Glaucoma avanzado. Guía de práctica clínica / Advanced glaucoma. Clinical practice guideline

objetivo Realización de una guía de práctica clínica actualizada que sirva de orientación para la detección, el manejo y el tratamiento de pacientes con glaucoma avanzado en la edad adulta. Métodos Tras la definición de los objetivos y alcance de la guía se constituyó el grupo de trabajo que formuló las preguntas clínicas estructuradas siguiendo el formato PICO (Patient, Intervention, Comparison, Outcomes). Se realizó una revisión de la literatura publicada hasta el momento, incluyendo guías de práctica clínica internacionales, utilizándose las herramientas AMSTAR-2 (Assessment of Multiple systematic Rewiews) y «Risk of Bias» de Cochrane para la evaluación de la calidad de la información de forma independiente por, al menos, 2 revisores. El nivel de evidencia y la elaboración del grado de recomendación se establecieron siguiendo la metodología del Scottish Intercollegiate Guidelines Network (SIGN). Resultados Se presentan recomendaciones con sus correspondientes niveles de evidencia que pueden ser de utilidad para la detección, el seguimiento y el tratamiento de pacientes con glaucoma avanzado en la edad adulta. Conclusiones A pesar de que la evidencia científica existente es escasa y el nivel de evidencia para muchas de las preguntas planteadas no es muy alto, esta guía de práctica clínica ofrece una revisión actualizada de las recomendaciones existentes para el manejo del glaucoma avanzado en el adulto (AU)

Objective To present an update clinical practice guideline that serve as a guide for the detection, evaluation and treatment of adults patients with advanced glaucoma. Methods After defining the objectives and scope of the guide, the working group was formed and structured clinical questions were formulated following the PICO (Patient, Intervention, Comparison, Outcomes) format. Once all the existing clinical evidence had been independently evaluated with the AMSTAR-2 (Assessment of Multiple systematic Rewiews) and Cochrane «Risk of bias» tools by at least 2 reviewers, recommendations were formulated following the Scottish Intercollegiate methodology Guideline Network (SIGN). Results Recommendations with their corresponding levels of evidence that may be useful in the diagnosis, monitoring and treatment of adults patients with advanced glaucoma. Conclusions Despite the fact that for many of the questions the level of scientific evidence available is not very high, this clinical practice guideline offers an updated review of the different existing aspects related to the evaluation and management of advanced glaucoma (AU)

Advanced glaucoma. Clinical practice guideline.

OBJECTIVE: To present an update clinical practice guideline that serve as a guide for the detection, evaluation and treatment of adults patients with advanced glaucoma. METHODS: After defining the objectives and scope of the guide, the working group was formed and structured clinical questions were formulated following the PICO (Patient, Intervention, Comparison, Outcomes) format. Once all the existing clinical evidence had been independently evaluated with the AMSTAR 2 (Assessment of Multiple systematic Rewiews) and Cochrane "Risk of bias" tools by at least two reviewers, recommendations were formulated following the Scottish Intercollegiate methodology. Guideline Network (SIGN). RESULTS: Recommendations with their corresponding levels of evidence that may be useful in the diagnosis, monitoring and treatment of adults patients with advanced glaucoma. CONCLUSIONS: Despite the fact that for many of the questions the level of scientific evidence available is not very high, this clinical practice guideline offers an updated review of the different existing aspects related to the evaluation and management of advanced glaucoma.

Multidisciplinary virtual management of pulmonary nodules.

INTRODUCTION AND OBJECTIVES: Multidisciplinary nodule clinics provide high-quality care and favor adherence to guidelines. Virtual care has shown savings benefits along with patient satisfaction. Our aim is to describe the first year of operation of a multidisciplinary virtual lung nodule clinic, the population evaluated and issued decisions. Secondarily, among discharged patients, we aimed to analyze their follow-up prior to the existence of our consultation, evaluating its adherence to guidelines. MATERIALS AND METHODS: Observational study including all patients evaluated at the Virtual Lung Nodule Clinic (VLNC) (March 2018- March 2019). Clinical and radiological data were recorded. Recommendations, based on 2017 Fleischner Society guidelines, were categorized into follow-up, discharge or referral to lung cancer consultation. Discharged patients were classified according to adherence to guidelines of their previous management, into adequate, prolonged and non-indicated follow-up. RESULTS: A total of 365 patients (58.9% men; median age 64.0 years) were included. Sixty-four percent had smoking history and 23% had chronic obstructive pulmonary disease (COPD). Most nodules were solid (87.4%) and multiple (57.5%). The median diameter was 6.00 mm. 43.8% of patients were discharged following first VLNC evaluation. Among them, 27.5% had received appropriate follow-up, but 66.9% had received poor management. Patients with prolonged follow-up (33.1%) were older (67.0 vs 60.5 years) and had larger nodules (6.00 mm vs 5.00). Non-indicated follow-up patients (33.8%) were more non-smokers (77.8% vs 31.8%) and presented smaller nodules (4.00 vs 5.00 mm). CONCLUSIONS: During its first year of operation, the VLNC has evaluated a population with a relevant risk profile for lung cancer development, management of which should be cautious and adhere to guidelines. After the first VLNC assessment, approximately one-half of this population was discharged. It was noticeable that previous follow-up of discharged patients was found poorly adherent to guidelines, with a marked tendency to overmanagement.

