages including morphology and expression of Mac 1 and Mac 2 cell surface molecules. These cells also had comparable growth to the bone-marrow-derived macrophage cell line B6MP102. These new cell lines were not spontaneously cytotoxic and were only capable of modest killing of F5b tumor cells when stimulated with LPS and interferon-gamma, but not when stimulated with LPS alone or with staphylococcal exotoxin. C2D and C2Dt cells phagocytosed labeled Staphylococcus aureus similarly to B6MP102 cells but less well than C2D peritoneal macrophages. These cell lines secreted interleukin-6, but not tumor necrosis factor or nitric oxide in response to LPS or staphlococcal enterotoxins A or B C2D(t) cells were tumorigenic in C2D and C57BL/6J mice but C2D cells were not. These data suggest that macrophage cell lines can be established from bone marrow cells of major histocompatibility complex II-negative mice.

  2. Cell Line Modeling to Study Biomarker Panel in Prostate Cancer

    PubMed Central

    NickKholgh, Bita; Fang, Xiaolan; Winters, Shira M.; Raina, Anvi; Pandya, Komal S.; Gyabaah, Kenneth; Fino, Nora; Balaji, K.C.

    2016-01-01

    BACKGROUND African–American men with prostate cancer (PCa) present with higher-grade and -stage tumors compared to Caucasians. While the disparity may result from multiple factors, a biological basis is often strongly suspected. Currently, few well-characterized experimental model systems are available to study the biological basis of racial disparity in PCa. We report a validated in vitro cell line model system that could be used for the purpose. METHODS We assembled a PCa cell line model that included currently available African–American PCa cell lines and LNCaP (androgen-dependent) and C4-2 (castration-resistant) Caucasian PCa cells. The utility of the cell lines in studying the biological basis of variance in a malignant phenotype was explored using a multiplex biomarker panel consisting of proteins that have been proven to play a role in the progression of PCa. The panel expression was evaluated by Western blot and RT-PCR in cell lines and validated in human PCa tissues by RT-PCR. As proof-of-principle to demonstrate the utility of our model in functional studies, we performed MTS viability assays and molecular studies. RESULTS The dysregulation of the multiplex biomarker panel in primary African–American cell line (E006AA) was similar to metastatic Caucasian cell lines, which would suggest that the cell line model could be used to study an inherent aggressive phenotype in African–American men with PCa. We had previously demonstrated that Protein kinase D1 (PKD1) is a novel kinase that is down regulated in advanced prostate cancer. We established the functional relevance by over expressing PKD1, which resulted in decreased proliferation and epithelial mesenchymal transition (EMT) in PCa cells. Moreover, we established the feasibility of studying the expression of the multiplex biomarker panel in archived human PCa tissue from African–Americans and Caucasians as a prelude to future translational studies. CONCLUSION We have characterized a novel in

  3. Truck-mounted area-wide application of pyriproxyfen targeting Aedes aegypti and Aedes albopictus in northeast Florida

    USDA-ARS?s Scientific Manuscript database

    This study was conducted to determine the efficacy of truck-mounted ULV applications of pyriproxyfen against Aedes aegypti larvae in artificial water containers and wild adult Ae. albopictus populations in an urban setting. The study was conducted over a 3 ½ month period (Jun – Oct 2012), during wh...

  4. Widespread evidence for interspecific mating between Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in nature.

    PubMed

    Bargielowski, I E; Lounibos, L P; Shin, D; Smartt, C T; Carrasquilla, M C; Henry, A; Navarro, J C; Paupy, C; Dennett, J A

    2015-12-01

    Aedes aegypti and Aedes albopictus, two important vectors of the dengue and chikungunya viruses to humans, often come in contact in their invasive ranges. In these circumstances, a number of factors are thought to influence their population dynamics, including resource competition among the larval stages, prevailing environmental conditions and reproductive interference in the form of satyrization. As the distribution and abundance of Ae. aegypti and Ae. albopictus have profound epidemiological implications, understanding the competitive interactions that influence these patterns in nature is important. While evidence for resource competition and environmental factors had been gathered from the field, the evidence for reproductive interference, though strongly inferred through laboratory trials, remained sparse (one small-scale field trial). In this paper we demonstrate that low rates (1.12-3.73%) of interspecific mating occur in nature among populations of these species that have co-existed sympatrically from 3 to 150yrs. Finally this report contributes a new species-specific primer set for identifying the paternity of sperm extracted from field collected specimens. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Morphological study of the TK cholangiocarcinoma cell line with three-dimensional cell culture.

    PubMed

    Akiyoshi, Kohei; Kamada, Minori; Akiyama, Nobutake; Suzuki, Masafumi; Watanabe, Michiko; Fujioka, Kouki; Ikeda, Keiichi; Mizuno, Shuichi; Manome, Yoshinobu

    2014-04-01

    Cholangiocarcinoma is an intractable carcinoma originating from the bile duct epithelium. To gain an understanding of the cell biology of cholangiocarcinoma, in vitro cell culture is valuable. However, well‑characterized cell lines are limited. In the present study, the morphology of the TK cholangiocarcinoma cell line was analyzed by three‑dimensional culture. Dispersed TK cells were injected into a gelatin mesh scaffold and cultivated for 3‑20 days. The morphology of the TK cells was investigated by phase‑contrast microscopy, optical microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). TK cells were observed to proliferate three-dimensionally in the scaffold. The cells exhibited a globoid structure and attached to the scaffold. The SEM observation demonstrated typical microvilli and plicae on the surface of the structure. Light microscopy and TEM confirmed intercellular and cell‑to‑scaffold attachment in the three‑dimensional mesh. The culture also exhibited the formation of a duct-like structure covered by structured microvilli. In conclusion, three‑dimensional culture of TK cells demonstrated the morphological characteristics of cholangiocarcinoma in vitro. Production of high levels of carbohydrate antigen (CA)19‑9, CA50 and carcinoembryonic antigen was previously confirmed in the TK cell line. As a characteristic morphology was demonstrated in the present study, the TK cholangiocarcinoma cell line may be useful as an experimental model for further study of cholangiocarcinoma.

  6. Improving the efficiency of CHO cell line generation using glutamine synthetase gene knockout cells.

    PubMed

    Fan, Lianchun; Kadura, Ibrahim; Krebs, Lara E; Hatfield, Christopher C; Shaw, Margaret M; Frye, Christopher C

    2012-04-01

    Although Chinese hamster ovary (CHO) cells, with their unique characteristics, have become a major workhorse for the manufacture of therapeutic recombinant proteins, one of the major challenges in CHO cell line generation (CLG) is how to efficiently identify those rare, high-producing clones among a large population of low- and non-productive clones. It is not unusual that several hundred individual clones need to be screened for the identification of a commercial clonal cell line with acceptable productivity and growth profile making the cell line appropriate for commercial application. This inefficiency makes the process of CLG both time consuming and laborious. Currently, there are two main CHO expression systems, dihydrofolate reductase (DHFR)-based methotrexate (MTX) selection and glutamine synthetase (GS)-based methionine sulfoximine (MSX) selection, that have been in wide industrial use. Since selection of recombinant cell lines in the GS-CHO system is based on the balance between the expression of the GS gene introduced by the expression plasmid and the addition of the GS inhibitor, L-MSX, the expression of GS from the endogenous GS gene in parental CHOK1SV cells will likely interfere with the selection process. To study endogenous GS expression's potential impact on selection efficiency, GS-knockout CHOK1SV cell lines were generated using the zinc finger nuclease (ZFN) technology designed to specifically target the endogenous CHO GS gene. The high efficiency (∼2%) of bi-allelic modification on the CHO GS gene supports the unique advantages of the ZFN technology, especially in CHO cells. GS enzyme function disruption was confirmed by the observation of glutamine-dependent growth of all GS-knockout cell lines. Full evaluation of the GS-knockout cell lines in a standard industrial cell culture process was performed. Bulk culture productivity improved two- to three-fold through the use of GS-knockout cells as parent cells. The selection stringency was

  7. Host cell reactivation of gamma-irradiated adenovirus 5 in human cell lines of varying radiosensitivity.

    PubMed Central

    Eady, J. J.; Peacock, J. H.; McMillan, T. J.

    1992-01-01

    DNA repair processes play an important role in the determination of radiation response in both normal and tumour cells. We have investigated one aspect of DNA repair in a number of human cell lines of varying radiosensitivity using the adenovirus 5 host cell reactivation assay (HCR). In this technique, gamma-irradiated virions are used to infect cells and the ability of the cellular repair systems to process this damage is assayed by a convenient immunoperoxidase method recognising viral structural antigen expression on the cell membrane 48 h after infection. Reduced HCR was exhibited by radioresistant HeLa cells and by a radiosensitive neuroblastoma cell line, HX142. In contrast, an ataxia telangiectasia cell line, AT5 BIVA, did not show reduced HCR. On the basis of these results we can make no general conclusions about the relevance of HCR to cellular radiosensitivity. We have extended these studies to determine whether our cell lines exhibited enhanced viral reactivation (ER) following a small priming dose of gamma-radiation given to the cells before viral infection. No evidence for this phenomenon was found either in normal or tumour cell lines. PMID:1637659

  8. Subcloning of three osteoblastic cell lines with distinct differentiation phenotypes from the mouse osteoblastic cell line KS-4.

    PubMed

    Yamashita, T; Ishii, H; Shimoda, K; Sampath, T K; Katagiri, T; Wada, M; Osawa, T; Suda, T

    1996-11-01

    Three distinct osteoblastic cell lines (KS418, KS460, and KS483) were subcloned from the mouse osteoblastic KS-4 cells, which possessed the abilities not only to differentiate into mature osteoblasts, but also to support osteoclast differentiation in coculture with spleen cells. The order of the magnitude of the basal alkaline phosphatase (ALP) activity was KS483 > KS418 > KS460. KS483 cells were also more differentiated than KS418 and KS460 in terms of ALP activity and osteocalcin production, when cultured in growth medium containing 10% fetal bovine serum. In long-term culture, KS418 and KS483 apparently differentiated into mature osteoblasts and formed calcified nodules without addition of beta-glycerophosphate. Electron microscopic analysis demonstrated that calcification occurring in the nodules was initiated in the matrix vesicles as observed in bone formation in vivo. Nodule formation and mineral deposition occurred simultaneously in the presence of beta-glycerophosphate, but the former always preceded the latter without addition of beta-glycerophosphate. In contrast, KS460 cells did not show time-dependent increases of ALP activity, type I collagen expression and osteocalcin production, which were induced by treatment with recombinant osteogenic protein-1 (OP-1). The three cell lines similarly supported osteoclast differentiation in coculture with spleen cells in response to 1,25-dihydroxyvitamin D3. These results indicate that the three cell lines subcloned from the original KS-4 cells represent phenotypically distinct osteoblasts during osteoblast differentiation, but are equipped similarly with the capacity to support osteoclast differentiation. The subcloned cells of the KS-4 series may provide useful systems in which to study osteoblast differentiation and function.

  9. The effect of photoperiod on life history and blood-feeding activity in Aedes albopictus and Aedes aegypti (Diptera: Culicidae).

    PubMed

    Costanzo, K S; Schelble, S; Jerz, K; Keenan, M

    2015-06-01

    Several studies have examined how climatic variables such as temperature and precipitation may affect life history traits in mosquitoes that are important to disease transmission. Despite its importance as a seasonal cue in nature, studies investigating the influence of photoperiod on such traits are relatively few. This study aims to investigate how photoperiod alters life history traits, survival, and blood-feeding activity in Aedes albopictus (Skuse) and Aedes aegypti (Linnaeus). We performed three experiments that tested the effects of day length on female survival, development time, adult size, fecundity, adult life span, and propensity to blood feed in Ae. albopictus and Ae. aegypti. Each experiment had three photoperiod treatments: 1) short-day (10L:14D), 2) control (12L:12D), and 3) long-day (14L:10D). Aedes albopictus adult females were consistently larger in size when reared in short-day conditions. Aedes aegypti adult females from short-day treatments lived longer and were more likely to take a blood meal compared to other treatments. We discuss how species-specific responses may reflect alternative strategies evolved to increase survival during unfavorable conditions. We review the potential impacts of these responses on seasonal transmission patterns, such as potentially increasing vectorial capacity of Ae. aegypti during periods of shorter day lengths. © 2015 The Society for Vector Ecology.

  10. Derivation and Osmotolerance Characterization of Three Immortalized Tilapia (Oreochromis mossambicus) Cell Lines

    PubMed Central

    Gardell, Alison M.; Qin, Qin; Rice, Robert H.; Li, Johnathan; Kültz, Dietmar

    2014-01-01

    Fish cell cultures are becoming more widely used models for investigating molecular mechanisms of physiological response to environmental challenge. In this study, we derived two immortalized Mozambique tilapia (Oreochromis mossambicus) cell lines from brain (OmB) and lip epithelium (OmL), and compared them to a previously immortalized bulbus arteriosus (TmB) cell line. The OmB and OmL cell lines were generated without or with Rho-associated kinase (ROCK) inhibitor/3T3 feeder layer supplementation. Although both approaches were successful, ROCK inhibitor/feeder layer supplementation was found to offer the advantages of selecting for epithelial-like cell type and decreasing time to immortalization. After immortalization (≥ passage 5), we characterized the proteomes of the newly derived cell lines (OmB and OmL) using LCMS and identified several unique cell markers for each line. Subsequently, osmotolerance for each of the three cell lines following acute exposure to elevated sodium chloride was evaluated. The acute maximum osmotolerance of these tilapia cell lines (>700 mOsm/kg) was markedly higher than that of any other known vertebrate cell line, but was significantly higher in the epithelial-like OmL cell line. To validate the physiological relevance of these tilapia cell lines, we quantified the effects of acute hyperosmotic challenge (450 mOsm/kg and 700 mOsm/kg) on the transcriptional regulation of two enzymes involved in biosynthesis of the compatible organic osmolyte, myo-inositol. Both enzymes were found to be robustly upregulated in all three tilapia cell lines. Therefore, the newly established tilapia cells lines represent valuable tools for studying molecular mechanisms involved in the osmotic stress response of euryhaline fish. PMID:24797371

  11. Larvicidal and repellent activity of Hyptis suaveolens (Lamiaceae) essential oil against the mosquito Aedes albopictus Skuse (Diptera: Culicidae).

    PubMed

    Conti, Barbara; Benelli, Giovanni; Flamini, Guido; Cioni, Pier Luigi; Profeti, Raffaele; Ceccarini, Lucia; Macchia, Mario; Canale, Angelo

    2012-05-01

    Lamiaceae have traditionally been used in developing countries for their insecticidal and repellent properties against several insect species. In our research, the essential oil (EO) extracted from fresh leaves of Hyptis suaveolens (Lamiaceae), and its main constituents were evaluated for larvicidal and repellent activity against the Asian tiger mosquito, Aedes albopictus Skuse (Diptera: Culicidae), currently the most invasive mosquito worldwide. H. suaveolens EO had insecticidal activity against A. albopictus larvae and mortality was dosage dependent. At the highest dosages of 450 and 400 ppm, there were no significant differences on larval mortality, as mortality ranged between 98.33% and 93.33%, respectively. At dosages ranging from 250 to 350 ppm, mortality rates were lower and not significantly different from each other. Terpinolene was found to be the most effective pure compound. Efficacy protection from H. suaveolens EO, at dosages ranging from 0.03748 to 0.7496 μg cm(-2) of skin, was evaluated during 150 min of observation. Results indicated that this EO had a significant repellent activity (RD(50) = 0.00035 μg cm(-2); RD(90) = 0.00048 μg cm(-2)), with differences in repellency rates, as a function of both concentration and observation time. Protection time ranged from 16 to 135 min. These results clearly evidenced that the larvicidal and repellent activity of H. suaveolens EO could be used for the development of new and safer products against A. albopictus.

  12. Astaxanthin Inhibits Proliferation of Human Gastric Cancer Cell Lines by Interrupting Cell Cycle Progression.

    PubMed

    Kim, Jung Ha; Park, Jong-Jae; Lee, Beom Jae; Joo, Moon Kyung; Chun, Hoon Jai; Lee, Sang Woo; Bak, Young-Tae

    2016-05-23

    Astaxanthin is a carotenoid pigment that has antioxidant, antitumoral, and anti-inflammatory properties. In this in vitro study, we investigated the mechanism of anticancer effects of astaxanthin in gastric carcinoma cell lines. The human gastric adenocarcinoma cell lines AGS, KATO-III, MKN-45, and SNU-1 were treated with various concentrations of astaxanthin. A cell viability test, cell cycle analysis, and immunoblotting were performed. The viability of each cancer cell line was suppressed by astaxanthin in a dose-dependent manner with significantly decreased proliferation in KATO-III and SNU-1 cells. Astaxanthin increased the number of cells in the G0/G1 phase but reduced the proportion of S phase KATO-III and SNU-1 cells. Phosphorylated extracellular signal-regulated kinase (ERK) was decreased in an inverse dose-dependent correlation with astaxanthin concentration, and the expression of p27(kip-1) increased the KATO-III and SNU-1 cell lines in an astaxanthin dose-dependent manner. Astaxanthin inhibits proliferation by interrupting cell cycle progression in KATO-III and SNU-1 gastric cancer cells. This may be caused by the inhibition of the phosphorylation of ERK and the enhanced expression of p27(kip-1).

  13. Dose-Dependent Behavioral Response of the Mosquito Aedes albopictus to Floral Odorous Compounds

    PubMed Central

    Hao, Huiling; Sun, Jingcheng; Dai, Jianqing

    2013-01-01

    The value of using plant volatiles as attractants for trapping and spatial repellents to protect hosts against mosquitoes has been widely recognized. The current study characterized behavioral responses of Aedes albopictus (Skuse) (Diptera: Culicidae) to different concentrations, ranging from 6 to 96%, of several common floral odorous compounds, including linalool, geraniol, citronellal, eugenol, anisaldehyde, and citral, using a wind tunnel olfactometer system. The results indicated that female mosquitoes reacted differently to different concentrations of the tested compounds, and the reactions also were different when those chemicals were tested alone or in the presence of human host odor. When tested alone, anisaldehyde was attractive at all tested concentrations, eugenol was attractive only at concentrations of 48–96%, while citronellal, linalool, citral, and geraniol were attractive at lower concentrations and repellent at higher concentrations. When tested in the presence of a human host, all compounds except for anisaldehyde at all tested concentrations showed host-seeking inhibition to certain degrees. Based on the results, it was concluded that anisaldehyde was effective in attracting Ae. albopictus when used alone but could also remarkably inhibit the host-seeking ability at a concentration of 96%, while citral, geraniol, linalool, citronellal, and eugenol are suitable as spatial repellents. PMID:24779928

  14. Cell lines derived from the squash bug, Anasa tristis (Coreidae: Hemiptera).

    PubMed

    Goodman, Cynthia L; Ringbauer, Joseph A; Li, Yao-Fa; Lincoln, Tamra Reall; Stanley, David

    2017-05-01

    The squash bug, Anasa tristis, is a pest of cucurbits that exerts direct damage on crops and is a vector of plant pathogens. We established cell lines from this insect to serve as tools for basic biology, including virology and immunology, as well as applied studies, such as insecticide development programs. We initiated 15 cell cultures, using nine media or combinations of media. The media yielding the best results were a modification of Kimura's medium and a combination of two commercially available cell culture media (EX-CELL 420 and L15). We designated the two cell lines as BCIRL-AtE-CLG11 and BCIRL-AtE-CLG15. From the AtE-CLG15 line, we isolated two sub-lines, A and B. Of these, the most consistently replicating line was AtE-CLG15A. We determined the doubling time of this line (190 h) and its mean cell diameter (14.5 ± 0.7 μm). We characterized the AtE-CLG15A line using DAF-PCR. The BCIRL-AtE-CLG15A cell line is now available for researchers world-wide.

  15. Electrophysiological characteristics of embryonic stem cell-derived cardiomyocytes are cell line-dependent.

    PubMed

    Hannes, Tobias; Wolff, Marie; Doss, Michael Xavier; Pfannkuche, Kurt; Haustein, Moritz; Müller-Ehmsen, Jochen; Sachinidis, Agapios; Hescheler, Jürgen; Khalil, Markus; Halbach, Marcel

    2015-01-01

    Modelling of cardiac development, physiology and pharmacology by differentiation of embryonic stem cells (ESCs) requires comparability of cardiac differentiation between different ESC lines. To investigate whether the outcome of cardiac differentiation is consistent between different ESC lines, we compared electrophysiological properties of ESC-derived cardiomyocytes (ESC-CMs) of different murine ESC lines. Two wild-type (D3 and R1) and two transgenic ESC lines (D3/aPIG44 and CGR8/AMPIGX-7) were differentiated under identical culture conditions. The transgenic cell lines expressed enhanced green fluorescent protein (eGFP) and puromycin-N-acetyltransferase under control of the cardiac specific α-myosin heavy chain (αMHC) promoter. Action potentials (APs) were recorded using sharp electrodes and multielectrode arrays in beating clusters of ESC-CMs. Spontaneous AP frequency and AP duration (APD) as well as maximal upstroke velocity differed markedly between unpurified CMs of the four ESC lines. APD heterogeneity was negligible in D3/aPIG44, moderate in D3 and R1 and extensive in CGR8/AMPIGX-7. Interspike intervals calculated from long-term recordings showed a high degree of variability within and between recordings in CGR8/AMPIGX-7, but not in D3/aPIG44. Purification of the αMHC+ population by puromycin treatment posed only minor changes to APD in D3/aPIG44, but significantly shortened APD in CGR8/AMPIGX-7. Electrophysiological properties of ESC-CMs are strongly cell line-dependent and can be influenced by purification of cardiomyocytes by antibiotic selection. Thus, conclusions on cardiac development, physiology and pharmacology derived from single stem cell lines have to be interpreted carefully. © 2015 S. Karger AG, Basel.

  16. Establishment, Immortalisation and Characterisation of Pteropid Bat Cell Lines

    PubMed Central

    Crameri, Gary; Todd, Shawn; Grimley, Samantha; McEachern, Jennifer A.; Marsh, Glenn A.; Smith, Craig; Tachedjian, Mary; De Jong, Carol; Virtue, Elena R.; Yu, Meng; Bulach, Dieter; Liu, Jun-Ping; Michalski, Wojtek P.; Middleton, Deborah; Field, Hume E.; Wang, Lin-Fa

    2009-01-01

    Background Bats are the suspected natural reservoir hosts for a number of new and emerging zoonotic viruses including Nipah virus, Hendra virus, severe acute respiratory syndrome coronavirus and Ebola virus. Since the discovery of SARS-like coronaviruses in Chinese horseshoe bats, attempts to isolate a SL-CoV from bats have failed and attempts to isolate other bat-borne viruses in various mammalian cell lines have been similarly unsuccessful. New stable bat cell lines are needed to help with these investigations and as tools to assist in the study of bat immunology and virus-host interactions. Methodology/Findings Black flying foxes (Pteropus alecto) were captured from the wild and transported live to the laboratory for primary cell culture preparation using a variety of different methods and culture media. Primary cells were successfully cultured from 20 different organs. Cell immortalisation can occur spontaneously, however we used a retroviral system to immortalise cells via the transfer and stable production of the Simian virus 40 Large T antigen and the human telomerase reverse transcriptase protein. Initial infection experiments with both cloned and uncloned cell lines using Hendra and Nipah viruses demonstrated varying degrees of infection efficiency between the different cell lines, although it was possible to infect cells in all tissue types. Conclusions/Significance The approaches developed and optimised in this study should be applicable to bats of other species. We are in the process of generating further cell lines from a number of different bat species using the methodology established in this study. PMID:20011515

  17. Seven diverse human embryonic stem cell-derived chondrogenic clonal embryonic progenitor cell lines display site-specific cell fates.

    PubMed

    Sternberg, Hal; Kidd, Jennifer; Murai, James T; Jiang, Jianjie; Rinon, Ariel; Erickson, Isaac E; Funk, Walter D; Wang, Qian; Chapman, Karen B; Vangsness, C Thomas; West, Michael D

    2013-03-01

    The transcriptomes of seven diverse clonal human embryonic progenitor cell lines with chondrogenic potential were compared with that of bone marrow-derived mesenchymal stem cells (MSCs). The cell lines 4D20.8, 7PEND24, 7SMOO32, E15, MEL2, SK11 and SM30 were compared with MSCs using immunohistochemical methods, gene expression microarrays and quantitative real-time PCR. In the undifferentiated progenitor state, each line displayed unique combinations of site-specific markers, including AJAP1, ALDH1A2, BMP5, BARX1, HAND2, HOXB2, LHX1, LHX8, PITX1, TBX15 and ZIC2, but none of the lines expressed the MSC marker CD74. The lines showed diverse responses when differentiated in the presence of combinations of TGF-β3, BMP2, 4, 6 and 7 and GDF5, with the lines 4D20.8, SK11, SM30 and MEL2 showing osteogenic markers in some differentiation conditions. The line 7PEND24 showed evidence of regenerating articular cartilage and, in some conditions, markers of tendon differentiation. The scalability of site-specific clonal human embryonic stem cell-derived embryonic progenitor cell lines may provide novel models for the study of differentiation and methods for preparing purified and identified cells types for use in therapy.

  18. Human cell lines for biopharmaceutical manufacturing: history, status, and future perspectives

    PubMed Central

    Dumont, Jennifer; Euwart, Don; Mei, Baisong; Estes, Scott; Kshirsagar, Rashmi

    2016-01-01

    Abstract Biotherapeutic proteins represent a mainstay of treatment for a multitude of conditions, for example, autoimmune disorders, hematologic disorders, hormonal dysregulation, cancers, infectious diseases and genetic disorders. The technologies behind their production have changed substantially since biotherapeutic proteins were first approved in the 1980s. Although most biotherapeutic proteins developed to date have been produced using the mammalian Chinese hamster ovary and murine myeloma (NS0, Sp2/0) cell lines, there has been a recent shift toward the use of human cell lines. One of the most important advantages of using human cell lines for protein production is the greater likelihood that the resulting recombinant protein will bear post-translational modifications (PTMs) that are consistent with those seen on endogenous human proteins. Although other mammalian cell lines can produce PTMs similar to human cells, they also produce non-human PTMs, such as galactose-α1,3-galactose and N-glycolylneuraminic acid, which are potentially immunogenic. In addition, human cell lines are grown easily in a serum-free suspension culture, reproduce rapidly and have efficient protein production. A possible disadvantage of using human cell lines is the potential for human-specific viral contamination, although this risk can be mitigated with multiple viral inactivation or clearance steps. In addition, while human cell lines are currently widely used for biopharmaceutical research, vaccine production and production of some licensed protein therapeutics, there is a relative paucity of clinical experience with human cell lines because they have only recently begun to be used for the manufacture of proteins (compared with other types of cell lines). With additional research investment, human cell lines may be further optimized for routine commercial production of a broader range of biotherapeutic proteins. PMID:26383226

  19. Rapid micropatterning of cell lines and human pluripotent stem cells on elastomeric membranes.

    PubMed

    Paik, Isha; Scurr, David J; Morris, Bryan; Hall, Graham; Denning, Chris; Alexander, Morgan R; Shakesheff, Kevin M; Dixon, James E

    2012-10-01

    Tissue function during development and in regenerative medicine completely relies on correct cell organization and patterning at micro and macro scales. We describe a rapid method for patterning mammalian cells including human embryonic stem cells (HESCs) and induced pluripotent stem cells (iPSCs) on elastomeric membranes such that micron-scale control of cell position can be achieved over centimeter-length scales. Our method employs surface engineering of hydrophobic polydimethylsiloxane (PDMS) membranes by plasma polymerization of allylamine. Deposition of plasma polymerized allylamine (ppAAm) using our methods may be spatially restricted using a micro-stencil leaving faithful hydrophilic ppAAm patterns. We employed airbrushing to create aerosols which deposit extracellular matrix (ECM) proteins (such as fibronectin and Matrigel™) onto the same patterned ppAAm rich regions. Cell patterns were created with a variety of well characterized cell lines (e.g., NIH-3T3, C2C12, HL1, BJ6, HESC line HUES7, and HiPSC line IPS2). Individual and multiple cell line patterning were also achieved. Patterning remains faithful for several days and cells are viable and proliferate. To demonstrate the utility of our technique we have patterned cells in a variety of configurations. The ability to rapidly pattern cells at high resolution over macro scales should aid future tissue engineering efforts for regenerative medicine applications and in creating in vitro stem cell niches. Copyright © 2012 Wiley Periodicals, Inc.

