Relato de caso: linfoma de células t/nk extranodal tipo nasal / Case Report: Extranodal Nk/T-Cell Lymphoma, N a s a l Ty p e / Reporte de caso: linfoma extranodal de células t/nk de tipo nasal

Introdução: O linfoma não-Hodgkin é dividido em linfomas de células B e linfomas de células T, e o linfoma extranodal de células T / NK do tipo nasal está dentro do último grupo.Relato de caso: Paciente do sexo masculino de 30 anos, relata que há 6 meses, de forma progressiva e de início insidioso, apresenta tumor cervical à direita de crescimento progressivo, pelo qual foi encaminhado ao ambulatório de cabeça e pescoço onde apresentou seus principais sinais e sintomas adenopatia cervical direita, sintoma B e tumoração ao nível da nasofaringe, envolvendo o teto, parede posterior e face lateral; se movimenta com auxílio, com extenso conglomerado linfonodal supraclavicular direito, eritematoso, com calor local, além de áreas de ulceração e secreção serosa.Conclusão: O diagnóstico e tratamento precoces desta doença são as únicas ferramentas para melhorar o mau prognóstico e o grave impacto na qualidade de vida dos pacientes que a padecem

Introduction: Non-Hodgkin's lymphomas are divided into B-cell lymphomas and T-cell lymphomas, and extranodal NK/T-cell lymphoma, nasal type, is in the latter group.Case report: A 30-year-old male patient, for six months, progressively and with an insidious onset, has had a right-sided cervical tumor with progressive growth. He came to a head and neck outpatient clinic where the main signs and symptoms detected were right cervical lymphadenopathy, B-symptoms, and a tumor in the nasopharynx affecting the roof, posterior wall, and lateral wall. The patient moves with assistance and has an enlarged, erythematous warm right supraclavicular lymph node conglomerate. In addition, he has some ulcerated areas with serous drainage.Conclusion: Early diagnosis and treatment of this disease are the only tools to improve these patients' poor prognosis and severely deteriorated quality of life.

Introducción:El linfoma no Hodgkin se divide en linfomas de células B y linfomas de células T; y en este último grupo se encuentra el linfoma extraganglionar de células T / NK de tipo nasal.Caso clínico: Un paciente masculino de 30 años refiere que durante 6 meses de forma progresiva, y con un início insidioso, presenta una tumoración cervical en el lado derecho de crecimiento progresivo, por lo que acude a la consulta externa de cabeza y cuello, donde los signos y síntomas principales fueron adenopatía cervical derecha, síntoma B, y una tumoración a nivel de nasofaringe, que afecta el techo, la pared posterior y la cara lateral. Se moviliza con ayuda, con un extenso conglomerado ganglionar supraclavicular derecho, eritematoso, con calor local. Además, también muestra algunas áreas de ulceración y secreción serosa. Conclusión: El diagnóstico y tratamiento precoz de esta enfermedad son las únicas herramientas para mejorar el mal pronóstico y el deterioro severo en la calidad de vida de los pacientes que la padecen

A Cell Line Adapted Infectious Laryngotracheitis Virus Strain (BΔORFC) for in ovo and Hatchery Spray Vaccination Alone or in Combination with a Recombinant HVT-LT Vaccine.

To produce more-stable, live attenuated vaccines for infectious laryngotracheitis virus (ILTV), deletion of genes related to virulence has been extensively pursued. Although its function remains unknown, the open reading frame C (ORF C) is among the genes potentially associated with viral virulence that is nonessential for replication in vitro. Earlier results indicated that the ILT virus with deletion of the ORF C gene (BΔORFC) was suitable and safe for eye drop administration but was not sufficiently attenuated for in ovo administration. The objective of this study was to evaluate the safety and protection efficacy of a cell line-adapted, gene-deleted strain (BΔORFC) of ILTV when administered in ovo and/or spray (SP) by itself, or in combination with the recombinant HVT-LT (rHVT-LT) vaccine. Results indicated that vaccination with the BΔORFC strain, either by itself or in combination with an rHVT-LT vaccine, did not affect hatchability, and only marginal signs of respiratory distress were recorded for groups of chickens that received the BΔORFC strain via SP. The replication and seroconversion induced by the BΔORFC strain after in ovo and SP administration was very limited, whereas the replication of the rHVT-LT vaccine was delayed when combined with the BΔORFC strain in ovo. Compared to rHVT-LT or BΔORFC when administered alone, dual vaccination with rHVT-LT + BΔORFC was more effective in mitigating clinical signs of the disease and reducing challenge virus load in the trachea. To our knowledge, this study provides the first proof of concept that ILTV strains can be sufficiently attenuated for early vaccination in ovo or at hatch; also, this study documented the benefits of using a dual (recombinant and live attenuated) hatchery vaccination strategy for ILTV.

