Lats1 and Lats2 are required for the maintenance of multipotency in the Müllerian duct mesenchyme.

WNT signaling plays essential roles in the development and function of the female reproductive tract. Although crosstalk with the Hippo pathway is a key regulator of WNT signaling, whether Hippo itself plays a role in female reproductive biology remains largely unknown. Here, we show that conditional deletion of the key Hippo kinases Lats1 and Lats2 in mouse Müllerian duct mesenchyme cells caused them to adopt the myofibroblast cell fate, resulting in profound reproductive tract developmental defects and sterility. Myofibroblast differentiation was attributed to increased YAP and TAZ expression (but not to altered WNT signaling), leading to the direct transcriptional upregulation of Ctgf and the activation of the myofibroblast genetic program. Müllerian duct mesenchyme cells also became myofibroblasts in male mutant embryos, which impeded the development of the male reproductive tract and resulted in cryptorchidism. The inactivation of Lats1/2 in differentiated uterine stromal cells in vitro did not compromise their ability to decidualize, suggesting that Hippo is dispensable during implantation. We conclude that Hippo signaling is required to suppress the myofibroblast genetic program and maintain multipotency in Müllerian mesenchyme cells.

The granulosa cell response to luteinizing hormone is partly mediated by YAP1-dependent induction of amphiregulin.

BACKGROUND: The LH surge is a pivotal event that triggers multiple key ovarian processes including oocyte maturation, cumulus expansion, follicular wall rupture and luteinization of mural granulosa and theca cells. Recently, LH-dependent activation of the Hippo signaling pathway has been shown to be required for the differentiation of granulosa cells into luteal cells. Still, the precise interactions between Hippo and LH signaling in murine granulosa cells remain to be elucidated. METHODS: To detect the expression of effectors of the Hippo pathway, western blot, immunohistochemical and RT-qPCR analyses were performed on granulosa cells treated with LH in vitro or isolated from immature mice treated with eCG and hCG. Cultured granulosa cells were pretreated with pharmacologic inhibitors to identify the signaling pathways involved in Hippo regulation by LH. To study the roles of Yap1 and Taz in the regulation of the LH signaling cascade, RT-qPCR and microarray analyses were done on granulosa cells from Yap1f/f;Tazf/f mice treated with an adenovirus to drive cre expression. RT-qPCR was performed to evaluate YAP1 binding to the Areg promoter following chromatin immunoprecipitation of granulosa cells collected from mice prior to or 60 min following hCG treatment. RESULTS: Granulosa cells showed a transient increase in LATS1, YAP1 and TAZ phosphorylation levels in response to the ovulatory signal. This Hippo activation by LH was mediated by protein kinase A. Furthermore, Yap1 and Taz are required for the induction of several LH target genes such as Areg, Pgr and Ptgs2, and for the activation of the ERK1/2 pathway. Consistent with these results, there was a substantial overlap between genes that are upregulated by LH and those that are downregulated following loss of Yap1/Taz, highlighting a major role for Hippo in mediating LH actions in the ovulation process. Finally, we showed that there is a marked recruitment of YAP1 to the Areg promoter of granulosa cells in response to hCG stimulation. CONCLUSIONS: Overall, these results indicate that Hippo collaborates with the cAMP/PKA and ERK1/2 pathways to participate in the precise regulation of the LH cascade, and that Areg, as a direct transcriptional target of YAP1, is involved in mediating its actions in the ovary. Video Abstract.

Lats1 and Lats2 are required for ovarian granulosa cell fate maintenance.

Recent reports suggest that the Hippo signaling pathway influences ovarian follicle development; however, its exact roles remain unknown. Here, we examined the ovarian functions of the Hippo kinases large tumor suppressors (LATS)1 and 2, which serve to inactivate the transcriptional coactivators Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ). Inactivation of Lats1/2 in murine granulosa cells either in vitro or in vivo resulted in a loss of granulosa cell morphology, function, and gene expression. Mutant cells further underwent changes in structure and gene expression suggestive of epithelial-to-mesenchymal transition and transdifferentiation into multiple lineages. In vivo, granulosa cell-specific loss of Lats1/2 caused the ovarian parenchyma to be mostly replaced by bone tissue and seminiferous tubule-like structures. Transdifferentiation into Sertoli-like cells and osteoblasts was attributed in part to the increased recruitment of YAP and TAZ to the promoters of sex-determining region Y box 9 and bone γ-carboxyglutamate protein, key mediators of male sex determination and osteogenesis, respectively. Together, these results demonstrate for the first time a critical role for Lats1/2 in the maintenance of the granulosa cell genetic program and further highlight the remarkable plasticity of granulosa cells.-Tsoi, M., Morin, M., Rico, C., Johnson, R. L., Paquet, M., Gévry, N., Boerboom, D. Lats1 and Lats2 are required for ovarian granulosa cell fate maintenance.

