Impact of the application of monosilicic acid to grapevine (Vitis vinifera L.) on the chemical composition of young red Mencia wines.

Impact of applying monosilicic acid to grapevines during ripening on chemical composition of Mencía red wines in an area where fungal diseases during summer are common was examined. The foliar application of monosilicic acid to grapevines led to a less oxidized wine, with lower levels of acetic acid, acetaldehyde, ethyl acetate and diacethyl; this should be considered as positive from a sensory point of view. Wines made with silicon-treated grapes also contained lower levels of gluconic acid and glycerol, which are chemical markers of wines made with botrytized grapes, as well as higher contents of total phenols, anthocyanins and tannins. Furthermore, the contents of several mid-chain alcohols were higher (p < 0.05) in wines made with grapes from silicon-treated plants.

Dynamic interactions between apex predators reveal contrasting seasonal attraction patterns.

Apex predators play important roles in ecosystem functioning and, where they coexist, intraguild interactions can have profound effects on trophic relationships. Interactions between predators range from intraguild predation and competition to facilitation through scavenging opportunities. Despite the increased availability of fine-scale GPS data, the determinants and outcomes of encounters between apex predators remain understudied. We used simultaneous GPS data from collared spotted hyaenas (Crocuta crocuta) and African lions (Panthera leo) in Hwange National Park, Zimbabwe, to determine the environmental conditions of the encounters between the two species, which species provoked the encounter, and which species dominated the encounter. Our results show that encounters between hyaenas and lions are mostly resource-related (over a carcass or around waterholes). In the wet season, encounters mainly occur at a carcass, with lions being dominant over its access. In the dry season, encounters mainly occur in the absence of a carcass and near waterholes. Movements of hyaenas and lions before, during, and after these dry-season encounters suggest two interference scenarios: a passive interference scenario whereby both predators would be attracted to waterholes but lions would leave a waterhole used by hyaenas because of prey disturbance, and an active interference scenario whereby hyaenas would actively chase lions from waterhole areas, which are prime hunting grounds. This study highlights the seasonal dynamics of predator interactions and illustrates how the relative importance of negative interactions (interference competition during the dry season) and positive interactions (scavenging opportunities during the wet season) shifts over the course of the year.

Características epidemiológicas de los pacientes con enfermedad crítica crónica / Epidemiological characteristics of patients with chronic critical illness

Resumen ANTECEDENTES: la población de pacientes que requiere cuidados intensivos por tiempo prologado se ha incrementado en las últimas décadas dando lugar a una nueva población de pacientes con enfermedad crítica crónica. OBJETIVO: describir las características epidemiológicas de los pacientes con enfermedad crítica crónica atendidos en el Hospital Regional de Alta Especialidad de Oaxaca. MATERIAL Y MÉTODO: estudio ambispectivo, observacional y descriptivo que incluyó los pacientes ingresados entre el 1 de enero de 2012 al 31 de diciembre de 2015. Se definió a los pacientes con enfermedad crítica crónica como aquéllos con ventilación mecánica prolongada definida como más de 21 días de ventilación mecánica por más de 6 horas al día. Los pacientes fueron seguidos hasta su egreso hospitalario o defunción. RESULTADOS: se incluyeron 284 pacientes, la incidencia de enfermedad crítica crónica fue de 10%. En los pacientes con enfermedad crítica crónica la escala de APACHE II fue de 19.4±9.7 y sin enfermedad crítica crónica fue de 15.94±8.6 (p=0.044), mientras que la escala SOFA en los pacientes con y sin enfermedad crítica crónica fue de 8.7±2.6 y 7.01±4.4, respectivamente (p=0.007). La media de días de estancia en la unidad de cuidados intensivos (UCI) en los pacientes con y sin enfermedad crítica crónica fue de 17.1±9.2 y 8±4.8, respectivamente (p=0.0000). Los días de estancia hospitalaria fueron 45.9±19.7 y 18.3±12.4 en los pacientes con y sin enfermedad crítica crónica, respectivamente (p=0.0000). La mortalidad en la UCI fue de 18 y 28% en los pacientes con y sin enfermedad crítica crónica, respectivamente (p=0.0000). La mortalidad hospitalaria fue de 50 y 16% en los pacientes con y sin enfermedad crítica crónica, respectivamente (p=0.0000). Egresaron del hospital 32 y 56% de los pacientes con y sin enfermedad crítica crónica, respectivamente (p=0.0000). CONCLUSIONES: la incidencia de enfermedad crítica crónica en nuestro estudio fue similar a la reportada en la bibliografía. La severidad de la enfermedad aguda al ingreso de acuerdo con las escalas SOFA y APACHE se relaciona con la aparición de enfermedad crítica crónica. Los pacientes con enfermedad crítica crónica tuvieron mayor estancia en la UCI, mayores días de sedación, así como mayor estancia y mortalidad hospitalarias.

