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Polylactic Acid (PLA) and Acrylonitrile-Butadiene-Styrene (ABS) are commonly used polymers in 3D printing for biomedical applications. Dental Pulp Stem Cells (DPSCs) are an accessible and proliferative source of stem cells with significant differentiation potential. Limited knowledge exists regarding the biocompatibility and genetic safety of ABS and PLA when in contact with DPSCs. This study aimed to investigate the impact of PLA and ABS on the adhesion, proliferation, osteogenic differentiation, genetic stability, proteomics, and immunophenotypic profile of DPSCs. A total of three groups, 1- DPSC-control, 2- DPSC+ABS, and 3- DPSC+PLA, were used in in vitro experiments to evaluate cell morphology, proliferation, differentiation capabilities, genetic stability, proteomics (secretome), and immunophenotypic profiles regarding the interaction between DPSCs and polymers. Both ABS and PLA supported the adhesion and proliferation of DPSCs without exhibiting significant cytotoxic effects and maintaining the capacity for osteogenic differentiation. Genetic stability, proteomics, and immunophenotypic profiles were unaltered in DPSCs post-contact with these polymers, highlighting their biosafety. Our findings suggest that ABS and PLA are biocompatible with DPSCs and demonstrate potential in dental or orthopedic applications; the choice of the polymer will depend on the properties required in treatment. These promising results stimulate further studies to explore the potential therapeutic applications in vivo using prototyped polymers in personalized medicine.

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Traumatic spinal cord injury (SCI) is a devastating condition without an effective therapy. Cellular therapies are among the promising treatment strategies. Adult stem cells, such as mesenchymal stem cells, are often used clinical research for their immunomodulatory and regenerative potential. This study aimed to evaluate the effect of human adipose tissue-derived stem cells (ADSC) infusion through the cauda equina in rats with SCI. The human ADSC from bariatric surgery was isolated, expanded, and characterized. Wistar rats were subjected to blunt SCI and were divided into four groups. Two experimental groups (EG): EG1 received one ADSC infusion after SCI, and EG2 received two infusions, the first one after SCI and the second infusion seven days after the injury. Control groups (CG1 and CG2) received infusion with a culture medium. In vivo, cell tracking was performed 48 h and seven days after ADSC infusion. The animals were followed up for 40 days after SCI, and immunohistochemical quantification of myelin, neurons, and astrocytes was performed. Cellular tracking showed cell migration towards the injury site. ADSC infusion significantly reduced neuronal loss, although it did not prevent the myelin loss or enhance the area occupied by astrocytes compared to the control group. The results were similar when comparing one or two cell infusions. The injection of ADSC distal to the injured area was shown to be a safe and effective method for cellular administration in spinal cord injury.

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Abstract: Asthma is a chronic respiratory disease affecting 300 million people worldwide. It results in several structural changes in the airways, which are minimally accessible in clinical practice. Cell therapy using mesenchymal stromal cells (MSCs) is a promising strategy for treating asthma due to the paracrine activity of MSCs, which influences tissue regeneration and modulates the immune response. Studies using extracellular vesicles (EV) released by MSCs have demonstrated their regenerative properties in animal models. The aim of this study was to evaluate the potential of EVs isolated from human bone marrow MSCs (hBM-MSCs) to control lung tissue remodeling in ovalbumin-induced allergic asthma in Balb/c mice. We isolated hBM-MSCs from a single donor, expanded and characterized them, and then isolated EVs. Asthma was induced in 43 male Balb/c mice, divided into four groups: control, asthmatic (AS), asthmatic plus systemic EVs (EV-S), and asthmatic plus intratracheal EVs (EV-IT). Upon completion of asthma induction, animals were treated with EVs either locally (EV-IT) or intravenously (EV-S). Seven days after, we performed bronchoalveolar lavage (BAL) and the total nuclear cells were counted. The animals were euthanized, and the lungs were collected for histopathological analysis of the airways. The EV-S group showed improvement in only the total BAL cell count compared with the AS group, while the EV-IT group showed significant improvement in almost all evaluated criteria. Therefore, we demonstrate that the local application of EVs derived from hBM-MSCs may be a potential treatment in controlling asthma.

