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PubMedDate not supplied

Approaches to vector control: new and trusted. 1. Humoral immune responses in blackfly and mosquito vectors of filariae.

The vectors of filariasis, mosquitoes and blackflies, are capable of mounting a defence response to the infection. This selective review describes the molecules that are involved in these immune systems. Several antibacterial peptides are known to be induced and secreted into the haemolymph by the fat body and the circulating haemocytes. In addition, haemagglutinating lectins with carbohydrate specificities to the surface of the developing filarial larvae appear. Activation of a range of proteases occurs rapidly as does activation of the prophenoloxidase pathway. The possible roles of these and other molecules is discussed, together with mention of a working hypothesis as to how these molecules may be regulated.

Animals↗
PubMed1984

Positive selection vectors: a small plasmid vector useful for the direct selection of Sau3A-generated overlapping DNA fragments.

A positive selection plasmid vector ( pLA7 ), containing a unique BclI site, and its use in the facile selection of a library of overlapping DNA fragments, generated by a partial digest with Sau3A, is described. Selection depends on the 5-fluorouracil + 5' AMP resistance of upp- ush - [ pLA7ush ::cloned DNA] cells, whereas upp- ush - [ pLA7ush +] cells are sensitive under the same conditions.

Chromosome Mapping↗
PubMed1987

Vectors with restriction site banks. V. pJRD215, a wide-host-range cosmid vector with multiple cloning sites.

The construction of a new wide-host-range, restriction-site bank, cosmid-cloning vehicle (pJRD215) is described. The wide-host-range properties and the ability to be transferred by conjugation, extend genetic engineering to those Gram-negative species that cannot be transformed. The vector permits the cloning of genes from Gram-negative bacteria using a complementation screening procedure in a mutant host. This procedure is simplified by the possibility of construction of a cosmid gene bank so that only a few hundred clones need to be screened. Subsequent subcloning of the gene of interest is facilitated by the presence of at least 23 unique cloning sites.

Base Sequence↗
PubMed2005

A potyvirus-based gene vector allows producing active human S-COMT and animal GFP, but not human sorcin, in vector-infected plants.

Potato virus A (PVA), a potyvirus with a (+)ssRNA genome translated to a large polyprotein, was engineered and used as a gene vector for expression of heterologous proteins in plants. Foreign genes including jellyfish GFP (Aequorea victoria) encoding the green fluorescent protein (GFP, 27 kDa) and the genes of human origin (Homo sapiens) encoding a soluble resistance-related calcium-binding protein (sorcin, 22 kDa) and the catechol-O-methyltransferase (S-COMT; 25 kDa) were cloned between the cistrons for the viral replicase and coat protein (CP). The inserts caused no adverse effects on viral infectivity and virulence, and the inserted sequences remained intact in progeny viruses in the systemically infected leaves. The heterologous proteins were released from the viral polyprotein following cleavage by the main viral proteinase, NIa, at engineered proteolytic processing sites flanking the insert. Active GFP, as indicated by green fluorescence, and S-COMT with high levels of enzymatic activity were produced. In contrast, no sorcin was detected despite the expected equimolar amounts of the foreign and viral proteins being expressed as a polyprotein. These data reveal inherent differences between heterologous proteins in their suitability for production in plants.

Amino Acid Sequence↗
PubMed2005

Using QPCR to assign infectious potencies to adenovirus based vaccines and vectors for gene therapy: toward a universal method for the facile quantitation of virus and vector potency.

The assignment of infectious potency to test articles of adenovirus has been conducted mainly using classical end-point dilution methods, which rely on virus induced cytopathology to reveal the presence of infectious virus. These assays suffer the disadvantages of labor intensity, duration, throughput restriction and variability. In the course of our development of an Ad5 based HIV vaccine for clinical evaluation, we sought a facile method for the assignment of potency to the numerous test articles generated during the development of bioprocesses for bulk manufacture, downstream purification and formulation. In this paper we describe a quantitative PCR based potency assay (QPA) which uses QPCR to quantitate adenovirus genomes replicated 24h after the inoculation of a test article on 293 cell monolayers, and then relates that mass to potency by interpolation to a standard curve of replicated adenovirus genomes constructed with a reference adenovirus standard to which infectious potency has been previously assigned in the classical end-point dilution assay. The QPA assay for adenovirus is simple and rapid, with a throughput capacity adequate to the potency assay demands of bioprocess development, and with a precision expressed as a root variability of 16.8% R.S.D., allowing for close discriminations of the products of alternative process configurations. The adenovirus QPA principle can be applied to the quantitation of infectious potency of both RNA and DNA viruses and we report briefly on the development of QPA assays for measles and mumps. QPA assays owing to their simplicity and easy automation, rapidity, capacity and precision hold promise to become widely practiced methods for the quantitation of the potency of live virus vaccines and other recombinant virus vectors.

