Analysis of the antigenic and prophylactic properties of the Leishmania translation initiation factors eIF2 and eIF2B in natural and experimental leishmaniasis

Los diferentes miembros de las familias de proteínas intracelulares son reconocidos por el sistema inmunológico del huésped vertebrado infectado por parásitos del género Leishmania. Aquí hemos analizado las propiedades antigénicas e inmunogénicas de los factores de iniciación de la traducción de Leishmania eIF2 y eIF2B. Una búsqueda in silico en las bases de datos de secuencias de Leishmania infantum permitió la identificación de los genes que codifican las subunidades α, β y γ y las subunidades α, β y δ de los supuestos ortólogos de Leishmania de los factores de iniciación eucarióticos F2 (LieIF2) o F2B (LieIF2B), respectivamente. La antigenicidad de estos factores fue analizada por ELISA utilizando versiones recombinantes de las diferentes subunidades. Se encontraron anticuerpos contra las diferentes subunidades LieIF2 y LieIF2B en los sueros de pacientes con leishmaniosis visceral humana y canina, y también en los sueros de hámsteres infectados experimentalmente con L. infantum. En ratones desafiados por L. infantum (BALB/c) y Leishmania major (BALB/c o C57BL/6) se detectó una respuesta humoral moderada contra estos factores proteicos. Cabe destacar que estas proteínas provocaron una producción de IL-10 por parte de los esplenocitos derivados de ratones infectados, independientemente de la especie de Leishmania empleada para el desafío experimental. Cuando se administraron vacunas de ADN basadas en la expresión de los genes codificantes de las subunidades LieIF2 o LieIF2B en ratones, se observó una secreción específica de antígenos de citoquinas IFN-γ e IL-10. Además, se generó en los ratones vacunados una protección parcial contra el desarrollo de la CL murina debido a la infección por L. major. Además, en este trabajo mostramos que la subunidad LieIF2α y las subunidades LieIF2Bβ y δ tienen la capacidad de estimular la secreción de IL-10 por las células del bazo de los ratones ingenuos. Los linfocitos B fueron identificados como los mayores productores de esta citoquina antiinflamatoria. Teniendo en cuenta los datos encontrados en este estudio, se puede formular la hipótesis de que estas proteínas actúan como factores de virulencia implicados en la inducción de respuestas humorales, así como en la producción de la citoquina IL-10 de regulación descendente, favoreciendo un resultado patológico. Por lo tanto, estas proteínas podrían considerarse marcadores de enfermedad.Different members of intracellular protein families are recognized by the immune system of the vertebrate host infected by parasites of the genus Leishmania. Here, we have analyzed the antigenic and immunogenic properties of the Leishmania eIF2 and eIF2B translation initiation factors. An in silico search in Leishmania infantum sequence databases allowed the identification of the genes encoding the α, β, and γ subunits and the α, β, and δ subunits of the putative Leishmania orthologs of the eukaryotic initiation factors F2 (LieIF2) or F2B (LieIF2B), respectively. The antigenicity of these factors was analyzed by ELISA using recombinant versions of the different subunits. Antibodies against the different LieIF2 and LieIF2B subunits were found in the sera from human and canine visceral leishmaniasis patients, and also in the sera from hamsters experimentally infected with L. infantum. In L. infantum (BALB/c) and Leishmania major (BALB/c or C57BL/6) challenged mice, a moderate humoral response against these protein factors was detected. Remarkably, these proteins elicited an IL-10 production by splenocytes derived from infected mice independently of the Leishmania species employed for experimental challenge. When DNA vaccines based on the expression of the LieIF2 or LieIF2B subunit encoding genes were administered in mice, an antigen-specific secretion of IFN-γ and IL-10 cytokines was observed. Furthermore, a partial protection against murine CL development due to L. major infection was generated in the vaccinated mice. Also, in this work we show that the LieIF2α subunit and the LieIF2Bβ and δ subunits have the capacity to stimulate IL-10 secretion by spleen cells from naïve mice. B-lymphocytes were identified as the major producers of this anti-inflammatory cytokine. Taking into account the data found in this study, it may be hypothesized that these proteins act as virulence factors implicated in the induction of humoral responses as well as in the production of the down-regulatory IL-10 cytokine, favoring a pathological outcome. Therefore, these proteins might be considered markers of disease.• Ministerio de Ciencia e Innovación y Fondos FEDER. Proyectos FISPI14/00366, FISPI14/00366 • Fondo de Investigaciones Sanitarias. Proyecto ISCIII-RETICRD16/0027/0008-FEDER • Conselho Nacional de Desenvolvimento Científico e Tecnológico (Brasil). Program 300174/2014-4 • Centro de Biología Molecular Severo Ochoa. Ayuda • Fundación Ramón Areces. Ayuda • Banco de Santander. AyudapeerReviewe

