Identification and characterization of the fibrinogen-like domain of fibrinogen-related proteins in the mosquito, Anopheles gambiae, and the fruitfly, Drosophila melanogaster, genomes

BACKGROUND: The fibrinogen-like (FBG) domain, which consists of approximately 200 amino acid residues, has high sequence similarity to the C-terminal halves of fibrinogen β and γ chains. Fibrinogen-related proteins (FREPs), which contain FBG domains in their C-terminal region, are found universally in vertebrates and invertebrates. In invertebrates, FREPs are involved in immune responses and other aspects of physiology. To understand the complexity of this family in insects, we analyzed FREPs in the mosquito genome and made comparisons to FREPs in the fruitfly genome. RESULTS: By using the genome data of the mosquito, Anopheles gambiae, 53 FREPs were identified, whereas only 20 members were found in the Drosophila melanogaster genome. Using sequence profile analysis, we found that FBG domains have high sequence similarity and are highly conserved throughout the FBG domain region. By secondary structure analysis and comparison, the FBG domains of FREPs are predicted to function in recognition of carbohydrates and their derivatives on the surface of microorganisms in innate immunity. CONCLUSION: Detailed sequence and structural analysis discloses that the FREP family contains FBG domains that have high sequence similarity in the A. gambiae genome. Expansion of the FREP family in mosquitoes during evolutionary history is mainly accounted for by a major expansion of the FBG domain architecture. The characterization of the FBG domains in the FREP family is likely to aid in the experimental analysis of the ability of mosquitoes to recognize parasites in innate immunity and physiologies associated with blood feeding

Epidemiology and Control of a Head Louse Outbreak in Ames, Iowa, 1976

At the onset of school in 1976, head lice were recognized by teachers and school nurses in the public and parochial schools in Ames, Iowa. One or more cases were identified from the 9 elementary schools and 3 secondary schools in the community. The greatest prevalence of louse infestations occurred in 2 elementary schools (1 parochial) where 16% and 14%, respectively, of the students were infested. A program was established to screen children and exclude infested individuals from class until they were effectively treated. The community was surveyed to determine the extent of infestation in each household from which at least 1 member was attending school and had a confirmed louse infestation. Data were tabulated on the incidence of louse infestations by school, grade, sex, hair color, hair length, number of persons living in homes where louse infestations were recognized and incomes of families that had infested members. Factors that contributed to louse transmission and an evaluation of an intensive community-wide control program are discussed

Distribution of Brugia malayi larvae and DNA in vector and non-vector mosquitoes: implications for molecular diagnostics

<p>Abstract</p> <p>Background</p> <p>The purpose of this study was to extend prior studies of molecular detection of <it>Brugia malayi </it>DNA in vector (<it>Aedes aegypti- </it>Liverpool) and non-vector (<it>Culex pipiens</it>) mosquitoes at different times after ingestion of infected blood.</p> <p>Results</p> <p>Parasite DNA was detected over a two week time course in 96% of pooled thoraces of vector mosquitoes. In contrast, parasite DNA was detected in only 24% of thorax pools from non-vectors; parasite DNA was detected in 56% of midgut pools and 47% of abdomen pools from non-vectors. Parasite DNA was detected in vectors in the head immediately after the blood meal and after 14 days. Parasite DNA was also detected in feces and excreta of the vector and non-vector mosquitoes which could potentially confound results obtained with field samples. However, co-housing experiments failed to demonstrate transfer of parasite DNA from infected to non-infected mosquitoes. Parasites were also visualized in mosquito tissues by immunohistololgy using an antibody to the recombinant filarial antigen Bm14. Parasite larvae were detected consistently after mf ingestion in <it>Ae. aegypti- </it>Liverpool. Infectious L3s were seen in the head, thorax and abdomen of vector mosquitoes 14 days after Mf ingestion. In contrast, parasites were only detected by histology shortly after the blood meal in <it>Cx. pipiens</it>, and these were not labeled by the antibody.</p> <p>Conclusion</p> <p>This study provides new information on the distribution of filarial parasites and parasite DNA in vector and non-vector mosquitoes. This information should be useful for those involved in designing and interpreting molecular xenomonitoring studies.</p

Dual RNA-seq of parasite and host reveals gene expression dynamics during filarial worm–mosquito interactions