AMP-activated protein kinase (AMPK) signaling in GnRH neurons links energy status and reproduction.

BACKGROUND: Reproduction is tightly coupled to body energy and metabolic status. GnRH neurons, master elements and final output pathway for the brain control of reproduction, directly or indirectly receive and integrate multiple metabolic cues to regulate reproductive function. Yet, the molecular underpinnings of such phenomenon remain largely unfolded. AMP-activated protein kinase (AMPK), the fundamental cellular sensor that becomes activated in conditions of energy deficit, has been recently shown to participate in the control of Kiss1 neurons, essential gatekeepers of the reproductive axis, by driving an inhibitory valence in situations of energy scarcity at puberty. However, the contribution of AMPK signaling specifically in GnRH neurons to the metabolic control of reproduction remains unknown. METHODS: Double immunohistochemistry (IHC) was applied to evaluate expression of active (phosphorylated) AMPK in GnRH neurons and a novel mouse line, named GAMKO, with conditional ablation of the AMPK α1 subunit in GnRH neurons, was generated. GAMKO mice of both sexes were subjected to reproductive characterization, with attention to puberty and gonadotropic responses to kisspeptin and metabolic stress. RESULTS: A vast majority (>95%) of GnRH neurons co-expressed pAMPK. Female (but not male) GAMKO mice displayed earlier puberty onset and exaggerated LH (as surrogate marker of GnRH) responses to kisspeptin-10 at the prepubertal age. In adulthood, GAMKO females retained increased LH responsiveness to kisspeptin and showed partial resilience to the inhibitory effects of conditions of negative energy balance on the gonadotropic axis. The modulatory role of AMPK in GnRH neurons required preserved ovarian function, since the differences in LH pulsatility detected between GAMKO and control mice subjected to fasting were abolished in ovariectomized animals. CONCLUSIONS: Altogether, our data document a sex-biased, physiological role of AMPK signaling in GnRH neurons, as molecular conduit of the inhibitory actions of conditions of energy deficit on the female reproductive axis.

Kisspeptin treatment induces gonadotropic responses and rescues ovulation in a subset of preclinical models and women with polycystic ovary syndrome.

STUDY QUESTION: Can kisspeptin treatment induce gonadotrophin responses and ovulation in preclinical models and anovulatory women with polycystic ovary syndrome (PCOS)? SUMMARY ANSWER: Kisspeptin administration in some anovulatory preclinical models and women with PCOS can stimulate reproductive hormone secretion and ovulation, albeit with incomplete efficacy. WHAT IS KNOWN ALREADY: PCOS is a prevalent, heterogeneous endocrine disorder, characterized by ovulatory dysfunction, hyperandrogenism and deregulated gonadotrophin secretion, in need of improved therapeutic options. Kisspeptins (encoded by Kiss1) are master regulators of the reproductive axis, acting mainly at GnRH neurons, with kisspeptins being an essential drive for gonadotrophin-driven ovarian follicular maturation and ovulation. Altered Kiss1 expression has been found in rodent models of PCOS, although the eventual pathophysiological role of kisspeptins in PCOS remains unknown. STUDY DESIGN, SIZE, DURATION: Gonadotrophin and ovarian/ovulatory responses to kisspeptin-54 (KP-54) were evaluated in three preclinical models of PCOS, generated by androgen exposures at different developmental windows, and a pilot exploratory cohort of anovulatory women with PCOS. PARTICIPANTS/MATERIALS, SETTING, METHODS: Three models of PCOS were generated by exposure of female rats to androgens at different periods of development: PNA (prenatal androgenization; N = 20), NeNA (neonatal androgenization; N = 20) and PWA (post-weaning androgenization; N = 20). At adulthood (postnatal day 100), rats were subjected to daily treatments with a bolus of KP-54 (100 µg/kg, s.c.) or vehicle for 11 days (N = 10 per model and treatment). On Days 1, 4, 7 and 11, LH and FSH responses were assessed at different time-points within 4 h after KP-54 injection, while ovarian responses, in terms of follicular maturation and ovulation, were measured at the end of the treatment. In addition, hormonal (gonadotrophin, estrogen and inhibin B) and ovulatory responses to repeated KP-54 administration, at doses of 6.4-12.8 nmol/kg, s.c. bd for 21 days, were evaluated in a pilot cohort of anovulatory women (N = 12) diagnosed with PCOS, according to the Rotterdam criteria. MAIN RESULTS AND THE ROLE OF CHANCE: Deregulated reproductive indices were detected in all PCOS models: PNA, NeNA and PWA. Yet, anovulation was observed only in NeNA and PWA rats. However, while anovulatory NeNA rats displayed significant LH and FSH responses to KP-54 (P < 0.05), which rescued ovulation, PWA rats showed blunted LH secretion after repeated KP-54 injection and failed to ovulate. In women with PCOS, KP-54 resulted in a small rise in LH (P < 0.05), with an equivalent elevation in serum estradiol levels (P < 0.05). Two women showed growth of a dominant follicle with subsequent ovulation, one woman displayed follicle growth but not ovulation and desensitization was observed in another patient. No follicular response was detected in the other women. LIMITATIONS, REASONS FOR CAUTION: While three different preclinical PCOS models were used in order to capture the heterogeneity of clinical presentations of the syndrome, it must be noted that rat models recapitulate many but not all the features of this condition. Additionally, our pilot study was intended as proof of principle, and the number of participants is low, but the convergent findings in preclinical and clinical studies reinforce the validity of our conclusions. WIDER IMPLICATIONS OF THE FINDINGS: Our first-in-rodent and -human studies demonstrate that KP-54 administration in anovulatory preclinical models and women with PCOS can stimulate reproductive hormone secretion and ovulation, albeit with incomplete efficacy. As our rat models likely reflect the diversity of PCOS phenotypes, our results argue for the need of personalized management of anovulatory dysfunction in women with PCOS, some of whom may benefit from kisspeptin-based treatments. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by research agreements between Ferring Research Institute and the Universities of Cordoba and Edinburgh. K.S. was supported by the Wellcome Trust Scottish Translational Medicine and Therapeutics Initiative (STMTI). Some of this work was undertaken in the MRC Centre for Reproductive Health which is funded by the MRC Centre grant MR/N022556/1. M.T.-S. is a member of CIBER Fisiopatología de la Obesidad y Nutrición, which is an initiative of Instituto de Salud Carlos III. Dr Mannaerts is an employee of Ferring International PharmaScience Center (Copenhagen, Denmark), and Drs Qi, van Duin and Kohout are employees of the Ferring Research Institute (San Diego, USA). Dr Anderson and Dr Tena-Sempere were recipients of a grant support from the Ferring Research Institute, and Dr Anderson has undertaken consultancy work and received speaker fees outside this study from Merck, IBSA, Roche Diagnostics, NeRRe Therapeutics and Sojournix Inc. Dr Skorupskaite was supported by the Wellcome Trust through the Scottish Translational Medicine and Therapeutics Initiative 102419/Z/13/A. The other authors have no competing interest.