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Las mujeres con trasplante renal (TR) suelen cursar un embarazo de alto riesgo, pues son más susceptibles a presentar complicaciones médicas. Aunado a este hecho pueden presentar alteraciones en su estado emocional y psicológico durante y después del embarazo, lo que impactaría directamente en el estilo de crianza y desarrollo posterior del niño. Se desconoce cómo son los estilos de crianza en los hijos de las mujeres con TR y el posible impacto en su funcionamiento cognoscitivo. El presente trabajo tuvo dos objetivos: 1) determinar si existían diferencias en los estilos de crianza y las funciones cognoscitivas de los hijos de mujeres con trasplante renal en comparación con los hijos de mujeres sanas y 2) identificar la asociación entre las variables asociadas al riesgo en el embarazo y los estilos de crianza con las funciones cognoscitivas. Para ello fueron reclutados de diversos centros de trasplantes 23 niños (9 casos y 14 controles) de 7-15 años con sus respectivas madres a quienes se evaluó mediante el WISC-IV y el cuestionario CRPBI. Las madres fueron evaluadas con el cuestionario BRIEF, el cuestionario PCRI-M y el BDI-II. Se realizó un diseño transversal con alcance correlacional. No se encontraron diferencias entre el grupo caso y control en los estilos de crianza ni en las funciones cognoscitivas. Sin embargo, sí se encontró asociación entre los estilos de crianza y las funciones cognoscitivas. Un estilo en el que predomine la comunicación y la disciplina se asocia con mejores resultados cognoscitivos y conductuales.

Pregnancy in Kidney Transplant (KT) recipients is usually considered of high risk. KT recipients are susceptible to have obstetric complications. Studies report that KT recipients are at higher risk of experiencing distress during and after pregnancy, situation that may have an impact in parenting styles and child's development. Characteristics of parenting styles in KT recipients' offspring and its possible impact in cognitive functions remain unknown. The aims of the study were: 1) to determine if there are differences in parenting styles and cognitive functions among KT recipients' offspring and healthy women's offspring, and 2) to identify associations between high risk pregnancy variables, parenting styles and cognitive functions. Twenty-three children (9 cases and 14 controls) aged 8-15 years and their mothers were assessed with the (WISC-V) and the CRPBI. Mothers were asked to complete BRIEF, PCRI-M, BDI-II and a socioeconomic status instrument. A transversal correlational design was performed. No differences were found between case and control group in parenting styles or cognitive functions. However, an association between parenting styles and cognitive functions was found. Being raised with communication and discipline is associated with increased cognitive results.

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La enfermedad de Kawasaki (EK) es una vasculitis aguda de vaso mediano que afecta principalmente a niños menores de 5 años, que de no ser tratada, se asocia al desarrollo de aneurismas de las arterias coronarias en aproximadamente el 25% de los casos. Típicamente la EK se presenta con fiebre, cambios mucocutáneos y linfadenopatía. Sin embargo, EK es una enfermedad excepcional en la que las formas incompletas de la enfermedad son muy comunes y a menudo asociación sintomatologías atípicas. Éstas pueden crear un desafío diagnóstico para los tratantes y retrasar el inicio de la terapia. No existe un gold standard para el diagnóstico de EK pero hay estudios de laboratorio y hallazgos ecocardiográficos que permiten apoyar el diagnóstico en casos incompletos. El estudio con ecocardiograma debe realizarse lo más pronto posible cuando se sospecha el diagnóstico, pero no debe retrasar el inicio de tratamiento. El objetivo de la terapia en EK es disminuir el riesgo de desarrollar aneurismas de las arterias coronarias y de esta forma, la morbimortalidad asociada a dicha condición. El propósito de esta revisión es conocer las características clínicas y las posibles formas de presentación de esta patología, además del tratamiento actual.

Kawasaki disease (KD) is an acute vasculitis of the medium vessel that mainly affects children under 5 years old, which if it's not treated, is associated with the development of coronary artery aneurysms in approximately 25% of all cases. Typically, KD presents with fever, mucocutaneous changes and lymphadenopathy. However, EK is an exceptional disease in which incomplete forms of the disease are very common and atypical presentations often occur. These presentations may create a diagnostic puzzle for pediatricians and may delay the start of therapy. There is no specific study for the diagnosis of KD but there are laboratory studies and findings in ECG that support the diagnosis in atypical cases. The echocardiogram study should be performed as soon as possible when the diagnosis is suspected, but should not delay the treatment. The goal of treatment in KD is to reduce the risk of developing aneurysms of the coronary arteries and thereby decrease the morbidity and mortality associated with this condition.

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La parotiditis es un infección viral producida por el virus parotídeo. Clínicamente se caracteriza por aumento de volumen de la glándula parótida generalmente bilateral. La estrategia que ha mostrado ser más eficaz para la prevención de esta infección ha sido la implementación de la vacuna tres vírica en los programas de inmunización. En países con población altamente inmunizada como Chile, se logró una importante disminución de la incidencia de esta enfermedad. Sin embargo, a pesar de la efectividad de la vacuna se siguen reportando brotes en todo el mundo, evidenciándose un cambio epidemiológico, trasladándose la edad de presentación clínica desde la niñez y adolescencia hacia los adultos jóvenes. Este aumento en el número de casos ha sido estudiado, determinando que el efecto protector inmunitario de la vacuna decaería con el transcurso del tiempo, contribuyendo a la propagación de los brotes. Con respecto a posibles estrategias para el manejo de los brotes la aplicación de una dosis adicional de la vacunas tres vírica en población expuesta sería una medida que mejoraría el control de los brotes.

