Detection of bile acids in bronchoalveolar lavage fluid defines the inflammatory and microbial landscape of the lower airways in infants with cystic fibrosis.

BACKGROUND: Cystic Fibrosis (CF) is a genetic condition characterized by neutrophilic inflammation and recurrent infection of the airways. How these processes are initiated and perpetuated in CF remains largely unknown. We have demonstrated a link between the intestinal microbiota-related metabolites bile acids (BA) and inflammation in the bronchoalveolar lavage fluid (BALF) from children with stable CF lung disease. To establish if BA indicate early pathological processes in CF lung disease, we combined targeted mass spectrometry and amplicon sequencing-based microbial characterization of 121 BALF specimens collected from 12-month old infants with CF enrolled in the COMBAT-CF study, a multicentre randomized placebo-controlled clinical trial comparing azithromycin versus placebo. We evaluated whether detection of BA in BALF is associated with the establishment of the inflammatory and microbial landscape of early CF lung disease, and whether azithromycin, a motilin agonist that has been demonstrated to reduce aspiration of gastric contents, alters the odds of detecting BA in BALF. We also explored how different prophylactic antibiotics regimens impact the early life BALF microbiota. RESULTS: Detection of BA in BALF was strongly associated with biomarkers of airway inflammation, more exacerbation episodes during the first year of life, increased use of oral antibiotics with prolonged treatment periods, a higher degree of structural lung damage, and distinct microbial profiles. Treatment with azithromycin, a motilin agonist, which has been reported to reduce aspiration of gastric contents, did not reduce the odds of detecting BA in BALF. Culture and molecular methods showed that azithromycin does not alter bacterial load or diversity in BALF. Conversely, penicillin-type prophylaxis reduced the odds of detecting BAs in BALF, which was associated with elevated levels of circulating biomarkers of cholestasis. We also observed that environmental factors such as penicillin-type prophylaxis or BAs detection were linked to distinct early microbial communities of the CF airways, which were associated with different inflammatory landscapes but not with structural lung damage. CONCLUSIONS: Detection of BA in BALF portend early pathological events in CF lung disease. Benefits early in life associated with azithromycin are not linked to its antimicrobial properties. Video Abstract.

Installing oncofertility programs for common cancers in limited resource settings (Repro-Can-OPEN Study): An extrapolation during the global crisis of Coronavirus (COVID-19) pandemic.

PURPOSE: The state of limited resource settings that Coronavirus (COVID-19) pandemic has created globally should be taken seriously into account especially in healthcare sector. In oncofertility, patients should receive their fertility preservation treatments urgently even in limited resource settings before initiation of anticancer therapy. Therefore, it is very crucial to learn more about oncofertility practice in limited resource settings such as in developing countries that suffer often from shortage of healthcare services provided to young patients with cancer. METHODS: As an extrapolation during the global crisis of COVID-19 pandemic, we surveyed oncofertility centers from 14 developing countries (Egypt, Tunisia, Brazil, Peru, Panama, Mexico, Colombia, Guatemala, Argentina, Chile, Nigeria, South Africa, Saudi Arabia, and India). Survey questionnaire included questions on the availability and degree of utilization of fertility preservation options in case of childhood cancer, breast cancer, and blood cancer. RESULTS: All surveyed centers responded to all questions. Responses and their calculated oncofertility scores showed different domestic standards for oncofertility practice in case of childhood cancer, breast cancer, and blood cancer in the developing countries under limited resource settings. CONCLUSIONS: Medical practice in limited resource settings has become a critical topic especially after the global crisis of COVID-19 pandemic. Understanding the resources necessary to provide oncofertility treatments is important until the current COVID-19 pandemic resolves. Lessons learned will be valuable to future potential worldwide disruptions due to infectious diseases or other global crises.

DNA methylation and mRNA expression of imprinted genes in blastocysts derived from an improved in vitro maturation method for oocytes from small antral follicles in polycystic ovary syndrome patients.