  20. Incorrect strain information for mouse cell lines: sequential influence of misidentification on sublines.

    PubMed

    Uchio-Yamada, Kozue; Kasai, Fumio; Ozawa, Midori; Kohara, Arihiro

    2017-03-01

    Misidentification or cross-contamination of cell lines can cause serious issues. Human cell lines have been authenticated by short tandem repeat profiling; however, mouse cell lines have not been adequately assessed. In this study, mouse cell lines registered with the JCRB cell bank were examined by simple sequence length polymorphism (SSLP) analysis to identify their strains. Based on comparisons with 7 major inbred strains, our results revealed their strains in 80 of 90 cell lines. However, 12 of the 80 cell lines (15%) were found to differ from registered information. Of them, 4 cell lines originated from the same mouse, which had been generated through mating between two different inbred strains. The genotype of the mouse sample had not been examined after the backcross, leading to strain misidentification in those cell lines. Although 8 other cell lines had been established as sublines of a BALB/c cell line, their SSLP profiles are similar to a Swiss cell line. This affects differences in genotypes between inbred and outbred strains. Because the use of inbred samples and interbreeding between strains are not involved in human materials, our results suggest that the cause and influence of misidentification in mouse cell lines are different from those in human.

  1. HIV integration sites in latently infected cell lines: evidence of ongoing replication.

    PubMed

    Symons, Jori; Chopra, Abha; Malatinkova, Eva; De Spiegelaere, Ward; Leary, Shay; Cooper, Don; Abana, Chike O; Rhodes, Ajantha; Rezaei, Simin D; Vandekerckhove, Linos; Mallal, Simon; Lewin, Sharon R; Cameron, Paul U

    2017-01-13

    Assessing the location and frequency of HIV integration sites in latently infected cells can potentially inform our understanding of how HIV persists during combination antiretroviral therapy. We developed a novel high throughput sequencing method to evaluate HIV integration sites in latently infected cell lines to determine whether there was virus replication or clonal expansion in these cell lines observed as multiple integration events at the same position. We modified a previously reported method using random DNA shearing and PCR to allow for high throughput robotic processing to identify the site and frequency of HIV integration in latently infected cell lines. Latently infected cell lines infected with intact virus demonstrated multiple distinct HIV integration sites (28 different sites in U1, 110 in ACH-2 and 117 in J1.1 per 150,000 cells). In contrast, cell lines infected with replication-incompetent viruses (J-Lat cells) demonstrated single integration sites. Following in vitro passaging of the ACH-2 cell line, we observed a significant increase in the frequency of unique HIV integration sites and there were multiple mutations and large deletions in the proviral DNA. When the ACH-2 cell line was cultured with the integrase inhibitor raltegravir, there was a significant decrease in the number of unique HIV integration sites and a transient increase in the frequency of 2-LTR circles consistent with virus replication in these cells. Cell lines latently infected with intact HIV demonstrated multiple unique HIV integration sites indicating that these cell lines are not clonal and in the ACH-2 cell line there was evidence of low level virus replication. These findings have implications for the use of latently infected cell lines as models of HIV latency and for the use of these cells as standards.

  2. Efficacy of attractive toxic sugar baits (ATSB) against Aedes albopictus with garlic oil encapsulated in beta-cyclodextrin as the active ingredient

    PubMed Central

    Junnila, Amy; Revay, Edita E.; Müller, Gunter C.; Kravchenko, Vasiliy; Qualls, Whitney A.; Xue, Rui-de; Allen, Sandra A.; Beier, John C.; Schlein, Yosef

    2016-01-01

    We tested the efficacy of attractive toxic sugar bait (ATSB) with garlic oil microencapsulated in beta-cyclodextrin as active ingredient against Aedes albopictus in suburban Haifa, Israel. Two three-acre gardens with high numbers of Ae. albopictus were selected for perimeter spray treatment with ATSB and ASB (bait containing no active ingredient). Baits were colored with food dye to verify feeding of the mosquitoes. The mosquito population was monitored by human landing catches and sweep net catches in the surrounding vegetation. Experiments lasted for 44 days. Treatment occurred on day 13. The mosquito population collapsed about 4 days after treatment and continued to drop steadily for 27 days until the end of the study. At the experimental site the average pre-treatment landing rate was 17.2 per 5 mins. Two days post-treatment, the landing rate dropped to 11.4, and continued to drop to an average of 2.6 during the following 26 days. During the same period, the control population was stable. Few sugar fed females (8–10%) approached a human bait and anthrone tests showed relatively small amounts of sugar within their crop/gut. Around 60–70 % of males caught near our human bait were sugar positive which may indicate that the males were feeding on sugar for mating related behavior. From the vegetation treated with the toxic bait, we recovered significantly fewer (about 10–14%) males and females stained by ATSB than at the ASB-treated control. This may indicate that the toxic baits alter the resting behavior of the poisoned mosquitoes within the vegetation. Almost no Ae. albopictus females (5.2 ± 1.4) approached human bait after treatment with ATSB. It therefore appears that microencapsulated garlic oil is an effective pesticide against Ae. albopictus when used in an ATSB system. PMID:26403337

  3. Utility of human embryonic kidney cell line HEK-293 for rapid isolation of fixed and street rabies viruses: comparison with Neuro-2a and BHK-21 cell lines.

    PubMed

    Madhusudana, Shampur Narayan; Sundaramoorthy, Subha; Ullas, Padinjaremattatthil Thankappan

    2010-12-01

    A confirmatory rabies diagnosis can be achieved by rapid virus isolation in cell culture using brain tissue from the suspect animal. Several cell lines have been used for this purpose and the murine neuroblastoma cell line Neuro-2a has been found to be the most sensitive. The human embryonic kidney cell line HEK-293 is known to express several neuronal proteins and is believed to be of neuronal origin. We hypothesized that this cell line could be susceptible to rabies virus, which is highly neurotropic. First we tested the sensitivity of HEK-293 cells to the laboratory strain, challenge virus standard (CVS). We then tested 120 brain samples from different animals and humans suspected to have died of rabies by fluorescent antibody test (FAT). Both FAT-positive and FAT-negative brains were tested for virus isolation using Neuro-2a, BHK-21, and HEK-293 cell lines and also by mouse inoculation. There was 100% correlation between FAT, virus isolation in Neuro-2a and HEK-293 cells, and mouse inoculation. However, the rate of virus isolation in the BHK-21 cell line was only 28% when compared to the other cell lines. The sensitivity of HEK-293 to CVS strain of virus was similar to that of Neuro-2a. We conclude that the HEK-293 cell line is as sensitive as the Neuro-2a cell line for the rapid isolation of rabies virus and may serve as an alternative cell line for rabies diagnosis and future research. Copyright © 2010 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  4. Characteristics of bovine inner cell mass-derived cell lines and their fate in chimeric conceptuses.

    PubMed

    Furusawa, Tadashi; Ohkoshi, Katsuhiro; Kimura, Koji; Matsuyama, Shuichi; Akagi, Satoshi; Kaneda, Masahiro; Ikeda, Mitsumi; Hosoe, Misa; Kizaki, Keiichiro; Tokunaga, Tomoyuki

    2013-08-01

    Bovine embryonic stem (ES) cells have the potential to provide significant benefits in a range of agricultural and biomedical applications. Here, we employed a combination of conventional methods using glycogen synthase kinase 3 and mitogen-activated protein kinase inhibitors to establish ES cell lines from in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) bovine embryos. Five male cell lines were established from IVF embryos, and two female and three male cell lines from SCNT blastocysts; we named these lines bovine ES cell-like cells (bESLCs). The lines exhibited dome-shaped colonies, stained positively for alkaline phosphatase, and expressed pluripotent stem cell markers such as POU5F1, SOX2, and SSEA-1. The expression levels of these markers, especially for NANOG, varied among the cell lines. A DNA methylation assay showed the POU5F1 promoter region was hypomethylated compared to fibroblast cells. An in vitro differentiation assay showed that endoderm and ectoderm marker genes, but not mesoderm markers, were upregulated in differentiating bESLCs. To examine bESLCs in later embryonic stages, we created 22 chimeric blastocysts with a male bESLC line carrying a GFP marker gene and transferred these to a recipient cow. Four chimeric embryos were subsequently retrieved on Day 13 and retransferred to two recipient cows. One living fetus was obtained at Day 62. GFP signals were not identified in fetal cells by fluorescence microscopy; however, genomic PCR analysis detected the GFP gene in major organs. Clusters of GFP-positive cells were observed in amniotic membranes, suggesting that bESLCs can be categorized as a novel type of ICM-derived cells that can potentially differentiate into epiblast and hypoblast lineages.

  5. Effect of Common Species of Florida Landscaping Plants on the Efficacy of Attractive Toxic Sugar Baits Against Aedes albopictus.

    PubMed

    Seeger, Kelly E; Scott, Jodi M; Muller, Gunter C; Qualls, Whitney A; Xue, Rui-De

    2017-06-01

    Attractive toxic sugar bait (ATSB) was applied to 5 different types of commonly found plants in landscaping of northeastern Florida. The ATSB applications were assessed for possible plant effects and preference against Aedes albopictus in semifield evaluations. Positive and negative controls consisted of plants sprayed with attractive sugar bait (no toxicant) and plants with nothing applied. Bioassays were conducted on stems with leaf clippings and on full plants to assess any difference in mosquito mortality on the different plants. Plants utilized in these evaluations were Indian hawthorne, Yaupon holly, Japanese privet, Loropetalum ruby, and podocarpus. In both assays, no significant difference was observed in the effect of ATSBs on adult female mosquitoes based on the type of plant. ATSB could be applied to common landscape plants for adult Ae. albopictus control.

  6. Establishment and characterization of a novel lymphangiosarcoma cell line (MO-LAS) compared with the hemangiosarcoma cell line (ISO-HAS).

    PubMed

    Masuzawa, Mikio; Masuzawa, Mamiko; Hamada, Yuhko; Arakawa, Nobuko; Mori, Mari; Ishii, Masako; Nishiyama, Shigeo

    2012-08-01

    The concept of "lymphangiosarcoma" remains obscure. Therefore, we reported a patient with lymphangiosarcoma, resistant to immunotherapy. The patient presented with impressive and discriminative features: clinically an ill-defined edematous lesion with lymphorrhea and pathologically atypical vascular channel formation without extravasation of blood, clearly distinguished from common angiosarcoma with hemorrhage. From this case, a lymphangiosarcoma cell line, MO-LAS, was established and its characteristics were compared with the hemangiosarcoma cell line, ISO-HAS. Flow cytometric analysis revealed that MO-LAS was negative for factor VIII-related antigen, but positive for CD31, D2-40, NZ-1, and vascular endothelial growth factor receptor-3 (VEGFR-3), similar to ISO-HAS. However, MO-LAS expressed a much higher level of homeobox gene PROX1, indicating a lymphatic phenotype, compared with ISO-HAS. Furthermore, MO-LAS showed a much lesser expression of oncogenes and much lower sensitivity against lymphokine-activated killer (LAK) cells. Lymphangiosarcoma may be difficult to recognize by the immune system. Conclusively, the establishment of MO-LAS, a novel angiosarcoma cell line bearing lymphatic characters, strongly suggests the entity of lymphangiosarcoma.

  7. SENSORY HAIR CELL REGENERATION IN THE ZEBRAFISH LATERAL LINE

    PubMed Central

    Lush, Mark E.; Piotrowski, Tatjana

    2014-01-01

    Damage or destruction of sensory hair cells in the inner ear leads to hearing or balance deficits that can be debilitating, especially in older adults. Unfortunately, the damage is permanent, as regeneration of the inner ear sensory epithelia does not occur in mammals. Zebrafish and other non-mammalian vertebrates have the remarkable ability to regenerate sensory hair cells and understanding the molecular and cellular basis for this regenerative ability will hopefully aid us in designing therapies to induce regeneration in mammals. Zebrafish not only possess hair cells in the ear but also in the sensory lateral line system. Hair cells in both organs are functionally analogous to hair cells in the inner ear of mammals. The lateral line is a mechanosensory system found in most aquatic vertebrates that detects water motion and aids in predator avoidance, prey capture, schooling and mating. Although hair cell regeneration occurs in both the ear and lateral line, most research to date has focused on the lateral line due to its relatively simple structure and accessibility. Here we review the recent discoveries made during the characterization of hair cell regeneration in zebrafish. PMID:25045019

  8. Sensory hair cell regeneration in the zebrafish lateral line.

    PubMed

    Lush, Mark E; Piotrowski, Tatjana

    2014-10-01

    Damage or destruction of sensory hair cells in the inner ear leads to hearing or balance deficits that can be debilitating, especially in older adults. Unfortunately, the damage is permanent, as regeneration of the inner ear sensory epithelia does not occur in mammals. Zebrafish and other non-mammalian vertebrates have the remarkable ability to regenerate sensory hair cells and understanding the molecular and cellular basis for this regenerative ability will hopefully aid us in designing therapies to induce regeneration in mammals. Zebrafish not only possess hair cells in the ear but also in the sensory lateral line system. Hair cells in both organs are functionally analogous to hair cells in the inner ear of mammals. The lateral line is a mechanosensory system found in most aquatic vertebrates that detects water motion and aids in predator avoidance, prey capture, schooling, and mating. Although hair cell regeneration occurs in both the ear and lateral line, most research to date has focused on the lateral line due to its relatively simple structure and accessibility. Here we review the recent discoveries made during the characterization of hair cell regeneration in zebrafish. Copyright © 2014 Wiley Periodicals, Inc.

  9. Integrated analysis of miRNAs and transcriptomes in Aedes albopictus midgut reveals the differential expression profiles of immune-related genes during dengue virus serotype-2 infection.

    PubMed

    Liu, Yan-Xia; Li, Fen-Xiang; Liu, Zhuan-Zhuan; Jia, Zhi-Rong; Zhou, Yan-He; Zhang, Hao; Yan, Hui; Zhou, Xian-Qiang; Chen, Xiao-Guang

    2016-06-01

    Mosquito microRNAs (miRNAs) are involved in host-virus interaction, and have been reported to be altered by dengue virus (DENV) infection in Aedes albopictus (Diptera: Culicidae). However, little is known about the molecular mechanisms of Aedes albopictus midgut-the first organ to interact with DENV-involved in its resistance to DENV. Here we used high-throughput sequencing to characterize miRNA and messenger RNA (mRNA) expression patterns in Aedes albopictus midgut in response to dengue virus serotype 2. A total of three miRNAs and 777 mRNAs were identified to be differentially expressed upon DENV infection. For the mRNAs, we identified 198 immune-related genes and 31 of them were differentially expressed. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses also showed that the differentially expressed immune-related genes were involved in immune response. Then the differential expression patterns of six immune-related genes and three miRNAs were confirmed by real-time reverse transcription polymerase chain reaction. Furthermore, seven known miRNA-mRNA interaction pairs were identified by aligning our two datasets. These analyses of miRNA and mRNA transcriptomes provide valuable information for uncovering the DENV response genes and provide a basis for future study of the resistance mechanisms in Aedes albopictus midgut. © 2016 Institute of Zoology, Chinese Academy of Sciences.

  10. Human renal cell carcinoma: establishment and characterization of two new cell lines.

    PubMed

    Naito, S; Kanamori, T; Hisano, S; Tanaka, K; Momose, S; Kamata, N

    1982-11-01

    Characterization studies have been carried out on 2 cell lines (KPK 1 and KPK 13) established from human renal adenocarcinoma. KPK 1 and KPK 13 have been passaged 178 times in vitro for about 6 years and 7 months and 78 times for about 3 years an 2 months, respectively. Although morphologic differences exist between the 2 lines, each has an epithelial morphology and exhibits multilayering. Doubling time of KPK 1 and KPK 13 cells was 29 hours and 51 hours, respectively. Both KPK 1 and KPK 13 induced tumors at the site of subcutaneous injection, closely resembling the original tumor from which they were derived. Chromosome number of both cell lines was 100 per cent aneuploid and the presence of Y chromosomes was confirmed by G banding in KPK 13 cells. KPK 1 was found to have high thromboplastic and high fibrinolytic activities, whereas KPK 13 was shown to have comparatively low thromboplastic and no detectable fibrinolytic activities. These activities were detected in the serum free supernatant fraction from KPK 1 cells but were not detected in that from KPK 13 cells.

  11. Tumourigenic canine osteosarcoma cell lines associated with frizzled-6 up-regulation and enhanced side population cell frequency.

    PubMed

    de Sá Rodrigues, L C; Holmes, K E; Thompson, V; Newton, M A; Stein, T J

    2017-03-01

    An increased serum alkaline phosphatase concentration is known to be associated with a negative prognosis in canine and human osteosarcoma. To expand upon previous studies regarding the biological relevance of increased serum alkaline phosphatase as a negative prognostic factor, xenogeneic heterotopic transplants were performed using six canine primary osteosarcoma cell lines generated from patients with differing serum alkaline phosphatase concentrations (three normal and three increased). Three of the six cell lines were capable of generating tumours and tumour formation was independent of the serum alkaline phosphatase status of the cell line. Microarray analysis identified 379 genes as being differentially expressed between the tumourigenic and non-tumourigenic cell lines. Frizzled-6 was upregulated to the greatest extent (7.78-fold) in tumourigenic cell lines compared with non-tumourigenic cell lines. Frizzled-6, a co-receptor for Wnt ligands has been associated with enhanced tumour-initiating cells and poor prognosis for other tumours. The increased expression of frizzled-6 was confirmed by quantitative reverse transcription polymerase chain reaction (QPCR) and Western blot analysis. Additionally, the tumourigenic cell lines also had an increase in the percentage of side population cells compared with non-tumourigenic cell lines (5.89% versus 1.58%, respectively). There were no differences in tumourigenicity, frizzled-6 or percentage of side population cells noted between osteosarcoma cell lines generated from patients of differing serum alkaline phosphatase concentration. However, to our knowledge this is the first study to identified frizzled-6 as a possible marker of osteosarcoma cell populations with enhanced tumourigenicity and side population cells. Future work will focus on defining the role of frizzled-6 in osteosarcoma tumourigenesis and tumour-initiating cells. © 2015 John Wiley & Sons Ltd.

  12. Host-seeking height preferences of Aedes albopictus (Diptera: Culicidae) in North-central Florida suburban and sylvatic locales

    USDA-ARS?s Scientific Manuscript database

    The response of Aedes albopictus to the BG-Sentinel™, Omni-directional-Fay-Prince and Mosquito Magnet-X traps was evaluated in four suburban and four sylvatic habitats in North-Central Florida to ascertain potential height preference of this species. These traps, which are primarily designed to att...

  13. Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients.

    PubMed

    Ostrow, Kimberly Laskie; Donaldson, Katelyn; Blakeley, Jaishri; Belzberg, Allan; Hoke, Ahmet

    2015-01-01

    Schwannomatosis, a rare form of neurofibromatosis, is characterized predominantly by multiple, often painful, schwannomas throughout the peripheral nervous system. The current standard of care for schwannomatosis is surgical resection. A major obstacle to schwannomatosis research is the lack of robust tumor cell lines. There is a great need for mechanistic and drug discovery studies of schwannomatosis, yet appropriate tools are not currently available. Schwannomatosis tumors are difficult to grow in culture as they survive only a few passages before senescence. Our lab has extensive experience in establishing primary and immortalized human Schwann cell cultures from normal tissue that retain their phenotypes after immortalization. Therefore we took on the challenge of creating immortalized human Schwann cell lines derived from tumors from schwannomatosis patients. We have established and fully characterized 2 schwannomatosis cell lines from 2 separate patients using SV40 virus large T antigen. One patient reported pain and the other did not. The schwannomatosis cell lines were stained with S100B antibodies to confirm Schwann cell identity. The schwannomatosis cells also expressed the Schwann cell markers, p75NTR, S100B, and NGF after multiple passages. Cell morphology was retained following multiple passaging and freeze/ thaw cycles. Gene expression microarray analysis was used to compare the cell lines with their respective parent tumors. No differences in key genes were detected, with the exception that several cell cycle regulators were upregulated in the schwannomatosis cell lines when compared to their parent tumors. This upregulation was apparently a product of cell culturing, as the schwannomatosis cells exhibited the same expression pattern of cell cycle regulatory genes as normal primary human Schwann cells. Cell growth was also similar between normal primary and immortalized tumor cells in culture. Accurate cell lines derived directly from human tumors

  14. Immortalized Human Schwann Cell Lines Derived From Tumors of Schwannomatosis Patients

    PubMed Central

    Ostrow, Kimberly Laskie; Donaldson, Katelyn; Blakeley, Jaishri; Belzberg, Allan; Hoke, Ahmet

    2015-01-01

    Schwannomatosis, a rare form of neurofibromatosis, is characterized predominantly by multiple, often painful, schwannomas throughout the peripheral nervous system. The current standard of care for schwannomatosis is surgical resection. A major obstacle to schwannomatosis research is the lack of robust tumor cell lines. There is a great need for mechanistic and drug discovery studies of schwannomatosis, yet appropriate tools are not currently available. Schwannomatosis tumors are difficult to grow in culture as they survive only a few passages before senescence. Our lab has extensive experience in establishing primary and immortalized human Schwann cell cultures from normal tissue that retain their phenotypes after immortalization. Therefore we took on the challenge of creating immortalized human Schwann cell lines derived from tumors from schwannomatosis patients. We have established and fully characterized 2 schwannomatosis cell lines from 2 separate patients using SV40 virus large T antigen. One patient reported pain and the other did not. The schwannomatosis cell lines were stained with S100B antibodies to confirm Schwann cell identity. The schwannomatosis cells also expressed the Schwann cell markers, p75NTR, S100B, and NGF after multiple passages. Cell morphology was retained following multiple passaging and freeze/ thaw cycles. Gene expression microarray analysis was used to compare the cell lines with their respective parent tumors. No differences in key genes were detected, with the exception that several cell cycle regulators were upregulated in the schwannomatosis cell lines when compared to their parent tumors. This upregulation was apparently a product of cell culturing, as the schwannomatosis cells exhibited the same expression pattern of cell cycle regulatory genes as normal primary human Schwann cells. Cell growth was also similar between normal primary and immortalized tumor cells in culture. Accurate cell lines derived directly from human tumors

  15. Generation and Characterization of JCV Permissive Hybrid Cell Lines

    PubMed Central

    Sariyer, Ilker K.; Safak, Mahmut; Gordon, Jennifer; Khalili, Kamel

    2009-01-01

    JC virus (JCV) is a human neurotropic polyomavirus whose replication in the central nervous system induces the fatal demyelinating disease, progressive multifocal leukoencephalopathy (PML). JCV particles have been detected primarily in oligodendrocytes and astrocytes of the brains of patients with PML and in the laboratory its propagation is limited to primary cultures of human fetal glial cells. In this short communication, the development of a new cell culture system is described through the fusion of primary human fetal astrocytes with the human glioblastoma cell line, U-87MG. The new hybrid cell line obtained from this fusion has the capacity to support efficiently expression of JCV and replication of viral DNA in vitro up to 16 passages. This cell line can serve as a reliable culture system to study the biology of JCV host cell interaction, determine the mechanisms involved in cell type specific replication of JCV, and provide a convenient cell culture system for high throughput screening of anti-viral agents. PMID:19442856

  16. Astaxanthin Inhibits Proliferation of Human Gastric Cancer Cell Lines by Interrupting Cell Cycle Progression

    PubMed Central

    Kim, Jung Ha; Park, Jong-Jae; Lee, Beom Jae; Joo, Moon Kyung; Chun, Hoon Jai; Lee, Sang Woo; Bak, Young-Tae

    2016-01-01

    Background/Aims Astaxanthin is a carotenoid pigment that has antioxidant, antitumoral, and anti-inflammatory properties. In this in vitro study, we investigated the mechanism of anticancer effects of astaxanthin in gastric carcinoma cell lines. Methods The human gastric adenocarcinoma cell lines AGS, KATO-III, MKN-45, and SNU-1 were treated with various concentrations of astaxanthin. A cell viability test, cell cycle analysis, and immunoblotting were performed. Results The viability of each cancer cell line was suppressed by astaxanthin in a dose-dependent manner with significantly decreased proliferation in KATO-III and SNU-1 cells. Astaxanthin increased the number of cells in the G0/G1 phase but reduced the proportion of S phase KATO-III and SNU-1 cells. Phosphorylated extracellular signal-regulated kinase (ERK) was decreased in an inverse dose-dependent correlation with astaxanthin concentration, and the expression of p27kip-1 increased the KATO-III and SNU-1 cell lines in an astaxanthin dose-dependent manner. Conclusions Astaxanthin inhibits proliferation by interrupting cell cycle progression in KATO-III and SNU-1 gastric cancer cells. This may be caused by the inhibition of the phosphorylation of ERK and the enhanced expression of p27kip-1. PMID:26470770

  17. Recombinant protein production from stable mammalian cell lines and pools.

    PubMed

    Hacker, David L; Balasubramanian, Sowmya

    2016-06-01

    We highlight recent developments for the production of recombinant proteins from suspension-adapted mammalian cell lines. We discuss the generation of stable cell lines using transposons and lentivirus vectors (non-targeted transgene integration) and site-specific recombinases (targeted transgene integration). Each of these methods results in the generation of cell lines with protein yields that are generally superior to those achievable through classical plasmid transfection that depends on the integration of the transfected DNA by non-homologous DNA end-joining. This is the main reason why these techniques can also be used for the generation of stable cell pools, heterogenous populations of recombinant cells generated by gene delivery and genetic selection without resorting to single cell cloning. This allows the time line from gene transfer to protein production to be reduced. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Trichloroethylene toxicity in a human hepatoma cell line

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Thevenin, E.; McMillian, J.

    1994-12-31

    The experiments conducted in this study were designed to determine the usefullness of hepatocyte cultures and a human hepatoma cell line as model systems for assessing human susceptibility to hepatocellular carcinoma due to exposure to trichloroethylene. The results from these studies will then be analyzed to determine if human cell lines can be used to conduct future experiments of this nature.

  19. Reference Maps of Human ES and iPS Cell Variation Enable High-Throughput Characterization of Pluripotent Cell Lines

    PubMed Central

    Bock, Christoph; Kiskinis, Evangelos; Verstappen, Griet; Gu, Hongcang; Boulting, Gabriella; Smith, Zachary D.; Ziller, Michael; Croft, Gist F.; Amoroso, Mackenzie W.; Oakley, Derek H.; Gnirke, Andreas; Eggan, Kevin; Meissner, Alexander

    2011-01-01

    SUMMARY The developmental potential of human pluripotent stem cells suggests that they can produce disease-relevant cell types for biomedical research. However, substantial variation has been reported among pluripotent cell lines, which could affect their utility and clinical safety. Such cell-line-specific differences must be better understood before one can confidently use embryonic stem (ES) or induced pluripotent stem (iPS) cells in translational research. Toward this goal we have established genome-wide reference maps of DNA methylation and gene expression for 20 previously derived human ES lines and 12 human iPS cell lines, and we have measured the in vitro differentiation propensity of these cell lines. This resource enabled us to assess the epigenetic and transcriptional similarity of ES and iPS cells and to predict the differentiation efficiency of individual cell lines. The combination of assays yields a scorecard for quick and comprehensive characterization of pluripotent cell lines. PMID:21295703

  20. Assessment of Cell Line Models of Primary Human Cells by Raman Spectral Phenotyping

    PubMed Central

    Swain, Robin J.; Kemp, Sarah J.; Goldstraw, Peter; Tetley, Teresa D.; Stevens, Molly M.

