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Biomedical subjects

V Robert

Publications and source records attributed to V Robert.

At least 127 records · Page 7Linked to original sources

Characterization of monoclonal antibodies identifying type and strain-specific epitopes of human immunodeficiency virus type 1.

Several hybridoma cell lines were raised against the highly cytopathic Zaïrian isolate of Human Immunodeficiency Virus (HIV), HIV1-NDK. The specificity of the secreted monoclonal antibodies (mAb) was demonstrated by immunoblotting, radioimmunoprecipitation and immunofluorescence. Two hybridoma cell lines secreted mAb reacting with independent epitopes of the NDK p17 capsid protein and its precursors. One, RL16.24.5, is specific for the NDK isolate whereas the other, RL16.45.1, along with anti-p25 RL16.30.1 mAb, bind all HIV1 isolates but not HIV2. Together with the previously described mAb RL4.72.1 those reagents define lentivirus subfamily (HIV1, HIV2, SIV) type/subtype (HIV1) and strain (HIV1-NDK) specific epitopes expressed on HIV1-NDK core proteins. The last mAb RL16.76.1 binds the env gene products gp160 and gp120.

Animals↗

Expression of an immunogenic region of HIV by a filamentous bacteriophage vector.

Vectors derived from the Escherichia coli filamentous phage, fd-tet, expressing parts of the human immunodeficiency virus (HIV) gag genes were constructed and analyzed. The immunoreactive domain of HIV Gag antigens was produced in the form of a fusion protein, with a phage minor coat protein, called protein III, playing an important role in phage infectivity. A micropanning procedure, utilizing the strong affinity of biotinylated antibody to streptavidin, was applied for the selection of clones. A simple preparation procedure consisting of polyethyleneglycol precipitation of the recombinant phage from the E. coli supernatant allowed us to detect HIV antigens by enzyme-linked immunosorbent assay (ELISA). Cloned FUSE-gag, as isolated using anti-Gag RL4.72.1 monoclonal antibody (mAb), contained a nucleotide sequence coding for 91 amino acids of HIV Gag p24. It specifically reacted with the mAb in the ELISA. Construction of the mAb-selectable phages permitted localization of epitopes for mAb. Infectivity of the phage clone was specifically neutralized by the anti-HIV mAb. Immunoelectroblotting analysis of recombinant phages revealed the presence of an about 65-kDa band reacting with anti-HIV mAb. This Mr corresponded to the size of the fused form of the FUSE 1 protein III. Human sera from HIV-infected and uninfected individuals reacted with recombinant protein III, as well as the original form of protein III.

Amino Acid Sequence↗

Photoduplication and fluorescence measurement in polyacrylamide gel electrophoresis.

A simple method of photoduplication of gel electrophoresis, visualized with fluorescent reagents, is described. The procedure is convenient and rapid and requires no camera or expensive equipment. Using electrophoresis duplicating paper (Kodak), positive prints suitable for documentation or publication may be obtained. With usual photographic paper, negative prints may be obtained, allowing reliable measurement by scanning. The technique may be applied to protein or nucleic acid electrophoresis.

Copying Processes↗

Effects of hydroxyl radicals on ATPase and protein structure of myofibrils from rat heart.

Free oxygenated radicals frequently are involved in cardiac arrhythmias and contractility disorders during postischemic reperfusion. The aim of this study was to determine the effects of hydroxyl radicals (.OH) in vitro on myofibrillar Ca-adenosinetriphosphatase (ATPase), on the redox state of thiol groups and the electrophoretic pattern of myofibrillar proteins from rat heart. Myofibrils were treated up to 60 min by .OH generated with 0.3 mM H2O2 and 0.1 mM Fe2+. After a 60-min treatment with .OH, the measurement of thiol groups failed to show any oxidation. On the contrary, ATPase activity and electrophoretic pattern were affected dramatically by treatment with .OH. For all Ca2+ concentrations, ATPase was increased after treatment with .OH, but ATPase activation when Ca2+ rose from pCa 8 to pCa 4.5 was only 92% after 30 min of incubation rather than 226% for untreated myofibrils. The electrophoretic analysis of myofibrillar proteins showed a decrease in myosin heavy chain and formation of aggregates in treated myofibrils. All of these effects were reduced when incubation was performed in the presence of mannitol, a specific scavenger of .OH. No effect was observed with 0.1 mM Fe2+ alone or with 0.3 mM H2O2. The action of .OH was very fast to the extent that the effects were observed after only 15 s of incubation. The results reported in the present study may be related to the impaired relaxation and contracture described in vivo within the first minutes of a postischemic reperfusion and before any change in calcium homeostasis.

