Progress and challenges in valorisation of biomass waste from ornamental trees pruning through pyrolysis processes. Prospects in the bioenergy sector.

Nowadays, the scarcity of energy resources is promoting the search for alternative energy sources, boosting interest in the use of forest lignocellulosic residue in the energy sector. In this study, the focus is on the energy recovery from two lignocellulosic residues originated during the pruning of ornamental trees (Horse Chestnut, CI, and False Acacia, FA). Both conventional and flash pyrolysis techniques were applied. The experimental pyrolysis variables were obtained from the study of the thermal behaviour of the pruning residues in thermogravimetric analysis. It was carried out under 5 heating rates and kinetic parameters were estimated using Flynn-Wall-Ozawa method. Results denoted higher maximum mass loss rate values for the same release temperature regions under FA experiments. Also, FA samples had lower final residues for the processes. However, activation energy values were so close for both species. FA was also linked to the faster reactions according frequency factor outcomes. Conventional pyrolysis of pruning residues was carried out in a horizontal oven of original design at a heating rate of 25 °C/min, at 750 °C and 60 min of permanence at that temperature; flash pyrolysis was tested in that oven at 750 and 850 °C. In these pyrolysis processes, three fractions were obtained: bio-char, bio-oil and gas. The physicochemical attributes of the bio-chars suggested their potential utility as biofuels (28.4-29.8 MJ/kg), adsorbent precursors or soil additives. Conventional pyrolysis bio-oils had a dominant monoaromatic hydrocarbons nature, with phenols being the most abundant (≥60%), while flash bio-oils contain mainly polycyclic aromatic hydrocarbons. Conventional pyrolysis gases contained up to 60 vol% of CO2; flash pyrolysis gases had high combustible gas content (CO, CH4, H2) and a low CO2 content (<25 vol%). As a result, their calorific value (18.06 MJ/kg) exhibited a threefold increase compared to the gas produced through conventional pyrolysis (6.04 MJ/kg).

Indications and results of sentinel lymph node biopsy in breast cancer: Changes observed in 2018 compared to 2012.

BACKGROUND AND OBJECTIVE: Over the last 15 years, breast cancer (BC) treatment has undergone numerous changes, which have also affected the indications for Sentinel Lymph Node Biopsy (SLNB) as well as the procedures depending on its outcome. The aim of this study is to compare the results of the SLNB carried out at our Center during 2012, when we stopped performing an axillary lymph node dissection (ALND) after the finding of a micrometastasis, with those conducted in 2018, when we started applying Z011 criteria. MATERIALS AND METHODS: We have performed a comparative retrospective observational study, including cN0 BC patients that underwent a SLNB in 2012 versus those that underwent this procedure in 2018. RESULTS: 174 patients from 2012 and 165 from 2018 were studied. We found significant differences between the two groups: in 2018 there were fewer lobular invasive cancers (14 vs 28) (P < .05), a smaller mean pathological size (P < .001), a higher proportion of HER2 and triple negative tumors (28 vs 49) (P < .01) and, finally, an increase in use of neoadjuvant treatments (42.0% vs 6.7%). Regarding axillary involvement, we observed a decrease in both the presence of a positive sentinel node (24.1% in 2018 vs 42.4% in 2012) (P < .0001) as well as in the proportion of ALND performed (12.6% in 2018 vs 21.2% in 2012) and the presence of positive non sentinel nodes after a ALND (59.1% vs74.3%) (ns) CONCLUSION: Although the 2018 cohort that underwent SLNB had more aggressive tumors, there were fewer positive SN and ALND performed. This is probably due to a more accurate radiological diagnosis with ultrasound, that enables to detect cN1 cases before surgery, and to the increasing use of neoadjuvant treatments that may downstage the axilla.