    2010-01-01

    Abstract Researchers have previously questioned the suitability of cell lines as models for primary cells. In this study, we used Raman microspectroscopy to characterize live A549 cells from a unique molecular biochemical perspective to shed light on their suitability as a model for primary human pulmonary alveolar type II (ATII) cells. We also investigated a recently developed transduced type I (TT1) cell line as a model for alveolar type I (ATI) cells. Single-cell Raman spectra provide unique biomolecular fingerprints that can be used to characterize cellular phenotypes. A multivariate statistical analysis of Raman spectra indicated that the spectra of A549 and TT1 cells are characterized by significantly lower phospholipid content compared to ATII and ATI spectra because their cytoplasm contains fewer surfactant lamellar bodies. Furthermore, we found that A549 spectra are statistically more similar to ATI spectra than to ATII spectra. The spectral variation permitted phenotypic classification of cells based on Raman spectral signatures with >99% accuracy. These results suggest that A549 cells are not a good model for ATII cells, but TT1 cells do provide a reasonable model for ATI cells. The findings have far-reaching implications for the assessment of cell lines as suitable primary cellular models in live cultures. PMID:20409492

  1. Two clonal cell lines of immortalized human corneal endothelial cells show either differentiated or precursor cell characteristics.

    PubMed

    Valtink, Monika; Gruschwitz, Rita; Funk, Richard H W; Engelmann, Katrin

    2008-01-01

    Access to primary human corneal endothelial cells (HCEC) is limited and donor-derived differences between cultures exacerbate the issue of data reproducibility, whereas cell lines can provide sufficient numbers of homogenous cells for multiple experiments. An immortalized HCEC population was adapted to serum-free culture medium and repeated cloning was performed. Clonally grown cells were propagated under serum-free conditions and growth curves were recorded. Cells were characterized immunocytochemically for junctional proteins, collagens, Na,K-ATPase and HCEC-specific 9.3.E-antigen. Ultrastructure was monitored by scanning and transmission electron microscopy. Two clonal cell lines, HCEC-B4G12 and HCEC-H9C1, could be isolated and expanded, which differed morphologically: B4G12 cells were polygonal, strongly adherent and formed a strict monolayer, H9C1 cells were less adherent and formed floating spheres. The generation time of B4G12 cells was 62.26 +/- 14.5 h and that of H9C1 cells 44.05 +/- 5.05 h. Scanning electron microscopy revealed that B4G12 cells had a smooth cell surface, while H9C1 cells had numerous thin filopodia. Both cell lines expressed ZO-1 and occludin adequately, and little but well detectable amounts of connexin-43. Expression of HCEC-specific 9.3.E-antigen was found commensurately in both cell lines, while expression of Na,K-ATPase alpha1 was higher in H9C1 cells than in B4G12 cells. B4G12 cells expressed collagen IV abundantly and almost no collagen III, while H9C1 cells expressed both collagens at reasonable amounts. It is concluded that the clonal cell line B4G12 represents an ideal model of differentiated HCEC, while H9C1 may reflect features of developing or transitional HCEC. Copyright 2008 S. Karger AG, Basel.

  2. Establishment of stable cell line for inducing KAP1 protein expression.

    PubMed

    Liu, Xiaoyan; Khan, Md Asaduzzaman; Cheng, Jingliang; Wei, Chunli; Zhang, Lianmei; Fu, Junjiang

    2015-06-01

    Generation of the stable cell lines is a highly efficient tool in functional studies of certain genes or proteins, where the particular genes or proteins are inducibly expressed. The KRAB-associated protein-1 (KAP1) is an important transcription regulatory protein, which is investigated in several molecular biological studies. In this study, we have aimed to generate a stable cell line for inducing KAP1 expression. The recombinant plasmid pcDNA5/FRT/TO-KAP1 was constructed at first, which was then transfected into Flp-In™T-REx™-HEK293 cells to establish an inducible pcDNA5/FRT/TO-KAP1-HEK293 cell line. The Western blot analysis showed that the protein level of KAP1 is over-expressed in the established stable cell line by doxycycline induction, both dose and time dependently. Thus we have successfully established stable pcDNA5/FRT/TO-KAP1-HEK293 cell line, which can express KAP1 inducibly. This inducible cell line might be very useful for KAP1 functional studies.

  3. Reliable in vitro studies require appropriate ovarian cancer cell lines

    PubMed Central

    2014-01-01

    Ovarian cancer is the fifth most common cause of cancer death in women and the leading cause of death from gynaecological malignancies. Of the 75% women diagnosed with locally advanced or disseminated disease, only 30% will survive five years following treatment. This poor prognosis is due to the following reasons: limited understanding of the tumor origin, unclear initiating events and early developmental stages of ovarian cancer, lack of reliable ovarian cancer-specific biomarkers, and drug resistance in advanced cases. In the past, in vitro studies using cell line models have been an invaluable tool for basic, discovery-driven cancer research. However, numerous issues including misidentification and cross-contamination of cell lines have hindered research efforts. In this study we examined all ovarian cancer cell lines available from cell banks. Hereby, we identified inconsistencies in the reporting, difficulties in the identification of cell origin or clinical data of the donor patients, restricted ethnic and histological type representation, and a lack of tubal and peritoneal cancer cell lines. We recommend that all cell lines should be distributed via official cell banks only with strict guidelines regarding the minimal available information required to improve the quality of ovarian cancer research in future. PMID:24936210

  4. Impact of Tolypocladium cylindrosporum (Hypocreales: Ophiocordycipitaceae) on eggs of Aedes aegypti and Aedes albopictus at low temperature

    USDA-ARS?s Scientific Manuscript database

    We examined the potential of Tolypocladium cylindrosporum IBT 41712 to infect eggs of Aedes aegypti Linnaeus and Aedes albopictus (Skuse) mosquitoes at low temperature (15 deg C). To determine the optimum temperature for the fungus, we cultured the fungus at eight temperatures (4, 12, 15, 21, 28, 33...

  5. Outcomes from the USDA/ARS area-wide project for management of the Asian tiger mosquito, Aedes albopictus

    USDA-ARS?s Scientific Manuscript database

    The Asian tiger mosquito, Aedes albopictus, became established in the continental US in 1985 and now infests 30 states. In 2007 the USDA Agricultural Research Service funded an “area-wide” project focused on the management of this species. The project was a unique federal, state, local collaborati...

  6. Effect of application rate and persistence of boric acid sugar baits applied to plants for control of Aedes albopictus.

    PubMed

    Xue, Rui-De; Müller, Günter C; Kline, Daniel L; Barnard, Donald R

    2011-03-01

    The use of toxic bait to kill adult Aedes albopictus is a safe and potentially effective alternative to the use of synthetic chemical insecticides. This study was carried out to determine effective concentrations of boric acid needed in sugar bait solutions applied to plant surfaces, and to determine its residual effect in reducing adult mosquito densities. In outdoor tests in 1,100-m3 screened enclosures, landing rates of Ae. albopictus on a human subject and the number of female mosquitoes in mechanical traps were significantly reduced by a 1% boric acid bait compared with the other tested concentrations (0.25%, 0.50%, and 0.75%) and untreated control. Studies of the duration of boric acid activity on plant surfaces were made in 1.4-m3 cages in the laboratory and outdoors in 78-m3 screened enclosures. In the laboratory tests, 1% boric acid bait resulted in >96% mortality in male and female Ae. albopictus for 14 days, whereas in outdoor tests, mosquito landing rates in the treated enclosures were significantly lower than in the control enclosures for 7 days. Also, mosquito mortality responses to boric acid baits between plants with flowers and nonflowers (1.4-m3 cages in the laboratory) were not significantly different. The results of this study suggest that boric acid baits applied to plant surfaces may provide specific data related to the development of an effective point-source-based adjunct/alternative to the use of conventional adulticides for mosquito control.

  7. Effects of elevated atmospheric CO2 on competition between the mosquitoes Aedes albopictus and Ae. triseriatus via changes in litter quality and production.

    PubMed

    Smith, C; Baldwin, A H; Sullivan, J; Leisnham, P T

    2013-05-01

    Elevated atmospheric CO2 can alter aquatic communities via changes in allochthonous litter inputs. We tested effects of atmospheric CO2 on the invasive Aedes albopictus (Skuse) and native Aedes triseriatus (Say) (Diptera: Culicidae) via changes in competition for microbial food or resource inhibition/toxicity. Quercus alba L. litter was produced under elevated (879 ppm) and ambient (388 ppm) atmospheric CO2. Saplings grown at elevated CO2 produced greater litter biomass, which decayed faster and leached more tannins than saplings at ambient CO2. Competition was tested by raising larvae in different species and density combinations provisioned with elevated- or ambient-CO2 litter. Species-specific performance to water conditions was tested by providing single-species larval cohorts with increasing amounts of elevated- or ambient-CO2 litter, or increasing concentrations of tannic acid. Larval densities affected some fitness parameters of Ae. albopictus and Ae. triseriatus, but elevated-CO2 litter did not modify the effects of competition on population growth rates or any fitness parameters. Population growth rates and survival of each species generally were affected negatively by increasing amounts of both elevated- and ambient-CO2 litter from 0.252 to 2.016 g/liter, and tannic acid concentrations above 100 mg/liter were entirely lethal to both species. Aedes albopictus had consistently higher population growth rates than Ae. triseriatus. These results suggest that changes to litter production and chemistry from elevated CO2 are unlikely to affect the competitive outcome between Ae. albopictus and Ae. triseriatus, but that moderate increases in litter production increase population growth rates of both species until a threshold is exceeded that results in resource inhibition and toxicity.

  8. Escherichia coli STb toxin induces apoptosis in intestinal epithelial cell lines.

    PubMed

    Syed, H Claudia; Dubreuil, J Daniel

    2012-09-01

    A previous study conducted in our laboratory demonstrated that cells having internalized Escherichia coli STb toxin display apoptotic-like morphology. We therefore investigated if STb could induce programmed cell death in both a human and an animal intestinal epithelial cell lines. HRT-18 (Human Colon Tumor) and IEC-18 (Rat Ileum Epithelial Cells) cell lines were used. As STb is frequently tested in a rat model, the IEC-18 cell line was most relevant to our work. The cell lines were treated with various amounts of purified STb (nanomole range) for a period of 24 h after which cells were harvested and examined for apoptotic characteristics. Caspase-9, the initiator of mitochondrion-mediated apoptosis, and caspase-3, an effector of caspase-9, were both activated following STb intoxication of HRT-18 and IEC-18 cells whereas caspase-8, the initiator caspase of the extrinsic pathway, was not activated. For both cell lines, agarose gel electrophoresis of the cell DNA content reveals laddering of DNA, resulting from DNA fragmentation, a characteristic of apoptosis. Hoechst 33342-stained DNA of STb-treated cell lines, observed using fluorescence microscopy, revealed condensation and fragmentation of the nuclei. Apoptotic indexes calculated from fragmented nuclei of Hoechst 33342-stained DNA for HRT-18 and IEC-18 cells showed an STb dose-dependent response. Overall, these data indicate that STb toxin induces a mitochondrion-mediated caspase-dependent apoptotic pathway. Copyright © 2012 Elsevier Ltd. All rights reserved.

  9. Cytotoxic effects of treosulfan on prostate cancer cell lines.

    PubMed

    Feyerabend, Susan; Feil, Gerhard; Krug, Jutta; Kassen, Annette; Stenzl, Arnulf

    2007-01-01

    Despite various therapeutical options in metastatic prostate cancer, the lack of a curative approach motivates further investigations. Treosulfan is an alkylating agent that has proven its indication in the treatment of e.g. ovarian carcinoma. This study focused on the objective of evaluating the effect of in vitro intoxication of human prostate carcinoma cell lines with treosulfan. Human prostate cancer cell lines LNCaP, DU145 and PC3 were treated with treosulfan concentrations from 0.5-500 microM for up to six days. Analysis of cell viability was performed using colorimetric WST-1 assay. Control data were obtained from identical cell lines cultivated without treosulfan. Incubation with treosulfan inhibited cell viability and led to cell death in all cell lines in a dose- and time-dependent manner. After one day, viability of LNCaP, DU145 and PC3 cells was constantly reduced with a dose rate of at least 10 microM (p < 0.001), 10 microM (p < 0.0001) and 100 microM (p < 0.0001) treosulfan, respectively. Minimum dose rates leading to death of nearly all LNCaP, DU145 and PC3 cells were 250 microM, 100 microM and 200 microM treosulfan, respectively. The results demonstrate a sensitivity of prostate carcinoma cells to the cytotoxic activity of treosulfan. Therefore, treosulfan might be a promising compound for novel treatment protocols for prostate cancer.

  10. [Effects of ezrin silencing on pancreatic cancer cell line Panc-1].

    PubMed

    Meng, Yun-xiao; Yu, Shuang-ni; Lu, Zhao-hui; Chen, Jie

    2012-12-01

    To explore the effects of ezrin silencing on pancreatic cancer cell line Panc-1. Pancreatic cancer cell line Panc-1 was transfected with ezrin silencing plasmid. The proliferation and the cell cycle status were determined by CCK-8 assay and flow cytometry analysis, respectively. Cellular membrane protrusions/microvilli formation were visualized by scanning election microscopy. Colony formation assay was used to determine the cell anchor-independent growth ability in vitro. Trans-filter migration and invasion assays were performed with 8 µm pore inserts in a 24-well BioCoat chamber with/without Matrigel. Ezrin silencing decreased cellular protrusions/microvilli formation, anchorage-independent growth, cell migration and invasion, but had no effects on cell proliferation in vitro and cell cycle, in pancreatic cancer cell line Panc-1. Ezrin expression affects the cellular protrusions/microvilli formation, anchorage-independent growth, cell migration and invasion in pancreatic cancer cell line Panc-1.

  11. Derivation of Human Skin Fibroblast Lines for Feeder Cells of Human Embryonic Stem Cells.

    PubMed

    Unger, Christian; Felldin, Ulrika; Rodin, Sergey; Nordenskjöld, Agneta; Dilber, Sirac; Hovatta, Outi

    2016-02-03

    After the first derivations of human embryonic stem cell (hESC) lines on fetal mouse feeder cell layers, the idea of using human cells instead of mouse cells as feeder cells soon arose. Mouse cells bear a risk of microbial contamination, and nonhuman immunogenic proteins are absorbed from the feeders to hESCs. Human skin fibroblasts can be effectively used as feeder cells for hESCs. The same primary cell line, which can be safely used for up to 15 passages after stock preparations, can be expanded and used for large numbers of hESC derivations and cultures. These cells are relatively easy to handle and maintain. No animal facilities or animal work is needed. Here, we describe the derivation, culture, and cryopreservation procedures for research-grade human skin fibroblast lines. We also describe how to make feeder layers for hESCs using these fibroblasts. Copyright © 2016 John Wiley & Sons, Inc.

  12. Polymorphisms in voltage-gated sodium channel gene and susceptibility of Aedes albopictus to insecticides in three districts of northern West Bengal, India.

    PubMed

    Chatterjee, Moytrey; Ballav, Sudeep; Maji, Ardhendu K; Basu, Nandita; Sarkar, Biplab Chandra; Saha, Pabitra

    2018-01-01

    The control and prevention of dengue largely depends on vector control measures, environmental management, and personal protection. Dengue control programmes are facing great challenges due to development of insecticide resistance among vector mosquitoes. Information on susceptibility status to different insecticides is important for national programmes to formulate vector control strategies. We have studied the larval susceptibility of Aedes albopictus to temephos and adult susceptibility to 4% DDT, 0.05% deltamethrin, and 5% malathion as per WHO protocols in the northern districts of West Bengal. Polymorphisms in the VGSC gene were studied by direct sequencing of PCR products. The Ae. albopictus larval population showed sensitive [Resistance Ratio (RR99)<3] to moderate levels of resistance (510) to temephos at different study sites. Adult bioassay results revealed that Ae. albopictus was highly resistant to DDT [Corrected Mortality (CM) < 90%] in all the study sites and susceptible to deltamethrin and malathion (CM > 98%), except in Dhupguri where a low level of resistance to deltamethrin (CM = 96.25%) was recorded. None of the six important kdr mutations (S953P, I975M/V, L978, V980G, F1474C, D1703Y) were found in the VGSC of studied mosquitoes, but we identified 11 synonymous and 1 non-synonymous mutation in the VGSC gene. The higher susceptibility level to deltamethrin and malathion, along with the absence of important kdr mutations indicates that these two insecticides are still effective against Ae. albopictus in the study areas. The susceptibility status of temephos should be monitored closely as low to moderate levels of resistance were observed in few sites. A similar study is recommended for monitoring and early detection of insecticide resistance in other parts of the country.

  13. Polymorphisms in voltage-gated sodium channel gene and susceptibility of Aedes albopictus to insecticides in three districts of northern West Bengal, India

    PubMed Central

    Chatterjee, Moytrey; Ballav, Sudeep; Maji, Ardhendu K.; Basu, Nandita; Sarkar, Biplab Chandra

    2018-01-01

    Background The control and prevention of dengue largely depends on vector control measures, environmental management, and personal protection. Dengue control programmes are facing great challenges due to development of insecticide resistance among vector mosquitoes. Information on susceptibility status to different insecticides is important for national programmes to formulate vector control strategies. Methods We have studied the larval susceptibility of Aedes albopictus to temephos and adult susceptibility to 4% DDT, 0.05% deltamethrin, and 5% malathion as per WHO protocols in the northern districts of West Bengal. Polymorphisms in the VGSC gene were studied by direct sequencing of PCR products. Results The Ae. albopictus larval population showed sensitive [Resistance Ratio (RR99)<3] to moderate levels of resistance (510) to temephos at different study sites. Adult bioassay results revealed that Ae. albopictus was highly resistant to DDT [Corrected Mortality (CM) < 90%] in all the study sites and susceptible to deltamethrin and malathion (CM > 98%), except in Dhupguri where a low level of resistance to deltamethrin (CM = 96.25%) was recorded. None of the six important kdr mutations (S953P, I975M/V, L978, V980G, F1474C, D1703Y) were found in the VGSC of studied mosquitoes, but we identified 11 synonymous and 1 non-synonymous mutation in the VGSC gene. Conclusion The higher susceptibility level to deltamethrin and malathion, along with the absence of important kdr mutations indicates that these two insecticides are still effective against Ae. albopictus in the study areas. The susceptibility status of temephos should be monitored closely as low to moderate levels of resistance were observed in few sites. A similar study is recommended for monitoring and early detection of insecticide resistance in other parts of the country. PMID:29309419

  14. Establishment of immortalized human erythroid progenitor cell lines able to produce enucleated red blood cells.

    PubMed

    Kurita, Ryo; Suda, Noriko; Sudo, Kazuhiro; Miharada, Kenichi; Hiroyama, Takashi; Miyoshi, Hiroyuki; Tani, Kenzaburo; Nakamura, Yukio

    2013-01-01

    Transfusion of red blood cells (RBCs) is a standard and indispensable therapy in current clinical practice. In vitro production of RBCs offers a potential means to overcome a shortage of transfusable RBCs in some clinical situations and also to provide a source of cells free from possible infection or contamination by microorganisms. Thus, in vitro production of RBCs may become a standard procedure in the future. We previously reported the successful establishment of immortalized mouse erythroid progenitor cell lines that were able to produce mature RBCs very efficiently. Here, we have developed a reliable protocol for establishing immortalized human erythroid progenitor cell lines that are able to produce enucleated RBCs. These immortalized cell lines produce functional hemoglobin and express erythroid-specific markers, and these markers are upregulated following induction of differentiation in vitro. Most importantly, these immortalized cell lines all produce enucleated RBCs after induction of differentiation in vitro, although the efficiency of producing enucleated RBCs remains to be improved further. To the best of our knowledge, this is the first demonstration of the feasibility of using immortalized human erythroid progenitor cell lines as an ex vivo source for production of enucleated RBCs.

  15. Efficacy of ovitrap colors and patterns for attracting Aedes albopictus at suburban field sites in North-Central Florida

    USDA-ARS?s Scientific Manuscript database

    We sought to visually enhance the attractiveness of a standard black ovitrap routinely used in surveillance of the Asian Tiger Mosquito, Aedes albopictus, and now being used as lethal ovitraps in Aedes aegypti dengue control programs. Black plastic drinking cups (ovitraps) were visually altered to ...

  16. Establishment and characterization of a novel osteosarcoma cell line: CHOS.

    PubMed

    Liu, Yunlu; Feng, Xiaobo; Zhang, Yukun; Jiang, Hongyan; Cai, Xianyi; Yan, Xinxin; Huang, Zengfa; Mo, Fengbo; Yang, Wen; Yang, Cao; Yang, Shuhua; Liu, Xianzhe

    2016-12-01

    Osteosarcoma has a well-recognized bimodal distribution, with the first peak in adolescence and another in the elderly age-group. The elderly patients have different clinical features and a poorer prognosis as compared to adolescents. To better understand the biological features of osteosarcoma in the elderly population, we established a new human osteosarcoma cell line from a 58-year-old man with primary chondroblastic osteosarcoma. After 6 months of continuous culture in vitro for over 50 passages, an immortalized cell line CHOS was established. The cell line was well-characterized by cytogenetic, biomarker, functional, and histological analyses. The CHOS cells exhibited a spindle-shaped morphology and a doubling time of 36 h. Cytogenetic analysis of CHOS cells revealed the loss of chromosome Y and the gain of chromosome 12. Quantitative real-time polymerase chain reaction (RT-PCR), Western blotting and/or immunofluorescence revealed the expression of chondroblastic, mesenchymal and tumor metastasis markers in the CHOS cells. Compared with the osteosarcoma cell line, the CHOS cells were found to be more sensitive to cisplatin and doxorubicin, but were resistant to methotrexate. The cell line was highly tumorigenic and maintained the histological characteristics and invasive nature of the original tumor. Furthermore, on immunohistochemical analysis, the xenografts and metastases were found to co-express collagen II, aggrecan, vimentin and S100A4 that resembled the original tumor cells. Our results indicate, the potential of CHOS cell line to serve as a useful tool for further studies on the molecular biology of osteosarcoma, especially in the elderly patients. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:2116-2125, 2016. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  17. Establishment and characterization of five immortalized human scalp dermal papilla cell lines.

    PubMed

    Kwack, Mi Hee; Yang, Jung Min; Won, Gong Hee; Kim, Moon Kyu; Kim, Jung Chul; Sung, Young Kwan

    2018-02-05

    Dermal papilla (DP) regulates the growth and cycling of hair follicles. Cultured DP cells are useful for the study of their role in relation to hair growth and regeneration. However, cultivation of human DP cells is tedious and difficult. In addition, cultured DP cells possess a relatively short replicative life span, requiring immortalized human DP cell lines. We previously established an immortalized human DP cell line, SV40T-hTERT-DPC, by introducing human telomerase reverse transcriptase (hTERT) gene into the transformed cell line, SV40T-DPC. In this study, we co-transfected the simian virus 40 large T antigen (SV40T-Ag) and hTERT into DP cells from scalp hair follicles from a male with androgenetic alopecia and established five immortalized DP cell lines and named KNU-101, KNU-102, KNU-103, KNU-201 and KNU-202. We then evaluated tumorigenicity, expression of DP markers, responses to androgen, Wnt3a and BMP4, and expression of DP signature genes. These cell lines displayed early passage morphology and maintained responses to androgen, Wnt and BMP. Furthermore, these cell lines expressed DP markers and DP signature genes. KNU cell lines established in this study are potentially useful sources for hair research. Copyright © 2018 Elsevier Inc. All rights reserved.

  18. Comparative performance of fetal goat tongue cell line ZZ-R 127 and fetal porcine kidney cell line LFBK-αvβ6 for Foot-and-mouth disease virus isolation.

    PubMed

    Fukai, Katsuhiko; Morioka, Kazuki; Yamada, Manabu; Nishi, Tatsuya; Yoshida, Kazuo; Kitano, Rie; Yamazoe, Reiko; Kanno, Toru

    2015-07-01

    The fetal goat tongue cell line ZZ-R 127 and the fetal porcine kidney cell line LFBK-α(v)β(6) have been reported to have high sensitivity to various Foot-and-mouth disease virus (FMDV) strains. The suitability of ZZ-R 127 cells for FMDV isolation not only from epithelial suspensions but also from other clinical samples has already been confirmed in a previous study. However, to our knowledge, the suitability of LFBK-α(v)β(6) cells has not been evaluated using clinical samples other than epithelial materials. In addition, both cell lines have never been compared, in terms of use for FMDV isolation, under the same conditions. Therefore, in the current study, the virus isolation rates of both cell lines were compared using clinical samples collected from animals infected experimentally with FMDV. Viruses were successfully isolated from clinical samples other than epithelial suspensions for both cell lines. The virus isolation rates for the 2 cell lines were not significantly different. The Cohen kappa coefficients between the virus isolation results for both cell lines were significantly high. Taken together, these results confirmed the suitability of LFBK-α(v)β(6) cells for FMDV isolation from clinical samples other than epithelial suspensions. The levels of susceptibility of both cell lines to FMDV isolation were also confirmed to be almost the same. © 2015 The Author(s).

  19. Establishment of a novel human medulloblastoma cell line characterized by highly aggressive stem-like cells.

    PubMed

    Silva, Patrícia Benites Gonçalves da; Rodini, Carolina Oliveira; Kaid, Carolini; Nakahata, Adriana Miti; Pereira, Márcia Cristina Leite; Matushita, Hamilton; Costa, Silvia Souza da; Okamoto, Oswaldo Keith

    2016-08-01

    Medulloblastoma is a highly aggressive brain tumor and one of the leading causes of morbidity and mortality related to childhood cancer. These tumors display differential ability to metastasize and respond to treatment, which reflects their high degree of heterogeneity at the genetic and molecular levels. Such heterogeneity of medulloblastoma brings an additional challenge to the understanding of its physiopathology and impacts the development of new therapeutic strategies. This translational effort has been the focus of most pre-clinical studies which invariably employ experimental models using human tumor cell lines. Nonetheless, compared to other cancers, relatively few cell lines of human medulloblastoma are available in central repositories, partly due to the rarity of these tumors and to the intrinsic difficulties in establishing continuous cell lines from pediatric brain tumors. Here, we report the establishment of a new human medulloblastoma cell line which, in comparison with the commonly used and well-established cell line Daoy, is characterized by enhanced proliferation and invasion capabilities, stem cell properties, increased chemoresistance, tumorigenicity in an orthotopic metastatic model, replication of original medulloblastoma behavior in vivo, strong chromosome structural instability and deregulation of genes involved in neural development. These features are advantageous for designing biologically relevant experimental models in clinically oriented studies, making this novel cell line, named USP-13-Med, instrumental for the study of medulloblastoma biology and treatment.

  20. Mouse DRG Cell Line with Properties of Nociceptors.

    PubMed

    Doran, Ciara; Chetrit, Jonathan; Holley, Matthew C; Grundy, David; Nassar, Mohammed A

    2015-01-01

    In vitro cell lines from DRG neurons aid drug discovery because they can be used for early stage, high-throughput screens for drugs targeting pain pathways, with minimal dependence on animals. We have established a conditionally immortal DRG cell line from the Immortomouse. Using immunocytochemistry, RT-PCR and calcium microfluorimetry, we demonstrate that the cell line MED17.11 expresses markers of cells committed to the sensory neuron lineage. Within a few hours under differentiating conditions, MED17.11 cells extend processes and following seven days of differentiation, express markers of more mature DRG neurons, such as NaV1.7 and Piezo2. However, at least at this time-point, the nociceptive marker NaV1.8 is not expressed, but the cells respond to compounds known to excite nociceptors, including the TRPV1 agonist capsaicin, the purinergic receptor agonist ATP and the voltage gated sodium channel agonist, veratridine. Robust calcium transients are observed in the presence of the inflammatory mediators bradykinin, histamine and norepinephrine. MED17.11 cells have the potential to replace or reduce the use of primary DRG culture in sensory, pain and developmental research by providing a simple model to study acute nociception, neurite outgrowth and the developmental specification of DRG neurons.