Mumps is a viral infection caused by mumps virus. Clinically, it is characterized by increased parotid volume. The most effective strategy for preventing this infection, has been the implementation of measles-mumps-rubella (MMR) vaccine in the national immunization program. Among countries with a highly immunized population, like Chile, there has been an important reduction in the incidence of this disease. Nevertheless, despite the effectivity of the MMR, there are reports of outbreaks worldwide, with an epidemiological change, from clinical presentation in childhood, to adolescents and adults. This outbreaks have been studied, and it has been determined that they are due to the waning of vaccine-derived immunity. Regarding strategies for the management of new outbreaks, the administration of an additional dose of MMR, would be an alternative.

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Perfluorooctanoic acid (PFOA) is a member of the perfluoroalkyl acid family of compounds. Due to the presence of strong carbon-fluorine bonds, it is practically nonbiodegradable and highly persistent in the environment. PFOA has been detected in the follicular fluid of women, and positively associated with reduced fecundability and infertility. However, there are no reports concerning the experimental evaluation of PFOA on oocyte toxicity in mammals. The aim of the present study was to determine if PFOA is able to induce oxidative stress in fetal ovaries and cause apoptosis in oocytes in vitro. In addition, since inhibition of the gap junction intercellular communication (GJIC) by PFOA has been demonstrated in liver cells in vivo and in vitro, the effect of PFOA on the GJIC between the oocyte and its supportive cumulus cells was studied. Results show that PFOA induced oocyte apoptosis and necrosis in vitro (medium lethal concentration, LC50 = 112.8 µM), as evaluated with Annexin-V-Alexa 508 in combination with BOBO-1 staining. Reactive oxygen species (ROS) levels, as assessed by DCFH-DA, increased significantly in fetal ovaries exposed to » LC50 (28.2 µM, a noncytotoxic and relevant occupational exposure concentration) and LC50 PFOA ex vivo. This perfluorinated compound also caused the blockage of GJIC in cumulus cells-oocyte complexes (COCs) obtained from female mice exposed in vivo, as evaluated by calcein transfer from cumulus cells to the oocyte. The ability of PFOA of disrupting the GJIC in COCs, generating ROS in the fetal ovary and causing apoptosis and necrosis in mammal's oocytes, might account for the reported association between increasing maternal plasma concentrations of PFOA with reduced fertility in women.

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OBJECTIVE: The number of successful pregnancies in kidney transplant (KT) recipients has increased in recent years. Little evidence is available about the risk of in utero immunosuppressive exposure for long-term cognitive consequences. The aim of this study was to evaluate the impact of immunosuppression during pregnancy on intellectual performance of children born to KT recipients. METHODS: Using a cross-sectional design, women who had undergone KT and their children (aged 4+ years) were recruited at the outpatient follow-up in five transplant centers. Women who did not receive immunosuppression during pregnancy with similar distributions of socioeconomic status and length of gestation and their children were also recruited. Children were assessed with Wechsler Intelligence Scales. RESULTS: The study sample included 50 exposed and 50 unexposed children. No differences between groups in all the proposed confounding factors were found. Full-scale IQ did not differ significantly between both groups. Also, significant differences in any index or subscale score were not observed, with the exception of time required to complete the Wechsler preschool and primary scale of intelligence (WPPSI) Zoo locations subtest, which was done quicker in the unexposed group (p = .007). Exposure to immunosuppression during pregnancy was not a significant predictor of low IQ in logistic regression after adjustment for other factors. CONCLUSIONS: Immunosuppression therapy during pregnancy of KT women did not affect global intellectual performance of their offspring, except maybe for visuospatial working memory in preschool children.

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Introducción: En la actualidad la litiasis renal constituye entre el 20 y el 30 por ciento de las consultas de urología en el mundo. Objetivos: Identificar las complicaciones intra- y posoperatorias de la nefrolitotomía percutánea en pacientes con litiasis coraliforme. Métodos: Se realizó un estudio descriptivo en pacientes con diagnóstico de litiasis coraliforme ingresados para nefrolitotomía percutánea electiva en el Hospital Clínico Quirúrgico Hermanos Ameijeiras entre octubre 2010 y entre octubre 2015. Resultados: Se identificaron 32 pacientes con complicaciones intra y posoperatorias. Del sexo masculino fueron 68,7 por ciento. La media de la edad fue 47,59 ± 12,2 años. La clasificación ASA más frecuente fue ASA II en 56,2 por ciento. Todos los pacientes recibieron anestesia general. El valor medio de pérdidas hemáticas intraoperatorias fue de 650,00 ± 413,09 mL. La reposición total de volumen fue de 10, 107,80 ± 2, 659,25 mL. Se administró cloro sodio al 0,9 por ciento 7743,75 ± 2007,39 mL y concentrado de hematíes a 18,8 por ciento del total. Las complicaciones intraoperatorias se presentaron en 24 pacientes 75,0 por ciento. De ellas las más frecuentes fueron las metabólicas, cardiovasculares, respiratorias y renales. La hipotermia estuvo presente en la tercera parte de los enfermos. Las complicaciones posoperatorio se presentaron en 23 pacientes 71,9 por ciento. De ellas, cardiovasculares, respiratorias, renales y sépticas fueron las más frecuentes. Conclusiones: Las complicaciones fueron frecuentes y graves; sin embargo, se logró un porcentaje importante de pacientes egresados curados(AU)