Relación del insomnio con la calidad de vida en una población de adultos mayores / Relationship of insomnia with quality of life in a population of older adults

Antecedentes: el insomnio es considerado un problema de salud pública por su alta prevalencia, se manifiesta por una percepción subjetiva de insatisfacción por la cantidad del sueño, dificultad para iniciar o mantener el sueño, despertar antes de lo deseado, incapacidad para volver a dormir y que ocurre a pesar de contar con las condiciones adecuadas, presentándose con frecuencia en adultos mayores afectando la calidad de vida, además, altera la funcionalidad física, mental y social de las personas mayores , y es un factor predisponente para deterioro cognitivo y depresión. Objetivo: determinar la relación del insomnio con la calidad de vida en una población de adultos mayores. Metodología: estudio analítico, transversal y comparativo, en una población de 107 adultos mayores de la Ciudad de México, con insomnio y sin insomnio, a los cuales se les aplicó la escala de Atenas para diagnosticar insomnio; como pruebas de tamizaje para deterioro cognitivo el MiniMental de Folstein, la Escala de Depresión Geriátrica Yesavage para probable depresión, y para calidad de vida el Cuestionario WHOQoL-BrefdelaOMS. Los datos fueron analizados a través de X2y t de Student y como estimador de riesgos razón de momios con un intervalo de confianza al 95%. Resultados:laprevalenciadeinsomniofuedel57%;enlascaracterísticassociodemográficas no se encontraron diferencias significativas; en las pruebas de tamizaje, se observó una relación estadísticamente significativa entre la presencia de insomnio y depresión (p<0.05); se encontró que los adultos mayores con insomnio tienen 6 veces mayor riesgo de presentar depresión en comparación con los que no lo presentan (RM 7.292.9, IC95%=2.319-22.925, p<0.05); respecto a la relación del insomnio con la calidad de vida, en los pacientes con insomnio fue estadísticamente significativa con un puntaje menor en la calidad de vida global(con insomnio 93 ± 13.7 vs. sin insomnio 102 ± 11.9, p<0.05); en la relación del insomnio y el nivel de calidad de vida, se encontró que los adultos mayores sin insomnio presentan una calidad de vida alta, en comparación con los adultos mayores con insomnio, donde se observó que es promedio (con Insomnio 70% vs. sin insomnio 30%, p<0.05); de acuerdo con los valores promedio de las diferentes dimensiones de la calidad de vida y su relación con el insomnio, se detectó que el grupo con insomnio obtuvo puntajes menores en las dimensiones salud física, aspectos psicológicos, relaciones sociales y medio ambiente observándose diferencias estadísticamente significativas entre los dos grupos; se detectó que el insomnio aumenta el riesgo de percibir la calidad de vida como promedio-mala, primordialmente en las dimensiones salud física, aspectos psicológicos y relaciones sociales. Conclusiones: La población de estudio con insomnio presentó alteración en las 4 dimensiones de la calidad de vida (salud física, aspectos psicológicos, relaciones sociales y medio ambiente).

Background: insomnia is considered as a public health problem because of its high prevalence, is manifested by a subjective perception of dissatisfaction with the amount of sleep, difficulty initiating or maintaining sleep, to wake up earlier than desired , inability to go back to sleep. These events happen despite having the suitable conditions, occurring frequently in older adults affecting the quality of life, also alters the physical, mental and social functionality of older people, and it's a predisposing factor for cognitive impairment and depression. Objective: to determine the relationship of insomnia with quality of life in a population of older adults. Methodology: analytical, cross-sectional and comparative study in a population of 107older adults in Mexico City, with insomnia and without insomnia, to which the Athenas scale was applied to diagnose insomnia; as screening tests for cognitive impairment the Folstein Mini Mental, the Yesavage Geriatric Depression Scale for probable depression, and the WHOQoL-Bref Questionnaire of WHO, for quality of life were used. The data were analyzed using X2and Student's t test,and as an estimator odds ratio risks with a 95%confidence interval. Results: the prevalence of insomnia was 57%; no significant differences were found in sociodemographic characteristics; in the screening tests, a statistically significant relationship was observed between the presence of insomnia and depression (p<0.05); it was found that older adults with insomnia have a 6times greater risk of presenting depression compared to those without it (OR 7,292.9, 95% CI = 2,319-22,925, p<0.05); with respect to the relationship between insomnia and quality of life, in patients with insomnia it was statistically significant with a lower score in global quality of life (with insomnia 93 ± 13.7vs. without insomnia 102 ± 11.9, p <0.05); in the relationship between insomnia and the level of quality of life, it was found that older adults without insomnia have a high quality of life, compared to older adults with insomnia, where it was observed that it is average (with insomnia 70%vs. without insomnia 30%, p<0.05); according to the average values of the different dimensions of quality of life and their relationship with insomnia, it was detected that the group with insomnia obtained lower scores in the dimensions of physical health, psychological aspects, social relations,and environment, observing statistically significant differences between the two groups. Insomnia was found to increase the risk of perceiving quality of life as average-poor, primarily in the dimensions of physical health, psychological aspects, and social relationships. Conclusions: The study population with insomnia presented alteration in the 4dimensions of quality of life (physical health, psychological aspects, social relationships,and environment).

