Differential expression of Aedes aegypti salivary transcriptome upon blood feeding

Saliva of Aedes aegypti contains a complex array of proteins essential for both blood feeding and pathogen transmission. A large numbers of those proteins are classified as unknown in regard to their function(s). Understanding the dynamic interactions at the mosquito-host interface can be achieved in part by characterizing mosquito salivary gland gene expression relative to blood feeding. Towards this end, we developed an oligonucleotide microarray representing 463 transcripts to determine differential regulation of salivary gland genes. This microarray was used to investigate the temporal gene expression pattern of Ae. aegypti salivary gland transcriptome at different times post-blood feeding. Expression of the majority of salivary gland genes (77–87%) did not change significantly as a result of blood feeding, while 8 to 20% of genes were down-regulated and 2.8 to 11.6% genes were up-regulated. Up-regulated genes included defensins, mucins and other immune related proteins. Odorant-binding protein was significantly down-regulated. Among unknown function proteins, several were up-regulated during the first three hours post-blood feeding and one was significantly down-regulated. Quantitative real-time RT-PCR was used to substantiate differential expression patterns of five randomly selected genes. Linear regression analysis revealed a high degree of correlation (R2 > 0.89) between oligonucleotide microarray and quantitative RT-PCR data. To our knowledge, this is the first study to investigate differential expression of the Ae. aegypti salivary gland transcriptome upon blood feeding. A microarray provides a robust, sensitive way to investigate differential regulation of mosquito salivary gland genes

Chronic Health Conditions, School Attendance, and Socioeconomic Factors: A Literature Review

Students with chronic health conditions experience many challenges, such as increased school absences, that may impact academic success. Additional academic problems exist for students with chronic health conditions who also have socioeconomic status (SES) factors. A systematic literature review as conducted to study the interactions between chronic health conditions, absenteeism, and factors associated with SES. Findings suggest that asthma, the most common chronic health condition in children, is the leading cause of health-related school absences. Findings also suggest that factors associated with SES, such as family income, also impact school attendance for students with chronic health conditions. Although the results are mixed for academic achievement abilities for student who have chronic health conditions, many academic challenges that students with chronic health conditions encounter can be explained by frequent school absences. Further research on school absences and SES factors affecting students with chronic illnesses is needed to help establish guidelines for appropriate school programming and positive school attendance support

Purification, Cloning, and Expression of a Novel Salivary Anticomplement Protein from the Tick, Ixodes scapularis

The alternative pathway of complement is an important defense against pathogens and in tick rejection reactions. The tick Ixodes scapularis is able to feed repeatedly on its natural host and has a salivary anticomplement activity that presumably facilitates feeding. In this study, we purified and then obtained the amino-terminal sequence of the I. scapularissalivary anticomplement (Isac). We found a full-length clone coding for Isac by random screening of a salivary gland cDNA library. Expressing Isac cDNA in COS cells reproduced the activity found in tick saliva, namely, inhibition of rabbit erythrocyte lysis by human serum in the presence of Mg2+ and EGTA, inhibition of C3b binding to agarose in the presence of Mg2+ and EGTA, and acceleration of factor Bb uncoupling from the C3 convertase generated by the alternative pathway. Recombinant Isac had no effect on the recalcification time of human platelet-poor plasma or in the classical complement pathway, indicating that it is a specific inhibitor similar to the regulators of complement activation of the alternative pathway such as factor H. Isac, however, has no similarity to any protein in the GenBankTM data base, indicating that it is a novel and relatively small (18.5 kDa) anticomplement molecule

PENGARUH PENGGUNAAN SIARAN RADIO MALAYSIA TERHADAP MINAT MASYARAKAT DESA SELATBARU BENGKALIS DALAM MENDENGARKAN RADIO LOKAL