Adenoviridae↗
PubMed1995

Cellular and humoral immune responses to adenoviral vectors containing factor IX gene: tolerization of factor IX and vector antigens allows for long-term expression.

Recombinant adenoviruses containing the canine factor IX (FIX) cDNA were directly introduced in the hind leg muscle of mice. We show that (i) in nude mice, high expression (1-5 micrograms/ml in plasma) of FIX protein can be detected for > 300 days; (ii) in contrast, expression of FIX protein was transient (7-10 days) in normal mice; (iii) CD8+ lymphocytes could be detected within 3 days in the infected muscle tissue; (iv) use of beta 2-microglobulin and immunoglobulin M heavy chain "knockout" mice showed that lack of sustained expression of FIX protein is due to cell-mediated and humoral immune responses; (v) normal mice, once infected with recombinant adenovirus, could not be reinfected efficiently for at least 30 days due to neutralizing viral antibodies; and, finally, (vi) using immunosuppressive drugs, some normal mice can be tolerized to produce and secrete FIX protein for > 5 months. We conclude that currently available adenoviral vectors have serious limitations for use for long-term gene therapy.

Adenoviridae↗
PubMed2000

Adeno-associated virus (AAV)-3-based vectors transduce haematopoietic cells not susceptible to transduction with AAV-2-based vectors.

Although adeno-associated virus (AAV)-2 has a broad tissue-host range and can transduce a wide variety of tissue types, some cells, such as erythro-megakaryoblastoid cells, are non-permissive and appear to lack the AAV-2 receptor. However, limited studies have been reported with the related dependovirus AAV-3. We have previously cloned this virus, characterized its genome and produced an infectious clone. In this study, the gene for green fluorescent protein (GFP) was inserted into AAV-2- and AAV-3-based plasmids and recombinant viruses were produced. These viruses were then used to transduce haematopoietic cells and the transduction efficiencies were compared. In contrast to recombinant (r) AAV-2, rAAV-3 successfully transduced erythroid and megakaryoblastoid cells, although rAAV-2 was superior in transduction of lymphocyte-derived cell lines. Recently, it was reported that heparan sulphate can act as a receptor of AAV-2. The infectivity of rAAV-2 and rAAV-3 was tested with mutant cell lines of Chinese hamster ovary cells that were defective for heparin or heparan sulphate expression on the cell surface. There was no correlation between the ability of rAAV-2 or rAAV-3 to infect cells and the cell surface expression of heparan sulphate and, although heparin blocked both rAAV-2 and rAAV-3 transduction, the ID(50) of rAAV-3 was higher than that of rAAV-2. In addition, virus-binding overlay assays indicated that AAV-2 and AAV-3 bound different membrane proteins. These results suggest not only that there are different cellular receptors for AAV-2 and AAV-3, but that rAAV-3 vectors may be preferred for transduction of some haematopoietic cell types.

Animals↗
PubMed1997

Construction of the temperature-sensitive vectors pLUCH80 and pLUCH88 for delivery of Tn917::NotI/SmaI and use of these vectors to derive a circular map of Listeria monocytogenes Scott A, a serotype 4b isolate.

A physical map of Listeria monocytogenes Scott A was generated by the pulsed-field technique of contour-clamped-homogeneous-electric-field (CHEF) electrophoresis. The circular genome of this serotype 4b strain contains 12 AscI fragments (38 to 790 kb), 5 NotI fragments (55 to 1,400 kb), 3 SrfI fragments (110, 1,110, and 2,000 kb), and 2 SfiI fragments (1,320 and 1,920 kb). Summation of individually sized fragments derived by digestion of Scott A genomic DNA with each of these four enzymes provided an average estimated genome length of 3,210 +/- 60 kb. Efforts to assemble the macrorestriction map benefited greatly from the construction and use of pLUCH80 and pLUCH88, temperature-sensitive vectors for delivering transposon Tn917::NotI/SmaI to the chromosome of Scott A. As another component of this study, the positions of four known virulence genes (inlA, mpl, hly, and prf) and three L. monocytogenes-specific sequences (lisM44, lisM51, and lisM52) were localized on the physical map of Scott A by hybridization. Probes prepared from lisM44, lisM51, and the four virulence genes hybridized within a cluster on a 150-kb fragment of the Scott A genome that overlaps part of the NotI-B and AscI-D fragments. The lisM52 probe hybridized with the AscI-F2 (120-kb) fragment of Scott A, which is separated from the NotI-B-AscI-D region by about 300 kb. These results established the first physical and genetic map of a serotype 4b strain of L. monocytogenes and provided further insight on this important food-borne pathogen at the genome level.