Continuous aqueous phase reforming of a synthetic brewery wastewater with Pt/C and PtRe/C catalysts for biohydrogen production.

This work investigates H2 production through aqueous phase reforming (APR) of synthetic brewery wastewater in a continuous fixed bed reactor with Pt and PtRe (3 wt %) catalysts supported on activated carbon. The influence of weight hourly space velocity (WHSV) and superficial Ar gas flow velocity (VAr) was assessed for the sake of optimisation, while reaction temperature and pressure were maintained at 225 °C and 28 bar, respectively. H2 production was found to be higher using the PtRe catalyst at the lowest WHSV (0.03 h-1) and highest VAr (0.8 cm s-1). The comparison of the maximum H2 production obtained in this work (27.9 µmol min-1) with other treatment processes shows the potential of the application of APR process for H2 production from brewery wastewater. Despite the different reaction conditions tested, the catalysts showed deactivation with time on stream, which was related to the formation of solid deposits on the surface of the catalysts. Therefore, future research should be related to the development of more stable catalysts, strategies that avoid deactivation by coking and regeneration processes.

Economic impact of improving patient safety using Sugammadex for routine reversal of neuromuscular blockade in Spain.

BACKGROUND: Neuromuscular blocking (NMB) agents are often administered to facilitate tracheal intubation and prevent patient movement during surgical procedures requiring the use of general anesthetics. Incomplete reversal of NMB, can lead to residual NMB, which can increase the risk of post-operative pulmonary complications. Sugammadex is indicated to reverse neuromuscular blockade induced by rocuronium or vecuronium in adults. The aim of this study is to estimate the clinical and economic impact of introducing sugammadex to routine reversal of neuromuscular blockade (NMB) with rocuronium in Spain. METHODS: A decision analytic model was constructed reflecting a set of procedures using rocuronium that resulted in moderate or deep NMB at the end of the procedure. Two scenarios were considered for 537,931 procedures using NMB agents in Spain in 2015: a scenario without sugammadex versus a scenario with sugammadex. Comparators included neostigmine (plus glycopyrrolate) and no reversal agent. The total costs for the healthcare system were estimated from the net of costs of reversal agents and overall cost offsets via reduction in postoperative pneumonias and atelectasis for which incidence rates were based on a Spanish real-world evidence (RWE) study. The model time horizon was assumed to be one year. Costs were expressed in 2019 euros (€) and estimated from the perspective of a healthcare system. One-way sensitivity analysis was carried out by varying each parameter included in the model within a range of +/- 50%. RESULTS: The estimated budget impact of the introduction of sugammadex to the routine reversal of neuromuscular blockade in Spanish hospitals was a net saving of €57.1 million annually. An increase in drug acquisition costs was offset by savings in post-operative pulmonary events, including 4806 post-operative pneumonias and 13,996 cases of atelectasis. The total cost of complications avoided was €70.4 million. All parameters included in the model were tested in sensitivity analysis and were favorable to the scenario with sugammadex. CONCLUSIONS: This economic analysis shows that sugammadex can potentially lead to cost savings for the reversal of rocuronium-induced moderate or profound NMB compared to no reversal and reversal with neostigmine in the Spanish health care setting. The economic model was based on data obtained from Spain and from assumptions from clinical practice and may not be valid for other countries.

Palbociclib in combination with endocrine therapy versus capecitabine in hormonal receptor-positive, human epidermal growth factor 2-negative, aromatase inhibitor-resistant metastatic breast cancer: a phase III randomised controlled trial-PEARL.