ABSTRAK Nama : T. Wikel Novryan Nim : 11840313940 Judul : Pengaruh Penggunaan Siaran Radio Malaysia Terhadap Minat Masyarakat Desa Selatbaru Bengkalis Dalam Mendengarkan Radio Lokal Penelitian ini dilatarbelakangi oleh kesenjangan informasi yang terjadi pada masyarakat yang tinggal diwilayah perbatasan desa selatbaru bengkalis. Dengan kurangnya insfrastuktur penyiaran di perbatasan, membuat masyarakat diwilayah desa selatbaru menjadikan siaran radio Malaysia dan radio lokal sebagai sebuah opsi pilihan untuk mendapatkan informasi yang dibutuhkan. Pendekatan yang digunakan dalam penelitian ini adalah kuantitatif. Tujuan dari penelitian ini adalah untuk mengetahui seberapa besar pengaruh penggunaan siaran radio Malaysia terhadap minat masyrakat desa selatbaru bengkalis mendengarakan radio lokal. Populasi pada penelitian ini adalah masyarakat desa selatbaru bengkalis berjumlah 3.940 jiwa. Teknik pengambilan sampel yang digunakan adalah metode purposive sampling, berdasarkan kriteria tertenti menggunakan rumus slovin, didapatkan sampel sebanyak 100 orang. Penelitian ini menggunakan Teknik metode angket (kuisioner) sebagai instrument utama dalam pengumpulan data. Teori yang digunakan pada penelitian ini adalah Uses and Gratifications. Hasil dari penelitian berdasarakan hasil analisis, perhitungan koefisien korelasi dan uji signifikan pengaruh variabel x (penggunaan sradio Malaysia) terhadap variabel Y (Minat Masyarakat), diketahui bahwa koefisien korelasi 0,796 yang menunjukkan data bahwa berpengaruh kuat terhadap minat masyarakat, Hasil dari perhitungan besaran pengaruh penggunaan siaran radio Malaysia terhadap minat masyarakat desa selatbaru bengkalis dalam mendengarkan radio lokal, berada pada 79,6%Maka dari itu dapat disimpulkan bahwa dengan masyarakat sering menggunakan siaran radio Malaysia dalam keseharian dapat mempengaruhi minat masyarakat dalam mendengarkan radio lokal disebabkan oleh beberapa faktor yang mempengaruhi. Kata Kunci : Radio, Tingkat Penggunaan Media, Minat Mendengarakan Radio

Atlantic Coast and Inner Shelf

The continental margin of Virginia, and of North America more broadly, is the physical transition from the high elevation of the continent to the low of the ocean basin. This transition was created as rifting pulled apart the ancient supercontinent Pangaea to create the Atlantic Ocean basin. Tectonic forces fractured and stretched the bedrock to create a stair-step ramp that subsequently would be mantled with sediment built up by erosion and transport off the continent. The Coastal Plain and Continental Shelf of Virginia are contiguous and discrete physiographic provinces of the continental margin delimited by the present elevation of sea level. On geologic time scales of thousands to millions of years, the coastal zone—the boundary between the coastal plain and shelf—is dynamic and migrates hundreds of kilometers landward and seaward. Today, the Atlantic shore of Virginia lies just past halfway across the margin: about 150 km (93 mi) from the edge of the Piedmont at the Fall Zone, and about 100 km (62 mi) from the seaward edge of the shelf (Figure 1). The modern coastal zone occupies nearly the same position as during several previous interglacial highstands of sea level that have recurred at approximately 100,000-year (abbreviated 100 ky, for “kilo year”) intervals since the middle Pleistocene (about the last 750 ky). more ...https://scholarworks.wm.edu/vimsbooks/1116/thumbnail.jp

Transcriptional profiling of the murine cutaneous response during initial and subsequent infestations with Ixodes scapularis nymphs