Base Sequence↗
PubMed2001

Molecular aspects of parasite-vector and vector-host interactions in leishmaniasis.

Leishmania-sand fly interactions are reviewed in the context of the potential barriers to the complete development of the parasite that exist within the midgut environment of phlebotomine flies and the molecular adaptations that the parasite has evolved that permit the development of transmissible infections to proceed. Cell surface and secreted phosphoglycans protect the parasite from the proteolytic activities of the blood-fed midgut, mediate attachment to the gut wall in order to maintain infection during excretion of the bloodmeal, and contribute to the formation of a biological plug in the anterior gut that may promote transmission by bite. The importance of vector saliva in modulating the host response to transmitted parasites is also reviewed.

Animals↗
PubMed2003

Experimental studies on the transmission of Onchocerca volvulus by its vector in the Sanaga valley (Cameroon): Simulium squamosum B. Intake of microfilariae and their migration to the haemocoel of the vector.

As large-scale ivermectin distribution is becoming the mainstay of onchocerciasis control in Africa, the issue of its impact on local transmission is increasing in importance. The vector competence of Simulium squamosum B in the severe focus of the Sanaga valley, Cameroon, was therefore investigated, by feeding 1320 flies on 14 carriers of Onchocerca volvulus microfilariae (mff). The results enabled the relationships between skin microfilarial load, microfilarial intake by the flies, the proportion and mean number of ingested mff that succeed in reaching the fly's haemocoel, and the frequency distribution of the ingested mff to be described, as functions of time post-engorgement (p.e.) and parasite density (while taking account of possible measurement error in the predictor variable). The proportion of flies with haemocoelic mff and the mean number of mff/fly increased up to 3 h p.e. The proportion of flies with ingested mff was non-linearly related to mean intake, via the negative-binomial distribution, with the overdispersion parameter k best described as an increasing (power) function of the mean. Approximately one in every three ingested mff escaped imprisonment by the peritrophic matrix, irrespective of the skin microfilarial load or the intake of mff. The relationship between successful and input mff is nearly linear (indicating proportionality) in S. squamosum B. These results are compared with those from O. volvulus-S. damnosum s.l. combinations in other West African foci.

Adolescent↗
PubMed2004

Malaria vectors in the Bioko Island (Equatorial Guinea): estimation of vector dynamics and transmission intensities.

The current study was performed on the Bioko Island (Equatorial Guinea) with the aim of establishing a rapid assessment technique for mapping malaria risk and measuring vector densities. Human bait collection, tent traps, light traps, indoor resting collection, and window exit traps were used to collect Anopheles gambiae s.s. and Anopheles funestus, the two anopheline species involved in malaria transmission in this island. Capture data were used to compare differences in the behavior and vectorial capacity of An. gambiae s.s. and An. funestus. Differences in the two species of mosquitoes were found in relation to the season and trapping methods used. Entomological inoculation rates (EIR) for Plasmodium falciparum were calculated using a polymerase chain reaction (PCR) test with individual anopheline mosquitoes from human bait collections in two villages during the dry and rainy seasons. P. falciparum sporozoites were detected from both dissected heads/thorax and abdomens of both species.

Animals↗
PubMed2002

RNAi and viral vectors as useful tools in the functional genomics of plants. Construction of BMV-based vectors for RNA delivery into plant cells.

The sequencing of several complete genomes and the development of a DNA microarray technology are among the most important achievements of molecular biology. They gave the proper grounds for the development of modern functional genomics. However, there is one additional condition which needs to be satisfied to truely enable the study of how a genome works: a suitable method of selectively inducing and silencing the expression of each individual gene. The methods used so far have usually only permitted the influencing of gene expression through genetic manipulations at the DNA level (genetically modified plants). The discovery of RNA interference (RNAi) opens up completely new possibilities of research on the functioning of particular plant genes, without the necessity of altering the genome structure. In this case, interference takes place at the transcript level. Thus, at any given moment during plant development, the expression of a specific gene (or several genes) can be inhibited, even if it is important for the survival of the organism under study. To this end, a double-stranded RNA inducing the RNAi phenomenon has to be delivered into the plant cell. Here we describe the construction of four brome mosaic virus-based vectors, which, as our preliminary data indicate, can be used to transfer RNA into barley cells.

Bromovirus↗
PubMed2000

[Insecticide sensitivity in malaria vectors in high altitude Madagascar after five years of vector control].