STUDY QUESTION: Does imprinted DNA methylation or imprinted gene expression differ between human blastocysts from conventional ovarian stimulation (COS) and an optimized two-step IVM method (CAPA-IVM) in age-matched polycystic ovary syndrome (PCOS) patients? SUMMARY ANSWER: No significant differences in imprinted DNA methylation and gene expression were detected between COS and CAPA-IVM blastocysts. WHAT IS KNOWN ALREADY: Animal models have revealed alterations in DNA methylation maintenance at imprinted germline differentially methylated regions (gDMRs) after use of ARTs. This effect increases as more ART interventions are applied to oocytes or embryos. IVM is a minimal-stimulation ART with reduced hormone-related side effects and risks for patients. CAPA-IVM is an improved IVM system that includes a pre-maturation step (CAPA), followed by an IVM step, both in the presence of physiological compounds that promote oocyte developmental capacity. STUDY DESIGN, SIZE, DURATION: For DNA methylation analysis 20 CAPA-IVM blastocysts were compared to 12 COS blastocysts. For RNA-Seq analysis a separate set of 15 CAPA-IVM blastocysts were compared to 5 COS blastocysts. PARTICIPANTS/MATERIALS, SETTING, METHODS: COS embryos originated from 12 patients with PCOS (according to Rotterdam criteria) who underwent conventional ovarian stimulation. For CAPA-IVM 23 women were treated for 3-5 days with highly purified hMG (HP-hMG) and no hCG trigger was given before oocyte retrieval. Oocytes were first cultured in pre-maturation medium (CAPA for 24 h containing C-type natriuretic peptide), followed by an IVM step (30 h) in medium containing FSH and Amphiregulin. After ICSI, Day 5 or 6 embryos in both groups were vitrified and used for post-bisulphite adaptor tagging (PBAT) DNA methylation analysis or RNA-seq gene expression analysis of individual embryos. Data from specific genes and gDMRs were extracted from the PABT and RNA-seq datasets. MAIN RESULTS AND THE ROLE OF CHANCE: CAPA-IVM blastocysts showed similar rates of methylation and gene expression at gDMRs compared to COS embryos. In addition, expression of major epigenetic regulators was similar between the groups. LIMITATIONS, REASONS FOR CAUTION: The embryos from the COS group were generated in a range of culture media. The CAPA-IVM embryos were all generated using the same sperm donor. The DNA methylation level of gDMRs in purely in vivo-derived human blastocysts is not known. WIDER IMPLICATIONS OF THE FINDINGS: A follow-up of children born after CAPA-IVM is important as it is for other new ARTs, which are generally introduced into clinical practice without prior epigenetic safety studies on human blastocysts. CAPA-IVM opens new perspectives for patient-friendly ART in PCOS. STUDY FUNDING/COMPETING INTEREST(S): IVM research at the Vrije Universiteit Brussel has been supported by grants from the Institute for the Promotion of Innovation by Science and Technology in Flanders (Agentschap voor Innovatie door Wetenschap en Technologie-IWT, project 110680), the Fund for Research Flanders (Fonds voor Wetenschappelijk Onderzoek-Vlaanderen-FWO-AL 679 project, project G.0343.13), the Belgian Foundation Against Cancer (HOPE project, Dossier C69Ref Nr 2016-119) and the Vrije Universiteit Brussel (IOF Project 4R-ART Nr 2042). Work in G.K.'s laboratory is supported by the UK Biotechnology and Biological Sciences Research Council and Medical Research Council. The authors have no conflicts of interest.

The soluble isoform of human FcɛRI is an endogenous inhibitor of IgE-mediated mast cell responses.

BACKGROUND: The soluble isoform of FcɛRI, the high-affinity IgE receptor (sFcεRI), is a protein of the IgE network with poorly defined functions. OBJECTIVE: To define cellular sources and signals that result in the production of human sFcεRI and study its in vivo functions. METHODS: FcεRI-transfected human cell lines (MelJuso), human monocyte-derived dendritic cells (moDCs), and murine bone marrow-derived mast cells (MC) were stimulated by FcεRI cross-linking and release of sFcεRI was analyzed (ELISA, Western Blot). Lysosomal-associated membrane protein 1 degranulation assays and human basophil activation tests (BATs) were used to study IgE-dependent activation. Recombinant sFcεRI (rsFcεRI) was used to assess its role in murine models of anaphylaxis with WT (wild-type) and IgE-/- (IgE-deficient) mice. RESULTS: Antigen-specific cross-linking of IgE-loaded FcɛRI on MelJuso cells that express the trimeric or tetrameric receptor isoform induced the production of sFcεRI. Using MCs and moDCs, we confirmed that IgE/FcɛRI activation induces sFcɛRI release. We demonstrated that generation of sFcɛRI requires Src phosphorylation and endo/lysosomal acidification. In experimental mouse models, sFcɛRI diminishes the severity of IgE-mediated anaphylaxis. BATs confirmed that, comparable to the anti-IgE monoclonal antibody omalizumab, sFcɛRI is an inhibitor of the human innate IgE effector axis, implying that sFcɛRI and omalizumab potentially inhibit each other in vivo. CONCLUSION: sFcɛRI is produced after antigen-specific IgE/FcɛRI-mediated activation signals and functions as an endogenous inhibitor of IgE loading to FcɛRI and IgE-mediated activation. Our results imply, therefore, that sFcɛRI contributes to a negative regulatory feedback loop that aims at preventing overshooting responses after IgE-mediated immune activation.

Posttransplant monomorphic Burkitt's lymphoma: clinical characteristics and outcome of a multicenter series.