Outcomes of ureteroscopy miniaturization on tissue damage and tissue hypoxia in a pig model

Miniaturization of ureteroscopy materials is intended to decrease tissue damage. However, tissue hypoxia and the gross and microscopic effects on tissue have not been adequately assessed. We compared the gross and microscopic effects of micro-ureteroscopy (m-URS) and conventional ureteroscopy (URS) on the urinary tract. We employed 14 pigs of the Large White race. URS was performed in one of the ureters with an 8/9.8 F ureteroscope, while a 4.85 F m-URS sheath was used in the contralateral ureter. Gross assessment of ureteral wall damage and ureteral orifice damage was performed. For microscopic assessment hematoxylin-eosin staining and immunohistochemistry for detection of tissue hypoxia were conducted. Regarding the macroscopic assessment of ureteral damage, substantial and significant differences were recorded using URS (C = 0.8), but not with m-URS. Microscopic assessment after staining with hematoxylin-eosin revealed greater epithelial desquamation in the URS group (p < 0.05). Pimonidazole staining revealed greater hypoxia in the epithelial cells than in the remainder of the ureteral layers. We conclude that m-URS causes less damage to the ureteral orifice than URS. Histopathological findings show m-URS reduces ureteral epithelial damage compared with conventional ureteroscopy. Both URS and m-URS cause cellular hypoxia.The present study is funded by the Alicante Institute for Health and Biomedical Research (ISABIAL-FISABIO Foundation) and Presurgy S.L

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

Manual de simulación clínica en especialidades médicas

Manual sobre técnicas y modos de simulación clínica en diversas especialidades médicas.La enseñanza y formación en medicina necesita el uso de la simulación. Existen evidencias de su uso desde hace cientos de años, pero, en los últimos años se ha incrementado y diseminado. La simulación clínica está validada científicamente en múltiples contextos médicos y de otras áreas profesionales de la salud. Y es considerada de gran importancia como proceso de entrenamiento y de mejora de las competencias y adquisición de habilidades médicas en campos que incluye desde la historia clínica, comunicación con el paciente, exploración, diagnóstico terapéutica médica-farmacológica y quirúrgica y seguridad al tratar al paciente. Hoy en día, para muchas técnicas y situaciones clínicas es inaceptable llegar junto a los pacientes sin un dominio adquirido en simulación. La simulación puede ocurrir sin el uso de recursos adicionales, solo las personas, o utilizando pocos o muchos recursos de baja hasta alta tecnología y se puede adaptar a los recursos disponibles, abarcando todas las áreas de conocimiento, y dentro de ellas competencias técnicas o actitudes, solas o en conjunto. El uso racional y basado en evidencia de la simulación es de la mayor importancia por la necesidad de una mayor efectividad y eficiencia en la transformación de los profesionales de la salud para que puedan mejorar su capacidad de atender a los pacientes. La simulación es también una buena herramienta de evaluación de competencias y habilidades en Medicina y otras disciplinas de las Ciencias de la Salud Este manual incluye técnicas y modos de simulación clínica en diversas especialidades médicas, útiles, para quien busque un manual práctico y actualizado.Cátedra de Mecenazgo de la Universidad de Málaga. Cátedra de Terapias Avanzadas en Patología Cardiovascular Cátedra de Mecenazgo de la Universidad de Málaga. Cátedra de Investigación Biomédica Quirón Salu

Estudio experimental comparativo de nuevos tratamientos endourológicos de la uropatía obstructiva mediante stents metálicos recubiertos

