Biomarker profile predicts clinical efficacy of extracorporeal photopheresis in steroid-resistant acute and chronic graft-vs-host disease after allogenic hematopoietic stem cell transplant.

We conducted a multicenter interventional study to assess the efficacy of Therakos ECP to treat steroid-resistant graft-vs-host disease (SRes-GVHD) after allogeneic HSCT and to identify biomarkers of GVHD response. A total of 62 patients were treated for acute SRes-GVHD (n = 37) or chronic SRes-GVHD (n = 25). Median time to best response was 35 days (range, 28-85) and 90 days (range, 27-240) in acute and chronic SRes-GVHD, respectively. Overall, 27 patients (72.9%) with SRes-aGVHD responded to treatment (40.5% CR and 32.4% PR). The response rate was significantly higher in grade I-II than in grade III-IV aGVHD (100% vs 50.0%, respectively, P-value = .001). In chronic SRes-GVHD, 22 patients (88%) achieved a clinical response (24.0% CR and 64% PR). Response was higher in moderate than in severe SRes-cGVHD (100% vs 75%, P = .096). In both acute and chronic SRes-GVHD patients, the percentage of peripheral blood CD3+ CD4+ was higher and CD3+ CD8+ lower in responding than nonresponding patients. Acute SRes-GVHD responding patients presented a higher number of Treg cells (CD4+ CD25+ CD127low/- ) at day 0 (P = .028) than nonresponding patients, differences that were maintained over the observation period. Phenotypic analysis of T-cell maturation showed a trend toward reduction in TCD8 naive cells, along with an increased percentage of TCD8 Mem Efect T cells after starting ECP in responding patients. None of the studied serum cytokines displayed statistically significant changes in either acute or chronic SRes-GVHD. ECP is an effective treatment for patients with SRes-GVHD. Biomarkers could help guide decision-making on ECP treatment initiation and duration.

Clustering of lifestyle factors in Spanish university students: the relationship between smoking, alcohol consumption, physical activity and diet quality.

OBJECTIVE: To ascertain the prevalence of and association between main lifestyle factors (diet, physical activity, alcohol consumption and smoking) in students from the Balearic Islands University. DESIGN: A cross-sectional, descriptive study. A questionnaire including questions on lifestyle, dietary habits and physical activity habits was administered to the students. Four different diet quality scores were calculated (Diet Diversity Score, Mediterranean Diet Score, Dietary Guidelines Score and Global Dietary Guidelines Score). SETTING: A sample of students from the Balearic Islands University. SUBJECTS: Nine hundred and eighty-seven students (45·5 % males; mean age 21·5 (sd 3·3) years). RESULTS: The dietary pattern of the student population was characterized by a low consumption of cereals and tubers, fruits, vegetables, olive oil, legumes and nuts, and a high consumption of processed meat, sweets, snacks, soft drinks and pastries. Linear, positive and statistically significant correlations were found between the number of meals consumed daily and all of the diet quality scores determined. Determinants of diet quality, both in the univariate and multivariate analyses, were physical activity practice, sex, age and number of meals consumed daily. CONCLUSIONS: Risk factors such as smoking, diet and physical inactivity had a tendency of clustering among Spanish university students. Overall diet quality was low, due to important departures from dietary recommendations and loss of the traditional Mediterranean dietary pattern. Nutritional education campaigns that include promotion of physical activity practice are needed to improve the overall health status of this population.

Early metal pollution in southwestern Europe: the former littoral lagoon of El Almarjal (Cartagena mining district, S.E. Spain).A sedimentary archive more than 8000 years old.

A borehole drilling campaign has allowed the study of a former littoral lagoon located next to the harbour city of Cartagena in South-East Spain (close to the Sierra de Cartagena polymetallic ore deposits). This lagoon, which developed during the Holocene, was first a shallow sedimentary marine environment and later evolved into a swampy semi-endorheic basin named "Almarjal" (after the Arab term from the fourteenth century). The lagoon eventually dried out and at present forms part of the substratum of the modern sector of the city urban area. The basin representative sediments are sapropelic black silty facies forming a continuous sedimentary archive, accounting for more than 8000 years of depositional phenomena. The geochemical study of these sediments, together with their absolute calibrated dating by 14C, allows definition of successive stages of mining and metallurgical activities in the area. In turn, this information provides a more comprehensive perspective regarding metal pollution, particularly lead contamination during different periods of the Recent Prehistory and the Classical Age. The results indicate that the beginning of contamination by lead and other heavy metals occurred as early as 4500 years ago, when the Final Chalcolithic period was taking place in the South-East of the Iberian Peninsula. This finding provides further insights regarding the debate on the origins of lead mining and metallurgy in SE Spain.