Burkitt's monomorphic posttransplant lymphoproliferative disorder (B-PTLD) is an uncommon subtype of PTLD. Owing to the paucity of this complication, clinical characteristics and outcome has not been fully described. Clinical characteristics and outcomes of 20 patients diagnosed with B-PTLD from 10 transplant centers belonging to the GEL/TAMO group were reviewed. Median time from transplant to B-PTLD was 7.2 years. All the cases fulfill the morphologic and genetic criteria of B-PTLD, whereas Epstein-Barr virus (EBV) was detected in 70% of cases. Patients were treated with different chemotherapy combinations, and three patients received upfront rituximab monotherapy. The great majority of patients receiving CHOP-like regimens attained a complete response (CR) (73%), similar to that obtained with dose-intensive chemotherapy (83% CR). In contrast, patients receiving upfront rituximab monotherapy required subsequent chemotherapy. Two patients (10%) died during treatment due to infection. The median progression-free survival and overall survival (OS) were 16 months and 139 months, respectively. When analyzing variables predicting for OS, we found that patients with bone marrow involvement had an adverse prognosis, with a median OS of 6 months (p = 0.008). In conclusion, B-PTLD is an uncommon complication usually associated with EBV infection and with an aggressive clinical course, particularly in patients with bone marrow involvement. High-dose chemoimmunotherapy obtained similar responses to R-CHOP, suggesting that R-CHOP could be an adequate alternative for these patients. In contrast, rituximab monotherapy does not seem to be effective enough to control the disease.

MPT64 patch test for the diagnosis of active pulmonary tuberculosis: a randomised controlled trial in Peru.

SETTING: There remains a lack of effective and inexpensive diagnostic tools for active tuberculosis (TB) disease. Testing immune responses to proteins secreted by Mycobacterium tuberculosis, such as MPT64, may be a diagnostic option. OBJECTIVE: To evaluate the sensitivity and specificity of a patch test using MPT64 for the diagnosis of active TB disease. DESIGN: This randomised, double-blind, placebo-controlled, prospective study in Lima, Peru, involved 55 healthy controls and 457 symptomatic individuals referred for routine TB testing by the National TB Control Programme. All subjects underwent a comprehensive diagnostic workup, and received an active patch on one arm and a placebo patch on the opposite arm, which were read after 4 days. RESULTS: Eighty-one (18%) of the symptomatic participants were classified as having definite TB, while an additional 98 (21%) had probable TB. The patch tests performed the same in both groups, with a sensitivity of 27% and specificity of 74%. The area under the receiver operating characteristic curve was 0.495 (95%CI 0.425-0.565). CONCLUSIONS: Contrary to existing literature, the MPT64 patch was not sensitive and specific to detect active TB. Given the potential of the test, understanding possible differences in the protein source or underlying genetic factors should be explored further.

[Bronchiectasis]. / Bronquiectasias.

Bronchiectasis is defined as irreversible dilatation of the bronchial lumen. Bronchiectasis not due to cystic fibrosis is the third most common chronic inflammatory disease of the airway, after asthma and chronic obstructive pulmonary disease (CPOD). The pathogenesis of the disease is characterised by chronic dilatation, with irreversible and usually progressive destruction of the bronchial wall as a consequence of the vicious pathogenic circle described by E. Cole, of infection, inflammation, injury to the mucociliary system, and cyclical repair of the airway. The disease can have different causes, and usually presents with cough and chronic expectoration, and repeated, acute, infectious exacerbations. Suspected diagnosis is clinical, and must be confirmed by high resolution computerised tomography. The treatment comprises diverse regimens aimed at controlling infection and bronchial inflammation. Nutritional aspects should be considered as part of treatment, as well as the management of secretions, muscle training and the management of complications and comorbidities.

An improved IVM method for cumulus-oocyte complexes from small follicles in polycystic ovary syndrome patients enhances oocyte competence and embryo yield.