The DNA double-strand "breakome" of mouse spermatids.

De novo germline mutations arise preferentially in male owing to fundamental differences between spermatogenesis and oogenesis. Post-meiotic chromatin remodeling in spermatids results in the elimination of most of the nucleosomal supercoiling and is characterized by transient DNA fragmentation. Using three alternative methods, DNA from sorted populations of mouse spermatids was used to confirm that double-strand breaks (DSB) are created in elongating spermatids and repaired at later steps. Specific capture of DSB was used for whole-genome mapping of DSB hotspots (breakome) for each population of differentiating spermatids. Hotspots are observed preferentially within introns and repeated sequences hence are more prevalent in the Y chromosome. When hotspots arise within genes, those involved in neurodevelopmental pathways become preferentially targeted reaching a high level of significance. Given the non-templated DNA repair in haploid spermatids, transient DSBs formation may, therefore, represent an important component of the male mutation bias and the etiology of neurological disorders, adding to the genetic variation provided by meiosis.

Novel exons in the tbx5 gene locus generate protein isoforms with distinct expression domains and function.

TBX5 is the gene mutated in Holt-Oram syndrome, an autosomal dominant disorder with complex heart and limb deformities. Its protein product is a member of the T-box family of transcription factors and an evolutionarily conserved dosage-sensitive regulator of heart and limb development. Understanding TBX5 regulation is therefore of paramount importance. Here we uncover the existence of novel exons and provide evidence that TBX5 activity may be extensively regulated through alternative splicing to produce protein isoforms with differing N- and C-terminal domains. These isoforms are also present in human heart, indicative of an evolutionarily conserved regulatory mechanism. The newly identified isoforms have different transcriptional properties and can antagonize TBX5a target gene activation. Droplet Digital PCR as well as immunohistochemistry with isoform-specific antibodies reveal differential as well as overlapping expression domains. In particular, we find that the predominant isoform in skeletal myoblasts is Tbx5c, and we show that it is dramatically up-regulated in differentiating myotubes and is essential for myotube formation. Mechanistically, TBX5c antagonizes TBX5a activation of pro-proliferative signals such as IGF-1, FGF-10, and BMP4. The results provide new insight into Tbx5 regulation and function that will further our understanding of its role in health and disease. The finding of new exons in the Tbx5 locus may also be relevant to mutational screening especially in the 30% of Holt-Oram syndrome patients with no mutations in the known TBX5a exons.

Endovascular treatment of cerebral sinus thrombosis due to vaccine-induced immune thrombotic thrombocytopenia.

INTRODUCTION: There is little data on the role of endovascular treatment (EVT) of cerebral venous sinus thrombosis (CVST) due to vaccine-induced immune thrombotic thrombocytopenia (VITT). Here, we describe clinical characteristics and outcomes of CVST-VITT patients who were treated with EVT. PATIENTS AND METHODS: We report data from an international registry of patients who developed CVST within 28 days of SARS-CoV-2 vaccination, reported between 29 March 2021 and 6 March 2023. VITT was defined according to the Pavord criteria. RESULTS: EVT was performed in 18/136 (13%) patients with CVST-VITT (92% aspiration and/or stent retrieval, 8% local thrombolysis). Most common indications were extensive thrombosis and clinical or radiological deterioration. Compared to non-EVT patients, those receiving EVT had a higher median thrombus load (4.5 vs 3). Following EVT, local blood flow was improved in 83% (10/12, 95% confidence interval [CI] 54-96). One (6%) asymptomatic sinus perforation occurred. Eight (44%) patients treated with EVT also underwent decompressive surgery. Mortality was 50% (9/18, 95% CI 29-71) and 88% (8/9, 95% CI 25-66) of surviving EVT patients achieved functional independence with a modified Rankin Scale score of 0-2 at follow-up. In multivariable analysis, EVT was not associated with increased mortality (adjusted odds ratio, 0.66, 95% CI 0.16-2.58). DISCUSSION AND CONCLUSION: We describe the largest cohort of CVST-VITT patients receiving EVT. Half of the patients receiving EVT died during hospital admission, but most survivors achieved functional independence.

