miR-328 mediates a metabolic shift in colon cancer cells by targeting SLC2A1/GLUT1

Purpose: Increasing evidence shows that altered metabolism is a critical hallmark in colon cancer. There is a strong need to explore the molecular mechanisms underlying cancer metabolism. Whether the aberrant expression of microRNAs contributes to cancer metabolism is not fully understood. miR-328 is a putative potential target of SLC2A1, but the regulating mechanism between them remains unknown. We have examined whether miR-328 directly regulates SLC2A1/GLUT1 expression in colon cancer cells. Methods: We performed in silico bioinformatic analyses to identify miR-328-mediated molecular pathways and targets. We also performed luciferase assays and western blot analyses in LOVO and SW480 colon cancer cell lines. In addition, we assessed miR-328 expression in 47 paired tumor and normal tissue specimens from resected colon cancer patients. Results: Luciferase reporter assays showed that miR-328 directly targeted SLC2A1 3′-untranslated region (UTR), with a significant decrease in luciferase activity in both LOVO and SW480 cell lines. These results were validated by western blot. miR-328 expression was significantly downregulated in tumor tissue compared with paired normal tissue. Conclusions: Our results show that miR-328 targets SLC2A1/GLUT1. We suggest that miR-328 may be involved in the orchestration of the Warburg effect in colon cancer cells. Furthermore, miR‐328 expression is reduced in colon cancer patients and thus inversely correlates with the classically reported upregulated SLC2A1/GLUT1 expression in tumors

No disponible

Tratamiento farmacológico del asma / Pharmacological treatment of asthma

El asma se considera un síndrome que agrupa diferentes formas de enfermedad, en la que factores genéticos y ambientales interaccionan y generan manifestaciones de la enfermedad. Su expresión clínica es muy variable: desde síntomas agudos y esporádicos a crónicos, desde estacionales o que aparecen en relación con el ejercicio, hasta una enfermedad grave y persistente. En España afecta a uno de cada diez niños, con amplias variaciones regionales. En tratamiento del asma nos encontramos con dos situaciones clínicas: 1) las sibilancias recurrentes en lactantes y preescolares son un motivo frecuente de consulta y el retraso en el inicio del tratamiento en muchos casos puede tener gran impacto sobre la salud del paciente a largo plazo. Algunos niños serán asmáticos que inician los síntomas precozmente, pero otros tendrán sibilancias que desaparecerán en la edad escolar. Por tanto, no siempre es fácil diagnosticarla y decidir el tratamiento en estos casos. 2) La elección del tratamiento farmacológico en el niño mayor de cinco años debe de hacerse en base a la evidencia clínica disponible y en ese sentido las propuestas de la GINA y de la guía de British Thoracic Society, Scottish Intercollegiate Guidelines Network (SIGN), modificadas siguiendo recomendaciones de la guía de práctica clínica sobre asma infantil. El conocimiento y la utilización de estas guías y consensos con las recomendaciones actuales para el tratamiento de mantenimiento y de la crisis de asma en la infancia, son junto con la educación basada en el autocontrol, puntos básicos de un programa de atención al niño y adolescente con asma

Asthma is considered a syndrome that groups different forms of disease, in which genetic and environmental factors interact and generate manifestations of the disease. Its clinical expression is very variable: from acute and sporadic symptoms to chronic, from seasonal or appearing in relation to exercise, to a serious and persistent disease. In Spain it affects one in 10 children with wide regional variations. In asthma treatment we find two clinical situations: 1) recurrent wheezing in infants and pre-schoolers is a frequent reason for consultation and delay in the onset of treatment in many cases can have a great impact on the health of the patient in the long run. Some children will be asthmatics who start symptoms early, but others will have wheezing that disappears in school age. Therefore, it is not always easy to diagnose and decide the treatment in these cases. 2) The choice of pharmacological treatment in the child over five years old should be based on the clinical evidence available and in this sense the proposals of the GINA and the guide British Thoracic Society, Scottish Intercollegiate Guidelines Network (SIGN) modified following recommendations of the child asthma CPG. The knowledge and use of these guidelines and consensus with current recommendations for the treatment of maintenance and the asthma crisis in childhood, are along with education based in self-control, basic points of a program of care for children and adolescents with asthma

miR-328 mediates a metabolic shift in colon cancer cells by targeting SLC2A1/GLUT1.