STUDY QUESTION: Are meiotic and developmental competence of human oocytes from small (2-8 mm) antral follicles improved by applying an optimized IVM method involving a prematuration step in presence of C-Type Natriuretic Peptide (CNP) followed by a maturation step in presence of FSH and Amphiregulin (AREG)? SUMMARY ANSWER: A strategy involving prematuration culture (PMC) in the presence of CNP followed by IVM using FSH + AREG increases oocyte maturation potential leading to a higher availability of Day 3 embryos and good-quality blastocysts for single embryo transfer. WHAT IS KNOWN ALREADY: IVM is a minimal-stimulation ART with reduced hormone-related side effects and risks for the patients, but the approach is not widely used because of an efficiency gap compared to conventional ART. In vitro systems that enhance synchronization of nuclear and cytoplasmic maturation before the meiotic trigger are crucial to optimize human IVM systems. However, previous PMC attempts have failed in sustaining cumulus-oocyte connections throughout the culture period, which prohibited a normal cumulus-oocyte communication and precluded an adequate response by the cumulus-oocyte complex (COC) to the meiotic trigger. STUDY DESIGN, SIZE, DURATION: A first prospective study involved sibling oocytes from a group of 15 patients with polycystic ovary syndrome (PCOS) to evaluate effects of a new IVM culture method on oocyte nuclear maturation and their downstream developmental competence. A second prospective study in an additional series of 15 women with polycystic ovaries characterized and fine-tuned the culture conditions. PARTICIPANTS/MATERIALS, SETTING, METHODS: Fifteen women with PCOS (according to Rotterdam criteria) underwent IVM treatment after 3-5 days of highly purified human menopausal gonadotropin (HP-hMG) stimulation and no human chorionic gonadotropin (hCG) trigger before oocyte retrieval. A first study was designed with sibling oocytes to prospectively evaluate the impact of an IVM culture method: 24 h PMC with CNP + 30 h IVM with FSH and AREG, on embryo yield, in comparison to the standard (30 h) IVM clinical protocol (Group I, n = 15). A second prospective study was performed in 15 women with polycystic ovaries, to characterize and optimize the PMC conditions (Group II, n = 15). The latter study involved the evaluation of oocyte meiotic arrest, the preservation of cumulus-oocyte transzonal projections (TZPs), the patterns of oocyte chromatin configuration and cumulus cells apoptosis following the 24 and 46 h PMC. Furthermore, oocyte developmental potential following PMC (24 and 46 h) + IVM was also evaluated. The first 20 good-quality blastocysts from PMC followed by IVM were analysed by next generation sequencing to evaluate their aneuploidy rate. MAIN RESULTS AND THE ROLE OF CHANCE: PMC in presence of CNP followed by IVM using FSH and AREG increased the meiotic maturation rate per COC to 70%, which is significantly higher than routine standard IVM (49%; P ≤ 0.001). Hence, with the new system the proportion of COCs yielding transferable Day 3 embryos and good-quality blastocysts increased compared to routine standard IVM (from 23 to 43%; P ≤ 0.001 and from 8 to 18%; P ≤ 0.01, respectively). CNP was able to prevent meiosis resumption for up to 46 h. After PMC, COCs had preserved cumulus-oocyte TZPs. The blastocysts obtained after PMC + IVM did not show increased aneuploidy rates as compared to blastocysts from conventional ART. LIMITATIONS REASONS FOR CAUTION: The novel IVM approach in PCOS patients was tested in oocytes derived from small antral follicles which have an intrinsically low developmental potential. Validation of the system would be required for COCs from different (larger) follicular sizes, which may involve further adjustment of PMC conditions. Furthermore, considering that this is a novel strategy in human IVM treatment, its global efficiency needs to be confirmed in large prospective randomized controlled trials. The further application in infertile patients without PCOS, e.g. cancer patients, remains to be evaluated. WIDER IMPLICATIONS OF THE FINDINGS: The findings of this pilot study suggest that the efficiency gap between IVM and conventional IVF can be reduced by fine-tuning of the culture methods. This novel strategy opens new perspectives for safe and patient-friendly ART in patients with PCOS. STUDY FUNDING/COMPETING INTEREST(S): IVM research at the Vrije Universiteit Brussel has been supported by grants from: the Institute for the Promotion of Innovation by Science and Technology in Flanders (Agentschap voor Innovatie door Wetenschap en Technologie-IWT, project 110680); the Fund for Research Flanders (Fonds Wetenschappelijk Onderzoek-Vlaanderen-FWO, project G.0343.13), the Belgian Foundation Against Cancer (HOPE project, Dossier C69). The authors have no conflicts of interest.

Características antropométricas maternas y del lactante, correlacionado a la concentración proteica del calostro y leche madura: un estudio longitudinal / Anthropometric characteristics of the mother and the infant, correlated to the protein concentration of colostrum and mature milk: a longitudinal study

Objetivo: Determinar las características antropométricas maternas y del lactante correlacionados a la concentración proteica del calostro y la leche madura en el Hospital "El Carmen", Huancayo. Método: Estudio analítico, longitudinal, correlacional; Muestra: 35 (madres y lactantes), quienes cumplían criterios de selección. Instrumento: ficha de recolección de datos consignando las medidas antropométricas maternas, del lactante y resultados de concentración proteica del calostro y leche madura; medición de variables en dos fases (Al parto y cuarto mes post- parto). Se utilizó correlación R de Spearman, Pearson y regresión logística multivariada. Resultados: La mediana de edad de las madres fue de 20 años, el IMC promedio pre- gestacional fue 23,55, mediana pre- parto fue de 26,71, y cuarto mes post- parto fue de 24,42; En los lactantes el sexo predominante fue el femenino 54,29%, la mediana del peso al nacer fue 3.030 kg y a los 4 meses tuvieron un peso promedio de 6,580 kg. El análisis bivariado mostró significancia estadística entre la concentración de proteínas del calostro y peso al cuarto mes (p=0.0119) y la concentración de proteínas de leche madura con respecto a la talla del lactante al cuarto mes (p=0.0041). El análisis multivariado para el peso del recién nacido, tiene relación con el Índice de Masa Corporal Pre- gestacional (p<0.011), R2 :0.481. Conclusión: Las características antropométricas de la madre y del lactante están correlacionados a la concentración proteica del calostro y leche madura Palabras clave: Índice de masa corporal, Recién nacido, Peso al nacer, Leche Materna, Proteínas.