Una cepa del virus de la laringotraqueítis infecciosa adaptada a una línea celular (BΔORFC) para vacunación in ovo y en aerosol en incubadora aplicada por sí sola o en combinación con una vacuna recombinante para laringotraqueítis con el vector HVT. Para producir vacunas vivas atenuadas más estables contra el virus de la laringotraqueítis infecciosa (ILTV), se ha buscado ampliamente la eliminación de genes relacionados con la virulencia. Aunque su función sigue siendo desconocida, el marco de lectura continuo C (ORF C) se encuentra entre los genes potencialmente asociados con la virulencia viral que no es esencial para la replicación in vitro. Resultados anteriores indican que el virus de la laringotraqueítis infecciosa con deleción del gene ORF C (BΔORFC) era adecuado y seguro para la administración ocular, pero no estaba lo suficientemente atenuado para su administración in ovo. El objetivo de este estudio fue evaluar la seguridad y la eficacia de la protección de una cepa del virus de la laringotraqueítis infecciosa con deleción genética y adaptada a una línea celular (BΔORFC) cuando se administra in ovo y/o en aerosol por sí sola, o en combinación con una vacuna recombinante con el vector HVT (vacuna rHVT-LT). Los resultados indicaron que la vacunación con la cepa BΔORFC, ya sea sola o en combinación con la vacuna rHVT-LT, no afectó la incubabilidad, y solo se registraron signos marginales de dificultad respiratoria para los grupos de pollos que recibieron la cepa BΔORFC por aspersión. La replicación y seroconversión inducida por la cepa BΔORFC después de la administración in ovo y por aspersión fue muy limitada, mientras que la replicación de la vacuna rHVT-LT se retrasó cuando se combinó con la cepa BΔORFC in ovo. En comparación con las vacunas rHVT-LT o BΔORFC administradas por sí solas, la vacunación dual con rHVT-LT + BΔORFC fue más eficaz para mitigar los signos clínicos de la enfermedad y reducir la carga del virus de desafío en la tráquea. Hasta donde se conoce, este estudio proporciona la primera prueba del concepto de que las cepas del virus de la laringotraqueítis infecciosa pueden atenuarse lo suficiente para la vacunación temprana in ovo o en la incubadora. Además, este estudio documentó los beneficios de utilizar una estrategia de vacunación de incubadora dual (vacuna recombinante y viva atenuada) para la laringotraqueítis infecciosa.

Histopathologic Lesion Scoring and Histomorphometric Methods for Measuring Vaccine Reactions in the Trachea of Broiler Chickens.

Severity of the tracheal histologic inflammatory response induced in broilers by ocular inoculation of two infectious bronchitis (IBV) and three Newcastle disease virus (NDV) commercial vaccines were evaluated. The vaccine was delivered by eye drop with a coarse spray to day-old chicks. The vaccines were given individually or in various combinations and were evaluated relative to nonvaccinated controls. Evaluations were performed on postvaccination (PV) days 7 and 14. Histologic endpoints included semiquantitative severity scoring of inflammatory components and quantitative morphometric determinations of inflammatory cell concentration, mucosal thickness, and percentage of ciliated mucosal surface. Strong positive correlations were observed between routine severity scoring and morphometric inflammatory parameters, whereas a negative correlation was present between inflammation severity and the percentage of mucosal ciliation. Variable, sometimes extensive, and often statistically significant differences in inflammatory responses were observed between the various vaccines. One IBV Massachusetts strain vaccine (IBV-A) produced the greatest overall inflammatory response when given alone or in combination with the NDV vaccines. Enhancement of tracheitis was seen on PV day 14 by covaccination of IBV-A with the NDV vaccines, but not by covaccination of another IBV Massachusetts strain vaccine (IBV-B) with NDV. Reduction in cilia percentage was observed for all vaccine groups relative to controls on PV day 7. However, although reactive cilia regeneration occurred on PV day 14 for most vaccine groups, a cilia regenerative response was not apparent for individual or NDV combination vaccination for IBV-A. The study also demonstrates that substantial microscopic trachea pathology may be present in vaccinated birds not exhibiting apparent clinical respiratory signs.