SIRT1 mediates obesity- and nutrient-dependent perturbation of pubertal timing by epigenetically controlling Kiss1 expression.

Puberty is regulated by epigenetic mechanisms and is highly sensitive to metabolic and nutritional cues. However, the epigenetic pathways mediating the effects of nutrition and obesity on pubertal timing are unknown. Here, we identify Sirtuin 1 (SIRT1), a fuel-sensing deacetylase, as a molecule that restrains female puberty via epigenetic repression of the puberty-activating gene, Kiss1. SIRT1 is expressed in hypothalamic Kiss1 neurons and suppresses Kiss1 expression. SIRT1 interacts with the Polycomb silencing complex to decrease Kiss1 promoter activity. As puberty approaches, SIRT1 is evicted from the Kiss1 promoter facilitating a repressive-to-permissive switch in chromatin landscape. Early-onset overnutrition accelerates these changes, enhances Kiss1 expression and advances puberty. In contrast, undernutrition raises SIRT1 levels, protracts Kiss1 repression and delays puberty. This delay is mimicked by central pharmacological activation of SIRT1 or SIRT1 overexpression, achieved via transgenesis or virogenetic targeting to the ARC. Our results identify SIRT1-mediated inhibition of Kiss1 as key epigenetic mechanism by which nutritional cues and obesity influence mammalian puberty.

Cisto gastro anastomosis laparoscópica / Laparoscopic cysto gastro anastomosis

Tras el Symposium de Atlanta (1992), existe acuerdo en definir un pseudoquiste pancreático (PQP) como una colección de jugo pancreático, rico en amilasa, rodeado por una pared no epitelial, producido como consecuencia de una pancreatitis aguda, pancreatitis crónica o traumatismo pancreático. Se estima que se necesitan al menos cuatro semanas para que constituya la pared definida que distingue un PQP de una colección líquida. Caso Clínico. Presentamos el caso de un paciente de 29 años de edad, con antecedentes de pancreatitis aguda de etiología biliar. Se realizó colecistectomía laparoscópica una vez resuelta la pancreatitis y egresó a domicilio. 40 días después refiere dolor abdominal, saciedad precoz progresiva hasta intolerancia a la vía oral, nausea y vómito, hechos que motivaron la reinternación. La tomografía computada revela un PQP gigante de 19.18 por 19.55 centímetros. Se plantea abordaje laparoscópico, realizándose cisto-gastroanastomosis con sutura mecánica por vía laparoscópica. CONCLUSIÓN: La cisto-gastroanastomosis laparoscópica es una opción terapéutica para el tratamiento del pseudoquiste pancreático porque ofrece: drenaje continuo, bajo índice de recidiva y pocas complicaciones que superan al tratamiento endoscópico y al drenaje guiado por imagenología, junto con las ventajas de mínima invasión.

