Cardiosome-mediated protection in myocardial ischemia.

Cardiosomes, exosomes released in cardiospheres by cardiomyocytes and progenitor cells, communicate locally and at a distance from different tissues, promoting beneficial cellular changes. For example, miRNAs have emerged as regulators of intercellular communication via transport by extracellular vesicles in general and cardiosomes specifically. Although cardiosomes are considered biomarkers owing to their immense biomedical application in various clinical fields, their role in cardiovascular diseases remains unclear. This mini-review examines the experimental and clinical evidence for cardiosomes as non-invasive diagnostic, treatment and prognostic tools in acute myocardial infarction, the novelty of which is often lost in medical practice. In addition, we discuss the potential role of cardiosomes in physiologic mechanisms and cell signaling in cardiac conditioning strategies against reperfusion injury.

Actin-Cytoskeleton Drives Caveolae Signaling to Mitochondria during Postconditioning.

Caveolae-associated signaling toward mitochondria contributes to the cardioprotective mechanisms against ischemia-reperfusion (I/R) injury induced by ischemic postconditioning. In this work, we evaluated the role that the actin-cytoskeleton network exerts on caveolae-mitochondria communication during postconditioning. Isolated rat hearts subjected to I/R and to postconditioning were treated with latrunculin A, a cytoskeleton disruptor. Cardiac function was compared between these hearts and those exposed only to I/R and to the cardioprotective maneuver. Caveolae and mitochondria structures were determined by electron microscopy and maintenance of the actin-cytoskeleton was evaluated by phalloidin staining. Caveolin-3 and other putative caveolae-conforming proteins were detected by immunoblot analysis. Co-expression of caveolin-3 and actin was evaluated both in lipid raft fractions and in heart tissue from the different groups. Mitochondrial function was assessed by respirometry and correlated with cholesterol levels. Treatment with latrunculin A abolishes the cardioprotective postconditioning effect, inducing morphological and structural changes in cardiac tissue, reducing F-actin staining and diminishing caveolae formation. Latrunculin A administration to post-conditioned hearts decreases the interaction between caveolae-forming proteins, the co-localization of caveolin with actin and inhibits oxygen consumption rates in both subsarcolemmal and interfibrillar mitochondria. We conclude that actin-cytoskeleton drives caveolae signaling to mitochondria during postconditioning, supporting their functional integrity and contributing to cardiac adaption against reperfusion injury.

The Development of a Virtual World Problem-Based Learning Tutorial and Comparison With Interactive Text-Based Tutorials.

Collaborative learning through case-based or problem-based learning (PBL) scenarios is an excellent way to acquire and develop workplace knowledge associated with specific competencies. At St George's, University of London we developed an interactive online form of decision-based PBL (D-PBL) for our undergraduate medical course using web-based virtual patients (VPs). This method of delivery allowed students to consider options for clinical management, to take decisions and to explore the consequences of their chosen actions. Students had identified this as a more engaging type of learning activity compared to conventional paper-based/linear PBL and demonstrated improved exam performance in controlled trials. We explored the use of Second Life (SL), a virtual world and immersive 3D environment, as a tool to provide greater realism than our interactive image and text-based D-PBL patient cases. Eighteen separate tutorial groups were provided with their own experience of the same patient scenario in separate locations within the virtual world. The study found that whilst a minority of students reported that the Second Life experience felt more realistic, most did not. Students favored the simpler interaction of the web-based VPs, which already provided them with the essential learning needed for practice. This was in part due to the time proximity to exams and the extra effort required to learn the virtual world interface. Nevertheless, this study points the way towards a scalable process for running separate PBL sessions in 3D environments.

Pioneer plant species and fungal root endophytes in metal-polluted tailings deposited near human populations and agricultural areas in Northern Mexico.

