Yellow fever virus envelope protein expressed in insect cells is capable of syncytium formation in lepidopteran cells and could be used for immunodetection of YFV in human sera

<p>Abstract</p> <p>Background</p> <p>Yellow fever is an haemorrhagic disease caused by a virus that belongs to the genus Flavivirus (Flaviviridae family) and is transmitted by mosquitoes. Among the viral proteins, the envelope protein (E) is the most studied one, due to its high antigenic potencial. Baculovirus are one of the most popular and efficient eukaryotic expression system. In this study a recombinant baculovirus (vSynYFE) containing the envelope gene (<it>env</it>) of the 17D vaccine strain of yellow fever virus was constructed and the recombinant protein antigenicity was tested.</p> <p>Results</p> <p>Insect cells infected with vSynYFE showed syncytium formation, which is a cytopathic effect characteristic of flavivirus infection and expressed a polypeptide of around 54 kDa, which corresponds to the expected size of the recombinant E protein. Furthermore, the recombinant E protein expression was also confirmed by fluorescence microscopy of vSynYFE-infected insect cells. Total vSynYFE-infected insect extracts used as antigens detected the presence of antibodies for yellow fever virus in human sera derived from yellow fever-infected patients in an immunoassay and did not cross react with sera from dengue virus-infected patients.</p> <p>Conclusions</p> <p>The E protein expressed by the recombinant baculovirus in insect cells is antigenically similar to the wild protein and it may be useful for different medical applications, from improved diagnosis of the disease to source of antigens for the development of a subunit vaccine.</p

Testing matter effects in propagation of atmospheric and long-baseline neutrinos

We quantify our current knowledge of the size and flavor structure of the matter effects in the evolution of atmospheric and long-baseline neutrinos based solely on the analysis of the corresponding neutrino data. To this aim we generalize the matter potential of the Standard Model by rescaling its strength, rotating it away from the e-e sector, and rephasing it with respect to the vacuum term. This phenomenological parametrization can be easily translated in terms of non-standard neutrino interactions in matter. We show that in the most general case, the strength of the potential cannot be determined solely by atmospheric and long-baseline data. However its flavor composition is very much constrained and the present determination of the neutrino masses and mixing is robust under its presence. We also present an update of the constraints arising from this analysis in the particular case in which no potential is present in the e-mu and e-tau sectors. Finally we quantify to what degree in this scenario it is possible to alleviate the tension between the oscillation results for neutrinos and antineutrinos in the MINOS experiment and show the relevance of the high energy part of the spectrum measured at MINOS.Comment: PDFLaTeX file using JHEP3 class, 25 pages, 7 figures included. Accepted for publication in JHE

Reactivity, performance, color and tenderness of meat from Zebu cattle finished in feedlot.

The objective of this study was to study the animal reactivity and correlate it with performance and meat quality of cattle finished in feedlots. A total of 20 animals of the Tabapuã breed and 20 animals of the Nellore breed were confined for 112 days: 28 days of adaptation and 84 days of data collection. Performance was determined by the average daily weight gain (ADG) and average dry matter intake (DMI) estimated through the use of indicators. On the 1st, 42nd and 84th days of the experiment, two methods were used to measure the animal reactivity: animal reactivity in mobile cage and visual reactivity during handling. The characteristics of meat quality were evaluated by the color, shear force and pH24 after slaughter (pH24). Through the study of correlation, the degree and direction of linear association on animal reactivity, performance and meat quality were established. Regression equations were generated for the parameters evaluated according to the values of animal reactivity. Correlations between mobile cage and visual reactivity were 0.77, 0.56 and 0.45 in the 1st, 2nd and 3rd evaluations, respectively. There was no correlation between animal reactivity and DMI. The correlations between ADG, pH24, and color with animal reactivity were -0.31, 0.40, 0.47 and -0.34, and -0.33 for mobile cage, 0.74, 0.71 and -0.51, for visual reactivity, respectively. The parameters of meat quality varied according to the reactivity. Undesirable values of pH24, animal reactivity and color were found when mobile cage reactivity was higher than 780, 590 and 540, respectively. In general, animals that show greater reactivity tend to have lower daily weight gain and lower meat quality. The reactivity can be used in animal breeding programs, in order to improve performance and meat quality of Zebu cattle finished in feedlot