After the Atlanta Symposium (1992), there's an agreement to define a pancreatic pseudocyst (PPC) as a collection of pancreatic juice, therefore rich in amylase, surrounded by a non-epithelial wall, produced as a consequence of an acute pancreatitis, chronic pancreatitis or pancreatic trauma. It is estimated that it takes at least four weeks for it to constitute the defined wall that distinguishes a PPQ from a fluid collection. Clinical case. We present the case of a 29-year-old patient with a history of acute pancreatitis of biliary etiology. Laparoscopic cholecystectomy was performed once the pancreatitis resolved and was outpatient. 40 days later he presents severe abdominal pain, early satiety and finally intolerance to the oral intake, nausea and vomiting, reasons for readmission. The computed tomography revealed a giant PPC of 19.18 by 19.55 centimeters which was resolved by laparoscopic cysto-gastrostomy with stapler. CONCLUSION: Laparoscopic cysto-gastrostomy is a therapeutic option for the treatment of pancreatic pseudocyst because it offers: continuous drainage, low rate of recurrence and few complications that overcome endoscopic treatment and drainage guided by imaging, with the advantages of minimal invasion.

Efectividad de la profilaxis antibiótica en pacientes con colecistitis aguda sometidos a colecistectomía laparoscópica / Effectiveness of antibiotic prophylaxis in acute cholecystitis patients undergoing laparoscopic cholecystectomy

OBJETIVO: Determinar la efectividad de la profilaxis antibiótica con Cefazolina en pacientes sometidos a colecistectomía laparoscópica por Colecistitis Aguda MATERIAL Y MÉTODOS: Cohorte Prospectiva POBLACIÓN: Pacientes mayores de 18 años, con patología litiásica vesicular aguda, sometidos a colecistectomía laparoscópica. SEDE Y TEMPORALIDAD: Hospital Obrero Nº 1 de la Caja Nacional de Salud La Paz ­ Bolivia. Período comprendido entre el 1 de Julio de 2016 al el 31 de Diciembre de 2016. RESULTADOS: Se incluyeron 95 pacientes con Colecistitis Aguda divididos en dos grupos, el Grupo A (SIN profilaxis antibiótica) compuesto por 50 sujetos y el Grupo B (CON profilaxis antibiótica) de 45 sujetos. La edad promedio fue de 48 años, el peso de 70 kilos, la talla de 165 cm y el IMC de 27,4 km/ m2. El tiempo operatorio promedio fue de 50 (±22,815) minutos en el total del grupo, 45 min. (±18,460) en el grupo A y 60 min (±24,862) en el grupo B. La conversión a cirugía abierta fue de 9 sujetos (9,5%). La infección del sitio operatorio se presentó en 47 sujetos (49,5%), 30 sujetos (60%) EN EL GRUPO A y 18 en el grupo B (40%). El OR calculado es de 0,444 (IC 95% 0,195 ­ 1,011). CONCLUSIONES: La administración de Cefazolina en forma profiláctica, parece no disminuir la probabilidad de infección del sitio operatorio en colecistitis aguda abordada por laparoscopía

OBJECTIVE: To determine the effectiveness of antibiotic prophylaxis with Cefazolin in patients undergoing laparoscopic cholecystectomy due to Acute Cholecystitis. METHODS: Prospective Cohort POPULATION: Adult patients (older than 18 years), with acute lithiasic cholecystitis, who underwent laparoscopic cholecystectomy. PLACE AND TEMPORALITY: Hospital Obrero No. 1 of the Caja Nacional de Salud La Paz ­ Bolivia, from July to December 2016. RESULTS: A total of 95 patients with Acute Cholecystitis were enrolled and divided in to two groups, group A (without antibiotic prophylaxis) composed of 50 subjects and Group B, (with antibiotic prophylaxis) 45 subjects. The mean age was 48 years old, weight 70 Kg, hight 165 cm and a BMI of 27.4 kg/M2. The mean operating time was 50 minutes (+- 22.185), group A 45 minutes and group B 60 min. Conversion to open surgery happened in 9 patients (9,5%), all in group B. Surgical Site infection (SSI) occurred in 47 patients (49.5%), of whom 30 patients belong to group A (60%) and 18 patients to group B (40%). The calculated Odds ratio is 0.444 (IC 95% 0,195-1.011). There were no bile duct injuries or morality in this study. CONCLUSIONS: The prophylactic administration of Cefazolin does not seems to decrease the probability of SSI in acute cholecystitis treated laparoscopically.

Reparación laparoscópica de hernia hiatal paraesofágica con material protésico / Laparoscopic repair of paraesophageal hiatal hernia with prosthetic material

El manejo quirúrgico de las Hernias Hiatales Paraesofágicas puede ser realizado por toracotomía izquierda o por laparoscopía. Ambos abordajes son aceptables, pero actualmente se ha incrementado mucho el abordaje laparoscópico por las características de mínima invasividad, pero con la utilización de material protésico como forma de refuerzo, cuando el hiato esofágico es superior a 5 cm. Los objetivos del abordaje laparoscópico son la reducción anatómica completa y la competencia del esfínter esofágico inferior. Estos objetivos se consiguen con el cierre de la crura diafragmática y la funduplicatura laparoscópica. Se presentan dos casos clínicos donde se ha utilizado malla de Poliester fijada con tackers colocada en forma de C con la apertura hacia la cara anterior y el solapamiento por delante del esófago. CONCLUSIÓN: El uso de material protésico en la reparación del defecto de las Hernias Hiatales Gigantes es una alternativa razonable.

