Digestibility of Quinoa (Chenopodium quinoa Willd.) Protein Concentrate and Its Potential to Inhibit Lipid Peroxidation in the Zebrafish Larvae Model.

Quinoa protein concentrate (QPC) was extracted and digested under in vitro gastrointestinal conditions. The protein content of QPC was in the range between 52.40 and 65.01% depending on the assay used. Quinoa proteins were almost completely hydrolyzed by pepsin at pH of 1.2, 2.0, and 3.2. At high pH, only partial hydrolysis was observed. During the duodenal phase, no intact proteins were visible, indicating their susceptibility to the in vitro simulated digestive conditions. Zebrafish larvae model was used to evaluate the in vivo ability of gastrointestinal digests to inhibit lipid peroxidation. Gastric digestion at pH 1.2 showed the highest lipid peroxidation inhibition percentage (75.15%). The lipid peroxidation activity increased after the duodenal phase. The digest obtained at the end of the digestive process showed an inhibition percentage of 82.10%, comparable to that showed when using BHT as positive control (87.13%).

Is targeted removal a suitable means for tuberculosis control in wild boar?

We assessed the suitability of targeted removal as a means for tuberculosis (TB) control on an intensely managed Eurasian wild boar (Sus scrofa) hunting estate. The 60km2 large study area included one capture (treatment) site, one control site, and one release site. Each site was fenced. In the summers of 2012, 2013 and 2014, 929 wild boar were live-captured on the treatment site. All wild boar were micro-chipped and tested using an animal side lateral flow test immediately after capture in order to detect antibodies to the Mycobacterium tuberculosis complex (MTC). The wild boar were released according to their TB status: Seropositive individuals onto the release site (hunted after summer), and seronegative individuals back onto the treatment site. The annual summer seroprevalence of antibodies to the MTC declined significantly in live-captured wild boar piglets from the treatment site, from 44% in 2012 to 27% in 2013 (a reduction of 39%). However, no significant further reduction was recorded in 2014, during the third capture season. Fall-winter MTC infection prevalence was calculated on the basis of the culture results obtained for hunter-harvested wild boar. No significant changes between hunting seasons were recorded on either the treatment site or the control site, and prevalence trends over time were similar on both sites. The fall-winter MTC infection prevalence on the release site increased significantly from 40% in 2011-2012 to 64% in 2012-2013 and 2013-2014 (60% increase). Recaptures indicated a persistently high infection pressure. This experiment, the first attempt to control TB in wild boar through targeted removal, failed to reduce TB prevalence when compared to the control site. However, it generated valuable knowledge on infection pressure and on the consequences of translocating TB-infected wild boar.

Antimicrobial resistance in faecal Escherichia coli isolates from farmed red deer and wild small mammals. Detection of a multiresistant E. coli producing extended-spectrum beta-lactamase.

Eighty-nine Escherichia coli isolates recovered from faeces of red deer and small mammals, cohabiting the same area, were analyzed to determine the prevalence and mechanisms of antimicrobial resistance and molecular typing. Antimicrobial resistance was detected in 6.7% of isolates, with resistances to tetracycline and quinolones being the most common. An E. coli strain carrying blaCTX-M-1 as well as other antibiotic resistant genes included in an unusual class 1 integron (Intl1-dfrA16-blaPSE-1-aadA2-cmlA1-aadA1-qacH-IS440-sul3-orf1-mef(B)Δ-IS26) was isolated from a deer. The blaCTX-M-1 gene was transferred by conjugation and transconjugants also acquired an IncN plasmid. This strain was typed as ST224, which seems to be well adapted to both clinical and environmental settings. The phylogenetic distribution of the 89 strains varied depending on the animal host. This work reveals low antimicrobial resistance levels among faecal E. coli from wild mammals, which reflects a lower selective pressure affecting these bacteria, compared to livestock. However, it is remarkable the detection of a multi-resistant ESBL-E. coli with an integron carrying clinically relevant antibiotic-resistance genes, which can contribute to the dissemination of resistance determinants among different ecosystems.

Testing Eurasian wild boar piglets for serum antibodies against Mycobacterium bovis.

