Imaging secondary neuronal degeneration.

OBJECTIVE: Distal nerve degeneration refers to the process of disintegration of a neuron or neuronal circuit as a consequence of distal damage. The end result of multiple etiologies, this finding is becoming common due to the increasing number of imaging tests done. This paper aims to define the different types of distal nerve damage, review the anatomy and function of the most commonly affected tracts, and illustrate distal nerve damage through diagrams and representative cases from routine practice. CONCLUSION: Knowing the distant response that can be expected according to the topography of a neuronal lesion is crucial to avoid diagnostic errors. Axonal degeneration and transsynaptic degeneration can be both antegrade and retrograde. Studies of cerebral metabolism, perfusion sequences, and diffusion sequences are showing increasingly earlier changes related to the same process; radiologists need to be aware of these changes.

Carretera perdida: imagen de la degeneración neuronal secundaria / Imaging secondary neuronal degeneration

Objetivo: La degeneración neuronal distal (DND) define el proceso de desintegración de una neurona o un circuito neuronal como consecuencia de una noxa localizada a distancia. Es un hallazgo frecuente debido al creciente número de realización de pruebas de imagen y a que refleja un desenlace común a múltiples etiologías. El objetivo de este artículo es definir los distintos tipos de daño neuronal a distancia, revisar la anatomía y función de los tractos más frecuentemente afectados e ilustrar la DND con esquemas y casos representativos de la práctica habitual. Conclusión: El conocimiento de la respuesta a distancia esperable según la topografía de una lesión neuronal es crucial para evitar errores diagnósticos. La degeneración axonal y transináptica puede ser tanto anterógrada como retrógrada. Los estudios de metabolismo cerebral, las secuencias de perfusión y de difusión muestran cambios cada vez más precoces del mismo proceso con los que debemos estar familiarizados.(AU

Objective: Distal nerve degeneration refers to the process of disintegration of a neuron or neuronal circuit as a consequence of distal damage. The end result of multiple etiologies, this finding is becoming common due to the increasing number of imaging tests done. This paper aims to define the different types of distal nerve damage, review the anatomy and function of the most commonly affected tracts, and illustrate distal nerve damage through diagrams and representative cases from routine practice. Conclusion: Knowing the distant response that can be expected according to the topography of a neuronal lesion is crucial to avoid diagnostic errors. Axonal degeneration and transsynaptic degeneration can be both antegrade and retrograde. Studies of cerebral metabolism, perfusion sequences, and diffusion sequences are showing increasingly earlier changes related to the same process; radiologists need to be aware of these changes.(AU)

LncRNAs in HCV Infection and HCV-Related Liver Disease.

Long non-coding RNAs (lncRNAs) are transcripts with poor coding capacity that may interact with proteins, DNA, or other RNAs to perform structural and regulatory functions. The lncRNA transcriptome changes significantly in most diseases, including cancer and viral infections. In this review, we summarize the functional implications of lncRNA-deregulation after infection with hepatitis C virus (HCV). HCV leads to chronic infection in many patients that may progress to liver cirrhosis and hepatocellular carcinoma (HCC). Most lncRNAs deregulated in infected cells that have been described function to potentiate or block the antiviral response and, therefore, they have a great impact on HCV viral replication. In addition, several lncRNAs upregulated by the infection contribute to viral release. Finally, many lncRNAs have been described as deregulated in HCV-related HCC that function to enhance cell survival, proliferation, and tumor progression by different mechanisms. Interestingly, some HCV-related HCC lncRNAs can be detected in bodily fluids, and there is great hope that they could be used as biomarkers to predict cancer initiation, progression, tumor burden, response to treatment, resistance to therapy, or tumor recurrence. Finally, there is high confidence that lncRNAs could also be used to improve the suboptimal long-term outcomes of current HCC treatment options.

[Long term evaluation of transanal surgery with automatic suture in Hirschsprung's disease]. / Evaluación a largo plazo de la cirugía transanal con sutura automática en la enfermedad de Hirschsprung.

