Cambios inducidos en DKK1 en pacientes con artritis reumatoide que inician tratamiento con terapia biológica / Changes induced by DKK1 in rheumatoid arthritis patients who commence biologic therapy treatment

Introducción: El objetivo del estudio es evaluar la relación entre la carga inflamatoria, el riesgo cardiovascular y el metabolismo óseo en pacientes con artritis reumatoide que inician tratamiento con terapia biológica. Pacientes y métodos: Se trata de un estudio de cohortes prospectivo realizado en pacientes con diagnóstico de artritis reumatoide (AR) activa evaluados en la Unidad de Reumatología y que inician terapia biológica. Los pacientes serán seleccionados de forma consecutiva. Presentamos los datos preliminares de 14 pacientes. Resultados: Encontramos una reducción en las concentraciones de Dickkopf-1 (DKK1) tras el inicio de la terapia biológica (basal: 53,12±60,43 pg/ml vs. 6 meses 23,2±13,5 pg/ml, p=0,307) pero no se alcanzó la significación estadística. Se encontraron cambios en los marcadores de remodelado con aumento en los niveles de osteocalcina y CTX que no alcanzó la significación estadística. Conclusiones: En pacientes con AR activa tratados con terapia biológica hemos observado un descenso no significativo de las concentraciones séricas de DKK1. La ampliación tanto de los sujetos de estudio como de las determinaciones bioquímicas pendientes nos permitirán en un futuro próximo establecer de forma más precisa esta asociación, así como la relación entre DKK1, remodelado óseo, terapia biológica y enfermedad cardiovascular en pacientes con AR (AU)

Introduction: The aim of this study is to assess the relationship among inflammatory charge, cardiovascular risk and bone metabolism in patients with rheumatoid arthritis initiating biological therapy treatment. Patients and methods: This is a prospective cohort study conducted in patients diagnosed with active rheumatoid arthritis (RA) assessed in the Rheumatology Unit and initiating biological therapy. Patients will be selected consecutively, with preliminary data on 14 patients. We present preliminary data from 14 patients. Results: Reduced Dickkopf-1 (DKK1) concentrations after commencing biological therapy were detected (baseline: 53.12±60.43 pg/ml vs 6 months 13.5±23.2 pg/ml, p=0.307) but without statistical significance. Changes were found in markers for bone remodeling with increased osteocalcin levels and CTX which were not statistically significant either. Conclusions: We observed a nonsignificant decrease in DKK1 serum in patients with active RA treated with biologic therapy. Expanding the scope of study subjects and pending biochemical determinations will allow us, in the near future, to establish more precisely this link and the relationship of DKK1, bone remodeling, biological therapy and cardiovascular disease in RA patients (AU)

A multistrategic approach in the development of sourdough bread targeted towards blood pressure reduction.

Rising prevalence of hypertension is pushing food industry towards the development of innovative food products with antihypertensive effects. The aim was to study the effect of reduced sodium content and 21% addition of wholemeal wheat sourdough (produced by Lactobacillus brevis CECT 8183 and protease) on proximate composition, γ-aminobutyric acid (GABA) and peptide content of wheat bread. Angiotensin converting enzyme I (ACE) inhibitory and antioxidant activities were also evaluated. Sodium replacement by potassium salt did not affect chemical composition and biological activities of bread. In contrast, GABA and peptides <3 kDa contents in sourdough bread (SDB) were 7 and 3 times higher, respectively, than the observed in control. ACE inhibitory and antioxidant activities of the peptide fraction < 3 kDa from SDB was 1.7 and 2.6-3.0 times higher than control. Therefore, the combination of reduced sodium content with enriched concentrations of bioactive compounds in bread making may provide interesting perspectives for development of innovative breads towards blood pressure reduction.

A B-cell epigenetic signature defines three biologic subgroups of chronic lymphocytic leukemia with clinical impact.

Prospective identification of patients with chronic lymphocytic leukemia (CLL) destined to progress would greatly facilitate their clinical management. Recently, whole-genome DNA methylation analyses identified three clinicobiologic CLL subgroups with an epigenetic signature related to different normal B-cell counterparts. Here, we developed a clinically applicable method to identify these subgroups and to study their clinical relevance. Using a support vector machine approach, we built a prediction model using five epigenetic biomarkers that was able to classify CLL patients accurately into the three subgroups, namely naive B-cell-like, intermediate and memory B-cell-like CLL. DNA methylation was quantified by highly reproducible bisulfite pyrosequencing assays in two independent CLL series. In the initial series (n=211), the three subgroups showed differential levels of IGHV (immunoglobulin heavy-chain locus) mutation (P<0.001) and VH usage (P<0.03), as well as different clinical features and outcome in terms of time to first treatment (TTT) and overall survival (P<0.001). A multivariate Cox model showed that epigenetic classification was the strongest predictor of TTT (P<0.001) along with Binet stage (P<0.001). These findings were corroborated in a validation series (n=97). In this study, we developed a simple and robust method using epigenetic biomarkers to categorize CLLs into three subgroups with different clinicobiologic features and outcome.