Abstract BACKGROUND: Population of patients requiring intensive care for prolonged time has increased in recent decades resulting in a new population of patients with chronic critical illness (CCI). OBJECTIVE: To describe the epidemiologic characteristics of patients with chronic critical illness assisted at Regional Hospital of High Specialty of Oaxaca, Mexico. MATERIAL AND METHOD: A retrospective, prospective, observa tional and descriptive study included patients admitted from January 1st, 2012 to December 31st, 2015. Patients with CCI were defined as those with prolonged mechanical ventilation defined as ≥21 days of ventilation for ≥6 hours/day. Patients were followed until hospital discharge or death. RESULTS: 284 patients were included; the incidence of CCI was 10%. In patients with CCI the APACHE II score was 19.4±9.7 and without CCI was 15.94±8.6 (p=0.044), while the SOFA scale in patients with CCI was 8.7±2.6 and without CCI 7.01±4.4 (p=0.007). Days of ICU stay were 17.1±9.2 for patients with CCI and 8±4.8 without CCI (p=0.0000). Days of hospital stay were 45.9±19.7 in patients with CCI and 18.3±12.4 in patients without CCI (p=0.0000). ICU mortality was 18% in patients with CCI and 28% in patients without CCI (p=0.0000). The hospital mortality was 50% in patients with CCI and 16% in patients without CCI (p=0.0000). Discharged from hospital were 32% of patients with CCI and 56% of patients without CCI (p=0.0000). CONCLUSIONS: The incidence of CCI in our study was similar to that reported in the literature. Severity of acute disease income according to APACHE and SOFA scales was related to the development of CCI. Patients with chronic critical illness had higher ICU stay, more days sedation, longer hospital stays and increased hospital mortality.

Obtaining from grape pomace an enzymatic extract with anti-inflammatory properties.

Grape pomace, a winemaking industry by-product, is a rich source of bioactive dietary compounds. Using proteases we have developed an enzymatic process for obtaining a water-soluble extract (GP-EE) that contains biomolecules such as peptides, carbohydrates, lipids and polyphenols in soluble form. Of especial interest is its high polyphenol content (12%), of which 77% are flavonoids and 33% are phenolic acids. The present study evaluates in vitro the potential anti-inflammatory effect of GP-EE by monitoring the expression of inflammatory molecules on N13 microglia cells stimulated with lipopolysaccharide (LPS). GP-EE decreases the mRNA levels of the inflammatory molecules studied. The molecules under study were as follows: inducible nitric oxide synthase (iNOS), tumor necrosis factor- α (TNF-α), interleukin-1ß (IL-1ß), the ionized calcium binding adaptor molecule-1(Iba-1) and the Toll like receptor-4 (TLR-4), as well as the iNOS protein level in LPS-stimulated microglia. Our findings suggest that, as a result of its ability to regulate excessive microglial activation, GP-EE possesses anti-inflammatory properties. Therefore, acting as a chemopreventive agent, it may be of therapeutic interest in neurodegenerative diseases involving neuroinflammation. We can, therefore, propose GP-EE as a useful natural extract and one that would be beneficial to apply in the field of functional foods.

Antiproliferative and immunoactivatory ability of an enzymatic extract from rice bran.

The validation of natural products as source of functional foods or nutraceuticals has become an important issue in current health research. Thus, the present work has tested on MOLT-4 cells (human T cell acute lymphoblastic leukemic) the antiproliferative effect of a water-soluble enzymatic extract from rice bran (EERB). Present work shows that EERB induces cellular death in MOLT-4 cells in a dose-dependent way (0-10mg/mL) but not in non-tumoral lymphocytes. Flow cytometric analysis of MOLT-4 cells treated with EERB showed the presence of death cells by apoptosis rather than necrosis. Additionally, EERB also exerts an immunoactivatory effect on N13 microglia cells, by inducing TNF-alpha (tumour necrosis factor-α) expression, which plays a key role in the innate immune response to infection. Accordingly, we can propose EERB as a useful natural standardized extract with antiproliferative and immunoactivatory ability that would be beneficial to apply in the functional food field.

