Specificities and redundancies in the NEL family of bacterial E3 ubiquitin ligases of Salmonella enterica serovar Typhimurium.

Salmonella enterica serovar Typhimurium expresses two type III secretion systems, T3SS1 and T3SS2, which are encoded in Salmonella pathogenicity island 1 (SPI1) and SPI2, respectively. These are essential virulent factors that secrete more than 40 effectors that are translocated into host animal cells. This study focuses on three of these effectors, SlrP, SspH1, and SspH2, which are members of the NEL family of E3 ubiquitin ligases. We compared their expression, regulation, and translocation patterns, their role in cell invasion and intracellular proliferation, their ability to interact and ubiquitinate specific host partners, and their effect on cytokine secretion. We found that transcription of the three genes encoding these effectors depends on the virulence regulator PhoP. Although the three effectors have the potential to be secreted through T3SS1 and T3SS2, the secretion of SspH1 and SspH2 is largely restricted to T3SS2 due to their expression pattern. We detected a role for these effectors in proliferation inside fibroblasts that is masked by redundancy. The generation of chimeric proteins allowed us to demonstrate that the N-terminal part of these proteins, containing the leucine-rich repeat motifs, confers specificity towards ubiquitination targets. Furthermore, the polyubiquitination patterns generated were different for each effector, with Lys48 linkages being predominant for SspH1 and SspH2. Finally, our experiments support an anti-inflammatory role for SspH1 and SspH2.

Costos directos y calidad de vida de los pacientes con glaucoma en Paraguay / Direct costs and quality of life of patients with glaucoma in Paraguay

Introducción:El glaucoma constituyeun problema de salud pública por su alta prevalencia, los costos directos y la calidad de vida deteriorada de los pacientes que lo sufre. Objetivo:Determinar los costos directos y calidad de vida de los pacientes adultos con glaucoma atendidos en la Clínica Central de la Fundación Visión de noviembre 2022 a febrero 2023. Material y métodos:estudio observacional, descriptivo, corte transversal que incluyó pacientes con diagnóstico de glaucoma por muestreo por conveniencia. Resultados:Se incluyeron 140 pacientes entre 29 y 90 años, 52,9% femenino, 43,6% primaria incompleta, 32,1% desempleado. El tratamiento médico se realizó en el 81%, laser en 42% y procedimiento quirúrgico en 32%. Se informó un gasto promedio anual de 3.639.800gs ($ 503.93) por paciente. El estado emocional afectado se informó en el 35,7% de los pacientes, y calidad de vida deteriorada en el 48,6%; el 55% de los pacientes tiene dependencia para sus cuidados personales. Conclusión:El glaucoma implica grandes costos directos debido al tratamiento, además los pacientes tienen deterioro de la calidad de vida.Palabras Clave:glaucoma; costos de la atención en salud; distrés psicológico; calidad de vida

Introduction.Glaucoma is still a public health problem due to its high prevalence, the socioeconomic impact it entails for the patient who suffers from it, which decreases the quality of life. Objective:To determine the direct costs and quality of life of adult patients treated WITH glaucoma in the Fundación Visión Central Clinic from November 2022 to February 2023. Material and methods:observational, descriptive, cross-sectional study that included patients diagnosed with glaucoma by sampling by convenience. Results:140 patients between 29 and 90 years old were included, 52.9% female, 43.6% incomplete primary education, 32.1% unemployed. Medical treatment was performed in 81%, laser in 42% and surgical procedure in 32%. An average annual expense of 3,639,800gs ($503.93) per patient was reported. Emotional affectation was reported in 35.7% of the patients, and 43.5% deteriorated quality of life; 55% of the patients has dependency on personal care. Conclusion:Glaucoma involves large direct costs, due to treatment, in addition patients have deterioration in the quality of life.Key words:glaucoma; health care costs;psychological distress; quality of life

SNRPD2 Is a Novel Substrate for the Ubiquitin Ligase Activity of the Salmonella Type III Secretion Effector SlrP.