  1. Characterization of immortalized human mammary epithelial cell line HMEC 2.6.

    PubMed

    Joshi, Pooja S; Modur, Vishnu; Cheng, JiMing; Robinson, Kathy; Rao, Krishna

    2017-10-01

    Primary human mammary epithelial cells have a limited life span which makes it difficult to study them in vitro for most purposes. To overcome this problem, we have developed a cell line that was immortalized using defined genetic elements, and we have characterized this immortalized non-tumorigenic human mammary epithelial cell line to establish it as a potential model system. human mammary epithelial cells were obtained from a healthy individual undergoing reduction mammoplasty at SIU School of Medicine. The cells were transduced with CDK4R24C followed by transduction with human telomerase reverse transcriptase. Post all manipulation, the cells displayed a normal cell cycle phase distribution and were near diploid in nature, which was confirmed by flow cytometry and karyotyping. In vitro studies showed that the cells were anchorage dependent and were non-invasive in nature. The cell line expressed basal epithelial markers such as cytokeratin 7, CD10, and p63 and was negative for the expression of estrogen receptor and progesterone receptor. Upon G-band karyotyping, the cell line displayed the presence of a few cytogenic abnormalities, including trisomy 20 and trisomy 7, which are also commonly present in other immortalized mammary cell lines. Furthermore, the benign nature of these cells was confirmed by multiple in vitro and in vivo experiments. Therefore, we think that this cell line could serve as a good model to understand the molecular mechanisms involved in the development and progression of breast cancer and to also assess the effect of novel therapeutics on human mammary epithelial cells.

  2. Susceptibility of Italian populations of Aedes albopictus to temephos and to other insecticides.

    PubMed

    Romi, Roberto; Toma, Luciano; Severini, Francesco; Di Luca, Marco

    2003-12-01

    We evaluated larval and adult susceptibility to 5 insecticides commonly used for control of Aedes albopictus, by using 15 field-collected populations from northern and central Italy, and we compared these data, collected in 2002, with those from evaluations conducted in 1992-93, 1996, and 1998-99. Larvae were tested for susceptibility to temephos and to the conventional diagnostic dosages of chlorpyrifos and fenthion proposed by the World Health Organization for Aedes aegypti. Adults were exposed to the diagnostic dosages of deltamethrin and permethrin. Overall, all of the populations tested in 2002 were still susceptible to temephos: median lethal concentration (LC50) values ranged from 0.0026 to 0.0085 mg/liter, and LC99 values ranged from 0.0093 to 0.023 mg/liter. These populations were also fully susceptible to chlorpyrifos and fenthion, and adults were susceptible to deltamethrin and permethrin. When comparing these data with those from previous evaluations, we observed a slight yet progressive increase in the LC50 and LC99 values of susceptibility to temephos, and these values are approaching the diagnostic dosage of 0.02 mg/liter proposed for Ae. aegypti. However, the implications of these findings need to be considered in light of the results of previous studies that have shown that larval Ae. albopictus are less susceptible than Ae. aegypti to organophosphate insecticides.

  3. DNA excision repair in cell extracts from human cell lines exhibiting hypersensitivity to DNA-damaging agents

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hansson, J.; Keyse, S.M.; Lindahl, T.

    Whole cell extracts from human lymphoid cell lines can perform in vitro DNA repair synthesis in plasmids damaged by agents including UV or cis-diamminedichloroplatinum(II) (cis-DDP). Extracts from xeroderma pigmentosum (XP) cells are defective in repair synthesis. We have now studied in vitro DNA repair synthesis using extracts from lymphoblastoid cell lines representing four human hereditary syndromes with increased sensitivity to DNA-damaging agents. Extracts of cell lines from individuals with the sunlight-sensitive disorders dysplastic nevus syndrome or Cockayne's syndrome (complementation groups A and B) showed normal DNA repair synthesis in plasmids with UV photoproducts. This is consistent with in vivo measurementsmore » of the overall DNA repair capacity in such cell lines. A number of extracts were prepared from two cell lines representing the variant form of XP (XP-V). Half of the extracts prepared showed normal levels of in vitro DNA repair synthesis in plasmids containing UV lesions, but the remainder of the extracts from the same cell lines showed deficient repair synthesis, suggesting the possibility of an unusually labile excision repair protein in XP-V. Fanconi's anemia (FA) cells show cellular hypersensitivity to cross-linking agents including cis-DDP. Extracts from cell lines belonging to two different complementation groups of FA showed normal DNA repair synthesis in plasmids containing cis-DDP or UV adducts. Thus, there does not appear to be an overall excision repair defect in FA, but the data do not exclude a defect in the repair of interstrand DNA cross-links.« less

  4. Anticancer effects of resveratrol in canine hemangiosarcoma cell lines.

    PubMed

    Carlson, A; Alderete, K S; Grant, M K O; Seelig, D M; Sharkey, L C; Zordoky, B N M

    2018-06-01

    Hemangiosarcoma (HSA) is a highly malignant tumour with aggressive biological behaviour. HSAs are more common in dogs than other domestic animals. The median survival time of dogs with HSA remains short, even with chemotherapy and surgery. Therefore, there is a critical need to improve the adjuvant chemotherapeutic regimens to improve clinical outcomes in dogs with HSA. Resveratrol has been shown to possess strong anti-proliferative and/or pro-apoptotic properties in human cancer cell lines. Nevertheless, the potential anticancer effects of resveratrol have not been reported in canine HSAs. The objective of this study is to determine the growth inhibitory effects of resveratrol in HSA cells when used alone or in combination with doxorubicin, a commonly used chemotherapeutic agent. Frog and DD-1 canine HSA cell lines were treated with varying concentrations of resveratrol with and without doxorubicin. Cell viability was measured by the MTT assay. The expression of apoptotic proteins, activation of p38 mitogen-activated protein kinase (MAPK), AMP-activated protein kinase (AMPK) and extracellular signal-regulated kinase 1/2 (ERK1/2) were assessed by western blotting. Similar to human cancer cell lines, resveratrol markedly inhibited the growth and induced apoptosis in both HSA cell lines. Mechanistically, resveratrol activated p38 MAPK, but did not affect the AMPK or the ERK1/2 pathways. Additional experiments showed that resveratrol augmented the growth-inhibitory and apoptotic effects of doxorubicin in both HSA cell lines. These findings suggest that resveratrol has pro-apoptotic effects in canine HSA cells; therefore, its use as a potential adjunct therapy in canine HSA patients warrants further investigation. © 2017 John Wiley & Sons Ltd.

  5. Modelling cell population growth with applications to cancer therapy in human tumour cell lines.

    PubMed

    Basse, Britta; Baguley, Bruce C; Marshall, Elaine S; Wake, Graeme C; Wall, David J N

    2004-01-01

    In this paper we present an overview of the work undertaken to model a population of cells and the effects of cancer therapy. We began with a theoretical one compartment size structured cell population model and investigated its asymptotic steady size distributions (SSDs) (On a cell growth model for plankton, MMB JIMA 21 (2004) 49). However these size distributions are not similar to the DNA (size) distributions obtained experimentally via the flow cytometric analysis of human tumour cell lines (data obtained from the Auckland Cancer Society Research Centre, New Zealand). In our one compartment model, size was a generic term, but in order to obtain realistic steady size distributions we chose size to be DNA content and devised a multi-compartment mathematical model for the cell division cycle where each compartment corresponds to a distinct phase of the cell cycle (J. Math. Biol. 47 (2003) 295). We then incorporated another compartment describing the possible induction of apoptosis (cell death) from mitosis phase (Modelling cell death in human tumour cell lines exposed to anticancer drug paclitaxel, J. Math. Biol. 2004, in press). This enabled us to compare our model to flow cytometric data of a melanoma cell line where the anticancer drug, paclitaxel, had been added. The model gives a dynamic picture of the effects of paclitaxel on the cell cycle. We hope to use the model to describe the effects of other cancer therapies on a number of different cell lines. Copyright 2004 Elsevier Ltd.

  6. Evaluating the Vector Control Potential of the In2Care® Mosquito Trap Against Aedes aegypti and Aedes albopictus Under Semifield Conditions in Manatee County, Florida.

    PubMed

    Buckner, Eva A; Williams, Katie F; Marsicano, Ambyr L; Latham, Mark D; Lesser, Christopher R

    2017-09-01

    Successful integrated vector management programs may need new strategies in addition to conventional larviciding and adulticiding strategies to target Aedes aegypti and Ae. albopictus, which can develop in small, often cryptic, artificial and natural containers. The In2Care® mosquito trap was recently developed to target and kill larval and adult stages of these invasive container-inhabiting Aedes mosquitoes by utilizing autodissemination. Gravid females that visit the trap pick up pyriproxyfen (PPF) that they later transfer to nearby larval habitats as well as Beauveria bassiana spores that slowly kill them. We assessed the efficacy of the In2Care mosquito trap in a semifield setting against locally sourced strains of Ae. aegypti and Ae. albopictus. We found that the In2Care mosquito trap is attractive to gravid Ae. aegypti and Ae. albopictus females and serves as an egg sink, preventing any adult emergence from the trap (P = 0.0053 for both species). Adult females successfully autodisseminated PPF to surrounding water-filled containers, leading to a statistically significant reduction in new mosquito emergence (P ≤ 0.0002 for both species). Additionally, we found effective contamination with Beauveria bassiana spores, which significantly reduced the survivorship of exposed Ae. aegypti and Ae. albopictus (P ≤ 0.008 for both species in all experimental setups). In summary, the In2Care mosquito trap successfully killed multiple life stages of 2 main mosquito vector species found in Florida under semifield conditions.

  7. Lung cancer cell lines: Useless artifacts or invaluable tools for medical science?

    PubMed Central

    Gazdar, Adi F.; Gao, Boning; Minna, John D.

    2011-01-01

    Multiple cell lines (estimated at 300–400) have been established from human small cell (SCLC) and non-small cell lung cancers (NSCLC). These cell lines have been widely dispersed to and used by the scientific community worldwide, with over 8000 citations resulting from their study. However, there remains considerable skepticism on the part of the scientific community as to the validity of research resulting from their use. These questions center around the genomic instability of cultured cells, lack of differentiation of cultured cells and absence of stromal–vascular–inflammatory cell compartments. In this report we discuss the advantages and disadvantages of the use of cell lines, address the issues of instability and lack of differentiation. Perhaps the most important finding is that every important, recurrent genetic and epigenetic change including gene mutations, deletions, amplifications, translocations and methylation-induced gene silencing found in tumors has been identified in cell lines and vice versa. These “driver mutations” represented in cell lines offer opportunities for biological characterization and application to translational research. Another potential shortcoming of cell lines is the difficulty of studying multistage pathogenesis in vitro.To overcome this problem, we have developed cultures from central and peripheral airways that serve as models for the multistage pathogenesis of tumors arising in these two very different compartments. Finally the issue of cell line contamination must be addressed and safeguarded against. A full understanding of the advantages and shortcomings of cell lines is required for the investigator to derive the maximum benefit from their use. PMID:20079948

  8. Establishment and characterization of a new human acinar cell carcinoma cell line, Faraz-ICR, from pancreas.

    PubMed

    Rezaei, Marzieh; Hosseini, Ahmad; Nikeghbalian, Saman; Ghaderi, Abbas

    Basic research in the field of acinar cell carcinoma (ACC) as a rare neoplasm of the pancreas is dependent on the availability of pragmatic model such as new pancreatic cancer cell lines. Thus, establishment and characterization of new pancreatic cancer cell lines from ACC origin are deemed important. Faraz-ICR cell line was derived from a 58-years old woman with pancreatic acinar cell carcinoma by the collagenase digestion protocol. We characterized the cell line by examining its morphology and cytostructural and functional profile. Faraz-ICR has a doubling time of 35 hours and grows in soft agar with a colony-forming efficiency of 25%. The cell had nearly normal pattern of chromosomes in karyotype analysis and Comparative Genomic Hybridization (CGH) array analysis. Evaluation of cells by flowcytometry showed that Faraz-ICR is negative for EpCAM and mesenchymal markers in different passages, and has epithelial nature. Immunofluorescence staining revealed that cells were strongly positive for vimentin, desmin, ezrin, S100, nestin and they were negative for pan-cytokeratins, chromogranin and alpha smooth muscle actin. We were able to establish a new pancreatic carcinoma cell line with partial aspects of Epithelial-mesenchymal transition and aggressiveness. This cell line might be suitable for studying various anticancer drugs and protein profile aiming to see any possible tumor associated marker for ACC. Copyright © 2017 IAP and EPC. Published by Elsevier B.V. All rights reserved.

  9. Low-dose non-targeted radiation effects in human esophageal adenocarcinoma cell lines.

    PubMed

    Hanu, Christine; Wong, Raimond; Sur, Ranjan K; Hayward, Joseph E; Seymour, Colin; Mothersill, Carmel

    2017-02-01

    To investigate non-targeted radiation effects in esophageal adenocarcinoma cell lines (OE19 and OE33) using human keratinocyte and colorectal cancer cell reporters following γ-ray exposure. Both clonogenic assays and ratiometric calcium endpoints were used to check for the occurrence of bystander signals in reporter cells. We report data suggesting that γ-irradiation increases cell killing over the expected linear quadratic (LQ) model levels in the OE19 cell line exposed to doses below 1 Gy, i.e. which may be suggestive to be a low hyper-radiosensitive (HRS) response to direct irradiation. Both EAC cell lines (OE19 and OE33) have the ability to produce bystander signals when irradiated cell conditioned medium (ICCM) is placed onto human keratinocyte reporters, but do not seem to be capable of responding to bystander signals when placed on their autologous reporters. Further work with human keratinocyte reporter models showed statistically significant intracellular calcium fluxes following exposure of the reporters to ICCM harvested from both EAC cell lines exposed to 0.5 Gy. These experiments suggest that the OE19 and OE33 cell lines produce bystander signals in human keratinocyte reporter cells. However, the radiosensitivity of the EAC cell lines used in this study cannot be enhanced by the bystander response since both cell lines could not respond to bystander signals.

  10. Functional characterization of cell hybrids generated by induced fusion of primary porcine mesenchymal stem cells with an immortal murine cell line.

    PubMed

    Islam, M Q; Ringe, J; Reichmann, E; Migotti, R; Sittinger, M; da S Meirelles, L; Nardi, N B; Magnusson, P; Islam, K

    2006-10-01

    Bone marrow mesenchymal stem cells (MSC) integrate into various organs and contribute to the regeneration of diverse tissues. However, the mechanistic basis of the plasticity of MSC is not fully understood. The change of cell fate has been suggested to occur through cell fusion. We have generated hybrid cell lines by polyethylene-glycol-mediated cell fusion of primary porcine MSC with the immortal murine fibroblast cell line F7, a derivative of the GM05267 cell line. The hybrid cell lines display fibroblastic morphology and proliferate like immortal cells. They contain tetraploid to hexaploid porcine chromosomes accompanied by hypo-diploid murine chromosomes. Interestingly, many hybrid cell lines also express high levels of tissue-nonspecific alkaline phosphatase, which is considered to be a marker of undifferentiated embryonic stem cells. All tested hybrid cell lines retain osteogenic differentiation, a few of them also retain adipogenic potential, but none retain chondrogenic differentiation. Conditioned media from hybrid cells enhance the proliferation of both early-passage and late-passage porcine MSC, indicating that the hybrid cells secrete diffusible growth stimulatory factors. Murine F7 cells thus have the unique property of generating immortal cell hybrids containing unusually high numbers of chromosomes derived from normal cells. These hybrid cells can be employed in various studies to improve our understanding of regenerative biology. This is the first report, to our knowledge, describing the generation of experimentally induced cell hybrids by using normal primary MSC.

  11. Risk Factors for the Presence of Aedes aegypti and Aedes albopictus in Domestic Water-Holding Containers in Areas Impacted by the Nam Theun 2 Hydroelectric Project, Laos

    PubMed Central

    Hiscox, Alexandra; Kaye, Angela; Vongphayloth, Khamsing; Banks, Ian; Piffer, Michele; Khammanithong, Phasouk; Sananikhom, Pany; Kaul, Surinder; Hill, Nigel; Lindsay, Steven W.; Brey, Paul T.

    2013-01-01

    We assessed risk factors for vectors of dengue and chikungunya viruses near a new hydroelectric project, Nam Theun 2, in Laos. Immature stages of Aedes aegypti were found only in sites within 40 km of the urban provincial capital, but Aedes albopictus was found throughout. Aedes aegypti pupae were most common in water storage jars (odds ratio [OR] = 4.72) and tires (OR = 2.99), and Ae. albopictus pupae were associated with tires in 2009 (OR = 10.87) and drums, tires, and jars in 2010 (drums OR = 3.05; tires OR = 3.45, jars OR = 6.59). Compared with water storage vessels, containers used for hygiene, cooking, and drinking were 80% less likely to harbor Ae. albopictus pupae in 2010 (OR = 0.20), and discarded waste was associated with a 3.64 increased odds of infestation. Vector control efforts should focus on source reduction of water storage containers, particularly concrete jars and tires. PMID:23458958

  12. Generation of stable PDX derived cell lines using conditional reprogramming.

    PubMed

    Borodovsky, Alexandra; McQuiston, Travis J; Stetson, Daniel; Ahmed, Ambar; Whitston, David; Zhang, Jingwen; Grondine, Michael; Lawson, Deborah; Challberg, Sharon S; Zinda, Michael; Pollok, Brian A; Dougherty, Brian A; D'Cruz, Celina M

    2017-12-06

    Efforts to develop effective cancer therapeutics have been hindered by a lack of clinically predictive preclinical models which recapitulate this complex disease. Patient derived xenograft (PDX) models have emerged as valuable tools for translational research but have several practical limitations including lack of sustained growth in vitro. In this study, we utilized Conditional Reprogramming (CR) cell technology- a novel cell culture system facilitating the generation of stable cultures from patient biopsies- to establish PDX-derived cell lines which maintain the characteristics of the parental PDX tumor. Human lung and ovarian PDX tumors were successfully propagated using CR technology to create stable explant cell lines (CR-PDX). These CR-PDX cell lines maintained parental driver mutations and allele frequency without clonal drift. Purified CR-PDX cell lines were amenable to high throughput chemosensitivity screening and in vitro genetic knockdown studies. Additionally, re-implanted CR-PDX cells proliferated to form tumors that retained the growth kinetics, histology, and drug responses of the parental PDX tumor. CR technology can be used to generate and expand stable cell lines from PDX tumors without compromising fundamental biological properties of the model. It offers the ability to expand PDX cells in vitro for subsequent 2D screening assays as well as for use in vivo to reduce variability, animal usage and study costs. The methods and data detailed here provide a platform to generate physiologically relevant and predictive preclinical models to enhance drug discovery efforts.

  13. Reintroduction of the invasive mosquito species Aedes albopictus in Belgium in July 2013

    PubMed Central

    Boukraa, Slimane; Raharimalala, Fara N.; Zimmer, Jean-Yves; Schaffner, Francis; Bawin, Thomas; Haubruge, Eric; Francis, Frédéric

    2013-01-01

    Since its first report in 2000, the invasive mosquito Aedes albopictus was not found any more during the different entomological inspections performed at its place of introduction in Belgium between 2001 and 2012. In July 2013, one adult male was captured at the same site (a platform of imported used tires located in Vrasene, Oost-Vlaanderen Province), during a monitoring using CO2-baited trap. This finding suggests the reintroduction of the species in Belgium via the used tire trade. PMID:24325893

  14. Assessment of citalopram and escitalopram on neuroblastoma cell lines: Cell toxicity and gene modulation

    PubMed Central

    Sakka, Laurent; Delétage, Nathalie; Chalus, Maryse; Aissouni, Youssef; Sylvain-Vidal, Valérie; Gobron, Stéphane; Coll, Guillaume

    2017-01-01

    Selective serotonin reuptake inhibitors (SSRI) are common antidepressants which cytotoxicity has been assessed in cancers notably colorectal carcinomas and glioma cell lines. We assessed and compared the cytotoxicity of 2 SSRI, citalopram and escitalopram, on neuroblastoma cell lines. The study was performed on 2 non-MYCN amplified cell lines (rat B104 and human SH-SY5Y) and 2 human MYCN amplified cell lines (IMR32 and Kelly). Citalopram and escitalopram showed concentration-dependent cytotoxicity on all cell lines. Citalopram was more cytotoxic than escitalopram. IMR32 was the most sensitive cell line. The absence of toxicity on human primary Schwann cells demonstrated the safety of both molecules for myelin. The mechanisms of cytotoxicity were explored using gene-expression profiles and quantitative real-time PCR (qPCR). Citalopram modulated 1 502 genes and escitalopram 1 164 genes with a fold change ≥ 2. 1 021 genes were modulated by both citalopram and escitalopram; 481 genes were regulated only by citalopram while 143 genes were regulated only by escitalopram. Citalopram modulated 69 pathways (KEGG) and escitalopram 42. Ten pathways were differently modulated by citalopram and escitalopram. Citalopram drastically decreased the expression of MYBL2, BIRC5 and BARD1 poor prognosis factors of neuroblastoma with fold-changes of -107 (p<2.26 10−7), -24.1 (p<5.6 10−9) and -17.7 (p<1.2 10−7). CCNE1, AURKA, IGF2, MYCN and ERBB2 were more moderately down-regulated by both molecules. Glioma markers E2F1, DAPK1 and CCND1 were down-regulated. Citalopram displayed more powerful action with broader and distinct spectrum of action than escitalopram. PMID:28467792

  15. Assessment of citalopram and escitalopram on neuroblastoma cell lines. Cell toxicity and gene modulation.

    PubMed

    Sakka, Laurent; Delétage, Nathalie; Chalus, Maryse; Aissouni, Youssef; Sylvain-Vidal, Valérie; Gobron, Stéphane; Coll, Guillaume

    2017-06-27

    Selective serotonin reuptake inhibitors (SSRI) are common antidepressants which cytotoxicity has been assessed in cancers notably colorectal carcinomas and glioma cell lines. We assessed and compared the cytotoxicity of 2 SSRI, citalopram and escitalopram, on neuroblastoma cell lines. The study was performed on 2 non-MYCN amplified cell lines (rat B104 and human SH-SY5Y) and 2 human MYCN amplified cell lines (IMR32 and Kelly). Citalopram and escitalopram showed concentration-dependent cytotoxicity on all cell lines. Citalopram was more cytotoxic than escitalopram. IMR32 was the most sensitive cell line. The absence of toxicity on human primary Schwann cells demonstrated the safety of both molecules for myelin. The mechanisms of cytotoxicity were explored using gene-expression profiles and quantitative real-time PCR (qPCR). Citalopram modulated 1 502 genes and escitalopram 1 164 genes with a fold change ≥ 2. 1 021 genes were modulated by both citalopram and escitalopram; 481 genes were regulated only by citalopram while 143 genes were regulated only by escitalopram. Citalopram modulated 69 pathways (KEGG) and escitalopram 42. Ten pathways were differently modulated by citalopram and escitalopram. Citalopram drastically decreased the expression of MYBL2, BIRC5 and BARD1 poor prognosis factors of neuroblastoma with fold-changes of -107 (p<2.26 10-7), -24.1 (p<5.6 10-9) and -17.7 (p<1.2 10-7). CCNE1, AURKA, IGF2, MYCN and ERBB2 were more moderately down-regulated by both molecules. Glioma markers E2F1, DAPK1 and CCND1 were down-regulated. Citalopram displayed more powerful action with broader and distinct spectrum of action than escitalopram.

  16. Effect of application rate and persistence of boric acid sugar baits applied to plants control of Aedes albopictus

    USDA-ARS?s Scientific Manuscript database

    The use of toxic baits to kill adult Aedes albopictus (Skuse) mosquitoes is a safe and potentially effective alternative to the use of synthetic chemical insecticides. This study was made to identify effective application rates for boric acid-sugar solution baits sprayed onto plant surfaces and to ...

  17. [The characters and specific features of new human embryonic stem cells lines].

    PubMed

    Krylova, T A; Kol'tsova, A M; Zenin, V V; Gordeeva, O F; Musorina, A S; Goriachaia, T S; Shlykova, S A; Kamenetskaia, Iu K; Pinaev, G P; Polianskaia, G G

    2009-01-01

    Four continuous human embryonic stem cell lines (SC1, SC2, SC3 and SC4), derived from the blastocysts has been described. The cell lines were cultivated on mitotically inactivated human feeder cells. The cell lines SC1 and SC2 have passed through 150 population doublings and the cell lines SC3 and SC4 -- near 120 populations doublings, which exceeds Hayflick limit sufficiently. These cell lines maintain high activity of alkaline phosphatase, expression of transcription factor OCT-4 and cell surface antigens (SSEA-4, TRA-1-60 and TRA-1-81), confirming their ESC status and human specificity. Immunofluorescent detection of antigens, characteristic of ectoderm, endoderm and mesoderm confirms the ability of these cells to retain their pluripotency under in vitro condition. PCR analysis revealed expression of six genes specific for pluripotent cells (OCT-4, NANOG, DPPA3/STELLA, TDGF/CRIPTO and LEFTYA). Correlation between the level of proliferative activity and the character of DNA-bound fluorescent staining was found. Fluorescent dyes, Hoechst 33342 and PI, produced diffuse staining of the nuclei in slowly proliferating cells of the SC1 and SC2 lines. In contrast, in actively proliferating cells of the SC3 and SC4 lines, the clear staining of the nuclei was observed. Upon changing the cultivation condition, proliferative activity of SC3 and SC4 lines decreased and became similar to that of SC1 and SC2 lines. The character of the fluorescent staining of all these lines was also shown to be similar. These results show that quality of the fluorescent staining with Hoechst 33342 and PI reflects the level of proliferation. Possible causes and mechanisms of this feature of human ESC are discussed.

