An RNA interference approach for functional studies in the sea urchin and its use in analysis of Nodal signaling gradients.

Dicer substrate interfering RNAs (DsiRNAs) destroy targeted transcripts using the RNA-Induced Silencing Complex (RISC) through a process called RNA interference (RNAi). This process is ubiquitous among eukaryotes. Here we report the utility of DsiRNA in embryos of the sea urchin Lytechinus variagatus (Lv). Specific knockdowns phenocopy known morpholino and inhibitor knockdowns, and DsiRNA offers a useful alternative to morpholinos. Methods for designing and obtaining specific DsiRNAs that lead to destruction of targeted mRNA are described. DsiRNAs directed against pks1 , an enzyme necessary for pigment production, show how successful DsiRNA perturbations are monitored by RNA in situ analysis and by qPCR to determine relative destruction of targeted mRNA. DsiRNA-based knockdowns phenocopy morpholino- and drug-based inhibition of nodal and lefty . Other knockdowns demonstrate that the RISC operates early in development as well as on genes that are first transcribed hours after gastrulation is completed. Thus, DsiRNAs effectively mediate destruction of targeted mRNA in the sea urchin embryo. The approach offers significant advantages over other widely used methods in the urchin in terms of cost, and ease of procurement, and offers sizeable experimental advantages in terms of ease of handling, injection, and knockdown validation. Highlights: DsiRNA provides an RNAi approach for perturbation of sea urchin embryos. A dilution series of DsiRNA oligos reveals properties of the Nodal gradient in establishing the Dorsal-Ventral axis.

Análisis in silico de las alteraciones de la secuencia y estructura de RBD-SARS-CoV-2, que afectan su complementariedad por anticuerpos neutralizantes IgG-antiRBD

Las vacunas anti-SARS-CoV-2 inducen la producción de anticuerpos neutralizantes IgG contra el Dominio de Unión al Receptor de la proteína S del virus (IgG-antiRBD). En Bolivia, Sinopharm y Sputnik V fueron vacunas ampliamente utilizadas durante la pandemia. Sin embargo, las mutaciones y los cambios sufridos en SARS-CoV-2 fueron responsables de las nuevas olas de contagio. Objetivo: determinar las alteraciones a nivel de secuencia y de estructura del RBD-SARS-CoV-2, que afectan su complementariedad por anticuerpos neutralizantes IgG-antiRBD. Material y Métodos: se obtuvieron las secuencias y estructuras cristalográficas del RBDSARS-CoV-2 a partir de la base de datos Protein Data Bank. Para el Alineamiento Múltiple de Secuencias y el Alineamiento Estructural, se emplearon Mega6 y Chimera1,15. Resultados: el Alineamiento Múltiple de Secuencias y Alineamiento Estructural de las principales variantes epidemiológicas de SARS-CoV-2 evidencian que, krakenXBB1.5 fue la más divergente a nivel de secuencia, mientras que, omicronBA2.75 presentó más cambios estructurales y mayores impedimentos estéricos al interaccionar con IgG-antiRBD, siendo la más contagiosas y más evasiva a la respuesta inmunológica. Conclusiones: el uso de herramientas bioinformáticas para el seguimiento en los cambios moleculares de SARS-CoV-2 permiten predecir el comportamiento epidemiológico de nuevas variantes emergentes y además promover el mejoramiento en los criterios de prevención.

Anti-SARS-CoV-2 vaccines induce the production of IgG neutralizing antibodies against the Receptor Binding Domain of the S protein of the virus (IgG-antiRBD). In Bolivia, Sinopharm and Sputnik V were widely used vaccines during the pandemic. However, the mutations and changes suffered in SARS-CoV-2 were responsible for the new contagion outbreaks. Objective: To determine the sequence and structure alterations of RBD-SARS-CoV-2, which affect its complementarity by IgG-antiRBD neutralizing antibodies. Material and Methods: The sequences and crystallographic structures of RBD-SARS-CoV-2 were obtained from the Protein Data Bank database. For Multiple Sequence Alignment and Structural Alignment, Mega6 and Chimera1.15 were used. Results: The Multiple Sequence Alignment and Structural Alignment of the main epidemiological variants of SARS-CoV-2 showed that krakenXBB1.5 was the most divergent at the sequence level, while omicronBA2.75 presented more structural changes and greater steric hindrances when interacting with IgG-antiRBD, being the most contagious and most evasive to the immune response. Conclusions: The use of bioinformatics tools for monitoring the molecular changes of SARS-CoV-2 make it possible to predict the epidemiological behavior of new emerging variants and also promote improvement in prevention criteria

Wound repair in sea urchin larvae involves pigment cells and blastocoelar cells.

