Búsqueda de marcadores de embarazo ectópico en sangre periférica

Motivación: El embarazo ectópico es una anomalía en la implantación embrionaria y complicación ginecológica grave que afecta a más de un 1% de los embarazos naturales y hasta un 8% de las embarazadas en reproducción asistida, según las fuentes. Los métodos diagnósticos de embarazo ectópico a día de hoy se basan exclusivamente en métodos ecográficos y en medidas seriadas de B-hCG métodos que, en muchas ocasiones, se prolongan por una semana o más, para llegar a un diagnóstico fiable.En el Reino Unido el diagnóstico del embarazo ectópico genera un gasto anual de 12 millones de euros², datos extrapolables al resto de Europa. La existencia de un buen marcador de embarazo ectópico sustituiría la realización del test diario de β-hCG, siendo el mercado potencial de unos 13 millones de análisis por año.Métodos: Se recogieron seis muestras de sangre procedentes de mujeres con embarazo ectópico y otras seis de mujeres con embarazo intrauterino con edades gestacionales similares (entre 7 y 10 semanas) se almacenaron a -20ºC en tubos PAXGene hasta su posterior análisis. La extracción del RNA se realizó utilizando el método del Trizol (Sigma-Aldrich). Todas las muestras fueron hibridadas utilizando el “Whole Human Genome Oligo Microarray” (Agilent Technologies) que contiene mas de 44.000 sondas para el genoma humano. El microarray hibridado se escaneo mediante el escaner Axon 4100 (Molecular Devices) y los datos se extrajeron con el software Genepix 6.0 (Molecular Devices). El análisis de datos se llevo acabo con la plataforma GEPAS. Resultados: Se utilizaron seis muestras de embarazo ectópico y seis muestras de embarazo intrauterino para el análisis de microarray. Se generó una lista de 17 potenciales marcadores de embarazo ectópico con un fold change >4 o <-4 y con un p valor <0.05. De estos 17 genes se han validados por PCR cuantitativa los siguientes: ORM1, ORM2, LAMP3 y UTS2 utilizando nuevas muestras de embarazo ectópico (n=8) y de embarazo intrauterino (n=7). Después de descartar los valores atípicos se confirmaron algunos resultados del microarray.Conclusiones: Los resultados del microarray mostraron un total de 17 genes desregulados entre embarazos ectópicos e intrauterinos, concretamente 10 se sobre expresaban y 7 de ellos disminuían su expresión. Los genes ORM1, ORM2, LAMP3 y UTS2 se comprobaron por qPCR y mostraron diferencias significativas. . Al final de este estudio se comprobaran el resto de genes y se podrá diseñar un test de embarazo ectópico

Incidence of uterine post abortion infection at Hospital de Clínicas de Porto Alegre. Is prophylactic antibiotic necessary?

Objetivo: Identificar a incidência de infecção pélvica após aborto espontâneo submetido a esvaziamento uterino num hospital terciário do sul do Brasil e comparar com a literatura internacional. Métodos: Os prontuários eletrônicos do Hospital de Clínicas de Porto Alegre de todas as pacientes que foram submetidas ao esvaziamento uterino por abortamento entre agosto de 2008 e Janeiro de 2012 foram revisados. Foram incluídas no estudo todas as pacientes submetidas à curetagem uterina por abortamento e que tiveram consultas ambulatoriais de revisão após o procedimento. Os dados demográficos e laboratoriais da população estudada, number needed for treatment (NNT) e o number needed to harm (NNH) foram calculados. Resultados: Dos 857 prontuários eletrônicos revistos, 377 pacientes foram submetidas ao esvaziamento uterino por abortamento; 55 casos foram perdidos no seguimento, restando 322 casos que foram classificados como aborto não infectado na admissão. A maioria da população era da raça branca (79%); a prevalência de HIV e VDRL positivos foi de 0,3 e 2%, respectivamente. No seguimento desses 322 casos, num período mínimo de 7 dias, verificou-se que a incidência de infecção pós-procedimento foi de 1,8% (IC95%0,8 a 4). O NNT e o NNH calculado para 42 meses foi de 63 e 39, respectivamente. Conclusão: A incidência de infecção pós-aborto entre agosto de 2008 a janeiro de 2012 foi de 1,8% (0,8 a 4).Objective: To identify the incidence of infection post-uterine evacuation for miscarriage at a tertiary teaching hospital in southern Brazil. Methods: Electronic records of all patients admitted for uterine evacuation for miscarriage between August 2008 and January 2012 were revised. All patients submitted to uterine curettage for miscarriage and had outpatient follow-up were included. Demographic, laboratorial data of the sampled population, the number needed to treat (NNT) and number needed to harm (NNH) were calculated. Results: From 857 reviewed electronic records, 377 underwent uterine evacuation for miscarriage. 55 cases were lost to follow-up, remaining 322 cases classified as non-infected miscarriage at admission. The majority of the population was white (79%); prevalence of positive HIV and VDRL was 0.3 and 2%, respectively. From 322 cases, within a week of follow-up, the incidence of post-miscarriage infection was 1.8% (95%CI 0.8 to 4). In a period of 42 months, the NNT and NNH were 63 and 39, respectively. Conclusion: The incidence of infection after miscarriage between August 2008 and January 2012 was 1.8% (0.8 to 4)

