RESUMEN

The present article provides an improvement in the method to correct indirect strain measurements in triaxial compressive strength tests through axial displacement and hydraulic fluid volume change measurements. The improvement focused on reducing the parameters of the formula proposed for indirect volumetric strain in the original method, thereby facilitating the development of a simpler formula in which the radial strain depends on only two parameters: the initial volume of the rock specimen and the volume changes of the hydraulic fluid for each instant. The comparison between the improvement proposed, and original method resulted in a mean absolute difference of 0.003.•This improvement does not depend on the axial strain, unlike the original method, which requires correcting the indirect axial strain measurements before correcting the indirect radial strain measurements.•This improvement can be useful for research on the stress-strain behavior of intact rock under laboratory conditions, such as in the study of the post-peak state.

RESUMEN

Dealing with electromyography (EMG) signals is often not simple. The nature of these signals is nonstationary, noisy, and high dimensional. These EMG characteristics make their predictability even more challenging. Cross recurrence plots (CRPs) have demonstrated in many works their capability of detecting very subtle patterns in signals often buried in a noisy environment. In this contribution, fifty subjects performed ten different hand movements with each hand with the aid of electrodes placed in each arm. Furthermore, the nonlinear features of each subject's signals using cross recurrence quantification analysis (CRQA) have been performed. Also, a novel methodology is proposed using CRQA as the mainstream technique to detect and classify each of the movements presented in this study. Additional tools were presented to determine to which extent this proposed methodology is able to avoid false classifications, thus demonstrating that this methodology is feasible to classify surface EMG (SEMG) signals with good accuracy, sensitivity, and specificity. Lastly, the results were compared with traditional machine learning methods, and the advantages of using the proposed methodology above such methods are highlighted.

Asunto(s)

Electromiografía/estadística & datos numéricos , Mano/fisiología , Adolescente , Adulto , Biología Computacional , Femenino , Humanos , Aprendizaje Automático , Masculino , Persona de Mediana Edad , Modelos Biológicos , Movimiento/fisiología , Dinámicas no Lineales , Procesamiento de Señales Asistido por Computador , Relación Señal-Ruido , Análisis de Ondículas , Adulto Joven

RESUMEN

PURPOSE: The third most frequently diagnosed cancer in Europe in 2018 was lung cancer; it is also the leading cause of cancer death in Europe. We studied patient and tumor characteristics, and patterns of healthcare provision explaining regional variability in lung cancer survival in southern Spain. METHODS: A population-based cohort study included all 1196 incident first invasive primary lung cancer (C33-C34 according to ICD-10) cases diagnosed between 2010 and 2011 with follow-up until April 2015. Data were drawn from local population-based cancer registries and patients' hospital medical records from all public and private hospitals from two regions in southern Spain. RESULTS: There was evidence of regional differences in lung cancer late diagnosis (58% stage IV in Granada vs. 65% in Huelva, p value < 0.001). Among patients with stage I, only 67% received surgery compared with 0.6% of patients with stage IV. Patients treated with a combination of radiotherapy, chemotherapy, and surgery had a 2-year mortality risk reduction of 94% compared with patients who did not receive any treatment (excess mortality risk 0.06; 95% CI 0.02-0.16). Geographical differences in survival were observed between the two regions: 35% vs. 26% at 1-year since diagnosis. CONCLUSIONS: The observed geographic differences in survival between regions are due in part to the late cancer diagnosis which determines the use of less effective therapeutic options. Results from our study justify the need for promoting lung cancer early detection strategies and the harmonization of the best practice in lung cancer management and treatment.

Asunto(s)

Detección Precoz del Cáncer/mortalidad , Servicios de Salud , Disparidades en Atención de Salud , Neoplasias Pulmonares/mortalidad , Sistema de Registros/estadística & datos numéricos , Adolescente , Adulto , Anciano , Estudios de Cohortes , Terapia Combinada , Femenino , Estudios de Seguimiento , Humanos , Neoplasias Pulmonares/epidemiología , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/terapia , Masculino , Persona de Mediana Edad , Pronóstico , Factores de Riesgo , España/epidemiología , Tasa de Supervivencia , Adulto Joven

