Revisión del género Cantabrogeus Salgado, 2000, con la descripción de tres nuevas especies hipogeas endémicas de la Cordillera Cantábrica (Coleoptera, Leiodidae, Cholevinae, Leptodirini)

Se revisan las dos especies previamente descritas del género Cantabrogeus (Coleoptera, Leiodidae, Cholevinae, Leptodirini), y se describen tres especies nuevas: Cantabrogeus antimachus sp. n., C. cultellus sp. n. y C. fresnedai sp. n. Una de ellas (C. antimachus sp. n.) es triploide (3n = 33) y la primera especie partenogenética conocida de Leiodidae. Se analizan y se discuten las relaciones existentes entre los diferentes taxones a partir de los caracteres morfológicos externos y las estructuras de los aparatos genitales masculino y femenino, así como las áreas cársticas que colonizan las diferentes especies en la zona cantábrica (NO ibérico). El análisis de datos moleculares (fragmentos de siete genes mitocondriales y nucleares) confirma la monofilia de Cantabrogeus y su proximidad filogenética con el género Fresnedaella, así como su inclusión dentro de la serie Quaestus, que es parafilética. La especie partenogenética (C. antimachus sp. n.) ocupa una posición derivada dentro del género Cantabrogeus, que, de acuerdo con estimas de la tasa de variación molecular basadas en estudios previos, probablemente se diversificó durante el Plioceno–Pleistoceno en las zonas cársticas cántabras, que son de una alta complejidad tectónica.The two previously known species of the genus Cantabrogeus (Coleoptera, Leiodidae, Cholevinae, Leptodirini) are redescribed, and three new species are described: Cantabrogeus antimachus n. sp., C. cultellus n. sp. and C. fresnedai n. sp. One of these (C. antimachus n. sp.) is tripliod (3n = 33) and the first known parthenogenetic species of Leiodidae. The relationships between the different taxa are discussed based on external morphological characters and the structure of male and female genitalia, as well as the distribution of the species in the different areas in the Cantabrian mountains (NW Iberia). Analyses of molecular data (fragments of seven mitochondrial and nuclear genes) confirms the monophyly of Cantabrogeus and its sister–group relationship with Fresnedaella, as well as its inclusion within the paraphyletic series Quaestus. The parthenogenetic species (C. antimachus n. sp.) has a derived position within Cantabrogeus, which, according to previous estimates of the molecular rate, probably diversified during the Pliocene–Pleistocene in the Cantabrian karst areas, of a high tectonic complexity.Se revisan las dos especies previamente descritas del género Cantabrogeus (Coleoptera, Leiodidae, Cholevinae, Leptodirini), y se describen tres especies nuevas: Cantabrogeus antimachus sp. n., C. cultellus sp. n. y C. fresnedai sp. n. Una de ellas (C. antimachus sp. n.) es triploide (3n = 33) y la primera especie partenogenética conocida de Leiodidae. Se analizan y se discuten las relaciones existentes entre los diferentes taxones a partir de los caracteres morfológicos externos y las estructuras de los aparatos genitales masculino y femenino, así como las áreas cársticas que colonizan las diferentes especies en la zona cantábrica (NO ibérico). El análisis de datos moleculares (fragmentos de siete genes mitocondriales y nucleares) confirma la monofilia de Cantabrogeus y su proximidad filogenética con el género Fresnedaella, así como su inclusión dentro de la serie Quaestus, que es parafilética. La especie partenogenética (C. antimachus sp. n.) ocupa una posición derivada dentro del género Cantabrogeus, que, de acuerdo con estimas de la tasa de variación molecular basadas en estudios previos, probablemente se diversificó durante el Plioceno–Pleistoceno en las zonas cársticas cántabras, que son de una alta complejidad tectónica

Somatic embryogenesis and efficient regeneration of Vitis vinifera L. 'Carménère' plants

Research Note

Inflammatory cell death induced by cytotoxic lymphocytes: a dangerous but necessary liaison