As a consequence of industrial mining activity, high volumes of tailings are scattered around Mexico. Frequently, tailings contain heavy metals (HM) which entail threats against all organisms. The aim of this research was to identify plants and root fungal endophytes in polymetallic polluted tailings with the potential to be used in strategies of bioremediation. Four deposits of mine wastes, situated in a semi-arid region near urban and semi-urban populations, and agricultural areas, were studied. The physical and chemical characteristics of substrates, accumulation of HM in plant tissues, root colonization between arbuscular mycorrizal (AMF) and dark septate endophyte (DSE) fungi, and the identification of DSE fungi isolated from the roots of two plant species were studied. Substrates from all four sites exhibited extreme conditions: high levels in sand; low water retention; poor levels in available phosphorus and nitrogen content; and potentially toxic levels of lead (Pb), cadmium (Cd), and zinc (Zn). The native plants Lupinus campestris, Tagetes lunulata, and Cerdia congestiflora, as well as the exotic Cortaderia selloana and Asphodelus fistulosus, demonstrated a relevant potential role in the phytostabilization and/or phytoextraction of Pb, Cd, and Zn, according to the accumulation of metal in roots and translocation to shoots. Roots of eleven analyzed plant species were differentially co-colonized between AMF and DSE fungi; the presence of arbuscules and microsclerotia suggested an active physiological interaction. Fourteen DSE fungi were isolated from the inner area of roots of T. lunulata and Pennisetum villosum; molecular identification revealed the predominance of Alternaria and other Pleosporales. The use of native DSE fungi could reinforce the establishment of plants for biological reclamation of mine waste in semi-arid climate. Efforts are needed in order to accelerate a vegetation practice of mine wastes under study, which can reduce, in turn, their potential ecotoxicological impact on organisms, human populations, and agricultural areas.

Relevance of endoplasmic reticulum and mitochondria interactions in age-associated diseases.

Although the elixir of youth remains in the darkness, medical and scientific advances have succeeded in increasing human longevity; however, the predisposition to disease and its high economic cost are raising. Different strategies (e.g., antioxidants) and signaling pathways (e.g., Nrf2) have been identified to help regulate disease progression, nevertheless, there are still missing links that we need to understand. Contact sites called mitochondria-associated membranes (MAM) allow bi-directional communication between organelles as part of the essential functions in the cell to maintain its homeostasis. Different groups have deeply studied the role of MAM in aging; however, it's necessary to analyze their involvement in the progression of age-related diseases. In this review, we highlight the role of contact sites in these conditions, as well as the morphological and functional changes of mitochondria and ER in aging. We emphasize the intimate relationship between both organelles as a reflection of the biological processes that take place in the cell to try to regulate the deterioration characteristic of the aging process; proposing MAM as a potential target to help limit the disease progression with age.

COPe-support - a multi-component digital intervention for family carers for people affected by psychosis: study protocol for a randomized controlled trial.

BACKGROUND: Psychosis often causes significant distress and impacts not only in the individuals, but also those close to them. Many relatives and friends ('carers') provide long-term support and need resources to assist them. We have co-produced a digital mental health intervention called COPe-support (Carers fOr People with Psychosis e-support) to provide carers with flexible access to high quality psychoeducation and interactive support from experts and peers. This study evaluates the effectiveness of COPe-support to promote mental wellbeing and caregiving experiences in carers. METHODS: This study is a single-blind, parallel arm, individually randomized controlled trial (RCT) comparing COPe-support, with attention control. Both groups continue to receive usual care. COPe-support provides interactive web-based psychoeducation on psychosis-related issues, wellbeing-promotion and network support through forums. The attention-control is a non-interactive online information resource pack. Carers living in England are eligible if they provide at least weekly support to a family member or close friend affected by psychosis, and use internet communication (including emails) daily. All trial procedures are run online, including collection of outcome measurements which participants will directly input into our secure platform. Following baseline assessment, a web-based randomization system will be used to allocate 360 carers to either arm. Participants have unlimited access to the allocated condition for 40 weeks. Data collection is at three time points (10, 20, and 40 weeks after randomization). Analyses will be conducted by trial statisticians blinded to allocation. The primary outcome is mental wellbeing measured by Warwick Edinburgh Mental Wellbeing Scale (WEMWBS), at 20 weeks. As well as an intention-to-treat analysis, a complier average causal effect (CACE) analysis will be conducted to estimate the intervention effect in participants who have accessed COPe-support content twice or more. The secondary objectives and analysis will examine other health and caregiving-related outcomes and explore mechanisms. In a process evaluation, we will interview 20% of the intervention arm participants regarding the acceptability of COPe-support. We will explore in detail participants' usage patterns. DISCUSSION: The results of this trial will provide valuable information about the effectiveness of COPe-support in promoting wellbeing and caregiving experiences in carers. TRIAL REGISTRATION: The RCT is registered with the Current Controlled Trials registration (ISRCTN 89563420, registration date: 02/03/2018).