The surgical management of Paraesophageal Hiatal Hernia can be performed by left thoracotomy or by laparoscopy. Both approaches are acceptable, but currently the laparoscopic approach has increased greatly due to the characteristics of minimal invasiveness, but with the use of prosthetic material as a form of reinforcement, when the esophageal hiatus is greater than 5 cm. The objectives of the laparoscopic approach are a complete anatomic reduction and lower esophageal sphincter competition. These objectives are achieved with the closure of the diaphragmatic crural and laparoscopic fundoplication. Two clinical cases were presented where Polyester mesh, fixed with tackers, was placed in a C shaped disposition with the opening facing the anterior and the overlap in front of the esophagus. CONCLUSION: The use of prosthetic material in the repair of the defect of Giant Hiatal Hernia is a reasonable alternative.

[Planar molecular arrangements aid the design of MHC class II binding peptides].

The coupling between peptides and MHC-II proteins in the human immune system is not well understood. This work presents an evidence-based hypothesis of a guiding intermolecular force present in every human MHC-II protein (HLA-II). Previously, we examined the spatial positions of the fully conserved residues in all HLA-II protein types. In each one, constant planar patterns were revealed. These molecular planes comprise of amino acid groups of the same chemical species (for example, Gly) distributed across the protein structure. Each amino acid plane has a unique direction and this directional element offers spatial selectivity. Constant within all planes, too, is the presence of an aromatic residue possessing electrons in movement, leading the authors to consider that the planes generate electromagnetic fields that could serve as an attractive force in a single direction. Selection and attraction between HLA-II molecules and antigen peptides would, therefore, be non-random, resulting in a coupling mechanism as effective and rapid as is clearly required in the immune response. On the basis of planar projections onto the HLA-II groove, modifications were made by substituting the key residues in the class II-associated invariant chain peptide-a peptide with a universal binding affinity-resulting in eight different modified peptides with affinities greater than that of the unmodified peptide. Accurate and reliable prediction of MHC class II-binding peptides may facilitate the design of universal vaccine-peptides with greatly enhanced binding affinities. The proposed mechanisms of selection, attraction and coupling between HLA-II and antigen peptides are explained further in the paper.

Testicular expression of the Lin28/let-7 system: Hormonal regulation and changes during postnatal maturation and after manipulations of puberty.

The Lin28/let-7 system, which includes the RNA-binding proteins, Lin28a/Lin28b, and let-7 miRNAs, has emerged as putative regulator of puberty and male gametogenesis; yet, its expression pattern and regulation in postnatal testis remain ill defined. We report herein expression profiles of Lin28 and let-7 members, and related mir-145 and mir-132, in rat testis during postnatal maturation and in models of altered puberty and hormonal deregulation. Neonatal expression of Lin28a and Lin28b was low and rose markedly during the infantile period; yet, expression patterns diverged thereafter, with persistently elevated levels only for Lin28b, which peaked at puberty. Let-7a, let-7b, mir-132 and mir-145 showed profiles opposite to Lin28b. In fact, let-7b and mir-145 were abundant in pachytene spermatocytes, but absent in elongating spermatids, where high expression of Lin28b was previously reported. Perturbation of puberty by neonatal estrogenization reverted the Lin28/let-7 expression ratio; expression changes were also detected in other models of delayed puberty, due to early photoperiod or nutritional manipulations. In addition, hypophysectomy or growth hormone (GH) deficiency revealed regulation of this system by gonadotropins and GH. Our data document the expression profiles of the Lin28/let-7 system in rat testis along postnatal/pubertal maturation, and their perturbation in models of pubertal and hormonal manipulation.

Effects and interactions of tachykinins and dynorphin on FSH and LH secretion in developing and adult rats.

Kisspeptin/neurokinin B/dynorphin (KNDy) neurons, which coexpress kisspeptins (Kps), neurokinin B (NKB), and dynorphin (Dyn), regulate gonadotropin secretion. The KNDy model proposes that NKB (a stimulator, through NK3R) and Dyn (an inhibitor, through κ-opioid receptor) shape Kp secretion onto GnRH neurons. However, some aspects of this paradigm remain ill defined. Here we aimed to characterize the following: 1) the effects of NKB signaling on FSH secretion and 2) the role of Dyn in gonadotropin secretion after NK3R activation; 3) additionally, we explored the roles of other tachykinin receptors, NK1R and NK2R, on gonadotropin release. Thus, the effects of the NK3R agonist, senktide, on FSH release were explored across postnatal development in male and female rats; gonadotropin responses to agonists of NK1R substance P and NK2R [neurokinin A (NKA)] were also monitored. Moreover, the effects of senktide on gonadotropin secretion were assessed after antagonizing Dyn actions by nor-binaltorphimine didydrochloride. Before puberty, rats of both sexes showed increased FSH secretion to senktide (and Kp-10). Conversely, adult female rats were irresponsive to senktide in terms of FSH, despite proven LH responses, whereas the adult males did not display FSH or LH responses to senktide, even at high doses. In turn, substance P and NKA stimulated gonadotropin secretion in prepubertal rats, whereas in adults modest gonadotropin responses to NKA were detected. By pretreatment with a Dyn antagonist, adult males became responsive to senktide in terms of LH secretion and displayed elevated basal LH and FSH levels; nor-binaltorphimine didydrochloride treatment uncovered FSH responses to senktide in adult females. Furthermore, the expression of Pdyn and Opkr1 (encoding Dyn and κ-opioid receptor, respectively) in the mediobasal hypothalamus was greater in males than in females at prepubertal ages. Overall, our data contribute to refining our understanding on how the elements of the KNDy node and related factors (ie, other tachykinins) differentially participate in the control of gonadotropins at different stages of rat postnatal maturation.