Tesis doctoral con la Mención de "Doctor Internacional"El manejo de la obstrucción ureteral de origen maligno o de carácter benigno complicado continúa siendo a día de hoy un desafío para la comunidad urológica. La disposición de catéteres ureterales doble jota como alternativa de derivación interna y de la nefrostomía percutánea como derivación externa, son las medidas terapéuticas que con mayor frecuencia se emplean en este tipo de pacientes. Sin embargo, la morbilidad asociada a ambas derivaciones provoca que sean necesarias otras vías de tratamiento. En este sentido, el empleo de stents metálicos podría satisfacer las expectativas, ya que permiten en un solo acto la resolución de la estenosis, sin derivaciones externas, y no se asocian con ninguno de los efectos adversos de las derivaciones urinarias actuales, y tampoco necesitan recambiar los catéteres ni la sonda de nefrostomía, lo que disminuye la calidad de vida de sus portadores. Sin embargo, a pesar de estas buenas perspectivas de éxito, cimentadas en los buenos resultados sobre todo de los stents vasculares, los stents ureterales no cumplen las perspectivas. La migración, la incrustación y sobre todo el crecimiento hiperplásico urotelial son las principales causas de que estos dispositivos no sean plenamente aceptados. El presente trabajo experimental en modelo porcino pretende evaluar la Hipótesis de que la alteración de la interacción entre los extremos del stent metálico, la pared ureteral y el peristaltismo ureteral disminuye la aparición de la reacción hiperplásica urotelial en los extremos del stent, lo que puede favorecer la expansión de los stents en el tracto urinario superior. Para ello se distribuyen los 60 animales del estudio en cinco grupos diferentes todos ellos con un modelo de estenosis ureteral experimental. Lo que permite evaluar diferentes técnicas diseñadas para disminuir o inhibir la formación de hiperplasia urotelial tras la disposición de un stent metálico. La primera técnica a evaluar es la realización de una endoureteromía en ambos extremos del stent, para de este modo alterar el peristaltismo en la zona de confluencia entre el stent y el uréter sano. Otro posibilidad terapéutica evaluada es la disposición de un novedoso diseño de stent ureteral híbrido y dinámico que no afecta al peristaltismo del uréter adyacente al stent y por ende, no debe producir hiperplasia. Por último, se evalúa en este estudio si la disposición coadyuvante de un catéter ureteral doble jota junto a un stent metálico ureteral disminuye la formación de hiperplasia urotelial. Los resultados del estudio experimental avalan la hipótesis de trabajo. Los grupos de estudio frente a sus respectivos grupos control evidencian que, la endoureterotomía en los extremos del stent disminuye significativamente el desarrollo de hiperplasia. Asimismo el nuevo diseño de stent metálico ureteral, HMS, es seguro, y efectivo en el tratamiento de la estenosis ureteral disminuyendo la formación de hiperplasia urotelial sin la necesidad de manipulación secundaria para asegurar la permeabilidad del stent. Por último, la disposición de un catéter ureteral coaxial al stent no disminuye la aparición de hiperplasia, pero sí su agresividad significativamente. Por todo ello, podemos concluir que la formación de hiperplasia urotelial relacionada con la disposición de stents ureterales metálicos disminuye significativamente cuando se reduce la interacción entre los extremos del stent y el ureter peristáltico adyacente. Además de que los catéteres doble jota no inhiben la formación de hiperplasia al asociarlos con los stents ureterales metálicos. De las tres terapias evaluadas, el nuevo stent ureteral, HMS, es el que mejores resultados alcanza, mínimos efectos adversos muestra y es el que menor manipulación ureteral provoca.The management of a malignant or complicated benign ureteral obstruction still today represents a challenge for the urological community. The placement of double J ureteral catheters as an alternative to internal diversion, and percutaneous nephrostomy as an external diversion, are the most frequently used therapeutic measures for this type of patient. However, morbidity associated with both diversions makes other treatment options necessary. In this sense, the use of metal stents could meet the expectations, as they allow stenosis to be solved in a single act, with no external diversions. In addition, they are not associated with any of the adverse effects caused by present urinary diversions and their catheters or nephrostomy catheter do not need to be replaced, which reduces carriers' quality of life. However, in spite of successful prospects, based on the positive results especially shown by vascular stents, ureteral stents do not meet our expectations. Migration, encrustation and especially urothelial hyperplasic growth are the main reasons why these devices are not wholly accepted. The present experimental study on swine model aims to assess the hypothesis that alteration in the interaction of the metal stent ends, ureteral wall and ureteral peristalsis reduces the onset of urothelial hyperplasic reaction at stent ends. This can favor stent expansion in the upper urinary tract. The 60 study animals are divided into five different groups, all of them having an experimental ureteral stenosis model. In this way, the different techniques designed to reduce or inhibit urothelial hyperplasia formation after metal stent placement are assessed. The first technique to be assessed is the performance of an endoureterotomy at both stent ends, so that peristalsis can be altered in the convergence area between stent and healthy ureter. Another therapeutic possibility assessed is to place an innovative design for a hybrid and dynamic ureteral stent, which does not involve the ureter peristalsis adjacent to the stent, and therefore should not cause hyperplasia. Finally, this study assesses whether coadjuvant placement of a double J ureteral catheter, together with a ureteral metal stent, reduces urothelial hyperplasia formation. Experimental study results support the working hypothesis. Study groups compared to their respective control groups prove that endoureterotomy at stent ends significantly reduces hyperplasia development. Likewise HMS, the new ureteral metal stent design, is safe and effective for the treatment of ureteral stenosis, as it decreases urothelial hyperplasia formation with no need for a secondary manipulation in order to ensure stent permeability. Finally, placing a ureteral catheter coaxially to the stent does not reduce hyperplasia onset. However, it does significantly reduce its aggressivity. Therefore, we can state that urothelial hyperplasia formation related to metal ureteral stent placement significantly decreases when interaction between stent ends and adjacent peristaltic ureter is reduced. In addition, double J catheters do not inhibit hyperplasia formation when they are associated with metal ureteral stents. Among the three therapies assessed, HMS, the new ureteral stent, is the one that obtains the best results, showing minimum adverse effects and needing less ureteral manipulation