Relevance of Membrane Contact Sites in Cancer Progression.

Membrane contact sites (MCS) are typically defined as areas of proximity between heterologous or homologous membranes characterized by specific proteins. The study of MCS is considered as an emergent field that shows how crucial organelle interactions are in cell physiology. MCS regulate a myriad of physiological processes such as apoptosis, calcium, and lipid signaling, just to name a few. The membranal interactions between the endoplasmic reticulum (ER)-mitochondria, the ER-plasma membrane, and the vesicular traffic have received special attention in recent years, particularly in cancer research, in which it has been proposed that MCS regulate tumor metabolism and fate, contributing to their progression. However, as the therapeutic or diagnostic potential of MCS has not been fully revisited, in this review, we provide recent information on MCS relevance on calcium and lipid signaling in cancer cells and on its role in tumor progression. We also describe some proteins associated with MCS, like CERT, STIM1, VDAC, and Orai, that impact on cancer progression and that could be a possible diagnostic marker. Overall, these information might contribute to the understanding of the complex biology of cancer cells.

Usability evaluation of an eHealth intervention for family carers of individuals affected by psychosis: A mixed-method study.

BACKGROUND: Existing research suggests that eHealth interventions targeting family carers of individuals with long-term illness offer a promising approach to care delivery. In particular, digital psychoeducational interventions with interactive psychosocial support are well-received with high rates of satisfaction and acceptability. However, development of such interventions for psychosis carers is lacking. We developed a multi-component eHealth intervention specifically for carers of individuals affected by psychosis, called COPe-support (Carers fOr People with Psychosis e-support). OBJECTIVE: Using mixed methods to evaluate usability, system heuristics and perceived acceptability, we conducted a usability study to establish the suitability of the intervention prototype for the target user group. METHODS: Twenty-three carers were recruited to the study and participated in a think-aloud test or a remote online trial of the intervention. Qualitative feedback, post-use System Usability Scale (SUS) scores, and real-world usage data collected from the tests were analysed. These were also supplemented with heuristic evaluation data provided by an independent eLearning technology expert. RESULTS: Participants evaluated the intervention content as useful and helpful, and indicated that the system had satisfactory usability with a mean SUS score of 73%, above the usability quality benchmark threshold. Study results identified some minor usability issues, which were corroborated with the eLearning expert's heuristic evaluation findings. We used these results to refine the COPe-support intervention. CONCLUSIONS: The usability study with end-users and service providers identified real-life usage and usability issues. The study results helped us refine COPe-support and its delivery strategy before its launch as part of a large-scale clinical trial.

A Multicomponent eHealth Intervention for Family Carers for People Affected by Psychosis: A Coproduced Design and Build Study.

