Oncogénesis del pólipo endometrial. Manejo clínico a través de marcadores biomoleculares / Oncogenesis of the endometrial polyp. Clinical management through biomolecular markers

La prevalencia de los pólipos endometriales se estima entre el 7,8 y el 35% de las mujeres, siendo mayor en el estado posmenopáusico. Los pólipos endometriales se asocian con hiperplasia endometrial y carcinogénesis, con una prevalencia informada de lesiones malignas y premalignas que puede llegar al 13%. La detección de pólipos endometriales en edad peri o posmenopáusica, en pacientes sintomáticas o asintomáticas, requiere un examen histeroscópico meticuloso. No está claro si la polipectomía debe realizarse de forma rutinaria en pacientes asintomáticas. El manejo expectante de pólipos pequeños y asintomáticos es razonable en muchos casos. Se necesitan estudios adicionales para dilucidar si los pólipos endometriales son precursores de cáncer, o simplemente marcadores de una enfermedad endometrial. Los biomarcadores capaces de detectar cambios a nivel molecular en los pólipos y el tejido endometrial nos ayudan a un mejor conocimiento y clasificación de los procesos malignos. Este conocimiento permite pasar de una medicina intervencionista a una medicina más conservadora, basada en la confianza de un conocimiento más preciso de los procesos biomoleculares. GynEC®-DX se basa en determinar la expresión de genes que se combinan en un algoritmo matemático diagnóstico para llegar a un diagnóstico negativo o positivo de cáncer de endometrio. La prueba molecular tiene un valor predictivo negativo del 99,6%, con una alta especificidad y sensibilidad. Esta prueba podría usarse para el diagnóstico diferencial del cáncer de endometrio en mujeres con pólipos endometriales sin requerir la exéresis de pólipos, limitando el riesgo iatrogénico y evitando intervenciones innecesarias

The prevalence of endometrial polyps is estimated between 7.8 and 35% of women, being more prevalent in postmenopausal women. Endometrial polyps are associated with endometrial hyperplasia and carcinogenesis, with an informed prevalence of malignant and premalignant lesions that may reach 13%. The detection of endometrial polyps in peri- or postmenopausal (status) age, in symptomatic or asymptomatic patients, requires a meticulous hysteroscopic examination. It is unclear if routine polypectomy should be performed in asymptomatic patients. The expectant management of small and asymptomatic polyps is reasonable in many cases. Additional studies are needed to elucidate whether endometrial polyps are precursors of cancer, or simply markers of an endometrial disease. Biomarkers capable of detecting changes at the molecular level in polyps and endometrial tissue help us to better understand and classify malignant processes. This knowledge allows to move from an interventional medicine to a more conservative medicine, based on the confidence of a more precise knowledge of the biomolecular processes. GynEC®-DX is based on determining the expression of genes that are combined in a diagnostic mathematical algorithm to arrive at a negative or positive diagnosis of endometrial cancer. The molecular test has a negative predictive value of 99.6%, with high specificity and sensitivity. This test could be used for the differential diagnosis of endometrial cancer in women with EPs and prevent the resection of polyps, limiting the iatrogenic risk and avoiding unnecessary interventions

Ambulatory MiniArc Precise Sling under Local Anesthesia for Stress Urinary Incontinence: Feasibility and Outcome.

AIMS: The aim of the study is to assess the feasibility of ambulatory stress urinary incontinence (SUI) surgery using the MiniArc Precise single-incision urethral sling without increasing the number of complications. SETTINGS AND DESIGN: This was a retrospective observational study of prospectively collected data carried out in a Tertiary Referral Hospital in Barcelona, Spain. MATERIALS AND METHODS: Forty patients diagnosed with SUI or stress predominant mixed urinary incontinence (MUI) treated surgically between November 2011 and November 2013. The MiniArc Precise® sling was inserted under local anesthesia in the ambulatory setting. STATISTICAL ANALYSIS USED: Descriptive statistics included frequencies and percentages for categorical variables and mean and range for quantitative variables. The statistical package used was SPSS version 17.0. RESULTS: Urodynamic studies showed SUI in 78% of cases and stress predominant MUI in 17%. Clinical findings included SUI in 56% of cases and MUI in 44%, with positive stress tests in all participants. The mean intraoperative pain (1-10 Visual Analog Scale) was 2. All patients were satisfied with the use of local anesthesia in the outpatient setting. Perioperative complications did not occur. One case of urinary retention and two cases of urinary tract infection (UTI) developed within this 1st month after operation and were successfully managed conservatively. Midterm complications included eight cases of UTI and four de novo urge incontinence. CONCLUSIONS: Placement of the MiniArc Precise sling under local anesthesia is a feasible and safe technique, which when carried out by an experienced surgeon allows to be done as an outpatient basis without increasing the rate of postprocedural complications.