SlrP is a protein with E3 ubiquitin ligase activity that is translocated by Salmonella enterica serovar Typhimurium into eukaryotic host cells through a type III secretion system. A yeast two-hybrid screen was performed to find new human partners for this protein. Among the interacting proteins identified by this screen was SNRPD2, a core component of the spliceosome. In vitro ubiquitination assays demonstrated that SNRPD2 is a substrate for the catalytic activity of SlrP, but not for other members of the NEL family of E3 ubiquitin ligases, SspH1 and SspH2. The lysine residues modified by this activity were identified by mass spectrometry. The identification of a new ubiquitination target for SlrP is a relevant contribution to the understanding of the role of this Salmonella effector.

The NEL Family of Bacterial E3 Ubiquitin Ligases.

Some pathogenic or symbiotic Gram-negative bacteria can manipulate the ubiquitination system of the eukaryotic host cell using a variety of strategies. Members of the genera Salmonella, Shigella, Sinorhizobium, and Ralstonia, among others, express E3 ubiquitin ligases that belong to the NEL family. These bacteria use type III secretion systems to translocate these proteins into host cells, where they will find their targets. In this review, we first introduce type III secretion systems and the ubiquitination process and consider the various ways bacteria use to alter the ubiquitin ligation machinery. We then focus on the members of the NEL family, their expression, translocation, and subcellular localization in the host cell, and we review what is known about the structure of these proteins, their function in virulence or symbiosis, and their specific targets.

Experiencia con el uso de toxina botulínica en niños con vejiga hiperactiva neurogénica: Una serie de casos / Experience with Botulinum Toxin in Children with Neurogenic Overactive Bladder: A Case Series

Objetivo Describir la experiencia con la administración de inyección de toxina botulínica en niños con vejiga hiperactiva neurogénica refractaria a manejo de primera línea. Materiales y Métodos Estudio observacional descriptivo (serie de casos) que incluyó a 14 niños con diagnóstico de vejiga hiperactiva neurogénica refractarios a tratamiento de primera línea sometidos a administración intravesical de toxina botulínica entre 2015 y 2021; se realizó el seguimiento teniendo en cuenta las variables clínicas, con evaluación de la respuesta y de los eventos adversos. Se reportaron frecuencias absolutas y porcentajes para las variables cualitativas; para las variables cuantitativas, se reportaron medidas de tendencia central y dispersión. Resultados Se incluyeron 6 niños y 8 niñas, con una media de edad 10,1 (desviación estándar [DE]: ± 4,4) años. Todos los pacientes fueron tratados previamente con anticolinérgico y cateterismos limpios intermitentes, con una media de uso de 2,8 (DE: ± 1.0) pañales al día; 11 (78,5%) pacientes tenían antecedente de infección urinaria, 13 (92,8%), estreñimiento, y 2 (15,3%), incontinencia fecal. En la ecografía, 7 (50,0%) pacientes presentaban engrosamiento de las paredes vesicales, y 6 (42,8%), hidronefrosis. Tras el procedimiento, 1 paciente presentó infección urinaria como complicación, 6 presentaron una respuesta completa, 7, respuesta parcial, y 1 paciente no obtuvo respuesta con la primera inyección, con un tiempo promedio efectivo de la terapia 8 (DE: ± 6,3) meses. Una segunda inyección fue necesaria en 6 (42.8%) pacientes, y, de estos, 3 (50%) requirieron una tercera inyección. Conclusión La inyección de toxina botulinica intravesical como terapia de segunda línea de manejo para vejiga hiperactiva neurogénica tiene buenos resultados, con bajas tasas de complicaciones.