Spectroscopic variability of two Oe stars

The Oe stars HD45314 and HD60848 have recently been found to exhibit very different X-ray properties: whilst HD60848 has an X-ray spectrum and emission level typical of most OB stars, HD45314 features a much harder and brighter X-ray emission, making it a so-called gamma Cas analogue. Monitoring the optical spectra could provide hints towards the origin of these very different behaviours. We analyse a large set of spectroscopic observations of HD45314 and HD60848, extending over 20 years. We further attempt to fit the H-alpha line profiles of both stars with a simple model of emission line formation in a Keplerian disk. Strong variations in the strengths of the H-alpha, H-beta, and He I 5876 emission lines are observed for both stars. In the case of HD60848, we find a time lag between the variations in the equivalent widths of these lines. The emission lines are double peaked with nearly identical strengths of the violet and red peaks. The H-alpha profile of this star can be successfully reproduced by our model of a disk seen under an inclination of 30 degrees. In the case of HD45314, the emission lines are highly asymmetric and display strong line profile variations. We find a major change in behaviour between the 2002 outburst and the one observed in 2013. This concerns both the relationship between the equivalent widths of the various lines and their morphologies at maximum strength (double-peaked in 2002 versus single-peaked in 2013). Our simple disk model fails to reproduce the observed H-alpha line profiles of HD45314. Our results further support the interpretation that Oe stars do have decretion disks similar to those of Be stars. Whilst the emission lines of HD60848 are explained by a disk with a Keplerian velocity field, the disk of HD45314 seems to have a significantly more complex velocity field that could be related to the phenomenon that produces its peculiar X-ray emission.Comment: Accepted for Publication in A&

The major leucyl aminopeptidase of Trypanosoma cruzi (LAPTc) assembles into a homohexamer and belongs to the M17 family of metallopeptidases

<p>Abstract</p> <p>Background</p> <p>Pathogens depend on peptidase activities to accomplish many physiological processes, including interaction with their hosts, highlighting parasitic peptidases as potential drug targets. In this study, a major leucyl aminopeptidolytic activity was identified in <it>Trypanosoma cruzi</it>, the aetiological agent of Chagas disease.</p> <p>Results</p> <p>The enzyme was isolated from epimastigote forms of the parasite by a two-step chromatographic procedure and associated with a single 330-kDa homohexameric protein as determined by sedimentation velocity and light scattering experiments. Peptide mass fingerprinting identified the enzyme as the predicted <it>T. cruzi </it>aminopeptidase EAN97960. Molecular and enzymatic analysis indicated that this leucyl aminopeptidase of <it>T. cruzi </it>(LAPTc) belongs to the peptidase family M17 or leucyl aminopeptidase family. LAPTc has a strong dependence on neutral pH, is mesophilic and retains its oligomeric form up to 80°C. Conversely, its recombinant form is thermophilic and requires alkaline pH.</p> <p>Conclusions</p> <p>LAPTc is a 330-kDa homohexameric metalloaminopeptidase expressed by all <it>T. cruzi </it>forms and mediates the major parasite leucyl aminopeptidolytic activity. Since biosynthetic pathways for essential amino acids, including leucine, are lacking in <it>T. cruzi</it>, LAPTc could have a function in nutritional supply.</p

The unconventional myosin CRINKLED and its mammalian orthologue MYO7A regulate caspases in their signalling roles

Caspases provide vital links in non-apoptotic regulatory networks controlling inflammation, compensatory proliferation, morphology and cell migration. How caspases are activated under non-apoptotic conditions and process a selective set of substrates without killing the cell remain enigmatic. Here we find that the Drosophila unconventional myosin CRINKLED (CK) selectively interacts with the initiator caspase DRONC and regulates some of its non-apoptotic functions. Loss of CK in the arista, border cells or proneural clusters of the wing imaginal discs affects DRONC-dependent patterning. Our data indicate that CK acts as substrate adaptor, recruiting SHAGGY46/GSK3-β to DRONC, thereby facilitating caspase-mediated cleavage and localized modulation of kinase activity. Similarly, the mammalian CK counterpart, MYO7A, binds to and impinges on CASPASE-8, revealing a new regulatory axis affecting receptor interacting protein kinase-1 (RIPK1)>CASPASE-8 signalling. Together, our results expose a conserved role for unconventional myosins in transducing caspase-dependent regulation of kinases, allowing them to take part in specific signalling events

Efeito da covariância genética aditivo-materna sobre estimativas de parâmetros genéticos e em avaliações genéticas de características de crescimento de animais Tabapuã.