Artículo regular­Métodos de calificación de lesiones histológicas e histomorfométricos para medir las reacciones a las vacunas en la tráquea de pollos de engorde. Se evaluó la gravedad de la respuesta inflamatoria histológica traqueal en pollos de engorde inducida mediante la inoculación ocular de dos vacunas comerciales contra la bronquitis infecciosa (IBV) y tres vacunas del virus de la enfermedad de Newcastle (NDV). Las vacunas se administraron mediante aplicación ocular a pollitos de un día de edad. Las vacunas se administraron individualmente o en varias combinaciones y se evaluaron en relación con los controles no vacunados. Las evaluaciones se realizaron en los días 7 y 14 después de la vacunación (PV). Los criterios de valoración histológicos incluyeron puntuación semicuantitativa de la severidad de los componentes inflamatorios y determinaciones morfométricas cuantitativas de la concentración de células inflamatorias, el grosor de la mucosa y el porcentaje de superficie de la mucosa con cilios. Se observaron fuertes correlaciones positivas entre la puntuación rutinaria de severidad y los parámetros morfométricos inflamatorios, mientras que se observó una correlación negativa entre la severidad de la inflamación y el porcentaje de la superficie con cilios en la mucosa. Se observaron diferencias variables, a veces extensas y a menudo estadísticamente significativas en las respuestas inflamatorias entre las diversas vacunas. Una vacuna de la cepa de Massachusetts del virus de la bronquitis infecciosa (IBV-A) produjo la mayor respuesta inflamatoria general cuando se administró sola o en combinación con las vacunas de Newcastle. Se observó un aumento de la traqueítis en el día 14 después de la vacunación mediante la vacunación simultánea de la vacuna de bronquitis infecciosa A con las vacunas de Newcastle, pero no mediante la vacunación simultánea de la otra vacuna de la cepa Massachusetts (IBV-B) con Newcastle. Se observó una reducción en el porcentaje de los cilios para todos los grupos vacunados en comparación con los controles en el día siete después de la vacunación. Sin embargo, aunque la regeneración de cilios reactivos ocurrió en el día 14 después de la vacunación para la mayoría de los grupos vacunados, no fue evidente una respuesta de regeneración de cilios para la vacunación individual o combinada de Newcastle con la vacuna de bronquitis infecciosa Massachusetts A. El estudio también demuestra que puede estar presente una patología microscópica sustancial de la tráquea en aves vacunadas que no presentan signos respiratorios clínicos aparentes.

Genotypic and Phenotypic Factors Influencing Drug Response in Mexican Patients With Type 2 Diabetes Mellitus.

The treatment of Type 2 Diabetes Mellitus (T2DM) consists primarily of oral antidiabetic drugs (OADs) that stimulate insulin secretion, such as sulfonylureas (SUs) and reduce hepatic glucose production (e.g., biguanides), among others. The marked inter-individual differences among T2DM patients' response to these drugs have become an issue on prescribing and dosing efficiently. In this study, fourteen polymorphisms selected from Genome-wide association studies (GWAS) were screened in 495 T2DM Mexican patients previously treated with OADs to find the relationship between the presence of these polymorphisms and response to the OADs. Then, a novel association screening method, based on global probabilities, was used to globally characterize important relationships between the drug response to OADs and genetic and clinical parameters, including polymorphisms, patient information, and type of treatment. Two polymorphisms, ABCC8-Ala1369Ser and KCNJ11-Glu23Lys, showed a significant impact on response to SUs. Heterozygous ABCC8-Ala1369Ser variant (A/C) carriers exhibited a higher response to SUs compared to homozygous ABCC8-Ala1369Ser variant (A/A) carriers (p-value = 0.029) and to homozygous wild-type genotypes (C/C) (p-value = 0.012). The homozygous KCNJ11-Glu23Lys variant (C/C) and wild-type (T/T) genotypes had a lower response to SUs compared to heterozygous (C/T) carriers (p-value = 0.039). The screening of OADs response related genetic and clinical factors could help improve the prescribing and dosing of OADs for T2DM patients and thus contribute to the design of personalized treatments.

Comparison of real-time PCR with conventional PCR and culture to assess the efficacy of a live attenuated Salmonella enterica serovar Typhimurium vaccine against Salmonella enterica serovar Enteritidis in commercial leghorn chicks vaccinated under field and laboratory conditions.