PURPOSE: Increasing evidence shows that altered metabolism is a critical hallmark in colon cancer. There is a strong need to explore the molecular mechanisms underlying cancer metabolism. Whether the aberrant expression of microRNAs contributes to cancer metabolism is not fully understood. miR-328 is a putative potential target of SLC2A1, but the regulating mechanism between them remains unknown. We have examined whether miR-328 directly regulates SLC2A1/GLUT1 expression in colon cancer cells. METHODS: We performed in silico bioinformatic analyses to identify miR-328-mediated molecular pathways and targets. We also performed luciferase assays and western blot analyses in LOVO and SW480 colon cancer cell lines. In addition, we assessed miR-328 expression in 47 paired tumor and normal tissue specimens from resected colon cancer patients. RESULTS: Luciferase reporter assays showed that miR-328 directly targeted SLC2A1 3'-untranslated region (UTR), with a significant decrease in luciferase activity in both LOVO and SW480 cell lines. These results were validated by western blot. miR-328 expression was significantly downregulated in tumor tissue compared with paired normal tissue. CONCLUSIONS: Our results show that miR-328 targets SLC2A1/GLUT1. We suggest that miR-328 may be involved in the orchestration of the Warburg effect in colon cancer cells. Furthermore, miR-328 expression is reduced in colon cancer patients and thus inversely correlates with the classically reported upregulated SLC2A1/GLUT1 expression in tumors.

Crisis de asma / Asthma crisis

No disponible

Educación en el niño y adolescente con asma (2ª parte) / Education in children and adolescents with asthma (part 2)

No disponible

miR-203 and miR-221 regulate SOCS1 and SOCS3 in essential thrombocythemia.

The biological basis of essential thrombocythemia (ET) patients lacking known mutations is still unknown. MicroRNAs (miRNA) regulate hematopoietic differentiation and are deregulated in several hematopoietic malignancies. However, miRNA expression in ET patients has been poorly explored. We performed miRNA profiling in platelets from 19 ET patients and 10 healthy controls. Hierarchical cluster analysis showed two well-separated clusters between patients and controls, indicating that ET platelets had a characteristic 70-miRNA signature (P<0.0001), 68 of which were downregulated. According to the mutational status, three differentially expressed miRNAs, miR-15a (P=0.045), miR-150 (P=0.001) and miR-519a (P=0.036), were identified. A 40-miRNA signature was identified characterizing JAK2V617F-positive ET patients. Eight genes, whose interaction with the miRNAs could activate the JAK/STAT pathway were identified. An inverse correlation was observed between miRNAs expression and their target genes for SOCS1 and miR-221, SOCS3 and miR-221, SOCS3 and miR-203, and PTPN11 and miR-23a. All three miRNAs were upregulated in JAK2V617F-negative ET patients. SOCS1 and SOCS3 were validated as targets of miR-221 and miR-203, respectively. In summary, our study shows that platelets from JAK2V617F-negative ET patients harbor a specific miRNA signature that can participate in the modulation of the JAK/STAT pathway through regulation of key genes as SOCS1 and SOCS3.

Quality of life in caregivers of children with asthma: Validity and reliability of the IFABI-R questionnaire.

BACKGROUND: Parents'/caregivers' quality of life is an important aspect to consider when handling paediatric asthma, but there is a paucity of valid and reliable instruments to measure it. The Family Impact of Childhood Bronchial Asthma (IFABI-R) is a recently developed questionnaire to facilitate the assessment of asthma-related parents'/caregivers' quality of life. This study researches the psychometric properties of IFABI-R. METHODS: Parents/main caregivers of 462 children between 4 and 14 years of age with active asthma were included in the sample. IFABI-R was administered on two different occasions and a number of other variables related to the parents'/caregivers' quality of life were measured: child's asthma control, family functioning, and parents'/caregivers' perception of asthma symptoms in the child. IFABI-R evaluative and discriminative properties were analysed, and the minimal important change in the IFABI-R score was identified. RESULTS: IFABI-R showed high internal consistency (Cronbach's alpha=0.941), cross-sectional construct validity (correlation with the degree of child's asthma control, family functioning and parent/caregiver perception of the child's asthma symptoms), longitudinal construct validity (correlation of changes in the IFABI-R with changes in asthma control and changes in the perception of symptoms), sensitivity to change and test-retest reliability. An absolute change of 0.3 units in IFABI-R related to a minimal significant change in the parents'/caregivers' quality of life. CONCLUSIONS: IFABI-R is a reliable and valid instrument to study the quality of life of parents/caregivers of children with asthma.