Protection against free radicals (UVB irradiation) of a water-soluble enzymatic extract from rice bran. Study using human keratinocyte monolayer and reconstructed human epidermis.

The antioxidant capacity of a water-soluble enzymatic extract from rice bran (EERB) has been tested in two cell models: keratinocyte monolayers and human reconstructed epidermis. Cells were incubated in culture medium in presence of different amounts of EERB and were UVB irradiated. Cell population assessment (MTT assay) and MDA (malonaldehyde) production were evaluated. The EERB did not induce cytotoxic effect for concentrations inferior or equal to 100 microg/mL. Human keratinocyte monolayers were protected of irradiation preventing 33% the lipid peroxidation process at concentration of 10 microg/ml of EEBR. In reconstructed human epidermis, 100 microg/mL decreased lipid peroxidation process by 44% (p<0.01) with regards to irradiated negative control. This effect was comparable to that of vitamin E at 600 microg/mL. Our data indicate that EERB is potentially able to efficiently counteract UVB-induced response. The EERB, diluted at 10% with water has very good skin compatibility. This product showed a sun protection factor of 4.8+/-0.3. Thus we can propose EERB as a useful natural standardized extract in skin photoprotection with promising applications in the field of dermatology.

Searching for DNA in museum specimens: a comparison of sources in a mammal species.

The number of genetic studies that use preserved specimens as sources of DNA has been steadily increasing during the last few years. Therefore, selecting the sources that are more likely to provide a suitable amount of DNA of enough quality to be amplified and at the minimum cost to the original specimen is an important step for future research. We have compared different types of tissue (hides vs. bones) from museum specimens of Iberian lynx and multiple alternative sources within each type (skin, footpad, footpad powder, claw, diaphysis, maxilloturbinal bone, mastoid process and canine) for DNA yield and probability of amplification of both mitochondrial and nuclear targets. Our results show that bone samples yield more and better DNA than hides, particularly from sources from skull, such as mastoid process and canines. However, claws offer an amplification success as high as bone sources, which makes them the preferred DNA source when no skeletal pieces have been preserved. Most importantly, these recommended sources can be sampled incurring minimal damage to the specimens while amplifying at a high success rate for both mitochondrial and microsatellite markers.

Dynamics of Haemophilus parasuis genotypes in a farm recovered from an outbreak of Glässer's disease.

Haemophilus parasuis is a colonizer of the upper respiratory tract of pigs, although it is better known as the etiological agent of Glässer's disease. Interestingly, several strains can be isolated from a single farm, as determined by both genotyping and serotyping. However, it is not known how an outbreak and the subsequent treatment affect the population of H. parasuis strains. In this study, a farm was studied during an outbreak of Glässer's disease and 1 year after antimicrobial treatment and elimination of clinical signs. Bacterial isolation was attempted from nasal swabs and lesions. After isolation, antimicrobial susceptibility, serotype and genotype were determined. Two different genotyping techniques, enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus sequence typing (MLST) were used. The H. parasuis strain that was isolated from lesions during the disease outbreak clustered with other virulent strains by both MLST and serotyping analysis. Nasal isolates were included in the corresponding nasal cluster by MLST, but they presented high variability by serotyping. These nasal isolates included serotypes previously classified as virulent and non-virulent. Finally, we found that during the antimicrobial treatment the diversity of strains isolated in the farm was affected and just one strain, which was resistant to the treatment, was detected. One year after the treatment strain diversity was back to normal (three strains).

Antibodies to Mycobacterium bovis in wild carnivores from Doñana National Park (Spain).

We conducted a retrospective serologic survey for antibodies against the MPB70 protein of Mycobacterium bovis in wild carnivores from Doñana National Park (southwestern Spain). Serum samples from 118 red foxes (Vulpes vulpes), 39 Iberian lynx (Lynx pardinus), 31 Eurasian badgers (Meles meles), five Egyptian mongoose (Herpestes ichneumon), four European genet (Genetta genetta), and one Eurasian otter (Lutra lutra) were analyzed using an indirect competitive enzyme-linked immunoassay. Antibodies against the MPB70 protein of M. bovis were detected in seven badgers, five foxes, and one lynx. The frequency of positive animals was significantly higher in badger (23%) than in lynx (3%) and fox (4%). Antibodies were not detected in other species. Annual antibody frequency peaked at 38% in badgers and 11% for red fox. These species may contribute to persistence of bovine tuberculosis in Doñana.