Sea urchin larvae spend weeks to months feeding on plankton prior to metamorphosis. When handled in the laboratory they are easily injured, suggesting that in the plankton they are injured with some frequency. Fortunately, larval wounds are repaired through an efficient wound response with mesenchymal pigment cells and blastocoelar cells assisting as the epithelium closes. An injury to the epithelium leads to an immediate calcium transient that rapidly spreads around the entire larva and is necessary for activating pigment cell migration toward the wound. If calcium transport is blocked, the pigment cells fail to activate and remain in place. When activated, pigment cells initiate directed migration to the wound site from distances of at least 85 â€‹µm. Upon arrival at the wound site they participate in an innate immune response. Blastocoelar cells are recruited to the injury site as well, though the calcium transient is unnecessary for activating these cells. At the wound site, blastocoelar cells participate in several functions including remodeling the skeleton if it protrudes through the epithelium.

High burden of soil-transmitted helminth infections, schistosomiasis, undernutrition, and poor sanitation in two Typhoon Haiyan-stricken provinces in Eastern Philippines.

This study aimed to describe: 1) soil-transmitted helminthiasis (STH) and schistosomiasis prevalence and intensity in preschool-age children (PSAC) and school-age children (SAC), 2) schistosomiasis seroprevalence in SAC, 3) undernutrition prevalence in SAC, 4) sanitary toilet coverage in households, and 5) association between STH, Schistosoma japonicum exposure, nutritional status, and sanitation. PSAC and SAC in two Haiyan-stricken provinces were examined using Kato-Katz technique and ELISA Antibody test. Anthropometric and hemoglobin measurements were also obtained. The reported sanitary toilet coverage was validated in a survey. The prevalence of any STH in PSAC and SAC was 50.2% and 41.3%, respectively. Moderate-heavy intensity (MHI) STH prevalence in PSAC and SAC was 20.8% and 5.9%, respectively. The prevalence of any STH, MHI STH, ascariasis, MHI ascariasis, and MHI trichuriasis was significantly higher in PSAC. Stunting, underweight, wasting, overweight/obesity, and anemia prevalence was 38.4%, 24.5%, 4.8%, 2.7%, and 34.7% in PSAC, while the prevalence was 34.3%, 21.6%, 8.7%, 3.0%, and 19.2% in SAC, respectively. Anemia and wasting prevalence were significantly higher in PSAC and SAC, respectively. There were five schistosomiasis cases found (0.8% prevalence), while schistosomiasis seroprevalence was 60.1%. Validated and reported sanitary toilet coverage was significantly different in eight out of 13 barangays. Stunting and anemia were associated with STH. STH and anemia prevalence were significantly higher in non-ZOD barangays. High STH burden in PSAC and SAC persists. A more coordinated response addressing STH, undernutrition, and WASH in disaster-stricken areas will require strengthening local health systems and promoting intersectoral collaboration.

Gastrulation in the sea urchin.

Gastrulation is arguably the most important evolutionary innovation in the animal kingdom. This process provides the basic embryonic architecture, an inner layer separated from an outer layer, from which all animal forms arise. An extraordinarily simple and elegant process of gastrulation is observed in the sea urchin embryo. The cells participating in sea urchin gastrulation are specified early during cleavage. One outcome of that specification is the expression of transcription factors that control each of the many subsequent morphogenetic changes. The first of these movements is an epithelial-mesenchymal transition (EMT) of skeletogenic mesenchyme cells, then EMT of pigment cell progenitors. Shortly thereafter, invagination of the archenteron occurs. At the end of archenteron extension, a second wave of EMT occurs to release immune cells into the blastocoel and primordial germ cells that will home to the coelomic pouches. The archenteron then remodels to establish the three parts of the gut, and at the anterior end, the gut fuses with the stomodaeum to form the through-gut. As part of the anterior remodeling, mesodermal coelomic pouches bud off the lateral sides of the archenteron tip. Multiple cell biological processes conduct each of these movements and in some cases the upstream transcription factors controlling this process have been identified. Remarkably, each event seamlessly occurs at the right time to orchestrate formation of the primitive body plan. This review covers progress toward understanding many of the molecular mechanisms underlying this sequence of morphogenetic events.