RESUMO - Avaliou-se o efeito da covariância genética aditiva direto-materna sobre estimativas de parâmetros genéticos e sobre a predição e a ordenação de valores genéticos de animais da raça Tabapuã. Os parâmetros genéticos foram estimados com base em 19646, 14276, 10663 e 6172 registros de pesos ao nascimento e ajustados para 205, 365 e 550 dias de idade em análises unicaracterística, utilizando o programa computacional MTDREML, pelo método da máxima verossimilhança restrita livre de derivadas por modelos animal com ou sem a inclusão da covariância. As estimativas da covariância genética aditiva direto-materna foram, respectivamente, -0,08; -0,22; -0,10 e 0,34, para os pesos ao nascer e ajustados para 205, 365 e 550 dias de idade. As herdabilidades diretas e maternas obtidas sob modelo com a inclusão da covariância genética direto-materna foram, respectivamente, 0,31 e 0,10; 0,20 e 0,17; 0,20 e 0,06 e 0,17 e 0,01, para os mesmos pesos, enquanto sem a inclusão foram, respectivamente, 0,31 e 0,09; 0,18 e 0,14; 0,20 e 0,05 e 0,18 e 0,02. Os valores de correlação de posto entre os valores genéticos preditos pelos modelos com e sem a inclusão foram 0,999; 0,992; 0,999 e 0,998. As correlações de posto entre os valores genéticos maternos foram 0,999; 0,985; 0,992 e 0,771. A inclusão da covariância genética aditiva direto-materna não influenciou as estimativas dos parâmetros genéticos e teve efeito inexpressivo na ordenação dos valores genéticos dos pesos de animais da raça Tabapuã. ABSTRACT - This study was carried out to evaluate the effect of additive genetic-maternal covariance on estimates of genetic parameters and on the prediction and ranking of breeding values of Tabapuã animals. Records of 19,646, 14,276, 10,663, and 6,176 birth weights and weights at 205, 365, and 550 days of age were used in univariate animal model analysis using MTDFREML software, including or not the additive geneticmaternal covariance in the models. The additive genetic-maternal covariance estimates were, respectively, -0.08, -0.22, -0.10, and 0.34 for birth weight and weights at 205, 365, and 550 days of age. The direct and maternal heritability estimates including the additive direct-maternal covariance were, respectively, 0.31, and 0.10; 0.31 and 0.20, and 0.17; and 0.20 and 0.06; and 0.17 and 0.01 for birth weight and weights at 205, 365, and 550 days of age, while without the additive direct-maternal covariance those values were, respectively, 0.31 and 0.09; 0.18 and 0.14; 0.20 and 0.05; .18 and 0.02. Rank correlation between predicted breeding values from two models for birth weight and weights at 205 and 550 days of age were, respectively, 0.999, 0.992, 0.999, and 0.998. For maternal genetic values, these estimated rank correlations were, respectively, 0.995, 0.985, 0.992, and 0.771. The inclusion of additive genetic-maternal covariance in the analysis did not affect genetic parameter estimates and had a very small effect on breeding values ranking of Tabapuã animals

Non-standard interactions versus non-unitary lepton flavor mixing at a neutrino factory

The impact of heavy mediators on neutrino oscillations is typically described by non-standard four-fermion interactions (NSIs) or non-unitarity (NU). We focus on leptonic dimension-six effective operators which do not produce charged lepton flavor violation. These operators lead to particular correlations among neutrino production, propagation, and detection non-standard effects. We point out that these NSIs and NU phenomenologically lead, in fact, to very similar effects for a neutrino factory, for completely different fundamental reasons. We discuss how the parameters and probabilities are related in this case, and compare the sensitivities. We demonstrate that the NSIs and NU can, in principle, be distinguished for large enough effects at the example of non-standard effects in the $\mu$-$\tau$-sector, which basically corresponds to differentiating between scalars and fermions as heavy mediators as leading order effect. However, we find that a near detector at superbeams could provide very synergistic information, since the correlation between source and matter NSIs is broken for hadronic neutrino production, while NU is a fundamental effect present at any experiment.Comment: 32 pages, 5 figures. Final version published in JHEP. v3: Typo in Eq. (27) correcte

Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli

<p>Abstract</p> <p>Background</p> <p>Antigens for Hantavirus serological tests have been produced using DNA recombinant technology for more than twenty years. Several different strategies have been used for that purpose. All of them avoid the risks and difficulties involved in multiplying Hantavirus in the laboratory. In Brazil, the Araraquara virus is one of the main causes of Hantavirus Cardio-Pulmonary Syndrome (HCPS).</p> <p>Methods</p> <p>In this investigation, we report the expression of the N protein of the Araraquara Hantavirus in a Baculovirus Expression System, the use of this protein in IgM and IgG ELISA and comparison with the same antigen generated in <it>E. coli</it>.</p> <p>Results</p> <p>The protein obtained, and purified in a nickel column, was effectively recognized by antibodies from confirmed HCPS patients. Comparison of the baculovirus generated antigen with the N protein produced in <it>E. coli </it>showed that both were equally effective in terms of sensitivity and specificity.</p> <p>Conclusions</p> <p>Our results therefore indicate that either of these proteins can be used in serological tests in Brazil.</p