An Insight into the Sialome of the Lone Star Tick, \u3ci\u3eAmbylomma americanum\u3c/i\u3e, With a Glimpse On Its Time Dependent Gene Expression

Hard ticks feed for several days or weeks on their hosts. Blood feeding is assisted by tick saliva, which is injected in the host skin regularly, alternating with blood ingestion. Tick saliva contains hundreds or thousands of different peptides and other bioactive compounds that assist feeding by inhibiting their hosts’ blood clotting, platelet aggregation, vasoconstriction, as well as pain and itching. Immunomodulatory and antimicrobial peptides are also found in tick saliva. Molecular characterization of tick salivary compounds, or its sialome (from the Greek sialos = saliva), helps identification of possible antigens that might confer anti-tick immunity, as well as identifying novel pharmacologically active compounds. Amblyomma americanum is a major nuisance tick in Eastern and Southern US, being a vector of Theileria and Ehrlichia bacteria to animals and humans. Presently we report an RNAseq study concerning the salivary glands of adult female A. americanum ticks, which involved sequencing of four libraries collected at different times of feeding. A total of 5,792 coding sequences were deduced from the transcriptome assembly, 3,139 of which were publicly deposited, expanding from the previously available 146 salivary sequences found in GenBank. A remarkable time-dependent transcript expression was found, mostly related to secretory products, supporting the idea that ticks may have several “sialomes” that are expressed at different times during feeding. The molecular nature of this sialome switching remains unknown. The hyperlinked spreadsheet containing the deduced coding sequences can be found at http://exon.niaid.nih.gov/transcriptome/Amb_americanum/Ambame-web.xlsx

A Divergent Strain of Culex pipiens-Associated Tunisia Virus in the Malaria Vector Anopheles epiroticus

Here, we report the draft genome sequence of a divergent strain ofCulex pipiens-associated Tunisia virus (CpATV) identified in the malaria vectorAnoph-eles epiroticus(CpATV-AnE). CpATV-AnE expands the reference virus sequence, intro-ducing an extended replicase with novel virga-like domains. Our results suggest thatthe host range of CpATV includesAnophelessp. mosquitoes.Instituto de Patología VegetalFil: Debat, Humberto J. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal (IPAVE). Córdoba; ArgentinaFil: Ribero, Jose M C. National Institute of Allergy and Infectious Diseases (NIAID-NIH). Laboratory of Malaria and Vector Research.Section of Vector Biology; Estados Unido

A Deep Insight into the Sialotranscriptome of the Gulf Coast Tick, \u3ci\u3eAmblyomma maculatum\u3c/i\u3e

Background Saliva of blood sucking arthropods contains compounds that antagonize their hosts\u27 hemostasis, which include platelet aggregation, vasoconstriction and blood clotting; saliva of these organisms also has anti-inflammatory and immunomodullatory properties. Perhaps because hosts mount an active immune response against these compounds, the diversity of these compounds is large even among related blood sucking species. Because of these properties, saliva helps blood feeding as well as help the establishment of pathogens that can be transmitted during blood feeding. Methodology/Principal Findings We have obtained 1,626,969 reads by pyrosequencing a salivary gland cDNA library from adult females Amblyomma maculatum ticks at different times of feeding. Assembly of this data produced 72,441 sequences larger than 149 nucleotides from which 15,914 coding sequences were extracted. Of these, 5,353 had \u3e75% coverage to their best match in the non-redundant database from the National Center for Biotechnology information, allowing for the deposition of 4,850 sequences to GenBank. The annotated data sets are available as hyperlinked spreadsheets. Putative secreted proteins were classified in 133 families, most of which have no known function. Conclusions/Significance This data set of proteins constitutes a mining platform for novel pharmacologically active proteins and for uncovering vaccine targets against A. maculatum and the diseases they carry

Purification and characterization of a novel salivary antimicrobial peptide from the tick, \u3cem\u3eIxodes scapularis\u3c/em\u3e

A novel antimicrobial peptide was isolated from the saliva of the Lyme disease tick vector, Ixodes scapularis, henceforth designated as ISAMP (I. scapularis Antimicrobial Peptide). ISAMP was purified using a sequential method including ultra filtration, gel filtration and reverse-phase high performance liquid chromatography. The purified peak had a molecular weight of 5.3 kDa by MALDI/TOF-MS and its amino acid sequence, determined by Edman degradation was PDxGxPxxVKAGRxPxxSI. A BLASTP search revealed that the protein is a putative 5.3 kDa secreted protein (AAM93656) from I. scapularis. The predicted protein is composed of 69 amino acids with no conserved domain motifs. Purified ISAMP was found to have antimicrobial activities against bacteria. Gene expression studies were carried out to observe ISAMP expression in different tick tissues. RT-PCR results indicated that the gene was expressed in hemocytes, fat body and salivary gland but virtually no expression was observed in the midgut. ISAMP is only similar to other Ixodid tick proteins, thus it is a member of a unique family