High-pressure improves enzymatic proteolysis and the release of peptides with angiotensin I converting enzyme inhibitory and antioxidant activities from lentil proteins.

Angiotensin I converting enzyme (ACE) inhibitory and antioxidant peptides are receiving attention due to their beneficial effects in the prevention/treatment of hypertension. The objective was to explore the effect of high hydrostatic pressure (HP) on proteolysis by different proteases and the release of bioactive peptides from lentil proteins. Pressurisation (100-300 MPa) enhanced the hydrolytic efficiency of Protamex, Savinase and Corolase 7089 compared to Alcalase. Proteolysis at 300 MPa led to a complete degradation of lentil proteins and increased peptide (<3 kDa) concentration by all enzymes. Proteolysis at 300 MPa by Savinase gave rise to lentil hydrolysates (S300) with the highest ACE-inhibitory and antioxidant activities that were retained upon in vitro gastrointestinal digestion. The peptides responsible for the multifunctional properties of S300 hydrolysate were identified as different fragments from storage proteins and the allergen Len c 1. These results support the potential of HP as a technology for the cost-effective production of bioactive peptides from lentil proteins during enzymatic proteolysis.

Ochratoxin A kinetics: a review of analytical methods and studies in rat model.

Ochratoxin A (OTA) is a thermostable mycotoxin that contaminates a great variety of foodstuffs. It is nephrotoxic in all of the mammalian species tested, the pig being the most sensitive one; among rodents, rats are the most susceptible to OTA carcinogenicity. Kinetics, by studying the absorption, distribution, metabolism and excretion of xenobiotics, is an important tool in the extrapolation of animal toxicity data for human risk assessment. The most important kinetic studies performed with OTA in rats are reviewed, together with the different methods used for OTA quantification in biological matrices. Twelve studies in Wistar, Sprague-Dawley or F344 rats, using radiolabeled OTA or TLC, HPLC-FLD or LC/MS have been summarized. Very often methods validated for food have been directly applied to tissues. Strain, sex and age differences have been detected but the interpretation is difficult due to the different experimental conditions, and the connection of the several factors that may account for these differences.

Allergy to all mammalian Bovidae proteins but cow's milk in a child

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Extruded flaxseed meal enhances the nutritional quality of cereal-based products.

Human consumption of flaxseed is increasing due to its health benefit properties and extrusion processes can enhance its nutritional quality. Extruded flaxseed meal (EFM) obtained in a pilot plant was characterized and incorporated in flour mixes and cereal-based bars to demonstrate its nutritious usefulness. Amino acid content was not affected by extrusion and, despite lysine was the limitating amino acid, the chemical score (CS) was 83 %. Thiamin and riboflavin decreased slightly as consequence of extrusion, phytic acid did not change and trypsin inhibitor activity was undetectable. Proximate composition and nutritional quality determined by biological and chemical indexes were compared among EFM, flour mixes (FM) and cereal bars (CB). They presented high protein levels (26, 20 and 17 %, respectively), good biological value (BV) (80, 79 and 65, respectively), acceptable true protein digestibility (TD) (73, 79 and 78, respectively), and high dietary fiber (33, 20.5 and 18 %, respectively). The ratio of ω6:ω3 for CB was within the WHO/FAO recommendations. These results open a new venue for the usefulsess of nutritious/healthy extruded flaxseed flours into ready-to-eat cereal-based products with improved nutritional quality.

An approach to the toxicity and toxicokinetics of aflatoxin B1 and ochratoxin A after simultaneous oral administration to fasted F344 rats.