Objective To describe the experience with the administration of botulinum toxin injection in children with neurogenic overactive bladder who were refractory to the first-line management. Materials and Methods A descriptive observational study (case series) which included 14 children with a diagnosis of neurogenic overactive bladder who were refractory to the first-line treatment and were aubmitted to the intravesical administration of botulinum toxin between 2015 and 2021. Follow-up was performed taking into account the clinical variables, wth an evaluation of the response and the adverse events. Absolute frequencies and percentages were reported for the qualitative variables; for the quantitative variables, measures of central tendency and dispersion were reported. Results We included 6 boys and 8 girls with a mean age of 10.1 (standard deviation [SD]: ± 4.4) years. All patients were previously treated with anticholinergics and clean intermittent catheterizations, with a mean use of 2.8 (SD: ± 1.0) diapers per day; 11 (78.5%) had a history of urinary tract infection, 13 (92.8%), constipation, and 2 (15.3%), fecal incontinence. On ultrasound, 7 (50.0%) patients presented bladder wall thickening, and 6 (42.8%), hydronephrosis. After the procedure, 1 patient presented urinary tract infection as a complication, 6 presented complete response, 7, partial response, and 1 patient did not obtain a response with the first injection, with an effective mean time of therapy of 8 (SD: ± 6.3) months. A second injection was required by 6 (42.8%) patients, and of these, 3 (50%) required a third injection. Conclusion Intravesical botulinum toxin injection as a second-line management therapy for neurogenic overactive bladder yileds good results, with low rates of complications.

Non-productive angiogenesis disassembles Aß plaque-associated blood vessels.

The human Alzheimer's disease (AD) brain accumulates angiogenic markers but paradoxically, the cerebral microvasculature is reduced around Aß plaques. Here we demonstrate that angiogenesis is started near Aß plaques in both AD mouse models and human AD samples. However, endothelial cells express the molecular signature of non-productive angiogenesis (NPA) and accumulate, around Aß plaques, a tip cell marker and IB4 reactive vascular anomalies with reduced NOTCH activity. Notably, NPA induction by endothelial loss of presenilin, whose mutations cause familial AD and which activity has been shown to decrease with age, produced a similar vascular phenotype in the absence of Aß pathology. We also show that Aß plaque-associated NPA locally disassembles blood vessels, leaving behind vascular scars, and that microglial phagocytosis contributes to the local loss of endothelial cells. These results define the role of NPA and microglia in local blood vessel disassembly and highlight the vascular component of presenilin loss of function in AD.

A selective thioxothiazolidin-coumarin probe for Hg2+ based on its desulfurization reaction. Exploring its potential for live cell imaging.

Sensing the most toxic heavy metal (mercury) has attracted a lot of attention in recent years due to its extreme harmfulness to both human health and the environment. Thus, we reported herein the synthesis, spectroscopic and kinetic characterization, and biological evaluation of a new thioxothiazolidin coumarin derivative (ILA92), which undergoes a desulfurization reaction induced by mercuric ions (Hg2+). This process is the origin of a selective sensing of Hg2+ ions in aqueous solution by colorimetric and fluorescent methods. Furthermore, the probe showed great potential for imaging Hg2+ in living cells.

¿Qué factores se asocian con la reestrechez uretral en el paciente llevado a uretroplastia con o sin injerto? / Which Factors are Associated with Urethral re-Stricture in Patient Submitted to Urethroplasty with or without Grafting?

Objetivo Determinar los factores asociados a la reestrechez uretral en pacientes llevados a uretroplastia. Métodos Estudio observacional analítico retrospectivo. Se revisaron las historias clínicas de pacientes llevados a uretroplastia en el HSJ entre 2012-2017. Se realizó un análisis bivariado para estudiar las asociaciones entre la reestrechez uretral posterior a la uretroplastia y las caracteristicas preoperatorias, intraoperatorias y posoperatorias, así como un modelo de regresión logística múltiple. Resultados El total de pacientes llevados a uretroplastia fue de 82. La mediana de edad fue de 52 (RIQ = 35 -67) años. De todos los pacientes incluidos en el estudio 29 (36,2%) presentaron reestrechez. No se encontraron diferencias estadísticamente significativas en cuanto a los factores de riesgo en relación con reestrechez uretral en la población de estudio. Dentro de las técnicas quirúrgicas realizadas, los tipos de injerto utilizados en nuestros pacientes fueron la técnica con injerto de mucosa oral y con injerto de prepucio. De los pacientes a los cuales se les realizó uretroplastia con injerto de prepucio 13 (52%), presentaron reestrechez mostrando una diferencia estadísticamente significativa (p = 0,02). El tiempo quirúrgico fue mayor en los pacientes con reestrechez uretral (180 min RIQ = 120,0-189,0) comparado con los pacientes sin reestrechez (140 min RIQ = 120,0-198,0). En cuanto al modelo de regresión logística, pacientes llevados a uretroplastia con injerto de prepucio poseen más riesgo de presentar reestrechez uretral comparado con los pacientes a los cuales se les realizó la uretroplastia con otro tipo de injerto o con una tecnica terminoterminal sin injerto OR = 2,91 IC 95% (1,02-8,28). Conclusiones En nuestro estudio solo se encontró que el tipo de injerto utilizado es el único factor asociado al desarrollo de reestrechez uretral posterior a la realización de uretroplastia, otras variables clínicas descritas en la literatura no tuvieron asociación en nuestro estudio.