Parasite biology, by its very nature, cannot be understood without integrating it with that of the host, nor can the host response be adequately explained without considering the activity of the parasite. However, due to experimental limitations, molecular studies of parasite-host systems have been predominantly one-sided investigations focusing on either of the partners involved. Here, we conducted a dual RNA-seq time course analysis of filarial worm parasite and host mosquito to better understand the parasite processes underlying development in and interaction with the host tissue, from the establishment of infection to the development of infective-stage larva.Using the Brugia malayi-Aedes aegypti system, we report parasite gene transcription dynamics, which exhibited a highly ordered developmental program consisting of a series of cyclical and state-transitioning temporal patterns. In addition, we contextualized these parasite data in relation to the concurrent dynamics of the host transcriptome. Comparative analyses using uninfected tissues and different host strains revealed the influence of parasite development on host gene transcription as well as the influence of the host environment on parasite gene transcription. We also critically evaluated the life-cycle transcriptome of B. malayi by comparing developmental stages in the mosquito relative to those in the mammalian host, providing insight into gene expression changes underpinning the mosquito-borne parasitic lifestyle of this heteroxenous parasite.The data presented herein provide the research community with information to design wet lab experiments and select candidates for future study to more fully dissect the whole set of molecular interactions of both organisms in this mosquito-filarial worm symbiotic relationship. Furthermore, characterization of the transcriptional program over the complete life cycle of the parasite, including stages within the mosquito, could help devise novel targets for control strategies

Crystal Structure and Substrate Specificity of Drosophila 3,4-Dihydroxyphenylalanine Decarboxylase

3,4-Dihydroxyphenylalanine decarboxylase (DDC), also known as aromatic L-amino acid decarboxylase, catalyzes the decarboxylation of a number of aromatic L-amino acids. Physiologically, DDC is responsible for the production of dopamine and serotonin through the decarboxylation of 3,4-dihydroxyphenylalanine and 5-hydroxytryptophan, respectively. In insects, both dopamine and serotonin serve as classical neurotransmitters, neuromodulators, or neurohormones, and dopamine is also involved in insect cuticle formation, eggshell hardening, and immune responses.In this study, we expressed a typical DDC enzyme from Drosophila melanogaster, critically analyzed its substrate specificity and biochemical properties, determined its crystal structure at 1.75 Angstrom resolution, and evaluated the roles residues T82 and H192 play in substrate binding and enzyme catalysis through site-directed mutagenesis of the enzyme. Our results establish that this DDC functions exclusively on the production of dopamine and serotonin, with no activity to tyrosine or tryptophan and catalyzes the formation of serotonin more efficiently than dopamine.The crystal structure of Drosophila DDC and the site-directed mutagenesis study of the enzyme demonstrate that T82 is involved in substrate binding and that H192 is used not only for substrate interaction, but for cofactor binding of drDDC as well. Through comparative analysis, the results also provide insight into the structure-function relationship of other insect DDC-like proteins

Detecting relic gravitational radiation from string cosmology with LIGO

A characteristic spectrum of relic gravitational radiation is produced by a period of ``stringy inflation" in the early universe. This spectrum is unusual, because the energy-density rises rapidly with frequency. We show that correlation experiments with the two gravitational wave detectors being built for the Laser Interferometric Gravitational Observatory (LIGO) could detect this relic radiation, for certain ranges of the parameters that characterize the underlying string cosmology model.Comment: 6 pages, 5 eps figures, Revte

Detection of Anisotropies in the Gravitational-Wave Stochastic Background

By correlating the signals from a pair of gravitational-wave detectors, one can undertake sensitive searches for a stochastic background of gravitational radiation. If the stochastic background is anisotropic, then this correlated signal varies harmonically with the earth's rotation. We calculate how the harmonics of this varying signal are related to the multipole moments which characterize the anisotropy, and give a formula for the signal-to-noise ratio of a given harmonic. The specific case of the two LIGO (Laser Interferometric Gravitational Observatory) detectors, which will begin operation around the year 2000, is analyzed in detail. We consider two possible examples of anisotropy. If the gravitational-wave stochastic background contains a dipole intensity anisotropy whose origin (like that of the Cosmic Background Radiation) is motion of our local system, then that anisotropy will be observable by the advanced LIGO detector (with 90% confidence in one year of observation) if \Omega_{gw} > 5.3 \times 10^{-8} h_{100}^{-2}. We also study the signal produced by stochastic sources distributed in the same way as the luminous matter in the galactic disk, and in the same way as the galactic halo. The anisotropy due to sources distributed as the galactic disk or as the galactic halo will be observable by the advanced LIGO detector (with 90% confidence in one year of observation) if \Omega_{gw} > 1.8 \times 10^{-10} h_{100}^{-2} or \Omega_{gw} > 6.7 \times 10^{-8} h_{100}^{-2}, respectively.Comment: 25 pages, Latex with RevTeX and epsfig, now includes S/N ratio calculations, expected response from anisotropy due to local motion & sources in galax