KiT-GENIE, the French genetic biobank of kidney transplantation.

KiT-GENIE is a monocentric DNA biobank set up to consolidate the very rich and homogeneous DIVAT French cohort of kidney donors and recipients (D/R) in order to explore the molecular factors involved in kidney transplantation outcomes. We collected DNA samples for kidney transplantations performed in Nantes, and we leveraged GWAS genotyping data for securing high-quality genetic data with deep SNP and HLA annotations through imputations and for inferring D/R genetic ancestry. Overall, the biobank included 4217 individuals (n = 1945 D + 2,272 R, including 1969 D/R pairs), 7.4 M SNPs and over 200 clinical variables. KiT-GENIE represents an accurate snapshot of kidney transplantation clinical practice in Nantes between 2002 and 2018, with an enrichment in living kidney donors (17%) and recipients with focal segmental glomerulosclerosis (4%). Recipients were predominantly male (63%), of European ancestry (93%), with a mean age of 51yo and 86% experienced their first graft over the study period. D/R pairs were 93% from European ancestry, and 95% pairs exhibited at least one HLA allelic mismatch. The mean follow-up time was 6.7 years with a hindsight up to 25 years. Recipients experienced biopsy-proven rejection and graft loss for 16.6% and 21.3%, respectively. KiT-GENIE constitutes one of the largest kidney transplantation genetic cohorts worldwide to date. It includes homogeneous high-quality clinical and genetic data for donors and recipients, hence offering a unique opportunity to investigate immunogenetic and genetic factors, as well as donor-recipient interactions and mismatches involved in rejection, graft survival, primary disease recurrence and other comorbidities.

[Use of the insertable Holter with remote detection in the etiological diagnosis of cryptogenic stroke: Analysis of 73 patients]. / Utilidad del Holter insertable con detección remota en el diagnóstico etiológico del ictus criptogénico: análisis de 73 pacientes.

INTRODUCTION: Cryptogenic stroke constitutes 25% of all ischemic strokes, of which 20-30% are due to atrial fibrillation (AF). With the aim of increasing the detection rate, implantable long-term monitoring devices have emerged. The study of the profile of the ideal candidate subsidiary to such monitoring would provide a better understanding of the mechanisms underlying this subtype of stroke. OBJECTIVE: To determine which variables are related and can predict the detection of silent AF in patients with cryptogenic stroke. PATIENTS AND METHODS: This is a longitudinal cohort with recruitment from March 2017 to May 2022. They are patients with an implantable monitoring device and cryptogenic stroke with a minimum monitoring of one year. RESULTS: The total number of patients included was 73, with a mean age of 58.8 years, 56.2% were male. AF was detected in 21 patients (28.8%). The most frequent cardiovascular risk factors were hypertension (47.9%) and dyslipidemia (45.2%). The most frequent topography was cortical (52%). Regarding the echocardiographic parameters, 22% had a dilated left atrium, 19% had a patent foramen ovale, and 22% had high-density supraventricular tachycardia (>1%) on Holter monitoring. In the multivariate analysis, the only variable that predicts AF is the presence of high-density supraventricular tachycardia, with an area under the curve of 0.726 (CI 0.57-0.87, p=0.04), sensitivity of 47.6%, specificity of 97.5%, positive predictive value of 90.9%, negative predictive value of 78.8%, and accuracy of 80.9%. CONCLUSIONS: The presence of high-density supraventricular tachycardia can be indicative for predicting silent AF. No other variables have been observed that allow us to predict detection of AF in these patients.

Endoscopic retrograde cholangiopancreatography for treatment biliopleural fistulas.