Animal tuberculosis (TB) caused by infection with Mycobacterium bovis and closely related members of the M. tuberculosis complex (MTC), is often reported in the Eurasian wild boar (Sus scrofa). Tests detecting antibodies against MTC antigens are valuable tools for TB monitoring and control in suids. However, only limited knowledge exists on serology test performance in 2-6 month-old piglets. In this age-class, recent infections might cause lower antibody levels and lower test sensitivity. We examined 126 wild boar piglets from a TB-endemic site using 6 antibody detection tests in order to assess test performance. Bacterial culture (n=53) yielded a M. bovis infection prevalence of 33.9%, while serum antibody prevalence estimated by different tests ranged from 19% to 38%, reaching sensitivities between 15.4% and 46.2% for plate ELISAs and between 61.5% and 69.2% for rapid immunochromatographic tests based on dual path platform (DPP) technology. The Cohen kappa coefficient of agreement between DPP WTB (Wildlife TB) assay and culture results was moderate (0.45) and all other serological tests used had poor to fair agreements. This survey revealed the ability of several tests for detecting serum antibodies against the MTC antigens in 2-6 month-old naturally infected wild boar piglets. The best performance was demonstrated for DPP tests. The results confirmed our initial hypothesis of a lower sensitivity of serology for detecting M. bovis-infected piglets, as compared to older wild boar. Certain tests, notably the rapid animal-side tests, can contribute to TB control strategies by enabling the setup of test and cull schemes or improving pre-movement testing. However, sub-optimal test performance in piglets as compared to that in older wild boar should be taken into account.

Coxiella burnetii Shedding by Farmed Red Deer (Cervus elaphus).

Wildlife and notably deer species--due to the increasing relevance of deer farming worldwide--may contribute to the maintenance of Coxiella burnetii, the causal agent of Q fever. Currently, there are no precedents linking exposure to deer species with human Q fever cases. However, a human case of Q fever was recently diagnosed in a red deer (Cervus elaphus) farm, which led us to investigate whether deer could be a source for environmental contamination with C. burnetii and ascertain the implication of C. burnetii in reproductive failure in the farm. Blood serum and vaginal swabs were collected from hinds either experiencing or not reproductive failure and tested to detect the presence of antibodies and DNA, respectively, of C. burnetii, Chlamydia abortus, Neospora caninum and Toxoplasma gondii. Serology and PCR results suggest C. burnetii was the primary cause of the reproductive failure. We identified vaginal shedding of C. burnetii in hinds, confirming red deer as a source of Q fever zoonotic infection.

Temporal trend of tuberculosis in wild ungulates from Mediterranean Spain.

Tuberculosis (TB), a chronic disease caused by infection with the Mycobacterium tuberculosis complex, is endemic in wild boar (Sus scrofa) and red deer (Cervus elaphus) in south-central Spain. Understanding the temporal dynamics of this chronic infection requires long time series data collection over large areas. The aim of this paper was to identify the determinants of TB prevalence and severity in both species in Ciudad Real province, Spain, from 2000 to 2012. Study variables included management, population dynamics, and a range of geographical and climatological factors. The prevalence of TB in wild boar increased from 50% to 63% since the study commenced. This may be due to an increased hunting bag (a proxy for population abundance), which was correlated with TB infection rates. Low rainfall (a stochastic factor) was associated with higher individual risk of TB presence and progression, resulting in an increased proportion of severe cases of wild boar TB in dry years. This was probably a result of increased food restriction leading to a higher susceptibility to TB. In contrast, red deer TB showed an apparent stable trend, which may be a consequence of the species' higher and stable population size. Hunting management, characterized by fencing, was associated with a higher risk of TB in both wild boar and red deer, suggesting that intensive hunting management may have contributed to exacerbated TB figures. This difference was more marked in red deer than in wild boar, probably because fencing imposes less restriction on movement, population mixing and TB spread to wild boar than to deer. Our findings on TB dynamics are fundamental for assessing the impact of future disease-control actions (e.g. field vaccination). Moreover, such control plans must operate in the long term and cover large areas.