Role of p38 and inducible nitric oxide synthase in the in vivo dopaminergic cells' degeneration induced by inflammatory processes after lipopolysaccharide injection.

Accumulating evidences suggest that neuroinflammation is involved in the progressive death of dopaminergic neurons in Parkinson's disease. Several studies have shown that intranigral injection of lipopolysaccharide induces inflammation in the substantia nigra leading to death of tyrosine hydroxylase-positive cells. To better understand how the inflammatory response gives rise to neurotoxicity we induced inflammation in substantia nigra by injecting lipopolysaccharide. The damage of substantia nigra dopaminergic neurons was evaluated by immunohistochemistry, reverse transcription-PCR and Western blot analysis of tyrosine hydroxylase. In parallel, activation of microglial cells, a hallmark of inflammation in CNS, was revealed by immunohistochemistry. Similarly the expression of molecules involved in the inflammatory response and apoptotic pathway was also tested, such as cytokines (tumor necrosis factor-alpha, interleukin-1beta, interleukin-6), inducible nitric oxide synthase and caspase-11. Tyrosine hydroxylase expression (both mRNA and protein) started to decrease around 3 days post-injection. At the mRNA level, our results showed that the cytokines expression peaked shortly (3-6 h) after lipopolysaccharide injection, followed by the induction of inducible nitric oxide synthase and caspase-11 (14 h). However, inducible nitric oxide synthase protein peaked at 24 h and lasted for 14 days. The lipopolysaccharide-induced loss of substantia nigra dopaminergic neurons was partially inhibited by co-injection of lipopolysaccharide with S-methylisothiourea, an inducible nitric oxide synthase inhibitor. Co-injections of lipopolysaccharide with SB203580, a p38 MAP kinase inhibitor, reduced inducible nitric oxide synthase and caspase-11 mRNA expression, and also rescued dopaminergic neurons in substantia nigra. In summary, this is the first report to describe in vivo the temporal profile of the expression of these inflammatory mediators and proteins involved in dopaminergic neuronal death after intranigral injection of lipopolysaccharide. Moreover data strongly support that lipopolysaccharide-induced dopaminergic cellular death in substantia nigra could be mediated, at least in part, by the p38 signal pathway leading to activation of inducible nitric oxide synthase and caspase-11.

Bovine tuberculosis in a free ranging red fox (Vulpes vulpes) from Doñana National Park (Spain).

During 1997 and 1998, a survey of Iberian carnivores was conducted to study the epidemiology of bovine tuberculosis in the Doñana National Park and surrounding areas in southwestern Spain. Post-mortem examinations were done on seven red foxes (Vulpes vulpes), two Egyptian mongoose (Herpestes ichneumon), one weasel (Mustela nivalis), two genets (Genetta genetta), one Iberian lynx (Lynx pardinus), one Eurasian badger (Meles meles), and two polecats (Mustela putorius). Lesions suggestive of bovine tuberculosis were not detected but, in culture, Mycobacterium bovis was isolated from the retropharyngeal lymph nodes of one adult male red fox. This is the first report of M. bovis infection in red fox in Spain.

Phylogenetic and phylogeographic analysis of Iberian lynx populations.

The Iberian lynx (Lynx pardinus), one of the world's most endangered cat species, is vulnerable due to habitat loss, increased fragmentation of populations, and precipitous demographic reductions. An understanding of Iberian lynx evolutionary history is necessary to develop rational management plans for the species. Our objectives were to assess Iberian lynx genetic diversity at three evolutionary timescales. First we analyzed mitochondrial DNA (mtDNA) sequence variation to position the Iberian lynx relative to other species of the genus LYNX: We then assessed the pattern of mtDNA variation of isolated populations across the Iberian Peninsula. Finally we estimated levels of gene flow between two of the most important remaining lynx populations (Doñana National Park and the Sierra Morena Mountains) and characterized the extent of microsatellite locus variation in these populations. Phylogenetic analyses of 1613 bp of mtDNA sequence variation supports the hypothesis that the Iberian lynx, Eurasian lynx, and Canadian lynx diverged within a short time period around 1.53-1.68 million years ago, and that the Iberian lynx and Eurasian lynx are sister taxa. Relative to most other felid species, genetic variation in mtDNA genes and nuclear microsatellites were reduced in Iberian lynx, suggesting that they experienced a fairly severe demographic bottleneck. In addition, the effects of more recent reductions in gene flow and population size are being manifested in local patterns of molecular genetic variation. These data, combined with recent studies modeling the viability of Iberian lynx populations, should provide greater urgency for the development and implementation of rational in situ and ex situ conservation plans.