RESUMEN

HMGB-like proteins are architectural chromatin factors, and their function is heavily dependent on their ability to interact with DNA (especially non-canonical DNA structures). HMGB1 is involved in many DNA processes, and dysregulation of HMGB protein expression has profound effects on cellular transcription, resulting in severe developmental defects as well as cancer. During DNA replication, elements that form the origin are still not well defined in metazoans. Sites with A (adenine) or T (thymine) repeats cause intrinsic curvatures in the DNA and are described to be involved in the replication machinery by providing binding sites to replication proteins. As a result, the DNA molecule shows intrinsically bent DNA sites, caused by periodic repeats of 2 or more As/Ts (dA/dT) as well as intrinsically non-bent DNA sites (INBDs), due to a succession of curvatures that cancel each other. In the present study, we mapped 11 INBDSs present in the AMPD2 gene that are related to each replication origin (oriGNAI3, oriC, oriB, and oriA). Following characterization of INBDSs, we tested the ability of HMGB1 to bind to the bent (b1, b2, b4a, b4b, b5, b6, b7, and b8) and non-bent DNA fragments (nb7, nb11, nb1, nb2, nb4, and nb5) via electrophoretic mobility shift assays. All fragments showed efficient binding to HMGB1. However, the non-bent DNA fragments nb2, nb4, and nb5 showed slightly reduced binding efficiency.

Asunto(s)

AMP Desaminasa/genética , Replicación del ADN/genética , Proteínas de Unión al ADN/genética , Proteína HMGB1/genética , AMP Desaminasa/química , Animales , Sitios de Unión , Cromatina/química , Cromatina/genética , Cricetulus/genética , ADN/química , ADN/genética , Proteínas de Unión al ADN/química , Proteína HMGB1/química , Conformación de Ácido Nucleico , Unión Proteica , Origen de Réplica/genética

RESUMEN

In insects, ribosomal genes are usually detected in sex chromosomes, but have also or only been detected in autosomal chromosomes in some cases. Previous results from our research group indicated that in Bradysia hygida, nucleolus organizer regions were associated with heterochromatic regions of the autosomal C chromosome, using the silver impregnation technique. The present study confirmed this location of the ribosomal genes using fluorescence in situ hybridization analysis. This analysis also revealed the partial sequences of the 18S and 28S genes for this sciarid. The sequence alignment showed that the 18S gene has 98% identity to Corydalus armatus and 91% identity to Drosophila persimilis and Drosophila melanogaster. The partial sequence analysis of the 28S gene showed 95% identity with Bradysia amoena and 93% identity with Schwenckfeldina sp. These results confirmed the location of ribosomal genes of B. hygida in an autosomal chromosome, and the partial sequence analysis of the 18S and 28S genes demonstrated a high percentage of identity among several insect ribosomal genes.

Asunto(s)

Mapeo Cromosómico , Dípteros/genética , Genes de Insecto , ARN Ribosómico 18S/genética , ARN Ribosómico 28S/genética , Animales , Secuencia de Bases , Hibridación Fluorescente in Situ , Datos de Secuencia Molecular , Cromosomas Politénicos , Alineación de Secuencia , Análisis de Secuencia de ADN

RESUMEN

Identification of the nucleotide consensus sequence in mammalian replication origins is a difficult and controversial problem. The hypothesis that local DNA topology could be involved in recognition by replication proteins is an exciting possibility. Secondary DNA structures, including intrinsically bent DNA, can be easily detected, and they may indicate a specific pattern in or near mammalian replication origins. This work presents the entire mapping of the intrinsically bent DNA sites (IBDSs), using in silico analysis and a circular permutation assay, of the DNA replication origins oriGNAI3, oriC, oriB, and oriA in the mammalian amplified AMPD2 gene domain. The results show that each origin presents an IBDS that flanks the straight core of these DNA replication sites. In addition, the in silico prediction of the nucleosome positioning reveals a strong indication that the center of an IBDS is localized in a nucleosome-free region (NFR). The structure of each of these curved sites is presented together with their helical parameters and topology. Together, the data that we present here indicate that the oriGNAI3 origin where preferential firing to the replication initiation events in the amplified AMPD2 domain occurs is the only origin that presents a straight, narrow region that is flanked on both sides by two intrinsically bent DNA sites within a short distance (~300 bp); however, all of the origins present at least one IBDS, which is localized in the NFR region. These results indicate that structural features could be implicated in the mammalian DNA replication origin and support the possibility of detecting and characterizing these segments.