Developmental origin of peripheral ciliary band neurons in the sea urchin embryo.

In the sea urchin larva, most neurons lie within an ectodermal region called the ciliary band. Our understanding of the mechanisms of specification and patterning of these peripheral ciliary band neurons is incomplete. Here, we first examine the gene regulatory landscape from which this population of neural progenitors arise in the neuroectoderm. We show that ciliary band neural progenitors first appear in a bilaterally symmetric pattern on the lateral edges of chordin expression in the neuroectoderm. Later in development, these progenitors appear in a salt-and-pepper pattern in the ciliary band where they express soxC, and prox, which are markers of neural specification, and begin to express synaptotagminB, a marker of differentiated neurons. We show that the ciliary band expresses the acid sensing ion channel gene asicl, which suggests that ciliary band neurons control the larva's ability to discern touch sensitivity. Using a chemical inhibitor of MAPK signaling, we show that this signaling pathway is required for proper specification and patterning of ciliary band neurons. Using live imaging, we show that these neural progenitors undergo small distance migrations in the embryo. We then show that the normal swimming behavior of the larvae is compromised if the neurogenesis pathway is perturbed. The developmental sequence of ciliary band neurons is very similar to that of neural crest-derived sensory neurons in vertebrates and may provide insights into the evolution of sensory neurons in deuterostomes.

Whole mount in situ hybridization techniques for analysis of the spatial distribution of mRNAs in sea urchin embryos and early larvae.

A critical process in embryonic development is the activation and spatial localization of mRNAs to specific cells and territories of the embryo. Revealing the spatial distribution of mRNAs and how it changes during development is a vital piece of information that aids in understanding the signaling and regulatory genes driving specific gene regulatory networks. In the laboratory, a cost-efficient, reliable method to determine the spatial distribution of mRNAs in embryos is in situ hybridization. This sensitive and straightforward method employs exogenous antisense RNA probes to find specific and complementary sequences in fixed embryos. Antigenic moieties conjugated to the ribonucleotides incorporated in the probe cross-react with antibodies, and numerous staining methods can be subsequently employed to reveal the spatial distribution of the targeted mRNA. The quality of the data produced by this method is equivalent to the experience of the researcher, and thus a thorough understanding of the numerous steps comprising this method is important for obtaining high quality data. Here we compile and summarize several protocols that have been employed chiefly on five sea urchin species in numerous laboratories around the world. Whereas the protocols can vary for the different species, the overarching steps are similar and can be readily mastered. When properly and carefully undertaken, in situ hybridization is a powerful tool providing unambiguous data for which there currently is no comparable substitute and will continue to be an important method in the era of big data and beyond.

Spatial and temporal patterns of gene expression during neurogenesis in the sea urchin Lytechinus variegatus.

BACKGROUND: The sea urchin is a basal deuterostome that is more closely related to vertebrates than many organisms traditionally used to study neurogenesis. This phylogenetic position means that the sea urchin can provide insights into the evolution of the nervous system by helping resolve which developmental processes are deuterostome innovations, which are innovations in other clades, and which are ancestral. However, the nervous system of echinoderms is one of the least understood of all major metazoan phyla. To gain insights into echinoderm neurogenesis, spatial and temporal gene expression data are essential. Then, functional data will enable the building of a detailed gene regulatory network for neurogenesis in the sea urchin that can be compared across metazoans to resolve questions about how nervous systems evolved. RESULTS: Here, we analyze spatiotemporal gene expression during sea urchin neurogenesis for genes that have been shown to be neurogenic in one or more species. We report the expression of 21 genes expressed in areas of neurogenesis in the sea urchin embryo from blastula stage (just before neural progenitors begin their specification sequence) through pluteus larval stage (when much of the nervous system has been patterned). Among those 21 gene expression patterns, we report expression of 11 transcription factors and 2 axon guidance genes, each expressed in discrete domains in the neuroectoderm or in the endoderm. Most of these genes are expressed in and around the ciliary band. Some including the transcription factors Lv-mbx, Lv-dmrt, Lv-islet, and Lv-atbf1, the nuclear protein Lv-prohibitin, and the guidance molecule Lv-semaa are expressed in the endoderm where they are presumably involved in neurogenesis in the gut. CONCLUSIONS: This study builds a foundation to study how neurons are specified and evolved by analyzing spatial and temporal gene expression during neurogenesis in a basal deuterostome. With these expression patterns, we will be able to understand what genes are required for neural development in the sea urchin. These data can be used as a starting point to (1) build a spatial gene regulatory network for sea urchin neurogenesis, (2) identify how subtypes of neurons are specified, (3) perform comparative studies with the sea urchin, protostome, and vertebrate organisms.