OBJECTIVE: Retrospective and descriptive study of patients with diagnosis of Hirschsprung disease (HD) in whom transanal pull-through was performed in our center. MATERIAL AND METHODS: All patients with diagnosis of HD in whom transanal surgery was performed between 2006 and 2018 in our center were selected. In all cases laparoscopic biopsy was performed in the previous dilated area. Once intraoperatory biopsy revealed the presence of ganglionic cells, transanal De la Torre surgery was performed, until reaching the localization of the biopsied area. Circular automatic suture was performed in all cases. RESULTS: Surgery was performed in 21 patients (16 boys and 5 girls) with a median age of 12 months (5-62). No patient had enterocolitis. The median resection length was 14.5 cm (3-45) and no intraoperative complications happened. One patient (5%) developed a rectosigmoideal abscess, which was solved with antibiotic. One case (5%) needed another surgery due to umbilical trochar eventration. No patient had enterocolitis after the surgery. One patient (5%) had stenosis, solved by its section and other case (5%) had sleeve compression which was solved by laparoscopic section. After a median follow up of 97 months (12-159), one child (5%) developed encopresis, which is treated with periodic anal irrigations and other patient (5%) has chronic constipation, which require periodic enemas. The rest of the patients have no symptomatology. CONCLUSION: In conclusion, in our series of cases, transanal pull-through with circular automatic suture was a safe and useful technique. It has a low rate of operative and postoperative complications and provides good prospective results.

OBJETIVO: Estudio retrospectivo descriptivo de pacientes con enfermedad de Hirschsprung (EH) intervenidos en nuestro centro por vía transanal. MATERIAL Y METODOS: Se seleccionaron a los pacientes con diagnóstico de EH intervenidos por vía transanal con sutura automática circular en nuestro centro entre los años 2006 y 2018. Se realizó biopsia laparoscópica de la zona previa a la dilatación en todos los casos. Tras la confirmación de la presencia de células, se procedió al descenso transanal tipo De la Torre hasta alcanzar la zona de la biopsia. La anastomosis se realizó con sutura automática circular (CEEA 21 o 25 mm). RESULTADOS: Se intervinieron 21 pacientes, 16 niños y 5 niñas, con una mediana de edad de 12 meses (5-62). Ningún paciente presentó enterocolitis preoperatoria. La mediana de la longitud resecada fue de 14,5 cm (3-45), sin incidencias intraoperatorias. Un paciente (5%) presentó absceso rectosigmoideo postoperatorio, resuelto con antibióticos. Un caso (5%) necesitó de nueva cirugía por eventración del acceso umbilical. Un paciente (5%) presentó estenosis, resuelta mediante sección y otro (5%) compresión del manguito seromuscular, que fue tratada con sección laparoscópica. No hubo casos de enterocolitis postoperatoria. Tras una mediana de seguimiento de 97 meses (12-159), un niño presenta encopresis (5%) tratada mediante sistema de irrigaciones transanales periódicas y otro caso (5%) cursa con estreñimiento, en tratamiento con enemas periódicos. El resto se hallan asintomáticos. CONCLUSION: En definitiva, podemos afirmar que en nuestra serie el descenso transanal con sutura automática circular es una técnica segura y eficaz, que presenta una tasa baja de complicaciones intra y postoperatorias, con buenos resultados a largo plazo.

Evaluación a largo plazo de la cirugía transanal con sutura automática en la enfermedad de Hirschsprung / Long term evaluation of transanal surgery with automatic suture in Hirschsprung's disease

Objetivo. Estudio retrospectivo descriptivo de pacientes con en-fermedad de Hirschsprung (EH) intervenidos en nuestro centro por vía transanal. Material y métodos. Se seleccionaron a los pacientes con diagnóstico de EH intervenidos por vía transanal con sutura automática circular en nuestro centro entre los años 2006 y 2018. Se realizó biopsia laparoscópica de la zona previa a la dilatación en todos los casos. Tras la confirmación de la presencia de células, se procedió al descenso transanal tipo De la Torre hasta alcanzar la zona de la biopsia. La anastomosis se realizó con sutura automática circular (CEEA 21 o 25 mm). Resultados. Se intervinieron 21 pacientes, 16 niños y 5 niñas, con una mediana de edad de 12 meses (5-62). Ningún paciente presentó enterocolitis preoperatoria. La mediana de la longitud resecada fue de 14,5 cm (3-45), sin incidencias intraoperatorias. Un paciente (5%) presentó absceso rectosigmoideo postoperatorio, resuelto con antibióticos. Un caso (5%) necesitó de nueva cirugía por eventración del acceso umbilical. Un paciente (5%) presentó estenosis, resuelta mediante sección y otro (5%) compresión del manguito seromuscular, que fue tratada con sección laparoscópica. No hubo casos de enterocolitis postoperatoria. Tras una mediana de seguimiento de 97 meses (12-159), un niño presenta encopresis (5%) tratada mediante sistema de irrigaciones transanales periódicas y otro caso (5%) cursa con estreñimiento, en tratamiento con enemas periódicos. El resto se hallan asintomáticos. Conclusión. En definitiva, podemos afirmar que en nuestra serie el descenso transanal con sutura automática circular es una técnica segura y eficaz, que presenta una tasa baja de complicaciones intra y postoperatorias, con buenos resultados a largo plazo