Asunto(s)

AMP Desaminasa/genética , ADN/química , AMP Desaminasa/metabolismo , Animales , Secuencia de Bases , ADN/metabolismo , Sitios Genéticos , Datos de Secuencia Molecular , Regiones Promotoras Genéticas , Origen de Réplica/genética

RESUMEN

The domesticated silkworm Bombyx mori L. is currently found only in germplasm banks. Therefore, characterization and conservation of this genetic resource is crucial. Based on previous studies that revealed nucleotide differences in silkworm strains, the intron of the fibroin heavy chain gene (H-fib) can be used for molecular silkworm characterization. The H-fib gene has two exons and a unique intron, and encodes the principal component of the silk fiber, the fibroin heavy chain. Therefore, this study aimed to identify the genetic variability of the unique intron of H-fib gene of 20 silkworm strains maintained at the Universidade Estadual de Maringá Brazilian Germplasm Bank (UBGB) by conformation-sensitive gel electrophoresis (CSGE) and nucleotide sequencing. Genomic DNA extracted from silkworm moths was PCR amplified. CSGE revealed that most of the analyzed silkworm strains had only homoduplex molecules. However, DNA from the Japanese strains B106, B82, and M12-2 had two extra DNA fragments produced by heteroduplex molecules, revealing variation between alleles. Sequencing of the H-fib intron was used to confirm the variation previously detected by CSGE and detected a significant polymorphism characterized by a 17-base pair (bp) deletion, a 2-bp insertion, and eight nucleotide substitutions. Although genetic and allelic variability was detected in some silkworm strains, the intron of the H-fib gene revealed not to be the best molecular marker for the characterization of B. mori strains from UBGB.

Asunto(s)

Bombyx/genética , Genes de Insecto , Alelos , Animales , Brasil , Fibroínas , Intrones , Polimorfismo Genético

RESUMEN

Brazil has only one public genetic pool of Bombyx mori strains, which was established in 2005 at Universidade Estadual de Maringá, Maringá, Paraná State. This genetic bank has been maintained, and the strains have been characterized using genetic and morphological tools. The quantitative and qualitative traits, directly or indirectly related to productivity, were evaluated in 14 silkworm strains. In addition to biological and productivity analyses, DNA markers related to susceptibility to the B. mori nucleopolyhedrovirus (BmNPV) were analyzed. BmNPV is a major cause of production loss and is a serious problem for Paraná sericulture. The silkworm strains from diverse geographic origins were found to have different characteristics, including body weight, larval stage duration, cocoon weight, and other biological traits. In terms of productivity, the raw silk percentages were almost uniform, with an overall average of 16.28%. Overall, the Chinese strain C37 gave the best performance in many of the quantitative traits, and it surpassed the other strains in productivity traits. Therefore, it can be used as one of the strains that compose the elite germplasm for silkworm breeding programs. Additionally, genetic molecular markers were efficient in discriminating between B. mori strains that had been identified based on their geographical origin. We found that all Japanese strains produced a 400-bp molecular marker that has been associated with susceptibility to BmNPV.

Asunto(s)

Bombyx/genética , Bombyx/fisiología , Marcadores Genéticos , Nucleopoliedrovirus/fisiología , Animales , Bancos de Muestras Biológicas , Bombyx/anatomía & histología , Bombyx/virología , Brasil , Cruzamiento , Bases de Datos Genéticas , Larva/anatomía & histología , Larva/genética , Larva/fisiología , Carácter Cuantitativo Heredable , Seda , Textiles

RESUMEN

This study examined the phylogenetic relationship of strains of canine distemper virus (CDV) collected from Paraná State, Brazil, based on the hemagglutinin gene. Urine samples were collected from 4 dogs from northern Paraná State that demonstrated clinical manifestations of canine distemper. The participation of CDV was initially confirmed by RT-PCR targeting the nucleocapsid protein, after which the complete hemagglutinin gene was sequenced from each sample. Sequences were deposited in and compared with those already in GenBank. Phylogenetic analyses, using amino acid and nucleotide sequences based on the hemagglutinin gene, demonstrated that these strains of CDV are closely related to those from the Europe 1 lineage of CDV, with marked differences from other recognized geographical clusters of CDV isolates and from the vaccine strains. The strains of CDV from this region of southern Brazil appear to be related to those from Europe 1.