  18. The effect of interspecific competition on the temporal dynamics of Aedes albopictus and Culex pipiens.

    PubMed

    Marini, Giovanni; Guzzetta, Giorgio; Baldacchino, Frederic; Arnoldi, Daniele; Montarsi, Fabrizio; Capelli, Gioia; Rizzoli, Annapaola; Merler, Stefano; Rosà, Roberto

    2017-02-23

    Aedes albopictus and Culex pipiens larvae reared in the same breeding site compete for resources, with an asymmetrical outcome that disadvantages only the latter species. The impact of these interactions on the overall ecology of these two species has not yet been assessed in the natural environment. In the present study, the temporal patterns of adult female mosquitoes from both species were analysed in north-eastern Italy, and substantial temporal shifts between abundance curves of Cx. pipiens and Ae. albopictus were observed in several sites. To understand which factors can drive the observed temporal shifts, we developed a mechanistic model that takes explicitly into account the effect of temperature on the development and survival of all mosquito stages. We also included into the model the effect of asymmetric interspecific competition, by adding a mortality term for Cx. pipiens larvae proportional to the larval abundance of Ae. albopictus within the same breeding site. Model calibration was performed through a Markov Chain Monte Carlo approach using weekly capture data collected in our study sites during 2014 and 2015. In almost half of observation sites, temporal shifts were due to competition, with an early decline of Cx. pipiens caused by the concurrent rise in abundance of its competitor, and this effect was enhanced by higher abundance of both species. We estimate that competition may reduce Cx. pipiens abundance in some sites by up to about 70%. However, in some cases temporal shifts can also be explained in the absence of competition between species resulting from a "temporal niche" effect, when the optimal fitness to environmental conditions for the two species are reached at different times of the year. Our findings demonstrate the importance of considering ecological interactions and, in particular, competition between mosquito species in temperate climates, with important implications for risk assessment of mosquito transmitted pathogens, as well

  19. Genetic heterogeneity of RPMI-8402, a T-acute lymphoblastic leukemia cell line

    PubMed Central

    STOCZYNSKA-FIDELUS, EWELINA; PIASKOWSKI, SYLWESTER; PAWLOWSKA, ROZA; SZYBKA, MALGORZATA; PECIAK, JOANNA; HULAS-BIGOSZEWSKA, KRYSTYNA; WINIECKA-KLIMEK, MARTA; RIESKE, PIOTR

    2016-01-01

    Thorough examination of genetic heterogeneity of cell lines is uncommon. In order to address this issue, the present study analyzed the genetic heterogeneity of RPMI-8402, a T-acute lymphoblastic leukemia (T-ALL) cell line. For this purpose, traditional techniques such as fluorescence in situ hybridization and immunocytochemistry were used, in addition to more advanced techniques, including cell sorting, Sanger sequencing and massive parallel sequencing. The results indicated that the RPMI-8402 cell line consists of several genetically different cell subpopulations. Furthermore, massive parallel sequencing of RPMI-8402 provided insight into the evolution of T-ALL carcinogenesis, since this cell line exhibited the genetic heterogeneity typical of T-ALL. Therefore, the use of cell lines for drug testing in future studies may aid the progress of anticancer drug research. PMID:26870252

  20. Establishment of optimized MDCK cell lines for reliable efflux transport studies.

    PubMed

    Gartzke, Dominik; Fricker, Gert

    2014-04-01

    Madin-Darby canine kidney (MDCK) cells transfected with human MDR1 gene (MDCK-MDR1) encoding for P-glycoprotein (hPgp, ABCB1) are widely used for transport studies to identify drug candidates as substrates of this efflux protein. Therefore, it is necessary to rely on constant and comparable expression levels of Pgp to avoid false negative or positive results. We generated a cell line with homogenously high and stable expression of hPgp through sorting single clones from a MDCK-MDR1 cell pool using fluorescence-activated cell sorting (FACS). To obtain control cell lines for evaluation of cross-interactions with endogenous canine Pgp (cPgp) wild-type cells were sorted with a low expression pattern of cPgp in comparison with the MDCK-MDR1. Expression of other transporters was also characterized in both cell lines by quantitative real-time PCR and Western blot. Pgp function was investigated applying the Calcein-AM assay as well as bidirectional transport assays using (3) H-Digoxin, (3) H-Vinblastine, and (3) H-Quinidine as substrates. Generated MDCK-MDR1 cell lines showed high expression of hPgp. Control MDCK-WT cells were optimized in showing a comparable expression level of cPgp in comparison with MDCK-MDR1 cell lines. Generated cell lines showed higher and more selective Pgp transport compared with parental cells. Therefore, they provide a significant improvement in the performance of efflux studies yielding more reliable results. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  1. Global gene expression analyses of hematopoietic stem cell-like cell lines with inducible Lhx2 expression

    PubMed Central

    Richter, Karin; Wirta, Valtteri; Dahl, Lina; Bruce, Sara; Lundeberg, Joakim; Carlsson, Leif; Williams, Cecilia

    2006-01-01

    Background Expression of the LIM-homeobox gene Lhx2 in murine hematopoietic cells allows for the generation of hematopoietic stem cell (HSC)-like cell lines. To address the molecular basis of Lhx2 function, we generated HSC-like cell lines where Lhx2 expression is regulated by a tet-on system and hence dependent on the presence of doxycyclin (dox). These cell lines efficiently down-regulate Lhx2 expression upon dox withdrawal leading to a rapid differentiation into various myeloid cell types. Results Global gene expression of these cell lines cultured in dox was compared to different time points after dox withdrawal using microarray technology. We identified 267 differentially expressed genes. The majority of the genes overlapping with HSC-specific databases were those down-regulated after turning off Lhx2 expression and a majority of the genes overlapping with those defined as late progenitor-specific genes were the up-regulated genes, suggesting that these cell lines represent a relevant model system for normal HSCs also at the level of global gene expression. Moreover, in situ hybridisations of several genes down-regulated after dox withdrawal showed overlapping expression patterns with Lhx2 in various tissues during embryonic development. Conclusion Global gene expression analysis of HSC-like cell lines with inducible Lhx2 expression has identified genes putatively linked to self-renewal / differentiation of HSCs, and function of Lhx2 in organ development and stem / progenitor cells of non-hematopoietic origin. PMID:16600034

  2. Development and characterization of a cell line WAF from freshwater shark Wallago attu.

    PubMed

    Dubey, Akhilesh; Goswami, Mukunda; Yadav, Kamalendra; Sharma, Bhagwati S

    2014-02-01

    A new epithelial cell line, WAF was developed from caudal fin of freshwater shark, Wallago attu. The cell line was optimally maintained at 28 °C in Leibovitz-15 (L-15) medium supplemented with 20 % fetal bovine serum. The cell line was characterized by various cytogenetic and molecular markers. The cytogenetic analysis revealed a diploid count of 86 chromosomes at different passages. The origin of the cell lines was confirmed by the amplification of 547 and 654 bp sequences of 16S rRNA and cytochrome oxidase subunit I genes of mitochondrial DNA, respectively. WAF cells were characterized for their growth characteristics at different temperature and serum concentration. Epithelial morphology of the cell line was confirmed using immunocytochemistry. Further cell plating efficiency, transfection efficiency and viability of cryopreserved WAF cells was also determined. Cytotoxicity and genotoxicity assessment of cadmium salts on WAF cells by MTT, NR and comet assay illustrated the utility of this cell line as an in vitro model for aquatic toxicological studies. The cell line will be further useful for studying oxidative stress markers against aquatic pollutants.

  3. Different toxic effects of YTX in tumor K-562 and lymphoblastoid cell lines

    PubMed Central

    Fernández-Araujo, Andrea; Sánchez, Jon A.; Alfonso, Amparo; Vieytes, Mercedes R.; Botana, Luis M.

    2015-01-01

    Yessotoxin (YTX) modulates cellular phosphodiesterases (PDEs). In this regard, opposite effects had been described in the tumor model K-562 cell line and fresh human lymphocytes in terms of cell viability, cyclic adenosine 3',5'-cyclic monophosphate (cAMP) production and protein expression after YTX treatment. Studies in depth of the pathways activated by YTX in K-562 cell line, have demonstrated the activation of two different cell death types, apoptosis, and autophagy after 24 and 48 h of treatment, respectively. Furthermore, the key role of type 4A PDE (PDE4A) in both pathways activated by YTX was demonstrated. Therefore, taking into account the differences between cellular lines and fresh cells, a study of cell death pathways activated by YTX in a non-tumor cell line with mitotic activity, was performed. The cellular model used was the lymphoblastoid cell line that represents a non-tumor model with normal apoptotic and mitotic machinery. In this context, cell viability and cell proliferation, expression of proteins involved in cell death activated by YTX and mitochondrial mass, were studied after the incubation with the toxin. Opposite to the tumor model, no cell death activation was observed in lymphoblastoid cell line in the presence of YTX. In this sense, variations in apoptosis hallmarks were not detected in the lymphoblastoid cell line after YTX incubation, whereas this type I of programmed cell death was observed in K-562 cells. On the other hand, autophagy cell death was triggered in this cellular line, while other autophagic process is suggested in lymphoblastoid cells. These YTX effects are related to PDE4A in both cellular lines. In addition, while cell death is triggered in K-562 cells after YTX treatment, in lymphoblastoid cells the toxin stops cellular proliferation. These results point to YTX as a specific toxic compound of tumor cells, since in the non-tumor lymphoblastoid cell line, no cell death hallmarks are observed. PMID:26136685

  4. Hematopoietic Cancer Cell Lines Can Support Replication of Sabin Poliovirus Type 1

    PubMed Central

    van Eikenhorst, Gerco; de Gruijl, Tanja D.; van der Pol, Leo A.; Bakker, Wilfried A. M.

    2015-01-01

    Viral vaccines can be produced in adherent or in suspension cells. The objective of this work was to screen human suspension cell lines for the capacity to support viral replication. As the first step, it was investigated whether poliovirus can replicate in such cell lines. Sabin poliovirus type 1 was serially passaged on five human cell lines, HL60, K562, KG1, THP-1, and U937. Sabin type 1 was capable of efficiently replicating in three cell lines (K562, KG1, and U937), yielding high viral titers after replication. Expression of CD155, the poliovirus receptor, did not explain susceptibility to replication, since all cell lines expressed CD155. Furthermore, we showed that passaged virus replicated more efficiently than parental virus in KG1 cells, yielding higher virus titers in the supernatant early after infection. Infection of cell lines at an MOI of 0.01 resulted in high viral titers in the supernatant at day 4. Infection of K562 with passaged Sabin type 1 in a bioreactor system yielded high viral titers in the supernatant. Altogether, these data suggest that K562, KG1, and U937 cell lines are useful for propagation of poliovirus. PMID:25815312

  5. Efficient production of a gene mutant cell line through integrating TALENs and high-throughput cell cloning.

    PubMed

    Sun, Changhong; Fan, Yu; Li, Juan; Wang, Gancheng; Zhang, Hanshuo; Xi, Jianzhong Jeff

    2015-02-01

    Transcription activator-like effectors (TALEs) are becoming powerful DNA-targeting tools in a variety of mammalian cells and model organisms. However, generating a stable cell line with specific gene mutations in a simple and rapid manner remains a challenging task. Here, we report a new method to efficiently produce monoclonal cells using integrated TALE nuclease technology and a series of high-throughput cell cloning approaches. Following this method, we obtained three mTOR mutant 293T cell lines within 2 months, which included one homozygous mutant line. © 2014 Society for Laboratory Automation and Screening.

  6. Area-wide management of Aedes albopictus: choice of study sites based on geospatial characteristics, socioeconomic factors, and mosquito populations

    USDA-ARS?s Scientific Manuscript database

    Aedes (Stegomyia) albopictus (Skuse), the Asian tiger mosquito, is an introduced invasive species in the U.S. responsible for a significant proportion of service requests to local mosquito control programs. This container-utilizing mosquito is refractory to standard mosquito abatement measures in th...

  7. Nucleotide composition analysis of tRNA from leukemia patient cell samples and human cell lines.

    PubMed Central

    Agris, P F

    1975-01-01

    A technique developed for analysis of less than microgram quantities of tRNA has been applied to the study of human leukemia. Leucocytes from peripheal blood and bone marrow samples of six, untreated leukemia patients and cells of five different established human cell lines were maintained for 18 hours in media containing (32P)-phosphate. Incorporation of radioactive phosphate into the cells from the patient samples was slightly less than that of the cell lines. Likewise, incorporation of (32P)-phosphate into the tRNA of the patient samples (approximately 5 x 106 DPM/mug tRNA) was also less then that incorporated into the tRNA of the cell lines. The major and minor nucleotide compositions of the unfractionated tRNA preparations from each patient sample and each cell line were determined and compared. Similarities and differences in the major and minor nucleotide compositions of the tRNA preparations are discussed with reference to types of leukemia and the importance of patient sample analysis versus analysis of cultured human cells. PMID:1057159

  8. Survey on aedes mosquito density and pattern distribution of aedes aegypti and aedes albopictus in high and low incidence districts in north sumatera province

    NASA Astrophysics Data System (ADS)

    Siregar, Fazidah A.; Makmur, Tri

    2018-03-01

    Transmission and control of dengue hemorrhagic fever are related to its vectors. This study investigated vector density and distribution patterns of Aedes aegypty and Aedes albopictus in Medan and Langkat as high and low incidence district, respectively. An entomological survey was carried out in 304 households both in Medan and Langkat. The results showed that adult Ae. aegypti were predominantly in Medan, while adult Ae. albopictus was only in Langkat. Larvae indices (HI, CI, BI) for Aedes in Medan ( 35,13 and 43) were higher than langkat ( 22,8 and 30). Adult indices (AHI, AD, RR) for Ae. aegypti in Medan and for Ae. albopictus in Langkat were 20,38,24 and 3,5, and 5, respectively. Pattern distribution of Aedes larvae and adult mosquitoes in both district had similar pattern. Aedes larval indices and adult indices both in HIDs and LIDs were above the critical level, indicating potential high risk for DHF transmission. By multiple regression analysis, HI is predictor for DHF transmission in North Sumatera. Thus, in designing an effective control measures for dengue hemorrhagic fever, monitoring distribution and vector density is crucial.

  9. Novel antiproliferative flavonoids induce cell cycle arrest in human prostate cancer cell lines.

    PubMed

    Haddad, A Q; Venkateswaran, V; Viswanathan, L; Teahan, S J; Fleshner, N E; Klotz, L H

    2006-01-01

    Epidemiologic studies have demonstrated an inverse association between flavonoid intake and prostate cancer (PCa) risk. The East Asian diet is very high in flavonoids and, correspondingly, men in China and Japan have the lowest incidence of PCa worldwide. There are thousands of different naturally occurring and synthetic flavonoids. However, only a few have been studied in PCa. Our aim was to identify novel flavonoids with antiproliferative effect in PCa cell lines, as well as determine their effects on cell cycle. We have screened a representative subgroup of 26 flavonoids for antiproliferative effect on the human PCa (LNCaP and PC3), breast cancer (MCF-7), and normal prostate stromal cell lines (PrSC). Using a fluorescence-based cell proliferation assay (Cyquant), we have identified five flavonoids, including the novel compounds 2,2'-dihydroxychalcone and fisetin, with antiproliferative and cell cycle arresting properties in human PCa in vitro. Most of the flavonoids tested exerted antiproliferative effect at lower doses in the PCa cell lines compared to the non-PCa cells. Flow cytometry was used as a means to determine the effects on cell cycle. PC3 cells were arrested in G2/M phase by flavonoids. LNCaP cells demonstrated different cell cycle profiles. Further studies are warranted to determine the molecular mechanism of action of 2,2'-DHC and fisetin in PCa, and to establish their effectiveness in vivo.

  10. Larvicidal and ovideterrent properties of neem oil and fractions against the filariasis vector Aedes albopictus (Diptera: Culicidae): a bioactivity survey across production sites.

    PubMed

    Benelli, Giovanni; Bedini, Stefano; Cosci, Francesca; Toniolo, Chiara; Conti, Barbara; Nicoletti, Marcello

    2015-01-01

    Neem seed oil (NSO) of Azadirachta indica (Meliaceae) contains more than 100 determined biologically active compounds, and many formulations deriving from them showed toxicity, antifeedancy and repellence against a number of arthropod pests. However, it is widely known that botanical products can differ in their chemical composition and bioactivity, as function of the production site and production process. We used high-performance thin layer chromatography (HPTLC) to investigate differences in chemical constituents of NSOs from three production sites. HPTLC analyses showed several differences in chemical abundance and diversity among NSOs, with special reference to limonoids. Furthermore, the three NSOs and their fractions of increasing polarities [i.e. ethyl acetate (EA) fraction and butanol (BU) fraction] were evaluated for larvicidal toxicity and field oviposition deterrence against the Asian tiger mosquito, Aedes albopictus, currently the most invasive mosquito worldwide. Results from bioactivity experiments showed good toxicity of NSOs and EA fractions against A. albopictus fourth instar larvae (with LC50 values ranging from 142.28 to 209.73 ppm), while little toxicity was exerted by BU fractions. A significant effect of the production site and dosage was also found and is probably linked to differences in abundance of constituents among samples, as highlighted by HPTLC analyses. NSOs and EAs were also able to deter A. albopictus oviposition in the field (effective repellence values ranging from 98.55 to 70.10%), while little effectiveness of BU fractions was found. Concerning ovideterrent activity, no difference due to the production site was found. This is the first report concerning larvicidal toxicity of NSO against A. albopictus and ovideterrence against Culicidae in the field. The chance to use chemicals from the NSO EA fraction seems promising, since they are effective at lower doses, if compared to synthetic products currently marketed, and could be

  11. Antiproliferative activity of flavonoids on several cancer cell lines.

    PubMed

    Kawaii, S; Tomono, Y; Katase, E; Ogawa, K; Yano, M

    1999-05-01

    Twenty-seven Citrus flavonoids were examined for their antiproliferative activities against several tumor and normal human cell lines. As a result, 7 flavonoids were judged to be active against the tumor cell lines, while they had weak antiproliferative activity against the normal human cell lines. The rank order of potency was luteolin, natsudaidain, quercetin, tangeretin, eriodictyol, nobiletin, and 3,3',4',5,6,7,8-heptamethoxyflavone. The structure-activity relationship established from comparison among these flavones and flavanones showed that the ortho-catechol moiety in ring B and a C2-C3 double bond were important for the antiproliferative activity. As to polymethoxylated flavones, C-3 hydroxyl and C-8 methoxyl groups were essential for high activity.

  12. Prostaglandin Actions in Established Insect Cell Lines

    USDA-ARS?s Scientific Manuscript database

    Prostaglandins (PGs) are oxygenated metabolites of arachidonic acid (AA) and two other C20 polyunsaturated fatty acids that serve as biochemical signals that mediate a wide range of physiological functions in animal cells. For example, PGs influence protein expression in establish insect cell lines ...

  13. Immune reactivity to dengue and Aedes albopictus mosquitoes in the population from Macao, China, before dengue occurrence.

    PubMed

    De Carvalho, Isabel Lopes; Rocha, Diara Kady; Almeida, A Paulo G

    2011-01-01

    A serological survey was conducted in Macao, China, in 753 individuals, with the objective of looking for antibodies to the mosquito, Aedes albopictus (Skuse 1894) (Diptera: Culicidae), and to dengue, before the occurrence of any autochthonous dengue cases. Blood samples were collected at several public health services, a questionnaire was answered, and enzyme-linked immunosorbant assay (ELISA) and Western blot techniques were performed with extracts of mosquito head and thorax (HT). Anti-Aedes albopictus IgG antibodies were present in titres 1:10(2)-1:10(3) in 9%, and in titres 1:10(4)-1:10(5) in 42% of the sera tested. This reactivity was more frequent (59%) in the population which had resided only in Macao in the 2 years previous to the survey, as opposed to those that had also resided in other areas (50%). From the 230 reactive sera selected for immunoblot, 48 (21%) reacted with a wide range of proteins from above 224 kDa to 21 kDa, with different patterns according to individual sera. Proteins in the intervals 35.3-28.7 kDa and 28.7-21.1 kDa were labelled by the greatest number of sera, 15 and 19 respectively. The presence of anti-Aedes albopictus antibodies presented a statistical relation to skin reaction to mosquito bites, but immunoblot patterns did not. Anti-dengue IgG antibodies were found in 48% of the subjects, with a higher proportion in people who had resided out of Macao, or who were nationals from dengue-endemic neighboring countries. Anti-dengue reactivity was in agreement with anti-mosquito reactivity in half of the population. It would be interesting to see if this proportion has changed since dengue became endemic in Macao in 2001.

  14. Repellent and Larvicidal Activity of the Essential Oil From Eucalyptus nitens Against Aedes aegypti and Aedes albopictus (Diptera: Culicidae).

    PubMed

    Alvarez Costa, Agustín; Naspi, Cecilia V; Lucia, Alejandro; Masuh, Héctor M

    2017-05-01

    Dengue, chikungunya, and yellow fever are important vector-borne diseases transmitted by female mosquitoes when they feed on humans. The use of repellents based on natural products is an alternative for personal protection against these diseases. Application of chemicals with larvicidal activity is another strategy for controlling the mosquito population. The repellent and larvicidal activities of the essential oil from Eucalyptus nitens were tested against Aedes aegypti and Aedes albopictus, the main vectors of these arboviruses. The essential oil was extracted by hydrodistillation and then analyzed by gas chromatography-mass spectrometry. The main components of Eucalyptus nitens essential oil were found to be terpenes such as 1,8-cineole and p-cymene, followed by β-triketones and alkyl esters. The repellent activity of the essential oil against both species was significantly higher when compared with the main component, 1,8-cineole, alone. These results indicate that the repellent effect of E. nitens is not due only to the main component, 1,8-cineole, but also that other compounds may be responsible. Aedes aegypti was found to be more tolerant to the essential oil larvicidal effects than Ae. albopictus (Ae. aegypti LC50 = 52.83 ppm, Ae. albopictus LC 50 = 28.19 ppm). The repellent and larvicidal activity could be associated to the presence of cyclic β-triketones such as flavesone, leptospermone, and isoleptospermone. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  15. Snail regulates cell survival and inhibits cellular senescence in human metastatic prostate cancer cell lines.

    PubMed

    Emadi Baygi, Modjtaba; Soheili, Zahra Soheila; Schmitz, Ingo; Sameie, Shahram; Schulz, Wolfgang A

    2010-12-01

    The epithelial-mesenchymal transition (EMT) is regarded as an important step in cancer metastasis. Snail, a master regulator of EMT, has been recently proposed to act additionally as a cell survival factor and inducer of motility. We have investigated the function of Snail (SNAI1) in prostate cancer cells by downregulating its expression via short (21-mer) interfering RNA (siRNA) and measuring the consequences on EMT markers, cell viability, death, cell cycle, senescence, attachment, and invasivity. Of eight carcinoma cell lines, the prostate carcinoma cell lines LNCaP and PC-3 showed the highest and moderate expression of SNAI1 mRNA, respectively, as measured by quantitative RT-PCR. Long-term knockdown of Snail induced a severe decline in cell numbers in LNCaP and PC-3 and caspase activity was accordingly enhanced in both cell lines. In addition, suppression of Snail expression induced senescence in LNCaP cells. SNAI1-siRNA-treated cells did not tolerate detachment from the extracellular matrix, probably due to downregulation of integrin α6. Expression of E-cadherin, vimentin, and fibronectin was also affected. Invasiveness of PC-3 cells was not significantly diminished by Snail knockdown. Our data suggest that Snail acts primarily as a survival factor and inhibitor of cellular senescence in prostate cancer cell lines. We therefore propose that Snail can act as early driver of prostate cancer progression.

  16. Fucoidan Does Not Exert Anti-Tumorigenic Effects on Uveal Melanoma Cell Lines

    PubMed Central

    Dithmer, Michaela; Kirsch, Anna-Maria; Richert, Elisabeth; Fuchs, Sabine; Wang, Fanlu; Schmidt, Harald; Coupland, Sarah E.; Roider, Johann; Klettner, Alexa

    2017-01-01

    Background. The polysaccharide fucoidan is widely investigated as an anti-cancer agent. Here, we tested the effect of fucoidan on uveal melanoma cell lines. Methods. The effect of 100 µM fucoidan was investigated on five cell lines (92.1, Mel270 OMM1, OMM2.3, OMM2.5) and of 1 µg/mL–1 mg/mL fucoidan in two cell lines (OMM1, OMM2.3). Cell proliferation and viability were investigated with a WST-1 assay, migration in a wound healing (scratch) assay. Vascular Endothelial Growth Factor (VEGF) was measured in ELISA. Angiogenesis was evaluated in co-cultures with endothelial cells. Cell toxicity was induced by hydrogen-peroxide. Protein expression (Akt, ERK1/2, Bcl-2, Bax) was investigated in Western blot. Results. Fucoidan increased proliferation in two and reduced it in one cell line. Migration was reduced in three cell lines. The effect of fucoidan on VEGF was cell type and concentration dependent. In endothelial co-culture with 92.1, fucoidan significantly increased tubular structures. Moreover, fucoidan significantly protected all tested uveal melanoma cell lines from hydrogen-peroxide induced cell death. Under oxidative stress, fucoidan did not alter the expression of Bcl-2, Bax or ERK1/2, while inducing Akt expression in 92.1 cells but not in any other cell line. Conclusion. Fucoidan did not show anti-tumorigenic effects but displayed protective and pro-angiogenic properties, rendering fucoidan unsuitable as a potential new drug for the treatment of uveal melanoma. PMID:28640204

  17. Biosynthesis, characterization, and acute toxicity of Berberis tinctoria-fabricated silver nanoparticles against the Asian tiger mosquito, Aedes albopictus, and the mosquito predators Toxorhynchites splendens and Mesocyclops thermocyclopoides.

    PubMed

    Kumar, Palanisamy Mahesh; Murugan, Kadarkarai; Madhiyazhagan, Pari; Kovendan, Kalimuthu; Amerasan, Duraisamy; Chandramohan, Balamurugan; Dinesh, Devakumar; Suresh, Udaiyan; Nicoletti, Marcello; Alsalhi, Mohamad Saleh; Devanesan, Sandhanasamy; Wei, Hui; Kalimuthu, Kandasamy; Hwang, Jiang-Shiou; Lo Iacono, Annalisa; Benelli, Giovanni

    2016-02-01

    Aedes albopictus is an important arbovirus vector, including dengue. Currently, there is no specific treatment for dengue. Its prevention solely depends on effective vector control measures. In this study, silver nanoparticles (AgNPs) were biosynthesized using a cheap leaf extract of Berberis tinctoria as reducing and stabilizing agent and tested against Ae. albopictus and two mosquito natural enemies. AgNPs were characterized by using UV–vis spectrophotometry, X-ray diffraction, and scanning electron microscopy. In laboratory conditions, the toxicity of AgNPs was evaluated on larvae and pupae of Ae. albopictus. Suitability Index/Predator Safety Factor was assessed on Toxorhynchites splendens and Mesocyclops thermocyclopoides. The leaf extract of B. tinctoria was toxic against larval instars (I–IV) and pupae of Ae. albopictus; LC50 was 182.72 ppm (I instar), 230.99 ppm (II), 269.65 ppm (III), 321.75 ppm (IV), and 359.71 ppm (pupa). B. tinctoria-synthesized AgNPs were highly effective, with LC50 of 4.97 ppm (I instar), 5.97 ppm (II), 7.60 ppm (III), 9.65 ppm (IV), and 14.87 ppm (pupa). Both the leaf extract and AgNPs showed reduced toxicity against the mosquito natural enemies M. thermocyclopoides and T. splendens. Overall, this study firstly shed light on effectiveness of B. tinctoria-synthesized AgNPs as an eco-friendly nanopesticide, highlighting the concrete possibility to employ this newer and safer tool in arbovirus vector control programs.

  18. Combining the Sterile Insect Technique with the Incompatible Insect Technique: I-Impact of Wolbachia Infection on the Fitness of Triple- and Double-Infected Strains of Aedes albopictus

    PubMed Central

    Zhang, Dongjing; Zheng, Xiaoying; Xi, Zhiyong; Bourtzis, Kostas; Gilles, Jeremie R. L.

    2015-01-01

    The mosquito species Aedes albopictus is a major vector of the human diseases dengue and chikungunya. Due to the lack of efficient and sustainable methods to control this mosquito species, there is an increasing interest in developing and applying the sterile insect technique (SIT) and the incompatible insect technique (IIT), separately or in combination, as population suppression approaches. Ae. albopictus is naturally double-infected with two Wolbachia strains, wAlbA and wAlbB. A new triple Wolbachia-infected strain (i.e., a strain infected with wAlbA, wAlbB, and wPip), known as HC and expressing strong cytoplasmic incompatibility (CI) in appropriate matings, was recently developed. In the present study, we compared several fitness traits of three Ae. albopictus strains (triple-infected, double-infected and uninfected), all of which were of the same genetic background (“Guangzhou City, China”) and were reared under the same conditions. Investigation of egg-hatching rate, survival of pupae and adults, sex ratio, duration of larval stages (development time from L1 to pupation), time to emergence (development time from L1 to adult emergence), wing length, female fecundity and adult longevity indicated that the presence of Wolbachia had only a minimal effect on host fitness. Based on this evidence, the HC strain is currently under consideration for mass rearing and application in a combined SIT-IIT strategy to control natural populations of Ae. albopictus in mainland China. PMID:25849812

  19. Evaluation of procoagulant tissue factor expression in canine hemangiosarcoma cell lines.

    PubMed

    Witter, Lauren E; Gruber, Erika J; Lean, Fabian Z X; Stokol, Tracy

    2017-01-01

    OBJECTIVE To evaluate expression of procoagulant tissue factor (TF) by canine hemangiosarcoma cells in vitro. SAMPLES 4 canine hemangiosarcoma cell lines (SB-HSA [mouse-passaged cutaneous tumor], Emma [primary metastatic brain tumor], and Frog and Dal-1 [primary splenic tumors]) and 1 nonneoplastic canine endothelial cell line (CnAoEC). PROCEDURES TF mRNA and TF antigen expression were evaluated by quantitative real-time PCR assay and flow cytometry, respectively. Thrombin generation was measured in canine plasma and in coagulation factor-replete or specific coagulation factor-deficient human plasma by calibrated automated thrombography. Corn trypsin inhibitor and annexin V were used to examine contributions of contact activation and membrane-bound phosphatidylserine, respectively, to thrombin generation. RESULTS All cell lines expressed TF mRNA and antigen, with significantly greater expression of both products in SB-HSA and Emma cells than in CnAoEC. A greater percentage of SB-HSA cells expressed TF antigen, compared with other hemangiosarcoma cell lines. All hemangiosarcoma cell lines generated significantly more thrombin than did CnAoEC in canine or factor-replete human plasma. Thrombin generation induced by SB-HSA cells was significantly lower in factor VII-deficient plasma than in factor-replete plasma and was abolished in factor X-deficient plasma; residual thrombin generation in factor VII-deficient plasma was abolished by incubation of cells with annexin V. Thrombin generation by SB-HSA cells was unaffected by the addition of corn trypsin inhibitor. CONCLUSIONS AND CLINICAL RELEVANCE Hemangiosarcoma cell lines expressed procoagulant TF in vitro. Further research is needed to determine whether TF can be used as a biomarker for hemostatic dysfunction in dogs with hemangiosarcoma.

