1,721,000 research outputs found
Thrombospondin related anonymous protein (TRAP) of Plasmodium falciparum in parasite-host cell interactions.
No full textThrombospondin related anonymous protein (TRAP) of Plasmodium falciparum is characterized by the presence of an amino acid motif based on the sequence Trp-Ser-Pro-Cys-Ser-Val-Thr-Cys-Gly (WSPCSVTCG) that is found in a growing family of proteins. The sequence WSPCSVTCG is considered to confer sulpho-galactosyl-cerebroside (sulphatide) binding properties to antistasin, TSP, CS protein and properdin. The observation that TRAP is localized both on the micronemes and on the surface of P. falciparum sporozoites would suggest a role played by TRAP, and its putative sulphated glycoconjugates binding motif, in the recognition and/or entry of hepatocytes by the sporozoite. Our results indicated that TRAP constructs, expressed in E. coli, bind to sulpho-galactosyl-cerebrosides (sulphatides) and to the surface of HepG2 cells using the conserved amino acid motif WSPCSVTCG. Antisera raised against TRAP constructs inhibited sporozoite invasion of HepG2 cells thus suggesting, thus, that TRAP may be one of the parasite-encoded molecules implicated in the sporozoite invasion of hepatocytes. Moreover, the possibility that TRAP antibodies may be relevant in malaria immunity is supported by the results obtained in a prospective study conducted in a malaria endemic area. In adolescents, the presence of TRAP antibodies, before malaria transmission, correlated positively with the control of parasite density
A method for collecting large quantities of Cryptosporidium parasites
No full textThe intestinal mucoso may be regarded as a potential and abundant source of Cryptosporidium parvurn parasites from which all developmental stages might be collected. If intracellular stages could be recovered from the brush border, many of the limitations concerned with the use of oocysts and in vitro cultures may be overcome. Hans-Michael Muller, Lorella Ranucci, Edoordo Pozio and Andrea Crisanti discuss here how this can be done
Temporal and spatial expression of serine protease genes in Anopheles gambiae.
No full textSerine proteases play a crucial role during the digestion of the blood meal in the mosquito gut. The isolation and the analysis of the genomic organisation of the corresponding genes may lead to the characterization of gut-specific, inducible promoters, suitable for the expression of anti-parasitic agents in the gut of transgenic mosquitoes. We report here on the identification of a trypsin and a chymotrypsin gene family of Anopheles gambiae. Following a blood meal, the transcription of all members of the two identified gene clusters, seven trypsin genes (Antryp1-7) and two chymotrypsin genes (Anchym1-2), is induced. Recombinant Antryp1 and Antryp2, expressed in E. coli, were both active in vitro against blood proteins. Moreover, mouse sera raised against Antryp1, Anchym1 and Anchym2 recognized the corresponding proteases among the proteins of a lysate prepared from dissected guts of An. gambiae mosquitoes
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Constitutive and blood meal-induced trypsins of Anopheles gambiae
No full textTrypsin genes in Anopheles gambiae are arranged as a tightly clustered gene family consisting of seven related coding sequences, devoid of introns. The two blood meal-inducible members of this family, Antryp1 and Antryp2, were shown to play a crucial role in the breakdown of the blood meal constituents. The role of Antryp3,4,5,6, and Antryp7 in the process of blood meal digestion remains to be elucidated. We have examined the localization and the expression patterns of these trypsins as well as the functional interactions in blood meal digestion between trypsins and other gut-specific proteases. Northern blot and RT-PCR analysis indicated that the genes Antryp3,4,5,6, and Antryp7 are all constitutively expressed in unfed female mosquitoes. Soon after blood feeding the mRNA of these trypsin genes became undetectable and appeared again at the end of the gonotrophic cycle. The blood meal-inducible trypsin Antryp1 was also constitutively expressed at low level in the gut of adult female mosquitoes. This trypsin was the only member of this gene family to be expressed in the gut of male and female pupae. By using antisera that specifically recognized recombinant Antryp4 we were able to show that the corresponding protein in Anopheles is synthesized and stored in the gut epithelium of unfed females as zymogen. Secretion and activation of this trypsin was shown to occur in the midgut lumen immediately after fluid ingestion and independently of the protein content of the meal. Recombinant trypsins expressed in Escherichia coli, with the exception of Antryp5 and Antryp6, were able to activate in vitro recombinant A. gambiae chymotrypsinogen, thus suggesting that blood meal ingestion is able to trigger a cascade of events leading to the activation of several proteases
Conserved function of anopheles gambiae midgut-specific promoters in the fruitfly.