BACKGROUND: Biliopleural fistula is a rare communication between the biliary system and the pleural space secondary to ruptured hydatid cysts, hepatobiliary surgeries or penetrating wounds. There is not so much of the subject in the literature, so there is no standardized protocol for its management. OBJECTIVE: The objective of this work is to determine advantages and disadvantages of the use of ERCP as a conservative initial treatment for biliopleural fistulas. METHOD: Our retrospective study included patients with a diagnosis of biliopleural fistula who underwent ERCP as initial treatment, older than 18 years in a period from August 2016 to August 2019. RESULTS: Eight patients with a diagnosis of biliopleural fistula were included, 75% men and 25% women with a mean age of 24.5 years; the traumatic etiology was predominant in our group (89%), the diagnosis was made between days 3 and 14 after chest tube placement and we were successful in treating the fistula with ERCP in 87.5% of our patients. CONCLUSIONS: The use of ERCP as initial treatment for biliopleural fistula should be considered as the initial tool in the algorithm of management of this entity.

ANTECEDENTES: La fístula biliopleural es una comunicación poco frecuente entre el sistema biliar y el espacio pleural secundario a ruptura de quistes hidatídicos, cirugías hepatobiliares o heridas penetrantes Existe poco sobre el tema en la literatura por lo que no existe un protocolo estandarizado para su manejo. OBJETIVO: El objetivo de este trabajo es determinar ventajas y desventajas del uso de CPRE como tratamiento inicial conservador para fístulas biliopleurales. MÉTODO: Nuestro estudio retrospectivo incluyó pacientes con diagnóstico de fístula biliopleural quienes fueron sometidos a CPRE como tratamiento inicial, mayores de 18 años en un período de Agosto de 2016 a Agosto de 2019. RESULTADOS: Se incluyeron 8 pacientes con diagnístico de fístula biliopleural, 75% hombres y 25% mujeres con una media de edad de 24.5 años; la etiología traumática fue a predominante en nuestro grupo (89%), el diagnóstico se realizó entre los días 3 y 14 post colocación de sonda torácica y obtuvimos éxito en el tratamiento de la fístula con CPRE en el 87.5% de nuestros pacientes. CONCLUSIONES: El uso de CPRE como tratamiento inicial para fístula biliopleural debe ser considerada como la herramienta inicial en el algoritmo de manejo de esta entidad.

Quiste de duplicación gástrica: inusual presentación en adultos, tratado con éxito por laparoscopía.

Las duplicaciones gastrointestinales son malformaciones congénitas raras que pueden presentarse en diferentes partes del tracto gastrointestinal, de tamaño y sintomatología variable. Las duplicaciones gástricas representan el 3-5% de estas malformaciones. La presentación más frecuente es la quística, en la mayoría de los casos en la curvatura mayor. El diagnóstico es raro en la edad adulta y el tratamiento de elección es quirúrgico, el cual puede llevarse a cabo de forma convencional o por mínima invasión. Se presenta el caso de una mujer adulta con diagnóstico de quiste de duplicación gástrica en fondo gástrico adyacente a la unión gastroesofágica a quien se realizó resección total por laparoscopia.Gastrointestinal duplications are rare congenital malformations, they can appear in different parts of the gastrointestinal tract, of variable size and symptomatology. Gastric duplications represent 3-5% of these malformations, the most frequent presentation is statistical, located mostly in the greater curvature, the diagnosis is rare in adulthood, the treatment of choice is surgical, which can be carried out in a conventional or minimally invasive manner. We present the case of an adult woman diagnosed with a gastric duplication cyst in the gastric fundus adjacent to the gastroesophageal junction, who underwent total resection by laparoscopy.

Trauma cervical penetrante: experiencia de un hospital de enseñanza en México / Penetrating cervical trauma: experience of a teaching hospital in Mexico