Objective To determine the association between risk factors, surgical procedure, and postoperative outcomes with stricture recurrence in patients who underwent urethroplasty. Methods An analytical observational study with retrospective data collection was conducted. We reviewed the medical records of all the patients who underwent urethroplasty at HSJ between the years 2012-2017. The analysis of the data included a bivariate model to study the association between the risk factors, surgical procedure, postoperative outcomes and stricture recurrence. Then, we used logistic regression analysis. Results 82 patients underwent urethroplasty: 29 (36.2%) presented stricture recurrence. The median age was 52 (IQR = 35-67) years old. There were no statistically significant differences between the risk factors, postoperative outcomes and stricture recurrence. In surgical technique we used buccal mucosal graft and penile skin graft. Stricture recurrence was observed in 13 patients who underwent penile skin graft urethroplasty showing a significant statistical difference (p = 0.02). The patients with stricture recurrence had longer operative time (180 minutes, IQR = 120.0-189.0) compared to patients without (140 minutes, IQR = 120-198). The logistic regression analysis showed that patients who underwent penile skin graft urethroplasty had a greater risk of stricture recurrence OR = 2,91 IC 95% (1,02-8,28). Conclusions The type of graft used is the only factor associated with the development of stricture recurrence after performing urethroplasty. Other clinical variables described in the literature have no association with our study.

Incidencia de hidronefrosis transitoria en reimplante vesicouretral abierto con técnica de Lich Gregoir en niños con reflujo vesicoureteral / Incidence of Transient Hydroneprosis in Open Vesicouretral Reimplant with Lich Gregoir Technique in Children with Vesicoureteral Reflux

Introducción y Objetivo La técnica de Lich Gregoir es actualmente la de elección para la corrección del reflujo vesicoureteral (RVU) de alto grado, con complicaciones como la obstrucción ureteral. La literatura reporta tasas del 6% al 28% de hidronefrosis transitoria, las cuales, en su gran mayoría, corresponden a un hallazgo temporal sin repercusión clínica. El objetivo de este trabajo es establecer la incidencia de hidronefrosis transitoria y la asociación con factores de riesgo. Materiales y Metodos Realizamos una cohorte descriptiva retrospectiva, analizando 119 unidades ureterales reimplantadas mediante técnica de Lich Gregoir (LG) en menores de 18 años, entre Septiembre del 2009 y abril del 2017. Se excluyeron pacientes con megauréter primario, RVU secundario y cirugía previa para corrección de RVU. El análisis estadístico se realizó con Stata 13.0 Resultados Evaluamos 85 pacientes, 34 de ellos reimplantados de manera bilateral, para un total de 119 unidades ureterales. El promedio de edad fue de 4,4 años y el 68% fueron mujeres. El RVU grado IV fue el más prevalente con el 42%. Las ecografías renales prequirúrgicas fueron normales en el 63% de los casos y el 14,2% presentó hidronefrosis transitoria con resolución a los 3 meses. No se reportaron casos de obstrucción ureteral. El grado de reflujo prequirúrgico no se asoció con hidronefrosis transitoria (p = 0,64) así como tampoco la lateralidad (p = 0,06). Conclusión El reimplante vesicoureteral abierto con técnica de Lich Gregoir en nuestra institución, reporta tasas de hidronefrosis transitoria comparables a la literatura mundial. No se identificaron factores de asociación para hidronefrosis transitoria, diferentes a la técnica quirúrgica perse.