Introduction: Renal lithiasis currently accounts for 20-30 percent of urology consultations worldwide. Objectives: To identify the intra- and postoperative complications of percutaneous nephrolithotomy in patients with staghorn lithiasis. Methods: A descriptive study was conducted in patients with a diagnosis of staghorn lithiasis and admitted for elective percutaneous nephrolithotomy Hermanos Ameijeiras Clinical-Surgical Hospital between October 2010 and October 2015. Results: We identified 32 patients with intra- and postoperative complications. The male sex was represented by the 68.7 percent. The mean age was 47.59±12.2 years. The most frequent ASA classification was ASA II, accounting for 56.2 percent. All patients received general anesthesia. The mean value of intraoperative hematic losses was 650.00±413.09 mL. Total volume replacement was 10, 107.80±2, 659.25 mL. Chloride sodium 0.9 percent was administered at doses 7743.75±2007.39 mL and packed red blood cells to 18.8 percent of the total. Intraoperative complications occurred in 24 patients, accounting for 75.0 percent. Of these, the most frequent were metabolic, cardiovascular, respiratory and renal. Hypothermia was present in one third of the patients. Postoperative complications occurred in 23 patients, accounting for 71.9 percent. Of these, cardiovascular, respiratory, renal and septic were the most frequent. Conclusions: The complications were frequent and serious. However, an important percentage number was achieved for cured discharge patients(AU)

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BACKGROUND Real-time reverse transcription polymerase chain reaction (RT-PCR) is routinely used to detect viral infections. In Brazil, it is mandatory the use of nucleic acid tests to detect hepatitis C virus (HCV), hepatitis B virus and human immunodeficiency virus in blood banks because of the immunological window. The use of an internal control (IC) is necessary to differentiate the true negative results from those consequent from a failure in some step of the nucleic acid test. OBJECTIVES The aim of this study was the construction of virus-modified particles, based on MS2 bacteriophage, to be used as IC for the diagnosis of RNA viruses. METHODS The MS2 genome was cloned into the pET47b(+) plasmid, generating pET47b(+)-MS2. MS2-like particles were produced through the synthesis of MS2 RNA genome by T7 RNA polymerase. These particles were used as non-competitive IC in assays for RNA virus diagnostics. In addition, a competitive control for HCV diagnosis was developed by cloning a mutated HCV sequence into the MS2 replicase gene of pET47b(+)-MS2, which produces a non-propagating MS2 particle. The utility of MS2-like particles as IC was evaluated in a one-step format multiplex real-time RT-PCR for HCV detection. FINDINGS We demonstrated that both competitive and non-competitive IC could be successfully used to monitor the HCV amplification performance, including the extraction, reverse transcription, amplification and detection steps, without compromising the detection of samples with low target concentrations. In conclusion, MS2-like particles generated by this strategy proved to be useful IC for RNA virus diagnosis, with advantage that they are produced by a low cost protocol. An attractive feature of this system is that it allows the construction of a multicontrol by the insertion of sequences from more than one pathogen, increasing its applicability for diagnosing different RNA viruses.

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BACKGROUND: Real-time reverse transcription polymerase chain reaction (RT-PCR) is routinely used to detect viral infections. In Brazil, it is mandatory the use of nucleic acid tests to detect hepatitis C virus (HCV), hepatitis B virus and human immunodeficiency virus in blood banks because of the immunological window. The use of an internal control (IC) is necessary to differentiate the true negative results from those consequent from a failure in some step of the nucleic acid test. OBJECTIVES: The aim of this study was the construction of virus-modified particles, based on MS2 bacteriophage, to be used as IC for the diagnosis of RNA viruses. METHODS: The MS2 genome was cloned into the pET47b(+) plasmid, generating pET47b(+)-MS2. MS2-like particles were produced through the synthesis of MS2 RNA genome by T7 RNA polymerase. These particles were used as non-competitive IC in assays for RNA virus diagnostics. In addition, a competitive control for HCV diagnosis was developed by cloning a mutated HCV sequence into the MS2 replicase gene of pET47b(+)-MS2, which produces a non-propagating MS2 particle. The utility of MS2-like particles as IC was evaluated in a one-step format multiplex real-time RT-PCR for HCV detection. FINDINGS: We demonstrated that both competitive and non-competitive IC could be successfully used to monitor the HCV amplification performance, including the extraction, reverse transcription, amplification and detection steps, without compromising the detection of samples with low target concentrations. In conclusion, MS2-like particles generated by this strategy proved to be useful IC for RNA virus diagnosis, with advantage that they are produced by a low cost protocol. An attractive feature of this system is that it allows the construction of a multicontrol by the insertion of sequences from more than one pathogen, increasing its applicability for diagnosing different RNA viruses.