BACKGROUND: Palbociclib plus endocrine therapy (ET) is the standard treatment of hormone receptor-positive and human epidermal growth factor receptor 2-negative, metastatic breast cancer (MBC). However, its efficacy has not been compared with that of chemotherapy in a phase III trial. PATIENTS AND METHODS: PEARL is a multicentre, phase III randomised study in which patients with aromatase inhibitor (AI)-resistant MBC were included in two consecutive cohorts. In cohort 1, patients were randomised 1 : 1 to palbociclib plus exemestane or capecitabine. On discovering new evidence about estrogen receptor-1 (ESR1) mutations inducing resistance to AIs, the trial was amended to include cohort 2, in which patients were randomised 1 : 1 between palbociclib plus fulvestrant and capecitabine. The stratification criteria were disease site, prior sensitivity to ET, prior chemotherapy for MBC, and country of origin. Co-primary endpoints were progression-free survival (PFS) in cohort 2 and in wild-type ESR1 patients (cohort 1 + cohort 2). ESR1 hotspot mutations were analysed in baseline circulating tumour DNA. RESULTS: From March 2014 to July 2018, 296 and 305 patients were included in cohort 1 and cohort 2, respectively. Palbociclib plus ET was not superior to capecitabine in both cohort 2 [median PFS: 7.5 versus 10.0 months; adjusted hazard ratio (aHR): 1.13; 95% confidence interval (CI): 0.85-1.50] and wild-type ESR1 patients (median PFS: 8.0 versus 10.6 months; aHR: 1.11; 95% CI: 0.87-1.41). The most frequent grade 3-4 toxicities with palbociclib plus exemestane, palbociclib plus fulvestrant and capecitabine, respectively, were neutropenia (57.4%, 55.7% and 5.5%), hand/foot syndrome (0%, 0% and 23.5%), and diarrhoea (1.3%, 1.3% and 7.6%). Palbociclib plus ET offered better quality of life (aHR for time to deterioration of global health status: 0.67; 95% CI: 0.53-0.85). CONCLUSIONS: There was no statistical superiority of palbociclib plus ET over capecitabine with respect to PFS in MBC patients resistant to AIs. Palbociclib plus ET showed a better safety profile and improved quality of life.

[Analysis of the predictive value of preventive isolation criteria in the intensive care unit]. / Análisis del valor predictivo de los criterios de aislamiento preventivo en una unidad de cuidados intensivos.

OBJECTIVE: To assess the diagnostic accuracy of the criteria used to detect patients carrying multiresistant microorganisms (MRMs). DESIGN: A prospective observational study was carried out from May 2014 to May 2015. SETTING: Polyvalent Intensive Care Unit. PATIENTS: A cohort of consecutively admitted patients meeting the following criteria for preventive isolation according to the "Zero Resistance" project: hospital length of stay>4 days in the last three months ("hospital"); antibiotherapy during one week in the last month ("antibiotic"); institutionalized patients or recurrent contact with healthcare ("institution or care"); MRM carrier in the last 6 months ("previous MRM"). VARIABLES: Demographic data, culture results and isolation time. A multivariate analysis was performed using multiple logistic regression between each of the risk factors and patient MRM carrier status. RESULTS: During the study period, 575 patients were admitted, of which 28% met the isolation criteria (162). Fifty-one (31%) were MRM carriers. Of the patients who did not meet the criteria, 29 (7%) were carriers. In the multivariate analysis, the only variable independently associated to carrier status was "previous MRM", with OR=12.14 (95%CI 4.24-34.77). CONCLUSIONS: The only criterion independently associated with the ability to detect patients with MRMs upon admission to the ICU was the existence of "previous MRM".

Aqueous phase reforming coupled to catalytic wet air oxidation for the removal and valorisation of phenolic compounds in wastewater.

Aqueous phase reforming (APR) coupled to catalytic wet air oxidation (CWAO) has been investigated as an approach to remove phenolic compounds from wastewater, converting them into valuable gases. Partial oxidation of phenol was achieved in the first CWAO stage trying to minimize mineralization so to allow a high yield to valuable gases in the second APR stage. APR runs were carried out with different mixtures of compounds corresponding to phenol oxidation pathway (phenol, quinones, long and short chain acids) and representing different degrees of oxidation in CWAO stage. A range of TOC and COD removal (74-90%) was observed in APR stage for the single compounds, with higher removal for long chain acids. Likewise, long chain acids provided with the highest conversion to gases. APR of mixtures rich in acids gave the highest yield to CH4 (11.0 mmol CH4/g TOCinitial). H2 production was low in all cases, due to competing direct conversion of long and short chain acids into CH4. TOC and COD removal from wastewater was similar in APR-CWAO and APR, however the conversion to gases and the yield to CH4 were markedly higher for APR-CWAO, thus overcoming the difficulties previously observed in the direct APR of phenol.

Combination of dietary glycaemic index and fasting time prior to slaughter as strategy to modify quality of pork.