Long-term cultured human myotubes decrease contractile gene expression and regulate apoptosis-related genes.

The present study examined time-dependent changes in the gene expression profile of long-term cultured human myotubes. Microarray transcriptional analysis was performed in a primary culture of differentiated myotubes from one subject over seven weeks. This analysis showed a main gradual fall in genes of the contractile apparatus, and a broad upregulation of genes involved in cell development and growth, followed by stress response and signal transduction. Glucose metabolism was also monitored, but no significant alterations in glucose uptake, oxidation or glycogen storage were observed. Mitochondrial membrane potential, or the amount of membrane lipid peroxides, remained similarly unchanged, nor was lactate dehydrogenase leakage observed. Time-dependent changes in eight genes were validated by real-time RT-PCR in primary cultured myotubes from four subjects, of similar age and isolated after equivalent replication cycles in vitro and differentiated over seven weeks. Insulin-like growth factor-binding protein 2 (IGFBP2), a modulator of the IGF signal, was upregulated. The antiapoptotic gene heat-shock 70-kd protein 2 (HSPA2) was induced, whereas the proapoptotic tumor necrosis factor receptor superfamily, member 25 (WSL-1) was suppressed. A decline in the muscle-specific gene M-cadherin and contraction genes, such as slow-twitch troponin I (TNNI1) and myosin heavy chain 2 (MYH2), myosin light chain 1 (MYL1) and myosin-binding protein H (MYBPH), which are expressed in adult fast-twitch muscle, was shown. In summary, these data demonstrate extensive downregulation of contractile genes and modulation of apoptosis-related genes, in favour of cell survival, during maintenance of cultured human myotubes.

Effect of sucrose and saturated-fat diets on mRNA levels of genes limiting muscle fatty acid and glucose supply in rats.

In this study, we examined whether the increased availability of lipids in blood resulting from two types of diet manipulation regulated metabolic gene expression in the skeletal muscle of rats. Feeding for 4 wk on an isocaloric-sucrose or a hypercaloric-fat diet increased plasma TAG in the fed condition by increments of 70 and 40%, respectively, and increased fasting insulinemia (approximately 3-fold) compared with a starch diet. The fat diet impaired glucose tolerance and caused obesity, whereas sucrose-fed rats maintained their normal weight. We analyzed the expression of genes that regulate the exogenous FA supply (LPL, FAT/CD36, FATP1), synthesis (ACC1), glucose (GLUT4, GLUT1, HK2, GFAT1, glycogen phosphorylase) or glycerol (glycerol kinase) provision, or substrate choice for oxidation (PDK4) in gastrocnemius and soleus muscles at the end of the glucose tolerance test. LPL, FAT/CD36, FATP1, PDK4, and GLUT4 mRNA as well as glycogen phosphorylase and glycerol kinase activity levels in both muscles were unchanged by the diets. Increased mRNA levels of GLUT1 (1.6- and 2.6-fold, respectively) and GFAT1 (about 1.7-fold) in gastrocnemius, and of ACC1 (about 1.5-fold) in soleus, were found in both the sucrose and fat groups. In the fat group, HK2 mRNA was also higher (1.8-fold) in the gastrocnemius. Both sucrose and saturated-fat diets prompted hyperinsulinemia and hyperlipemia in rats. These metabolic disturbances did not alter the expression of LPL, FAT/CD36, FATP1, PDK4, and GLUT4 genes or glycogen phosphorylase and glycerol kinase activity levels in either analyzed muscle. Instead, they were linked to the coordinated upregulation in gastrocnemius of genes that govern glucose uptake and the hexosamine pathway, namely, GLUT1 and GFAT1, which might contribute to insulin resistance.

[Prevalence of multiple sclerosis in the Region of Murcia]. / Prevalencia de la esclerosis multiple en la Region de Murcia.