Asunto(s)

Virus del Moquillo Canino/genética , Genes Virales , Hemaglutininas Virales/genética , Filogenia , Brasil

RESUMEN

Imposex, a syndrome characterized by the appearance of a penis and/or vas deferens in female gastropods due the presence of organotins in environment, is still observed in Brazilian gastropod populations, as in other countries, even after the tributyltin (TBT) ban. Nevertheless, the progressive controls on the use of organotins in antifouling systems at national and international levels and the consequent reduction of their environmental concentrations have led to changes in the characteristics of imposex development observed in Stramonita haemastoma. Populations of this species were analyzed on the coast of Rio de Janeiro (Brazil) between 2007 and 2008, and the developmental pathways associated with the syndrome were identified. Compared with previous works, it was noted that imposex expression was reduced in most of Guanabara Bay. Aphallic imposex development, on the other hand, showed a marked increase. The pathways of imposex development were also evaluated in a temporal data series from a fixed sampling station at Vermelha beach, and the incidence of aphallic imposex development was found to show a marked increase from 1998 onward. Furthermore, the observation of either the presence or absence of a penis in imposex-affected females may indicate that penis development is related to the contaminant exposure level and that the decreasing TBT concentrations in the local environment result in the predominance of an aphallic route of imposex development. These findings support the idea that imposex female aphally in this species could be a dose-dependent response, rather than a genetic anomaly.

Asunto(s)

Gastrópodos/efectos de los fármacos , Compuestos Orgánicos de Estaño/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Brasil , Femenino , Masculino , Compuestos de Trialquiltina/toxicidad , Conducto Deferente/efectos de los fármacos

RESUMEN

This study evaluated the effectiveness of a disinfectant with low corrosive action and which is not toxic to the environment, the sodium dichloroisocyanurate formulation, on the Bombyx mori nucleopolyhedrovirus (BmNPV). For this, 5th-instar B. mori silkworm larvae were divided into four experimental groups of 4 replicates with 15 larvae each, totalling 60 larvae per group. The groups were fed with mulberry (Morus sp.) leaf discs containing: BmNPV treated with the disinfectant, untreated BmNPV, only the disinfectant, and water (control). The results showed that the disinfectant does not inactivate the BmNPV and also exerts a negative effect on the insect"s resistance.(AU)

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O estudo avaliou a eficiência de um desinfetante que apresenta baixa ação corrosiva e que não é tóxico ao meio ambiente, o formulado de sódio dicloroisocianurato, sobre o nucleopoliedrovírus Bombyx mori (BmNPV). Para tanto, lagartas do bicho-da-seda, B. mori, de 5º instar foram divididas em quatro grupos experimentais, 4 repetições com 15 lagartas cada, totalizando 60 lagartas por grupo. Os grupos foram alimentados com discos foliares de amoreira (Morus sp.) contendo: o BmNPV tratado com o desinfetante (solução 1); o BmNPV não tratado (solução 2); apenas o desinfetante (solução 3); e água (solução 4, controle). Os resultados mostraram que o desinfetante não inativa o BmNPV e também exerce efeito negativo na resistência do inseto.(AU)

Asunto(s)

Animales , Desinfección , Desinfectantes , Baculoviridae , Bombyx , Toxicidad

RESUMEN

This study evaluated the effectiveness of a disinfectant with low corrosive action and which is not toxic to the environment, the sodium dichloroisocyanurate formulation, on the Bombyx mori nucleopolyhedrovirus (BmNPV). For this, 5th-instar B. mori silkworm larvae were divided into four experimental groups of 4 replicates with 15 larvae each, totalling 60 larvae per group. The groups were fed with mulberry (Morus sp.) leaf discs containing: BmNPV treated with the disinfectant, untreated BmNPV, only the disinfectant, and water (control). The results showed that the disinfectant does not inactivate the BmNPV and also exerts a negative effect on the insect"s resistance.

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O estudo avaliou a eficiência de um desinfetante que apresenta baixa ação corrosiva e que não é tóxico ao meio ambiente, o formulado de sódio dicloroisocianurato, sobre o nucleopoliedrovírus Bombyx mori (BmNPV). Para tanto, lagartas do bicho-da-seda, B. mori, de 5º instar foram divididas em quatro grupos experimentais, 4 repetições com 15 lagartas cada, totalizando 60 lagartas por grupo. Os grupos foram alimentados com discos foliares de amoreira (Morus sp.) contendo: o BmNPV tratado com o desinfetante (solução 1); o BmNPV não tratado (solução 2); apenas o desinfetante (solução 3); e água (solução 4, controle). Os resultados mostraram que o desinfetante não inativa o BmNPV e também exerce efeito negativo na resistência do inseto.