Estudio experimental comparativo de nuevos tratamientos endourológicos de la uropatía obstructiva mediante stents metálicos recubiertos

Tesis doctoral con la Mención de "Doctor Internacional"El manejo de la obstrucción ureteral de origen maligno o de carácter benigno complicado continúa siendo a día de hoy un desafío para la comunidad urológica. La disposición de catéteres ureterales doble jota como alternativa de derivación interna y de la nefrostomía percutánea como derivación externa, son las medidas terapéuticas que con mayor frecuencia se emplean en este tipo de pacientes. Sin embargo, la morbilidad asociada a ambas derivaciones provoca que sean necesarias otras vías de tratamiento. En este sentido, el empleo de stents metálicos podría satisfacer las expectativas, ya que permiten en un solo acto la resolución de la estenosis, sin derivaciones externas, y no se asocian con ninguno de los efectos adversos de las derivaciones urinarias actuales, y tampoco necesitan recambiar los catéteres ni la sonda de nefrostomía, lo que disminuye la calidad de vida de sus portadores. Sin embargo, a pesar de estas buenas perspectivas de éxito, cimentadas en los buenos resultados sobre todo de los stents vasculares, los stents ureterales no cumplen las perspectivas. La migración, la incrustación y sobre todo el crecimiento hiperplásico urotelial son las principales causas de que estos dispositivos no sean plenamente aceptados. El presente trabajo experimental en modelo porcino pretende evaluar la Hipótesis de que la alteración de la interacción entre los extremos del stent metálico, la pared ureteral y el peristaltismo ureteral disminuye la aparición de la reacción hiperplásica urotelial en los extremos del stent, lo que puede favorecer la expansión de los stents en el tracto urinario superior. Para ello se distribuyen los 60 animales del estudio en cinco grupos diferentes todos ellos con un modelo de estenosis ureteral experimental. Lo que permite evaluar diferentes técnicas diseñadas para disminuir o inhibir la formación de hiperplasia urotelial tras la disposición de un stent metálico. La primera técnica a evaluar es la realización de una endoureteromía en ambos extremos del stent, para de este modo alterar el peristaltismo en la zona de confluencia entre el stent y el uréter sano. Otro posibilidad terapéutica evaluada es la disposición de un novedoso diseño de stent ureteral híbrido y dinámico que no afecta al peristaltismo del uréter adyacente al stent y por ende, no debe producir hiperplasia. Por último, se evalúa en este estudio si la disposición coadyuvante de un catéter ureteral doble jota junto a un stent metálico ureteral disminuye la formación de hiperplasia urotelial. Los resultados del estudio experimental avalan la hipótesis de trabajo. Los grupos de estudio frente a sus respectivos grupos control evidencian que, la endoureterotomía en los extremos del stent disminuye significativamente el desarrollo de hiperplasia. Asimismo el nuevo diseño de stent metálico ureteral, HMS, es seguro, y efectivo en el tratamiento de la estenosis ureteral disminuyendo la formación de hiperplasia urotelial sin la necesidad de manipulación secundaria para asegurar la permeabilidad del stent. Por último, la disposición de un catéter ureteral coaxial al stent no disminuye la aparición de hiperplasia, pero sí su agresividad significativamente. Por todo ello, podemos concluir que la formación de hiperplasia urotelial relacionada con la disposición de stents ureterales metálicos disminuye significativamente cuando se reduce la interacción entre los extremos del stent y el ureter peristáltico adyacente. Además de que los catéteres doble jota no inhiben la formación de hiperplasia