Expression and accumulation of the two-domain odorant-binding protein AaegOBP45 in the ovaries of blood-fed Aedes aegypti

BACKGROUND: \ud Aedes aegypti mosquitoes are the main vectors of dengue viruses. Despite global efforts to reduce the prevalence of dengue using integrated vector management strategies, innovative alternatives are necessary to help prevent virus transmission. Detailed characterizations of Ae. aegypti genes and their products provide information about the biology of mosquitoes and may serve as foundations for the design of new vector control methods.\ud FINDINGS: \ud We studied the Ae. aegypti gene, AAEL010714, that encodes a two-domain odorant-binding protein, AaegOBP45. The predicted gene structure and sequence were validated, although single nucleotide polymorphisms were observed. Transcriptional and translational products accumulate in the ovaries of blood fed females and are not detected or are at low abundance in other tissues.\ud CONCLUSIONS: \ud We validated the Ae. aegypti AAEL010714 gene sequence and characterized the expression profile of a two-domain OBP expressed in ovaries. We propose that AaegOBP45 function as a component of the mosquito eggshell.São Paulo Research Foundation (FAPESP) 2006/00331-3CNPq 143113/2006-2NIH NIAID (AI29746

Endogenously-expressed NH2-terminus of circumsporozoite protein interferes with sporozoite invasion of mosquito salivary glands

Abstract\ud \ud Background\ud The circumsporozoite protein is the most abundant polypeptide expressed by sporozoites, the malaria parasite stage capable of infecting humans. Sporozoite invasion of mosquito salivary glands prior to transmission is likely mediated by a receptor/ligand-like interaction of the parasites with the target tissues, and the amino (NH2)-terminal portion of CSP is involved in this interaction but not the TSR region on the carboxyl (C)-terminus. Peptides based on the NH2-terminal domain could compete with the parasites for the salivary gland receptors and thus inhibit penetration.\ud \ud \ud Methods\ud Peptides based on the NH2-terminus and TSR domains of the CSP from avian or human malaria parasites, Plasmodium gallinaceum and Plasmodium falciparum, respectively, were expressed endogenously in mosquito haemolymph using a transient (Sindbis virus-mediated) or stable (piggyBac-mediated transgenesis) system.\ud \ud \ud Results\ud Transient endogenous expression of partial NH2-terminus peptide from P. falciparum CSP in P. gallinaceum-infected Aedes aegypti resulted in a reduced number of sporozoites in the salivary glands. When a transgenic approach was used to express a partial CSP NH2-terminal domain from P. gallinaceum the number of sporozoites in the salivary glands did not show a difference when compared to controls. However, a significant difference could be observed when mosquitoes with a lower infection were analysed. The same result could not be observed with mosquitoes endogenously expressing peptides based on the TSR domain from either P. gallinaceum or P. falciparum.\ud \ud \ud \ud Conclusion\ud These results support the conclusion that CSP partial NH2-terminal domain can be endogenously expressed to promote a competition for the receptor used by sporozoites to invade salivary glands, and they could be used to block this interaction and reduce parasite transmission. The same effect cannot be obtained with peptides based on the TSR domain.We thank Neuza Saraiva Fernandes and Ediane Saraiva Fernandes for technical\ud assistance and Alexandre Santos de Moura for sample sequencing. This work\ud was supported by FAPESP (Process#04/00889-9 and#00/12138-7). AAJ was\ud supported in part by an award from the National Institute of Allergy and Infec‑\ud tious Diseases (USA) (R37AI029746)

Expression and accumulation of the two-domain odorant-binding protein AaegOBP45 in the ovaries of blood-fed Aedes aegypti

Abstract Background Aedes aegypti mosquitoes are the main vectors of dengue viruses. Despite global efforts to reduce the prevalence of dengue using integrated vector management strategies, innovative alternatives are necessary to help prevent virus transmission. Detailed characterizations of Ae. aegypti genes and their products provide information about the biology of mosquitoes and may serve as foundations for the design of new vector control methods. Findings We studied the Ae. aegypti gene, AAEL010714, that encodes a two-domain odorant-binding protein, AaegOBP45. The predicted gene structure and sequence were validated, although single nucleotide polymorphisms were observed. Transcriptional and translational products accumulate in the ovaries of blood fed females and are not detected or are at low abundance in other tissues. Conclusions We validated the Ae. aegypti AAEL010714 gene sequence and characterized the expression profile of a two-domain OBP expressed in ovaries. We propose that AaegOBP45 function as a component of the mosquito eggshell

Influence of criminal activity of sentenced women and of their imprisonment during an execution of punishment on perception of guilt and meaningfulness of life

The doctoral thesis is engaged in the question of guilt, compassion - repentance, forgiveness, responsibility and meaningfulness of existence in the sentenced women's lives at the beginning and during their execution of punishment. Guilt is presented from Christian, philosophical- psychological point of view and also from the perspective of law. Guilt is usually prosecuted by punishment. The next chapter analyzes what kind of punishment is adequate to what guilt, which punishment is positive and which one is negative. The next chapter is devoted to a term of compassion-or repentance- as an inevitable part of the forgiving process. An author perceives forgiveness towards self and others as a necessary part of "the healing process", towards which the work of specialists in execution of punishment should be directed. Forgiveness is unthinkable without acceptance of own responsibility for one's decisions and deeds. Perception of own meaningfulness is connected with all mentioned topics. The last chapter in the theoretical part of the paper is devoted to the subject of meaning of life and loss of its meaning. The practical part focuses on the group of women sentenced for a violent crime and on the group of women sentenced for property crime. Each group is further divided according to length of their imprisonment..