INTRODUCTION: Multiple sclerosis (MS) is a demyelinating inflammatory disease of the central nervous system with immune-mediated pathogenesis. Recent research points to an increase in its prevalence, and a number of studies relate Epstein-Barr virus (EBV) with its aetiology. AIMS: This study seeks to analyse the prevalence of MS in the Region of Murcia, and includes a description of the clinical characteristics at the time of onset of the disease, and of the EBV serological status of patients with MS. PATIENTS AND METHODS: We conducted a retrospective epidemiological study based on a sample consisting of the population living within the central-west healthcare area of the Region of Murcia (257,865 inhabitants). Clinical and serological data extracted from different sources were analysed. RESULTS: Prevalence of MS in the population under study: 88 cases/100,000 inhabitants. Prevalence of MS together with isolated demyelinating syndrome: 98.4 cases/100,000 inhabitants. Mean incidence of MS: 5.8 cases/100,000 inhabitants/year. At the onset of MS, 67.8% were females, 81.9% presented a relapsing-remitting course, the mean age was 31.4 years, the sensory system was the most frequently compromised (45.1%), onset was monofocal in 55.4% and the degree of disability on the Expanded Disability Status Scale was 2.1 points. The seroprevalence of EBV was 99.3%. The reactivation of EBV infection was related to the clinical activity of MS in 10 patients (45.4%). CONCLUSIONS: Currently, the prevalence of MS in the Region of Murcia is similar to that estimated in other Spanish autonomous regions. The study confirms the trend of increased prevalence observed over the last few decades.

TITLE: Prevalencia de la esclerosis multiple en la Region de Murcia.Introduccion. La esclerosis multiple (EM) es una enfermedad inflamatoria desmielinizante del sistema nervioso central con patogenia inmunomediada. Recientes estudios indican un aumento de su prevalencia, y numerosos trabajos relacionan el virus de Epstein-Barr (VEB) con su etiologia. Objetivo. Analisis de prevalencia de la EM en la Region de Murcia, incluyendo la descripcion de las caracteristicas clinicas en el momento del inicio de la enfermedad, y del estado serologico del VEB de los pacientes con EM. Pacientes y metodos. Estudio epidemiologico retrospectivo, tomando como muestra la poblacion residente en el area sanitaria centro-oeste de la Region de Murcia (257.865 habitantes). Se analizan datos clinicos y serologicos extraidos de diferentes fuentes. Resultados. Prevalencia de la EM en la poblacion estudiada: 88 casos/100.000 habitantes. Prevalencia de la EM junto con el sindrome desmielinizante aislado: 98,4 casos/100.000 habitantes. Incidencia media de la EM: 5,8 casos/100.000 habitantes/año. En el inicio de la EM, el 67,8% eran mujeres, el 81,9% presentaba un curso recurrente-remitente, la edad media era de 31,4 años, el sistema funcional mas frecuentemente afectado era el sensitivo (45,1%), el inicio fue monofocal en el 55,4% y el grado de discapacidad en la Expanded Disability Status Scale era de 2,1 puntos. La seroprevalencia del VEB fue del 99,3%. La reactivacion de la infeccion por VEB se relaciono con actividad clinica de EM en 10 pacientes (45,4%). Conclusiones. Actualmente, la prevalencia de la EM en la Region de Murcia es similar a la estimada en otras comunidades autonomas españolas. El estudio confirma la tendencia de incremento de prevalencia observada en las ultimas decadas.

Chicken ovalbumin upstream promoter-transcription factor I represses the transcriptional activity of the human muscle glycogen phosphorylase promoter in C2C12 cells.

The responsiveness of the 1.13 kb proximal human muscle glycogen phosphorylase (MGP) gene promoter to the chicken ovalbumin upstream promoter-transcription factor (COUP-TF) repressor, known to be ablated during muscle cell differentiation, was examined. Constitutive expression of COUP-TFI repressed the activity of the promoter in C2C12 muscle cells and sequential deletion analysis mapped the sensitive region between nucleotides -362 and -185, which included a putative consensus COUP-TF binding half-site at -198/-193. Mutation of this site abolished transcriptional response to COUP-TFI of the -362 construct. A -209/-180 probe bound in vitro to COUP-TFI and to protein extracts from proliferating but not fusing myoblasts. Thus, COUP-TF may be involved in repression of the human MGP gene promoter at the myoblast stage.

A glucose response element from the S. cerevisiae hexose transporter HXT1 gene is sensitive to glucose in human fibroblasts.

Glucose is an essential nutrient, and a regulator of gene expression in eukaryotic cells. Here, a comparative, function-based genomic approach has been used to identify glucose regulatory elements and transduction pathways common to both yeast and mammalian cells. We have isolated a region in the promoter of the Saccharomyces cerevisiae hexose transporter gene HXT1 that conferred glucose sensitivity in yeast, when located upstream of the minimal CYC1 promoter. This element contained binding motifs for Rgt1, a transcriptional modulator involved in the yeast glucose-induction pathway, that were sufficient to elicit glucose responsiveness. The HXT1 regulatory element was then fused to the minimal cytomegalovirus promoter (HXT1-MIN) and inserted into an adenovirus for delivery to human fibroblasts, where it exhibited glucose-dependent transcriptional activation. Glucose action was mimicked by fructose and unrelated to glucose 6-P content, whilst non-metabolizable glucose analogues showed no effect. Activation of AMP kinase by 5-aminoimidazole-4-carboxamide 1-beta-D-ribofuranosanide blocked glucose induction, revealing parallels with the yeast glucose-repressing pathway. In contrast, delivery of Rgt1 to fibroblasts did not modify HXT1-MIN responsiveness. Thus, elements of the S.cerevisiae HXT1 gene conserve glucose regulation in human fibroblasts equivalent to the metabolism-dependent, glucose-repressing pathway in yeast. These data suggest that the instructions carried within gene regulatory elements controlling nutrient regulation of gene expression have been conserved throughout evolution.