Objective. Retrospective and descriptive study of patients with diagnosis of Hirschsprung disease (HD) in whom transanal pull-through was performed in our center. Material and methods. All patients with diagnosis of HD in whom transanal surgery was performed between 2006 and 2018 in our center were selected. In all cases laparoscopic biopsy was performed in the previous dilated area. Once intraoperatory biopsy revealed the presence of ganglionic cells, transanal De la Torre surgery was performed, until reaching the localization of the biopsied area. Circular automatic suture was performed in all cases. Results. Surgery was performed in 21 patients (16 boys and 5 girls) with a median age of 12 months (5-62). No patient had enterocolitis. The median resection length was 14.5 cm (3-45) and no intraoperative complications happened. One patient (5%) developed a rectosigmoideal abscess, which was solved with antibiotic. One case (5%) needed another surgery due to umbilical trochar eventration. No patient had enterocolitis after the surgery. One patient (5%) had stenosis, solved by its section and other case (5%) had sleeve compres-sion which was solved by laparoscopic section. After a median follow up of 97 months (12-159), one child (5%) developed encopresis, which is treated with periodic anal irrigations and other patient (5%) has chronic constipation, which require periodic enemas. The rest of the patients have no symptomatology. Conclusion. In conclusion, in our series of cases, transanal pull-through with circular automatic suture was a safe and useful technique. It has a low rate of operative and postoperative complications and provides good prospective results

¿Es posible el manejo ambulatorio de la invaginación intestinal? / Is it possible the outpatient management of intussusception?

Introducción: la invaginación intestinal es la causa más frecuente de obstrucción intestinal en menores de tres años. Habitualmente, tras la desinvaginación, los pacientes permanecen ingresados 24-48 horas. Recientemente se ha propuesto el manejo ambulatorio mediante observación clínica durante 12 horas. Nuestro objetivo es valorar la implementación de esta actitud terapéutica. Material y métodos: revisión retrospectiva de las invaginaciones intestinales atendidas en nuestro centro durante los últimos 12 años. Resultados: se incluye 458 pacientes, el 60,3% de ellos varones. Edad media de 24,1 meses, siendo la localización ileocólica la más frecuente (77,7%). El 2,4% presentó alguna causa secundaria. Se realizó neumoenema en 370 niños, requiriendo cirugía el 10,7%. Se registraron 78 recidivas en 56 pacientes (12,2%), 15 de ellos intrahospitalariamente. El tiempo medio para la reintroducción de la alimentación y la estancia media fueron de 28,6 y 64,4 horas respectivamente, sin diferencias significativas entre aquellos que recidivaron y los que no (60,8 frente a 69 horas; t = -0,4; p = 0,689). No se registraron diferencias entre el tiempo de evolución clínica y la tasa de éxito del neumoenema (t = 0,478; p = 0,634); aunque hubo diferencias en la necesidad de intervención quirúrgica (χ² = 5,604; p = 0,018), no hubo ninguna complicación. La reintroducción precoz de la alimentación no se relacionó con más recidivas ni diferencias entre los grupos (30,2% en el grupo que recidivó y 23,1% en el grupo sin recidiva, p = 0,608). Conclusiones: el ingreso hospitalario más allá de 12 horas no disminuye la tasa de complicaciones. Por tanto, consideramos que la observación en urgencias tras la desinvaginación durante 12 horas es una medida segura y coste-efectiva (AU)

Introduction: intussusception is the most frequent cause of bowel obstruction in children under three years. Usually, after reduction, patients remain admitted for 24-48 hours. Ambulatory management has recently been proposed, based on clinical experience of follow-up of the patient's evolution in the Emergency Department of the hospital during the following 12 hours. Our objective is to evaluate the implementation of this new therapeutic attitude. Material and methods: retrospective review of all the intussusceptions treated at our center during the last 12 years. Results: 458 patients were included, 60.3% ot them were male. Mean age was 24.1 months (SD 24.6), with the ileo-colic location being the most frequent (77.7%). 2.4% had secondary causes. A pneumoenema was performed in 370 children, requiring surgery 10.7%. There were 78 relapses in 56 patients (12.2%), 15 of them during admission. The mean time to reintroduce feeding and the mean hospital stay was 28.6 and 64.4 hours respectively, with no significant difference between those who relapsed and those who did not (60.8 vs 69 hours, t = -0.4, p = 0.689). There was no relationship between a longer clinical evolution and pneumoenema succeed rate (t = 0.478, p = 0.634). Although there were differences in the need for surgical intervention (χ² = 5.604, p = 0.018), there were no complications. Early reintroduction of feeding was not related to any recurrences or differences between groups (30.2% in the relapsed group and 23.1% in the non-recurrent group, p = 0.608). Conclusions: hospital admission beyond 12 hours does not decrease the rate of complications. Therefore, we consider that outpatient observation for 12 hours after reduction is a safe and economical measure (AU)