Neurogenesis in the sea urchin embryo is initiated uniquely in three domains.

Many marine larvae begin feeding within a day of fertilization, thus requiring rapid development of a nervous system to coordinate feeding activities. Here, we examine the patterning and specification of early neurogenesis in sea urchin embryos. Lineage analysis indicates that neurons arise locally in three regions of the embryo. Perturbation analyses showed that when patterning is disrupted, neurogenesis in the three regions is differentially affected, indicating distinct patterning requirements for each neural domain. Six transcription factors that function during proneural specification were identified and studied in detail. Perturbations of these proneural transcription factors showed that specification occurs differently in each neural domain prior to the Delta-Notch restriction signal. Though gene regulatory network state changes beyond the proneural restriction are largely unresolved, the data here show that the three neural regions already differ from each other significantly early in specification. Future studies that define the larval nervous system in the sea urchin must therefore separately characterize the three populations of neurons that enable the larva to feed, to navigate, and to move food particles through the gut.

Contribution of hedgehog signaling to the establishment of left-right asymmetry in the sea urchin.

Most bilaterians exhibit a left-right asymmetric distribution of their internal organs. The sea urchin larva is notable in this regard since most adult structures are generated from left sided embryonic structures. The gene regulatory network governing this larval asymmetry is still a work in progress but involves several conserved signaling pathways including Nodal, and BMP. Here we provide a comprehensive analysis of Hedgehog signaling and it's contribution to left-right asymmetry. We report that Hh signaling plays a conserved role to regulate late asymmetric expression of Nodal and that this regulation occurs after Nodal breaks left-right symmetry in the mesoderm. Thus, while Hh functions to maintain late Nodal expression, the molecular asymmetry of the future coelomic pouches is locked in. Furthermore we report that cilia play a role only insofar as to transduce Hh signaling and do not have an independent effect on the asymmetry of the mesoderm. From this, we are able to construct a more complete regulatory network governing the establishment of left-right asymmetry in the sea urchin.

Association of killer cell immunoglobulin-like receptor polymorphisms with chronic hepatitis C and responses to therapy in Brazil.

Soroprevalence for Hepatitis C virus is reported as 2.12% in Northern Brazil, with about 50% of the patients exhibiting a sustained virological response (SVR). Aiming to associate polymorphisms in Killer Cell Immunoglobulin-like Receptors (KIR) with chronic hepatitis C and therapy responses we investigated 125 chronic patients and 345 controls. Additionally, 48 ancestry markers were genotyped to control for population stratification. The frequency of the KIR2DL2 and KIR2DL2+HLA-C(Asp80) gene and ligand was higher in chronic infected patients than in controls (p < 0.0009, OR = 3.4; p = 0.001, OR = 3.45). In fact, KIR2DL3 is a weaker inhibitor of NK activity than KIR2DL2, which could explain the association of KIR2DL2 with chronic infection. Moreover, KIR2DS2 and KIR2DS2+HLA-C(Asp80) (p < 0.0001, OR = 2.51; p = 0.0084, OR = 2.62) and KIR2DS3 (p < 0.0001; OR = 2.57) were associated with chronic infection, independently from KIR2DL2. No differences in ancestry composition were observed between control and patients, even with respect to therapy response groups. The allelic profile KIR2DL2/KIR2DS2/KIR2DS3 was associated with the chronic hepatitis C (p < 0.0001; OR = 3). Furthermore, the patients also showed a higher mean number of activating genes and a lower frequency of the homozygous AA profile, which is likely secondary to the association with non-AA and/or activating genes. In addition, the KIR2DS5 allele was associated with SVR (p = 0.0261; OR = 0.184).The ancestry analysis of samples ruled out any effects of population substructuring and did not evidence interethnic differences in therapy response, as suggested in previous studies.

Wnt6 activates endoderm in the sea urchin gene regulatory network.