<p>Abstract</p> <p>Background</p> <p><it>Ixodes scapularis </it>ticks are hematophagous arthropods capable of transmitting many infectious agents to humans. The process of blood feeding is an extended and continuous interplay between tick and host responses. While this process has been studied extensively <it>in vitro</it>, no global understanding of the host response to ticks has emerged.</p> <p>Methods</p> <p>To address this issue, we used PCR-arrays to measure skin-specific expression of 233 discrete genes at 8 time points during primary and secondary infestations of mice with pathogen-free <it>I. scapularis </it>nymphs. Selected results were then validated at the mRNA and protein levels by additional real-time PCR and bioplex assay.</p> <p>Results</p> <p>Primary infestation was characterized by the late induction of an innate immune response. Lectin pattern recognition receptors, cytokines, and chemokines were upregulated consistent with increased neutrophil and macrophage migration. Gene ontology and pathway analyses of downregulated genes suggested inhibition of gene transcription and Th17 immunity. During the secondary infestation, additional genes were modulated suggesting a broader involvement of immune cells including CD8 and CD4 positive T lymphocytes. The cytokine response showed a mixed Th1/Th2 profile with a potential for T regulatory cell activity. Key gene ontology clusters observed during the secondary infestation were cell migration and activation. Matrix metalloproteinases were upregulated, apoptosis-related genes were differentially modulated, and immunoreceptor signaling molecules were upregulated. In contrast, transcripts related to mitogenic, WNT, Hedgehog, and stress pathways were downregulated.</p> <p>Conclusions</p> <p>Our results support a model of tick feeding where lectin pattern recognition receptors orchestrate an innate inflammatory response during primary infestation that primes a mixed Th1/Th2 response upon secondary exposure. Tick feeding inhibits gene transcription and Th17 immunity. Salivary molecules may also inhibit upregulation of mitogenic, WNT, Hedgehog, and stress pathways and enhance the activity of T regulatory cells, production of IL-10, and suppressors of cytokine signaling molecules (SOCS). This study provides the first comprehensive transcriptional analysis of the murine host response at the <it>I. scapularis </it>bite site and suggests both a potential model of the host cutaneous response and candidate genes for further description and investigation.</p

Changes in Temporal and Spatial Patterns of Outer Surface Lipoprotein Expression Generate Population Heterogeneity and Antigenic Diversity in the Lyme Disease Spirochete, Borrelia burgdorferi

This is the published version. Copyright 2015 by the American Society for Microbiology.Borrelia burgdorferi differentially expresses many of the OspE/F/Elp paralogs during tick feeding. These findings, combined with the recent report that stable B. burgdorferi infection of mammals occurs only after 53 h of tick attachment, prompted us to further analyze the expression of the OspE/F/Elp paralogs during this critical period of transmission. Indirect immunofluorescence analysis revealed that OspE, p21, ElpB1, ElpB2, and OspF/BbK2.11 are expressed in the salivary glands of ticks allowed to feed on mice for 53 to 58 h. Interestingly, many of the spirochetes in the salivary glands that expressed abundant amounts of these antigens were negative for OspA and OspC. Although prior reports have indicated that OspE/F/Elp orthologs are surface exposed, none of the individual lipoproteins or combinations of the lipoproteins protected mice from challenge infections. To examine why these apparently surface-exposed lipoproteins were not protective, we analyzed their genetic stability during infection and their cellular locations after cultivation in vitro and within dialysis membrane chambers, mimicking a mammalian host-adapted state. Combined restriction fragment length polymorphism and nucleotide sequence analyses revealed that the genes encoding these lipoproteins are stable for at least 8 months postinfection. Interestingly, cellular localization experiments revealed that while all of these proteins can be surface localized, there were significant populations of spirochetes that expressed these lipoproteins only in the periplasm. Furthermore, host-specific signals were found to alter the expression patterns and final cellular location of these lipoproteins. The combined data revealed a remarkable heterogeneity in populations of B. burgdorferi during tick transmission and mammalian infection. The diversity is generated not only by temporal changes in antigen expression but also by modulation of the surface lipoproteins during infection. The ability to regulate the temporal and spatial expression patterns of lipoproteins throughout infection likely contributes to persistent infection of mammals by B. burgdorferi