Adrenalectomía por vía laparoscópica: ventajas del abordaje mínimamente invasive / Laparoscopic adrenalectomy: advantages of the minimally invasive approach

Objetivo. Describir nuestra experiencia como centro terciario en el uso de la laparoscopia para la exéresis de lesiones adrenales en la edad pediátrica. Material y métodos. Estudio descriptivo y retrospectivo de pacientes con lesiones en la glándula suprarrenal intervenidos de suprarrenalectomía laparoscópica entre los años 2012 y 2015. Se estudiaron variables epidemiológicas, técnica quirúrgica, complicaciones y seguimiento. Resultados. Se estudiaron 9 pacientes con una mediana de 62 meses (5-184). Tres pacientes (33,33%) tenían un diagnóstico prenatal y tres pacientes debutaron con síntomas: hipertensión e insuficiencia renal, pubertad precoz y otro con un adenoflemón secundario a metástasis. En el resto, el hallazgo fue incidental. En cinco pacientes la lesión se situó en el lado derecho y en cuatro en el izquierdo. La intervención se realizó por vía laparoscópica. En un paciente se convirtió a cirugía abierta por mala visualización en el campo quirúrgico. No se produjeron complicaciones perioperatorias ni necesidad de trasfusión. La media de la estancia hospitalaria fue de 4 días (2-5). El dolor postoperatorio fue controlado durante las primeras 24 horas y con analgésicos de primer escalón. Los diagnósticos confirmados por histología fueron: seis neuroblastomas, dos ganglioneuromas y un adenoma. La media de seguimiento fue de 22 meses (1-53). Conclusiones. En nuestra serie esta vía de abordaje está asociada a una baja morbimortalidad con una recuperación precoz, corta estancia hospitalaria y resultados satisfactorios. Por tanto, proponemos la adrenalectomía laparoscópica como una buena alternativa para el abordaje de las lesiones suprarrenales en la población pediátrica

Objective. To describe our experience as a tertiary center on the use of laparoscopic adrenal surgery in children. Material and Methods. A descriptive, retrospective study of patients with pathologic adrenal masses undergoing laparoscopic adrenal surgery, between 2012 and 2015. Epidemiological variables, surgical technique, complications and follow-up were studied. Results. Nine patients were studied with a median age of 62 months (5-184). In 3 patients (33.33%) there was a prenatal diagnosis. Three patients had symptoms: hypertension and renal failure, precocious puberty, and an infected lymph node secondary to metastasis. In the rest, findings were incidental. In five patients the lesion was located on the right side and in four, on the left. The operation was performed laparoscopically. There was one conversion to open surgery because of poor visualization of the surgical field and no major intra- or postoperative complications were noted. The average hospital stay was 4 days (2-5). Postoperative pain was controlled during the first 24 hours with first step painkillers. Diagnoses were histologically confirmed: two ganglioneuroma, three neuroblastoma, bronchogenic cyst, pulmonary sequestration and adenoma. Mean follow-up was 22 months (1-53). Conclusions. In our series this surgical approach is associated with low morbidity and mortality, early recovery, shorter hospital stay and satisfactory results. We therefore propose laparoscopic adrenalectomy as a good alternative for approaching the pathologic adrenal masses in the pediatric population

[Laparoscopic adrenalectomy: advantages of the minimally invasive approach]. / Adrenalectomía por vía laparoscópica: ventajas del abordaje mínimamente invasivo.

AIM: To describe our experience as a tertiary center on the use of laparoscopic adrenal surgery in children. MATERIAL AND METHODS: A descriptive, retrospective study of patients with pathologic adrenal masses undergoing laparoscopic adrenal surgery, between 2012 and 2015. Epidemiological variables, surgical technique, complications and follow-up were studied. RESULTS: Nine patients were studied with a median age of 62 months (5-184). In 3 patients (33.33%) there was a prenatal diagnosis. Three patients had symptoms: hypertension and renal failure, precocious puberty, and an infected lymph node secondary to metastasis. In the rest, findings were incidental. In five patients the lesion was located on the right side and in four, on the left. The operation was performed laparoscopically. There was one conversion to open surgery because of poor visualization of the surgical field and no major intra- or postoperative complications were noted. The average hospital stay was 4 days (2-5). Postoperative pain was controlled during the first 24 hours with first step painkillers. Diagnoses were histologically confirmed: two ganglioneuroma, three neuroblastoma, bronchogenic cyst, pulmonary sequestration and adenoma. Mean follow-up was 22 months (1-53). CONCLUSIONS: In our series this surgical approach is associated with low morbidity and mortality, early recovery, shorter hospital stay and satisfactory results. We therefore propose laparoscopic adrenalectomy as a good alternative for approaching the pathologic adrenal masses in the pediatric population.