Humans are exposed to the hepatotoxic aflatoxin B1 (AFB1) and nephrotoxic ochratoxin A (OTA) through diet. However, kinetic and toxicological data after their co-administration are scarce. In this study, a single oral dose of AFB1 (0.25 mg/kg bw)+OTA (0.5 mg/kgbw) was administered to fasted F344 rats. Blood, liver and kidney were harvested at different timepoints for mycotoxins quantification, relative weight calculation, clinical biochemistry and histopathology analysis. Toxicity parameters pointed to acute toxicity in liver due to AFB1. No remarkable toxicity was observed in kidneys or immunological organs. Maximum observed concentrations in plasma (Cmax) were at 10 min and 2 h for AFB1 and OTA, respectively. AFB1 plasma concentration could indicate a rapid absorption/ metabolism of the mycotoxin; and AFB1 liver and kidney concentrations were lower than LOQ and LOD, respectively. For OTA, Cmax was 4326.2 µg/L in plasma. In kidney and liver Cmax was reached at 8 h and concentrations were very similar between both organs at all timepoints. Due to the low levels of AFB1, the effect of OTA on AFB1 kinetics could not be assessed. However, AFB1 seems not to affect OTA kinetics, as its profile seems very similar to kinetic studies performed only with OTA in similar conditions.

Conocimientos y actitudes de la población ante la gripe A (H1N1) / Knowledge and attitudes of population on influenza A (H1N1)

Introducción. La pandemia de gripe A ha supuesto un reto para los sistemas sanitarios mundiales, conllevando una considerable alarma social. No tenemos la seguridad de que los mensajes de las autoridades sanitarias hayan llegado adecuadamente a la población. Objetivo: describir el grado de conocimiento y las actitudes de la población sobre prevención y tratamiento de la gripe A. Métodos. Estudio descriptivo transversal por reclutamiento de usuarios mayores de edad que acuden al Centro de Salud mediante encuesta autocumplimentada elaborada ad hoc. Resultados. Doscientas seis encuestas (60% mujeres), edad media 49,8±17,4 años. El 33,5% se consideraban pertenecientes a grupos de riesgo frente a la gripe A. El 29,13% se había vacunado el año anterior frente a la gripe, por 27,8% el año actual (7,14% frente a gripe A). Los conocimientos correctos sobre síntomas, tratamiento y necesidad de receta para conseguir antivirales fueron del 61%, 32,7% y 70,3%, respectivamente. El 40,1% se consideraba poco o nada informado, obteniendo la información fundamentalmente de la televisión (75,7%). El 37,81% se consideraba bastante o muy preocupado ante la enfermedad, y el 37,24% consideraba que esta era bastante o muy grave. Las actitudes ante el padecimiento de un cuadro catarral no cambiaron respecto a la pauta habitual. Conclusiones. La cobertura vacunal es baja, especialmente ante la gripe A. La población se considera poco informada sobre síntomas y tratamiento de la gripe A. La mayor parte de la información se obtiene de medios no oficiales (televisión). El grado de preocupación y la percepción de gravedad se encuentran en niveles medios. La pandemia no ha alterado las costumbres de la población (AU)

Introduction. The influenza A (H1N1) outbreak has been a challenge for world health systems; it has provoked significant social alarm. We do not know if the health authorities advertisements have been understood by the population. Objective. To describe the knowledge and attitudes of the population on the prevention and treatment of influenza A (H1N1). Methods. Descriptive, cross-sectional study of a population who attended their Health Centre. A self-completed questionnaire was given. Results. There were 206 responses (60% women), with a mean age 49.8±17.4 years old, of which 33.5% thought they belonged to a risk group for A (H1N1)-flu. A total of 29.13% had been immunized against usual flu last season, and 27.8% this season (7.14% against A-flu). Proper knowledge about symptoms, treatment and the need of a prescription for antivirals were 61%, 32.7% and 70.3%, respectively. There was 40.1% who said they had little or no information, getting their information mainly from television (75.7%). A total of 37.81% were quite worried or very worried about the illness, and 37.24% thought it was severe or very severe. The attitudes relating to suffering the illness were similar to seasonal flu. Conclusions. Flu vaccination coverage (especially A-flu) is low. Knowledge of A (H1N1) flu symptoms and treatment was low, and a significant percentage of the population said they had little or no information. The information is mostly taken from unofficial channels (television). Concern about the illness is not great, but there is a perception of the severity. A-flu pandemic has not changed the attitudes of the population (AU)

Effects of fasting and gender on ochratoxin A toxicokinetics in F344 rats.

Ochratoxin A (OTA) is a mycotoxin that causes renal tumors in rats, particularly in males. In previous kinetic studies performed in fed conditions (Vettorazzi et al., 2008), mature F344 male rats presented a significantly lower OTA bioavailability than females and young animals. The objective of the present study was to evaluate two factors which could explain this different kinetic profile: the presence of food and the male-specific protein alpha-2u-globulin. Therefore, a 24h kinetic study has been performed in rats under fasting conditions. Food ingestion has been controlled in both sexes during two months. The presence of alpha-2u-globulin in the urine has been analyzed with SDS-gradient mini-gel electrophoresis. Fasting tends to increase the maximum OTA plasma concentrations and the rate of absorption. The relative bioavailability is significantly increased under fasting conditions only in males. Mature males consumed a higher amount of food but, as the OTA dose administered, it was proportional to body weight. The reason why the OTA bioavailability is more affected in presence of food only in males is unclear. Several possibilities, such as differences in gastric emptying, OTA-food interactions and the involvement of alpha-2u-globulin are discussed.