No full textControl of malaria by a methodology that would permit the effective blockage of the Anopheles gambiae midgut wall penetration by Plasmodium parasites requires a detailed understanding of both the physiology of the mosquito's digestion, and of the interactions between the parasite and its host. We have transformed Drosophila melanogaster with several constructs that allow the study of the promoter region of two of the major late trypsin genes of A. gambiae. Using several deletions, we have identified, for both genes, small genomic segments that are sufficient to confer tissue specificity to the promoter in a species that is far away in evolution from the mosquito. This will allow further studies that will enable both the understanding of the blood meal digestion, and may potentially be useful for the design of anti-plasmodial constructs at a later stage
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Molecular cloning and expression of a 70-kilodalton heat shock protein of Candida albicans.
No full textBy screening an expression library of the yeast form of Candida albicans with a serum directed against whole fungal cells, a cDNA (2,325 bp) encoding a stress protein of C. albicans was cloned and sequenced. The cloned sequence (CaRLV130) identified a single open reading frame with a length of 1,968 bp coding for a protein containing 656 amino acid residues (70 kDa). The deduced amino acid sequence was 84% similar to the sequence of the Saccharomyces cerevisiae SSA1 gene, which encodes one member of the 70-kDa heat shock protein (Hsp70) family. The relevant gene (C. albicans HSP70 gene [CaHSP70]) was localized on the highest-M(r) (R1; approximately 3.8 Mb) chromosome of C. albicans as determined by pulse-field electrophoresis. CaHSP70 was expressed after heat shock, as demonstrated by Northern (RNA) blotting and reverse transcriptase-PCR with specific pairs of oligonucleotide sequences and gene probes. A recombinant protein was obtained in Escherichia coli after cloning of the full coding sequence into the BamHI site of the pDS56/RBSII6xhisE- plasmid and purification by nickel chelate affinity chromatography. The recombinant protein (6xhis-CaHsp70) was efficiently recognized in immunoblots by a monoclonal antibody directed against a common epitope of eukaryotic Hsp70 proteins, as well as by sera from normal human subjects. Moreover, immune mouse sera against the purified recombinant protein recognized native, heat-inducible constituents with sizes of around 70 kDa in whole-cell protein extracts of C. albicans. Overall, our data demonstrate that CaHSP70 encodes one member of a family of proteins (Hsp70) which usually represent highly conserved immunodominant antigens of infectious agent
Development of the human immune response against the major surface protein (gp190) of Plasmodium falciparum.
No full textThe 190-kilodalton glycoprotein (gp190) of Plasmodium falciparum, the precursor of the major surface proteins of merozoites, is considered a promising candidate for a blood stage malaria vaccine. DNA sequences specific for the gp190 of the two isolates K1 and MAD20 were subcloned and expressed in Escherichia coli. The panel of fusion proteins obtained represents about 80% of the polymorphic sequences observed so far within various isolates of P. falciparum. Sera from individuals living in a malaria-endemic area of West Africa were tested in immunoblots against the gp190 fusion proteins, and antibody reactivity was mapped to defined regions of the gp190. Depending on the age of the individual and on the presence of parasites in the blood, distinct regions of gp190 were differentially recognized by the respective antibodies. Similarly, the analysis of sera from German patients with acute malaria revealed a distinct pattern. When grouped according to age and to parasitemia, the reactivity of the sera of people living in malaria-endemic areas may indicate a correlation between certain gp190 regions and protective immune response
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