OBJETIVO: Describir nuestra experiencia como centro terciario en el uso de la laparoscopia para la exéresis de lesiones adrenales en la edad pediátrica. MATERIAL Y METODOS: Estudio descriptivo y retrospectivo de pacientes con lesiones en la glándula suprarrenal intervenidos de suprarrenalectomía laparoscópica entre los años 2012 y 2015. Se estudiaron variables epidemiológicas, técnica quirúrgica, complicaciones y seguimiento. RESULTADOS: Se estudiaron 9 pacientes con una mediana de 62 meses (5-184). Tres pacientes (33,33%) tenían un diagnóstico prenatal y tres pacientes debutaron con síntomas: hipertensión e insuficiencia renal, pubertad precoz y otro con un adenoflemón secundario a metástasis. En el resto, el hallazgo fue incidental. En cinco pacientes la lesión se situó en el lado derecho y en cuatro en el izquierdo. La intervención se realizó por vía laparoscópica. En un paciente se convirtió a cirugía abierta por mala visualización en el campo quirúrgico. No se produjeron complicaciones perioperatorias ni necesidad de trasfusión. La media de la estancia hospitalaria fue de 4 días (2-5). El dolor postoperatorio fue controlado durante las primeras 24 horas y con analgésicos de primer escalón. Los diagnósticos confirmados por histología fueron: seis neuroblastomas, dos ganglioneuromas y un adenoma. La media de seguimiento fue de 22 meses (1-53). CONCLUSIONES: En nuestra serie esta vía de abordaje está asociada a una baja morbimortalidad con una recuperación precoz, corta estancia hospitalaria y resultados satisfactorios. Por tanto, proponemos la adrenalectomía laparoscópica como una buena alternativa para el abordaje de las lesiones suprarrenales en la población pediátrica.

iHWG-µNIR: a miniaturised near-infrared gas sensor based on substrate-integrated hollow waveguides coupled to a micro-NIR-spectrophotometer.

A miniaturised gas analyser is described and evaluated based on the use of a substrate-integrated hollow waveguide (iHWG) coupled to a microsized near-infrared spectrophotometer comprising a linear variable filter and an array of InGaAs detectors. This gas sensing system was applied to analyse surrogate samples of natural fuel gas containing methane, ethane, propane and butane, quantified by using multivariate regression models based on partial least square (PLS) algorithms and Savitzky-Golay 1(st) derivative data preprocessing. The external validation of the obtained models reveals root mean square errors of prediction of 0.37, 0.36, 0.67 and 0.37% (v/v), for methane, ethane, propane and butane, respectively. The developed sensing system provides particularly rapid response times upon composition changes of the gaseous sample (approximately 2 s) due the minute volume of the iHWG-based measurement cell. The sensing system developed in this study is fully portable with a hand-held sized analyser footprint, and thus ideally suited for field analysis. Last but not least, the obtained results corroborate the potential of NIR-iHWG analysers for monitoring the quality of natural gas and petrochemical gaseous products.

U1 snRNP control of 3'-end processing and the therapeutic application of U1 inhibition combined with RNA interference.

U1 snRNP (U1 small nuclear ribonucleoprotein) is a well-characterized splicing factor. Besides, when U1 snRNP binds close to a putative polyadenylation site, mostly located in introns, it prevents premature cleavage and polyadenylation and controls the length of most cellular mRNAs. On the other hand, U1 snRNP binding close to the 3'-end of some mRNAs, inhibits polyadenylation and, therefore, gene expression. The inhibition of polyadenylation by U1 snRNP is the basis of U1i (U1 snRNP-based inhibition), a technique used to inhibit gene expression. U1i consists of the expression of a U1 snRNP modified to interact with a target mRNA and inhibit target gene expression. U1i has been used to inhibit the expression of reporter or endogenous genes both in tissue culture and in animal models. Furthermore, combination of U1i and RNA interference (RNAi) results in synergistic increased inhibitions which allow the dose of inhibitors to be decreased whilst at the same time obtaining good inhibitions with fewer unwanted secondary effects. The combination of RNAi and U1i is of special interest for antiviral therapy, as a functional decrease of the expression of replicative viral RNAs may require high inhibition and the combination of two or more inhibitors should decrease the possibility of escape mutants resistant to treatment. In fact, a therapy with U1i combined with RNAi is currently being developed for the treatment of HBV infections. We believe that this review will clarify the hallmarks of U1i technology and will encourage many laboratories to use U1i for functional studies and therapeutic applications.