al asociarlos con los stents ureterales metálicos. De las tres terapias evaluadas, el nuevo stent ureteral, HMS, es el que mejores resultados alcanza, mínimos efectos adversos muestra y es el que menor manipulación ureteral provoca.The management of a malignant or complicated benign ureteral obstruction still today represents a challenge for the urological community. The placement of double J ureteral catheters as an alternative to internal diversion, and percutaneous nephrostomy as an external diversion, are the most frequently used therapeutic measures for this type of patient. However, morbidity associated with both diversions makes other treatment options necessary. In this sense, the use of metal stents could meet the expectations, as they allow stenosis to be solved in a single act, with no external diversions. In addition, they are not associated with any of the adverse effects caused by present urinary diversions and their catheters or nephrostomy catheter do not need to be replaced, which reduces carriers' quality of life. However, in spite of successful prospects, based on the positive results especially shown by vascular stents, ureteral stents do not meet our expectations. Migration, encrustation and especially urothelial hyperplasic growth are the main reasons why these devices are not wholly accepted. The present experimental study on swine model aims to assess the hypothesis that alteration in the interaction of the metal stent ends, ureteral wall and ureteral peristalsis reduces the onset of urothelial hyperplasic reaction at stent ends. This can favor stent expansion in the upper urinary tract. The 60 study animals are divided into five different groups, all of them having an experimental ureteral stenosis model. In this way, the different techniques designed to reduce or inhibit urothelial hyperplasia formation after metal stent placement are assessed. The first technique to be assessed is the performance of an endoureterotomy at both stent ends, so that peristalsis can be altered in the convergence area between stent and healthy ureter. Another therapeutic possibility assessed is to place an innovative design for a hybrid and dynamic ureteral stent, which does not involve the ureter peristalsis adjacent to the stent, and therefore should not cause hyperplasia. Finally, this study assesses whether coadjuvant placement of a double J ureteral catheter, together with a ureteral metal stent, reduces urothelial hyperplasia formation. Experimental study results support the working hypothesis. Study groups compared to their respective control groups prove that endoureterotomy at stent ends significantly reduces hyperplasia development. Likewise HMS, the new ureteral metal stent design, is safe and effective for the treatment of ureteral stenosis, as it decreases urothelial hyperplasia formation with no need for a secondary manipulation in order to ensure stent permeability. Finally, placing a ureteral catheter coaxially to the stent does not reduce hyperplasia onset. However, it does significantly reduce its aggressivity. Therefore, we can state that urothelial hyperplasia formation related to metal ureteral stent placement significantly decreases when interaction between stent ends and adjacent peristaltic ureter is reduced. In addition, double J catheters do not inhibit hyperplasia formation when they are associated with metal ureteral stents. Among the three therapies assessed, HMS, the new ureteral stent, is the one that obtains the best results, showing minimum adverse effects and needing less ureteral manipulation

Micro-ureteroscopy vs. ureteroscopy: effects of miniaturization on renal vascularization and intrapelvic pressure