Asunto(s)

Animales , Baculoviridae , Bombyx , Desinfección , Desinfectantes , Toxicidad

RESUMEN

Molecular combing technology is an important new tool for the functional and physical mapping of genome segments. It is designed to identify amplifications, microdeletions, and rearrangements in a DNA sequence and to study the process of DNA replication. This technique has recently been used to identify and analyze the dynamics of replication in amplified domains. In Bradysia hygida, multiple amplification initiation sites are predicted to exist upstream of the BhC4-1 gene. However, it has been impossible to identify them using the available standard techniques. The aim of this study was to optimize molecular combing technology to obtain DNA fibers from the polytene nuclei of the salivary glands of B. hygida to study the dynamics of DNA replication in this organism. Our results suggest that combing this DNA without prior purification of the polytene nuclei is possible. The density, integrity, and linearity of the DNA fibers were analyzed, fibers 50 to 300 kb in length were detected, and a 9-kb fragment within the amplified region was visualized using biotin detected by Alexa Fluor 488-conjugated streptavidin technique. The feasibility of physically mapping these fibers demonstrated in this study suggests that molecular combing may be used to identify the replication origin of the BhC4-1 amplicon.

Asunto(s)

Replicación del ADN/genética , Dípteros/crecimiento & desarrollo , Dípteros/genética , Amplificación de Genes/genética , Regulación del Desarrollo de la Expresión Génica , Mapeo Físico de Cromosoma/métodos , Animales , Humanos , Análisis de Secuencia por Matrices de Oligonucleótidos , Glándulas Salivales/metabolismo

RESUMEN

Previous reports demonstrated that actin is necessary for nucleocapsid transport and viral gene expression during nucleopolyhedrovirus infection of Bombyx mori. The first intron of B. mori A3 actin contains a cryptic promoter that drives expression of a rare isoform. We detected differences in the size and nucleotide composition of the first intron of the A3 actin gene from B. mori strain C24A, which is more resistant to nucleopolyhedrovirus than the M11A strain (22 and 95% lethality, respectively). We sought to determine if resistance to BmMNPV infection and the A3 actin promoter structure are correlated. Intrinsically bent DNA sites in these sequences, which determine curved structures, were analyzed by electrophoretic mobility assays and the helical parameters ENDS ratio, roll and twist. We found both fragments to have non-centralized bent DNA sites with distinct ENDS ratio values, nucleotide positions and two-dimensional structures. Additionally, a conformational-sensitive gel electrophoresis assay identified an allelic variation found in strain M11A that is absent in strain C24A. These data suggest that A3 actin intronic sequence variations impair virus propagation and are markers of BmMNPV-resistant populations.

Asunto(s)

Actinas/genética , Bombyx/genética , Proteínas de Insectos/genética , Intrones , Nucleopoliedrovirus/fisiología , Regiones Promotoras Genéticas , Animales , Bombyx/virología , Larva/genética , Datos de Secuencia Molecular , Alineación de Secuencia

RESUMEN

A nuclear polyhedrosis virus isolated from infected Bombyx mori, BmNPV, was used to inoculate silkworms to determine salivary gland cell susceptibility. The salivary gland was removed from infected silkworms at different times post-inoculation and examined by light microscopy. The salivary gland cells did not exhibit any signs of BmNPV infection; however, fat body and tracheal cells, used as positive controls, showed characteristic cytopathological changes caused by BmNPV infection, which confirmed inoculum viability. The morphological distribution of tracheal branches and the basal lamina, which serves as a barrier to viral penetration, are apparently involved in this resistance to infection.

Asunto(s)

Bombyx/virología , Nucleopoliedrovirus/fisiología , Glándulas Salivales/virología , Animales , Cuerpo Adiposo/virología

RESUMEN

Diatraea saccharalis is an insect that causes considerable losses in the sugar cane crop. Our aim was to contribute to the knowledge of the biology of D. saccharalis, with the report of DNA fragments involved in the differentiation between the male and female of this species using the RAPD sex molecular marker GyakuU-13, which is specific for the W chromosome of Bombyx mori. Another point evaluated in this study was the genetic diversity of a D. saccharalis population maintained by inbreeding in a laboratory culture. The profile of sex-specific fragments was analyzed, and the genetic variability of this population was estimated. An analysis of the molecular markers showed only one fragment, of approximately 700 bp, that could be considered as a female sex marker in D. saccharalis.