Cytotoxic lymphocytes (CLs), and more specifically Tc and NK cells, are the main executors of cell death in the immune system, playing a key role during both immunosurveillance and immunotherapy. These cells induce regulated cell death (RCD) by different mechanisms, being granular exocytosis and expression of death ligands the most prominent and best characterized ones. Apoptosis, a traditionally considered low-inflammatory type of cell death, has been accepted for years as the paradigm of RCD induced by CLs. However, several recent studies have demonstrated that NK cells and Tc cells can also induce more inflammatory forms of cell death, namely, necroptosis, pyroptosis, and ferroptosis. Activation of these highly inflammatory types of cell death appears to critically contribute to the activation of a successful antitumour immune response. Additionally, the role of specific cell death pathways in immunogenic cell death is still under intense debate, especially considering the interconnections with other inflammatory forms of cell death. These evidences, together with the advent of new cancer immunotherapies, highlight the necessity to deepen our understanding of the link between the cell death triggered by CLs and inflammation. This knowledge will be instrumental to maximize the antitumour potential of immunotherapies, minimizing deleterious effects associated with these treatments. In this review, we will briefly summarize the main features of apoptosis, necroptosis, pyroptosis and ferroptosis, to subsequently discuss the most recent evidences about the role of these RCD pathways during the elimination of cancer cells mediated by CLs and its modulation to increase the efficacy of cancer immunotherapy

The Multifaceted Function of Granzymes in Sepsis: Some Facts and a Lot to Discover

Sepsis is a serious global health problem. In addition to a high incidence, this syndrome has a high mortality and is responsible for huge health expenditure. The pathophysiology of sepsis is very complex and it is not well-understood yet. However, it is widely accepted that the initial phase of sepsis is characterized by a hyperinflammatory response while the late phase is characterized by immunosuppression and immune anergy, increasing the risk of secondary infections. Granzymes (Gzms) are a family of serine proteases classified according to their cleavage specificity. Traditionally, it was assumed that all Gzms acted as cytotoxic proteases. However, recent evidence suggests that GzmB is the one with the greatest cytotoxic capacity, while the cytotoxicity of others such as GzmA and GzmK is not clear. Recent studies have found that GzmA, GzmB, GzmK, and GzmM act as pro-inflammatory mediators. Specially, solid evidences show that GzmA and GzmK function as extracellular proteases that regulate the inflammatory response irrespectively of its ability to induce cell death. Indeed, studies in animal models indicate that GzmA is involved in the cytokine release syndrome characteristic of sepsis. Moreover, the GZM family also could regulate other biological processes involved in sepsis pathophysiology like the coagulation cascade, platelet function, endothelial barrier permeability, and, in addition, could be involved in the immunosuppressive stage of sepsis. In this review, we provide a comprehensive overview on the contribution of these novel functions of Gzms to sepsis and the new therapeutic opportunities emerging from targeting these proteases for the treatment of this serious health problem

Optimization of molecular detection of GD2 synthase mRNA in retinoblastoma

Extraocular dissemination is the main cause of death in patients with retinoblastoma in developing countries and there are few molecular markers that could be used for evaluation of minimal disseminated disease. The expression of the ganglioside GD2 is present in retinoblastoma cells metastatic to the bone marrow and the enzyme GD2 synthase activity is detected in neuroblastoma, which shares many phenotypic features with retinoblastoma. Our purpose was to optimize the detection of GD2 synthase expression by reverse transcription-polymerase chain reaction (RT-PCR) followed by nested-PCR in human retinoblastoma cell lines and patient samples. The optimization strategy was carried out by using the retinoblastoma cell lines Y79 and WERI-Rb1 and specific primers designed for the human sequence of the GD2 synthase mRNA. We detected GD2 synthase expression with at least 200 pg and 40 pg of total RNA extracted from cultured retinoblastoma cells, using a first round of RT-PCR amplification and a second round of nested-PCR, respectively. We have also confirmed the detection of GD2 synthase by RT-PCR and immunohistochemical expression of the ganglioside in human retinoblastoma tumors xenotransplanted in nude mice. In a study from tumor bank specimens from 8 retinoblastoma patients, we were able to demonstrate the presence of GD2 synthase mRNA in blood and cerebrospinal fluid samples in cases of extraocular dissemination of the tumor. The sequence was not detected in samples from children with low-risk disease or healthy adult volunteers. The detection of GD2 synthase mRNA through an optimized nested RT-PCR assay may be a promising tool for the assessment of minimal disseminated disease in enucleated patients.Fil: Laurent, Viviana Eunice. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Servicio de Hemato-Oncología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; ArgentinaFil: Otero, Laura L.. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; ArgentinaFil: Vazquez, Valeria. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; ArgentinaFil: Camarero, Sandra. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; ArgentinaFil: Gabri, Mariano Rolando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; ArgentinaFil: Labrada, Maria. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; ArgentinaFil: Garcia de Davila, Maria Teresa. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan"; ArgentinaFil: Chantada, Guillermo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital de Pediatría "Juan P. Garrahan". Servicio de Hemato-Oncología; ArgentinaFil: Alonso, Daniel Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Oncología Molecular; Argentin