Infestaciones por garrapatas: a propósito de un caso / Tick infestation: report of a case

Las garrapatas son parásitos hematófagos que afectan la piel de muchos animales vertebrados, transmitiéndose al ser humano por contacto directo con animales infestados. La infestación por garrapata se manifiesta como una mácula, pápula o placa eritematosa y pruriginosa, que aparece 1 a 3 días después de la exposición con el parásito. Además, puede estar asociada con infección local secundaria en el sitio de la mordedura, granuloma a cuerpo extraño, parálisis ascendente e incluso anafilaxia. Estos parásitos también pueden transmitir agentes patógenos capaces de producir enfermedades en el ser humano como bacterias, virus y protozoos. La garrapata debe ser extirpada, teniendo especial precaución de no comprimir demasiado el cuerpo del parásito durante el procedimiento. Reportamos el caso de un hombre de 63 años que presentó una placa eritematosa y pruriginosa en la espalda, cuya imagen dermatoscópica reveló la presencia de una garrapata.

Ticks are blood-sucking, opportunistic parasites that can attach to the skin of a variety of vertebrate hosts. Humans become infested by coming into close contact with infested animals. Tick infestation may present as erythematous pruritic macule, papule, or finely elevated plaque within 1­3 days following tick exposure. Also tick bites may be associated with local, secondary infection at the bite site, foreign body granuloma, ascending paralysis, and even anaphylaxis. In addition, these parasites may transmit a number of different pathogens that cause human diseases including bacteria, viruses, and protozoan. Ticks may be removed, although the body of the tick should not be squeezed during the removal process. We report a 63-year old man who presented with an itchy erythematous plaque on the back, which, upon closer inspection, revealed a partially embedded tick.

Potential use of vanadium compounds in therapeutics.

Vanadium is a trace element present in practically all cells in plants and animals. While the essentiality of vanadium for human beings remains to be well established, vanadium has become an increasingly important environmental metal. Vanadium compounds exert a variety of biological activities and responses. At pharmacological doses, vanadium compounds display relevant biological actions such as insulin and growth factor mimetic or enhancing effects, as well as osteogenic and cardioprotective activity. On the other hand, depending on the nature of compounds and their concentrations, toxicological actions and adverse side effects may also be shown. Nevertheless, the toxic effects may be useful to develop new antitumoral drugs. In this review, the authors summarize current knowledge and new advances on in vitro and in vivo effects of inorganic and organically-chelated vanadium compounds. The effects of vanadium derivatives on some cellular signaling pathways related to different diseases are compiled. In particular, the pathways relevant to the insulin mimetic, osteogenic, cadioprotective and antitumoral actions of vanadium compounds have been comprehensively reviewed. The knowledge of these intracellular signaling pathways may facilitate the rational design of new vanadium compounds with promising therapeutic applications as well as the understanding of secondary side effects derived from the use of vanadium as a therapeutic agent.

Cytotoxicity of a vanadyl(IV) complex with a multidentate oxygen donor in osteoblast cell lines in culture.

Strong chelating ligands as oxodiacetate (oda) are model systems to study the process of metal trapping by living organisms. Vanadium compounds display interesting biological and pharmacological actions. In vertebrates, vanadium is stored mainly in bones. In the present study we report the effects of the complex of oda with vanadyl(IV) cation, VO(oda), on two osteoblast cell lines, one normal (MC3T3E1) and the other tumoral (UMR106). VO(oda) exerted cytotoxic actions in osteoblasts as it was determined through a dose-dependent decrease in cell proliferation, and morphological and actin alterations. The putative mechanisms underlying VO(oda) deleterious effects were also investigated. The complex increased the level of ROS which correlated with a decreased in GSH/GSSG ratio. Besides, VO(oda) induced a dissipation of the mitochondria membrane potential (MMP) and promoted an increase in ERK cascade phosphorylation, which is involved in the regulation of cellular death and survival. All the effects were more pronounced in MC3T3-E1 than in UMR106 cells. ERK activation was inhibited by PD98059, Wortmanin and the ROS scavenger NAC (N-acetyl cysteine). These results suggest that VO(oda) stimulated ERKs phosphorilation by induction of free radicals involving kinases upstream of ERK pathway. The inhibitory effect of the complex on cell proliferation was partially reversed in both cell lines by NAC. Moreover, PD98059 and Wortmanin also partially reversed the inhibition of cell proliferation in the tumoral osteoblasts. The use of specific inhibitors and ROS scavengers suggested the involvement of oxidative stress, MMP alterations and ERK pathway in the apoptotic actions of this complex.