  20. Evaluation of procoagulant tissue factor expression in canine hemangiosarcoma cell lines

    PubMed Central

    Witter, Lauren E.; Gruber, Erika J.; Lean, Fabian Z. X.; Stokol, Tracy

    2017-01-01

    OBJECTIVE To evaluate expression of procoagulant tissue factor (TF) by canine hemangiosarcoma cells in vitro. SAMPLES 4 canine hemangiosarcoma cell lines (SB-HSA [mouse-passaged cutaneous tumor], Emma [primary metastatic brain tumor], and Frog and Dal-1 [primary splenic tumors]) and 1 nonneoplastic canine endothelial cell line (CnAoEC). PROCEDURES TF mRNA and TF antigen expression were evaluated by quantitative real-time PCR assay and flow cytometry, respectively. Thrombin generation was measured in canine plasma and in coagulation factor–replete or specific coagulation factor–deficient human plasma by calibrated automated thrombography. Corn trypsin inhibitor and annexin V were used to examine contributions of contact activation and membrane-bound phosphatidylserine, respectively, to thrombin generation. RESULTS All cell lines expressed TF mRNA and antigen, with significantly greater expression of both products in SB-HSA and Emma cells than in CnAoEC. A greater percentage of SB-HSA cells expressed TF antigen, compared with other hemangiosarcoma cell lines. All hemangiosarcoma cell lines generated significantly more thrombin than did CnAoEC in canine or factor-replete human plasma. Thrombin generation induced by SB-HSA cells was significantly lower in factor VII-deficient plasma than in factor-replete plasma and was abolished in factor X–deficient plasma; residual thrombin generation in FVII-deficient plasma was abolished by incubation of cells with annexin V. Thrombin generation by SB-HSA cells was unaffected by the addition of corn trypsin inhibitor. CONCLUSIONS AND CLINICAL RELEVANCE Hemangiosarcoma cell lines expressed procoagulant TF in vitro. Further research is needed to determine whether TF can be used as a biomarker for hemostatic dysfunction in dogs with hemangiosarcoma. PMID:28029283

  1. Modelling the breeding of Aedes Albopictus species in an urban area in Pulau Pinang using polynomial regression

    NASA Astrophysics Data System (ADS)

    Salleh, Nur Hanim Mohd; Ali, Zalila; Noor, Norlida Mohd.; Baharum, Adam; Saad, Ahmad Ramli; Sulaiman, Husna Mahirah; Ahmad, Wan Muhamad Amir W.

    2014-07-01

    Polynomial regression is used to model a curvilinear relationship between a response variable and one or more predictor variables. It is a form of a least squares linear regression model that predicts a single response variable by decomposing the predictor variables into an nth order polynomial. In a curvilinear relationship, each curve has a number of extreme points equal to the highest order term in the polynomial. A quadratic model will have either a single maximum or minimum, whereas a cubic model has both a relative maximum and a minimum. This study used quadratic modeling techniques to analyze the effects of environmental factors: temperature, relative humidity, and rainfall distribution on the breeding of Aedes albopictus, a type of Aedes mosquito. Data were collected at an urban area in south-west Penang from September 2010 until January 2011. The results indicated that the breeding of Aedes albopictus in the urban area is influenced by all three environmental characteristics. The number of mosquito eggs is estimated to reach a maximum value at a medium temperature, a medium relative humidity and a high rainfall distribution.

  2. In vitro effects of Apixaban on 5 different cancer cell lines

    PubMed Central

    Guasti, Luigina; Moretto, Paola; Vigetti, Davide; Ageno, Walter; Dentali, Francesco; Maresca, Andrea M.; Campiotti, Leonardo; Grandi, Anna M.; Passi, Alberto

    2017-01-01

    Background Cancer is associated with hypercoagulability. However, several data suggest that anticoagulant drugs may have an effect on tumor development and progression mediated by both coagulation dependent processes and non-coagulation dependent processes. Therefore, we investigated the in vitro effects of Apixaban on cell proliferation, mortality, cell migration, gene expression and matrix metalloproteinase in 5 different cancer cell lines. Methods The following cancer cell lines, and 2 normal fibroblast cultures (lung and dermal fibroblasts), were studied: OVCAR3 (ovarian cancer), MDA MB 231 (breast cancer), CaCO-2 (colon cancer), LNCaP (prostate cancer) and U937 (histiocytic lymphoma). Proliferation and cell mortality were assessed in control cells and Apixaban treated cultures (dose from 0.1 to 5 μg/ml, 0 to 96-h). Necrosis/Apoptosis (fluorescence microscopy), cell migration (24-h after scratch test), matrix metalloproteinase (MMP) activity and mRNA expression (RT PCR) of p16, p21, p53 and HAS were also assessed. Results High-dose (5 μg/ml) Apixaban incubation was associated with a significantly reduced proliferation in 3 cancer cell lines (OVCAR3, CaCO-2 and LNCaP) and with increased cancer cell mortality in all, except LNCaP, cancer lines. Apoptosis seems to account for the increased mortality. The migration capacity seems to be impaired after high-dose Apixaban incubation in OVCAR3 and CaCO-2 cells. Data on mRNA expression suggest a consistent increase in tumor suppression gene p16 in all cell lines. Conclusions Our data suggest that high-dose Apixaban may be able to interfere with cancer cell in vitro, reducing proliferation and increasing cancer cell mortality through apoptosis in several cancer cell lines. PMID:29023465

  3. Metabolic Response to NAD Depletion across Cell Lines Is Highly Variable.

    PubMed

    Xiao, Yang; Kwong, Mandy; Daemen, Anneleen; Belvin, Marcia; Liang, Xiaorong; Hatzivassiliou, Georgia; O'Brien, Thomas

    2016-01-01

    Nicotinamide adenine dinucleotide (NAD) is a cofactor involved in a wide range of cellular metabolic processes and is a key metabolite required for tumor growth. NAMPT, nicotinamide phosphoribosyltransferase, which converts nicotinamide (NAM) to nicotinamide mononucleotide (NMN), the immediate precursor of NAD, is an attractive therapeutic target as inhibition of NAMPT reduces cellular NAD levels and inhibits tumor growth in vivo. However, there is limited understanding of the metabolic response to NAD depletion across cancer cell lines and whether all cell lines respond in a uniform manner. To explore this we selected two non-small cell lung carcinoma cell lines that are sensitive to the NAMPT inhibitor GNE-617 (A549, NCI-H1334), one that shows intermediate sensitivity (NCI-H441), and one that is insensitive (LC-KJ). Even though NAD was reduced in all cell lines there was surprising heterogeneity in their metabolic response. Both sensitive cell lines reduced glycolysis and levels of di- and tri-nucleotides and modestly increased oxidative phosphorylation, but they differed in their ability to combat oxidative stress. H1334 cells activated the stress kinase AMPK, whereas A549 cells were unable to activate AMPK as they contain a mutation in LKB1, which prevents activation of AMPK. However, A549 cells increased utilization of the Pentose Phosphate pathway (PPP) and had lower reactive oxygen species (ROS) levels than H1334 cells, indicating that A549 cells are better able to modulate an increase in oxidative stress. Inherent resistance of LC-KJ cells is associated with higher baseline levels of NADPH and a delayed reduction of NAD upon NAMPT inhibition. Our data reveals that cell lines show heterogeneous response to NAD depletion and that the underlying molecular and genetic framework in cells can influence the metabolic response to NAMPT inhibition.

  4. Establishment of cell lines from adult T-cell leukemia cells dependent on negatively charged polymers.

    PubMed

    Kagami, Yoshitoyo; Uchiyama, Susumu; Kato, Harumi; Okada, Yasutaka; Seto, Masao; Kinoshita, Tomohiro

    2017-07-05

    Growing adult T-cell leukemia/lymphoma (ATLL) cells in vitro is difficult. Here, we examined the effects of static electricity in the culture medium on the proliferation of ATLL cells. Six out of 10 ATLL cells did not proliferate in vitro and thus had to be cultured in a medium containing negatively charged polymers. In the presence of poly-γ-glutamic acid (PGA) or chondroitin sulfate (CDR), cell lines (HKOX3-PGA, HKOX3-CDR) were established from the same single ATLL case using interleukin (IL)-2, IL-4, and feeder cells expressing OX40L (OX40L + HK). Dextran sulfate inhibited growth in both HKOX3 cell lines. Both PGA and OX40L + HK were indispensable for HKOX3-PGA growth, but HKOX3-CDR could proliferate in the presence of CDR or OX40L + HK alone. Thus, the specific action of each negatively charged polymer promoted the growth of specific ATLL cells in vitro.

  5. Lessons From the First Comprehensive Molecular Characterization of Cell Cycle Control in Rodent Insulinoma Cell Lines

    PubMed Central

    Cozar-Castellano, Irene; Harb, George; Selk, Karen; Takane, Karen; Vasavada, Rupangi; Sicari, Brian; Law, Brian; Zhang, Pili; Scott, Donald K.; Fiaschi-Taesch, Nathalie; Stewart, Andrew F.

    2008-01-01

    OBJECTIVE—Rodent insulinoma cell lines may serve as a model for designing continuously replicating human β-cell lines and provide clues as to the central cell cycle regulatory molecules in the β-cell. RESEARCH DESIGN AND METHODS—We performed a comprehensive G1/S proteome analysis on the four most widely studied rodent insulinoma cell lines and defined their flow cytometric profiles and growth characteristics. RESULTS—1) Despite their common T-antigen–derived origins, MIN6 and BTC3 cells display markedly different G1/S expression profiles; 2) despite their common radiation origins, RINm5F and INS1 cells display striking differences in cell cycle protein profiles; 3) phosphorylation of pRb is absent in INS1 and RINm5F cells; 4) cyclin D2 is absent in RINm5F and BTC3 cells and therefore apparently dispensable for their proliferation; 5) every cell cycle inhibitor is upregulated, presumably in a futile attempt to halt proliferation; 6) among the G1/S proteome members, seven are pro-proliferation molecules: cyclin-dependent kinase-1, -2, -4, and -6 and cyclins A, E, and D3; and 7) overexpression of the combination of these seven converts arrested proliferation rates in primary rat β-cells to those in insulinoma cells. Unfortunately, this therapeutic overexpression appears to mildly attenuate β-cell differentiation and function. CONCLUSIONS—These studies underscore the importance of characterizing the cell cycle at the protein level in rodent insulinoma cell lines. They also emphasize the hazards of interpreting data from rodent insulinoma cell lines as modeling normal cell cycle progression. Most importantly, they provide seven candidate targets for inducing proliferation in human β-cells. PMID:18650366

  6. Characterization of Tolypocladium cylindrosporum (Hypocreales: Ophiocordycipitacea) and its impact against Aedes aegypti and Aedes albopictus eggs at low temperature

    USDA-ARS?s Scientific Manuscript database

    We examined the growth characteristics of Tolypocladium cylindrosporum IBT 41712 and its potential to infect eggs of Aedes aegypti Linnaeus and Aedes albopictus (Skuse) mosquitoes at low temperature (15 deg C). When grown on sabouraud dextrose agar supplemented with yeast extract, IBT 41712 formed w...

  7. Nanotopography induced contact guidance of the F11 cell line during neuronal differentiation: a neuronal model cell line for tissue scaffold development

    NASA Astrophysics Data System (ADS)

    Wieringa, Paul; Tonazzini, Ilaria; Micera, Silvestro; Cecchini, Marco

    2012-07-01

    The F11 hybridoma, a dorsal root ganglion-derived cell line, was used to investigate the response of nociceptive sensory neurons to nanotopographical guidance cues. This established this cell line as a model of peripheral sensory neuron growth for tissue scaffold design. Cells were seeded on substrates of cyclic olefin copolymer (COC) films imprinted via nanoimprint lithography (NIL) with a grating pattern of nano-scale grooves and ridges. Different ridge widths were employed to alter the focal adhesion formation, thereby changing the cell/substrate interaction. Differentiation was stimulated with forskolin in culture medium consisting of either 1 or 10% fetal bovine serum (FBS). Per medium condition, similar neurite alignment was achieved over the four day period, with the 1% serum condition exhibiting longer, more aligned neurites. Immunostaining for focal adhesions found the 1% FBS condition to also have fewer, less developed focal adhesions. The robust response of the F11 to guidance cues further builds on the utility of this cell line as a sensory neuron model, representing a useful tool to explore the design of regenerative guidance tissue scaffolds.

  8. Culture of human cell lines by a pathogen-inactivated human platelet lysate.

    PubMed

    Fazzina, R; Iudicone, P; Mariotti, A; Fioravanti, D; Procoli, A; Cicchetti, E; Scambia, G; Bonanno, G; Pierelli, L

    2016-08-01

    Alternatives to the use of fetal bovine serum (FBS) have been investigated to ensure xeno-free growth condition. In this study we evaluated the efficacy of human platelet lysate (PL) as a substitute of FBS for the in vitro culture of some human cell lines. PL was obtained by pools of pathogen inactivated human donor platelet (PLT) concentrates. Human leukemia cell lines (KG-1, K562, JURKAT, HL-60) and epithelial tumor cell lines (HeLa and MCF-7) were cultured with either FBS or PL. Changes in cell proliferation, viability, morphology, surface markers and cell cycle were evaluated for each cell line. Functional characteristics were analysed by drug sensitivity test and cytotoxicity assay. Our results demonstrated that PL can support growth and expansion of all cell lines, although the cells cultured in presence of PL experienced a less massive proliferation compared to those grown with FBS. We found a comparable percentage of viable specific marker-expressing cells in both conditions, confirming lineage fidelity in all cultures. Functionality assays showed that cells in both FBS- and PL-supported cultures maintained their normal responsiveness to adriamycin and NK cell-mediated lysis. Our findings indicate that PL is a feasible serum substitute for supporting growth and propagation of haematopoietic and epithelial cell lines with many advantages from a perspective of process standardization, ethicality and product safety.

  9. Isolation, Characterization, and Establishment of Spontaneously Immortalized Cell Line HRPE-2S With Stem Cell Properties.

    PubMed

    Shams Najafabadi, Hoda; Soheili, Zahra-Soheila; Samiei, Shahram; Ahmadieh, Hamid; Ranaei Pirmardan, Ehsan; Masoumi, Maryam

    2017-10-01

    The retinal pigment epithelium is a monolayer of highly specialized pigmented cells located between the neural retina and the Bruch's membrane of the choroid. RPE cells play a crucial role in the maintenance and function of the underlying photoreceptors. This study introduces a spontaneously arising human retinal pigment epithelial cell line, HRPE-2S, which was isolated from primary RPE cell culture of 2 days old male donor. We characterized morphology and functional properties of the new cell line. The immortalized cell line was maintained in culture for more than 70 passages and 240 divisions. The average doubling time of the cells was approximately 22 h and got freezed at 26th passage. The cell line expressed RPE-specific markers RPE65 and cell junction protein ZO1 as an epithelial cell marker. It also expressed CHX10, PAX6, Nestin, SOX2 as stem and retinal progenitor cell markers. Ki67 as a marker of cell proliferation was expressed in all HRPE-2S cells. It represented typical epithelial cobblestone morphology and did not phenotypically change through several passages. Stem cell-like aggregations (neurospheres) were observed in SEM microscopy. The cells represented high mitotic index. They could be viable under hypoxic conditions and serum deprivation. According to functional studies, the cell line exhibited stem cell-like behaviors with particular emphasis on its self-renewal capacity. LDH isoenzymes expression pattern confirmed the same cellular source for both of the HRPE-2S cells and primary RPE cells. Characteristics of HRPE-2S cells promise it as an in vitro model for RPE stem cell-based researches. J. Cell. Physiol. 232: 2626-2640, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  10. Immortal hepatic stellate cell lines: useful tools to study hepatic stellate cell biology and function?

    PubMed Central

    Herrmann, Jens; Gressner, Axel M; Weiskirchen, Ralf

    2007-01-01

    Abstract At the cellular level, the activation and transdifferentiation of quiescent hepatic stellate cells (HSC) into myofibroblasts is the key process involved in hepatic fibrogenesis that is associated with an increased and altered deposition of extracellular matrix components in the liver. The temporal sequence of molecular events associated with stellate cell activation turned out to be appropriately mimicked when HSC isolated from normal livers are cultured on uncoated plastic surface. Therefore, cultured primary cells isolated from rodents and human beings are common in vitro models in investigations addressing these issues of hepatic stellate biology and function. However, the limited supply, cost-effective isolation procedure and the ever growing need have resulted in efforts to establish immortalized stellate cell lines having the advantage of virtually unlimited access. They allow rapid screening for disease-associated factors and restrict the necessary number of animal experiments. From the first description of an immortal HSC line in 1986, a huge number of studies were conducted with these established cell lines. However, differences in morphology, growth characteristics and anomalies of chromosome number and structure make the applications of these models questionable. Here, we summarize the history and cellular characteristics of respective cell lines and discuss the differences of continuous HSC lines and their primary counterparts. PMID:17760834

  11. Immortal hepatic stellate cell lines: useful tools to study hepatic stellate cell biology and function?

    PubMed

    Herrmann, Jens; Gressner, Axel M; Weiskirchen, Ralf

    2007-01-01

    At the cellular level, the activation and transdifferentiation of quiescent hepatic stellate cells (HSC) into myofibroblasts is the key process involved in hepatic fibrogenesis that is associated with an increased and altered deposition of extracellular matrix components in the liver. The temporal sequence of molecular events associated with stellate cell activation turned out to be appropriately mimicked when HSC isolated from normal livers are cultured on uncoated plastic surface. Therefore, cultured primary cells isolated from rodents and human beings are common in vitro models in investigations addressing these issues of hepatic stellate biology and function. However, the limited supply, cost-effective isolation procedure and the ever growing need have resulted in efforts to establish immortalized stellate cell lines having the advantage of virtually unlimited access. They allow rapid screening for disease-associated factors and restrict the necessary number of animal experiments. From the first description of an immortal HSC line in 1986, a huge number of studies were conducted with these established cell lines. However, differences in morphology, growth characteristics and anomalies of chromosome number and structure make the applications of these models questionable. Here, we summarize the history and cellular characteristics of respective cell lines and discuss the differences of continuous HSC lines and their primary counterparts.

  12. Isolation and characterization of conditionally immortalized mouse glomerular endothelial cell lines.

    PubMed

    Rops, Angelique L; van der Vlag, Johan; Jacobs, Cor W; Dijkman, Henry B; Lensen, Joost F; Wijnhoven, Tessa J; van den Heuvel, Lambert P; van Kuppevelt, Toin H; Berden, Jo H

    2004-12-01

    The culture and establishment of glomerular cell lines has proven to be an important tool for the understanding of glomerular cell functions in glomerular physiology and pathology. Especially, the recent establishment of a conditionally immortalized visceral epithelial cell line has greatly boosted the research on podocyte biology. Glomeruli were isolated from H-2Kb-tsA58 transgenic mice that contain a gene encoding a temperature-sensitive variant of the SV40 large tumor antigen, facilitating proliferative growth at 33 degrees C and differentiation at 37 degrees C. Glomerular endothelial cells were isolated from glomerular outgrowth by magnetic beads loaded with CD31, CD105, GSL I-B4, and ULEX. Clonal cell lines were characterized by immunofluorescence staining with antibodies/lectins specific for markers of endothelial cells, podocytes, and mesangial cells. Putative glomerular endothelial cell lines were analyzed for (1) cytokine-induced expression of adhesion molecules; (2) tube formation on Matrigel coating; and (3) the presence of fenestrae. As judged by immunostaining for Wilms tumor-1, smooth muscle actin (SMA), podocalyxin, and von Willebrand factor (vWF), we obtained putative endothelial, podocyte and mesangial cell lines. The mouse glomerular endothelial cell clone #1 (mGEnC-1) was positive for vWF, podocalyxin, CD31, CD105, VE-cadherin, GSL I-B4, and ULEX, internalized acetylated-low-density lipoprotein (LDL), and showed increased expression of adhesion molecules after activation with proinflammatory cytokines. Furthermore, mGEnC-1 formed tubes and contained nondiaphragmed fenestrae. The mGEnC-1 represents a conditionally immortalized cell line with various characteristics of differentiated glomerular endothelial cells when cultured at 37 degrees C. Most important, mGEnC-1 contains nondiaphragmed fenestrae, which is a unique feature of glomerular endothelial cells.

  13. Parallel RNAi screens across different cell lines identify generic and cell type-specific regulators of actin organization and cell morphology.

    PubMed

    Liu, Tao; Sims, David; Baum, Buzz

    2009-01-01

    In recent years RNAi screening has proven a powerful tool for dissecting gene functions in animal cells in culture. However, to date, most RNAi screens have been performed in a single cell line, and results then extrapolated across cell types and systems. Here, to dissect generic and cell type-specific mechanisms underlying cell morphology, we have performed identical kinome RNAi screens in six different Drosophila cell lines, derived from two distinct tissues of origin. This analysis identified a core set of kinases required for normal cell morphology in all lines tested, together with a number of kinases with cell type-specific functions. Most significantly, the screen identified a role for minibrain (mnb/DYRK1A), a kinase associated with Down's syndrome, in the regulation of actin-based protrusions in CNS-derived cell lines. This cell type-specific requirement was not due to the peculiarities in the morphology of CNS-derived cells and could not be attributed to differences in mnb expression. Instead, it likely reflects differences in gene expression that constitute the cell type-specific functional context in which mnb/DYRK1A acts. Using parallel RNAi screens and gene expression analyses across cell types we have identified generic and cell type-specific regulators of cell morphology, which include mnb/DYRK1A in the regulation of protrusion morphology in CNS-derived cell lines. This analysis reveals the importance of using different cell types to gain a thorough understanding of gene function across the genome and, in the case of kinases, the difficulties of using the differential gene expression to predict function.

  14. Establishment and characterization of fetal fibroblast cell lines for generating human lysozyme transgenic goats by somatic cell nuclear transfer.

    PubMed

    Liu, Jun; Luo, Yan; Zheng, Liming; Liu, Qingqing; Yang, Zhongcai; Wang, Yongsheng; Su, Jianmin; Quan, Fusheng; Zhang, Yong

    2013-10-01

    This study was performed to qualify goat fetal fibroblast (GFF) cell lines for genetic modification and somatic cell nuclear transfer (SCNT) to produce human lysozyme (hLYZ) transgenic goats. Nine GFF cell lines were established from different fetuses, and the proliferative lifespan and chromosomal stability were analyzed. The results suggested that cell lines with a longer lifespan had stable chromosomes compared with those of cells lines with a shorter lifespan. According to the proliferative lifespan, we divided GFF cell lines into two groups: cell lines with a long lifespan (GFF1/2/7/8/9; group L) and cell lines with a short lifespan (GFF3/4/5/6; group S). Next, a hLYZ expression vector was introduced into these cell lines by electroporation. The efficiencies of colony formation, expansion in culture, and the quality of transgenic clonal cell lines were significant higher in group L than those in group S. The mean fusion rate and blastocyst rate in group L were higher than those in group S (80.3 ± 1.7 vs. 65.1 ± 4.2 % and 19.5 ± 0.6 vs. 15.1 ± 1.1 %, respectively, P < 0.05). After transferring cloned embryos into the oviducts of recipient goats, three live kids were born. PCR and Southern blot analyses confirmed integration of the transgene in cloned goats. In conclusion, the lifespan of GFF cell lines has a major effect on the efficiency to produce transgenic cloned goats. Therefore, the proliferative lifespan of primary cells may be used as a criterion to characterize the quality of cell lines for genetic modification and SCNT.

  15. Spontaneous lung metastasis formation of human Merkel cell carcinoma cell lines transplanted into scid mice.

    PubMed

    Knips, Jill; Czech-Sioli, Manja; Spohn, Michael; Heiland, Max; Moll, Ingrid; Grundhoff, Adam; Schumacher, Udo; Fischer, Nicole

    2017-07-01

    Merkel cell carcinoma (MCC) is an aggressive skin cancer entity that frequently leads to rapid death due to its high propensity to metastasize. The etiology of most MCC cases is linked to Merkel cell polyomavirus (MCPyV), a virus which is monoclonally integrated in up to 95% of tumors. While there are presently no animal models to study the role of authentic MCPyV infection on transformation, tumorigenesis or metastasis formation, xenograft mouse models employing engrafted MCC-derived cell lines (MCCL) represent a promising approach to study certain aspects of MCC pathogenesis. Here, the two MCPyV-positive MCC cell lines WaGa and MKL-1 were subcutaneously engrafted in scid mice. Engraftment of both MCC cell lines resulted in the appearance of circulating tumor cells and metastasis formation, with WaGa-engrafted mice showing a significantly shorter survival time as well as increased numbers of spontaneous lung metastases compared to MKL-1 mice. Interestingly, explanted tumors compared to parental cell lines exhibit an upregulation of MCPyV sT-Antigen expression in all tumors, with WaGa tumors showing significantly higher sT-Antigen expression than MKL-1 tumors. RNA-Seq analysis of explanted tumors and parental cell lines furthermore revealed that in the more aggressive WaGa tumors, genes involved in inflammatory response, growth factor activity and Wnt signalling pathway are significantly upregulated, suggesting that sT-Antigen is the driver of the observed differences in metastasis formation. © 2017 UICC.

  16. Developing global regression models for metabolite concentration prediction regardless of cell line.

    PubMed

    André, Silvère; Lagresle, Sylvain; Da Sliva, Anthony; Heimendinger, Pierre; Hannas, Zahia; Calvosa, Éric; Duponchel, Ludovic

    2017-11-01

    Following the Process Analytical Technology (PAT) of the Food and Drug Administration (FDA), drug manufacturers are encouraged to develop innovative techniques in order to monitor and understand their processes in a better way. Within this framework, it has been demonstrated that Raman spectroscopy coupled with chemometric tools allow to predict critical parameters of mammalian cell cultures in-line and in real time. However, the development of robust and predictive regression models clearly requires many batches in order to take into account inter-batch variability and enhance models accuracy. Nevertheless, this heavy procedure has to be repeated for every new line of cell culture involving many resources. This is why we propose in this paper to develop global regression models taking into account different cell lines. Such models are finally transferred to any culture of the cells involved. This article first demonstrates the feasibility of developing regression models, not only for mammalian cell lines (CHO and HeLa cell cultures), but also for insect cell lines (Sf9 cell cultures). Then global regression models are generated, based on CHO cells, HeLa cells, and Sf9 cells. Finally, these models are evaluated considering a fourth cell line(HEK cells). In addition to suitable predictions of glucose and lactate concentration of HEK cell cultures, we expose that by adding a single HEK-cell culture to the calibration set, the predictive ability of the regression models are substantially increased. In this way, we demonstrate that using global models, it is not necessary to consider many cultures of a new cell line in order to obtain accurate models. Biotechnol. Bioeng. 2017;114: 2550-2559. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  17. Pharmacogenomic agreement between two cancer cell line data sets.

    PubMed

    2015-12-03

    Large cancer cell line collections broadly capture the genomic diversity of human cancers and provide valuable insight into anti-cancer drug response. Here we show substantial agreement and biological consilience between drug sensitivity measurements and their associated genomic predictors from two publicly available large-scale pharmacogenomics resources: The Cancer Cell Line Encyclopedia and the Genomics of Drug Sensitivity in Cancer databases.

  18. Field Comparisons of the Gravid Aedes Trap (GAT) and BG-Sentinel Trap for Monitoring Aedes albopictus (Diptera: Culicidae) Populations and Notes on Indoor GAT Collections in Vietnam.