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Introduction: Valved conduits for surgically reconstructing the outflow tract of the right ventricle have improved the prognosis for certain congenital heart diseases. When they become dysfunctional, transcatheter pulmonary valve replacement is safe and effective. Clinical findings, diagnostic evaluation and interventions: We report the case of a 21 years old female patient diagnosed with transposition of great vessels corrected in childhood. After several surgeries, she presents a dysfunction of the right ventricle. In response, a "Melody " pulmonary valve was implanted percutaneously under deep sedation. Conclusion: The ideal anesthetic technique for transcatheter pulmonary valve replacement is controversial. There is a debate over the merits of general anesthesia and deep sedation. Each case must be decided on after a careful preoperative evaluation, considering the risk/benefit and patient cooperation. The experience at our center using deep sedation is promising, but we have an alternative plan to convert to general anesthesia if necessary.

Introducción: Los conductos valvulados para reconstruir quirúrgicamente el tracto de salida del ventrículo derecho han mejorado el pronóstico de ciertas cardiopatía congénitas. Cuando éstos se vuelven disfuncionantes, el remplazo valvular pulmonar transcatéter es eficaz y seguro. Hallazgos clínicos, evaluación diagnóstica e intervenciones: Presentamos una paciente de 21 años, diagnosticada de transposición de grandes vasos corregida en la infancia, que tras varias intervenciones quirúrgicas presenta una disfunción del conducto del ventrículo derecho, por lo que se implanta percutáneamente una válvula pulmonar "Melody" bajo sedación profunda. Conclusión: La técnica anestésica ideal para el recambio valvular transcatéter pulmonar es controvertida, discutiéndose entre la anestesia general y la sedación profunda. Cada caso debe decidirse tras una evaluación preoperatoria cuidadosa, considerando el riesgo/beneficio y la colaboración del paciente. La experiencia en nuestro centro empleando la sedación profunda es prometedora, aunque debemos contar con un plan alternativo para reconvertir a anestesia general de ser necesario.

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Objetivos Estudios concluyen que la máxima tensión y distribución de fuerzas se produciría alrededor del cuello del implante. Comparar la distribución de las tensiones entre un implante dental corto oseointegrado en distintas disponibilidades óseas verticales y un implante estándar oseointegrado en el sector posterior del maxilar en un terreno mixto formado por hueso propio del paciente y Bio-Oss®. Conocer la distribución de las tensiones. Estudiar el aumento del diámetro del implante corto. Analizar si los resultados avalan el uso de implantes cortos. Material y método El método de elementos finitos (MEF), que permite resolver ecuaciones diferenciales asociadas a un problema físico sobre geometrías complicadas. En este trabajo la región geométrica es un modelo tridimensional de un implante, su corona, y una porción de la región ósea de la zona estudiada. Los modelos fueron sometidos a fuerzas de oclusión de 150 N en ángulo de 30° Norma ISO 14801:2003. El software de MEF que se utiliza es el ABAQUS de la empresa Dassault Systèmes. Resultados Máximos valores se concentran en la porción cervical del implante. Las tensiones en el implante están dentro del mismo rango. A mayor módulo de elasticidad de los elementos que componen los modelos, mayor es la absorción de las fuerzas. Las tensiones en el hueso cortical no mostraron diferencias, pero en el modelo que aumentamos el diámetro del implante a 4,8 se produce una marcada disminución de las tensiones. La comparativa de las tensiones en el hueso esponjoso muestra que existe diferencia en las tensiones producidas en el hueso con Bio-Oss® y está localizado en la porción apical del implante quedando lejos de la zona de mayor concentración de los esfuerzos. Conclusiones La máxima concentración de las fuerzas a nivel cervical es independiente de la longitud del implante, siendo más favorable el aumento del diámetro. El uso de implantes cortos en hueso de baja calidad parece posible.