The efficacy of a live attenuated Salmonella Typhimurium Megan Vac 1 vaccine (MV1) was evaluated against Salmonella Enteritidis in chicken pullets with the use of PCR and culture methods. Two hundred Hyline W-32 white leghorn chicks were obtained from a local hatchery and divided into four treatment groups. Two of the groups served as positive and negative controls. The MV1 vaccine was administered to the chicks in the remaining two groups at 1 and 35 days old by either the coarse spray (field) or the oral route (laboratory) method. The chicks were challenged with a high dose of a Salmonella Enteritidis strain at 10 wk old and euthanatized 3 days postinoculation. Samples for PCR analysis were collected prior to enrichment, after pre-enrichment in buffered peptone water (BPW) and after primary enrichment from the ceca, liver, and spleen. None of the samples tested yielded positive results for the Salmonella Typhimurium vaccine strain by either the culture or PCR methods. Results from the standard culture method showed that vaccinating the birds with MV1 reduced the counts of Salmonella Enteritidis recovered from the challenged birds. In addition, fewer pre-enriched samples tested positive for Salmonella Enteritidis among the challenged groups that were vaccinated when compared to the unvaccinated challenged group. Under the conditions of this study, MV1 was unable to prevent colonization of other internal organs such as the liver and spleen. Real-time PCR was significantly more sensitive than conventional PCR (C-PCR) prior to enrichment, but after enrichment the sensitivities of the two methods were similar. Enrichment significantly increased the sensitivity of both PCR methods for the detection of Salmonella Enteritidis in cecal samples, but did not significantly increase the sensitivity for detection of Salmonella Enteritidis in liver and spleen samples that were pre-enriched in BPW. There was no significant difference between the laboratory or field vaccination methods with respect to either the prevalence of Salmonella Enteritidis isolation or the bacterial loads in culture-positive samples. Collectively, the data suggest that MV1 offered some protection against Salmonella Enteritidis in commercial layer chick pullets under the conditions of this study. Given the labor and time required to perform the C-PCR and culture methods, the real-time PCR method may prove to be a more useful method to use in diagnostics.

A recombinant avian adeno-associated virus as a vector factor for infectious bursal disease vaccination

El virus de la enfermedad infecciosa de la bolsa es una enfermedad de distribución mundial que afecta a aves jóvenes que debe ser controlada mediante vacunación y constituye una de las preocupaciones principales de la industria avícola mundial. Los virus adeno-asociados aviares son virus no patogénicos capaces de dar cabida a porciones relativamente largas de ADN y de infectar una amplia variedad de tipos celulares. Un miembro de esta familia, el virus adeno-asociado aviar ha sido caracterizado por completo y utilizado como un vector para entrega de genes en células y tejidos de embriones de pollo. En el presente estudio se demostró mediante inmunohistoquímica y microscopia electrónica la factibilidad de generar virus adeno-asociados recombinantes expresando la proteína viral 2 del virus de la enfermedad infecciosa de la bolsa. Luego de la inoculación in ovo o intramuscular de aves libres de patógenos específicos con el virus recombinante, se observó evidencia serológica de la expresión de la proteína VP2. La utilización de virus adeno-asociado aviar recombinantes para la entrega de genes es una opción interesante para la vacunación de aves domésticas.

Infectious bursal disease is a worldwide distributed immunosuppressive disease of young chickens that need to be controlled by vaccination; it represents one of the main concerns for the poultry industry. The adeno-associated viruses are non-pathogenic viruses, capable of accommodating relatively long pieces of DNA, and of infecting a wide variety of cell types. A member of this family, the avian adeno-associated virus has been fully characterized and successfully used for gene delivery in chicken embryo tissues and cells. In this study, it was demonstrated by electron microscopy and immunocitochemistry the feasibility of generating recombinant avian adeno-associated virus (rAAAV) virions expressing the immunogenic viral protein 2 of infectious bursal disease virus (IBDV). Serological evidence of VP2 protein expression measured as IBDV specific antibody response after in ovo or intramuscular inoculation of the recombinant virus in specific pathogen free (SPF) chickens was observed. The use of rAAAV virions for gene delivery in poultry is a promising approach to poultry vaccination.

Molecular analysis of infectious bursal disease virus from bursal tissues collected on FTA filter paper.