Diagnóstico molecular del cáncer de endometrio / Molecular diagnosis of endometrial cancer

Mediante caracterización molecular se han identificado una combinación de biomarcadores fuertemente relacionados con la presencia de cáncer de endometrio. Concretamente, se ha desarrollado un test de diagnóstico molecular asociado a un algoritmo matemático que, con el análisis de una simple muestra de aspirado endometrial, permite mejorar el diagnóstico precoz del cáncer de endometrio. Este test, llamado GynEC(R)-DX, se basa en la interpretación de cambios moleculares que preceden las alteraciones morfológicas asociadas al cáncer de endometrio. La aplicabilidad del test GynEC(R)-DX, rápido y fiable, mejora por sí mismo la precisión diagnóstica de las pruebas convencionales de biopsia por aspirado o dirigida por histeroscopia, minimizando la posibilidad de resultados no concluyentes, y aporta una información de alto valor en determinados perfiles clínicos, que refuerza la confianza en la toma de decisión terapéutica y en el proceso de despistaje del cáncer de endometrio. Consecuentemente, se reduce el tiempo y el coste medio del diagnóstico de cáncer, especialmente en 5 perfiles específicos de pacientes característicos: hemorragia uterina anormal de repetición, ecografía con línea endometrial irregular sin dictamen claro histológico, diagnóstico de pólipo endometrial, hiperplasia endometrial sin atipias y Síndrome de Lynch. En el presente trabajo se describe el momento de aplicación del diagnóstico molecular, adaptado al algoritmo diagnóstico de la SEGO, para cada uno de los citados perfiles de alto riesgo, así como las ventajas clínicas derivadas (AU)

Through molecular characterization, a combination of biomarkers strongly related to the presence of endometrial cancer has been identified. Specifically, a molecular diagnostic test associated with a mathematical algorithm has been developed which, with the analysis of a simple sample of endometrial aspirate, allows improving the early diagnosis of endometrial cancer. This test, called GynEC(R)-DX, is based on the interpretation of molecular changes that precede the morphological alterations associated with endometrial cancer. The applicability of this test, called GynEC(R)-DX, fast and reliable, improves by itself the diagnostic accuracy of conventional aspiration or hysteroscopy-guided biopsy tests, minimizing the possibility of inconclusive results, and provides high value information in certain clinical profiles, which reinforces confidence in therapeutic decision- making and in the endometrial cancer screening process. The combination of GynEC(R)-DX with histological analysis on endometrial aspirate increases efficacy, sensitivity and Negative Predictive Value. Consequently, time and average cost of cancer diagnosis is reduced, especially in 5 characteristic profiles of abnormal patients: recurrent abnormal uterine bleeding, ultrasound with irregular endometrial line without clear histological opinion, diagnosis of endometrial polyp, endometrial hyperplasia without atypia and Lynch Syndrome. The present work describes the application of endometrial cancer molecular diagnosis, adapted to the SEGO diagnostic algorithm, for the high risk profiles mentioned above, as well as the derived clinical advantages (AU)

Preliminary results: ethanol sclerotherapy after ultrasound-guided fine needle aspiration without anesthesia in the management of simple ovarian cysts.