Tick saliva is a potent inhibitor of endothelial cell proliferation and angiogenesis

We report for the first time that saliva of the hard tick and Lyme disease vector,Ixodes scapularis,is a potent inhibitor of angiogenesis. Saliva (≤ 1:500 dilutions) or salivary gland (0.1–0.5 pairs/assay) dose-dependently inhibits microvascular endothelial cell (MVEC) proliferation. Inhibition was also detected with the saliva of the cattle tick Boophilus microplus but not with the salivary gland of Anopheles gambiae, An. stephensi, Lutzomyia longipalpis, Phlebotomus papatasi, Aedes aegypti, Culex quinquefasciatus, and Cimex lectularius. Inhibition of MVEC proliferation by I. scapularis saliva was accompanied by a change in cell shape (shrinkage of the cytoplasm with loss of cell-cell interactions) and apoptosis which was estimated by expression of phosphatidylserine using the Apopercentage dye, and by a typical pattern of chromatin margination, condensation, and fragmentation as revealed by nuclear staining with Hoechst 33258.The effect of saliva appears to be mediated by endothelial cell α5β1 integrin, because monoclonal antibodies against this but not αvβ3, αvβ5, α9β1, or α2β1 integrins remarkably block its effect. In addition, SDS/PAGE shows that saliva specifically degrades purified α5β1 but not αvβ5 or αvβ3 integrins. Incubation of saliva with EDTA and 1,10-phenanthroline, but not phenylmethylsulfonyl fluoride (PMSF), inhibits saliva-dependent degradation of purified α5β1 integrin, suggesting that a metalloprotease is responsible for the activity. Finally, saliva at ≤ 1:1,000 dilutions blocks sprouting formation from chick embryo aorta implanted in Matrigel, an in vitro model of angiogenesis. These findings introduce the concept that tick saliva is a negative modulator of angiogenesis-dependent wound healing and tissue repair, therefore allowing ticks to feed for days. Inhibition of angiogenesis was hitherto an unidentified biologic property of the saliva of any blood-sucking arthropod studied so far. Its presence in tick saliva may be regarded as an additional source of angiogenesis inhibitors with potential applications for the study of both vector and vascular biology

Cloning of a salivary gland metalloprotease and characterization of gelatinase and fibrin(ogen)lytic activities in the saliva of the Lyme disease tick vector Ixodes scapularis

The full-length sequence of tick salivary gland cDNA coding for a protein similar to metalloproteases (MP) of the reprolysin family is reported. The Ixodes scapularis MP is a 488 amino acid (aa) protein containing pre- and pro-enzyme domains, the zinc-binding motif HExxHxxGxxH common to metalloproteases, and a cysteine-rich region. In addition, the predicted amino-terminal sequences of I. scapularis MPs were found by Edman degradation of PVDF-transferred SDS/PAGE-separated tick saliva proteins, indicating that these putative enzymes are secreted. Furthermore, saliva has a metal-dependent proteolytic activity towards gelatin, fibrin(ogen), and fibronectin, but not collagen or laminin. Accordingly, I. scapularis saliva has a rather specific metalloprotease similar to the hemorrhagic proteases of snake venoms. This is the first description of such activity in tick saliva and its role in tick feeding and Borrelia transmission is discussed

Changes in carbon stocks in Eucalyptus globulus Labill. plantations induced by different water and nutrient availability

Changes in the carbon stocks under different soil water and nutrient conditions were studied in Eucalyptus globulus Labill. stands in a field experiment, at O ´ bidos (central Portugal). The treatments were irrigation plus a complete fertiliser solution to simulate ‘near optimal’ nutrition (IF), irrigation only (I), and fertilisers added to rain-fed plots (F). The control (C) received neither water nor fertilisers (except a small amount at planting). The production of biomass (aboveground), the litterfall and the soil chemical composition were evaluated regularly during the experiment. Root biomass was estimated at the end of the experiment. Carbon in biomass, litterfall and soil, increased significantly when water and/or nutrients were supplied, in comparison to the control. The amount of carbon accumulated in the system, 6 years after planting, was 8.22, 10.22, 11.23 and 13.76 kg C m 2 in the control, F, I and IF treatments, respectively. The increase of carbon in the system during the same period was 5.86, 7.86, 8.87 and 11.40 kg C m 2 in the control, F, I and IF treatments, respectively. This rise in carbon resulted from the accumulation of long-lived woody biomass, which represented between 77.7 (in IF) and 82.9% (in the control) of the total rise in carbon. Although water was the main limiting factor for biomass accumulation, the allocation of carbon to the soil was mainly related to nutrient supply, irrespective of water availability. The amount of carbon stored belowground, i.e. soil and forest floor, plus stumps and roots, reached 4.2, 4.7, 4.8 and 6.3 kg C m 2 in the control, F, I and IF treatments, respectively. The increase in C in the mineral soil regarding the initial state was, in the same order as above, 0.21, 0.75, 0.58 and 1.21 kg C m 2. These values were 3.6, 9.6, 6.6 and 10.6% of the C accumulated in the whole system, during the experimental period