We investigated the feasibility of using FTA filter cards for the storage of bursas of Fabricius containing infectious bursal disease virus (IBDV) and for IBDV detection by reverse transcriptase (RT)-polymerase chain reaction (PCR), and characterization by restriction fragment length polymorphism (RFLP) or nucleotide sequencing. The FTA card is a cotton-based cellulose membrane containing lyophilized chemicals that lyses many types of bacteria and viruses. IBDV was inactivated upon contact with the FTA as shown by the inability of the virus to be propagated in embryonating chicken eggs. Viral RNA in minced bursas or stamped bursas could be amplified by RT-PCR (VP2 gene fragment, 248 base pairs) after storage on FTA for at least 15 days at room temperature or 8 mo at -20 C. Analytical sensitivity of the test was between 0.5-5 ng of RNA template or 5 x 10(1) mean tissue culture infective dose (TCID50)/FTA spot. Detection rate of IBDV in domestic clinical samples collected on FTA or collected by the non-FTA standard procedure was 36.7% and 41.7%, respectively, which represents 88% agreement. Detection of IBDV from FTA cards inoculated with bursal tissues in the laboratory or in the field was 36.7% and 37.1%, respectively. Detection of IBDV from FTA samples when the cards were inoculated with bursal tissues and sent through customs into the United States was 32.9%. Analysis of the amplified products showed that molecular characterization of IBDV by RFLP or nucleotide sequencing is feasible in bursas stored on FTA at 25 C for 1-3 mo or at -20 C for at least 8 mo. The use of FTA for the collection of bursal tissues and simultaneous inactivation of IBDV allows the movement of specimens within the United States and also from outside the United States in compliance with federal regulations and in a manner adequate for molecular characterization.

Use of FTA filter paper for the molecular detection of Newcastle disease virus.

The feasibility of using Flinders Technology Associates filter papers (FTA cards) to collect allantoic fluid and chicken tissue samples for Newcastle disease virus (NDV) molecular detection was evaluated. Trizol RNA extraction and one-step reverse transcriptase-polymerase chain reaction (RT-PCR) were used. FTA cards allowed NDV identification from allantoic fluid with a titre of 10(5.8) median embryo lethal doses/ml. The inactivated virus remained stable on the cards for 15 days. NDV was detected from FTA imprints of the trachea, lung, caecal tonsil and cloacal faeces of experimentally infected birds. RT-PCR detection from FTA cards was confirmed by homologous frozen-tissue RT-PCR and virus isolation. Direct nucleotide sequence of the amplified F gene allowed prediction of NDV virulence. No virus isolation was possible from the FTA inactivated samples, indicating viral inactivation upon contact. The FTA cards are suitable for collecting and transporting NDV-positive samples, providing a reliable source of RNA for molecular characterization and a hazard-free sample.

Utilización de las tarjetas FTA® para el diagnóstico molecular del virus de la enfermedad de NEWCASTLE en muestras de fluido alantoideo / Utilization of FTA® Cards for the Molecular Diagnostic of Newcastle Disease Virus in Allantoic Fluid Samples

Se evaluó la factibilidad de utilizar las tarjetas FTA® (Flinders Technology Associates) para la inactivación y transporte de fluido alantoideo infectado con el virus de la enfermedad de Newcastle (VEN). La tarjetas FTA® fueron impregnadas con diluciones seriadas de fluido alantoideo con un título inicial de 10(8,8) DL/50 del VEN cepa LaSota y analizadas mediante la prueba de reacción en cadena por la polimerasa transcriptasa reversa (por sus siglas en ingles RT-PCR) a las 24 horas, 7 y 14 días. La concentración más baja del virus detectada en el fluido alantoideo fue 10(5,8) DL/50. La detección del virus a partir de la tarjeta fue posible hasta 14 días después de su inactivación. El re-aislamiento viral en huevos embrionados a partir de las tarjetas resultó negativo. La inactivación del virus en las tarjetas no afectó la calidad de la secuencia de nucleótidos, permitiendo la determinación de su virulencia mediante la secuenciación de los nucleótidos que codifican la zona de corte de la proteína de fusión resultando ser lentogénico en concordancia con la cepa inicial de virus aplicada a las tarjetas. Se concluye que las tarjetas FTA® representan una alternativa válida para el muestreo, inactivación y diagnóstico molecular del VEN, con un alto grado de bioseguridad.

The feasibility of using FTA® cards (Flinders Technology Associates) for inactivation and transportation of allantoic fluid infected with Newcastle disease virus (NDV) was evaluated. Serial dilutions of allantoic fluid with a titer of 10(8.8) ELD/50 of LaSota strain NDV were loaded on the FTA® cards and analyzed after 24 hour, 7 and 14 days using reverse transcriptase-polimerase chain reaction (RT-PCR). The lowest virus concentration that could be detected from the FTA® cards was 10(5.8) ELD/50. The detection of the inactivated virus was possible after 14 days of virus inactivation. No virus re-isolation in embryonating eggs was possible from the cards. No negative effects of the FTA® card inactivation on the nucleotide sequences were observed, allowing the determination of its virulence by direct nucleotide sequencing of the F protein cleavage site, resulting in a lentogénic strain in concordance with the initial virus applied on the cards. It can be conclude that FTA® cards are a valid alternative for NDV sampling, inactivation and molecular diagnostic with a high degree of biosecurity.