Objective: To determine maternal and infant anthropometric characteristics correlated to protein concentration of colostrum and mature milk in the Hospital "El Carmen", Huancayo Method: analytical, correlational longitudinal study; Shows: 35 (mothers and infants) who met the selection criteria. Instrument: data collection sheet consigning maternal anthropometric measurements, infant and results of protein concentration of colostrum and mature milk; measurement variables in two phases (Al childbirth and fourth month postpartum). Spearman correlation R, Pearson and multivariate logistic regression was used. Results: The median age of the mothers was 20 years, mean BMI was pre- gestational 23.55, median pre- delivery was 26.71, and fourth month postpartum was 24.42; In infants the predominant female sex was 54.29%, the median birth weight was 3,030 kg and at 4 months had an average weight of 6,580 kg. The bivariate analysis showed statistical significance between protein concentration of colostrum and fourth month weight (p = 0.0119) and the protein concentration of mature milk with respect to the size of the infant to four months (p = 0.0041). Multivariate for birth weight, analysis is related to the gestational Body Mass Index Pre (p <0.011), R2: 0.481. Conclusion: The anthropometric characteristics of the mother and infant are correlated to protein concentration of colostrum and mature milk Keywords: Body mass index, Newborn, Birth weight, Breast Milk, Proteins.

Rewiring of the apoptotic TGF-ß-SMAD/NFκB pathway through an oncogenic function of p27 in human papillary thyroid cancer.

Papillary thyroid carcinoma (PTC), the most frequent thyroid cancer, is characterized by low proliferation but no apoptosis, presenting frequent lymph-node metastasis. Papillary thyroid carcinoma overexpress transforming growth factor-beta (TGF-ß). In human cells, TGF-ß has two opposing actions: antitumoral through pro-apoptotic and cytostatic activities, and pro-tumoral promoting growth and metastasis. The switch converting TGF-ß from a tumor-suppressor to tumor-promoter has not been identified. In the current study, we have quantified a parallel upregulation of TGF-ß and nuclear p27, a CDK2 inhibitor, in samples from PTC. We established primary cultures from follicular epithelium in human homeostatic conditions (h7H medium). TGF-ß-dependent cytostasis occurred in normal and cancer cells through p15/CDKN2B induction. However, TGF-ß induced apoptosis in normal and benign but not in carcinoma cultures. In normal thyroid cells, TGF-ß/SMAD repressed the p27/CDKN1B gene, activating CDK2-dependent SMAD3 phosphorylation to induce p50 NFκB-dependent BAX upregulation and apoptosis. In thyroid cancer cells, oncogene activation prevented TGF-ß/SMAD-dependent p27 repression, and CDK2/SMAD3 phosphorylation, leading to p65 NFκB upregulation which repressed BAX, induced cyclin D1 and promoted TGF-ß-dependent growth. In PTC samples from patients, upregulation of TGF-ß, p27, p65 and cyclin D1 mRNA were significantly correlated, while the expression of the isoform BAX-ß, exclusively transcribed in apoptotic cells, was negatively correlated. Additionally, combined ERK and p65 NFκB inhibitors reduced p27 expression and potentiated apoptosis in thyroid cancer cells while not affecting survival in normal thyroid cells. Our results therefore suggest that the oncoprotein p27 reorganizes the effects of TGF-ß in thyroid cancer, explaining the slow proliferation but lack of apoptosis and metastatic behavior of PTC.

Educación médica para la Atención Primaria de Salud: visión de los docentes y estudiantes / Perceptions of faculty members and students about undergraduate training in primary health care

BACKGROUND: It is unknown if medical education is preparing physicians to successfully work at primary health care settings. AIM: To explore what are the perceptions of faculty members and students about the type of physician needed and if medical education is coherent with the practice of primary health care. MATERIAL AND METHODS: Fifteen semi-structured interviews to key informants from faculty members and ten focus groups with students were carried out. RESULTS: Important influences of role modelling and hidden curriculum were found, especially in relation to the type of physician needed, generalist or specialist, and in relation to the places where the clinical practices were done. Although primary health care was declared in the profiles, most of clinical practices were done at hospitals and supervised by specialists. Working at primary health care is seen as a temporary work, not valued by professionals nor by the society. CONCLUSIONS: Medical Schools are not preparing professionals for primary health care but for hospital care and specialized medicine.

Factores de riesgo asociados a hígado graso de origen no alcohólico en una población del Caribe Colombiano / Risk Factors Associated with Nonalcoholic Fatty Liver Disease in a Colombian Caribbean Population

La enfermedad hepática grasa no alcohólica NAFLD (Non-alcoholic fatty liver disease) es un problema de salud, así lo demuestran las diversas investigaciones alrededor del mundo, donde se evidencia una alta prevalencia de la patología. El objetivo del estudio piloto fue describir en términos de variables demográficas, epidemiológicas y bioquímicas la relación de factores de riesgo asociados a NAFLD de una muestra de pacientes que acudieron a la sección de Hígado y vías biliares del Hospital Universitario del Caribe (Cartagena, Colombia). El estudio piloto fue descriptivo prospectivo, fueron revisadas historias clínicas para análisis de variables epidemiológicas-demográficas y valorados por medición espectrofotométrica la actividad enzimática de transaminasas para la relación De Ritis (AST/ALT), niveles de colesterol y triglicéridos. Los resultados sugieren que la población subdividida en obesos, diabetes mellitus, hipertensos/obesos, diabetes/obesos y principalmente del género femenino presenta mayor riesgo de padecer hígado graso, con lo que amerita un estudio bioquímico molecular y un diagnóstico definitivo de NAFLD en la población de estudio.