This study evaluates the effect of the dietary glycaemic index (low: L-GI vs. high: H-GI), pre-slaughter fasting times (17 h: f17 vs. 19 h: f19 vs. 22 h: f22), and their interaction, on certain meat quality characteristics. Greater duration of fasting resulted in lesser drip loss of muscle, greater LIPE expression (lipolysis) and 10% greater TBARS, with PUFA being the main source of free-fatty acid production. L-GI diets produced a greater drip loss, lipolytic activity and lesser C18:1n-9 relative mobilization, suggesting this group had greater glycolytic potential than H-GI. An interaction was found in meat characteristics related to fat metabolism. The IRS2 gene expression was greater in pigs fed L-GI diets, but only in a low fasting period. LIPE gene expression was greater for L-GI-f17 group than for L-GI-f22. Lesser n-6 PUFA but greater MUFA were found in the free fatty acid fraction of L-GI- f17 group thus resulting in a faster lipolysis initiation in this group.

Human sperm decondensation in vitro is related to cleavage rate and embryo quality in IVF.

PURPOSE: To investigate whether the ability of human spermatozoa to decondense in vitro in the presence of heparin (Hep) and glutathione (GSH) is related to assisted reproduction (ART) success. METHODS: Cross-sectional pilot study involving male partners of 129 infertile couples undergoing ICSI with (45) or without (84) donor oocytes at two infertility clinics in CABA, Argentina, between October 2012 and December 2013. In vitro decondensation kinetics with Hep and GSH and DNA fragmentation (TUNEL) were determined on the same sample used for ICSI. The possible relationship of decondensation parameters (maximum decondensation and decondensation velocity) and TUNEL values with ART success was evaluated. RESULTS: Embryo quality correlated positively with decondensation velocity (D60/D30) (Spearman's correlation, p < 0.05). According to D60/D30 values, patients were classified as slow decondensers (SlowD) (n = 68) or fast decondensers (FastD) (n = 61). Embryo quality was better in FastD (unpaired t test, p < 0.05). FastD and SlowD were subdivided according to use of donor oocytes. Among SlowD, biochemical and clinical pregnancy rates per transfer were significantly higher in donor (n = 19) vs. in non-donor (n = 31) cycles (Fisher's exact test, p < 0.05). TUNEL values were not related to embryo quality, but no clinical pregnancies or live births were achieved in TUNEL+ SlowD (n = 7). CONCLUSION: Decondensation kinetics of human spermatozoa in vitro with Hep and GSH could be related to embryo quality and ART success.

Prognostic role for the derived neutrophil-to-lymphocyte ratio in early breast cancer: a GEICAM/9906 substudy.

PURPOSE: Elevated markers of host inflammation, a hallmark of cancer, have been associated with worse outcomes in several solid tumors. Here, we explore the prognostic role of the derived neutrophil-to-lymphocyte ratio (dNLR), across different tumor subtypes, in patients with early breast cancer. PATIENTS AND METHODS: This was a retrospective analysis of 1246 patients with lymph node-positive, operable early breast cancer enrolled in the GEICAM/9906 trial, a multicenter randomized phase 3 study evaluating adjuvant chemotherapy. dNLR was calculated as the ratio of neutrophils and the difference between total leukocytes and neutrophils in peripheral blood before chemotherapy. Disease-free survival (DFS) and overall survival were explored using a Cox proportional hazard analysis. RESULTS: The analysis comprised 1243 (99.8%) patients with dNLR data, with a median follow-up of 10 years. Data on intrinsic subtypes were available from 818 (66%) patients (luminal A 34%, luminal B 32%, HER2-enriched 21% and basal-like 9%). Median dNLR was 1.35 [interquartile range (IQR) 1.08-1.71]. In the whole population, dNLR was not prognostic after adjustment for clinico-pathological factors. However, dNLR ≥ 1.35 was independently associated with worse DFS in the hormone receptor-negative/HER2+ population (HR 2.86; p = 0.038) and in patients with one to three lymph node metastases (HR 1.32, p = 0.032). There was a non-significant association with worse DFS in non-luminal and in HER2-enriched tumors (HR 1.40, p = 0.085 and HR 1.53, p = 0.067). No significant interaction was observed between the treatment arm and dNLR. CONCLUSION: Elevated dNLR appears to be an adverse prognostic factor in hormone receptor-negative early breast cancer. TRIAL REGISTRATION: EudraCT: 2005-003108-12 (retrospectively registered 28/06/2005). ClinicalTrials.gov Identifier: NCT00129922 (retrospectively registered 10/08/2005). Results of this study were presented in part at the 2016 ESMO conference October 7-11, 2016, Copenhagen, Denmark (oral presentation).