Trends in mortality due to motor vehicle traffic accident injuries between 1987 and 2011 in a Spanish region (Comunitat Valenciana).

OBJECTIVE: To analyse the time evolution of the rates of mortality due to motor vehicle traffic accidents (MVTA) injuries that occurred among the general population of Comunitat Valenciana between 1987 and 2011, as well as to identify trend changes by sex and age group. METHODS: An observational study of annual mortality trends between 1987 and 2011. We studied all deaths due to MVTA injuries that occurred during this period of time among the non-institutionalised population residing in Comunitat Valenciana (a Spanish Mediterranean region that had a population of 5,117,190 inhabitants in 2011). The rates of mortality due to MVTA injuries were calculated for each sex and year studied. These rates were standardised by age for the total population and for specific age groups using the direct method (age-standardised rate - ASR). Joinpoint regression models were used in order to detect significant trend changes. Additionally, the annual percentage change (APC) of the ASRs was calculated for each trend segment, which is reflected in statistically significant joinpoints. RESULTS: For all ages, ASRs decrease greatly in both men and women (70% decrease between 1990 and 2011). In 1990 and 2011, men have rates of 36.5 and 5.2 per 100,000 men/year, respectively. In the same years, women have rates of 8.0 and 0.9 per 100,000 women/year, respectively. This decrease reaches up to 90% in the age group 15-34 years in both men and women. ASR ratios for men and women increased over time for all ages: this ratio was 3.9 in 1987; 4.6 in 1990; and 5.8 in 2011. For both men and women, there is a first significant segment (p<0.05) with an increasing trend between 1987 and 1989-1990. After 1990, there are 3 segments with a significant decreasing APC (1990-1993, 1993-2005 and 2005-2011, in the case of men; and 1989-1996, 1999-2007 and 2007-2011, in the case of women). CONCLUSION: The risk of death due to motor vehicle traffic accidents injuries has decreased significantly, especially in the case of women, for the last 25 years in Comunitat Valenciana, mainly as of 2006. This may be a consequence of the road-safety measures that have been implemented in Spain and in Comunitat Valenciana since 2004. The economic crisis that this country has undergone since 2008 may have also been a contributing factor to this decrease. Despite the decrease, ASR ratios for men and women increased over time and it is still a high-risk cause of death among young men. It is thus important that the measures that helped decrease the risk of death are maintained and improved over time.

Laser scanning cytometry to quantify gene transfer efficiency and transcriptional activity of EGFP constructs.

Enhanced green fluorescent protein (EGFP) is the preferred reporter protein for real-time detection in individual cells, but its usefulness for gene expression quantification is limited by the sensitivity of standard detection techniques. We tested whether the unique feature of single-cell detection and quantification by laser scanning cytometry permits the evaluation of EGFP gene expression in monolayer cultures of kidney epithelial 293 and C2C12 muscle cells. Cells were transfected with plasmids expressing EGFP under the control of either the cytomegalovirus or muscle promoters: namely, muscle creatine kinase (MCK) and muscle glycogen phosphorylase (MGP). Cell monolayers were laser-scanned, fluorescence-imaged, and recorded. A population of fluorescence-emitting cells was discriminated, their contour area was defined, and the integrated fluorescence was estimated. These data were used to assess gene transfer efficiency in cells transfected with CMV-EGFP, which was higher in 293 than in C2C12 cells. Analysis of fluorescence intensity revealed that, as expected, CMV constructs were highly expressed in both cell types, whereas MCK and MGP constructs showed the highest transcriptional activity in C12C12 cells. In summary, we describe the utility of laser scanning cytometry for the automated estimation of gene transfer efficiency and transcriptional activity of EGFP constructs in cell monolayers.

Mouse PeP: a novel peroxisomal protein linked to myoblast differentiation and development.