BACKGROUND: Psychosis, including schizophrenia, is the most common severe mental illness affecting 1% of the population worldwide. A large number of people provide long-term support and care for a relative with psychosis. Although psychoeducational interventions, especially those delivered through a face-to-face group format, have an established evidence base for improving the caregiving experience, well-being, and health outcomes, large-scale implementation and access remain limited. There is a demand for such provision to be made through the internet for greater flexibility and wider access. OBJECTIVE: This study aimed to integrate participatory research methodologies by the public, patients, and carers into the eHealth (electronic health) intervention design and build process to improve the product's usability and acceptability. METHODS: We adapted a structured eHealth intervention build method to include participatory research activities involving key stakeholders and end users to co-design and coproduce our intervention. An expert advisory group (EAG) comprising public involvement members led the formative design and build work using an agile build process. Carers independent from the study were consulted on the evolving drafts of the intervention prototype through focus group meetings. These results were fed back into the intervention build work continuously to ensure end users' input inform every stage of the process. RESULTS: An EAG comprising individuals with lived experience of psychosis, carers, health care professionals, researchers, voluntary organization workers, and eLearning experts (n=14) was established. A total of 4 coproduction workshops were held over 1 year during which the alpha and beta prototypes were designed and built through the participatory research work. Alongside this, 2 rounds of focus group study with carers (n=24, in 4 groups) were conducted to seek consultation on end users' views and ideas to optimize the intervention design and usability. Finally, the EAG carried out a Web-based walk-through exercise on the intervention prototype and further refined it to make it ready for an online usability test. The final product contains multiple sections providing information on psychosis and related caregiving topics and interactive discussion forums with experts and peers for psychosocial support. It provides psychoeducation and psychosocial support for carers through the internet, promoting flexible access and individualized choices of information and support. CONCLUSIONS: The participatory research work led to the coproduction of a eHealth intervention called COPe-support (Carers fOr People with Psychosis e-support). We believe the study methodology, results, and output have optimized the intervention design and usability, fitting the end users' needs and usage pattern. COPe-support is currently being tested for its effectiveness in promoting carers' health outcome through an online randomized controlled trial. TRIAL REGISTRATION: ISRCTN Registry ISRCTN89563420; http://www.isrctn.com/ISRCTN89563420.

Lifestyle patterns and endocrine, metabolic, and immunological biomarkers in European adolescents: The HELENA study.

OBJECTIVE: To evaluate the association of lifestyle patterns related to physical activity (PA), sedentariness, and sleep with endocrine, metabolic, and immunological health biomarkers in European adolescents. METHODS: The present cross-sectional study comprised 3528 adolescents (1845 girls) (12.5-17.5 years) enrolled in the Healthy Lifestyle in Europe by Nutrition in Adolescence Study. Cluster analysis was performed by including body composition, PA by accelerometry, self-reported sedentary behaviors, and sleep duration. We also measured endocrine, metabolic, and immunological biomarkers. RESULTS: Three-cluster solutions were identified: (a) light-PA time, moderate-vigorous-PA time and sedentary time, (b) light-PA time, moderate-vigorous-PA time, sedentary time and sleep time, (c) light-PA time, moderate-vigorous-PA time, sedentary time and body composition. In addition, each cluster solution was defined as: "healthy," "medium healthy," and "unhealthy" according to the presented rating. Analysis of variance showed that overall the healthiest groups from the three clusters analyzed presented a better metabolic profile. A decision tree analysis showed that leptin had a strong association with cluster 3 in both boys and girls, high-density lipoprotein cholesterol had the strongest association with clusters 1 and 3 in boys. Cortisol had the strongest association with cluster 1. HOMA index (homeostatic model assessment) and C3 showed a strong association with cluster 3 in girls. CONCLUSIONS: Our results support the existence of different interactions between metabolic health and lifestyle patterns related to PA, sedentariness, and sleep, with some gender-specific findings. These results highlight the importance to consider multiple lifestyle-related health factors in the assessment of adolescents' health to plan favorable strategies.

Erratum to: Evaluation of Lactobacillus coryniformis CECT5711 strain as a coadjuvant in a vaccination process: a randomised clinical trial in healthy adults.

[This corrects the article DOI: 10.1186/s12986-017-0187-1.].

Erratum to: Evaluation of Lactobacillus coryniformis CECT5711 strain as a coadjuvant in a vaccination process: a randomised clinical trial in healthy adults.

[This corrects the article DOI: 10.1186/s12986-016-0154-2.].

Evaluation of Lactobacillus coryniformis CECT5711 strain as a coadjuvant in a vaccination process: a randomised clinical trial in healthy adults.