Outcomes of single versus double hormone receptor-positive breast cancer. A GEICAM/9906 sub-study.

BACKGROUND: Retrospective data suggest better outcomes for patients with double hormonal receptor (oestrogen [ER] and progesterone receptor [PgR])-positive (dHR+) early breast cancer, compared with single hormonal receptor-positive, sHR+, (ER+/PgR- or ER-/PgR+) disease. Here, we evaluate the classification according to intrinsic subtypes and clinical outcomes of sHR+ versus dHR+ in HER2-negative breast cancer patients enrolled in GEICAM/9906 study (NCT00129922). METHODS: Archival tumours were retrieved retrospectively for the analysis of ER, PgR and HER2 status and classified into intrinsic subtypes using the PAM50 gene expression assay. Disease-free survival (DFS) and overall survival (OS) were explored using a Cox proportional hazard analysis. RESULTS: Data on intrinsic subtypes were available in 571 (50%) patients with ER+ and/or PR+, and HER2-negative primary tumours. The incidence of luminal A and luminal B subtypes were 52%/36% in dHR+ tumours (ER+/PgR+), and 15%/58% in ER+/PgR-tumours. ER-/PgR+ tumours were mainly luminal A (52%). Compared with ER+/PgR+ patients, DFS was similar in ER-/PgR+ (hazard ratio [HR] 1.15, 95% confidence interval [CI] 0.57-2.34, p = 0.70) but worse in ER+/PgR- patients (HR 1.60, 95% CI 1.12-2.28, p < 0.01). Similar results were observed for OS (HR 1.50, p = 0.30 and HR 1.86, p < 0.01, respectively). CONCLUSIONS: The ER+/PgR- group is characterised by higher proliferation and worse outcomes. In spite of the ER-/PgR+ subgroup resembles ER+/PgR+ disease in terms of molecular subtypes and outcomes, the small number of patients in this subgroup prevents from drawing any conclusions. TRIAL REGISTRATION: EudraCT: 2005-003108-12 (retrospectively registered 28/06/2005). CLINICALTRIALS. GOV IDENTIFIER: NCT00129922 (retrospectively registered 10/08/2005).

Secreted protein extract analyses present the plant pathogen Alternaria alternata as a suitable industrial enzyme toolbox.

Lignocellulosic plant biomass is the most abundant carbon source in the planet, which makes it a potential substrate for biorefinery. It consists of polysaccharides and other molecules with applications in pharmaceutical, food and feed, cosmetics, paper and textile industries. The exploitation of these resources requires the hydrolysis of the plant cell wall, which is a complex process. Aiming to discover novel fungal natural isolates with lignocellulolytic capacities, a screening for feruloyl esterase activity was performed in samples taken from different metal surfaces. An extracellular enzyme extract from the most promising candidate, the natural isolate Alternaria alternata PDA1, was analyzed. The feruloyl esterase activity of the enzyme extract was characterized, determining the pH and temperature optima (pH 5.0 and 55-60 °C, respectively), thermal stability and kinetic parameters, among others. Proteomic analyses derived from two-dimensional gels allowed the identification and classification of 97 protein spots from the extracellular proteome. Most of the identified proteins belonged to the carbohydrates metabolism group, particularly plant cell wall degradation. Enzymatic activities of the identified proteins (ß-glucosidase, cellobiohydrolase, endoglucanase, ß-xylosidase and xylanase) of the extract were also measured. These findings confirm A. alternata PDA1 as a promising lignocellulolytic enzyme producer. SIGNIFICANCE: Although plant biomass is an abundant material that can be potentially utilized by several industries, the effective hydrolysis of the recalcitrant plant cell wall is not a straightforward process. As this hydrolysis occurs in nature relying almost solely on microbial enzymatic systems, it is reasonable to infer that further studies on lignocellulolytic enzymes will discover new sustainable industrial solutions. The results included in this paper provide a promising fungal candidate for biotechnological processes to obtain added value from plant byproducts and analogous substrates. Moreover, the proteomic analysis of the secretome of a natural isolate of Alternaria sp. grown in the presence of one of the most used vegetal substrates on the biofuels industry (sugar beet pulp) sheds light on the extracellular enzymatic machinery of this fungal plant pathogen, and can be potentially applied to developing new industrial enzymatic tools. This work is, to our knowledge, the first to analyze in depth the secreted enzyme extract of the plant pathogen Alternaria when grown on a lignocellulosic substrate, identifying its proteins by means of MALDI-TOF/TOF mass spectrometry and characterizing its feruloyl esterase, cellulase and xylanolytic activities.