Optimising the use of computed radiography in pediatric chest imaging.

The objective of this study was to analyze image quality of chest examinations in pediatric patients using computed radiography (CR) obtained with a wide range of doses to suggest the appropriate parameters for optimal image quality. A sample of 240 chest images in four age ranges was randomly selected from the examinations performed during 2004. Images were obtained using a CR system and were evaluated independently by three radiologists. Each image was scored using criteria proposed by the European Guidelines on Quality Criteria in Pediatrics. Mean global scoring and scoring of individual criteria more sensitive to noise were used to evaluate image quality. Agfa dose level (DL) was in the range 1.20 to 2.85. It was found that there was not significant correlation (R < 0.5) between image quality and DL for any of the age ranges for either global score or for individual criteria more related to noise. The mean value of DL was in the ranges 1.9-2.1 for the four age bands. From this study, a DL value of 1.6 is proposed for pediatric CR chest imaging. This could yield a reduction of approximately a factor of 2.5 in mean patient entrance surface doses.

Continuous thermal hydrolysis and energy integration in sludge anaerobic digestion plants.

A thermal hydrolysis pilot plant with direct steam injection heating was designed and constructed. In a first period the equipment was operated in batch to verify the effect of sludge type, pressure and temperature, residence time and solids concentration. Optimal operation conditions were reached for secondary sludge at 170 degrees C, 7 bar and 30 minutes residence time, obtaining a disintegration factor higher than 10, methane production increase by 50% and easy centrifugation In a second period the pilot plant was operated working with continuous feed, testing the efficiency by using two continuous anaerobic digester operating in the mesophilic and thermophilic range. Working at 12 days residence time, biogas production increases by 40-50%. Integrating the energy transfer it is possible to design a self-sufficient system that takes advantage of this methane increase to produce 40% more electric energy.

Spectroscopic characterization of a VO2+ complex of oxodiacetic acid and its bioactivity on osteoblast-like cells in culture.

The oxovanadium(IV) complex of oxodiacetic acid (H2oda) of stoichiometry [VO(oda)(H2O)2], which presents an unprecedented tridentate OOO coordination, was thoroughly characterized by infrared, Raman, electronic, and electron paramagnetic resonance spectroscopies. The biological activity of the complex on the cell proliferation and differentiation was tested on osteoblast-like cells (MC3T3E1 osteoblastic mouse calvaria-derived cells and UMR106 rat osteosarcoma-derived cells) in culture. The complex caused inhibition of cellular proliferation in both osteoblast-like cells in culture, but the cytotoxicity was stronger in the normal (MC3T3E1) than in the tumoral (UMR106) osteoblasts. The effect of the complex in cell differentiation was tested through the specific activity of alkaline phosphatase of the UMR106 cells because they expressed a high activity of this enzyme. What occurs with other vanadium compounds [VO(oda)(H2O)2] is an inhibitory agent of osteoblast differentiation.

Vanadium and bone development: putative signaling pathways.