Deleção cromossómica intersticial em 14q “de novo”: apresentação de um caso

Introdução: As deleções intersticiais são anomalias cromossómicas estruturais, desequilibradas, resultantes de dois pontos de quebra, frequentemente associadas a quadros clínicos anormais devido à perda de material genético ativo (eucromatina). As consequências fenotípicas dependem do segmento cromossómico perdido e do número de genes aí localizados. Material e Métodos: Os autores apresentam o caso de um indivíduo do sexo masculino, de 11 anos de idade, referenciado para estudo citogenético por apresentar um quadro clínico de atraso de desenvolvimento psicomotor, défice cognitivo e problemas de comportamento. Realizaram-se culturas sincronizadas de linfócitos de sangue periférico, bandas GTG de alta resolução e, posteriormente, estudos de hibridação in situ por fluorescência (FISH) com sondas de pintura cromossómica total e subtelomérica, específicas para o cromossoma 14. Resultados: A análise das metafases revelou a presença de uma anomalia estrutural no cromossoma 14, interpretada como uma deleção intersticial do segmento compreendido entre as bandas 14q24.3 e 14q32.1. A análise por FISH permitiu confirmar esta deleção intersticial. Como os cariótipos dos pais foram normais, conclui-se que esta anomalia cromossómica é “de novo”, estabelecendo-se o cariótipo do doente como: 46,XY,del(14)(q24.3q32.1).ish del(14)(wcp 14+,SHGC36156+)dn Discussão: A deleção intersticial encontrada no cromossoma 14 implica uma monossomia do segmento 14q24.3→14q32.1. As alterações descritas mais comuns, associadas a esta deleção, incluem ADPM e algumas malformações minor. Os autores apresentam este caso pela raridade da anomalia citogenética encontrada e comparam-no com a literatura atual

Purification of a serine protease and evidence for a protein C activator from the saliva of the tick, \u3cem\u3eIxodes scapularis\u3c/em\u3e

The saliva of ticks is critical to their survival as parasites and hematophagous animals. In this study, we have purified an enzyme with trypsin-like activity from the saliva of the tick vector of Lyme Disease, Ixodes scapularis. This enzyme, named as IXOSP (I. scapularis salivary serine protease), is a 29.9 kDa molecule with N-terminus FPxMVxLRIKxR. A BLAST search identified IXOSP as a secreted serine protease (AAY66740) with a conserved catalytic triad His, Asp, and Ser. In vitro studies demonstrated that IXOSP cleaves chromogenic substrates with arginine in the P1 position, by a mechanism inhibited by PMSF or aprotinin. Gene expression studies revealed that IXOSP is expressed at different tick developmental stages, including eggs, and unfed or fed adult tick salivary glands, but not in nymphs or in the midgut. While the physiological substrate for IXOSP remains to be identified, we demonstrated that I. scapularis saliva activate protein C (PC) resulting in the production of activated PC, a potent anticoagulant that also regulates a myriad of inflammatory responses through protease activated receptors. In contrast, the salivary glands of Anopheles gambiae, Anopheles stephensi, Anopheles albimanus, Aedes aegypti, Lutzomyia longipalpis, and Phlebotomus ariasi did not activate protein C. These discoveries are discussed in the context of blood coagulation, inflammation and vector–host interactions

Ácido ascórbico, betalaínas, e fenóis totais em ecótipos de Opuntia spp.

O género Opuntia spp. pertence à família Cactaceae, sendo a espécie Opuntia ficus-indica (OFI) a que tem maior importância económica. Em frutos de vinte populações provenientes de quatro espécies do género Opuntia spp. (OFI, O. robusta, O. dillenii and O. elata), com origem em Portugal, foram estudadas as características cromáticas, a acidez, o pH, o teor em sólidos solúveis totais (SST) e ainda os teores em ácido ascórbico (AA), betalaínas e fenóis totais (FT). As cultivares Italianas de OFI ‘Bianca’, ‘Gialla’ e ‘Rossa’ foram incluídas como termo de comparação. Os valores mais elevados de acidez foram registados nos frutos de O. dillenii e O. elata e os menores em OFI. Os frutos de O. dillenii apresentaram os teores mais elevados de betalaínas e FT, ao passo que as concentrações mais elevadas de AA foram registadas nos frutos de O. elata. Os ecótipos de OFI mostraram variação na concentração de compostos bioativos. Em OFI, a cv. de polpa vermelha ‘Rossa’ é a que apresenta maior concentração de betalaínas, seguida pelos ecótipos de polpa laranja e, finalmente, os ecótipos de polpa branca. Os valores mais elevados de FT foram encontrados nos ecótipos de polpa branca. O género Opuntia spp. é uma fonte de compostos bioativos, pelo que o consumo dos seus frutos representa uma boa forma de ingestão de compostos de elevado valor nutricional.info:eu-repo/semantics/publishedVersio