Introducción. El trauma cervical penetrante es una entidad poco frecuente, con tendencia al alza, de considerable morbimortalidad. Es fundamental conocer su manejo por cirujanos en entrenamiento, quienes en su mayoría son comúnmente los proveedores del tratamiento en las áreas de urgencias. El objetivo de este estudio fue demostrar la experiencia de un centro de trauma en México en el tratamiento de trauma cervical penetrante administrado por cirujanos en entrenamiento. Métodos. Estudio retrospectivo de pacientes con diagnóstico de trauma cervical penetrante sometidos a exploración cervical entre los años 2014 y 2019. Se identificaron 110 expedientes, se excluyeron 26 por falta de datos indispensables para la investigación y se analizaron los datos de 84 pacientes. Resultados. El 96,4 % de los pacientes fueron hombres, la mayoría entre 16 y 50 años (83,3 %) y la zona anatómica II fue la más comúnmente lesionada (65,4 %). Las etiologías más frecuentes fueron heridas por arma cortopunzante (67,9 %) y heridas por proyectil de arma de fuego (25 %). El 95 % de los procedimientos fueron realizados por residentes de cuarto y quinto año. La mediana de días de estancia hospitalaria fue de 2 días. La incidencia de complicaciones fue de 9,5 % y la mortalidad de 1,2 %. Conclusiones. Los cirujanos generales en etapa de formación entrenados en centros de trauma tienen la capacidad de tratar de forma óptima el trauma cervical penetrante sin modificación de las tasas de morbimortalidad descritas en la literatura internacional.

Introduction. Penetrating cervical trauma is a rare entity, with an upward trend, of considerable morbidity and mortality. It is essential to acknowledge its management by surgeons in training, who are usually the providers in the emergency areas. The objective of this study was to demonstrate the experience of a trauma center in Mexico in the treatment of penetrating cervical trauma by surgeons in training. Methods. This was a retrospective study of patients diagnosed with penetrating cervical trauma who underwent cervical examination between 2014 and 2019; 110 records were identified, 26 were excluded due to lack of essential data for the analysis, and the data of 84 patients were included. Results. 96.4% of the patients were men, the majority between 16 and 50 years old (83.3%) and the anatomic zone II was the most frequently injured (65.4%). The most common etiologies were stab wounds (67.9%) and gunshot wounds (25%). Ninety five percent of the procedures were performed by fourth- and fifth-year residents. The median number of days of hospital stay was 2 (2-4) days. The incidence of complications was 9.5% and mortality in 1.2%. Conclusions. General surgeons in the trauma training stage can optimally treat penetrating cervical trauma with the same morbidity and mortality rates described in the international literature.

[Women and strokes. Hospital epidemiology in Spain]. / El ictus en la mujer. Epidemiología hospitalaria en España.

INTRODUCTION: Although in clinical practice no distinction is usually made in the prevention and management of strokes according to the sex of the patient, some studies suggest the existence of significant clinical differences in the strokes suffered by females. PATIENTS AND METHODS: The purpose of our study was to confirm and characterize these differences in patients admitted to hospitals in our community; to do so, we performed an analysis of the stroke or little stroke patients included in the Spanish Neurology Society's Stroke Database (BADISEN), from 1996 to 2001, belonging to eight Spanish hospitals. We analyzed the social aspects, risk factors, clinical features, diagnostic data, situation and referral on discharge of 4,248 female patients (39%). A univariate and, later, a multivariate analysis were performed using logistic regression of the variables that showed significant differences (p<0.2). RESULTS: The findings from our study confirm a number of differences between male and female stroke victims. Strokes occur later in females (71 years old compared with 69 in males), when the patient lives alone. The following also occur more frequently in these patients than in males: arterial hypertension (62.8 versus 51.9%, p<0.0001), cardiac insufficiency (8.0 as opposed to 4.5%, p<0.0001) and atrial fibrillation (21.4 versus 11.3%, p<0.0001) as risk factors; cardioembolic causation (22.4 against 15.4%, p<0.0001) in anterior circulation (44.0 compared to 36.1%, p<0.0001); severity is increased (6.8 on the Canadian scale versus 7.5%, p<0.0001); more medical complications (40.5 compared to 33.7%, p<0.0001), more serious sequelae (64.2 on the Barthel index compared to 72.9%, p<0.0001) and a higher institutionalization rate, among other distinguishing facts. CONCLUSIONS: Such differences must be taken into account in the prevention and management of strokes in females.

[Effectiveness of lacosamide in the treatment of refractory neuropathic pain: an open observational trial]. / Efectividad de la lacosamida en el tratamiento del dolor neuropatico refractario: estudio observacional abierto.