In the sea urchin, entry of ß-catenin into the nuclei of the vegetal cells at 4th and 5th cleavages is necessary for activation of the endomesoderm gene regulatory network. Beyond that, little is known about how the embryo uses maternal information to initiate specification. Here, experiments establish that of the three maternal Wnts in the egg, Wnt6 is necessary for activation of endodermal genes in the endomesoderm GRN. A small region of the vegetal cortex is shown to be necessary for activation of the endomesoderm GRN. If that cortical region of the egg is removed, addition of Wnt6 rescues endoderm. At a molecular level, the vegetal cortex region contains a localized concentration of Dishevelled (Dsh) protein, a transducer of the canonical Wnt pathway; however, Wnt6 mRNA is not similarly localized. Ectopic activation of the Wnt pathway, through the expression of an activated form of ß-catenin, of a dominant-negative variant of GSK-3ß or of Dsh itself, rescues endomesoderm specification in eggs depleted of the vegetal cortex. Knockdown experiments in whole embryos show that absence of Wnt6 produces embryos that lack endoderm, but those embryos continue to express a number of mesoderm markers. Thus, maternal Wnt6 plus a localized vegetal cortical molecule, possibly Dsh, is necessary for endoderm specification; this has been verified in two species of sea urchin. The data also show that Wnt6 is only one of what are likely to be multiple components that are necessary for activation of the entire endomesoderm gene regulatory network.

Coinfecção provocada pelos vírus da imunodeficiência humana e hepatite C (HIV-1/HCV): uma casuística da Amazônia brasileira / Coinfection caused by the human immunodeficiency virus and hepatitis C virus (HIV-1/HCV): a case study in the Brazilian Amazon

INTRODUÇÃO: Com o advento da terapia antirretroviral de alta atividade em 1996, a doença hepática tornou-se causa importante de morbidade e mortalidade em pessoas infectadas pelo vírus da imunodeficiência humana-1 (HIV-1). OBJETIVO: Descrever os aspectos demográficos e laboratoriais de 62 pacientes coinfectados com o vírus da imunodeficiência humana e da hepatite C (HIV-1/HCV). MÉTODO: Estudo transversal, incluindo pacientes portadores do HIV, confirmados sorologicamente (ELISA + Imunofluorescência indireta ou Western Blot), com anti-HCV positivos pelo teste de ELISA confirmados por RT-PCR, atendidos no ambulatório de fígado da Fundação Santa Casa de Misericórdia do Pará no período de agosto de 2004 até abril de 2008. RESULTADOS: Foram atendidos 49 (79 por cento) indivíduos do gênero masculino e 13 indivíduos do gênero feminino, com mediana de idade de 42,6 anos, solteiros (66, por cento. n = 41), heterossexuais (59,7 por cento, n = 37), bissexuais (27,4 por cento, n = 17), homem que faz sexo com homem HSH (12,9 por cento. n = 8); contagem de linfócitos T CD4+ com mediana de 327 células/mm3, carga viral plasmática do HIV com mediana de 2,54 log10 HIV-RNA cópias/mL, carga viral do HCV (HCV-RNA) de 5,90 log10 UI/mL. O genótipo 1 do HCV foi encontrado em 60,87 por cento. A biópsia hepática foi realizada em 41 (66,12 por cento) pacientes, tendo sido observada a seguinte classificação METAVIR: F0 (12 por cento), F1 (24,4 por cento), F2 (32 por cento), F3 (17 por cento) e F4 (14,6 por cento). CONCLUSÃO: Os pacientes eram predominantemente solteiros,com carga viral do HCV elevada, apresentavam fibrose de moderada a severa em mais de 50 por cento dos casos sem alterações laboratoriais significativas...