The expression level of BAALC-associated microRNA miR-3151 is an independent prognostic factor in younger patients with cytogenetic intermediate-risk acute myeloid leukemia.

Acute myeloid leukemia (AML) is a heterogeneous disease whose prognosis is mainly related to the biological risk conferred by cytogenetics and molecular profiling. In elderly patients (⩾60 years) with normal karyotype AML miR-3151 have been identified as a prognostic factor. However, miR-3151 prognostic value has not been examined in younger AML patients. In the present work, we have studied miR-3151 alone and in combination with BAALC, its host gene, in a cohort of 181 younger intermediate-risk AML (IR-AML) patients. Patients with higher expression of miR-3151 had shorter overall survival (P=0.0025), shorter leukemia-free survival (P=0.026) and higher cumulative incidence of relapse (P=0.082). Moreover, in the multivariate analysis miR-3151 emerged as independent prognostic marker in both the overall series and within the unfavorable molecular prognostic category. Interestingly, the combined determination of both miR-3151 and BAALC improved this prognostic stratification, with patients with low levels of both parameters showing a better outcome compared with those patients harboring increased levels of one or both markers (P=0.003). In addition, we studied the microRNA expression profile associated with miR-3151 identifying a six-microRNA signature. In conclusion, the analysis of miR-3151 and BAALC expression may well contribute to an improved prognostic stratification of younger patients with IR-AML.

Atención en las primeras 48 horas tras el alta de maternidad en la consulta de enfermería o Pediatría / Assistance on the first 48 hours after discharge from maternity in Primary Care Practice

Introducción: desde el momento en que el recién nacido es dado de alta de la maternidad hasta que es atendido en su centro de salud (CS) pasa un periodo de tiempo crucial y, a veces, excesivamente prolongado. Objetivo: conocer la edad en que son atendidos por primera vez los recién nacidos en Atención Primaria (AP) en la Comunidad Valenciana. Material y métodos: encuesta dirigida a los pediatras de AP. Resultados: participaron 46 CS de las provincias de Castellón y Valencia (de un total de 162 centros). Se declararon datos de 248 recién nacidos durante el periodo del 9 de abril al 30 de junio del año 2013. El 58% de los recién nacidos fueron vistos por primera vez en su CS a partir del quinto día de vida. Los niños nacidos en un hospital público tienen casi dos veces más probabilidades de acudir al CS antes de los seis días de vida que los niños nacidos en un hospital privado (odds ratio [OR]: 1,97; intervalo de confianza del 95% [IC 95%]: 0,92 a 4,1; p=0,07). El 56,93% de las lactancias maternas y mixtas fueron vistas después del quinto día de vida. El 24,2% de los casos estudiados no tenía la tarjeta de asistencia sanitaria (TAS) al salir de la maternidad. La mayoría de los niños nacidos en maternidades privadas no tenía la TAS al alta, requisito imprescindible para solicitar cita en su CS. Conclusiones: desde el momento en que el recién nacido es dado de alta de la maternidad hasta que es atendido por primera vez por el equipo de pediatra-enfermera, pasa un periodo de tiempo importante (AU)

Introduction: a critical period of time, sometimes too long, passes since healthy term infants are discharged from hospital until they are visited at primary care practices. Objective: to assess the age after birth when healthy term infants are first checked at primary care practices in Comunitat Valenciana. Material and methods: survey of primary care pediatricians from Comunitat Valenciana. Results: Forty-six Primary Care Practices participated in the provinces of Castellón and Valencia (from a total of 162 practices). Data from 248 term infants from April 9th to June 30th 2013 were collected. Fifty-eight percent of the infants were first seen after the fifth day of life. Children born in a public hospital are almost twice as much likely to be visited before 6 days of life than children born in a private hospital (odds ratio [OR]: 1.97, 95% CI: 0.92 to 4.1, p = 0.07). Exclusive breastfeeding and mixed feeding were seen after the fifth day of life in 56.93%. No health insurance card at discharge (most of children born in a private maternity), essential to make an appointment, was observed in 24.2% of the cases studied. Conclusions: since the time healthy term infants are discharged from hospital until they are first visited by the pediatric-nurse team, a too long critical period passes

MicroRNA expression at diagnosis adds relevant prognostic information to molecular categorization in patients with intermediate-risk cytogenetic acute myeloid leukemia.