STUDY OBJECTIVE: To investigate the feasibility of ultrasound-guided, fine-needle aspiration and ethanol sclerotherapy of simple ovarian cysts in an office setting without anesthesia. We also describe the rate of cyst recurrence in patients treated with this technique and explore the associated risk factors. DESIGN: Prospective follow-up of patients after ethanol sclerotherapy of simple adnexal cysts in a single center trial (Canadian Task Force classification II-1). SETTING: The study was conducted at Bellvitge Teaching Hospital in Barcelona, Spain. PATIENTS: Ethanol sclerotherapy was performed on 60 simple adnexal cysts between 2009 and 2012. INTERVENTIONS: Ultrasound-guided fine-needle aspiration and ethanol sclerotherapy. MEASUREMENTS AND MAIN RESULTS: Patient demographics and cyst characteristics were collected for all patients. Potential risk factors for recurrence were analyzed by univariate and multivariate analyses. All the procedures, except 1, were performed without anesthesia. The only major complication was a case of self-limiting hem peritoneum that was managed expectantly. Moderate abdominal pain occurred in 26.7% of patients during the procedure. Fifty-five patients completed at least 6 months of follow-up and were included in the statistical recurrence analyses. Cyst recurrence was recorded in 9.1% of the patients and was managed in the usual manner in all the cases. Univariate analyses indicated that a larger cyst diameter and a higher estimated cyst volume were significantly associated with recurrence. In the multivariate analysis, recurrence was only significantly associated with estimated cyst volume. CONCLUSION: Ethanol sclerotherapy of simple ovarian cysts in an office setting without anesthesia is a feasible technique associated with a low rate of complications and recurrence, although larger randomized studies are necessary to assess the predictive factors for cyst recurrence.

Guía de cribado del cáncer de cuello de útero en España, 2014 / No disponible

No disponible

Guía de cribado del cáncer de cuello de útero en España, 2014 / No disponible

No disponible

A novel IRAK1-IKKε signaling axis limits the activation of TAK1-IKKß downstream of TLR3.

IRAK1 is involved in the regulation of type I IFN production downstream of TLR3. Previous work indicated that IRAK1 negatively regulates TRIF-mediated activation of IRF3 and IRF7. We report that IRAK1 limits the activation of the TLR3-NF-κB pathway. Following TLR3 stimulation, IRAK1-deficient macrophages produced increased levels of IL-6 and IFN-ß compared with wild type macrophages. Pharmacological inhibition of TAK1 reduced this increase in IFN-ß, together with the heightened activation of IRF3 and p65 found in TLR3-ligand stimulated IRAK1-deficient macrophages. Recently, IKKε and TANK-binding kinase 1 (TBK1) were reported to limit activation of the NF-κB pathway downstream of IL-1R, TNFR1, and TLRs. We show that TBK1 has a positive role in the TLR3-NF-κB pathway, because we detected reduced levels of IL-6 and reduced activation of p65 in TBK1-deficient macrophages. In contrast, we show that IKKε limits the activation of the TLR3-NF-κB pathway. Furthermore, we show that IRAK1 is required for the activation of IKKε downstream of TLR3. We report impaired activation of ERK1/2 in IRAK1- and IKKε-deficient macrophages, a novel finding for both kinases. Importantly, this work provides novel mechanistic insight into the regulation of the TLR3-signaling pathway, providing strong evidence that an IRAK1-IKKε-signaling axis acts to limit the production of both type I IFNs and proinflammatory cytokines by regulating TAK1 activity.

Dimerization of Smac is crucial for its mitochondrial retention by XIAP subsequent to mitochondrial outer membrane permeabilization.

Following the apoptotic permeabilization of the outer mitochondrial membrane, the inter-membrane space protein second mitochondria-derived activator of caspases (Smac) is released into the cytosol. Smac efficiently promotes apoptosis by antagonizing x-linked inhibitor of apoptosis protein (XIAP), an inhibitor of caspases-9, -3, and -7, via a short NH(2)-terminal inhibitor of apoptosis protein (IAP) binding motif (AVPI). Native Smac dimerizes to form a highly stable and inflexible elongated arch, however, a functional role for this outstretched structure so far remained unknown. Using time-lapse single-cell imaging of DLD-1 and HCT-116 colon cancer cells, we here demonstrate that upon mitochondrial outer membrane permeabilization physiological expression levels of XIAP are sufficient to selectively prolong the release of dimeric but not monomeric Smac. Elevating the expression of XIAP further extended the release duration of dimeric Smac and resulted in the mitochondrial retention of a significant proportion of the Smac pool. In contrast, monomeric Smac was always fully released and the release kinetics were not affected by altered XIAP expression. Our findings therefore indicate that the dimerization of Smac is critical for the XIAP-mediated retention of Smac at or inside the mitochondria. This article is part of a Special Issue entitled: 11th European Symposium on Calcium.

Control of motoneuron survival by angiogenin.