Vanadium is a trace element present in practically all cells in plants and animals. It exerts interesting actions in living systems. At pharmacological doses, vanadium compounds display relevant biological actions such as mimicking insulin and growth factors as well as having osteogenic activity. Some vanadium compounds also show antitumoral properties. The importance of vanadium in bone arises from the studies developed to establish the essentiality of this element in animals and humans. Bone tissue, where the element seems to play an important role, accumulates great amounts of vanadium. This paper reviews the physiology of osteoblasts, the involvement of different growth factors on bone development, and the effects of vanadium derivatives on the skeletal system of animal models and bone-related cells. Two cellular lines are discussed in particular; one derived from a rat osteosarcoma (UMR106) and the other is a nontransformed osteoblast cell line (MC3T3-E1). The effects of different growth factors and their mechanisms of action in these cellular lines are reviewed. These models of osteoblasts are especially useful in understanding the intracellular signaling pathways of vanadium derivatives in hard tissues. Vanadium uses an intricate interplay of intracellular mechanisms to exert different biochemical and pharmacological actions. The effects of vanadium derivatives on some cellular signaling pathways related to insulin are compiled in this review. The comprehension of these intracellular signaling pathways may facilitate the design of vanadium compounds with promising therapeutic applications as well as the understanding of secondary side effects derived from the use of vanadium as a therapeutic agent.

Three new vanadyl(IV) complexes with non-steroidal anti-inflammatory drugs (Ibuprofen, Naproxen and Tolmetin). Bioactivity on osteoblast-like cells in culture.

The synthesis and spectral and magnetic characterization of VO(2+) complexes with Ibuprofen (2-(4-isobutylphenyl)propionic acid), Naproxen (6-methoxy-alpha-methyl-2-naphthalene acetic acid) and Tolmetin (1-methyl-5-(4-methylbenzoyl)-1H-pyrrole-2-acetic acid) were studied. The complexes [VO(Ibu)(2)] x 5CH(3)OH, [VO(Nap)(2)] x 5CH(3)OH and [VO(Tol)(2)] were obtained from methanolic solutions under nitrogen atmosphere. The biological activities of these complexes on the proliferation of two osteoblast-like cells in culture (MC3T3E1 and UMR106) were compared with that of the vanadyl(IV) cation. The complexes exhibited different effects depending on the concentration and the cellular type, while no effect was observed for their parent drugs.

[Evaluation of the organisation of care for patients without appointment]. / Evaluación de la organización en la atención a pacientes sin cita.

OBJECTIVES: To calculate the improvement achieved in organising care for patients without appointments after introducing corrective measures. Second, to study the reasons for consultation, the duration of the symptoms because of which consultations without appointment take place, and the assessment by health personnel of the relevance of the emergency. DESIGN: Before-and-after study.Setting. Health Area 9, urban health centre, Leganés, Madrid. PATIENTS: All the patients attending consecutively without appointment in the study periods (assessment: November 1998; reassessment: November-December 1999). MAIN RESULTS: 588 patients were included in the assessment and 562 in the reassessment. They belonged to general practice and paediatrics. 5 quality criteria were set: recording of basic details; recording of reason for consultation; assessment in under ten minutes; attempt at prior appointment; and record in the clinical notes. As there were no references in the bibliography, they were agreed by the team. After corrective measures were taken, all criteria except the fourth improved significantly. Nonetheless, the previously set norm was only achieved in the second. The professionals thought 27% of the consultations without appointment in paediatrics were urgent, and 30% in general medicine. Mean duration of symptoms in non-urgent pathologies was 1.5 days in paediatrics and 13.4 in general medicine. The most common reasons for consultation were prescriptions (20%) in general medicine and digestive pathology (20.4%) in paediatrics. CONCLUSIONS: Setting up an improvement cycle at our health centre managed to raise the quality of care given to patients without appointment, although there are still some facets that could be improved so as to reach, at least, the standards set.

Evaluación de la organización en la atención a pacientes sin cita / Evaluation of the organisation of care for patients without appointment