Integrated proteomic and transcriptomic analysis of the Aedes aegypti eggshell

BACKGROUND: Mosquito eggshells show remarkable diversity in physical properties and structure consistent with adaptations to the wide variety of environments exploited by these insects. We applied proteomic, transcriptomic, and hybridization in situ techniques to identify gene products and pathways that participate in the assembly of the Aedes aegypti eggshell. Aedes aegypti population density is low during cold and dry seasons and increases immediately after rainfall. The survival of embryos through unfavorable periods is a key factor in the persistence of their populations. The work described here supports integrated vector control approaches that target eggshell formation and result in Ae. aegypti drought-intolerant phenotypes for public health initiatives directed to reduce mosquito-borne diseases. RESULTS: A total of 130 proteins were identified from the combined mass spectrometric analyses of eggshell preparations. CONCLUSIONS: Classification of proteins according to their known and putative functions revealed the complexity of the eggshell structure. Three novel Ae. aegypti vitelline membrane proteins were discovered. Odorant-binding and cysteine-rich proteins that may be structural components of the eggshell were identified. Enzymes with peroxidase, laccase and phenoloxidase activities also were identified, and their likely involvements in cross-linking reactions that stabilize the eggshell structure are discussed

Transcriptome Sequencing and Developmental Regulation of Gene Expression in <i>Anopheles aquasalis</i>

<div><p>Background</p><p><i>Anopheles aquasalis</i> is a major malaria vector in coastal areas of South and Central America where it breeds preferentially in brackish water. This species is very susceptible to <i>Plasmodium vivax</i> and it has been already incriminated as responsible vector in malaria outbreaks. There has been no high-throughput investigation into the sequencing of <i>An. aquasalis</i> genes, transcripts and proteins despite its epidemiological relevance. Here we describe the sequencing, assembly and annotation of the <i>An. aquasalis</i> transcriptome.</p><p>Methodology/Principal Findings</p><p>A total of 419 thousand cDNA sequence reads, encompassing 164 million nucleotides, were assembled in 7544 contigs of ≥2 sequences, and 1999 singletons. The majority of the <i>An. aquasalis</i> transcripts encode proteins with their closest counterparts in another neotropical malaria vector, <i>An. darlingi</i>. Several analyses in different protein databases were used to annotate and predict the putative functions of the deduced <i>An. aquasalis</i> proteins. Larval and adult-specific transcripts were represented by 121 and 424 contig sequences, respectively. Fifty-one transcripts were only detected in blood-fed females. The data also reveal a list of transcripts up- or down-regulated in adult females after a blood meal. Transcripts associated with immunity, signaling networks and blood feeding and digestion are discussed.</p><p>Conclusions/Significance</p><p>This study represents the first large-scale effort to sequence the transcriptome of <i>An. aquasalis</i>. It provides valuable information that will facilitate studies on the biology of this species and may lead to novel strategies to reduce malaria transmission on the South American continent. The <i>An. aquasalis</i> transcriptome is accessible at <a href="http://exon.niaid.nih.gov/transcriptome/An_aquasalis/Anaquexcel.xlsx" target="_blank">http://exon.niaid.nih.gov/transcriptome/An_aquasalis/Anaquexcel.xlsx</a>.</p></div

Distribution of the best matches of all <i>An. aquasalis</i> predicted proteins by organisms: Andar- <i>An. darlingi</i>; Angam- <i>An. gambiae</i>; Aeaeg, <i>Ae. aegypti</i>; Cuqui, <i>Cu. quinquefasciatus</i>; Drmel, <i>D. melanogaster</i>; Other <i>Anopheles</i> species; Other insects- not of the <i>Anopheles</i> genus; Other- non-insect organisms.

<p>Distribution of the best matches of all <i>An. aquasalis</i> predicted proteins by organisms: Andar- <i>An. darlingi</i>; Angam- <i>An. gambiae</i>; Aeaeg, <i>Ae. aegypti</i>; Cuqui, <i>Cu. quinquefasciatus</i>; Drmel, <i>D. melanogaster</i>; Other <i>Anopheles</i> species; Other insects- not of the <i>Anopheles</i> genus; Other- non-insect organisms.</p

Number of sequences composing the assembled <i>An. aquasalis</i> contigs.

<p>A total of 7544 contigs were assembled from ≥2 sequences. The number of sequences that compose each contig varies from 2 to 5,207, with an average of 35 sequences per contig. Forty-three percent of the assembled contigs contained 10 or more sequences.</p