Metabolic programming of puberty: sexually dimorphic responses to early nutritional challenges.

Body energy stores and metabolic cues influence the onset of puberty. However, the pubertal impact of early nutritional challenges has been only fragmentarily addressed. We evaluated here the consequences, in terms of pubertal timing and hormonal markers, of various nutritional manipulations during pre- or postnatal maturation in rats of both sexes. Males and females were submitted to gestational undernutrition (UNG) or peripubertal (SUB) subnutrition or were raised in large (LL; underfeeding) or small (SL; overfeeding) litters. In addition, groups of UNG, LL, and SL rats were fed on a high-fat diet (HFD) after weaning. Postnatal overfeeding resulted in higher body weights (BWs) during pubertal transition in both sexes, but only SL males displayed overtly advanced external signs of puberty. Postnatal underfeeding persistently decreased BW gain during puberty, yet the magnitude of pubertal delay was greater in LL males. In contrast, regardless of postnatal nutrition, HFD tended to advance the onset of puberty in females but did not alter pubertal timing in males. Likewise, SUB females displayed a marked delay in BW gain and puberty onset, whereas despite similar reduction in BW, SUB males showed normal timing of puberty. These sex divergences were also detected in various hormonal and metabolic indices so that postnatal overnutrition consistently increased LH, FSH, leptin, and insulin levels only in pubertal females, whereas HFD decreased gonadotropin levels in SL females but increased them in SL males. Notably, UNG rats did not show signs of delayed puberty but displayed a striking sex dimorphism in serum insulin/glucose levels, regardless of the diet, so that only UNG males had signs of presumable insulin resistance. Our data disclose important sex differences in the impact of various early nutritional challenges on the timing of puberty, which may help to explain the different trends of altered puberty and related comorbidities between sexes.

Changes in hypothalamic expression of the Lin28/let-7 system and related microRNAs during postnatal maturation and after experimental manipulations of puberty.

Lin28 and Lin28b are related RNA-binding proteins that inhibit the maturation of miRNAs of the let-7 family and participate in the control of cellular stemness and early embryonic development. Considerable interest has arisen recently concerning other physiological roles of the Lin28/let-7 axis, including its potential involvement in the control of puberty, as suggested by genome-wide association studies and functional genomics. We report herein the expression profiles of Lin28 and let-7 members in the rat hypothalamus during postnatal maturation and in selected models of altered puberty. The expression patterns of c-Myc (upstream positive regulator of Lin28), mir-145 (negative regulator of c-Myc), and mir-132 and mir-9 (putative miRNA repressors of Lin28, predicted by bioinformatic algorithms) were also explored. In male and female rats, Lin28, Lin28b, and c-Myc mRNAs displayed very high hypothalamic expression during the neonatal period, markedly decreased during the infantile-to-juvenile transition and reached minimal levels before/around puberty. A similar puberty-related decline was observed for Lin28b in monkey hypothalamus but not in the rat cortex, suggesting species conservation and tissue specificity. Conversely, let-7a, let-7b, mir-132, and mir-145, but not mir-9, showed opposite expression profiles. Perturbation of brain sex differentiation and puberty, by neonatal treatment with estrogen or androgen, altered the expression ratios of Lin28/let-7 at the time of puberty. Changes in the c-Myc/Lin28b/let-7 pathway were also detected in models of delayed puberty linked to early photoperiod manipulation and, to a lesser extent, postnatal underfeeding or chronic subnutrition. Altogether, our data are the first to document dramatic changes in the expression of the Lin28/let-7 axis in the rat hypothalamus during the postnatal maturation and after different manipulations that disturb puberty, thus suggesting the potential involvement of developmental changes in hypothalamic Lin28/let-7 expression in the mechanisms permitting/leading to puberty onset.

Predicción de unión de péptidos de Plasmodium falciparum al HLA clase II. Probabilidad, combinatoria y entropía aplicadas a las proteínas MSP-5 y MSP-6 / Plasmodium falciparum binding peptides prediction to HLA class II. Probability, combinatory and entropy applied to MSP-5 and MSP-6 proteins

Las proteínas de superficie del merozoíto (MSP) son de importancia en la invasión parasitaria al glóbulo rojo. La proteína MSP-5, encontrada en merozoítos libres, tiene un papel en la inmunización de ratones al P. falciparum y P. yoelii, pese a lo cual algunos estudios cuestionan su rol en la invasión. La proteína MSP-6 forma junto con MSP-1 y MSP-7 un complejo en la superficie del merozoíto, liberado del parásito cerca del momento de la invasión al glóbulo rojo. Con el fin de predecir el fenómeno de unión de péptidos de las proteínas de superficie MSP-5 y MSP-6, se aplicó una teoría de unión al HLA clase II, a la totalidad de secuencias de 20 aminoácidos de tales moléculas. Se calcularon los valores de probabilidad, combinatoria y entropía de 168 secuencias nonámeras sobrelapadas de la proteína MSP-5 y 228 de MSP-6. Por último se aplicó la teoría de unión a todos los péptidos nonámeros de tres proteínas construidas computacionalmente, cada una con una longitud de 500 aminoácidos. Para la proteína MSP-5 se predijo un total de 31 secuencias asociadas al macroestado de unión y 137 al de no unión, mientras que se predijo la existencia de 35 secuencias asociadas al macroestado de unión para MSP-6 y 193 al de no unión. Se encontraron respectivamente 100, 111 y 91 secuencias predichas de unión para las tres proteínas teóricas construidas. La predicción teórica de unión de péptidos es útil para facilitar el desarrollo de vacunas, al evidenciar el orden físico-matemático subyacente al fenómeno.(AU)