Nonalcoholic fatty liver disease (NAFLD) is a health problem that has been demonstrated by research from everywhere around the world where the disease's prevalence appears to be high. The aim of this pilot study was to describe the relationship of risk factors associated with NAFLD in terms of demographic, epidemiological and biochemical variables. Patients included in the study were drawn from people who came to the liver and biliary tract section of the Hospital Universitario del Caribe in Cartagena, Colombia. This was a prospective and descriptive pilot study that reviewed medical records for analysis of epidemiological and demographic variables. It assessed spectrophotometric measurement of transaminases for determination of De Ritis ratios (AST/ALT), and relationship to levels of cholesterol and triglycerides. The results suggest that risks of developing NAFLD were higher among women, among patients who were obese, among patients who had diabetes mellitus, among patients who had hypertension and obesity, and among patients who had diabetes combined with obesity. Patients at risk merit biochemical molecular study and a definitive diagnosis.

Serology and longevity of immunity against Echinococcus granulosus in sheep and llama induced by an oil-based EG95 vaccine.

An oil-based formulation of the EG95 vaccine to protect grazing animals against infection with Echinococcus granulosus was formulated in Argentina. The efficacy of the vaccine was monitored by serology in sheep and llama (Lama glama) and was compared to the serology in sheep previously published using a QuilA-adjuvanted vaccine. Long-term efficacy was also tested in sheep by challenging with E. granulosus eggs of the G1 strain 4 years after the beginning of the trial. The serological results for both sheep and llama were similar to those described previously, except that there was a more rapid response after the first vaccination. A third vaccination given after 1 year resulted in a transient boost in serology that lasted for about 12 months, which was similar to results previously described. Sheep challenged after 4 years with three vaccinations presented 84·2% reduction of live cysts counts compared with control group, and after a fourth vaccination prior to challenge, this reduction was 94·7%. The oil-based vaccine appeared to be bio-equivalent to the QuilA vaccine.

[Sociodemographic characteristics and geographic variability associated with patient satisfaction in Primary Care]. / Características sociodemográficas y variabilidad geográfica relacionada con la satisfacción del paciente en Atención Primaria.

BACKGROUND: Patient satisfaction with health services plays an important role in programs to improve the quality of care from the point of view of users. The objectives of this study were: To identify sociodemographic characteristics associated with patient satisfaction in the care provided by family doctors in Primary Health Care (PHC) centres, and describe the geographical variability of this phenomenon in the Spanish National Health Service. MATERIAL AND METHODS: The data come from the European Health Survey (2009). Prevalence ratios (crude and adjusted) of the characteristics associated with both excellent satisfaction and dissatisfaction using Poisson regression, and their geographical variability are discussed. RESULTS: About one in every 3 users of the PHC believes that the care provided was excellent, while 6.7% were dissatisfied. There is a wide variability in the perception of satisfaction among the various regional health services, with prevalence ranging between 10.9% and 55.2%. Moreover, this assessment is closely related to age, level of self-perceived health, mental health, previous hospitalisation, chronic disease status, and limitations in daily activities. CONCLUSIONS: Satisfaction with the care provided by the PHC physician is relatively high. However, the distribution between regions and socio-demographic characteristics and health status of the user is heterogeneous.

Human cumulus-enclosed germinal vesicle oocytes from early antral follicles reveal heterogeneous cellular and molecular features associated with in vitro maturation capacity.