Acute myeloid leukemia (AML) is a heterogeneous disease, and optimal treatment varies according to cytogenetic risk factors and molecular markers. Several studies have demonstrated the prognostic importance of microRNAs (miRNAs) in AML. Here we report a potential association between miRNA expression and clinical outcome in 238 intermediate-risk cytogenetic AML (IR-AML) patients from 16 institutions in the CETLAM cooperative group. We first profiled 670 miRNAs in a subset of 85 IR-AML patients from a single institution and identified 10 outcome-related miRNAs. We then validated these 10 miRNAs by individual assays in the total cohort and confirmed the prognostic impact of 4 miRNAs. High levels of miR-196b and miR-644 were independently associated with shorter overall survival, and low levels of miR-135a and miR-409-3p with a higher risk of relapse. Interestingly, miR-135a and miR-409-3p maintained their independent prognostic value within the unfavorable molecular subcategory (wild-type NPM1 and CEBPA and/or FLT3-ITD), and miR-644 retained its value within the favorable molecular subcategory. miR-409-3p, miR-135a, miR-196b and mir-644 arose as prognostic markers for IR-AML, both overall and within specific molecular subgroups.

Acute myeloid leukemia with translocation (8;16)(p11;p13) and MYST3-CREBBP rearrangement harbors a distinctive microRNA signature targeting RET proto-oncogene.

Acute myeloid leukemia (AML) with t(8;16)(p11;p13) (t(8;16) AML) has unique clinico-biological characteristics, but its microRNA pattern is unknown. We analyzed 670 microRNAs in seven patients with t(8;16) AML and 113 with other AML subtypes. Hierarchical cluster analysis showed that all t(8;16) AML patients grouped in an independent cluster. Supervised analysis revealed a distinctive signature of 94-microRNAs, most of which were downregulated, including miR-21 and cluster miR-17-92. The mRNA expression analysis of two known transcription factors of these microRNAs (STAT3 and c-Myc, respectively) showed significant downregulation of STAT3 (P=0.04). A bioinformatic analysis showed that 29 of the downregulated microRNAs might be regulated by methylation; we treated a t(8;16) AML sample with 5-aza-2'-deoxycytidine (5-AZA-dC) and trichostatin A and found that 27 microRNAs were re-expressed after treatment. However, there was no difference in methylation status between t(8;16) and other AML subtypes, either overall or in the microRNA promoter. Cross-correlation of mRNA and microRNA expression identified RET as a potential target of several microRNAs. A Renilla-luciferase assay and flow cytometry after transfection with pre-microRNAs confirmed that RET is regulated by miR-218, miR-128, miR-27b, miR-15a and miR-195. In conclusion, t(8;16) AML harbors a specific microRNA signature that is partially epigenetically regulated and targets RET proto-oncogene.

Expression of myosin heavy chain isoforms in the human supraspinatus muscle: variations related to age and sex.

The contractile function of skeletal muscles is primarily regulated by the expression of myosin heavy chain (MHC) isoforms. Adult human skeletal muscles express three MHC isoforms (MHC-I, MHC-IIa and MHC-IIx). The muscles mainly expressing MHC-I are slow but resistant to fatigue, while those with major expression of MHC-IIa and MHC-IIx are fast and powerful but less resistant to fatigue. In this study, mRNA levels of the MHC isoforms were assessed in 24 human supraspinatus muscles by reverse-transcription polymerase chain reaction. The average expression of the MHC-I isoform was 36.72%, that of the MHC-IIa isoform was 33.52%, and the average expression of the MHC-IIx isoform was 29.76%. The higher average expression of the two MHC-II isoforms taken together (63.28%) indicates that the human supraspinatus muscle is a powerful, fast muscle with relatively low resistance to fatigue, in accordance with its role in the elevation of the upper extremity. In women, and more markedly in older women, the trend towards upregulation of the fast MHC-II isoforms and downregulation of the slow MHC-I isoform, which is absent in males, may improve our understanding of possible causes of the subacromial impingement syndrome.