In 1991, Anopheles gambiae and Anopheles funestus, the main malaria vectors in the Highlands of Madagascar, were reported to be fully susceptible to DDT; nevertheless a slight decrease in the susceptibility levels was recorded when compared with previous assays carried out in 1962. From 1993 to 1997, five cycles of indoor residual spraying have been carried out in the Highlands: a total of 1,482,000 kg of 70% wp DDT have been used for the treatment of houses and animal shelters. From 1996 to 1999, adult mosquito susceptibility tests to DDT and to some pyrethroids (lambdacyalothrine, deltamethrine, permethrine and cyfluthrine) have been carried out on samples collected in 20 areas of the Highlands. Bioassays were carried out following the WHO standard method. All tested populations of An. funestus showed a full susceptibility to DDT. An. gambiae showed a widespread decrease in the susceptibility to DDT, particularly marked in the region of the capital city Antananarivo. Both species were susceptible to pyrethroids.

Adult↗
PubMed1989

Vectors of Japanese encephalitis virus (JEV): species complexes of the vectors.

The vectors of JEV are Cx. tritaeniorhynchus, Cx. vishnui, Cx. pseudovishnui, Cx. gelidus, Cx. fuscocephala, Cx. quinquefasciatus, Cx. pipiens pallens, Cx. bitaeniorhynchus, Cx. annulirostris, Aedes togoi, Ae. japonicus, Ae. vexans nipponii, Anopheles annularis and An. vagus. Cx. tritaeniorhynchus is in the tritaeniorhynchus complex, breeds in rice fields, ground pools in vast areas. Two types of mating behavior, eurygamy and moderate stenogamy were detected. In the case of the eurygamy type, the mosquitoes were from Southern Thailand and hilly areas near Kanchanaburi, Thailand. Female mosquitoes are usually dark in color, the cibarial armature has rod teeth and the posterior end of the cibarial armature is bowl shaped with a typical rim. The rim of the bowl is everted. The moderate stenogamy type were mosquitoes from the plain areas such as Bangkok, Ayutthaya, Suphan Buri and Saraburi. The posterior end of the cibarial armature is bowl shaped with a stout rim. The larvae were characteristic in their siphon index, antennal index, hair O of prothoracic segment, and comb scale number and arrangement. Cx. tritaeniorhynchus summorosus from Japan, Los Banos and Luzon, Philippines, differed from Cx. tritaeniorhynchus in that on the lateral plate of the phallosome tritaeniorhynchus teeth are somewhat weakly developed and only gently curved whereas in tritaeniorhynchus summorosus they are strongly developed, considerably longer, and sharply recurved. The siphons of larvae are short, the sides parallel and the apex truncate in tritaeniorhynchus whereas in tritaeniorhynchus summorosus they are long and slender. Cx. tritaeniorhynchus var. siamensis is possibly present. Colonies have been maintained in the Department of Medical Entomology for 31 generations. The characteristics are in hair O (short, less than 20 branches, and without secondary branching and the larval siphon (short and broad where the others are long). Cx. vishnui and Cx. pseudovishnui are in the vishnui complex. Cx. quinquefasciatus and Cx. pipiens pallens are in the Cx. pipiens complex comprising: (1) Cx. pipiens; (2) Cx. quinquefasciatus Say; (3) Cx. molestus Forskal; (4) Cx. pipiens pallens; (5) Cx. australicus; (6) Cx. globocoxitus. Anopheles annularis is a species complex evidenced by two types of polytene chromosomes.

Animals↗
PubMed2002

Temperature dependence of rho and a1-meson masses and mixing of vector and axial-vector correlators.

Within a chiral model which provides a good description of the properties of rho and a1 mesons in vacuum, it is shown that, to order T2, the rho- and a1-meson masses remain constant in the chiral limit, even if at tree level they are proportional to the chiral condensate, sigma0. Numerically, the temperature dependence of the masses turns out to be small also for realistic parameter sets and high temperatures. The weak temperature dependence of the masses is consistent with the Eletsky-Ioffe mixing theorem, and traces of mixing effects can be seen in the spectral function of the vector correlator at finite temperature.

Journal Article↗
PubMed2003

Rates, polarizations, and asymmetries in charmless vector-vector B meson decays.

With a sample of approximately 89 x 10(6) B(-)B pairs collected with the BABAR detector, we perform a search for B meson decays into pairs of charmless vector mesons (phi, rho, and K*). We measure the branching fractions, determine the degree of longitudinal polarization, and search for CP violation asymmetries in the processes B+-->phiK(*+), B0-->phiK(*0), B+-->rho(0)K(*+), and B+-->rho(0)rho(+). We also set an upper limit on the branching fraction for the decay B0-->rho(0)rho(0).

Journal Article↗