Reproductive responses to sexually active buck of does treated with melatonin when body weight/body condition is increasing or decreasing.

When the sexual activity of bucks is minimal, there is a minimal male effect on does regardless of their body weight (BW)/body condition (BC) and whether does are treated with melatonin or not. The study examines whether sexually active bucks can induce an adequate male effect in does with an increasing or decreasing trajectory of change in BW/BC when does are or not treated with melatonin. During natural anoestrus, 46 Blanca Andaluza does were assigned to two groups: 1) low BW/low BC group in which does were fed 1.9 times maintenance requirements for dietary energy for gaining BW/BC (LLg group; n = 23); or 2) a high BW/high BC group in which the does were fed 0.4 times maintenance requirements for dietary energy that resulted in a loss of BW/BC (HHl group; n = 23). There were similar numbers of does in each group that were treated or not treated with melatonin (MEL). Following 48 days of isolation from bucks, four sexually active individuals fitted with marking harnesses were transferred to the paddock containing the does of each group. Blood samples were collected by jugular venipuncture (before the distribution of concentrate) twice per week. The effect of the treatments (increasing or decreasing BW/BC and melatonin) on the different variables that were assessed were analysed using an ANOVA or the Fisher-Freeman-Halton exact probability test as necessary. During the 35 days after treatments were applied, the percentage of females expressing oestrous and having an ovulation were greater in the LLg + MEL than HHl-MEL subgroup (P < 0.05). The interaction of nutrition × melatonin treatment had a significant effect on reproduction of does (P < 0.05). This could be explained by the greater plasma glucose and IGF-1 and lesser plasma non-esterified fatty acid concentrations in does with increasing BW/BC (P < 0.01), and the greater IGF-1 concentrations of MEL-treated females (P < 0.01). The LH concentration and pulsatile release of this hormone from the pituitary were also modified by the presence of the males (P < 0.01). Furthermore, the LLg + MEL-treated does were responsive to the presence of bucks (P < 0.05). The present results indicate sexually active males cannot induce an adequate reproductive response in females with decreasing BW/BC even when does are being treated with melatonin. The presence of bucks enhanced the doe reproductive response when does were treated with melatonin and a pattern of increasing BW/BC.

Role of TrkA signalling and mast cells in the initiation of osteoarthritis pain in the monoiodoacetate model.

OBJECTIVE: Aiming to delineate novel neuro-immune mechanisms for NGF/TrkA signalling in osteoarthritis (OA) pain, we evaluated inflammatory changes in the knee joints following injection of monoiodoacetate (MIA) in mice carrying a TrkA receptor mutation (P782S; TrkA KI mice). METHOD: In behavioural studies we monitored mechanical hypersensitivity following intra-articular MIA and oral prostaglandin D2 (PGD2) synthase inhibitor treatments. In immunohistochemical studies we quantified joint mast cell numbers, calcitonin gene-related peptide expression in synovia and dorsal root ganglia, spinal cord neuron activation and microgliosis. We quantified joint leukocyte infiltration by flow cytometry analysis, and PGD2 generation and cyclooxygenase-2 (COX-2) expression in mast cell lines by ELISA and Western blot. RESULTS: In TrkA KI mice we observed rapid development of mechanical hypersensitivity and amplification of dorsal horn neurons and microglia activation 7 days after MIA. In TrkA KI knee joints we detected significant leukocyte infiltration and mast cells located in the vicinity of synovial nociceptive fibres. We demonstrated that mast cells exposure to NGF results in up-regulation of COX-2 and increase of PGD2 production. Finally, we observed that a PGD2 synthase inhibitor prevented MIA-mechanical hypersensitivity in TrkA KI, at doses which were ineffective in wild type (WT) mice. CONCLUSION: Using the TrkA KI mouse model, we delineated a novel neuro-immune pathway and suggest that NGF-induced production of PGD2 in joint mast cells is critical for referred mechanical hypersensitivity in OA, probably through the activation of PGD2 receptor 1 in nociceptors: TrkA blockade in mast cells constitutes a potential target for OA pain.