INTRODUCTION: Since highly active antiretroviral therapy was developed in 1996, liver injury has become an important cause of morbidity and mortality in individuals infected by the human immunodeficiency virus-1 (HIV-1). OBJECTIVE: To report the demographic and laboratory findings of 62 patients coinfected with HIV-1/HCV. METHODS: This cross-sectional study analyzed HIV patients, confirmed serologically by ELISA and indirect immunofluorescence or Western Blot, with positive anti-HCV by ELISA and confirmed by RT-PCR. These patiens were treated at the Liver Department of the Fundação Santa Casa de Misericórdia do Pará from August 2004 to April 2008. RESULTS: A total of 49 (79 per cent) male and 13 female patients were analyzed. Their age median was 42.6 years and they were single (66.1 per cent n=41), heterosexual (59.7 per cent, n = 37), bisexual (27.4 per cent, n = 17), man who have sex with man MSM (12.9 per cent, n = 8); the lymphocytes T CD4+ count median was 327 cells/mm3, HIV serum viral load median was 2.54 log10 HIV RNA copies/mL, HCV viral load (RNA-HCV) was 5.9 log10 UI/mL. The HCV genotype 1 was found in 60.87 per cent of the patients. Forty-one (66.12 per cent) patients were submitted to liver biopsies and the histopathology results according to METAVIR were F0 (12 per cent), F1 (24.4 per cent), F3 (17 per cent), F4 (14.6 per cent). CONCLUSION: Patients were predominantly single, with high viral load. They presented with moderate to severe fibrosis in more than 50 per cet of cases without significant changes in their laboratory findings...

Mannose-binding lectin gene polymorphisms are not associated with susceptibility to hepatitis C virus infection in the Brazilian Amazon region.

The present study compares the genotype frequencies between two population groups composed by 73 hepatitis C virus (HCV)-infected patients and 92 seronegative controls and investigates the role of allele variants as a possible factor in the susceptibility to HCV infection and the influence on disease progression. The identification of MBL*B and MBL*C alleles was performed by restriction fragment length polymorphism analysis of the 349-bp product using BanI and MboII restriction enzymes, respectively, and a polymerase chain reaction-sequence-specific polymorphism for discrimination of MBL*D. The analysis of allele and genotype frequencies between an HCV-infected group and seronegative controls did not indicate significant differences. The comparison of chronically infected subjects with and without liver cirrhosis was also not statistically significant. The odds ratio estimations were not significant, and the values obtained cannot suggest that the presence of allele variant MBL*B could have some influence in the risk of HCV infection progression to liver cirrhosis and that the presence of allele MBL*D could confer some protection against disease progression, but a larger sample size is necessary to confirm the present results.

Avaliação da resposta terapêutica de pacientes em tratamento para hepatite B crônica em hospital de referência para doenças hepáticas / Evaluation of the therapeutic response of patients in treatment for chronic hepatitis B in a hospital of reference for liver diseases

Objetivo: descrever as respostas bioquímica, sorológica e virológica à terapêutica em pacientes com hepatite B crônica em um hospital de referência para doenças hepáticas. Método: estudo transversal, analítico, onde foram coletados dados de 25 pacientes com hepatite B crônica tratados com interferon ?-2a 5MU/dia, lamivudina 100mg/dia, entecavir 1mg/dia e adefovir dipivoxil 10mg/dia. Estes pacientes foram atendidos no Ambulatório do Grupo do Fígado da Fundação Santa Casa de Misericórdia do Pará, no período de agosto de 2000 a dezembro de 2008. Os pacientes foram distribuídos em 2 grupos: HBeAg positivos e HBeAg negativos. Consideraram-se como respostas, a normalização de ALT, negativação do VHB-DNA por PCR, para os grupos de HBeAg positivos e negativos; e soroconversão HBeAg/anti-HBe, para o grupo HBeAg positivo. Resultados: observou-se que no grupo dos pacientes HBeAg positivos, houve soroconversão HBeAg/anti-HBe em 53,3%. Houve normalização de ALT em 68%, onde 44% pertencia ao grupo HBeAg positivo e 24% ao grupo HBeAg negativo. A negativação do VHB-DNA ocorreu em 68% dos pacientes, sendo 40% do grupo HBeAg positivo e 28% do grupo HBeAg negativo. Conclusão: todos os pacientes tratados com entecavir e adefovir, embora numa pequena casuística, apresentaram resposta bioquímica, sorológica e virológica. Aqueles tratados com interferon também apresentaram bons resultados. Porém, os que foram tratados com lamivudina, embora maior parte tenha apresentado resposta bioquímica e virológica favoráveis, tiveram respostas inferiores em relação aos demais esquemas terapêuticos avaliados nesta pesquisa.