Value of high-performance liquid chromatographic analysis of anthocyanins in the differentiation of red grape cultivars and red wines made from them.

A HPLC method that allows the separation of several anthocyanins present in red grapes and red wines, using a linear gradient of acetonitrile in water at pH 1.3, using perchloric acid as an acid modifier, is described. Data clearly show that the anthocyanins profile of red grapes may be complex, but quite different for each cultivar studied. Thus, those molecules may be used as chemotaxonomic markers for classifying red grape cultivars. However, the anthocyanin profile of red wines clearly differs from that presented by grapes employed in making it, because red wine contains a higher relative amount of malvidin-3-O-glucoside than grapes, and the relative amount of other anthocyanins in wines is usually lower than in grapes. Therefore, the use of anthocyanins present in wines to determine the grape cultivar used for winemaking needs a careful evaluation of the influence of different technological procedures on the anthocyanins fingerprint.

Analysis of several phenolic compounds with potential antioxidant properties in grape extracts and wines by high-performance liquid chromatography-photodiode array detection without sample preparation.

A RP-HPLC method that allows the separation of several types of phenolic compounds present in grapes and wines by direct injection of samples, using a binary gradient with solvents free of salts and photodiode array detection is described. Results show that more than 15 different phenolic molecules with antioxidant properties (flavan-3-ols, anthocyanins, cinnamic acid derivatives, flavonol derivatives and trans-resveratrol) may be separated in a single run by direct injection of red wine. The method is also valuable for the analysis of these compounds in white wine and in skins, seeds and pulp extracts of red and white grapes.

GABAA and alpha-amino-3-hydroxy-5-methylsoxazole-4-propionate receptors are differentially affected by aging in the rat hippocampus.

We have investigated the age-dependent modifications in the expression of eight different subunits of the gamma-aminobutyric acid, type A (GABA(A)) receptor (alpha1, alpha2, alpha3, alpha5, beta2, beta3, gamma2S, and gamma2L) and all four subunits of the alpha-amino-3-hydroxy-5-methylsoxazole-4-propionate (AMPA) receptor (GluR1-4) in the hippocampus of 24-month-old rats. All aged hippocampi displayed a remarkable increase (aged/adult ratio, 3.53 +/- 0.54) in the mRNA levels of the short version of the gamma2 subunit in parallel with a similar increase in the gamma2 subunit protein (aged/adult ratio, 2.90 +/- 0.62). However, this increase was not observed in the mature receptor. On the other hand, the expression of the different alpha subunit mRNAs increased moderately with aging, displaying a heterogeneous pattern. The most frequent modification consisted in an increase in the expression of the alpha1 subunit mRNA (aged/adult ratio, 1.26 +/- 0.18), in parallel with a similar increase on the alpha1 protein (aged/adult ratio, 1. 27 +/- 0.12) and in the alpha1 incorporated to the assembled GABA(A) receptor (tested by immunoprecipitation; aged/adult ratio, = 1.20 +/- 0.10). However, in the same hippocampal samples, no major modifications were observed on the expression of the AMPA receptor subunits. As a whole, these results indicated the existence of an increased expression of the GABA(A) receptor subunits and a preservation of the AMPA receptor at the hippocampal formation. These modifications could reflect the existence of specific deficiencies (neuronal loss and/or deafferentiation) on the GABAergic system in the aged rats.

[Chronobiological profile of arterial blood pressure and heart rate in a family group determined by automatic monitoring]. / El perfil cronobiológico de tensión arterial y de frecuencia cardiaca en un grupo familiar, determinado mediante monitorizacion automática.

Over a week, blood pressure (systolic, mean and diastolic) and heart rate were determined in a family, by means of automatic, uninterrupted monitoring. The chronobiological profile for each family member was prepared in time series of various periodicities. An ample circadian component and a lesser circaseptan component were apparent. Clear phase differences were identified among the four cardiovascular variables studied. The chronobiological profile of the children was closer to that of the father than to that of the mother.