STUDY QUESTION: Are oocyte size, chromatin remodelling, transcriptional activity and mitochondrial distribution in human immature oocytes from early antral follicles retrieved for in vitro maturation (IVM) associated with the acquisition of meiotic competence? SUMMARY ANSWER: Oocyte size, chromatin compaction, cessation of RNA synthesis and mitochondria rearrangement around the nucleus are associated with the oocyte's potential to resume meiosis in vitro. WHAT IS KNOWN ALREADY: Information on oocyte features that confer meiotic competence in human mainly derives from germinal vesicle (GV) oocytes that failed to resume meiosis following an hCG trigger after ovulation induction cycles. Characterization of cumulus-enclosed GV oocytes from small antral follicles prior to IVM provides knowledge on the initial oocyte status and suggests culture requirements in order to promote oocyte competence in vitro. STUDY DESIGN, SIZE, DURATION: Prospective collection of 107 oocytes immediately after retrieval (before IVM) and of 293 GV oocytes that had failed to resume meiosis (after IVM). PARTICIPANTS/MATERIALS, SETTING, METHODS: Human oocytes were collected from women with polycystic ovary syndrome (PCOS), receiving in total 450 IU of highly purified-hMG for IVM treatment (patients) or who donated oocytes for IVM research (donors). Oocytes at GV-stage were retrieved from follicles <10 mm (range 2-10 mm) diameter, before IVM (oocytes at retrieval) or those that failed to mature after IVM (meiotically incompetent). Oocytes were allocated for either mitochondrial staining, by incubating in mitotracker red and then fixed; or for nascent RNA staining, which was assessed by fluorescent labelling (Click-iT(®) RNA Assay). In every case, oocyte diameter was recorded and chromatin was stained after oocyte fixation. GV-stage oocytes were analysed by confocal laser-scanning microscopy and their characteristics were compared and related to their meiotic competence. MAIN RESULTS AND THE ROLE OF CHANCE: Analysis of oocytes at the immature GV-stage revealed that oocytes at retrieval were significantly larger than those that failed to resume meiosis after IVM (112.7 versus 109.6 µm, P < 0.0001). Oocytes assessed at retrieval showed that 50.6% had a condensed chromatin configuration (perinucleolar chromatin rim) and were consistently transcriptionally silent. This rate matched maturation rates in our current in vitro culture system (49%). However, oocytes that had not reinitiated meiosis after 30 h IVM demonstrated, apart of being smaller in diameter, significantly higher rates of dispersed or intermediate chromatin (P = 0.005). Analysis of mitochondrial distribution revealed that many oocytes at retrieval displayed mitochondrial internalization towards the nucleus (12/30) or a perinuclear mitochondrial distribution (6/30). These mitochondrial patterns were observed more rarely in GV incompetent oocytes following 30 h IVM (16/98 and 1/98, respectively). LIMITATIONS, REASONS FOR CAUTION: Most of the analyses involved the use of invasive techniques. Hence, despite the fact that these data deliver essential information on the intrinsic oocyte maturational and developmental status, a direct match with embryological outcomes could not be established. WIDER IMPLICATIONS OF THE FINDINGS: The evidence described here can aid in tailoring IVM systems in order to promote completion of nuclear and cytoplasmic maturation of unexpanded cumulus-oocyte complexes. STUDY FUNDING/COMPETING INTERESTS: This study was supported by research grants by the Institute for the Promotion of Innovation by Science and Technology in Flanders, project numbers IWT 130327 and 110680; the Fund for Research Flanders, project number FWO G.0343.13, the Belgian Foundation Against Cancer (HOPE project) and COOK Medical. None of the authors has any competing interest to declare.

Human in vitro oocyte maturation is not associated with increased imprinting error rates at LIT1, SNRPN, PEG3 and GTL2.

STUDY QUESTION: Does in vitro maturation (IVM) of cumulus-enclosed germinal vesicle (GV) stage oocytes retrieved from small antral follicles in minimally stimulated cycles without an ovulatory hCG dose induce imprinting errors at LIT1, SNRPN, PEG3 and GTL2 in human oocytes? SUMMARY ANSWER: There is no significant increase in imprinting mutations at LIT1, SNRPN, PEG3 and GTL2 after IVM of cumulus-enclosed GV oocytes from small antral follicles in minimally stimulated cycles without hCG priming. WHAT IS KNOWN ALREADY: Animal models have generally demonstrated correct methylation imprint establishment for in vitro grown and matured oocytes. For human IVM, well-designed studies allowing conclusions on imprint establishment are currently not available. STUDY DESIGN, SIZE, DURATION: Immature oocyte-cumulus complexes from 2 to 9 mm follicles were retrieved in polycystic ovary syndrome (PCOS) subjects in minimally stimulated cycles without hCG priming and matured in vitro. In vivo grown oocytes were retrieved after conventional ovarian stimulation for IVF/ICSI or after ovulation induction. Imprinting error rates at three maternally methylated (LIT1, SNRPN and PEG3) and one paternally methylated (GTL2) imprinted genes were compared in 71 in vitro and 38 in vivo matured oocytes. PARTICIPANTS/MATERIALS, SETTING, METHODS: The limiting dilution bisulfite sequencing technique was applied, allowing increased sensitivity based on multiplex PCR for the imprinted genes and the inclusion of non-imprinted marker genes for cumulus cell DNA contamination. MAIN RESULTS AND THE ROLE OF CHANCE: In vitro as well as in vivo matured oocytes showed only a few abnormal alleles, consistent with epimutations. The abnormalities were more frequent in immature than in mature oocytes for both groups, although no significant difference was reached. There was no statistically significant increase in imprinting errors in IVM oocytes. LIMITATIONS, REASONS FOR CAUTION: This single cell methylation analysis was restricted to a number of well-selected imprinted genes. Genome-wide methylation analysis of single human oocytes is currently not possible. WIDER IMPLICATIONS OF THE FINDINGS: IVM is a patient-friendly alternative to conventional ovarian stimulation in PCOS patients and is associated with reduced gonadotrophin costs and avoidance of OHSS. The results of this study show for the first time that optimized human IVM procedures have no significant effects on the establishment of maternal DNA methylation patterns at LIT1, SNRPN, PEG3 and GTL2. STUDY FUNDING/COMPETING INTERESTS: This study was supported by research funds from Agentschap voor Innovatie door Wetenschap en Technologie (IWT-TBM 110680), Wetenschappelijk Fonds Willy Gepts (WFWG 2011) and German Research Foundation (HA 1374/12-2). There are no competing interests.