Persistence of Brugia malayi DNA in vector and non-vector mosquitoes: Implications for xenomonitoring and transmission monitoring of lymphatic filariasis

Abstract. Xenomonitoring (detection of filarial larvae or their DNA in mosquitoes) is a sensitive marker for as-sessing the endemicity of filariasis and a useful tool for evaluating elimination programs. To examine the fate of microfilariae (mf) and filarial DNA in vector competent and non-competent mosquito strains, we compared the detec-tion of Brugia malayi parasites by dissection and by quantitative real-time polymerase chain reaction (PCR) in three different mosquito strains. We conclude that PCR is much more sensitive than dissection for detecting filarial larvae, especially their remnants in mosquitoes. However, parasite DNA can be detected in both vector and non-vector mosquitoes for two weeks or longer after they ingest mf-positive blood. Thus, although xenomonitoring with vector and non-vector mosquito species may be a sensitive method for indirectly detecting filarial parasites in human populations, positive test results for parasite DNA in mosquitoes do not necessarily prove that transmission is ongoing in the study area

Mosquito transcriptome changes and filarial worm resistance in Armigeres subalbatus

<p>Abstract</p> <p>Background</p> <p><it>Armigeres subalbatus </it>is a natural vector of the filarial worm <it>Brugia pahangi</it>, but it rapidly and proficiently kills <it>Brugia malayi </it>microfilariae by melanotic encapsulation. Because <it>B. malayi </it>and <it>B. pahangi </it>are morphologically and biologically similar, the <it>Armigeres-Brugia </it>system serves as a valuable model for studying the resistance mechanisms in mosquito vectors. We have initiated transcriptome profiling studies in <it>Ar. subalbatus </it>to identify molecular components involved in <it>B. malayi </it>refractoriness.</p> <p>Results</p> <p>These initial studies assessed the transcriptional response of <it>Ar. subalbatus </it>to <it>B. malayi </it>at 1, 3, 6, 12, 24, 48, and 72 hrs after an infective blood feed. In this investigation, we initiated the first holistic study conducted on the anti-filarial worm immune response in order to effectively explore the functional roles of immune-response genes following a natural exposure to the parasite. Studies assessing the transcriptional response revealed the involvement of unknown and conserved unknowns, cytoskeletal and structural components, and stress and immune responsive factors. The data show that the anti-filarial worm immune response by <it>Ar. subalbatus </it>to be a highly complex, tissue-specific process involving varied effector responses working in concert with blood cell-mediated melanization.</p> <p>Conclusion</p> <p>This initial study provides a foundation and direction for future studies, which will more fully dissect the nature of the anti-filarial worm immune response in this mosquito-parasite system. The study also argues for continued studies with RNA generated from both hemocytes and whole bodies to fully expound the nature of the anti-filarial worm immune response.</p

A Deep Sequencing Approach to Comparatively Analyze the Transcriptome of Lifecycle Stages of the Filarial Worm, Brugia malayi

Lymphatic filariasis, also known as elephantiasis, is a tropical disease affecting over 120 million people worldwide. More than 40 million people live with painful, disfiguring symptoms that can cause severe debilitation and social stigma. The disease is caused by infection with thread-like filarial nematodes (roundworms) that have a complex parasitic lifecycle involving both human and mosquito hosts. In the study, the authors profiled the transcriptome (the set of genes transcribed into messenger RNA rather than all of those in the genome) of the human filarial worm Brugia malayi in different lifecyle stages using deep sequencing technology. The analysis revealed major transitions in RNA expression from eggs through larval stages to adults. Using statistical approaches, the authors identified groups of genes with distinct life stage dependent transcriptional patterns, with particular emphasis on genes displaying sex-biased or germline-enriched patterns and those displaying significant changes during larval development. This study presents a first comprehensive analysis of the lifecycle transcriptome of B. malayi, providing fundamental molecular information that should help researchers better understand parasite biology and could provide clues for the development of more effective interventions