Animals↗

Plasmodium falciparum and P. malariae epidemiology in a West African village.

Transmission of Plasmodium falciparum and P. malariae was studied in a village in Burkina Faso. Consecutive captures of mosquitos were organized twice a month over a year and the species of the mosquitos identified. Also, the prevalences and densities of Plasmodium spp. were determined every 2 months in a sample of children who lived in the village. Anopheles gambiae, A. funestus, and A. nili were the local vectors, but only the first two played a predominant role in both P. falciparum and P. malariae transmission. The parasitological sporozoite index (SI) was 4.48% for A. gambiae and 4.22% for A. funestus. The immunological SIs were higher: 5.82% of A. gambiae were infected with P. falciparum and only 0.16% with P. malariae; the corresponding proportions for A. funestus were 6.45% and 0.41%. Transmission of Plasmodium spp. by A. gambiae was important during the rainy season (July-October) and by A. funestus at the beginning of the dry season (September-November). Each child in the study village could receive about 396 P. falciparum-infected bites per year but only 22 of P. malariae. The P. falciparum parasite indices were maximum during the middle of the rainy season (August), while those for P. malariae reached a peak during the dry season (February).

Adolescent↗

Influence of deltamethrin treatment of bed nets on malaria transmission in the Kou valley, Burkina Faso.

A 3-year entomological study was carried out on the transmission of malaria in a village of 900 inhabitants in a rice-growing area of Burkina Faso. In the study area inhabitants use bed nets to protect themselves from mosquito bites. In the first year of the study, baseline data were collected; in the second year, the village was divided in two parts and all the bed nets in the southern part were sprayed with deltamethrin (25 mg/m2); and in the third year, all the bed nets in both parts of the village were sprayed. The inoculation rate was estimated by hand collection of mosquitos on human volunteers who were not protected by bed nets. The overall inoculation rate in the first year was 55 infected bites per person and was higher in the southern than in the northern part of the village. During the second year the rate increased to 70 bites per person on average (but was slightly lower than this in the southern part of the village). During the third year, the inoculation rate fell to three infected bites per year, i.e., a reduction of 94% compared with the first year. This reduction arose primarily because of a marked decrease in the sporozoitic index and a lower density of vectors. Thus, use of pyrethroid-impregnated bed nets by all members of the community appears to be a major tool in preventing transmission of malaria.

Animals↗

[The impact of impregnated mosquito nets on prevalence and morbidity related to malaria in sub-Saharan africa].

Insecticide treated bed nets (permethrin, deltamethrin and lambda cyalothrin) were used for malaria control in The Gambia, Burkina Faso and Tanzania where Anopheles gambiae (and An.funestus in Burkina Faso) is the main vector. Treated mosquito nets are efficient when used on a large scale and not on an individual level. Such a large scale use, acting on longevity and infectivity of vectors, always induced a decrease of malaria transmission by more than 90%. Treated bed nets had no significant effect on the overall parasite rate, showing that malaria transmission was not stopped. But it was usually found that there was a significantly smaller number of children with parasitemia higher than a critical threshold, a sensitive parameter of malaria morbidity. Indeed, in the three situations studied, malaria morbidity (fever + high parasitemia greater than critical level in the concerned area) has generally shown a similar drop of 60%. These data demonstrate that treated bed nets are useful to reduce transmission and morbidity. Advised as a new way for reduction of nuisance, treated bed nets were always welcomed by the populations and this method may be considered as a complementary weapon in public health.