Objectives Current studies conclude that the maximum tension and the greater distribution of forces should occur around the implant neck. To compare the distribution of stress between a short dental implant osseointegrated in different available vertical bones and standard osseointegrated implants in the posterior maxilla in mixed terrain formed by the bone of the patient and Bio-Oss®. To determine the stress distribution. To study the increased diameter of the short implant. To determine whether the results support the use of short implants. Materials and methods The finite elements method (FEM), which helps to solve differential equations associated with a physical problem with complicated geometries, was used in this work, where the geometric region is a three-dimensional model of an implant, its crown, and a portion of the bone region of the studied area. The models were subjected to occlusion forces, 150 N Angle 30° ISO 14801: 2003. The MEF software used was called Abaqus from Dasssault Systemes Enterprise. Results The maximum values were concentrated in the cervical portion of the implant. Tensions in the implant are in the same range. The greater the elasticity of the elements contained in the module, the greater is the absorption of stress forces. The tension in the cortical bone showed no differences, but in the model where the diameter of the implant is increased to 4.8, a marked decrease occurs in the bone stress. The comparison of the stresses in the cancellous bone showed a difference in the stresses produced in the bone with Bio-Oss®, and it is located in the apical portion of the implant away from the area of the major stress concentration. Conclusions The maximum concentration of forces in cervical portion is independent of the length of the implant, being favourable to increase the diameter. It is possible to use low quality bone in for shorts implants.

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Cell therapy has frequently been reported as a possible treatment for spinal trauma in humans and animals; however, without pharmacologically curative action on damage from the primary lesion. In this study, we evaluated the effect of administering human adipose-derived stem cells (hADSC) in rats after spinal cord injury. The hADSC were used between the third and fifth passages and a proportionof cells were transduced for screening in vivo after transplantation. Spinal cord injury was inducedwith a Fogarty catheter no. 3 inserted into the epidural space with a cuff located at T8 and filled with 80-μL saline for 5 min. The control group A (n = 12) received culture medium (50 μL) and group B (n = 12) received hADSC (1.2 × 106) at 7 and 14 days post-injury, in the tail vein. Emptying of thebladder by massage was performed daily for 3 months. Evaluation of functional motor activity was performed daily until 3 months post-injury using the Basso-Beattie-Bresnahan scale. Subsequently, the animals were euthanized and histological analysis of the urinary bladder and spinal cord was performed. Bioluminescence analysis revealed hADSC at the application site and lungs. There was improvement of urinary bladder function in 83.3% animals in group B and 16.66% animals in group A. The analysis of functional motor activity and histology of the spinal cord and urinary bladder demonstrated nosignificant difference between groups A and B. The results indicate that transplanted hADSC improved urinary function via a telecrine mechanism, namely action at a distance.(AU)

A terapia celular tem sido reportada como possibilidade de tratamento para os traumas medulares, frequentes em pessoas e em animais, e que não apresentam cura farmacológica para danos advindos da lesão primária. Foram avaliados os efeitos da aplicação de células-tronco derivadas do tecido adiposo humano (hADSC) em ratos Wistar após lesão medular. As hADSC foram cultivadas, utilizadas entre aterceira e quinta passagens e parte delas foi transduzida para o rastreamento in vivo após o transplante. A lesão medular foi realizada com cateter de Fogarty nº. 3, introduzido no espaço epidural, com cuff localizado em T8 e preenchido com 80 μL de solução salina, por 5 minutos. Os animais do grupocontrole (A, n=12) receberam aplicações de meio de cultura (50 μL) e o grupo B (n=12) recebeu duas aplicações de hADSC (1,2 x 106), sete e 14 dias pós-lesão, ambas na veia da cauda. O esvaziamento vesical, por massagem, foi efetuado diariamente, durante três meses. As avaliações motoras foramrepetidas, diariamente, até três meses pós-lesão, utilizando a escala Basso-Beattie-Bresneham. Após este período os animais foram submetidos à eutanásia e foram realizadas análises histológicas de bexigaurinária e medula espinhal. As análises de bioluminescência revelaram hADSC no local da aplicação e nos pulmões. Houve melhora da função da bexiga urinária em 83,3% dos animais do grupo B e 16,66% dos animais do grupo A. A análise das avaliações motoras e histológicas das medulas espinhais e bexigas urinárias não demonstrou diferenças significativas entre os grupos A e B. Os resultados indicam que as hADSC transplantadas influenciaram a micção, de forma benéfica, por um mecanismo de açãoà distância, telécrino.(AU)