Purpose Ureteroscopy (URS) is related to complications, as fever or postoperative urinary sepsis, due to high intrapelvic pressure (IPP) during the procedure. Micro-ureteroscopy (m-URS) aims to reduce morbidity by miniaturizing the instrument. The objective of this study is to compare IPP and changes in renal haemodynamics, while performing m-URS vs. conventional URS. Methods A porcine model involving 14 female pigs was used in this experimental study. Two surgeons performed 7 URS (8/9.8 Fr), for 45 min, and 7 m-URS (4.85 Fr), for 60 min, representing a total of 28 procedures in 14 animals. A catheter pressure transducer measured IPP every 5 min. Haemodynamic parameters were evaluated by Doppler ultrasound. The volume of irrigation fluid employed in each procedure was also measured. Results The range of average pressures was 5.08–14.1 mmHg in the m-URS group and 6.08–20.64 mmHg in the URS (NS). 30 mmHg of IPP were not reached in 90% of renal units examined with m-URS, as compared to 65% of renal units in the URS group. Mean peak diastolic velocity decreased from 15.93 to 15.22 cm/s (NS) in the URS group and from 19.26 to 12.87 cm/s in the m-URS group (p < 0.01). Mean resistive index increased in both groups (p < 0.01). Irrigation fluid volume used was 485 mL in the m-URS group and 1475 mL in the URS group (p < 0.001). Conclusions m-URS requires less saline irrigation volumes than the conventional ureteroscopy and increases renal IPP to a lesser extent.Authors received research funds from a public research institute (ISABIAL-FISABIO) and from Presurgy SL

Laparoscopic Partial Nephrectomy With Potassium-titanyl-phosphate Laser Versus Conventional Laparoscopic Partial Nephrectomy: An Animal Randomized Controlled Trial

OBJECTIVE To explore the feasibility, safety, and short-term results of potassium-titanyl-phosphate (KTP) laser laparoscopic partial nephrectomy (KTP-LPN) vs conventional laparoscopic partial nephrectomy (C-LPN). MATERIALS AND METHODS Thirty large white female pigs were randomized to KTP-LPN or C-LPN. Laparoscopic radical right nephrectomy was performed, and an artificial renal tumor was placed in the left kidney in 3 locations. A week later, 15 pigs underwent C-LPN and 15 underwent KTP-LPN. All C-LPNs were performed with renal ischemia. A 120-W setting was used, without arterial clamping in the KTPLPN group. Follow-up was done at day 1, week 3, and week 6. Retrograde pyelography was performed at 6 weeks, followed by animal sacrifice and necropsy. RESULTS All KTP-LPNs were performed without hilar clamping. C-LPNs were performed with hilar clamping, closing of the collecting system, and renorraphy. In the KTP laser group, 2 pigs died due to urinary fistula in the first week after surgery. In the C-LPN group, 1 pig died due to myocardial infarction and another due to malignant hyperthermia. Hemoglobin and hematocrit recovery were lower at 6 weeks in the KTP-LPN group. Renal function 24 hours after surgery was worse in the KTP-LPN group but recovered at 3 weeks and 6 weeks. No differences were observed in surgical margins. The necropsy showed no differences. Limitations of the study are the impossibility to analyze the collecting tissue sealing by the KTP, and the potential renal toxicity of the KTP laser. CONCLUSION Although KTP-LPN is feasible and safe in the animal model, further studies are needed. (C) 2016 Elsevier In

Molecular diagnosis of patients with epilepsy and developmental delay using a customized panel of epilepsy genes.

Pediatric epilepsies are a group of disorders with a broad phenotypic spectrum that are associated with great genetic heterogeneity, thus making sequential single-gene testing an impractical basis for diagnostic strategy. The advent of next-generation sequencing has increased the success rate of epilepsy diagnosis, and targeted resequencing using genetic panels is the a most cost-effective choice. We report the results found in a group of 87 patients with epilepsy and developmental delay using targeted next generation sequencing (custom-designed Haloplex panel). Using this gene panel, we were able to identify disease-causing variants in 17 out of 87 (19.5%) analyzed patients, all found in known epilepsy-associated genes (KCNQ2, CDKL5, STXBP1, SCN1A, PCDH19, POLG, SLC2A1, ARX, ALG13, CHD2, SYNGAP1, and GRIN1). Twelve of 18 variants arose de novo and 6 were novel. The highest yield was found in patients with onset in the first years of life, especially in patients classified as having early-onset epileptic encephalopathy. Knowledge of the underlying genetic cause provides essential information on prognosis and could be used to avoid unnecessary studies, which may result in a greater diagnostic cost-effectiveness

Mutations identified by a multigenic panel of epilepsy.

<p>Mutations identified by a multigenic panel of epilepsy.</p