Tumor urotelial de vejiga en la infancia: reporte de un nuevo caso

Introduction: bladder tumors are rare in childhood, only 0.38% of cases belong to people younger than 20 years prevailing the mesodermic origin. Considering those of epithelial origin, more than 100 cases have been diagnosed since 1950, mostly with superficial lesions, benign behavior, more common in males (ratio male / female 5/1 to 7/1), and its etiology has not yet been clarified. Case presentation: it is reported the case of a 15-year-old black teenager who came to our hospital with a total painless haematuria with no other associated symptoms. After some studies like transabdominal ultrasound and cystoscopy with biopsy it was found that he had a malignant tumor of the vesical urothelium. Discussion: papillomatous vesical tumor in the right lateral side of the bladder, away from the urethral meatus with approximately 2 cm of diameter and narrow base similar to bladder mucosa coloration. Partial cystectomy was performed evolving without complications. Conclusions: urothelial tumors in the pediatric population are extremely rare, its biological behavior is less aggressive than the one in the adult population, and the election treatment is the Trans urethral resection.Introducción: los tumores vesicales son infrecuentes en la infancia, solo un 0,38% de los casos corresponden a menores de 20 años, prevaleciendo los de origen mesodérmico. Considerando aquellos de estirpe epitelial, algo más de 100 casos han sido diagnosticados desde 1950, tratándose, en su gran mayoría, de lesiones superficiales, de comportamiento benigno, más frecuentes en varones (cociente hombre/mujer de 5/1 a 7/1), su etiología aún no ha sido aclarada.Presentación de caso: se reporta el caso de un adolescente de 15 años de edad raza negra que acude a nuestro hospital por hematuria total indolora, sin otros síntomas asociados. Después de los estudios realizados como Ecografía transabdominal y Cistoscopia con biopsia se comprueba que presentaba una tumoración maligna del urotelio vesical.Discusión:tumoración vesical de aspecto papilomatoso, en la cara lateral derecha de la vejiga alejada del meato ureteral, de aproximadamente 2 cm de diámetro, de base estrecha y de coloración similar a la mucosa de vejiga. Se le realizo Cistectomía parcial evolucionando sin complicaciones.Conclusiones: los tumores uroteliales en la población pediátrica, son extremadamente raros, su comportamiento biológico es menos agresivo que el que tienen en la población adulta, el tratamiento de elección es la Resección Tran­suretral

Granzyme A inhibition reduces inflammation and increases survival during abdominal sepsis