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UNLABELLED: Stem cell-based regenerative medicine is one of the most intensively researched medical issues. Pre-clinical studies in a large-animal model, especially in swine or miniature pigs, are highly relevant to human applications. Mesenchymal stem cells (MSCs) have been isolated and expanded from different sources. OBJECTIVE: This study aimed at isolating and characterizing, for the first time, bone marrow-derived MSCs (BM-MSCs) from a Brazilian minipig (BR1). Also, this aimed to validate a new large-animal model for stem cell-based tissue engineering. MATERIAL AND METHODS: Bone marrow (BM) was aspirated from the posterior iliac crest of twelve adult male BR1 under general anesthesia. MSCs were selected by plastic-adherence as originally described by Friedenstein. Cell morphology, surface marker expression, and cellular differentiation were examined. The immunophenotypic profile was determined by flow cytometry. The differentiation potential was assessed by cytological staining and by RT-PCR. RESULTS: MSCs were present in all minipig BM samples. These cells showed fibroblastic morphology and were positive for the surface markers CD90 (88.6%), CD29 (89.8%), CD44 (86.9%) and negative for CD34 (1.61%), CD45 (1.83%), CD14 (1.77%) and MHC-II (2.69%). MSCs were differentiated into adipocytes, osteoblasts, and chondroblasts as demonstrated by the presence of lipidic-rich vacuoles, the mineralized extracellular matrix, and the great presence of glycosaminoglycans, respectively. The higher gene expression of adipocyte fatty-acid binding protein (AP2), alkaline phosphatase (ALP) and collagen type 2 (COLII) also confirmed the trilineage differentiation (p<0.001, p<0.001, p=0.031; respectively). CONCLUSIONS: The isolation, cultivation, and differentiation of BM-MSCs from BR1 makes this animal eligible as a useful large-animal model for stem cell-based studies in Brazil.

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Stem cell-based regenerative medicine is one of the most intensively researched medical issues. Pre-clinical studies in a large-animal model, especially in swine or miniature pigs, are highly relevant to human applications. Mesenchymal stem cells (MSCs) have been isolated and expanded from different sources. Objective: This study aimed at isolating and characterizing, for the first time, bone marrow-derived MSCs (BM-MSCs) from a Brazilian minipig (BR1). Also, this aimed to validate a new large-animal model for stem cell-based tissue engineering. Material and Methods: Bone marrow (BM) was aspirated from the posterior iliac crest of twelve adult male BR1 under general anesthesia. MSCs were selected by plastic-adherence as originally described by Friedenstein. Cell morphology, surface marker expression, and cellular differentiation were examined. The immunophenotypic profile was determined by flow cytometry. The differentiation potential was assessed by cytological staining and by RT-PCR. Results: MSCs were present in all minipig BM samples. These cells showed fibroblastic morphology and were positive for the surface markers CD90 (88.6%), CD29 (89.8%), CD44 (86.9%) and negative for CD34 (1.61%), CD45 (1.83%), CD14 (1.77%) and MHC-II (2.69%). MSCs were differentiated into adipocytes, osteoblasts, and chondroblasts as demonstrated by the presence of lipidic-rich vacuoles, the mineralized extracellular matrix, and the great presence of glycosaminoglycans, respectively. The higher gene expression of adipocyte fatty-acid binding protein (AP2), alkaline phosphatase (ALP) and collagen type 2 (COLII) also confirmed the trilineage differentiation (p<0.001, p<0.001, p=0.031; respectively). Conclusions: The isolation, cultivation, and differentiation of BM-MSCs from BR1 makes this animal eligible as a useful large-animal model for stem cell-based studies in Brazil. .

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Cell therapy has frequently been reported as a possible treatment for spinal trauma in humans and animals; however, without pharmacologically curative action on damage from the primary lesion. In this study, we evaluated the effect of administering human adipose-derived stem cells (hADSC) in rats after spinal cord injury. The hADSC were used between the third and fifth passages and a proportionof cells were transduced for screening in vivo after transplantation. Spinal cord injury was inducedwith a Fogarty catheter no. 3 inserted into the epidural space with a cuff located at T8 and filled with 80-μL saline for 5 min. The control group A (n = 12) received culture medium (50 μL) and group B (n = 12) received hADSC (1.2 × 106) at 7 and 14 days post-injury, in the tail vein. Emptying of thebladder by massage was performed daily for 3 months. Evaluation of functional motor activity was performed daily until 3 months post-injury using the Basso-Beattie-Bresnahan scale. Subsequently, the animals were euthanized and histological analysis of the urinary bladder and spinal cord was performed. Bioluminescence analysis revealed hADSC at the application site and lungs. There was improvement of urinary bladder function in 83.3% animals in group B and 16.66% animals in group A. The analysis of functional motor activity and histology of the spinal cord and urinary bladder demonstrated nosignificant difference between groups A and B. The results indicate that transplanted hADSC improved urinary function via a telecrine mechanism, namely action at a distance.