Rhipicephalus microplus salivary gland molecules induce differential CD86 expression in murine macrophages

<p>Abstract</p> <p>Background</p> <p>Tick parasitism is a major impediment for cattle production in many parts of the world. The southern cattle tick, <it>Rhipicephalus </it>(<it>Boophilus</it>) <it>microplus</it>, is an obligate hematophagous parasite of domestic and wild animals that serves as vector of infectious agents lethal to cattle. Tick saliva contains molecules evolved to modulate host innate and adaptive immune responses which facilitates blood feeding and pathogen transmission. Tick feeding promotes CD4 T cell polarization to a Th2 profile usually accompanied by down-regulation of Th1 cytokines through as yet undefined mechanisms. Co-stimulatory molecules on antigen presenting cells are central to development of T cell responses including Th1 and Th2 responses. Tick induced changes to antigen presenting cell signal transduction pathways are largely unknown. Here we document the ability of <it>R</it>. <it>microplus </it>salivary gland extracts (SGE) to effect differential CD86 expression.</p> <p>Results</p> <p>We examined changes in co-stimulatory molecule expression in murine RAW 264.7 cells in response to <it>R</it>. <it>microplus </it>SGE exposure in the presence of the toll-like receptor 4 (TLR4) ligand, LPS. After 24 hrs, CD86, but not CD80, was preferentially up-regulated on mouse macrophage RAW 264.7 cells when treated with SGE and then LPS, but not SGE alone. CD80 and CD40 expression was increased with LPS, but the addition of SGE did not alter expression. Higher concentrations of SGE were less effective at increasing CD86 RNA expression. The addition of mitogen or extracellular kinase (MEK) inhibitor, PD98059, significantly reduced the ability for SGE to induce CD86 expression, indicating activation of MEK is necessary for SGE induced up-regulation.</p> <p>Conclusions</p> <p>Molecules in SGE of <it>R. microplus </it>have a concentration-dependent effect on differential up-regulation of CD86 in a macrophage cell line activated by the TLR4 ligand, LPS. This CD86 up-regulation is at least partially dependent on the ERK1/2 pathway and may serve to promote Th2 polarization of the immune response.</p

An annotated catalogue of salivary gland transcripts in the adult female mosquito, Ædes ægypti*

BACKGROUND: Saliva of blood-sucking arthropods contains a cocktail of antihemostatic agents and immunomodulators that help blood feeding. Mosquitoes additionally feed on sugar meals and have specialized regions of their glands containing glycosidases and antimicrobials that might help control bacterial growth in the ingested meals. To expand our knowledge on the salivary cocktail of Ædes ægypti, a vector of dengue and yellow fevers, we analyzed a set of 4,232 expressed sequence tags from cDNA libraries of adult female mosquitoes. RESULTS: A nonredundant catalogue of 614 transcripts (573 of which are novel) is described, including 136 coding for proteins of a putative secretory nature. Additionally, a two-dimensional gel electrophoresis of salivary gland (SG) homogenates followed by tryptic digestion of selected protein bands and MS/MS analysis revealed the expression of 24 proteins. Analysis of tissue-specific transcription of a subset of these genes revealed at least 31 genes whose expression is specific or enriched in female SG, whereas 24 additional genes were expressed in female SG and in males but not in other female tissues. Most of the 55 proteins coded by these SG transcripts have no known function and represent high-priority candidates for expression and functional analysis as antihemostatic or antimicrobial agents. An unexpected finding is the occurrence of four protein families specific to SG that were probably a product of horizontal transfer from prokaryotic organisms to mosquitoes. CONCLUSION: Overall, this paper contributes to the novel identification of 573 new transcripts, or near 3% of the Æ. ægypti proteome assuming a 20,000-protein set, and to the best-described sialome of any blood-feeding insect