BACKGROUND: Although the effects of probiotics on the immune system have been extensively evaluated under disease states, their role in healthy situations remains unclear, since changes are hardly expected under immunological homeostasis. EFSA indicates that vaccination protocols could be used to evaluate the potential role of probiotics to improve the immune response against antigen challenges. The aim of the study was to evaluate the effect of Lactobacillus coryniformis CECT5711 (Lc) on the specific immunity of healthy volunteers undergoing vaccination with Hepatitis A virus (HAV). METHODS: One hundred twenty-three healthy adults were randomised into three groups to follow a 6-week (wk) intervention and all received an intramuscular HAV vaccine 2 weeks after starting the intervention: 1) PRO1 received Lc for 2weeks (1 capsule/day; 3 × 109 CFU/capsule) and placebo capsules after vaccination; 2) PRO2 received a daily capsule of Lc (3 × 109 cfu/day) before and after the challenge; 3) Control group (C) received a daily placebo capsule before and after the vaccine. Blood samples were collected at the beginning (visit 1; V1) and after 2 (V2) and 6 weeks (V3) of the intervention. At each visit, lymphocyte subset counts and cytokine levels were analysed. Specific HAV antibodies were analysed at V1 and V3. To evaluate differences between groups, one-way ANOVA with Bonferroni post-hoc test were used regarding lymphocyte subset counts and specific HAV antibodies production, and Friedman test of related samples and Kendall concordance coefficient for cytokines production. Chi square test was used to analyse seroconversion rates. RESULTS: Specific HAV antibodies were significantly higher in PRO1 (50.54 ± 29.57) compared to C (36.23 ± 16.45) (P = 0.017) and showed an intermediate value in PRO2 (41.61 ± 15.74). Seroconversion rates were similar in the three groups (97.3, 92.3 and 97.4% in C, PRO1 and PRO2 respectively). Memory T-helper lymphocytes increased in V3 vs. V1 (P = 0.032) in PRO2. No differences were found in cytokine concentrations. CONCLUSION: Mixed results have been found regarding the usefulness of Lc supplementation to increase the antigen-specific antibody response to an immune challenge. Clinical trial registration number: EudraCT Number 2016-000183-42. Registered 19 January 2016. Retrospectively registered.

Fatty acid content, health and risk indices, physicochemical composition, and somatic cell counts of milk from organic and conventional farming systems in tropical south-eastern Mexico.

Organic agriculture and livestock farming is claimed to promote animal welfare and can offer animal products with better hygienic-sanitary quality, based on principles of health, ecology, fairness, and care. However, no clear advantages of organic milk (OM) versus conventional milk (CM) from tropical conditions are available. The aims of the study were to determine fatty acid profile, health-promoting (HPI) and thrombogenic (TI) indices, physicochemical composition, and somatic cell counts (SCC) of OM and CM in tropical south-eastern Mexico. Female cross-breed cows (400-600 kg) were employed. CM had larger values of saturated fatty acids (SFA) and polyunsaturated fatty acid (PUFA) (63.6 %; 4.57 %) than OM (61.48 %; 4.22 %), while OM resulted in a larger value of monounsaturated fatty acid (MUFA) (34.3 %) than CM (31.7 %). HPI and TI showed that OM was more favorable than CM. Milk production and physicochemical composition (PC) as well as density had no significant difference, while SCC was significantly lower in OM than in CM on a monthly basis. These results showed that OM promotes a healthful and balanced diet, and is already produced by sustainable ecologic technologies employing traditional agrosilvopastoral management, which is more environmentally friendly and promotes ecological resilience.

Total body water as a possible marker of the altered metabolism in obese children and adolescents / Porcentaje de agua corporal total como posible marcador de alteraciones del metabolismo en niños y adolescentes obesos