Objective: to describe the therapeutic response biochemical, serologic and virologic in patients with chronic hepatitis B in a reference hospital for liver diseases. Method: cross-sectional study, analytical, where data were collected from 25 patients with chronic hepatitis B treated with interferon ?-2a 5MU/day, lamivudine 100 mg/day, entecavir 1mg/day and adefovir dipivoxil 10 mg/day. These patients were treated at the Clinic of the Liver´s Group of the Fundação Santa Casa de Misericórdia do Pará, from August 2000 to December 2008. The patients were divided into 2 groups: HBeAg-positive and HBeAg-negative. It is considered as responses, the normalization of ALT levels and undetectable HBV-DNA levels by PCR, for the groups of positive and negative HBeAg; and seroconversion HBeAg/anti-HBe, for the HBeAg-positive group. Results: it was observed that in the patients' group HBeAg-positive, there were seroconversion HBeAg/anti-HBe in 53,3%. There was normalization of ALT levels in 68%, while 44% belonged to the HBeAg positive and 24% of HBeAg negative group. Non-detectability of HBV-DNA by PCR ocurred in 68% of the patients, being 40% of the HBeAg-positive group and 28% of the HBeAg-negative group. Conclusion:. concluded that all the patients treated with entecavir and adefovir, although in a small casuistry, achieved biochemical, serologic and virologic responses. Those treated with standard interferon also showed good results. However, those who were treated with lamivudine, although larger part has obtained biochemical and virologic favorable response, they had lower results in relation to the other drugs evaluated in this study

Prevalència dos marcadores sorólogicos das hepatites virais B, C e Delta em pacientes atendidos em um Hospital de Referência de Doenças Hepáticas Crônicas do Pará / Prevalence of serologic markers of viral hepatitis B, C and Delta in patients admitted at the chronic liver diseases center of a reference hospital in Pará

Descrever a prevalência dos marcadores sorológicos das hepatites virais B, C e Delta e da coinfecção das hepatites virais B e C, além de obter informações sobre características demográficas nos pacientes atendidos no Programa de hepatopatias crônicas do hospital Fundação Santa Casa de Misericórdia do Pará. Método: foram analisados dados de 2009 pacientes com diagnóstico de hepatite crônica, que tivessem sorologia positiva para o HBsAg e/ou anti-HCV e/ou anti-HD, no período de janeirode 1996 a dezembro de 2007. Resultados: observou-se que 35,6% dos pacientes apresentaram positividade para o marcador sorológico anti-HCV, 14% dos pacientes para o HBsAg e 0,9% dos pacientes apresentaram positividade para o anti-HD. Conclusão: o estudo revelou que a maioria dos pacientes atendidos no programa de hepatopatias crônicas do Hospital Fundação Santa Casa de Misericórdia doPará apresentou o marcador sorológico para Hepatite C, pertencia ao sexo masculino, à etnia parda, era casada, enquadrava-se na faixa etária produtiva e era procedente de Belém.

To describe the prevalence of serological markers of viral hepatitis B, C and Delta and co-infection of viral hepatitis B and C as well as collect information on demographic characteristics about patients treated in the Programme of chronic hepatopathology at the Hospital Fundacão SantaCasa de Misericordia of Para. Method: for that, 2,009 records of patients diagnosed with chronic hepatitis were analyzed, which had positive serology for the HBsAg and/or anti-HCV, from January 1996 to December 2007. Results: it was observed that 35.6% of patients had positive serological marker for the anti-HCV, 14% of patients for the serological marker HBsAg and 0.9% of the patients were positive for the anti-HD. Conclusion: the study showed that the majority of patients withchronic hepatopathologies treated at service of the Hospital Fundação Santa Casa de Misericordia do Pará presented the serologic marker for Hepatitis C; they were men, had brown skin , were married, were at the productive age group and came from Belém.

LvNumb works synergistically with Notch signaling to specify non-skeletal mesoderm cells in the sea urchin embryo.

Activation of the Notch signaling pathway segregates the non-skeletogenic mesoderm (NSM) from the endomesoderm during sea urchin embryo development. Subsequently, Notch signaling helps specify the four subpopulations of NSM, and influences endoderm specification. To gain further insight into how the Notch signaling pathway is regulated during these cell specification events, we identified a sea urchin homologue of Numb (LvNumb). Previous work in other model systems showed that Numb functions as a Notch signaling pathway antagonist, possibly by mediating the endocytosis of other key Notch interacting proteins. In this study, we show that the vegetal endomesoderm expresses lvnumb during the blastula and gastrula stages, and that the protein is localized to the presumptive NSM. Injections of lvnumb mRNA and antisense morpholinos demonstrate that LvNumb is necessary for the specification of mesodermal cell types, including pigment cells, blastocoelar cells and muscle cells. Functional analysis of the N-terminal PTB domain and the C-terminal PRR domain of LvNumb shows that the PTB domain, but not the PRR domain, is sufficient to recapitulate the demonstrable function of full-length LvNumb. Experiments show that LvNumb requires an active Notch signal to function during NSM specification and that LvNumb functions in the cells responding to Delta and not in the cells presenting the Delta ligand. Furthermore, injection of mRNA encoding the intracellular domain of Notch rescues the LvNumb morpholino phenotype, suggesting that the constitutive intracellular Notch signal overcomes, or bypasses, the absence of Numb during NSM specification.