Claves de educación en asma: casos clínicos interactivos / Keys of education in asthma: interactive cases report

El asma es una enfermedad crónica compleja, con una gran variabilidad y que tiene un enorme impacto, no solo en los pacientes afectados y sus familias, sino también en la sociedad en general. La prevalencia de asma en la infancia es elevada, afecta alrededor del 10% de la población infantil y adolescente, siendo en nuestro medio la patología crónica que consume un mayor número de recursos y tiempo en la Atención Primaria pediátrica. El manejo adecuado del asma implica que se realice un diagnóstico correcto, se evalúe la gravedad y se prescriba el tratamiento indicado. Un componente esencial en el tratamiento del asma infantil es la educación terapéutica, en la que el paciente y su familia han de ser formados y educados, con la finalidad de conseguir un buen control de la enfermedad y mejorar la calidad de vida. Para que los programas educativos sean efectivos, es necesaria una adecuada formación de los profesionales de la salud, siendo importante una buena comunicación y relación entre sanitarios y pacientes para conseguir que estos entiendan la enfermedad y su tratamiento, y mejoren la adherencia. En este seminario se van a presentar una serie de casos clínicos para desarrollar diversos contenidos diagnósticos, terapéuticos y educativos del asma infantil (AU)

Asthma is a chronic complex disease, with a great variability and a big impact, not only in the affected patients and their families, but also in general society. The prevalence of childhood asthma is high, it affects about 10% of the children and adolescents, and it is the chronic condition that consumes more resources and time in pediatric primary care in our population. The correct asthma management implies that a proper diagnosis is made, that the seriousness of the condition has been evaluated and that the appropriate treatment has been prescribed. Therapeutic education is an essential component in the treatment of childhood asthma in which the patient and his family have to be trained and educated, in order to achieve a good control of the disease and improve the quality of life. In order that educational programs are effective it is necessary the adequate training of health professionals, being important an adequate communication and relationship between them and the patients so that they understand the disease, its treatment and they improve the adherence to treatment. This seminar will present a series of clinical cases to approach various diagnostic, therapeutic and educational contents of childhood asthma (AU)

Surgical technique in cadaveric donors for partial hand allotransplant in patients with rheumatoid arthritis.

For patients with severe hand deformities due to rheumatoid arthritis, we propose an allotransplantation of an osteomyotendinose structure (OMTS), preserving the recipient's skin and sensory nerves. Our objective was to develop the surgical technique in a 10 cadavers, five as donors and five as recipients. The donor's hand was 10% to 15% smaller than the recipient's. Dissections were performed by two surgical teams under magnification. In the donor, the OMTS was procured at the distal third of the forearm, maintaining the integrity of the arterial system, with its concomitant veins and motor branches of the median and ulnar nerves, leaving the skin envelope. In the recipient, the OMTS was removed, taking care to preserve the cutaneous cover with the digital arteries in continuity with the superficial palmar arch and radial and ulnar arteries. Also, the digital nerves were maintained in the skin flap, in continuity with the median and ulnar nerves. Their motor branches were divided after emergence from the main nerves. The superficial dorsal veins and radial nerve were kept adhered to the cutaneous cover. Then, the donor OMTS was placed within the recipient cutaneous flap; all the anatomic structures were repaired. The average surgical time was 780 minutes. Methylene blue was present in the digital arteries. There were no difficulties in the anatomic repair. The surgical technique is quite laborious, especially the dissection of the recipient interdigital spaces. Due to the requirement for arterial system integrity, the cutaneous flap must be viable. Also, the allotransplanted OMTS has all necessary conditions to obtain good tissue perfusion for subsequent function. Procurement without skin permits a greater opportunity to find donors, and greater social and personal acceptance by the recipient.

Tumour CD133 mRNA expression and clinical outcome in surgically resected colorectal cancer patients.