Adolescent↗

[The effect of 2 chloroquine-based drug strategies (prevention and therapy of febrile cases] on malaria transmission].

A 3 years study was decided in 12 villages of the South-West Burkina Faso to compare the chemoprophylaxis and the chemotherapy of febrile cases as potential malaria control strategies. During the first year pretreatment data were collected. During the two following years a programme carried out (I) prophylaxis (10 mg chloroquine/kg body weight) was given weekly to all children under 14 years old in 5 villages, and (II) therapy (10 mg chloroquine/kg body weight) was given in a single dose to all febrile cases in 7 other villages. Chloroquine tablets were distributed by health workers belonging to the community. Both prophylaxis and therapy reduced the gametocyte rate in children (2-9 years) respectively of 63% and 45%. The analysis of the evolution after the first year of the sporozoite rate of anopheline was made difficult by concomitant natural variations of mosquitoes longevity and by mosquitoes displacements. Significant variations of sporozoite rate can be explained by natural variations of mosquitoes longevity. But data from the rice field villages support evidence that reduction of the pool of parasite infective for vectors induced the decrease of Anopheles gambiae s. l. sporozoite rate. Therefore our results reflect a trend more than a strict reduction of malaria transmission.

Adolescent↗

Study of the distribution of circumsporozoite antigen in Anopheles gambiae infected with Plasmodium falciparum, using the enzyme-linked immunosorbent assay.

Anopheles gambiae, experimentally infected with Plasmodium falciparum, were dissected 14 days later for microscopical detection of sporozoites and oocysts. The head, salivary glands, thorax, midgut, legs, ovaries, Malpighian tubules, the remainder of the abdominal tissues and the dissection fluid of each mosquito were examined by a two-site enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of circumsporozoite antigen (CS ag). 19 mosquitoes had CS ag in at least one of the specimens examined. Very large individual variability was observed in the presence and/or quantity of CS ag in the various parts. 7 mosquitoes were ELISA-positive in all 9 specimens; the salivary glands and thorax contained most CS ag, whereas the Malpighian tubules and ovaries contained the least; all the thoraces contained CS ag, even that of one mosquito of which the salivary glands lacked both sporozoites and CS ag; of 17 ELISA-positive salivary glands, 15 were found to contain sporozoites. The existence of free antigen associated with sporozoites, and the limitations of the ELISA technique in demonstrating the infectivity of a malaria vector, are discussed.

Animals↗

[An ecologic study of adult and larval Culicidae in a rice field of Kou Valley, Burkino Faso].

An entomological survey based on collections of human bait mosquitoes and of mosquito larvae was carried out through one year in a rice-field, the Kou Valley of southwest Burkina Faso. Each year, in this irrigated rice field there are two crops of rice, one during the dry season and another during the wet one. Between the two rice cycles irrigation is interrupted and the rice field becomes dry. The rice crop cycle moves through several stages, all of which have positive or negative effects on the development of the larvae of most abundant mosquitoes species. Rice cultivation does not explain however entirely the ecology of mosquito populations. Two main limiting factors for mosquito development are highlighted: the season with two periods (1. dry, 2. wet) and the rice cultivation with three periods (1. start of the flooding, growing rice transplantation and tillering, 2. rice heading and flowering, 3. rice maturation and harvest). The development of a majority of mosquito species depends strictly on rice growth: (i) Anopheles gambiae s.1. uses the whole rice field during the first period of rice cultivation until growing rice protects the larval breeding places from solar radiation. It is the most frequent mosquito (53% of the man-biting mosquitoes caught). 5% of its larvae are parasitized with a fungus Coelomomyces sp., (ii) A. pharoensis has its maximum density during the second period of rice cultivation, as Culex gr. decens; both seem to succeed better in the dry season, contrary to all other anophelines, (iii) A. coustani profits by the third period when the pH of the breeding places becomes basic.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