Asunto(s)

Variación Genética , Mariposas Nocturnas/genética , Técnica del ADN Polimorfo Amplificado Aleatorio , Animales , Secuencia de Bases , Análisis por Conglomerados , Cartilla de ADN , Femenino , Masculino

RESUMEN

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80% identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Asunto(s)

Antibacterianos/farmacología , Hemolinfa/química , Lepidópteros/química , Péptidos/farmacología , Proteínas/análisis , Animales , Antibacterianos/aislamiento & purificación , Péptidos y Proteínas de Señalización Intercelular , Larva/química , Espectrometría de Masas , Péptidos/aislamiento & purificación , Proteínas/metabolismo , Sepsis/metabolismo

RESUMEN

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80 percent identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Asunto(s)

Animales , Antibacterianos/farmacología , Hemolinfa/química , Lepidópteros/química , Péptidos/farmacología , Proteínas/análisis , Antibacterianos/aislamiento & purificación , Larva/química , Espectrometría de Masas , Péptidos/aislamiento & purificación , Proteínas/metabolismo , Sepsis/metabolismo

RESUMEN

ORFs 10 and 14 from Bombyx mori multiple nucleopolyhedrovirus (BmMNPV) were amplified, cloned and sequenced. Nucleotide analysis of these genes and those of other baculoviruses showed that these genes are highly conserved. The p10 protein from BmMNPV ORF10 has 70 amino acid residues similar to that of the four other known BmNPV strains. The BmMNPV ORF14 alignment showed a higher identity with the nucleopolyhedrovirus ORF14 from the baculovirus BmNPV and from Autographa californica multiple nucleopolyhedrovirus. The BmMNPV ORF14 protein has a putative transmembrane domain in the C-terminal region, which is similar to that of other baculoviruses. A phylogenetic analysis showed that BmMNPV ORF14 protein has higher similarity with BmNPV ORF14 and ORF23 of A. californica multicapsid nucleopolyhedrovirus (Ac23). We conclude that proteins produced by ORFs 10 and 14 from BmNPV and BmMNPV are highly conserved in NPVs and MNPVs. The high degree of conservation among members of these genera indicates the importance of these proteins, which could mean an important function that is active throughout the infection cycle.

Asunto(s)

Bombyx/virología , Secuencia Conservada , Nucleopoliedrovirus/genética , Sistemas de Lectura Abierta/genética , Secuencia de Aminoácidos , Animales , Datos de Secuencia Molecular , Filogenia , Estructura Terciaria de Proteína , Alineación de Secuencia , Análisis de Secuencia de ADN , Análisis de Secuencia de Proteína , Proteínas Virales/química , Proteínas Virales/genética

RESUMEN

The human cases of eosinophilic meningitis recently reported from Brazil have focused the attention of the public health agencies on the role the introduced snail Achatina fulica plays as hosts of the metastrongylid nematodes. Determining the potential of this snail to host and develop infective larval stages of metastrongylids in the wild and identify the species harbored by them is crucial for designing effective control measures. Here we assess if A. fulica may act as intermediate host of A. cantonensis at the peridomiciliary areas of a patient's house from state of Pernambuco (PE), who was diagnosed with eosinophilic meningitis and a history of ingesting raw molluscs. Larvae obtained from naturally infected A. fulica were orally administered to Rattus norvegicus. The worms were collected from the pulmonary artery and brain, and were morphologically characterized and compared to the Japan isolate of A. cantonensis. Adult worms and infective L(3) larvae (PE isolate) recovered from A. fulica specimens were also analyzed by polymerase chain reaction and restriction fragment length polymorphism of ITS2 region from rDNA and compared to A. cantonensis (ES isolate), A. vasorum (MG isolate) and A. costaricensis (RS isolate). The large size of the spicules (greater than those observed in other species of Angiostrongylus) and the pattern of the bursal rays agree with the original species description by Chen (1935). Furthermore, the morphology of the PE isolate was similar to that of Japan isolate. The PCR-RFLP profiles obtained were distinctive among species and no variation in patterns was detected among adult individuals from A. cantonensis isolates from PE and ES. The importance of A. fulica as an intermediate host of eosinophilic menigoencepahlitis in Brazil is emphasized.

Asunto(s)

Angiostrongylus cantonensis/aislamiento & purificación , Caracoles/parasitología , Adulto , Angiostrongylus cantonensis/anatomía & histología , Angiostrongylus cantonensis/crecimiento & desarrollo , Angiostrongylus cantonensis/patogenicidad , Animales , Encéfalo/parasitología , Brasil , Niño , ADN de Helmintos/genética , ADN Ribosómico/genética , ADN Espaciador Ribosómico/genética , Femenino , Genotipo , Humanos , Estadios del Ciclo de Vida , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Arteria Pulmonar/parasitología , Ratas , Infecciones por Strongylida/parasitología , Infecciones por Strongylida/patología