Ectopic Pregnancy as a Model to Identify Endometrial Genes and Signaling Pathways Important in Decidualization and Regulated by Local Trophoblast

The endometrium in early pregnancy undergoes decidualization and functional changes induced by local trophoblast, which are not fully understood. We hypothesized that endometrium from tubal ectopic pregnancy (EP) could be interrogated to identify novel genes and pathways involved in these processes. Gestation-matched endometrium was collected from women with EP (n = 11) and intrauterine pregnancies (IUP) (n = 13). RNA was extracted from the tissue. In addition, tissues were prepared for histological analysis for degree of decidualization. We compared a) the samples from EP that were decidualized (n = 6) with non-decidualized samples (n = 5), and b) the decidualized EP (n = 6) with decidualization-matched IUP (n = 6) samples using an Affymetrix gene array platform, with Ingenuity Pathway Analysis, combined with quantitative RT-PCR. Expression of PRL and IGFBP1 was used to confirm the degree of decidualization in each group. There were no differences in PRL or IGFBP1 expression in the decidualization-matched samples but a marked reduction (P<0.001) in the non-decidualized samples. Decidualization was associated with increased expression of 428 genes including SCARA5 (181-fold), DKK1 (71-fold) and PROK1 (32-fold), and decreased expression of 230 genes including MMP-7 (35-fold) and SFRP4 (21-fold). The top canonical pathways associated with these differentially expressed genes were Natural Killer Cell and Wnt/b-Catenin signaling. Local trophoblast was associated with much less alteration of endometrial gene expression with an increase in 56 genes, including CSH1 (8-fold), and a reduction in 29 genes including CRISP3 (8-fold). The top associated canonical pathway was Antigen Presentation. The study of endometrium from tubal EP may promote novel insights into genes involved in decidualization and those influenced by factors from neighboring trophoblast. This has afforded unique information not highlighted by previous studies and adds to our understanding of the endometrium in early pregnancy

Postoperative intravenously administered iron sucrose versus postoperative orally administered iron to treat post-bariatric abdominoplasty anaemia (ISAPA): the study protocol for a randomised controlled trial

BACKGROUND: Anaemia and iron deficiency are common complications following post-bariatric abdominoplasty. Given the low oral absorbability of iron resulting from bariatric surgery, it has been hypothesised that postoperative intravenously administered iron supplementation could be used to treat anaemia and to prevent the development of iron deficiency in these patients. METHODS/DESIGN: In this multicentre open-label randomised clinical trial, 56 adult women undergoing post-bariatric anchor-line abdominoplasty will be allocated at a ratio of 1:1 for postoperative supplementation with two intravenously administered applications of 200 mg of iron saccharate or postoperative supplementation with 100 mg of iron polymaltose complex administered orally, twice a day for 8 weeks. The primary outcome is the difference in mean haemoglobin levels between the two groups at eight postoperative weeks. Secondary outcomes evaluated at one, four and eight postoperative weeks include iron profile, reticulocyte count, overall quality of life measured using the Short-Form 36 Health Survey (SF-36) questionnaire, fatigue measured using the Functional Assessment of Chronic Illness Therapy – Fatigue (FACIT-F), adverse effects and postoperative complications. DISCUSSION: This randomised clinical trial aims to evaluate the haematopoietic effectiveness of intravenously administered iron supplementation in patients undergoing post-bariatric abdominoplasty. A more effective recovery of haemoglobin levels could help improve the patients’ quality of life and could provide an improved haematological status in preparation for the subsequent and frequent plastic surgeries these patients undergo. TRIAL REGISTRATION: Clinicaltrials.gov Identifier: NCT01857011 (8 May 2013), Universal Trial Number U111-1169-6223, Brazilian Clinical Trials Registry (REBEC): RBR-2JGRKQ