Tercer reporte de eventos adversos con tratamientos biológicos en Argentina. Informe de registro biobadasar / Thrid report of adverse events with biologics in Argentina. Report from biobadasar registry

Introducción: BIOBADASAR (Registro Argentino de Eventos Adversos con Tratamientos Biológicos en Reumatología) comenzó en agosto de 2010. La importancia de este registro es mostrar datos locales que, probablemente, puedan diferir de otros registros. El objetivo es comunicar los resultados del tercer reporte de BIOBADASAR. Métodos: Todos los pacientes con enfermedades reumáticas que requirieron tratamiento con agentes biológicos y pacientes controles sin estos tratamientos fueron incluidos en la base de datos provenientes de 32 centros participando a lo largo de la Argentina. Tres áreas de datos son analizados: características de los pacientes, tratamientos y eventos adversos...

Introduction: BIOBADASAR (Argentine Registry of Adverse Events with Biological Treatments in Rheumatology) began in August 2010. The importance of this registry is to show local data that may probably differ from other registries. The objective is to communicate the results of the third BIOBADASAR report. Methods: All patients with rheumatic diseases who required treatment with biological agents and control patients without these treatments were included in the database from 32 participating centers throughout Argentina. Three areas of data are analyzed: patient characteristics, treatments and adverse events...

AAV vectors transduce hepatocytes in vivo as efficiently in cirrhotic as in healthy rat livers.

In liver cirrhosis, abnormal liver architecture impairs efficient transduction of hepatocytes with large viral vectors such as adenoviruses. Here we evaluated the ability of adeno-associated virus (AAV) vectors, small viral vectors, to transduce normal and cirrhotic rat livers. Using AAV serotype-1 (AAV1) encoding luciferase (AAV1Luc) we analyzed luciferase expression with a CCD camera. AAV1Luc was injected through the hepatic artery (intra-arterial (IA)), the portal vein (intra-portal (IP)), directly into the liver (intra-hepatic (IH)) or infused into the biliary tree (intra-biliar). We found that AAV1Luc allows long-term and constant luciferase expression in rat livers. Interestingly, IP administration leads to higher expression levels in healthy than in cirrhotic livers, whereas the opposite occurs when using IA injection. IH administration leads to similar transgene expression in cirrhotic and healthy rats, whereas intra-biliar infusion is the least effective route. After 70% partial hepatectomy, luciferase expression decreased in the regenerating liver, suggesting lack of efficient integration of AAV1 DNA into the host genome. AAV1Luc transduced mainly the liver but also the testes and spleen. Within the liver, transgene expression was found mainly in hepatocytes. Using a liver-specific promoter, transgene expression was detected in hepatocytes but not in other organs. Our results indicate that AAVs are convenient vectors for the treatment of liver cirrhosis.

AAV-mediated in vivo knockdown of luciferase using combinatorial RNAi and U1i.

RNA interference (RNAi) has been successfully employed for specific inhibition of gene expression; however, safety and delivery of RNAi remain critical issues. We investigated the combinatorial use of RNAi and U1 interference (U1i). U1i is a gene-silencing technique that acts on the pre-mRNA by preventing polyadenylation. RNAi and U1i have distinct mechanisms of action in different cellular compartments and their combined effect allows usage of minimal doses, thereby avoiding toxicity while retaining high target inhibition. As a proof of concept, we investigated knockdown of the firefly luciferase reporter gene by combinatorial use of RNAi and U1i, and evaluated their inhibitory potential both in vitro and in vivo. Co-transfection of RNAi and U1i constructs showed additive reduction of luciferase expression up to 95% in vitro. We attained similar knockdown when RNAi and U1i constructs were hydrodynamically transfected into murine liver, demonstrating for the first time successful in vivo application of U1i. Moreover, we demonstrated long-term gene silencing by AAV-mediated transduction of murine muscle with RNAi/U1i constructs targeting firefly luciferase. In conclusion, these results provide a proof of principle for the combinatorial use of RNAi and U1i to enhance target gene knockdown in vivo.

Ovary development and maturation in Nezara viridula (L.) (Hemiptera: Pentatomidae).

Ovary development and maturation of Nezara viridula (L.) were evaluated by examining ovariole morphology and the alterations in the biochemical (protein synthesis related to reproduction) composition of the hemolymph. Quantitative and qualitative protein analyses were performed and ovary structural alterations for the pre-reproductive and reproductive stages were recorded. Total concentration of proteins in female hemolymph gradually increased until the end of the pre-mating stage, remaining unaltered thereafter. Proteins linked to reproduction (vitellogenins) appeared in the hemolymph 10 days after adult emergence and indicated the end of the pre-mating stage. After mating, total protein concentration in the hemolymph was lower compared to virgin females; vitellogenin levels were similar during most of the observation period. Oocyte development and maturation were gradual and age dependent. Ten-day-old females had chorionated oocytes ready for fertilization. Mating did not stimulate oocyte development in N. viridula, but the lack of mating activity appeared to have stimulated oocyte resorption in 17-day-old females.