Introduction: Obesity is well known to be related to the percentage of Total Body Water (TBW) and inflammation, but the relationship between TBW and inflammation remains still unconfirmed. Aim: To evaluate whether TBW is associated with inflammatory markers in obese children and adolescents. Material and methods: Thirty four obese children (7.0- 10.9 yrs) and 49 obese adolescents (11.0-15.0 yrs) were recruited. Body mass index (BMI Z-score) and several serum biomarkers such as lipid profile, C-Reactive Protein (CRP), sE-selectin, sL-selectin, soluble Intercellular Adhesion Molecule-1 (s-ICAM-1) and soluble Vascular Cell Adhesion Molecule-1 (s-VCAM-1) were determined. TBW was obtained by Bioelectrical measurements. Bilateral partial correlation test was used to analyze associations controlled by sex, age, and/or BMI Z-score. Results: A negative correlation was found between TBW and BMI Z-score in both children and adolescents (r -0.581, p < 0.001; r -0.368, p < 0.011, respectively) that remained in both sex, separately (r -0.540, p < 0.001; r -0.505, p < 0.001). In addition, TBW was also negatively correlated with CRP (r -0.438, p < 0.001) in both groups (r -0.560, p = 0.002; r -0.436, p = 0.007, respectively). When analyzed by sex, TBW was associated with CRP only in boys (r -0.588, p = 0.027; r -0.652, p = 0.005). TBW was negatively correlated with sE-selectin (r -0.236, p = 0.039) only in adolescents (r -0.320, p = 0.032). When analyzed by sex, TBW was associated with sE-selectin only in girls (r -0.432, p = 0.035). Conclusions: TBW may be considered as a marker not only of the hydration status, but also of the metabolic disorder in a low-grade inflammatory process such as obesity in children and adolescents (AU)

Introducción: Se conoce la relación de la obesidad con el porcentaje de agua corporal total (ACT) y con la inflamación, pero no se ha demostrado aún la posible asociación entre ACT y la inflamación. Objetivo: Evaluar si la ACT (como marcador de estado de hidratación) se asocia con marcadores de inflamación en niños y adolescentes obesos. Material y métodos: Los sujetos de este estudio fueron 34 niños obesos (7,0-10,9 años) y 49 adolescentes obesos (11,0-15,0 años). Se determinó el índice de masa corporal (IMC Z-score) y en suero se analizaron perfil lipídico, proteína C-reactiva (PCR), sE-selectina, sL-selectina, las moléculas de adhesión intercelular soluble-1 (s-ICAM-1) y vascular soluble-1 (s-VCAM-1). El ACT se obtuvo mediante impedancia bioeléctrica. Se utilizó la prueba de correlación parcial bilateral para analizar las asociaciones relacionadas con el sexo, la edad y/o IMC Z-score. Resultados: Se observó una correlación negativa entre el ACT y el IMC Z-score en niños y adolescentes (r -0,581, p < 0,001; r -0,368, p < 0,011, respectivamente), que se mantuvo tanto en niños como en niñas (r - 0,540, p < 0,001; r - 0,505, p < 0,001). Además, el ACT también se correlacionó negativamente con la PCR (r -0,438, p < 0,001) en ambos grupos (r -0,560, p = 0,002; r -0,436, p = 0,007, respectivamente). Cuando se analiza por sexo, ACT se asoció con la PCR sólo en los varones (r -0,588, p = 0,027; r -0,652, p = 0,005). ACT se correlacionó negativamente con la sE-selectina (r - 0,236, p = 0,039) solo en el grupo de adolescentes (r -0,320, p = 0.032). Cuando se analiza por sexo, ACT se asoció con la sE-selectina sólo en las niñas (r -0,432, p = 0,035). Conclusiones: ACT puede ser considerado como un marcador no sólo del estado de hidratación, sino también del trastorno metabólico en un proceso inflamatorio de bajo grado como en el caso de la obesidad en niños y adolescentes (AU)

Pharmacotherapeutic Follow-up and Pharmacogenetics of CYP2C9 and CYP3A4 in Antihypertensive Therapy: A Pilot Study in a Community Pharmacy.