The identification of several peroxisomal proteins in the past decade has deepened our understanding of the biology of peroxisomes and their involvement in human disorders. We report the cloning and expression pattern during the mouse development of a cDNA encoding a novel protein, named PeP, and show that its product is imported specifically to the peroxisome matrix in a variety of cell types. We also demonstrate that PeP is imported to the organelle through the PEX5 receptor pathway, which indicates that the C-terminal tripeptide SKI behaves as a type 1 peroxisomal targeting signal (PTS1). PeP expression is tightly regulated, as shown by Northern and in situ hybridization experiments. Thus during embryonic development in the mouse, PeP mRNA is detected almost exclusively in the skeletal muscle, whereas in adult mice, strong expression is also found in the heart and brain. In addition, PeP mRNA accumulation is induced after myoblast differentiation in vitro, when myotube formation is promoted. Sequence analysis reveals that PeP has no significant homology to any known protein, except for a short stretch of amino acids containing the fingerprint of the fibronectin type III superfamily, a domain present in proteins often related to molecular and cellular recognition and binding processes. Thus our data suggest a connection between the function of PeP and murine cell differentiation and development.

beta3-adrenoceptor agonist prevents alterations of muscle diacylglycerol and adipose tissue phospholipids induced by a cafeteria diet.

BACKGROUND: Insulin resistance induced by a high fat diet has been associated with alterations in lipid content and composition in skeletal muscle and adipose tissue. Administration of beta3-adrenoceptor (beta3-AR) agonists was recently reported to prevent insulin resistance induced by a high fat diet, such as the cafeteria diet. The objective of the present study was to determine whether a selective beta3-AR agonist (ZD7114) could prevent alterations of the lipid profile of skeletal muscle and adipose tissue lipids induced by a cafeteria diet. METHODS: Male Sprague-Dawley rats fed a cafeteria diet were treated orally with either the beta3-AR agonist ZD7114 (1 mg/kg per day) or the vehicle for 60 days. Rats fed a chow diet were used as a reference group. In addition to the determination of body weight and insulin plasma level, lipid content and fatty acid composition in gastronemius and in epididymal adipose tissue were measured by gas-liquid chromatography, at the end of the study. RESULTS: In addition to higher body weights and plasma insulin concentrations, rats fed a cafeteria diet had greater triacylglycerol (TAG) and diacylglycerol (DAG) accumulation in skeletal muscle, contrary to animals fed a chow diet. As expected, ZD7114 treatment prevented the excessive weight gain and hyperinsulinemia induced by the cafeteria diet. Furthermore, in ZD7114 treated rats, intramyocellular DAG levels were lower and the proportion of polyunsaturated fatty acids, particularly arachidonic acid, in adipose tissue phospholipids was higher than in animals fed a cafeteria diet. CONCLUSIONS: These results show that activation of the beta3-AR was able to prevent lipid alterations in muscle and adipose tissue associated with insulin resistance induced by the cafeteria diet. These changes in intramyocellular DAG levels and adipose tissue PL composition may contribute to the improved insulin sensitivity associated with beta3-AR activation.

Bloqueos tronculares de la extremidad inferior en pacientes críticos: A propósito de dos casos

We report two cases of anesthesia of sciaticpopliteal interne nerve, sciaticpopliteal external nerve and femoral nerve in critical ill patients under vascular surgery of lower limbs. One case is a patient diagnosed of recent pulmonary embolism and the other one diagnosed of recent myocardial infarction. The procedures were successful in both cases without complications. We recommend this procedure in critical patients undergoing surgery of lower extremities.

Profilaxis primaria de la fiebre reumática en la amigdalofaringitis aguda: estudio clínico, bacteriológico y terapéutico / Primary prophylaxis of rheumatic fever in acute pharyngotonsillitis: clinical, bacteriological, and therapeutical study

Se realizó un estudio clínico, bacteriológico y terapéutico a 200 niños con amigdalofaringitis aguda que asistieron al Cuerpo de Guardia del Hospital Pediátrico Marfán del municipio Plaza. Los pacientes eran vistos por el médico y se clasificaban por la clínica en tres grupos: I. clínicamente estreptocócicas, tratados con penicilina benzatínica (74 niños): II. Clínicamente estreptocócicas, tratados con otros antibacterianos (80 pacientes): III. Clínicamente no estreptocócicas, tratados sintomáticamente (46 niños). Todos fueron citados a reconsulta a la semana para valorar la evolución clínica y bacteriológica.La amigdalofaringitis clínicamente estreptocócica fue la forma clínica predominante y el estreptococo beta hemolítico del grupo A (E BH A) el microorganismo patógeno más frecuentemente aislado en los grupos I y II. Se logró negativizar el cultivo de E BH A en la mayoría de los que fueron positivos en el exudado inicialy se corroboró la efectividad del uso de la penicilina benzatínica en el tratamiento de esta enfermedad