Objetivo. Estimar la mejora alcanzada en la organización de la atención a pacientes sin cita tras la instauración de medidas correctoras. Secundariamente nos propusimos estudiar los motivos de consulta, el tiempo de duración de los síntomas por los que se consulta sin cita y la valoración por personal sanitario sobre la pertinencia de la urgencia. Diseño. Estudio antes-después. Emplazamiento. Área de Salud 9, centro de salud urbano. Leganés, Madrid. Pacientes. Todos los pacientes que de forma consecutiva acudieron sin cita en los períodos de estudio (evaluación: noviembre de 1998; reevaluación: noviembre-diciembre de 1999).Resultados principales. Se incluyeron 588 pacientes en la evaluación y 562 en la reevaluación, pertenecientes a medicina general y pediatría. Se establecieron 5 criterios de calidad relacionados con: registro de datos de filiación; registro del motivo de consulta; valoración en menos de 10 minutos; intento de cita previa y registro en la historia clínica. Ante la ausencia de referencias en la bibliografía, fueron consensuados por el equipo. Tras la instauración de medidas correctoras, todos mejoraron de forma significativa excepto el cuarto. No obstante, sólo se alcanzó el estándar previamente fijado en el segundo. Los profesionales consideraron urgentes un 27 por ciento de las consultas sin cita en pediatría y el 30 por ciento en medicina general. La duración media de los síntomas en las patologías no urgentes fue 1,5 días en pediatría y 13,4 en medicina general. Los motivos de consulta más frecuentes fueron recetas (20 por ciento) en medicina general y patología digestiva (20,4 por ciento) en pediatría. Conclusiones. La puesta en marcha de un ciclo de mejora en nuestro centro de salud ha conseguido elevar la calidad de la atención que se presta a los pacientes sin cita, si bien aún quedan algunos aspectos susceptibles de mejorar hasta alcanzar, al menos, los estándares establecidos. (AU)

Non-enzymatic glycosylation of a type I collagen matrix: effects on osteoblastic development and oxidative stress.

BACKGROUND: The tissue accumulation of protein-bound advanced glycation endproducts (AGE) may be involved in the etiology of diabetic chronic complications, including osteopenia. The aim of this study was to investigate the effect of an AGE-modified type I collagen substratum on the adhesion, spreading, proliferation and differentiation of rat osteosarcoma UMR106 and mouse non-transformed MC3T3E1 osteoblastic cells. We also studied the role of reactive oxygen species (ROS) and nitric oxide synthase (NOS) expression on these AGE-collagen mediated effects. RESULTS: AGE-collagen decreased the adhesion of UMR106 cells, but had no effect on the attachment of MC3T3E1 cells. In the UMR106 cell line, AGE-collagen also inhibited cellular proliferation, spreading and alkaline phosphatase (ALP) activity. In preosteoblastic MC3T3E1 cells (24-hour culture), proliferation and spreading were significantly increased by AGE-collagen. After one week of culture (differentiated MC3T3E1 osteoblasts) AGE-collagen inhibited ALP activity, but had no effect on cell number. In mineralizing MC3T3E1 cells (3-week culture) AGE-collagen induced a decrease in the number of surviving cells and of extracellular nodules of mineralization, without modifying their ALP activity. Intracellular ROS production, measured after a 48-hour culture, was decreased by AGE-collagen in MC3T3E1 cells, but was increased by AGE-collagen in UMR106 cells. After a 24-hour culture, AGE-collagen increased the expression of endothelial and inducible NOS, in both osteoblastic cell lines. CONCLUSIONS: These results suggest that the accumulation of AGE on bone extracellular matrix could regulate the proliferation and differentiation of osteoblastic cells. These effects appear to depend on the stage of osteoblastic development, and possibly involve the modulation of NOS expression and intracellular ROS pathways.

On the interaction of the vanadyl(IV) cation with lactose: inhibition effects of vanadyl(IV)/monosaccharide and disaccharide complexes upon alkaline phosphatase activity.

A new vanadyl(IV) complex of the disaccharide lactose was obtained in aqueous solution at pH = 13. The sodium salt of the complex, of composition Na4[VO(lactose)2].3H2O, has been characterized by elemental analysis and by ultraviolet-visible, diffuse reflectance, and infrared spectroscopies. Its magnetic susceptibility and thermal behavior were also investigated. The inhibitory effect on alkaline phosphatase activity was tested for this compound as well as for the vanadyl(IV) complexes with maltose, sucrose, glucose, fructose, and galactose. For comparative purposes, the free ligands and the vanadyl(IV) cation were also studied. The free sugars and the sucrose/VO complex exhibited the lowest inhibitory effect. Lactose-VO, maltose-VO, and the free VO2+ cation showed an intermediate inhibition potential, whereas the monosaccharide/VO complexes appeared as the most potent inhibitory agents.