INTRODUCTION: Although different treatments are available for neuropathic pain, these patients are often refractory, which makes it necessary to test treatments that, as they have proven useful in other pathologies, could be effective in neuropathic pain. PATIENTS AND METHODS: The study made use of the medical records of patients who had been treated with lacosamide for neuropathic pain in different hospitals in the central area of the peninsula and who fulfilled similar characteristics in terms of refractoriness to other standard treatments, in a follow-up that lasted at least six months, or who had had to stop treatment with that drug for some reason or another. A sample of 114 patients (61 males and 53 females) with a mean age of 60.5 years was obtained from the data. RESULTS: The most common causes of neuropathic pain were: diabetic polyneuropathy (31.6%), post-herpes neuralgia (22.8%), trigeminal neuralgia (17.5%), suboccipital and lumbar-radicular neuralgia (both 12.3%). Effectiveness was good/very good in most patients, with the mean score on the visual analogue scale after six months dropping from 7.7 to 4.8. No serious side effects were reported in any of the patients, but in 12 and 10 patients no recordings were made beyond six months, due to ineffectiveness and intolerance to the treatment, respectively. CONCLUSIONS: Treatment with lacosamide in neuropathic pain due to different causes could be considered an effective and well-tolerated alternative for patients who fail to respond to standard treatments.

TITLE: Efectividad de la lacosamida en el tratamiento del dolor neuropatico refractario: estudio observacional abierto.Introduccion. Aunque se dispone de diferentes tratamientos para el dolor neuropatico, en muchas ocasiones estos pacientes son refractarios, lo que hace necesario probar tratamientos que, por su utilidad en otras patologias, podrian ser eficaces en el dolor neuropatico. Pacientes y metodos. Se recogieron las historias clinicas de pacientes que hubieran sido tratados con lacosamida para el dolor neuropatico, en diferentes hospitales de la zona centro peninsular, y que cumplieran unas caracteristicas similares en cuanto a refractariedad a otros tratamientos estandares, en un seguimiento de al menos seis meses, o que hubiesen tenido que suspender el tratamiento con dicho farmaco por cualquier motivo. Se obtuvo una muestra de 114 pacientes, 61 varones y 53 mujeres, con una edad media de 60,5 años. Resultados. Las causas de dolor neuropatico mas frecuentes fueron: polineuropatia diabetica (31,6%), neuralgia postherpetica (22,8%), neuralgia del trigemino (17,5%), neuralgia suboccipital y lumbociatalgia (un 12,3% en ambas). La eficacia fue buena/muy buena en la mayoria de los pacientes, con un descenso medio en la escala analogica visual tras seis meses de 7,7 a 4,8. No se registraron efectos secundarios graves en ningun paciente, pero en 12 y 10 pacientes no hubo registro mas alla de seis meses, por ineficacia e intolerancia al tratamiento, respectivamente. Conclusiones. El tratamiento con lacosamida en el dolor neuropatico de diferentes causas podria considerarse como una alternativa efectiva y bien tolerada para aquellos pacientes que no respondan a los tratamientos estandares.

Distinct expression and function of alternatively spliced Tbx5 isoforms in cell growth and differentiation.

Mutations in the T-box transcription factor Tbx5 cause Holt-Oram syndrome, an autosomal dominant disease characterized by a wide spectrum of cardiac and upper limb defects with variable expressivity. Tbx5 haploinsufficiency has been suggested to be the underlying mechanism, and experimental models are consistent with a dosage-sensitive requirement for Tbx5 in heart development. Here, we report that Tbx5 levels are regulated through alternative splicing that generates, in addition to the known 518-amino-acid protein, a C-terminal truncated isoform. This shorter isoform retains the capacity to bind DNA, but its interaction with Tbx5 collaborators such as GATA-4 is altered. In vivo, the two spliced isoforms are oppositely regulated in a temporal and growth factor-dependent manner and are present in distinct DNA-binding complexes. The expression of the long isoform correlates with growth stimulation, and its reexpression in postnatal transgenic mouse hearts promotes hypertrophy. Conversely, the upregulation of the short but not the long isoform in C2C12 myoblasts leads to growth arrest and cell death. The results provide novel insight into posttranscriptional Tbx5 regulation and point to an important role not only in cell differentiation but also in cell proliferation and organ growth. The data may help analyze genotype-phenotype relations in patients with Holt-Oram syndrome.