Estudo clínico, laboratorial e histopatológico de grupos selecionados de pacientes com hepatite C crônica em Belém, Pará / Clinic study, laboratory and histopathology of selected groups of patients with chronic hepatitis C in Belem, Para

A hepatite C constitui um grave problema de saúde publica no Brasil. A maioria dos dados de prevalência disponível provém de grupos específicos. Em doadores de sangue, a prevalência da infecção pelo HCV varia de 0,65 por cento (região Sul) a 3,5 por cento (região Norte). A distribuição dos genótipos do HCV no Brasil é hetrerogênea entre as diferentes regiões geográficas e, até entre diferentes estados da mesma região. Dados sobre a distribuição dos genótipos de HCV na região Norte são escassos. O principal objetivo deste estudo foi descrever aspectos clínicos, histopatológicos e laboratoriais de diferentes categorias de pacientes com hepatite C crônica, atendidos em um centro de referência para atendimento e tratamento de pacientes com doenças hepática crônica, em Belém, Pará. Um estudo de corte transversal foi conduzido em 345 indivíduos infectados pelo HCV (163 pacientes com hepatite crônica, 56 doadores de sangue, 32 hemofílicos, 73 hemodialisados e 22 pacientes com carcinoma hepatocelular), procedentes de diferentes municípios do estado do Pará. Foram avaliados os seguintes aspectos demográficos, clínicos e epidemiológicos: idade, sexo, apresentação clínica, fatores de risco de aquisição de infecção por HCV e grau de lesões hepáticas de acordo com o sistema de classificação METAVIR. Amostras de soro foram analisadas para determinação do RNA e dos genótipos do HCV. A média de idade dos 345 pacientes com hepatite C crônica foi 47,7 anos (mais ou menos 13,9). A maioria dos pacientes, neste estudo, era do sexo masculino (74,8 por cento) e as mulheres apresentaram média de idade mais elevada (p=0.00115) do que os homens (51,9 mais ou menos 12,9 anos e 48,6 mais ou menos 13,7 anos, respectivamente). Os fatores de risco associados à infecção por HCV mais freqüentes foram: história prévia de transfusão sanguínea (50,1 por cento), cirurgia (36,4 por cento), hemodiálise (31,2 por cento) e compartilhamento de agulhas não descartáveis (16,8 por cento). As manifestações extra-hepáticas mais freqüentes nestes pacientes foram: astenia, hipertensão, diabetes mellitus, artralgia, prurido, hipotireoidismo e trombocitopenia. Da população estudada, 98 por cento dos pacientes estavam infectados pelo genótipo 1 (77,1 por cento) ou 3 (20,9 por cento) do HCV. A distribuição dos genótipos do HCV foi similar entre os cinco grupos de pacientes. Não foram observadas diferenças significativas entre os genótipos do HCV e fatores de risco, aspectos clínicos e demográficos, exceto idade. Indivíduos infectados pelo HCV genótipo 3 apresentaram média de idade mais baixa (44,5 mais ou menos 13,6 anos) quando comparados àqueles infectados pelo HCV genótipo 1 (48,6 mais ou menos 13,7 anos) (p= 0.0273). Os seguintes aspectos clínicos e histopatológicos parecem estar associados com o genótipo de HCV: maior prevalência de diabetes mellitus tipo 2 (p= 0,0425) nos pacientes infectados pelo genótipo. A intensidade da fibrose foi associada a idade elevada, grau de atividade e esteatose, níveis séricos elevados de ALT e AST e plaquetopenia. A presença de esteatose foi associada a graus de atividade e fibrose. Os maiores percentuais de resposta virológica sustentada (RVS) ao tratamento foram observados nos pacientes tratados com Interferon-alfa peguilado combinado com ribavirina.