BACKGROUND: Human prominin-1 (CD133) is a novel pentaspan membrane protein which was originally classified as a marker of primitive haematopoietic and neural stem cells. Cancer stem cells have been isolated and expanded from leukaemia and several solid tumours, and have been associated with metastasis, chemoresistance and relapse. CD133 is recognised as a stem cell marker and is capable of identifying a tumour-initiating subpopulation in brain, colon, melanoma and other solid tumours. METHODS: We assessed CD133 mRNA expression levels by RT-QPCR in tumour and matched normal tissue from 64 stages I-III colorectal cancer (CRC) patients and correlated tumour CD133 levels with clinicopathological characteristics and clinical outcome. RESULTS: In four patients, CD133 mRNA was not expressed in tumour or in normal tissue. In the remaining 60 patients, expression levels were higher in tumour than in normal tissue (p=0.001). Higher levels of CD133 expression were associated with shorter relapse-free interval (RFI) (p=0.004) and overall survival (OS) (p<0.0001). In the multivariate analyses, CD133 levels emerged as a prognostic marker for RFI and OS. CONCLUSIONS: We have observed longer RFI and OS in patients with lower levels of CD133, regardless of adjuvant treatment and other clinical characteristics. If these findings are confirmed in larger prospective studies, CD133 assessment may prove useful for new diagnostic and therapeutic procedures for CRC patients.

Epigenetic events in normal colonic mucosa surrounding colorectal cancer lesions.

Gene inactivation by promoter hypermethylation has been demonstrated in the colonic mucosa of colorectal cancer (CRC) patients. However, current data do not prove direct involvement of this epigenetic modification in the early stages of CRC. Promoter methylation profiles of E-cadherin, hMLH1, MGMT, p16(INK4a), p15(INK4b) and p14(ARF); mutations of K-ras, B-raf and TP53 and microsatellite instability (MSI) were examined in normal and cancerous colonic mucosal tissue in 82 CRC patients using methylation-specific PCR assays. Methylation of hMLH1 and MGMT in normal mucosa correlated significantly with MSI and K-ras activation in neighbouring cancerous mucosal tissues. Similarly, poorly differentiated tumours were associated with methylated p16(INK4a) and E-cadherin in neighbouring normal colonic tissues (NCTs). Our results indicate that epigenetic changes in mucosa surrounding colorectal neoplastic lesions may describe a 'field cancerisation' phenomenon that may occur previous to genetic alterations in early stages of carcinogenesis.

Pharmacogenomics: a tool for improving cancer chemotherapy

When an anticancer drug is captured and transported to the interior of a cell, many biochemical processes intervene in the metabolism of the drug. First, the Phase I and Phase II enzymes produce metabolites, which are then excreted via the ABC-transporter enzymes. Finally, the remaining drug binds to its molecular target. Genetic alterations known as polymorphisms in any of the proteins that intervene in these processes can affect the efficacy of treatment. Interestingly, these polymorphisms can be detected in both normal and tumour cells. In this article, we will review the most effective method of detecting single-nucleotide polymorphisms (SNPs) and describe the principal SNPs in enzymes involved in drug metabolism and in DNA repair. Finally, we will briefly describe promising lines of future research in this field (AU)

No disponible

Identification by Real-time PCR of 13 mature microRNAs differentially expressed in colorectal cancer and non-tumoral tissues.

MicroRNAs (miRNAs) are short non-coding RNA molecules playing regulatory roles by repressing translation or cleaving RNA transcripts. Although the number of verified human miRNA is still expanding, only few have been functionally described. However, emerging evidences suggest the potential involvement of altered regulation of miRNA in pathogenesis of cancers and these genes are thought to function as both tumours suppressor and oncogenes. In our study, we examined by Real-Time PCR the expression of 156 mature miRNA in colorectal cancer. The analysis by several bioinformatics algorithms of colorectal tumours and adjacent non-neoplastic tissues from patients and colorectal cancer cell lines allowed identifying a group of 13 miRNA whose expression is significantly altered in this tumor. The most significantly deregulated miRNA being miR-31, miR-96, miR-133b, miR-135b, miR-145, and miR-183. In addition, the expression level of miR-31 was correlated with the stage of CRC tumor. Our results suggest that miRNA expression profile could have relevance to the biological and clinical behavior of colorectal neoplasia.

Trastornos genéticos asociados a las alteraciones del número de los dientes. Estado de la cuestión / No disponible

El presente estudio representa una revisión de la litaratura (..) (AU)

The present study represents a revision of the (..) (AU)