Introduction and Objective Lich Gregoir technique is currently the choice for correction of vesicoureteral reflux (VUR), with complications such as ureteral obstruction. Literature reports rates from 6% to 28% of transient hydronephrosis, from which, most correspond to a temporary finding without clinical repercussion. This study aims to establish the incidence of transient hydronephrosis and the association with risk factors. Material and Methods In a descriptive retrospective cohort, we analyzed 119 ureteral units reimplanted with Lich Gregoir, under 18 years old, between September 2009 and April 2017. We have excluded patients with primary megaureter, VUR secondary and previous surgery for VUR correction. Statistical analysis was performed with Stata 13.0. Results We have evaluated 85 patients, 34 bilaterally reimplanted, totaling 119 ureteral units. Mean age was 4.4 years and 68% were women. VUR grade IV was the most prevalent with 42%. Pre-surgical renal ultrasounds were normal in 63% of the cases, and 14.2% had transient hydronephrosis with resolution at 3 months. No case of ureteral obstruction was reported. Pre-surgical reflux grade was not associated with transient hydronephrosis (p = 0.64) as well as neither laterality (p = 0.06). Conclusion Open Vesicoureteral reimplantaion with Lich Gregoir technique in our institution reports transient hydronephrosis rates similar to the literature. We have not found association factors with transient hydronephrosis, except for the surgical technique perse.

Hemorrhagic cystitis after hematopoietic stem cell transplantation: A challenge for the pediatric urologist.

OBJECTIVE: Hemorrhagic cystitis (HC) is a serious event that can occur after hematopoietic stem cell transplantation (HSCT). Treatment goals are primarily to preserve life, and then the functionality of the bladder. There is no standard therapeutic approach for HC. Described treatment options provide low success rates and are related to potential life-threatening side effects. The aim of this study was to describe our experience in treatment of HC following HSCT. PATIENTS AND METHODS: This was a retrospective study of patients with HC treated at our institution between January 2010 and October 2016. We analyzed demographics, underlying diagnosis, and treatment modalities. RESULTS: We treated 39 patients with HC. Mean age was 9.4 years (SD 4.20) and 64% were males. Acute leukemia was the most common underlying diagnosis in 27 (69%). Mean time from HSCT to HC onset was 55.46 days (SD 112.35). HC grades were: I (3), II (21), III (8), and IV (7). BK-viuria was present in 34 patients (87.2%). Non-invasive treatment was performed in 28 patients (71.8%). The remaining 11 (28.2%) required urological intervention (all high-grade), consisting of bladder irrigation in all of these. Additional treatments consisted of: intravesical cidofovir (4), intravesical sodium hyaluronate (5), cystoscopy and clot evacuation (4), selective angioembolization (2), percutaneous nephrostomy (1), and open extraction of bladder clots and cutaneous cystotomy (1). Overall, eight patients (20.5%) died as a result of the malignancy (3 in the urological intervention group), and of these four had active HC at death. Mean follow-up was 36.2 months (SD 24.9). CONCLUSION: HC is associated with high morbidity and mortality. Treatment should be individualized and designed to prioritize survival. However, bladder function should be preserved for the future.