    PubMed

    Johnson, Brian J; Hurst, Tim; Quoc, Hung Luu; Unlu, Isik; Freebairn, Christopher; Faraji, Ary; Ritchie, Scott A

    2017-03-01

    We report on the use of the Gravid Aedes Trap (GAT) as a surveillance device for Aedes albopictus (Skuse) relative to the BG-Sentinel (BGS) trap in field studies conducted in Trenton, NJ, and on Hammond Island, Queensland, Australia. A parallel study conducted in Nha Trang, Vietnam, assessed the use of the GAT as an indoor surveillance device as well as the use of canola oil as a noninsecticide killing agent. In Trenton and Hammond Island, the GAT collected fewer male (0.40 ± 0.12 and 0.43 ± 0.30, respectively) and female (3.05 ± 0.67 and 2.7 ± 2.3, respectively) Ae. albopictus than the BGS trap (males: 3.54 ± 1.26 and 3.75 ± 0.83; females: 4.66 ± 1.18 and 3.9 ± 0.23) over their respective sampling periods (i.e., 24 h for the BGS and 1 wk for the GAT). Despite differences in capture rates, the percentage of traps positive for female Ae. albopictus was similar between the BGS and GAT (Trenton: 60.1 ± 6.3% and 64.4 ± 4.1%; Hammond: 87.5 ± 6.9% and 80.0 ± 8.2%). In Nha Trang, the GAT was equally effective indoors and outdoors with (10 g hay or 3 g fish food) and without (water or empty) infusion. Additionally, no significant decrease in collections was observed between GATs set with canola oil or long-lasting insecticidal net. In summary, both traps were successful in monitoring female Ae. albopictus over their respective trapping intervals, but would be best used to complement each other to monitor both sexes and all physiological stages of female Ae. albopictus. However, the versatility and low-cost of the GAT makes it an attractive alternative to the more expensive BGS trap. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  19. [Establishment of fibroblast cell line and its biological characteristics in Matou goat].

    PubMed

    Li, Tianda; Liu, Chousheng; Wang, Zhigang; Zhang, Liping; Sun, Xiuzhu; Zhao, Junjin; Meng, Fei; Luo, Guihe; Zhu, Jinqing

    2008-12-01

    Taking Matou goat ear margin as the study material, we succeeded in established a fibroblast cell line by the method of explant culture directly. Observations on morphology, dynamic growth, determination of viability, analysis of karyotype, test of microorganism and other characteristics were detected. Results showed: Population Doubling Time (PDT) of cells was approximately 36 h; Cell viability was 96.7% after thawing; The status of cell After passage was constant; Analysis of chromosomal karyotyps indicated that diploid (2n=60) account for 98% in the cell line. Every index in the cell line met all the standard quality controls of ATCC in USA. The established of Matou goat ear fibroblast cell line has not only important genetic resources preserved at the cell level, but also valuable material for genome, postgenome and somatic cell nuclear transfer research.

  20. Derivation and characterization of Chinese human embryonic stem cell line with high potential to differentiate into pancreatic and hepatic cells.

    PubMed

    Shi, Cheng; Shen, Huan; Jiang, Wei; Song, Zhi-Hua; Wang, Cheng-Yan; Wei, Li-Hui

    2011-04-01

    Human embryonic stem cells have prospective uses in regenerative medicine and drug screening. Every human embryonic stem cell line has its own genetic background, which determines its specific ability for differentiation as well as susceptibility to drugs. It is necessary to compile many human embryonic stem cell lines with various backgrounds for future clinical use, especially in China due to its large population. This study contributes to isolating new Chinese human embryonic stem cell lines with clarified directly differentiation ability. Donated embryos that exceeded clinical use in our in vitro fertilization-embryo transfer (IVF-ET) center were collected to establish human embryonic stem cells lines with informed consent. The classic growth factors of basic fibroblast growth factor (bFGF) and recombinant human leukaemia inhibitory factor (hLIF) for culturing embryonic stem cells were used to capture the stem cells from the plated embryos. Mechanical and enzymetic methods were used to propagate the newly established human embryonic stem cells line. The new cell line was checked for pluripotent characteristics with detecting the expression of stemness genes and observing spontaneous differentiation both in vitro and in vivo. Finally similar step-wise protocols from definitive endoderm to target specific cells were used to check the cell line's ability to directly differentiate into pancreatic and hepatic cells. We generated a new Chinese human embryonic stem cells line, CH1. This cell line showed the same characteristics as other reported Chinese human embryonic stem cells lines: normal morphology, karyotype and pluripotency in vitro and in vivo. The CH1 cells could be directly differentiated towards pancreatic and hepatic cells with equal efficiency compared to the H1 cell line. This newly established Chinese cell line, CH1, which is pluripotent and has high potential to differentiate into pancreatic and hepatic cells, will provide a useful tool for embryo

  1. Study of the distribution and abundance of the eggs of Aedes aegypti and Aedes albopictus according to the habitat and meteorological variables, municipality of São Sebastião, São Paulo State, Brazil

    PubMed Central

    2013-01-01

    Background This study focused on the distribution and abundance of the eggs of Aedes aegypti and Aedes albopictus. Methods Eighty ovitraps were exposed for four days of each month in peri- and intradomiciliary environments of 40 urban residences on 20 street blocks that were drawn monthly in Sebastião, SP, between February 2011 and February 2012. The monthly distribution of positive ovitrap indices (POI) and mean egg counts per trap (MET) of Ae. aegypti and Ae. albopictus were analyzed using the Kruskal-Wallis test, followed by the Dwass-Steel-Critchlow-Fligner (DSCF) test. Spearman's rank correlation coefficient and simple linear regression were used to determine the association between the meteorological variables of temperature and rainfall and the number of ovitraps with eggs and the egg count. Results The POI and MET of Ae. aegypti were higher in peridomiciliary premises. A positive correlation was found between the temperature and the number of ovitraps with eggs and the egg count of this species in domestic environments. There was no difference in the POI and MET of Ae. albopictus between the environments. A positive correlation was found between temperature and positive ovitraps of Ae. albopictus in peridomiciliary premises. The POI and MET of Ae. aegypti were higher than those of Ae. albopictus. Conclusions Peridomiciliary premises were the preferred environments for oviposition of Ae. aegypti. The use of ovitraps for surveillance and vector control is reiterated. PMID:24499530

  2. RUNX3 confers sensitivity to pheophorbide a-photodynamic therapy in human oral squamous cell carcinoma cell lines.

    PubMed

    Moon, Sook; Bae, Jung Yoon; Son, Hwa-Kyung; Lee, Doo Young; Park, Gyeongju; You, Hyun; Ko, Hyojin; Kim, Yong-Chul; Kim, Jin

    2015-02-01

    Photodynamic therapy (PDT) with photosensitizer is one of the promising modalities for cancer treatment. For clinical use of PDT, screening process should be preceded to enhance sensitivity to PDT. Thus, we investigated a molecular biomarker to determine the sensitivity to pheophorbide a (Pa)-PDT in immortalized human oral keratinocytes (IHOK) and oral squamous cell carcinoma (OSCC) cell lines. Two IHOK and several OSCC cell lines were used. After Pa-PDT, cell viability was reduced by more than 50%, and reactive oxygen species were generated in IHOK and OSCC cell lines. Additionally, apoptosis occurred in PDT-treated cells. IHOK(S) and IHOK(P), the two IHOK cell lines derived from the same source, showed a difference in cytotoxicity after Pa-PDT. To explain this difference in cytotoxicity, we looked at the expression of Wnt signaling-related genes in these two cell lines, for the morphology of IHOK(S) which was spindle like and elongated and distinct from IHOK(P) and the parent cell. Among the relevant genes, runt-related transcription factor 3 (RUNX3), an apoptosis-related gene, was selected as a potential marker that confers sensitivity to PDT. We found that the cytotoxicity by Pa-PDT was proportional to RUNX3 expression in OSCC cell lines. Additionally, knockdown of RUNX3 expression reduced cytotoxicity by Pa-PDT, suggesting that RUNX3 might be a biomarker to determine sensitivity to Pa-PDT. This was the first study to find a new target molecule that enhances Pa-PDT effects in IHOK and OSCC cell lines. Hence, the development of a PDT-dependent biomarker could provide a novel approach to improve the effects of PDT on oral precancerous and cancerous lesions.

  3. Suppression of Aedes albopictus, the Asian tiger mosquito, using a 'hot spot' approach.

    PubMed

    Unlu, Isik; Klingler, Kim; Indelicato, Nicholas; Faraji, Ary; Strickman, Daniel

    2016-07-01

    Recent changes in climate and human behavior have led to dramatic increases in the abundance and geographic expansion of invasive mosquito vectors such as Aedes albopictus. Although source reduction has been shown to be effective in reducing mosquito populations, thousands of backyards need to be inspected during door-to-door campaigns, which is labor intensive and expensive. We identified 'hot spots' as high (five or more female or male Ae. albopictus) adult mosquito populations at very focal locations. We tested whether hot spot source reduction efforts were effective in reducing mosquito populations in the early summer season (June to July). Analysis of historical data from the study sites indicated the proportion of hot spots in the control site relative to the intervention site was much greater in 2011, when hot spot treatments were applied to the intervention site, than in 2012, 2013 and 2014 combined, when no sites were treated [OR (95% CI) = 3.9 (1.8, 8.5), Z = 3.39, P < 0.001). Hot spot treatments can be incorporated into existing integrated mosquito management programs to increase effectiveness while reducing the time, cost and effort spent on methods such as door-to-door source reduction. Published 2015. This article is a U.S. Government work and is in the public domain in the USA. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

  4. Single-cell lineage tracking analysis reveals that an established cell line comprises putative cancer stem cells and their heterogeneous progeny

    PubMed Central

    Sato, Sachiko; Rancourt, Ann; Sato, Yukiko; Satoh, Masahiko S.

    2016-01-01

    Mammalian cell culture has been used in many biological studies on the assumption that a cell line comprises putatively homogeneous clonal cells, thereby sharing similar phenotypic features. This fundamental assumption has not yet been fully tested; therefore, we developed a method for the chronological analysis of individual HeLa cells. The analysis was performed by live cell imaging, tracking of every single cell recorded on imaging videos, and determining the fates of individual cells. We found that cell fate varied significantly, indicating that, in contrast to the assumption, the HeLa cell line is composed of highly heterogeneous cells. Furthermore, our results reveal that only a limited number of cells are immortal and renew themselves, giving rise to the remaining cells. These cells have reduced reproductive ability, creating a functionally heterogeneous cell population. Hence, the HeLa cell line is maintained by the limited number of immortal cells, which could be putative cancer stem cells. PMID:27003384

  5. The impact of temperature and Wolbachia infection on vector competence of potential dengue vectors Aedes aegypti and Aedes albopictus in the transmission of dengue virus serotype 1 in southern Taiwan.

    PubMed

    Tsai, Cheng-Hui; Chen, Tien-Huang; Lin, Cheo; Shu, Pei-Yun; Su, Chien-Ling; Teng, Hwa-Jen

    2017-11-07

    We evaluated the impact of temperature and Wolbachia infection on vector competence of the local Aedes aegypti and Ae. albopictus populations of southern Taiwan in the laboratory. After oral infection with dengue serotype 1 virus (DENV-1), female mosquitoes were incubated at temperatures of 10, 16, 22, 28 and 34 °C. Subsequently, salivary gland, head, and thorax-abdomen samples were analyzed for their virus titer at 0, 5, 10, 15, 20, 25 and 30 days post-infection (dpi) by real-time RT-PCR. The results showed that Ae. aegypti survived significantly longer and that dengue viral genome levels in the thorax-abdomen (10 3.25 ± 0.53 -10 4.09 ± 0.71 PFU equivalents/ml) and salivary gland samples (10 2.67 ± 0.33 -10 3.89 ± 0.58 PFU equivalents/ml) were significantly higher at high temperature (28-34 °C). The survival of Ae. albopictus was significantly better at 16 or 28 °C, but the virus titers from thorax-abdomen (10 0.70 -10 2.39 ± 1.31 PFU equivalents/ml) and salivary gland samples (10 0.12 ± 0.05 -10 1.51 ± 0.31 PFU equivalents/ml) were significantly higher at 22-28 °C. Within viable temperature ranges, the viruses were detectable after 10 dpi in salivary glands and head tissues in Ae. aegypti and after 5-10 dpi in Ae. albopictus. Vector competence was measured in Ae. albopictus with and without Wolbachia at 28 °C. Wolbachia-infected mosquitoes survived significantly better and carried lower virus titers than Wolbachia-free mosquitoes. Wolbachia coinfections (92.8-97.2%) with wAlbA and wAlbB strains were commonly found in a wild population of Ae. albopictus. In southern Taiwan, Ae. aegypti is the main vector of dengue and Ae. albopictus has a non-significant role in the transmission of dengue virus due to the high prevalence of Wolbachia infection in the local mosquito population of southern Taiwan.

  6. Uveal Melanoma Cell Lines: Where do they come from? (An American Ophthalmological Society Thesis).

    PubMed

    Jager, Martine J; Magner, J Antonio Bermudez; Ksander, Bruce R; Dubovy, Sander R

    2016-08-01

    To determine whether some of the most often used uveal melanoma cell lines resemble their original tumor. Analysis of the literature, patient charts, histopathology, mutations, chromosome status, HLA type, and expression of melanocyte markers on cell lines and their primary tumors. We examined five cell lines and the primary tumors from which they were derived. Four of the five examined primary tumors were unusual: one occupied the orbit, two were recurrences after prior irradiation, and one developed in an eye with a nevus of Ota. One cell line did not contain the GNA11 mutation, but it was present in the primary tumor. Three of the primary tumors had monosomy 3 (two of these lacked BAP1 expression); however, all five cell lines showed disomy 3 and BAP1 expression. All of the cell lines had gain of 8q. Two cell lines lacked expression of melanocyte markers, although these were present in the corresponding primary tumor. All cell lines could be traced back to their original uveal melanoma. Four of the five primary tumors were unusual. Cell lines often differed from their primary tumor in chromosome status and melanocyte markers. However, their specific chromosome aberrations and capacity to continue proliferation characterize them as uveal melanoma cell lines.

  7. Uveal Melanoma Cell Lines: Where do they come from? (An American Ophthalmological Society Thesis)

    PubMed Central

    Jager, Martine J.; Magner, J. Antonio Bermudez; Ksander, Bruce R.; Dubovy, Sander R.

    2016-01-01

    Purpose To determine whether some of the most often used uveal melanoma cell lines resemble their original tumor. Methods Analysis of the literature, patient charts, histopathology, mutations, chromosome status, HLA type, and expression of melanocyte markers on cell lines and their primary tumors. We examined five cell lines and the primary tumors from which they were derived. Results Four of the five examined primary tumors were unusual: one occupied the orbit, two were recurrences after prior irradiation, and one developed in an eye with a nevus of Ota. One cell line did not contain the GNA11 mutation, but it was present in the primary tumor. Three of the primary tumors had monosomy 3 (two of these lacked BAP1 expression); however, all five cell lines showed disomy 3 and BAP1 expression. All of the cell lines had gain of 8q. Two cell lines lacked expression of melanocyte markers, although these were present in the corresponding primary tumor. Conclusions All cell lines could be traced back to their original uveal melanoma. Four of the five primary tumors were unusual. Cell lines often differed from their primary tumor in chromosome status and melanocyte markers. However, their specific chromosome aberrations and capacity to continue proliferation characterize them as uveal melanoma cell lines. PMID:28018010

  8. Comparative study of the photodynamic effect in tumor and nontumor animal cell lines

    NASA Astrophysics Data System (ADS)

    Stoykova, Elena V.; Alexandrova, R.; Shurulinkov, Stanislav; Sabotinov, O.; Minchev, Georgi

    2004-09-01

    In this study we evaluate the cytotoxicity of two photosensitisers with absorption peaks in the green and red part of the spectrum on animal cell lines. The cytotoxicity assessment was performed for a tumor cell line LSCC-SF-Mc29, obtained from a transplantable chicken hepatoma induced by the myelocytomatosis virus Mc29, a tumor line LSR-SF-SR, obtained from a transplantable sarcoma in rat induced by Rous sarcoma virus strain Schmidt-Ruppin and for normal mouse and bovine cell lines. Up to now the effect of the photodynamic therapy on virus-induced cancers has not been clarified. The cells were treated with 5,10,15,20 - tetra (4-sulfophenyl) porphyrin with main absorption peak at 519 nm and a dye activated with a red light. The cells were seeded in 96-well plates at 2 x 104 cells/well. The cells were exposed to irradiation from a pulsed CuBr vapor laser at 510.6 nm and 578.2 nm and exposure rate 50 mW/cm2, from an Ar-ion laser at 514 nm and 1 mW/cm2 and to 655 nm-irradiation from a semiconductor laser at 10 mW/cm2. The biological activity of the tested compounds was measured by the neutral red uptake cytotoxicity test. The light dose-response curves and light exposures that ensure 50% drop in the treated cells viability in comparison with the cells grown in non-modified medium were obtained for each cell line. The cytotoxic effect of both photosensitisers is most distinguished for the tumor line LSCC-SF-Mc29. The 2-4 times higher viability of the normal cell lines in comparison with the tumor lines is established. The bovine cell lines are more vulnerable than the mouse lines.

  9. Tiger on the prowl: Invasion history and spatio-temporal genetic structure of the Asian tiger mosquito Aedes albopictus (Skuse 1894) in the Indo-Pacific

    PubMed Central

    Ambrose, Luke; Cooper, Robert D.; Chow, Weng K.; Davis, Joseph B.; Muzari, Mutizwa O.; van den Hurk, Andrew F.; Hall-Mendelin, Sonja; Hasty, Jeomhee M.; Burkot, Thomas R.; Bangs, Michael J.; Reimer, Lisa J.; Butafa, Charles; Lobo, Neil F.; Syafruddin, Din; Maung Maung, Yan Naung; Ahmad, Rohani; Beebe, Nigel W.

    2017-01-01

    Background Within the last century, increases in human movement and globalization of trade have facilitated the establishment of several highly invasive mosquito species in new geographic locations with concurrent major environmental, economic and health consequences. The Asian tiger mosquito, Aedes albopictus, is an extremely invasive and aggressive daytime-biting mosquito that is a major public health threat throughout its expanding range. Methodology/Principal findings We used 13 nuclear microsatellite loci (on 911 individuals) and mitochondrial COI sequences to gain a better understanding of the historical and contemporary movements of Ae. albopictus in the Indo-Pacific region and to characterize its population structure. Approximate Bayesian computation (ABC) was employed to test competing historical routes of invasion of Ae. albopictus within the Southeast (SE) Asian/Australasian region. Our ABC results show that Ae. albopictus was most likely introduced to New Guinea via mainland Southeast Asia, before colonizing the Solomon Islands via either Papua New Guinea or SE Asia. The analysis also supported that the recent incursion into northern Australia’s Torres Strait Islands was seeded chiefly from Indonesia. For the first time documented in this invasive species, we provide evidence of a recently colonized population (the Torres Strait Islands) that has undergone rapid temporal changes in its genetic makeup, which could be the result of genetic drift or represent a secondary invasion from an unknown source. Conclusions/Significance There appears to be high spatial genetic structure and high gene flow between some geographically distant populations. The species' genetic structure in the region tends to favour a dispersal pattern driven mostly by human movements. Importantly, this study provides a more widespread sampling distribution of the species’ native range, revealing more spatial population structure than previously shown. Additionally, we present the

  10. Tiger on the prowl: Invasion history and spatio-temporal genetic structure of the Asian tiger mosquito Aedes albopictus (Skuse 1894) in the Indo-Pacific.

    PubMed

    Maynard, Andrew J; Ambrose, Luke; Cooper, Robert D; Chow, Weng K; Davis, Joseph B; Muzari, Mutizwa O; van den Hurk, Andrew F; Hall-Mendelin, Sonja; Hasty, Jeomhee M; Burkot, Thomas R; Bangs, Michael J; Reimer, Lisa J; Butafa, Charles; Lobo, Neil F; Syafruddin, Din; Maung Maung, Yan Naung; Ahmad, Rohani; Beebe, Nigel W

    2017-04-01

    Within the last century, increases in human movement and globalization of trade have facilitated the establishment of several highly invasive mosquito species in new geographic locations with concurrent major environmental, economic and health consequences. The Asian tiger mosquito, Aedes albopictus, is an extremely invasive and aggressive daytime-biting mosquito that is a major public health threat throughout its expanding range. We used 13 nuclear microsatellite loci (on 911 individuals) and mitochondrial COI sequences to gain a better understanding of the historical and contemporary movements of Ae. albopictus in the Indo-Pacific region and to characterize its population structure. Approximate Bayesian computation (ABC) was employed to test competing historical routes of invasion of Ae. albopictus within the Southeast (SE) Asian/Australasian region. Our ABC results show that Ae. albopictus was most likely introduced to New Guinea via mainland Southeast Asia, before colonizing the Solomon Islands via either Papua New Guinea or SE Asia. The analysis also supported that the recent incursion into northern Australia's Torres Strait Islands was seeded chiefly from Indonesia. For the first time documented in this invasive species, we provide evidence of a recently colonized population (the Torres Strait Islands) that has undergone rapid temporal changes in its genetic makeup, which could be the result of genetic drift or represent a secondary invasion from an unknown source. There appears to be high spatial genetic structure and high gene flow between some geographically distant populations. The species' genetic structure in the region tends to favour a dispersal pattern driven mostly by human movements. Importantly, this study provides a more widespread sampling distribution of the species' native range, revealing more spatial population structure than previously shown. Additionally, we present the most probable invasion history of this species in the Australasian

  11. Area-wide management of Aedes albopictus: II. Gauging the efficacy of traditional integrated pest control measures against urban container mosquitoes

    USDA-ARS?s Scientific Manuscript database

    Aedes (Stegomyia) albopictus (Skuse), the Asian tiger mosquito, is an introduced invasive species in the U.S. responsible for a significant proportion of service requests to local mosquito control programs. This container-utilizing mosquito is refractory to standard mosquito abatement measures in th...

  12. GS-nitroxide (JP4-039)-mediated radioprotection of human Fanconi anemia cell lines.

    PubMed

    Bernard, Mark E; Kim, Hyun; Berhane, Hebist; Epperly, Michael W; Franicola, Darcy; Zhang, Xichen; Houghton, Frank; Shields, Donna; Wang, Hong; Bakkenist, Christopher J; Frantz, Marie-Celine; Forbeck, Erin M; Goff, Julie P; Wipf, Peter; Greenberger, Joel S

    2011-11-01

    Fanconi anemia (FA) is an inherited disorder characterized by defective DNA repair and cellular sensitivity to DNA crosslinking agents. Clinically, FA is associated with high risk for marrow failure, leukemia and head and neck squamous cell carcinoma (HNSCC). Radiosensitivity in FA patients compromises the use of total-body irradiation for hematopoietic stem cell transplantation and radiation therapy for HNSCC. A radioprotector for the surrounding tissue would therefore be very valuable during radiotherapy for HNSCC. Clonogenic radiation survival curves were determined for pre- or postirradiation treatment with the parent nitroxide Tempol or JP4-039 in cells of four FA patient-derived cell lines and two transgene-corrected subclonal lines. FancG(-/-) (PD326) and FancD2(-/-) (PD20F) patient lines were more sensitive to the DNA crosslinking agent mitomycin C (MMC) than their transgene-restored subclonal cell lines (both P < 0.0001). FancD2(-/-) cells were more radiosensitive than the transgene restored subclonal cell line (ñ = 2.0 ± 0.7 and 4.7 ± 2.2, respectively, P = 0.03). In contrast, FancG(-/-) cells were radioresistant relative to the transgene-restored subclonal cell line (ñ = 9.4 ± 1.5 and 2.2 ± 05, respectively, P = 0.001). DNA strand breaks measured by the comet assay correlated with radiosensitivity. Cell lines from a Fanc-C and Fanc-A patients showed radiosensitivity similar to that of Fanc-D2(-/-) cells. A fluorophore-tagged JP4-039 (BODIPY-FL) analog targeted the mitochondria of the cell lines. Preirradiation or postirradiation treatment with JP4-039 at a lower concentration than Tempol significantly increased the radioresistance and stabilized the antioxidant stores of all cell lines. Tempol increased the toxicity of MMC in FancD2(-/-) cells. These data provide support for the potential clinical use of JP4-039 for normal tissue radioprotection during chemoradiotherapy in FA patients.

  13. Stem/progenitor cell-like properties of desmoglein 3dim cells in primary and immortalized keratinocyte lines.

    PubMed

    Wan, Hong; Yuan, Ming; Simpson, Cathy; Allen, Kirsty; Gavins, Felicity N E; Ikram, Mohammed S; Basu, Subham; Baksh, Nuzhat; O'Toole, Edel A; Hart, Ian R

    2007-05-01

    We showed previously that primary keratinocytes selected for low desmoglein 3 (Dsg3) expression levels exhibited increased colony-forming efficiency and heightened proliferative potential relative to cells with higher Dsg3 expression levels, characteristics consistent with a more "stem/progenitor cell-like" phenotype. Here, we have confirmed that Dsg3(dim) cells derived from cultured primary human adult keratinocytes have comparability with alpha(6)(bri)/CD71(dim) stem cells in terms of colony-forming efficiency. Moreover, these Dsg3(dim) cells exhibit increased reconstituting ability in in vitro organotypic culture on de-epidermalized dermis (DED); they are small, actively cycling cells, and they express elevated levels of various p63 isoforms. In parallel, using the two immortalized keratinocyte cell lines HaCaT and NTERT, we obtained essentially similar though occasionally different findings. Thus, reduced colony-forming efficiency by Dsg3(bri) cells consistently was observed in both cell lines even though the cell cycle profile and levels of p63 isoforms in the bri and dim populations differed between these two cell lines. Dsg3(dim) cells from both immortalized lines produced thicker and better ordered hierarchical structural organization of reconstituted epidermis relative to Dsg3(bri) and sorted control cells. Dsg3(dim) HaCaT cells also show sebocyte-like differentiation in the basal compartment of skin reconstituted after a 4-week organotypic culture. No differences in percentages of side population cells (also a putative marker of stem cells) were detected between Dsg3(dim) and Dsg3(bri) populations. Taken together our data indicate that Dsg3(dim) populations from primary human adult keratinocytes and long-term established keratinocyte lines possess certain stem/progenitor cell-like properties, although the side population characteristic is not one of these features. Disclosure of potential conflicts of interest is found at the end of this article.

  14. Coffee and its waste repel gravid Aedes albopictus females and inhibit the development of their embryos.

    PubMed

    Satho, Tomomitsu; Dieng, Hamady; Ahmad, Muhammad Hishamuddin Itam; Ellias, Salbiah Binti; Hassan, Ahmad Abu; Abang, Fatimah; Ghani, Idris Abd; Miake, Fumio; Ahmad, Hamdan; Fukumitsu, Yuki; Zuharah, Wan Fatma; Majid, Abdul Hafiz Ab; Kassim, Nur Faeza Abu; Hashim, Nur Aida; Ajibola, Olaide Olawunmi; Al-Khayyat, Fatima Abdulla; Nolasco-Hipolito, Cirilo

    2015-05-14

    Dengue is a prevalent arboviral disease and the development of insecticide resistance among its vectors impedes endeavors to control it. Coffee is drunk by millions of people daily worldwide, which is associated with the discarding of large amounts of waste. Coffee and its waste contain large amounts of chemicals many of which are highly toxic and none of which have a history of resistance in mosquitoes. Once in solution, coffee is brownish in colour, resembling leaf infusion, which is highly attractive to gravid mosquitoes. To anticipate the environmental issues related to the increasing popularity of coffee as a drink, and also to combat insecticide resistance, we explored the deterrence potentials of coffee leachates against the ovipositing and embryonic stages of the dengue vector, Aedes albopictus. In a series of choice, no-choice, and embryo toxicity bioassays, we examined changes in the ovipositional behaviours and larval eclosion of Ae. albopictus in response to coffee extracts at different concentrations. Oviposition responses were extremely low when ovicups holding highly concentrated extract (HCE) of coffee were the only oviposition sites. Gravid females retained increased numbers of mature eggs until 5 days post-blood feeding. When provided an opportunity to oviposit in cups containing coffee extracts and with water, egg deposition occurred at lower rates in those containing coffee, and HCE cups were far less attractive to females than those containing water only. Females that successfully developed in a coffee environment preferentially oviposited in such cups when in competition with preferred oviposition sites (water cups), but this trait did not continue into the fourth generation. Larval eclosion occurred at lower rates among eggs that matured in a coffee environment, especially among those that were maintained on HCE-moistened substrates. The observations of the present study indicate a pronounced vulnerability of Ae. albopictus to the presence

  15. Reintroduction of the invasive mosquito species Aedes albopictus in Belgium in July 2013.

    PubMed

    Boukraa, Slimane; Raharimalala, Fara N; Zimmer, Jean-Yves; Schaffner, Francis; Bawin, Thomas; Haubruge, Eric; Francis, Frédéric

    2013-01-01

    Since its first report in 2000, the invasive mosquito Aedes albopictus was not found any more during the different entomological inspections performed at its place of introduction in Belgium between 2001 and 2012. In July 2013, one adult male was captured at the same site (a platform of imported used tires located in Vrasene, Oost-Vlaanderen Province), during a monitoring using CO2-baited trap. This finding suggests the reintroduction of the species in Belgium via the used tire trade. © S. Boukraa et al., published by EDP Sciences, 2013.