Ovary Development and Maturation in Nezara viridula (L.) (Hemiptera: Pentatomidae)

Ovary development and maturation of Nezara viridula (L.) were evaluated by examining ovariole morphology and the alterations in the biochemical (protein synthesis related to reproduction) composition of the hemolymph. Quantitative and qualitative protein analyses were performed and ovary structural alterations for the pre-reproductive and reproductive stages were recorded. Total concentration of proteins in female hemolymph gradually increased until the end of the pre-mating stage, remaining unaltered thereafter. Proteins linked to reproduction (vitellogenins) appeared in the hemolymph 10 days after adult emergence and indicated the end of the pre-mating stage. After mating, total protein concentration in the hemolymph was lower compared to virgin females; vitellogenin levels were similar during most of the observation period. Oocyte development and maturation were gradual and age dependent. Ten-day-old females had chorionated oocytes ready for fertilization. Mating did not stimulate oocyte development in N. viridula, but the lack of mating activity appeared to have stimulated oocyte resorption in 17-day-old females.

Combination of RNA interference and U1 inhibition leads to increased inhibition of gene expression.

RNA interference (RNAi) has been revolutionary for the specific inhibition of gene expression. However, the application of RNAi has been hampered by the fact that many siRNAs induce dose-dependent unwanted secondary effects. Therefore, new methods to increase inhibition of gene expression with low doses of inhibitors are required. We have tested the combination of RNAi and U1i (U1 small nuclear RNA--snRNA--interference). U1i is based on U1 inhibitors (U1in), U1 snRNA molecules modified to target a pre-mRNA and inhibit its gene expression by blocking nuclear polyadenylation. The combination of RNAi and U1i resulted in stronger inhibition of reporter or endogenous genes than that obtained using either of the techniques alone. The increased inhibition observed is stable over time and allows higher inhibition than the best obtained with either of the inhibitors alone even with decreased doses of the inhibitors. We believe that the combination of RNAi and U1i will be of interest when higher inhibition is required or when potent inhibitors are not available. Also, the combination of these techniques would allow functional inhibition with a decreased dose of inhibitors, avoiding toxicity due to dose-dependent unwanted effects.

Liver transduction with a simian virus 40 vector encoding insulin-like growth factor I reduces hepatic damage and the development of liver cirrhosis.

Liver transplantation is the only treatment for advanced liver cirrhosis. Therapies halting the progression of the disease are urgently needed. Administration of recombinant insulin-like growth factor-I (rIGF-I) induces hepatoprotective effects in experimental cirrhosis. Therefore, we analyzed the efficacy of a recombinant simian virus 40 vector (rSV40) encoding IGF-I (rSVIGF-I) to prevent cirrhosis progression. First, transgene expression was evaluated in mice injected with rSV40 encoding luciferase, which showed long-term hepatic expression of the transgene. Interestingly, luciferase expression increased significantly in CCl(4)-damaged livers and upon IGF-I administration, thus liver injury and IGF-I expression from rSVIGF-I should favor transgene expression. rSVIGF-I therapeutic efficacy was studied in rats where liver cirrhosis was induced by CCl(4) inhalation during 36 weeks. At the end of the study, the hepatic levels of IGF-I and IGF-binding protein 3 were higher in rSVIGF-I-treated rats than in control cirrhotic animals. Cirrhotic rats treated with rSVIGF-I had reduced serum bilirubin, transaminases and liver fibrosis scores and increased hepatic expression of hepatocyte growth factor and STAT3alpha as compared to cirrhotic animals. Furthermore, cirrhotic animals showed testis atrophy and altered spermatogenesis, whereas testicular size and histology were normal in cirrhotic rats that received rSVIGF-I. Therefore, rSV40-mediated sustained expression of IGF-I in the liver slowed cirrhosis progression.