Comparison between capillary electrophoresis and high performance liquid chromatography for the study of the occurrence of patulin in apple juice intended for infants.

Apple juice samples intended for infants purchased in Navarra (Spain) have been analyzed for PAT occurrence. Two capillary electrophoresis methods, based on a MEKC and a CEC system, and an HPLC method were evaluated for the aforementioned study. The CEC system gave less satisfying separations and several practical problems, so samples have been analyzed by MEKC and HPLC. Both methods have been comparable in terms of recovery, precision, limits of detection, volume of organic solvents used and adequate selectivity with regard to PAT and HMF. The analysis time in HPLC has been slightly lower than in the MEKC methodology. The PAT levels obtained in apple juice by both validated methods showed a strong correlation (p<0.001). Therefore, both methodologies are useful for the accurate quantification of patulin in this matrix. The PAT levels obtained in the 20 infant apple juices samples were in a range between

A different kinetic profile of ochratoxin A in mature male rats.

Ochratoxin A (OTA) is a mycotoxin that causes renal tumors in rodents, particularly in male rats. The present work explored the impact of gender and age on OTA toxicokinetics in F344 rats after a single oral dose (0.5mg/kg b.w.). OTA plasma concentrations were analysed with a validated HPLC-FLD method and a population approach (NONMEM VI) was used to perform the kinetic analysis and the one year exposure simulation (0.21 mg/kg daily). Maximum observed OTA concentration (CMAX(obs)) was at 2h in all groups except in mature females (6h). Mature females reached higher CMAX(obs) than males of the same age. Apparent volume of distribution, but not apparent total plasma clearance, increased significantly with body weight (P<0.01) resulting in the following values for the terminal plasma half life (h) in males: 219 (young), 264 (matures) and females: 191 (young), 205 (matures). In addition mature males showed a significant lower relative bioavailability. The simulation showed similar plasma concentrations in males and females after two-months. Thus, toxicokinetic does not seem to explain sex-differences in toxicity in long-term studies. However, the age and weight should be taken into account in short-term toxicological studies if sex-differences are studied.

Influence of fermentation conditions on glucosinolates, ascorbigen, and ascorbic acid content in white cabbage (Brassica oleracea var. capitata cv. Taler) cultivated in different seasons.

The content of glucosinolates (GLS), ascorbigen, and ascorbic acid in white cabbage (Brassica oleracea var. capitata cv. Taler) cultivated in different seasons (summer and winter) was determined, before and after spontaneous and starter-induced fermentation. Different salt concentrations (0.5% NaCl or 1.5% NaCl) were used for sauerkraut production. Glucoiberin, sinigrin, and glucobrassicin were dominating in raw white cabbage cultivated either in winter or summer seasons. Ascorbigen precursor, glucobrassicin, was found higher in cabbage cultivated in winter (2.54 micromol/g dw) than those grown in summer (1.83 micromol/g dw). Cabbage fermented for 7 d was found to contain only traces of some GLS irrespective of the fermentation conditions used. Ascorbigen synthesis occurred during white cabbage fermentation. Brining cabbage at low salt concentration (0.5% NaCl) improved ascorbigen content in sauerkraut after 7 d of fermentation at 25 degrees C. The highest ascorbigen concentration was observed in low-sodium (0.5% NaCl) sauerkraut produced from cabbage cultivated in winter submitted to either natural (109.0 micromol/100 g dw) or starter-induced fermentation (108.3 and 104.6 micromol/100 g dw in cabbages fermented by L. plantarum and L. mesenteroides, respectively). Ascorbic acid content was found higher in cabbage cultivated in summer and fermentation process led to significant reductions. Therefore, the selection of cabbages with high glucobrassicin content and the production of low-sodium sauerkrauts may provide enhanced health benefits towards prevention of chronic diseases.

A simple chemical method reduces ochratoxin A in contaminated cocoa shells.

Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus and Penicillium species, which contaminates cocoa among other food commodities. It has been previously demonstrated that the toxin is concentrated in cocoa shells. The aim of this study was to assay a simple chemical method for ochratoxin A reduction from naturally contaminated cocoa shells. In order to determine the efficiency of the method, a high-performance liquid chromatography method with fluorescence detection was set up beforehand and validated. Ochratoxin A was extracted from cocoa shells with methanol-3% sodium bicarbonate solution and then purified with immunoaffinity columns. The recovery attained was 88.7% (relative standard deviation = 6.36%) and the limits of detection and quantification were 0.06 and 0.2 kg/kg, respectively. For decontamination experiments, the solvent extractor ASE 200 was used. First, aqueous solutions of 2% sodium bicarbonate and potassium carbonate were compared under the same conditions (1,500 lb/in2 at 40 degrees C for 10 min). Higher ochratoxin A reduction was obtained with potassium carbonate (83 versus 27%). Then, this salt was used under different conditions of pressure, temperature, and time. The greatest ochratoxin A reduction was achieved with an aqueous potassium carbonate solution (2%), at 1,000 lb/in2 at 90 degrees C for 10 min. This method could probably be applicable to the cocoa industry because it is fast and relatively economic. From the point of view of human health, the use of potassium carbonate, partially eliminated by rinsing the sample with water, does not likely represent a risk for human health.

Development and validation of a microemulsion electrokinetic chromatography method for patulin quantification in commercial apple juice.

A microemulsion electrokinetic chromatography (MEECK) method for patulin (PAT) quantification in apple juice samples has been developed. The effects of several important factors such as co-surfactant type, concentration of surfactant, acetonitrile percentage in the microemulsion, and running voltage and temperature were investigated to determine the optimum conditions. They resulted to be: a background electrolyte (BGE) composed of 25mM of sodium tetraborate, SDS (2.16%w/w), ethanol (6.49%w/w), n-octanol (0.82%w/w) and 2%v/v acetonitrile; applied voltage of +15kV; and a capillary temperature of 35 degrees C. PAT was detected at 276nm. Quantification and detection limits (LOQ and LOD) in apple juice samples were 8.0microgL(-1) and 3.2microgL(-1), respectively. Patulin was extracted from apple juice using ethyl acetate with a mean recovery value of 75.3% (RSD=4.5). This method was applied to the measurement of patulin in twenty commercial apple juice samples obtained from different Danish supermarkets. The PAT apple juice mean and median levels obtained were 35.9 and 10.9microgL(-1), respectively. The comparison with a previously validated micellar electrokinetic chromatography (MEKC) method for PAT analysis showed the suitability of using MEEKC for this mycotoxin analysis. However, the expectations of obtaining a higher efficiency and thus lower limits of detection and quantitation when using MEEKC were not met.

Impact of genetic abnormalities on survival after allogeneic hematopoietic stem cell transplantation in multiple myeloma.

We analyzed the prognostic impact of the most frequent genetic abnormalities detected by fluorescence in situ hybridization in 101 patients with multiple myeloma, who underwent allogeneic hematopoietic stem cell transplantation (HSCT) after melphalan/fludarabine-based reduced conditioning. The incidences of abnormalities in the present analysis were as follows: del(13q14) (61%), t(11;14)(q13;q32) (14%), t(4;14)(p16.3;q32) (19%), MYC-gain gains (8q24) (21%), del(17p13) (16%) and t(14;16)(q32;q23) (5%). None of the patients had t(6;14)(p25;q32). The overall complete remission (CR) rate was 50% with no differences between the genetic abnormalities except for patients with del(17p13) who achieved less CR (7 vs 56%; P=0.001). Univariate analysis revealed a higher relapse rate in patients aged >50 years (P=0.002), patients with del(13q14) (P=0.006) and patients with del(17p13) (P=0.003). In multivariate analyses, only del(13q14) (HR: 2.34, P=0.03) and del(17p13) (HR: 2.24; P=0.04) significantly influenced the incidence of relapse, whereas for event-free survival, only age (HR 2.8; P=0.01) and del(17p13) (HR: 2.05; P=0.03) retained their negative prognostic value. These data show that del(17p13) is a negative prognostic factor for achieving CR as well as for event-free survival after HSCT. Translocation t(4;14) might be overcome by allogeneic HSCT, which will have implication for risk-adapted strategies.

OTA-producing fungi isolated from stored cocoa beans.

AIMS: The aim of this study was to identify fungal populations in unroasted cocoa beans stored in Spain in order to evaluate the ochratoxin A (OTA)-production ability of certain Aspergillus isolates. METHODS AND RESULTS: Twenty batches of cocoa beans from different origins and with different OTA content were selected for this study. Three Aspergillus carbonarius and 13 Aspergillus niger aggregate strains isolated from these cocoa bean samples were selected to evaluate their OTA synthesis ability, being the only A. carbonarius isolates which are OTA producers [