Neurokinin B and the control of the gonadotropic axis in the rat: developmental changes, sexual dimorphism, and regulation by gonadal steroids.

Neurokinin B (NKB), encoded by Tac2 in rodents, and its receptor, NK3R, have recently emerged as important regulators of reproduction; NKB has been proposed to stimulate kisspeptin output onto GnRH neurons. Accordingly, NKB has been shown to induce gonadotropin release in several species; yet, null or even inhibitory effects of NKB have been also reported. The basis for these discrepant findings, as well as other key aspects of NKB function, remains unknown. We report here that in the rat, LH responses to the NK3R agonist, senktide, display a salient sexual dimorphism, with persistent stimulation in females, regardless of the stage of postnatal development, and lack of LH responses in males from puberty onward. Such dimorphism was independent of the predominant sex steroid after puberty, because testosterone administration to adult females failed to prevent LH responses to senktide, and LH responsiveness was not restored in adult males treated with estradiol or the nonaromatizable androgen, dihydrotestosterone. Yet, removal of sex steroids by gonadectomy switched senktide effects to inhibitory, both in adult male and female rats. Sexual dimorphism was also evident in the numbers of NKB-positive neurons in the arcuate nucleus (ARC), which were higher in adult female rats. This is likely the result of differences in sex steroid milieu during early periods of brain differentiation, because neonatal exposures to high doses of estrogen decreased ARC NKB neurons at later developmental stages. Likewise, neonatal estrogenization resulted in lower serum LH levels that were normalized by senktide administration. Finally, we document that the ability of estrogen to inhibit hypothalamic Tac2 expression seems region specific, because estrogen administration decreased Tac2 levels in the ARC but increased them in the lateral hypothalamus. Altogether, our data provide a deeper insight into relevant aspects of NKB function as major regulator of the gonadotropic axis in the rat, including maturational changes, sexual dimorphism, and differential regulation by sex steroids.

Cellular distribution, regulated expression, and functional role of the anorexigenic peptide, NUCB2/nesfatin-1, in the testis.

Nesfatin-1, product of the precursor NEFA/nucleobindin2 (NUCB2), was initially identified as anorectic hypothalamic neuropeptide, acting in a leptin-independent manner. In addition to its central role in the control of energy homeostasis, evidence has mounted recently that nesfatin-1 is also produced in peripheral metabolic tissues, such as pancreas, adipose, and gut. Moreover, nesfatin-1 has been shown to participate in the control of body functions gated by whole-body energy homeostasis, including puberty onset. Yet, whether, as is the case for other metabolic neuropeptides, NUCB2/nesfatin-1 participates in the direct control of gonadal function remains unexplored. We document here for the first time the expression of NUCB2 mRNA in rat, mouse, and human testes, where NUCB2/nesfatin-1 protein was identified in interstitial mature Leydig cells. Yet in rats, NUCB2/nesfatin-1 became expressed in Sertoli cells upon Leydig cell elimination and was also detected in Leydig cell progenitors. Although NUCB2 mRNA levels did not overtly change in rat testis during pubertal maturation and after short-term fasting, NUCB2/nesfatin-1 content significantly increased along the puberty-to-adult transition and was markedly suppressed after fasting. In addition, testicular NUCB2/nesfatin-1 expression was up-regulated by pituitary LH, because hypophysectomy decreased, whereas human choriogonadotropin (super-agonist of LH receptors) replacement enhanced, NUCB2/nesfatin-1 mRNA and peptide levels. Finally, nesfatin-1 increased human choriogonadotropin-stimulated testosterone secretion by rat testicular explants ex vivo. Our data are the first to disclose the presence and functional role of NUCB2/nesfatin-1 in the testis, where its expression is regulated by developmental, metabolic, and hormonal cues as well as by Leydig cell-derived factors. Our observations expand the reproductive dimension of nesfatin-1, which may operate directly at the testicular level to link energy homeostasis, puberty onset, and gonadal function.

Sex steroids and the control of the Kiss1 system: developmental roles and major regulatory actions.