Platynota rostrana (Walker) (Tortricidae) and Phidotricha erigens Raganot (Pyralidae): artificial diet effects on biological cycle

The lepidopterans Platynota rostrana (Walker) (Tortricidae) and Phidotricha erigens Raganot (Pyralidae) have been found frequently in citrus groves in São Paulo State in recent years. Since in Brazil, the fertility cycle of these two species is largely unknown, as are details of the damage wrought by them in crops, this research studied these aspects of the two species, which were kept under laboratory conditions (temperature 25 ± 2 °C, 70 ± 10 percent RH, 14 h photophase) and on an artificial diet. The duration of the biological cycle (egg-adult) for P. rostrana was 38.3 days and total viability was 44.0 percent; for P. erigens these values were 32.5 days and 63.6 percent, respectively. Both species showed five larval instars. Females of P. rostrana laid an average of 308 eggs, whereas those of P. erigens laid an average of 106 eggs. In both species, female pupae were heavier than males. Male and female longevity for both species was nearly 10 days. Based on the data obtained, the artificial diet produced better results in P. rostrana than in P. erigens. If these species, which have the potential to reach pest status in the citrus groves of São Paulo State, could be reared on an artificial diet, research on their control by alternative methods would be easier.

Os lepidópteros Platynota rostrana (Walker) (Tortricidae) e Phidotricha erigens Raganot (Pyralidae) têm sido constatados com freqüência nos pomares cítricos do Estado de São Paulo, nos últimos anos. O objetivo deste trabalho foi estudar a biologia das duas espécies, em condições de laboratório (temperatura 25 ± 2 °C, UR. 70 ± 10 por cento e fotofase de 14 h) em dieta artificial, elaborar uma tabela de vida de fertilidade e descrever os danos causados no campo, devido ao desconhecimento destes aspectos biológicos dos referidos insetos no Brasil. A duração do ciclo biológico (ovo-adulto) de P. rostrana foi de 38,3 dias e a viabilidade total de 44,0 por cento, enquanto para P. erigens foi de 32,5 dias e 63,6 por cento, respectivamente. Ambas as espécies apresentaram cinco ínstares e as pupas de fêmeas foram mais pesadas do que as de machos. As fêmeas de P. rostrana colocaram, em média, 308 ovos e as de P. erigens 106 ovos. A longevidade de machos e fêmeas das duas espécies foi próxima de 10 dias. Pela tabela de vida de fertilidade concluiu-se que P. rostrana tem melhor desempenho em dieta artificial que P. erigens. É possível criar estas espécies em dieta artificial, facilitando o desenvolvimento de pesquisas relacionadas ao seu controle por métodos alternativos, caso elas assumam o status de pragas nos pomares de São Paulo.

Platynota rostrana (Walker) (Tortricidae) AND Phidotricha erigens Raganot (Pyralidae): artificial diet effects on biological cycle.

The lepidopterans Platynota rostrana (Walker) (Tortricidae) and Phidotricha erigens Raganot (Pyralidae) have been found frequently in citrus groves in São Paulo State in recent years. Since in Brazil, the fertility cycle of these two species is largely unknown, as are details of the damage wrought by them in crops, this research studied these aspects of the two species, which were kept under laboratory conditions (temperature 25 +/- 2 degrees C, 70 +/- 10% RH, 14 h photophase) and on an artificial diet. The duration of the biological cycle (egg-adult) for P. rostrana was 38.3 days and total viability was 44.0%; for P. erigens these values were 32.5 days and 63.6%, respectively. Both species showed five larval instars. Females of P. rostrana laid an average of 308 eggs, whereas those of P. erigens laid an average of 106 eggs. In both species, female pupae were heavier than males. Male and female longevity for both species was nearly 10 days. Based on the data obtained, the artificial diet produced better results in P. rostrana than in P. erigens. If these species, which have the potential to reach pest status in the citrus groves of São Paulo State, could be reared on an artificial diet, research on their control by alternative methods would be easier.

Comparison of high pressure-induced dissociation of single-stranded DNA-binding protein (SSB) from high pressure-sensitive and high pressure-adapted marine Shewanella species.

The effects of hydrostatic pressure on protein quaternary structure were compared for recombinant single-stranded DNA-binding protein (SSB) derived from piezosensitive, piezotolerant, and obligately piezophilic ("pressure-loving") marine Shewanella strains. The pressure-induced dissociation of the oligomeric SSB proteins was investigated using fluorescence anisotropy. The SSBs all exhibited striking similarity in the pressure-dependent behavior of the fluorescence intensity and emission spectrum as well as in their dissociation constants at atmospheric pressure. The free energies of subunit association into tetramers for all SSBs were between -27 and -30 kcal mol(-1). However, SSB from the piezosensitive Shewanella strain S. hanedai was more sensitive to pressure than that of the SSB proteins from the piezotolerant or piezophilic bacteria. The volume change of association obtained from the pressure dependence of dissociation at a fixed protein concentration (Delta V(p)) for SSB from S. hanedai was 394-402 ml mol(-1). The Delta V(p) values for SSB from the deeper-living Shewanellas were smaller and ranged from 253 to 307 ml mol(-1). Differences between the primary structures of the SSB proteins that could correlate with differences in sensitivity to pressure-induced dissociation were examined.