A terapia celular tem sido reportada como possibilidade de tratamento para os traumas medulares, frequentes em pessoas e em animais, e que não apresentam cura farmacológica para danos advindos da lesão primária. Foram avaliados os efeitos da aplicação de células-tronco derivadas do tecido adiposo humano (hADSC) em ratos Wistar após lesão medular. As hADSC foram cultivadas, utilizadas entre aterceira e quinta passagens e parte delas foi transduzida para o rastreamento in vivo após o transplante. A lesão medular foi realizada com cateter de Fogarty nº. 3, introduzido no espaço epidural, com cuff localizado em T8 e preenchido com 80 μL de solução salina, por 5 minutos. Os animais do grupocontrole (A, n=12) receberam aplicações de meio de cultura (50 μL) e o grupo B (n=12) recebeu duas aplicações de hADSC (1,2 x 106), sete e 14 dias pós-lesão, ambas na veia da cauda. O esvaziamento vesical, por massagem, foi efetuado diariamente, durante três meses. As avaliações motoras foramrepetidas, diariamente, até três meses pós-lesão, utilizando a escala Basso-Beattie-Bresneham. Após este período os animais foram submetidos à eutanásia e foram realizadas análises histológicas de bexigaurinária e medula espinhal. As análises de bioluminescência revelaram hADSC no local da aplicação e nos pulmões. Houve melhora da função da bexiga urinária em 83,3% dos animais do grupo B e 16,66% dos animais do grupo A. A análise das avaliações motoras e histológicas das medulas espinhais e bexigas urinárias não demonstrou diferenças significativas entre os grupos A e B. Os resultados indicam que as hADSC transplantadas influenciaram a micção, de forma benéfica, por um mecanismo de açãoà distância, telécrino.

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Over the last years an expanding family of small RNAs (i.e. microRNAs, siRNAs and piRNAs) was recognized as key players in diverse forms of gene silencing and chromatin organization. Effectors functions of these small RNAs are achieved through ribonucleoprotein (RNP) complexes containing at their center an Argonaute/Piwi protein. Although these proteins and their small RNA-associated machinery can be traced back to the common ancestor of eukaryotes, this machinery seems to be entirely lost or extensively simplified in some unicellular organisms including Trypanosoma cruzi, which are unable to trigger RNAi related phenomena. Speculating about the presence of alternate small RNA-mediated pathways in these organisms, we constructed and analyzed a size-fractionated cDNA library (20-35 nt) from epimastigotes forms of T. cruzi. Our results showed the production of an abundant class of tRNA-derived small RNAs preferentially restricted to specific isoacceptors and whose production was more accentuated under nutritional stress. These small tRNAs derived preferentially from the 5' halves of mature tRNAs and were recruited to distinctive cytoplasmic granules. Our data favor the idea that tRNA cleavage is unlikely to be the consequence of non-specific degradation but a controlled process, whose biological significance remains to be elucidated.

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Se estudiaron 6 548 pacientes que asistieron al cuerpo de guardia del Instituto Superior de Medicina Militar “Dr Luis Díaz Soto”, con diagnóstico de traumatismo craneoencefálico, independientemente de su grado de gravedad y de la existencia de lesiones múltiples asociadas. De estos, el 19,62 por ciento correspondió a niños. La causa más frecuente fue el accidente del tránsito. Predominó el traumatismo craneoencefálico ligero y solo 383 pacientes se clasificaron entre moderado y severo. El 18,35 por ciento de los casos requirió ingreso hospitalario y el 4,54 por ciento demandó algún tipo de intervención quirúrgica de urgencia. En los pacientes con traumatismo craneoencefálico grave se obtuvo una mortalidad del 42 por ciento, y en los enfermos operados del 27,61 por ciento. La complicación más frecuente fue la sepsis respiratoria. Se obtuvo buenos resultados en el 47,71 por ciento de los casos con traumatismo craneoencefálico severo y de forma general en el 98,41 por ciento de los pacientes estudiados

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Se realizó un estudio prospectivo, descriptivo y longitudinal de la conducta neuroquirúrgica seguida con todos los pacientes atendidos desde enero de 1998 hasta diciembre de 2002 en el Cuerpo de Guardia del Instituto Superior de Medicina Militar “Dr. Luis Díaz Soto”. Los datos se obtuvieron en las entregas de guardia, registros médicos, historias clínicas, libro de control de intervenciones quirúrgicas y de ingresos. En total se atendieron 8 113 pacientes, de ellos el 17 por ciento correspondió a niños. Requirieron ingresos 1 357 enfermos; las fracturas de cráneo constituyeron el principal motivo (254 casos), seguidas por la conmoción cerebral (245 casos). Se intervinieron quirúrgicamente 525 pacientes, por causa traumática 285; los hematomas intracraneales con 167 pacientes constituyeron la primera causa de intervención quirúrgica. Se hizo un análisis de las causas de ingreso, remisiones e intervenciones neuroquirúrgicas. La traumatología de cráneo sigue siendo la principal causa de atención neuroquirúrgica de urgencia, aunque aún el especialista de guardia ve a un gran número de pacientes que no requieren atención especializada

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