  16. Control of Aedes albopictus with attractive toxic sugar baits (ATSB) and potential impact on non-target organisms in St. Augustine, Florida

    PubMed Central

    Revay, Edita E.; Müller, Gunter C.; Qualls, Whitney A.; Kline, Daniel; Naranjo, Diana P.; Arheart, Kristopher L.; Kravchenko, Vasiliy D.; Yfremova, Zoya; Hausmann, Axel; Beier, John C.; Schlein, Yosef; Xue, Rui-De

    2013-01-01

    The purpose of this study was to test the efficacy of bait stations and foliar applications containing attractive toxic sugar baits (ATSB) and eugenol to control Aedes albopictus. At the same time the potential impact of these control methods was evaluated on non-target organisms. The study was conducted at five tire sites in St. Augustine, Florida. Aedes albopictus populations were significantly reduced with ATSB-eugenol applications applied directly to non-flowering vegetation and as bait stations compared with non-attractive sugar baits and control. The application of ATSB made to non-flowering vegetation resulted in more significant reductions of mosquito populations compared to the application of ATSB presented in a bait station. Over 5.5% of the non-targets were stained in the flowering vegetation application site. However, when the attractive sugar bait application was made to non-flowering vegetation or presented in bait stations the impact on non-target insects was very low for all non-target orders as only 0.6% of the individual insects were stained with the dye from the sugar solutions, respectively. There were no significant differences between the staining of mosquitoes collected in flowering vegetation (206/1000) or non-flowering vegetation (242/1000) sites during the non-target evaluation. Our field studies support the use of eugenol as an active ingredient for controlling the dengue vector Ae. albopictus when used as an ATSB toxin and demonstrates potential use in sub-tropical and tropical environments for dengue control. PMID:24122115

  17. Larval Mosquito Habitat Utilization and Community Dynamics of Aedes albopictus and Aedes japonicus (Diptera: Culicidae)

    DTIC Science & Technology

    2012-01-01

    Medical Entomology, 49(4):813-824. 2012. Published By: Entomological Society of America DOI: http://dx.doi.org/10.1603/ME11031 URL: http...importance as a disease vector of dengue and dengue hemorrhagic fever (Estrada-Franco and Craig 1995, Knudsen et al. 1996).Ae. albopictuswas solely...respon- sible from the outbreak of dengue fever in Hawaii during 2001 (Efßer et al. 2005). Ae. albopictus is also a potential vector of Venezuelan equine

  18. The neuroendocrine phenotype, genomic profile and therapeutic sensitivity of GEPNET cell lines

    PubMed Central

    Hofving, Tobias; Arvidsson, Yvonne; Almobarak, Bilal; Inge, Linda; Pfragner, Roswitha; Persson, Marta; Stenman, Göran; Kristiansson, Erik; Johanson, Viktor; Nilsson, Ola

    2018-01-01

    Experimental models of neuroendocrine tumour disease are scarce, and no comprehensive characterisation of existing gastroenteropancreatic neuroendocrine tumour (GEPNET) cell lines has been reported. In this study, we aimed to define the molecular characteristics and therapeutic sensitivity of these cell lines. We therefore performed immunophenotyping, copy number profiling, whole-exome sequencing and a large-scale inhibitor screening of seven GEPNET cell lines. Four cell lines, GOT1, P-STS, BON-1 and QGP-1, displayed a neuroendocrine phenotype while three others, KRJ-I, L-STS and H-STS, did not. Instead, these three cell lines were identified as lymphoblastoid. Characterisation of remaining authentic GEPNET cell lines by copy number profiling showed that GOT1, among other chromosomal alterations, harboured losses on chromosome 18 encompassing the SMAD4 gene, while P-STS had a loss on 11q. BON-1 had a homozygous loss of CDKN2A and CDKN2B, and QGP-1 harboured amplifications of MDM2 and HMGA2. Whole-exome sequencing revealed both disease-characteristic mutations (e.g. ATRX mutation in QGP-1) and, for patient tumours, rare genetic events (e.g. TP53 mutation in P-STS, BON-1 and QGP-1). A large-scale inhibitor screening showed that cell lines from pancreatic NETs to a greater extent, when compared to small intestinal NETs, were sensitive to inhibitors of MEK. Similarly, neuroendocrine NET cells originating from the small intestine were considerably more sensitive to a group of HDAC inhibitors. Taken together, our results provide a comprehensive characterisation of GEPNET cell lines, demonstrate their relevance as neuroendocrine tumour models and explore their therapeutic sensitivity to a broad range of inhibitors. PMID:29444910

  19. CD40 expression in Wehi-164 cell line

    PubMed Central

    Ebadi, Padideh; Pourfathollah, Ali Akbar; Soheili, Zahra Soheila; Moazzeni, Seyed Mohammad

    2010-01-01

    CD40-CD154 interaction is an important process for cellular and humoral immunity regulation and can be effective in the body’s defense against tumors. In the present study, we evaluated the expression of CD40 in Wehi-164 cell line. CD40 expressions on the cell surface and in the cytoplasm were assessed by flow cytometry and intracellular staining assay, respectively. Also, the mRNA expression was identified by real time-PCR. The obtained results showed the high mRNA and cytoplasmic protein expression of CD40 but no surface expression. These results suggest that the Wehi-164 cell line down regulates expression of CD40 on the surface for evasion of immune system. PMID:20496113

  20. CD40 expression in Wehi-164 cell line.

    PubMed

    Karimi, Mohammad Hossein; Ebadi, Padideh; Pourfathollah, Ali Akbar; Soheili, Zahra Soheila; Moazzeni, Seyed Mohammad

    2010-07-01

    CD40-CD154 interaction is an important process for cellular and humoral immunity regulation and can be effective in the body's defense against tumors. In the present study, we evaluated the expression of CD40 in Wehi-164 cell line. CD40 expressions on the cell surface and in the cytoplasm were assessed by flow cytometry and intracellular staining assay, respectively. Also, the mRNA expression was identified by real time-PCR. The obtained results showed the high mRNA and cytoplasmic protein expression of CD40 but no surface expression. These results suggest that the Wehi-164 cell line down regulates expression of CD40 on the surface for evasion of immune system.

  1. Heterogeneity of osteosarcoma cell lines led to variable responses in reprogramming.

    PubMed

    Choong, Pei Feng; Teh, Hui Xin; Teoh, Hoon Koon; Ong, Han Kiat; Choo, Kong Bung; Sugii, Shigeki; Cheong, Soon Keng; Kamarul, Tunku

    2014-01-01

    Four osteosarcoma cell lines, Saos-2, MG-63, G-292 and U-2 OS, were reprogrammed to pluripotent state using Yamanaka factors retroviral transduction method. Embryonic stem cell (ESC)-like clusters started to appear between 15 to 20 days post transduction. Morphology of the colonies resembled that of ESC colonies with defined border and tightly-packed cells. The reprogrammed sarcomas expressed alkaline phosphatase and pluripotency markers, OCT4, SSEA4, TRA-1-60 and TRA-1-81, as in ESC up to Passage 15. All reprogrammed sarcomas could form embryoid body-like spheres when cultured in suspension in a low attachment dish for up to 10 days. Further testing on the directed differentiation capacity of the reprogrammed sarcomas showed all four reprogrammed sarcoma lines could differentiate into adipocytes while reprogrammed Saos-2-REP, MG-63-REP and G-292-REP could differentiate into osteocytes. Among the 4 osteosarcoma cell lines, U-2 OS reported the highest transduction efficiency but recorded the lowest reprogramming stability under long term culture. Thus, there may be intrinsic differences governing the variable responses of osteosarcoma cell lines towards reprogramming and long term culture effect of the reprogrammed cells. This is a first report to associate intrinsic factors in different osteosarcoma cell lines with variable reprogramming responses and effects on the reprogrammed cells after prolonged culture.

  2. Infectivity of Sf-rhabdovirus variants in insect and mammalian cell lines.

    PubMed

    Maghodia, Ajay B; Jarvis, Donald L

    2017-12-01

    Sf-rhabdovirus was only recently identified as an adventitious agent of Spodoptera frugiperda (Sf) cell lines used as hosts for baculovirus vectors. As such, we still know little about its genetic variation, infectivity, and the potential impact of variation on the Sf-rhabdovirus-host interaction. Here, we characterized Sf-rhabdoviruses from two widely used Sf cell lines to confirm and extend information on Sf-rhabdovirus variation. We then used our novel Sf-rhabdovirus-negative (Sf-RVN) Sf cell line to assess the infectivity of variants with and without a 320bp X/L deletion and found both established productive persistent infections in Sf-RVN cells. We also assessed their infectivity using heterologous insect and mammalian cell lines and found neither established productive persistent infections in these cells. These results are the first to directly demonstrate Sf-rhabdoviruses are infectious for Sf cells, irrespective of the X/L deletion. They also confirm and extend previous results indicating Sf-rhabdoviruses have a narrow host range. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Production of thrombopoietin (TPO) by rat hepatocytes and hepatoma cell lines.

    PubMed

    Shimada, Y; Kato, T; Ogami, K; Horie, K; Kokubo, A; Kudo, Y; Maeda, E; Sohma, Y; Akahori, H; Kawamura, K

    1995-12-01

    Recently, we purified rat thrombopoietin (TPO) from plasma of irradiated rats (XRP) by measuring its activity that stimulated the production of megakaryocytes from megakaryocyte progenitor cells (CFU-MK) in vitro. We then cloned the cDNAs for rat and human TPO. In this study, we found the production of TPO by hepatocytes isolated with the collagenase perfusion method from both normal and thrombocytopenic rats, by a two-step fractionation of hepatocyte culture medium (CM). Subsequently, CM of rat hepatoma cell lines was screened for the presence of TPO; three cell lines, H4-II-E, McA-RH8994, and HTC, were found to produce TPO. According to the purification procedure for TPO from XRP, TPO was partially purified from 2 L CM of each of three cell lines with a six-step procedure. In the final reverse-phase column, TPO from each cell line was eluted with the same retention time as that from XRP, and the TPO fraction exhibited megakaryocyte colony-stimulating activity (Meg-CSA). TPO-active fraction eluted from the final reverse-phase column was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), extracted from the gel, and assayed. TPO activity from each cell line was found in the respective molecular weight region, indicating the heterogeneity of the TPO molecule. Using reverse transcriptase-polymerase chain reaction (RT-PCR), we detected the expression of TPO mRNA in hepatocytes, three hepatoma cell lines, normal rat liver, and X-irradiated rat liver. Northern blot analysis showed that TPO mRNA was expressed mainly in liver among the various organs tested. These data demonstrate that TPO is produced by rat hepatocytes and hepatoma cell lines and suggest that liver may be the primary organ that produces TPO.

  4. Analysis of differential protein expression in normal and neoplastic human breast epithelial cell lines

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Williams, K.; Chubb, C.; Huberman, E.

    High resolution two dimensional get electrophoresis (2DE) and database analysis was used to establish protein expression patterns for cultured normal human mammary epithelial cells and thirteen breast cancer cell lines. The Human Breast Epithelial Cell database contains the 2DE protein patterns, including relative protein abundances, for each cell line, plus a composite pattern that contains all the common and specifically expressed proteins from all the cell lines. Significant differences in protein expression, both qualitative and quantitative, were observed not only between normal cells and tumor cells, but also among the tumor cell lines. Eight percent of the consistently detected proteinsmore » were found in significantly (P < 0.001) variable levels among the cell lines. Using a combination of immunostaining, comigration with purified protein, subcellular fractionation, and amino-terminal protein sequencing, we identified a subset of the differentially expressed proteins. These identified proteins include the cytoskeletal proteins actin, tubulin, vimentin, and cytokeratins. The cell lines can be classified into four distinct groups based on their intermediate filament protein profile. We also identified heat shock proteins; hsp27, hsp60, and hsp70 varied in abundance and in some cases in the relative phosphorylation levels among the cell lines. Finally, we identified IMP dehydrogenase in each of the cell lines, and found the levels of this enzyme in the tumor cell lines elevated 2- to 20-fold relative to the levels in normal cells.« less

  5. Immortalized human hepatic cell lines for in vitro testing and research purposes

    PubMed Central

    Ramboer, Eva; Vanhaecke, Tamara; Rogiers, Vera; Vinken, Mathieu

    2015-01-01

    Summary The ubiquitous shortage of primary human hepatocytes has urged the scientific community to search for alternative cell sources, such as immortalized hepatic cell lines. Over the years, several human hepatic cell lines have been produced, whether or not using a combination of viral oncogenes and human telomerase reverse transcriptase protein. Conditional approaches for hepatocyte immortalization have also been established and allow generation of growth-controlled cell lines. A variety of immortalized human hepatocytes have already proven useful as tools for liver-based in vitro testing and fundamental research purposes. The present chapter describes currently applied immortalization strategies and provides an overview of the actually available immortalized human hepatic cell lines and their in vitro applications. PMID:26272134

  6. Immortalized Human Hepatic Cell Lines for In Vitro Testing and Research Purposes.

    PubMed

    Ramboer, Eva; Vanhaecke, Tamara; Rogiers, Vera; Vinken, Mathieu

    2015-01-01

    The ubiquitous shortage of primary human hepatocytes has urged the scientific community to search for alternative cell sources, such as immortalized hepatic cell lines. Over the years, several human hepatic cell lines have been produced, whether or not using a combination of viral oncogenes and human telomerase reverse transcriptase protein. Conditional approaches for hepatocyte immortalization have also been established and allow generation of growth-controlled cell lines. A variety of immortalized human hepatocytes have already proven useful as tools for liver-based in vitro testing and fundamental research purposes. The present chapter describes currently applied immortalization strategies and provides an overview of the actually available immortalized human hepatic cell lines and their in vitro applications.

  7. Heterogeneity in cancer cells: variation in drug response in different primary and secondary colorectal cancer cell lines in vitro.

    PubMed

    Arul, Melanie; Roslani, April Camilla; Cheah, Swee Hung

    2017-05-01

    Tumor heterogeneity may give rise to differential responses to chemotherapy drugs. Therefore, unraveling tumor heterogeneity has an implication for biomarker discovery and cancer therapeutics. To test this phenomenon, we investigated the differential responses of three secondary colorectal cancer cell lines of different origins (HCT116, HT29, and SW620 cells) and four novel primary cell lines obtained from different colorectal cancer patients to 5-fluorouracil (5-FU) and oxaliplatin (L-OHP) and explored the differences in gene expression among the primary cell lines in response to exposure to cytotoxic drugs. Cells were exposed to different doses of 5-FU and L-OHP separately or in combinations of equitoxic drug or equimolar drug ratios (median effect of Chou-Talalay principle). Cell viability was assessed using MTT assay and the respective IC 50 values were determined. Changes in gene expression in primary cell lines after exposure to the same drug doses were compared using real-time PCR array. The sensitivities (IC 50 ) of different cell lines, both secondary and primary, to 5-FU and L-OHP were significantly different, whether in monotherapy or combined treatment. Primary cell lines needed higher doses to reach IC 50 . There were variations in gene expression among the primary cell lines of different chemosensitivities to the challenge of the same combined dose of 5-FU and L-OHP. The results confirm the heterogeneous nature of colorectal cancer cells from different patient tumors. Studies using primary cancer cells established from patient's tumors rather than secondary cell lines will more closely reflect the actual character of the disease.

  8. 9 CFR 113.52 - Requirements for cell lines used for production of biologics.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Requirements for cell lines used for... STANDARD REQUIREMENTS Ingredient Requirements § 113.52 Requirements for cell lines used for production of... cell line used to prepare a biological product shall be tested as prescribed in this section. A cell...

  9. 9 CFR 113.52 - Requirements for cell lines used for production of biologics.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Requirements for cell lines used for... STANDARD REQUIREMENTS Ingredient Requirements § 113.52 Requirements for cell lines used for production of... cell line used to prepare a biological product shall be tested as prescribed in this section. A cell...

  10. 9 CFR 113.52 - Requirements for cell lines used for production of biologics.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Requirements for cell lines used for... STANDARD REQUIREMENTS Ingredient Requirements § 113.52 Requirements for cell lines used for production of... cell line used to prepare a biological product shall be tested as prescribed in this section. A cell...

  11. 9 CFR 113.52 - Requirements for cell lines used for production of biologics.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Requirements for cell lines used for... STANDARD REQUIREMENTS Ingredient Requirements § 113.52 Requirements for cell lines used for production of... cell line used to prepare a biological product shall be tested as prescribed in this section. A cell...

  12. 9 CFR 113.52 - Requirements for cell lines used for production of biologics.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Requirements for cell lines used for... STANDARD REQUIREMENTS Ingredient Requirements § 113.52 Requirements for cell lines used for production of... cell line used to prepare a biological product shall be tested as prescribed in this section. A cell...

  13. Targeting a Hidden Enemy: Pyriproxyfen Autodissemination Strategy for the Control of the Container Mosquito Aedes albopictus in Cryptic Habitats

    PubMed Central

    Suman, Devi Shankar; Wang, Yi; Unlu, Isik; Williges, Eric; Williams, Gregory M.; Gaugler, Randy

    2016-01-01

    Background The Asian tiger mosquito, Aedes albopictus, is a vector of dengue, Chikungunya, and Zika viruses. This mosquito inhabits a wide range of artificial water-holding containers in urban and suburban areas making it difficult to control. We tested the hypothesis that female-driven autodissemination of an insect growth regulator could penetrate cryptic oviposition habitats difficult to treat with conventional insecticidal sprays. Methodology Oviposition preferences of Ae. albopictus females for open and cryptic cups were tested in semi-field experiments. Two conventional larvicidal sprayers were tested to determine droplet penetration and larvicidal efficacy in open and cryptic habitats using Bacillus thuringiensis var. israelensis (Bti) in the field. Finally, the efficacy of pyriproxyfen autodissemination stations was assessed in cryptic and open cups in residential areas during 2013 and 2014. Principal Findings Gravid females strongly preferred cryptic (53.1±12.9 eggs/cup) over open (10.3±4.3 eggs/cup) cups for oviposition. Cryptic cups showed limited droplet penetration and produced 0.1–0.3% larval mortality with a conventional backpack and low-volume sprays of Bti. The autodissemination stations effectively contaminated these cryptic cups (59.3–84.6%) and produced 29.7–40.8% pupal mortality during 2013–2014. Significant pupal mortality was also observed in open cups. Conclusions The autodissemination station effectively exploits the oviposition behavior of wild gravid females to deliver pyriproxyfen to targeted oviposition habitats. Although the pupal mortality in cryptic cups was relatively lower than expected for the effective vector control. Autodissemination approach may be a suitable supporting tool to manage Ae. albopictus immatures in the cryptic habitats those are less accessible to conventional larvicidal sprays. PMID:28033379

  14. MiRNA-Target Interaction Reveals Cell-Specific Post-Transcriptional Regulation in Mammalian Cell Lines

    PubMed Central

    Kulkarni, Varun; Naqvi, Afsar Raza; Uttamani, Juhi Raju; Nares, Salvador

    2016-01-01

    MicroRNAs are 18–22 nucleotides long, non-coding RNAs that bind transcripts with complementary sequences leading to either mRNA degradation or translational suppression. However, the inherent differences in preferred mode of miRNA regulation among cells of different origin have not been examined. In our previous transcriptome profiling studies, we observed that post-transcriptional regulation can differ substantially depending on the cell in context. Here we examined mechanistic differences in the regulation of a let-7a targeted (wild type) or resistant (mutant) engineered renilla transcript across various mammalian cell lines of diverse origin. Dual luciferase assays show that compared to mutant (mut), the reporter gene containing wild type (wt) let-7a binding sites was efficiently suppressed upon transfection in various cell lines. Importantly, the strength of miRNA regulation varied across the cell lines. Total RNA analysis demonstrates that wt renilla mRNA was expressed to similar or higher levels compared to mut suggesting that translation repression is a predominant mode of miRNA regulation. Nonetheless, transcript degradation was observed in some cell lines. Ago-2 immunoprecipitation show that miRNA repressed renilla mRNA are associated with functional mi-RISC (miRNA-RNA induced silencing complex). Given the immense potential of miRNA as a therapeutic option, these findings highlight the necessity to thoroughly examine the mode of mRNA regulation in order to achieve the beneficial effects in targeting cells. PMID:26761000

  15. Characterization of the Murine Myeloid Precursor Cell Line MuMac-E8

    PubMed Central

    Fricke, Stephan; Riemschneider, Sina; Kohlschmidt, Janine; Hilger, Nadja; Fueldner, Christiane; Knauer, Jens; Sack, Ulrich; Emmrich, Frank; Lehmann, Jörg

    2014-01-01

    Starting point for the present work was the assumption that the cell line MuMac-E8 represents a murine cell population with stem cell properties. Preliminary studies already pointed to the expression of stem-cell associated markers and a self-regenerative potential of the cells. The cell line MuMac-E8 should be examined for their differential stage within stem cell hierarchy. MuMac-E8 cells were derived from a chimeric mouse model of arthritis. It could be shown that MuMac-E8 cells express mRNA of some genes associated with pluripotent stem cells (Nanog, Nucleostemin), of genes for hematopoietic markers (EPCR, Sca-1, CD11b, CD45), for the mesenchymal marker CD105 and of genes for the neural markers Pax-6 and Ezrin. In methylcellulose and May-Grünwald-Giemsa staining, hematopoietic colonies were obtained but the hematopoietic system of lethally irradiated mice could not be rescued. Osteogenic differentiation was not detectable. Thus, it became evident that MuMac-E8 represents not a stem cell line. However, MuMac-E8 cells expressed several myeloid surface markers (i.e. CD11b, F4/80, CD14, CD64), showed phagocytosis and is capable of producing nitric oxide. Thus, this cell line seems to be arrested an advanced stage of myeloid differentiation. Adherence data measured by impedance-based real-time cell analysis together with cell morphology data suggested that MuMac-E8 represents a new macrophage precursor cell line exhibiting weak adherence. This cell line is suitable as an in-vitro model for testing of macrophage functions. Moreover, it might be also useful for differentiation or reprogramming studies. PMID:25546418

  16. Antiproliferative effect of isopentenylated coumarins on several cancer cell lines.

    PubMed

    Kawaii, S; Tomono, Y; Ogawa, K; Sugiura, M; Yano, M; Yoshizawa, Y; Ito, C; Furukawa, H

    2001-01-01

    33 coumarins, mainly the simple isopentenylated coumarins and derived pyrano- and furanocoumarins, were examined for their antiproliferative activity towards several cancer and normal human cell lines. The pyrano- and furanocoumarins showed strong activity against the cancer cell lines, whereas they had weak antiproliferative activity against the normal human cell lines. The decreasing rank order of potency was osthenone (10), clausarin (25), clausenidin (26), dentatin (24), nordentatin (23), imperatorin (29), seselin (27), xanthyletin (21), suberosin (17), phebalosin (8) and osthol (12). The structure-activity relationship established from the results revealed that the 1,1-dimethylallyl and isopentenyl groups have an important role for antiproliferative activity.

  17. SU-C-204-04: Irradiation of Human Cell Lines Using Various Ions

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Lin, Y; McMahon, S; Kaminuma, T

    2016-06-15

    Purpose: The purpose of this study is to investigate and quantify the biological effects of ion radiation using several human cell lines. We aim to answer the question of whether carbon ion the most ideal ion species for heavy ion radiotherapy. Methods: The cells were irradiated at different positions along the pristine Bragg peak of several ions with different atomic number. The biological effectiveness was evaluated using the clonogenic cell survival assay. Irradiation of three human lung cancer cell lines and a fibroblast cell line were undertaken using the charged particle beam at the NASA Space Radiation Laboratory at Brookhavenmore » National Lab. Four mono-energetic ion beams (carbon, oxygen, helium and lithium) were used to irradiate the cells. Water or media-filled T25 flasks were lined up along the beam line so that the cell-containing surfaces of the flasks were placed at a specific depth along the pristine Bragg curve. Four depths along the curve, representing entrance point, rising peak, peak and distal fall off, were selected to determine biological effectiveness. Gaf-chromic films were placed between the flasks to monitor the irradiation as soon as it was finished. Results: For all ion radiations, the maximum cell killing effect occurs at either peak or distal fall off, depending on the cell lines. For instance, for the fibroblast cell line AGO1522, RBEs of 1.4, 1.2, 1.4 and 1.9 were observed at the Bragg peak for Helium, Lithium, Carbon and Oxygen ions. Comparing positions, RBEs of 0.9, 1.2, 1.4 and 1.8 were observed for carbon irradiation of AGO-1522 cells positions corresponding to entrance, rising peak, peak and distal fall off. Conclusion: RBE values differ with position in the Bragg peak, ion species and cell line. Ions other than carbon may prove more effective in certain irradiation conditions and may contribute to optimized heavy ion therapy.« less

  18. Enhanced production of enveloped viruses in BST-2-deficient cell lines.

    PubMed

    Yi, Eunbi; Oh, Jinsoo; Giao, Ngoc Q; Oh, Soohwan; Park, Se-Ho

    2017-10-01

    Despite all the advantages that cell-cultured influenza vaccines have over egg-based influenza vaccines, the inferior productivity of cell-culture systems is a major drawback that must be addressed. BST-2 (tetherin) is a host restriction factor which inhibits budding-out of various enveloped viruses from infected host cells. We developed BST-2-deficient MDCK and Vero cell lines to increase influenza virus release in cell culture. BST-2 gene knock-out resulted in increased release of viral particles into the culture medium, by at least 2-fold and up to 50-fold compared to release from wild-type counterpart cells depending on cell line and virus type. The effect was not influenza virus/MDCK/Vero-specific, but was also present in a broad range of host cells and virus families; we observed similar results in murine, human, canine, and monkey cell lines with viruses including MHV-68 (Herpesviridae), influenza A virus (Orthomyxoviridae), porcine epidemic diarrhea virus (Coronaviridae), and vaccinia virus (Poxviridae). Our results suggest that the elimination of BST-2 expression in virus-producing cell lines can enhance the production of viral vaccines. Biotechnol. Bioeng.2017;114: 2289-2297. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  19. Behavioral responses of two dengue virus vectors, Aedes aegypti and Aedes albopictus (Diptera: Culicidae), to DUET TM and its components

    USDA-ARS?s Scientific Manuscript database

    Ultralow volume (ULV) droplets of DUET TM, prallethrin and sumithrin at a sublethal dose were applied to unfed (non bloodfed) and bloodfed female Aedes aegypti Linn. and Aedes albopictus (Skuse) in a wind tunnel. Control spray droplets only contained inactive ingredients. Individual mosquitoes wer...

  20. Efficacy of Ovitrap Colors and Patterns for Attracting Aedes albopictus (Diptera: Culicidae) at Suburban Field Sites in North Central Florida

    USDA-ARS?s Scientific Manuscript database

    We sought to visually enhance the attractiveness of a standard black ovitrap routinely used in surveillance of the Asian Tiger Mosquito, Aedes albopictus (Skuse), and now being used as lethal ovitraps in Ae. aegypti (L.) dengue control programs. Black plastic drinking cups (ovitraps) were visually ...