Prevalencia de cicatrices retinocoroideas por toxoplasma en una población del Paraguay / Prevalence of toxoplasma retinochoroidal scars in a paraguayan population

El objetivo del trabajo fue determinar la prevalencia de cicatrices retinianas por toxoplasmosis en la población adulta en el municipio de San Francisco de Atyra en Paraguay. Se examinó dicha población desde julio del 2013 hasta marzo 2014, con la participaron en forma voluntaria de 61 familias seleccionadas al azar y dentro de cada una de ellas un integrante mayor de 18 años igualmente seleccionado al azar. Se realizó la toma de la agudeza visual utilizando la cartilla de Snellen en cada ojo por separado a 6 metros del paciente, refracción automatizada, prueba refractiva, biomicroscopía en lámpara de hendidura con énfasis en la profundidad de la cámara anterior de forma inmediata se descartaron pacientes con cámaras anteriores pandas o estrechas (utilización de gonioscopio mirrow 4 mini), presión intraocular, instilación de Tropicamida 0,5%/Fenilefrina HCL 5% en cada ojo, evaluación del polo posterior en lámpara de hendidura con lente de 90D Superfield, oftalmoscopia indirecta con lente de 20D y fotografía en la cámara no midriática. El diagnóstico de toxoplasmosis ocular fue basado en criterios clínicos, y aspectos de la cicatriz de la lesión por retinocoroiditis. Recibieron evaluación oftalmológica 61 pacientes de los cuales 6 pacientes fueron excluidos. La prevalencia de cicatrices corioretininanas fue del 9,1%, siendo más frecuente en hombres sin predilección de edad ni de lateralidad ocular, con agudeza visual igual o mejor a 20/40 con su mejor corrección.

The objective of this study was to determine the prevalence of retinal scars caused bytoxoplasmosis in an adult population from the municipality of San Francisco de Atyra inParaguay. The population of this town was examined from July, 2013 to March 2014.Sixty families, randomly selected, participated voluntarily and within each of them amember older than 18 years of age also selected randomly. Visual acuity was measuredusing the Snellen chart in each eye separately at 6 meters from the patient, as well as theautomated refraction, refraction test, biomicroscopy with a slit lamp with emphasis in thedepth of the anterior chamber. Patients with panda or narrow front chambers (use of amini 4 mirror gonioscope) were discarded. Intraocular pressure, instillation of tropicamide0.5%/phenylephrine HCl 5% in each eye, evaluation of the posterior pole in a slit lampwith 90D Superfield lens, indirect ophtalmoscopy with 20D lenses and photography with non-mydriatic camera were also carried out. The diagnosis of ocular toxoplasmosis wasbased in clinical criteria and aspects of the lesion scar by retinochoroiditis. Sixty onepatients were evaluated ophtalmologically and six were excluded. The prevalence ofchorioretinal scars was 9.1% being more frequent in men without age predominance orocular laterality, with visual acuity equal to or better than 20/40 with its best correction.

Cistitis quística glandular presentada como cistopatía quística / Cystitis cystica glandularis presented as cystic cystopathy

Se presenta el caso de una mujer de 73 años, con historia de hematuria macroscópica de aparición reciente, con presencia de quistes en toda la extensión de la mucosa vesical, y con diagnóstico por biopsias de vejiga, de una cistitis glandular. Se habla de cistopatía quística cuando los quistes se encuentran difusamente en la vejiga, y frecuentemente coexiste con lipomatosis pélvica en pacientes obesas, como la que reportada aquí. Es considerada una lesión premaligna y por eso es importante un seguimiento cuidadoso con cistoscopias periódicas. La cistitis quística se presenta más frecuentemente en hombres y en personas de mayor edad. Macroscópicamente, se manifiesta con nódulos submucosos únicos o múltiples, y los hallazgos histológicos son la presencia de nidos uroteliales redondeados, con dilatación quística dentro de la lámina propia, y paralelos a la superficie urotelial.

The case of a 73-year-old woman with a history of recent appearance of gross hematuria and diffuse cystic bladder lesions, diagnosed through a bladder biopsy with cystitis cystica glandularis is here in reported. Cystic cystopathy is reviewed. It is caractherized by cysts found diffusely throughout the bladder and it frequently coexists with pelvic lipomatosis in obese patients, such as the one in the case reported. It is considered a premalignant lesion, therefore careful follow-up with periodic cystoscopy is indicated. Cystitis cystica is found more often among males and elders. Macroscopically, it occurs as single or multiple submucosal nodules and the histological findings are the presence of rounded urothelial nests with cystic dilation within the lamina propria and parallel to the urothelial surface.