No pharmacogenetic studies have yet been conducted in community pharmacies, despite pharmacogenetics being an emerging discipline. Pharmacotherapeutic follow-up (PFU) was performed for 6 months in 37 patients receiving antihypertensive treatment, and they were genotyped for variant alleles *2 and *3 in the CYP2C9 gene and *1B in CYP3A4. Systolic blood pressure, cardiovascular risk, and adherence improved with PFU. Most of the interactions between drugs were represented by concurrent administration of statins and calcium channel blockers, which both use CYP3A4 for their metabolism. Heterozygotic patients for the CYP2C9*2 allele showed higher mean heart rate values after PFU than homozygous patients (73.4 ± 10.0 pulse/min vs 66.2 ± 10.6 pulse/min, respectively; P = .048). Carriers of CYP2C9*2 showed a tendency to less frequent negative outcomes associated with medication due to inefficacy than homozygous patients (50% vs 78.9%, respectively; P = .072). Pharmacogenetics and PFU can be used in community pharmacies to carry out a more exhaustive study of medication in hypertensive outpatients.

Caracterización molecular de dos nuevas mutaciones de α ° talasemia en 2 familias españolas (mutación - - ED y - - GP) / Molecular characterization of two new mutations of α ° thalassemia in two Spanish families (mutation -- ED and -- GP)

Fundamento y objetivo: La hemoglobina es un tetrámero constituido por dos cadenas α y dos cadenas β. Los dos genes estructurales que codifican la cadena α se encuentran localizados en el brazo corto del cromosoma 16. Los individuos normales tienen 4 genes α (αα/αα). Las α talasemias se producen generalmente por la deleción de uno, dos, tres o cuatro de los genes α. La deleción de ambos genes α dentro del mismo cromosoma (α° talasemia) se observa en individuos del área mediterránea y en el sudeste asiático. Material y método: Se estudian dos familias naturales de Madrid con anemia microcítica e hipocroma. El ADN extraído de los leucocitos de sangre periférica se digiere con diferentes enzimas de restricción e hibridación con sondas del cluster de genes α. Los extremos de la deleción se han caracterizado combinando las técnicas de Southern blot, reacción en cadena de la polimerasa (PCR) e hibridación in situ fluorescente (FISH). Resultados: En este trabajo presentamos dos nuevas mutaciones de α° talasemia en dos familias españolas, no descritas previamente en la bibliografía. La mutación (--ED) presenta una deleción de alrededor de 80kb con el punto de rotura 5’ en la coordenada +100 (± 3kb), mientras que el extremo 3’ HVR se sitúa en la coordenada 178±750bp. La segunda mutación (--GP) es más extensa, con pérdida de 145kb, situándose la deleción en la región 5’, entre las coordenadas 34 y 37, respetando el telómero. En la región centromérica la rotura se sitúa también en la coordenada 178±1,4kb. En ambas mutaciones se pierden los dos genes α, el gen θ y la región reguladora HS40. Conclusiones: La exacta identificación de estas mutaciones es esencial para determinar la función de los genes α en caso de consejo genético (AU)

Background and objetives: The two structural genes encoding the human α-globin chains are located on the short arm of chromosome 16. Normal individuals have four genes α (αα/αα). α-thalassemias are usually produced by the deletion of one, two, three, or four α genes. Deletion of both α genes within the same chromosome (α° thalassemia) is commonly observed in individuals from the Mediterranean basin and Southeast Asia. Material and methods: We study two natural families of Madrid with microcytic hypochromic anemia. The DNA extracted from peripheral blood leukocytes was digested with different restriction enzymes and hybridization with probes of gene cluster α. The ends of the deletion were characterized by combining the techniques of Southern blot, PCR and FISH. Results: We present two new mutations of α° thalassemia in two Spanish families, not previously described in the literature. The deletion (--ED) is ∼80kb with the point of bread 5’ in the coordinate +100 (± 3kb), whereas the end 3’HVR places in the coordinate 178±750bp. The second deletion (--GP) is more extensive, with loss of 145kb, placing the deletion in the end 5’ between the coordinates 34 and 37, respecting therefore the telomere. In the centromeric region the point of break places as the previous one in the coordinate 178±1.4bp. Conclusions: In both mutations both alpha genes were deleted, the gene θ and the region HS40. The exact identification of these deletions is essential to determine the function of the genes α with a view to a possible genetic diagnosis (AU)

[Molecular characterization of two new mutations of α° thalassemia in two Spanish families (mutation --(ED) and --(GP))]. / Caracterización molecular de dos nuevas mutaciones de α° talasemia en 2 familias españolas (mutación --(ED) y --(GP)).