Mutations in the hypoxia-inducible factor angiogenin (ANG) have been identified in Amyotrophic Lateral Sclerosis (ALS) patients, but the potential role of ANG in ALS pathogenesis was undetermined. Here we show that angiogenin promotes motoneuron survival both in vitro and in vivo. Angiogenin protected cultured motoneurons against excitotoxic injury in a PI-3-kinase/Akt kinase-dependent manner, whereas knock-down of angiogenin potentiated excitotoxic motoneuron death. Expression of wild-type ANG protected against endoplasmic reticulum (ER) stress-induced and trophic-factor-withdrawal-induced cell death in vitro, whereas the ALS-associated ANG mutant K40I exerted no protective activity and failed to activate Akt-1. In SOD1(G93A) mice angiogenin delivery increased lifespan and motoneuron survival, restored the disease-associated decrease in Akt-1 survival signaling, and reversed a pathophysiological increase in ICAM-1 expression. Our data demonstrate that angiogenin is a key factor in the control of motoneuron survival.

Molecular characterization of a glycerophosphoinositol transporter in mammalian cells.

The glycerophosphoinositols are ubiquitous phosphoinositide metabolites involved in the control of several cell functions. They exert their actions both intracellularly and by rapidly equilibrating across the plasma membrane when added to cells, implying the existence of a transporter for their membrane permeation. Such a transporter, GIT1, has been cloned in yeast. By PSI-BLAST analysis, we have identified the Glut2 transporter as a human-genome candidate ortholog of GIT1. This was supported directly through the use of inhibitors, siRNAs and competition studies of specific uptake of GroPIns in HeLa cells over-expressing human Glut2. These data identify Glut2 as a GroPIns transporter in mammals, and define a physiologically relevant cell-permeation mechanism.

A novel pathway of cell growth regulation mediated by a PLA2alpha-derived phosphoinositide metabolite.

The phosphoinositides have well-defined roles in the control of cellular functions, including cytoskeleton dynamics, membrane trafficking, and cell signaling. However, the interplay among the phosphoinositides and their diffusible derivatives that originate through phospholipase A2 action (the lysophosphoinositides and glycerophosphoinositols) remains to be fully elucidated. Here we demonstrate that in PCCl3 rat thyroid cells, the intracellular levels of glycerophosphoinositol are finely modulated by ATP and norepinephrine through the P2Y metabotropic and alpha-adrenergic receptors, respectively. The enzyme involved here is phospholipase A2 IValpha (PLA2 IValpha), which in these cells specifically hydrolyzes phosphatidylinositol, forming lysophosphatidylinositol, glycerophosphoinositol, and arachidonic acid. This receptor-mediated activation of PLA2 IValpha leads to stimulation of PCCl3 cell growth. The involvement of a PLA2 IValpha-mediated pathway is demonstrated by inhibition of the increase in intracellular glycerophosphoinositol levels and cell proliferation by specific inhibitors, RNA interference, and overexpression of the dominant-negative PLA2 IValpha(1-522). Modulation of PCCl3 cell growth is not seen with inhibitors of arachidonic acid metabolism. In conclusion, these data characterize glycerophosphoinositol as a mediator of the purinergic and adrenergic regulation of PCCl3 cell proliferation, defining a novel regulatory cascade specifically involving this soluble phosphoinositide derivative and widening the involvement of the phosphoinositides in the regulation of cell function.

Down's syndrome astrocytes have greater antioxidant capacity than euploid astrocytes.

Down's syndrome (trisomy 21) brain tissue is considered to be susceptible to oxidative injury, mainly because its increased Cu/Zn-superoxide dismutase (SOD1) activity is not followed by an adaptive rise in hydrogen peroxide metabolizing enzymes. In vitro, trisomic neurons suffer oxidative stress and degenerate. We studied the response of trisomy 21 neuron and astrocyte cultures to hydrogen peroxide injury and found that they were, respectively, more and less vulnerable than their euploid counterparts. Differences were detected 24 h after exposures in the region of 50 microm and 500 microm hydrogen peroxide for neuron and astrocyte cultures, respectively. Cytotoxicity results were paralleled by a decrease in cellular glutathione. In addition, trisomic astrocytes showed a lower basal content of superoxide ion and a higher clearance of hydrogen peroxide from the culture medium. In the presence of hydrogen peroxide, trisomic astrocytes maintained their concentration of intracellular superoxide and hydroperoxides at a lower level than euploid astrocytes. Consistent with these results, trisomic astrocytes in neuron coculture were more neuroprotective than euploid astrocytes against hydrogen peroxide injury. We suggest that SOD1 overexpression has beneficial effects on astrocytes, as it does in other systems with similarly high disposal of hydroperoxides. In addition to a higher enzymatic activity of SOD1, cultures of trisomic astrocytes showed slightly higher glutathione reductase activity than euploid cultures. Thus, trisomy 21 astrocytes showed a greater antioxidant capacity against hydrogen peroxide than euploid astrocytes, and they partially counteracted the oxidative vulnerability of trisomic neurons in culture.