Aims: Peritonitis is one of the most common causes of sepsis, a serious syndrome characterized by a dysregulated systemic inflammatory response. Recent evidence suggests that Granzyme A (GzmA), a serine protease mainly expressed by NK and T cells, could act as a proinflammatory mediator and could play an important role in the pathogenesis of sepsis. This work aims to analyze the role and the therapeutic potential of GzmA in the pathogenesis of peritoneal sepsis. Methods: The level of extracellular GzmA as well as GzmA activity were analyzed in serum from healthy volunteers and patients with confirmed peritonitis and were correlated with the Sequential Organ Failure Assessment (SOFA) score. Peritonitis was induced in C57Bl/6 (WT) and GzmA-/- mice by cecal ligation and puncture (CLP). Mice were treated intraperitoneally with antibiotics alone or in combination serpinb6b, a specific GzmA inhibitor, for 5 days. Mouse survival was monitored during 14 days, levels of some proinflammatory cytokines were measured in serum and bacterial load and diversity was analyzed in blood and spleen at different times. Results: Clinically, elevated GzmA was observed in serum from patients with abdominal sepsis suggesting that GzmA plays an important role in this pathology. In the CLP model GzmA deficient mice, or WT mice treated with an extracellular GzmA inhibitor, showed increased survival, which correlated with a reduction in proinflammatory markers in both serum and peritoneal lavage fluid. GzmA deficiency did not influence bacterial load in blood and spleen and GzmA did not affect bacterial replication in macrophages in vitro, indicating that GzmA has no role in bacterial control. Analysis of GzmA in lymphoid cells following CLP showed that it was mainly expressed by NK cells. Mechanistically, we found that extracellular active GzmA acts as a proinflammatory mediator in macrophages by inducing the TLR4-dependent expression of IL-6 and TNFa. Conclusions: Our findings implicate GzmA as a key regulator of the inflammatory response during abdominal sepsis and provide solid evidences about its therapeutic potential for the treatment of this severe pathology

Factores de riesgo en pacientes con cáncer de riñón

Introduction: renal adenocarcinoma described by Koing and known as Grawitz tumor represents approximately 3% of all malignant neoplasias in human beings and the female – male ratio of 2.1 is more frequent in the urban area and the most affected patients are between 50 and 70 years internationally. Objective: to identify the influence of certain risk factors in the onset of the disease. Method: it was performed a retro descriptive and prospective study where it was analyzed the surgical treatment of kidney cancer in the University Province Hospital "Carlos Manuel de Céspedes", since January 2008 to January 2013, distributing the patients according to age and sex, symptoms, treatment, metastasis and survival time. Results: it was found that the cancer was more frequent in women between 50 and 59 years. The prevailing symptoms were lumbar pain and hematuria. The most frequent metastases were in the liver and in soft parts. The deceased were 4 patients: 3 women and a man. Conclusions: the mostly used therapeutic procedure was the radical nephrectomy; the urinary infection prevailed as a complication. The clear cell carcinoma was the histological type that prevailed. Survival was greater in males. The right renal tumor prevailed on the left tumor.Introducción: el adenocarcinoma renal descrito por Koing y conocido como tumor de Grawitz representa aproximadamente el 3 % de todas las neoplasias malignas en el ser humano y la proporción masculina femenina  de 2.1 resulta más frecuente en la zona urbana y la mayoría de los pacientes afectados se encuentran entre los 50 y 70 años a nivel internacional.Objetivo: identificar la influencia de determinados factores de riesgo en la aparición de esta enfermedad.Método: se realizó un estudio descriptivo y retroprospectivo donde se analizó el tratamiento quirúrgico del cáncer de riñón en el Hospital Provincial Universitario “Carlos Manuel de Céspedes”, desde Enero de 2008 a Enero 2013, distribuyendo los pacientes por edad y sexo, síntomas encontrados, tratamiento realizado, metástasis y tiempo de sobrevida. Resultados: se determinó que dicho cáncer fue más frecuente en el sexo femenino  entre 50 y 59 años. El síntoma predomínate fue el dolor lumbar y hematuria. Las metástasis más frecuentes fueron en Hígado y partes blandas. Los fallecidos fueron de 4 pacientes, 3 mujeres y un hombre.Conclusiones: el proceder terapéutico utilizado con mayor frecuencia fue la nefrectomía radical; la infección urinaria predomino como complicación. Resultó el carcinoma de células claras el tipo histológico que prevaleció. La supervivencia  fue mayor en el sexo masculino. El tumor renal derecho predomino sobre el izquierdo