BACKGROUND AND OBJECTIVES: The two structural genes encoding the human α-globin chains are located on the short arm of chromosome 16. Normal individuals have four genes α (αα/αα). α-thalassemias are usually produced by the deletion of one, two, three, or four α genes. Deletion of both α genes within the same chromosome (α° thalassemia) is commonly observed in individuals from the Mediterranean basin and Southeast Asia. MATERIAL AND METHODS: We study two natural families of Madrid with microcytic hypochromic anemia. The DNA extracted from peripheral blood leukocytes was digested with different restriction enzymes and hybridization with probes of gene cluster α. The ends of the deletion were characterized by combining the techniques of Southern blot, PCR and FISH. RESULTS: We present two new mutations of α° thalassemia in two Spanish families, not previously described in the literature. The deletion (--(ED)) is ∼80 kb with the break point 5' in the coordinate +100 (± 3 kb), whereas the end 3'HVR places in the coordinate 178±750 bp. The second deletion (--(GP)) is more extensive, with loss of 145 kb, placing the deletion in the end 5' between the coordinates 34 and 37, respecting therefore the telomere. In the centromeric region the breakpoint places as the previous one in the coordinate 178±1.4 bp. CONCLUSIONS: In both mutations both alpha genes were deleted, the gene θ and the region HS40. The exact identification of these deletions is essential to determine the function of the genes α with a view to a possible genetic diagnosis.

Rehabilitation of torture survivors in five countries: common themes and challenges.

BACKGROUND: Torture continues to be a global problem and there is a need for prevention and rehabilitation efforts. There is little available data on torture survivors from studies designed and conducted by health professionals in low income countries. This study is a collaboration between five centres from Gaza, Egypt, Mexico, Honduras and South Africa who provide health, social and legal services to torture survivors, advocate for the prevention of torture and are part of the network of the International Rehabilitation Council for Torture Victims (IRCT). METHODS: Socio-demographic, clinical and torture exposure data was collected on the torture survivors attending the five centres at presentation and then at three and six month follow-up periods. This sample of torture survivors is presented using a range of descriptive statistics. Change over time is demonstrated with repeated measures analysis of variance. RESULTS: Of the 306 torture survivors, 23% were asylum seekers or refugees, 24% were socially isolated, 11% in prison. A high level of traumatic events was experienced. 64% had suffered head injury whilst tortured and 24% had ongoing torture injury problems. There was high prevalence of symptoms of anxiety, depression, post traumatic stress as well as medically unexplained somatic symptoms. The analysis demonstrates a modest drop in symptoms over the six months of the study. CONCLUSIONS: Data showed that the torture survivors seen in these five centres had high levels of exposure to torture events and high rates of clinical symptoms. In order to provide effective services to torture survivors, health professionals at torture rehabilitation centres in low income countries need to be supported to collect relevant data to document the needs of torture survivors and to evaluate the centres' interventions.

Human promoter mutations unveil Oct-1 and GATA-1 opposite action on Gfi1b regulation.

Growth factor-independence 1b (Gfi1b) is a zinc finger transcription factor essential for erythroid and megakaryocytic development. To better understand Gfi1b regulation and to know the implication of the level of expression of this gene in human pathology, we have searched for promoter punctual sequence variations in 214 patients with different hematological diseases. We found two previously unknown congenital mutations at evolutionary conserved GATA and octamer-binding (Oct) transcription factor sites. The Oct site mutation was also found in five relatives of the patient. The GATA motif mutation reduced promoter activity by 50% in vitro, while homozygous patients with the octamer site mutation showed a four-to-five times increase of Gfi1b RNA in platelets. Electrophoretic mobility shift analyses demonstrated that different protein complexes bind to both sites and that binding is reduced by the mutations. Finally, we found that GATA-1 and Oct-1 are the main components of each complex. This study provides evidences of a new mechanism for Gfi1b repression. This is also the first report of Gfi1b mutations with a functional implication; further investigation and follow-up will clarify the involvement of these mutations in hematological disease.