Neurotoxicity of organomercurial compounds.

Mercury is a ubiquitous contaminant, and a range of chemical species is generated by human activity and natural environmental change. Elemental mercury and its inorganic and organic compounds have different toxic properties, but all them are considered hazardous in human exposure. In an equimolecular exposure basis, organomercurials with a short aliphatic chain are the most harmful compounds and they may cause irreversible damage to the nervous system. Methylmercury (CH(3)Hg(+)) is the most studied following the neurotoxic outbreaks identified as Minamata disease and the Iraq poisoning. The first description of the CNS pathology dates from 1954. Since then, the clinical neurology, the neuropathology and the mechanisms of neurotoxicity of organomercurials have been widely studied. The high thiol reactivity of CH(3)Hg(+), as well as all mercury compounds, has been suggested to be the basis of their harmful biological effects. However, there is clear selectivity of CH(3)Hg(+) for specific cell types and brain structures, which is not yet fully understood. The main mechanisms involved are inhibition of protein synthesis, microtubule disruption, increase of intracellular Ca(2+) with disturbance of neurotransmitter function, oxidative stress and triggering of excitotoxicity mechanisms. The effects are more damaging during CNS development, leading to alterations of the structure and functionality of the nervous system. The major source of CH(3)Hg(+) exposure is the consumption of fish and, therefore, its intake is practically unavoidable. The present concern is on the study of the effects of low level exposure to CH(3)Hg(+) on human neurodevelopment, with a view to establishing a safe daily intake. Recommendations are 0.4 micro g/kg body weight/day by the WHO and US FDA and, recently, 0.1 micro g/kg body weight/day by the US EPA. Unfortunately, these levels are easily attained with few meals of fish per week, depending on the source of the fish and its position in the food chain.

Evaluation of fluorescent dyes for measuring intracellular glutathione content in primary cultures of human neurons and neuroblastoma SH-SY5Y.

BACKGROUND: Reduced glutathione (GSH) protects cells against oxidative injury and maintains a range of vital functions. To study GSH content in human neuronal cell cultures, thiol-sensitive fluorescent techniques requiring a small number of cells may be of great value, but their GSH specificity has not been established in these cells. METHODS: We tested the efficiency of four currently available GSH fluorescent stains in human neurons and SH-SY5Y neuroblastoma cells, both cultured in microwells, by using a fluorescence plate reader. Cultures were treated with the inhibitor of the GSH synthesis, buthionine sulfoximine (BSO), and progressive GSH depletion was assayed with monochlorobimane (mBCl), monobromobimane (mBBr), 5-chloromethylfluorescein diacetate (CMFDA), and 7-amino-4-chloromethylcoumarin (CMAC). GSH was also determined by a biochemical method in cell homogenates to obtain quantitative reference values. RESULTS: Neurons and SH-SY5Y neuroblastoma had basal GSH contents of 27.1 +/- 3.2 and 14.5 +/- 1.7 nmol/mg protein (n = 5), respectively. An approximate 90% depletion of GSH was obtained after 3 days of exposure to 1,000 microM of BSO in neurons and after 1 day in SH-SY5Y cells. Cell death through an apoptotic pathway appeared 1-2 days after total GSH depletion. The assayed stains had different degrees of background fluorescence and sensitivity to GSH content, with similar results in both neuronal cell types. The probes mBCl and CMAC showed the lowest background, and the GSH-depletion curves were most similar to that of the reference method. CONCLUSIONS: Both mBCl and CMAC are useful fluorescent stains to determine semiquantitative GSH concentration in human neuronal cell cultures.

Cáncer de cérvix. ¿Enfermedad oncológica o enfermedad infecciosa? / No disponible

No disponible