Kisspeptins, encoded by the Kiss1 gene, and their canonical receptor, GPR54 (also termed Kiss1R), are unanimously recognised as essential regulators of puberty onset and gonadotrophin secretion. These key reproductive functions stem from the capacity of kisspeptins to stimulate gonadotrophin-releasing hormone (GnRH) secretion in the hypothalamus, where discrete populations of Kiss1 neurones have been identified. In rodents, two major groups of hypothalamic Kiss1 neurones exist: one present in the arcuate nucleus (ARC) and the other located in the anteroventral periventricular area (AVPV/RP3V). In recent years, numerous signals have been identified as putative modulators of the hypothalamic Kiss1 system. Among them, the prominent role of sex steroids as being important regulators of Kiss1 neurones has been documented in different species and developmental stages, such as early brain sex differentiation, puberty, adulthood and senescence. These regulatory actions are (mainly) conducted via oestrogen receptor (ER)α, which is expressed in almost all Kiss1 neurones, and likely involve both classical and nonclassical pathways. The regulatory effects of sex steroids are nucleus-specific. Thus, sex steroids inhibit the expression of Kiss1/kisspeptin at the ARC, as a mechanism to conduct their negative-feedback actions on gonadotrophin secretion. By contrast, oestrogens enhance Kiss1 expression at the AVPV/RP3V in rodents, suggesting the involvement of this population in the positive-feedback actions of oestradiol to generate the preovulatory surge of gonadotrophins. In addition, sex steroids have been shown to act post-transcriptionally, modulating GnRH/gonadotrophin responsiveness to kisspeptin. Finally, sex steroids also regulate the expression of co-transmitters of Kiss1 neurones, such as neurokinin B, whose mRNA content in the ARC fluctuates in parallel to that of Kiss1 in response to changes in the circulating levels of sex steroids, therefore suggesting the contribution of this neuropeptide in the feedback control of gonadotrophin secretion. In sum, compelling experimental evidence obtained in different mammalian (and non-mammalian) species, including primates, demonstrates that sex steroids are essential regulators of hypothalamic Kiss1 neurones, which in turn operate as conduits for their effects on GnRH neurones. The physiological relevance of such regulatory phenomena is thoroughly discussed.

Regulation of NKB pathways and their roles in the control of Kiss1 neurons in the arcuate nucleus of the male mouse.

Kisspeptin (Kiss1) and neurokinin B (NKB) (encoded by the Kiss1 and Tac2 genes, respectively) are indispensable for reproduction. In the female of many species, Kiss1 neurons in the arcuate nucleus (ARC) coexpress dynorphin A and NKB. Such cells have been termed Kiss1/NKB/Dynorphin (KNDy) neurons, which are thought to mediate the negative feedback regulation of GnRH/LH secretion by 17ß-estradiol. However, we have less knowledge about the molecular physiology and regulation of Kiss1/Kiss1-expressing neurons in the ARC of the male. Our work focused on the adult male mouse, where we sought evidence for coexpression of these neuropeptides in cells in the ARC, assessed the role of Kiss1 neurons in negative feedback regulation of GnRH/LH secretion by testosterone (T), and investigated the action of NKB on KNDy and GnRH neurons. Results showed that 1) the mRNA encoding Kiss1, NKB, and dynorphin are coexpressed in neurons located in the ARC; 2) Kiss1 and dynorphin A mRNA are regulated by T through estrogen and androgen receptor-dependent pathways; 3) senktide, an agonist for the NKB receptor (neurokinin 3 receptor, encoded by Tacr3), stimulates gonadotropin secretion; 4) KNDy neurons express Tacr3, whereas GnRH neurons do not; and 5) senktide activates KNDy neurons but has no discernable effect on GnRH neurons. These observations corroborate the putative role for KNDy neurons in mediating the negative feedback effects of T on GnRH/LH secretion and provide evidence that NKB released from KNDy neurons is part of an auto-feedback loop that generates the pulsatile secretion of Kiss1 and GnRH in the male.

Acute inflammation reduces kisspeptin immunoreactivity at the arcuate nucleus and decreases responsiveness to kisspeptin independently of its anorectic effects.

Severe inflammatory challenges are frequently coupled to decreased food intake and disruption of reproductive function, the latter via deregulation of different signaling pathways that impinge onto GnRH neurons. Recently, the hypothalamic Kiss1 system, a major gatekeeper of GnRH function, was suggested as potential target for transmitting immune-mediated repression of the gonadotropic axis during acute inflammation, and yet key facets of such a phenomenon remain ill defined. Using lipopolysaccharide S (LPS)-treated male rats as model of inflammation, we document herein the pattern of hypothalamic kisspeptin immunoreactivity (IR) and hormonal responses to kisspeptin during the acute inflammatory phase. LPS injections induced a dramatic but transient drop of serum LH and testosterone levels. Suppression of gonadotropic function was associated with a significant decrease in kisspeptin-IR in the arcuate nucleus (ARC) that was not observed under conditions of metabolic stress induced by 48-h fasting. In addition, absolute responses to kisspeptin-10 (Kp-10), in terms of LH and testosterone secretion, were significantly attenuated in LPS-treated males that also displayed a decrease in food intake and body weight. Yet pair-fed males did not show similar alterations in LH and testosterone secretory responses to Kp-10, whose magnitude was preserved, if not augmented, during food restriction. In summary, our data document the impact of acute inflammation on kisspeptin content at the ARC as key center for the neuroendocrine control of reproduction. Our results also suggest that suppressed gonadotropic function following inflammatory challenges might involve a reduction in absolute responsiveness to kisspeptin that is independent of the anorectic effects of inflammation.