Long-term effects of hydrogen sulfide on the anabolic-catabolic balance of articular cartilage in vitro.

Healthy cartilage maintenance relies on an equilibrium among the anabolic and catabolic processes in chondrocytes. With the onset of osteoarthritis (OA), increased interleukin (IL)-1ß levels induce an inhibition of the synthesis of extracellular matrix (ECM) proteins, as well as an increase in proteases. This eventually leads to a predominance of the catabolic phenotype and the progressive loss of articular cartilage. Hydrogen sulfide (H2S) is a small gaseous molecule recognized as the third endogenous gasotransmitter. When administered exogenously, it has shown anti-inflammatory and anti-catabolic properties in several in vitro and in vivo models. Here, OA cartilage disks were co-cultured in vitro with IL-1ß (5 ng/ml) and NaSH or GYY4137 (200 or 1000 µM) for 21 days. The ability of these two H2S-producing compounds to avoid long term extracellular matrix (ECM) destruction was evaluated. We used a glycosaminoglycan (GAG) quantification kit histology and immunohistochemistry (IHC) to evaluate matrix proteins degradation and matrix metalloproteinases (MMP) abundance. Through the GAGs quantification assay, safranin O (S-O) and toluidine blue (TB) stains, and keratan/chondroitin sulfate (KS/ChS) IHCs it was shown that co-stimulation with H2S-forming reagents effectively avoided GAGs destruction. Both Masson's trichrome (MT) stain and collagen (col) type II IHC, as well as aggrecan (agg) IHC demonstrated that not only were these proteins protected but even promoted, their abundance being higher than in the basal condition. Further, stains also demonstrated that positivity in the inter-territorial and intra-cellular for the different matrix components were rescued, suggesting that NaSH and GYY4137 might also have pro-anabolic effects. In addition, a clear protective effect against the increased MMPs levels was seen, since increased MMP3 and 13 levels were subsequently reduced with the co-stimulation with sulfide compounds. In general, GYY4137 was more effective than NaSH, and increasing the dose improved the results. This study demonstrates that H2S anti-catabolic effects, which had been previously proven in short-term (24-48 h) in vitro cellular models, are maintained over time directly in OA cartilage tissue.

The isolation of females from males to promote a later male effect is unnecessary if the bucks used are sexually active.

It has been suggested that female goats in permanent contact with males become refractory to their presence, and need to be previously separated from them for 40-45 days if the presence of bucks is to induce reproductive activity, ovulation and oestrous during seasonal anoestrous. The present study examines the reproductive response (ovulation and oestrus) and reproductive performance of does isolated from bucks for different periods before their reintroduction to male company. A total of 103 Payoya and Blanca Andaluza does were distributed into six treatment groups that required their isolation from males for different periods: 0 days (N = 29), 5 days (N = 15), 10 days (N = 14), 20 days (N = 16), 30 days (N = 14) and 39 days (N = 15). After this period they were introduced to sexually active bucks (ensured to be in this condition by keeping them under long days light treatment for three months), and oestrous activity was recorded daily by direct visual observation of the marks left by the marking harnesses worn by these males. Ovulation was confirmed via the plasma progesterone concentration (measured in blood samples taken twice per week). The ovulation rate was assessed by transrectal ultrasonography. Fecundity, fertility, prolificacy and productivity were also determined. The sexual behaviour of the males towards the females was also monitored on Days 0, 1, 2, 3, 4, 8 and 9 after their meeting with the latter. The length of the female isolation period had no effect on the percentage of does that responded to contact with the males, nor did it affect the oestrous response, fecundity, fertility or productivity. The males, however, undertook more ano-genital sniffing and nudging with the 5 day group females compared to those of the other groups (P < 0.05). However, the sexual behaviour of the males changed as the days passed, with ano-genital sniffing becoming less common, and nudging, licking, sneezing and mounts with intromission more frequent on Days 8 and 9 than on Day 0, 1 and 2 after the sexes were reunited (P < 0.05). These results show that the isolation of females is not necessary for an efficient male effect if the bucks used are sexually active. In addition, the sexual behaviour of the bucks changes as the time in contact with the does increases, but in general is not affected by the duration of female isolation.