Effect of energy sources on the apparent total tract digestibility and excretion of nutrients by bovine cattle / Efecto de fuentes energéticas sobre la digestibilidad aparente total y excreción de nutrientes por bovinos

Objective. To evaluate the effect of three energy sources on the intake, total apparent digestibility and excretion of nutrients in cattle diet. Materials and methods. Six ruminally cannulated cows (730 ± 70 kg) were distributed into three treatments in a replicated 3x3 Latin square experimental design, where: (I) Control: Low ether extract diet (3.50% EE); (II) Soybean: High ether extract diet (5.30% EE) with inclusion of 15% soybean and (III) Citrus pulp: Low ether extract diet (3.00% EE) and high pectin involvement with inclusion of 15% citrus pulp. To determine the digestibility of DM and its fractions, chromic oxide was used as a marker. Nutrient excretion was calculated from the digestibility coefficient of each fraction. Results. The soybean treatment reduced (p<0.05) dry matter, crude protein, nitrogen-free extract and organic matter intake and increased ether extract intake. There was no effect (p>0.05) of the energy source on the digestibility coefficients of DM, CP, NDF, EE, NFE or OM. The TDN value was higher for the soybean treatment. The excretion of DM, NFE and OM was lower (p<0.05) for the soybean treatment. All energy sources influenced the excretion of crude protein. Conclusions. The energy sources used did not affect the digestibility of the diets and are indicated as high potential sources to be used in cattle.

Objetivo. Evaluar el efecto de tres fuentes energéticas sobre el consumo, la digestibilidad aparente total y la excreción de nutrientes de la dieta en bovinos. Materiales y métodos. Seis vacas (730±70 kg) canuladas en rumen fueron distribuidas en tres tratamientos, en un diseño experimental cuadrado latino 3x3 replicado, a saber: (I) Control: Dieta de bajo extracto etéreo (3.50% de EE); (II) Soya: Dieta de alto extracto etéreo (5.30% de EE) con inclusión de 15% de grano de soya y (III) Pulpa Cítrica: Dieta de bajo extracto etéreo (3.00% de EE) y alta participación de pectina con inclusión de 15% de pulpa cítrica. Para determinar la digestibilidad de la MS y sus fracciones, fue utilizado el marcador de óxido de cromo. La excreción de nutrientes, fue calculada a partir de los coeficientes de digestibilidad de cada fracción. Resultados. El tratamiento con soya redujo (p<0.05) el consumo de materia seca, proteína bruta, extracto no nitrogenado y materia orgánica, de igual manera, incrementó el consumo de extracto etéreo. No no se observó efecto (p˃0.05) de fuente energética para los coeficientes de digestibilidad de la MS, PB, FDN, EE, ENN y MO. El valor de NDT fue mayor para el tratamiento con soya. La excreción de MS, ENN y MO fue menor (p<0.05) para el tratamiento con soya. Todas las fuentes energéticas influyeron en la excreción de proteína bruta. Conclusiones. Las fuentes energéticas utilizadas, no afectaron la digestibilidad de las dietas, y estan indicadas como fuentes de elevado potencial en la alimentación de bovinos.

Amblyopia secondary to iris cyst.

CLINICAL CASE: A 5 year-old child diagnosed with moderate anisometropic amblyopia secondary to primary cyst of iris pigment epithelium. He was evaluated with ultrasound biomicroscopy (BMU) and optical coherence tomography (OCT) of anterior segment. DISCUSSION: The OCT, although with some limitations, is a useful tool to study the anterior segment. It is probably more recommendable than BMU in the childhood.

Cocoa intake attenuates oxidative stress associated with rat adjuvant arthritis.

Cocoa contains flavonoids with antioxidant properties. The aim of this study was to ascertain the effect of cocoa intake on oxidative stress associated with a model of chronic inflammation such as adjuvant arthritis. Female Wistar rats were fed with a 5% or 10% cocoa-enriched diet or were given p.o. a quercetin suspension every other day for 10 days. Arthritis was induced by a heat-killed Mycobacterium butyricum suspension. Reactive oxygen species (ROS) produced by macrophages, and splenic superoxide dismutase (total, cytoplasmic and mitochondrial) and catalase activities were determined. Clinically, joint swelling in arthritic rats was not reduced by antioxidants; however, the 5% cocoa diet and quercetin administration reduced ROS production. Moreover, the 5% cocoa diet normalized the activities of superoxide dismutase and catalase. In conclusion, a cocoa diet reduces the oxidative stress associated with a chronic inflammatory pathology, although it was not enough to attenuate joint swelling.