Caesalpinia palmeri: First Report on the Phenolic Compounds Profile, Antioxidant and Cytotoxicity Effect.

This study aimed to determine the phenolic compounds profile, antioxidant potential and cytotoxicity of extracts and fractions of Caesalpinia palmeri. Methanolic extracts were generated from C. palmeri berries, stems and flowers. The latter was subjected to liquid-liquid partition, obtaining hexane, ethyl acetate and residues fractions. Results showed that the flower extract and ethyl acetate fraction had a larger concentration of phenolic compounds (148.9 and 307.9 mg GAE/g, respectively), being ellagic acid (6233.57 and 19550.08 µg/g, respectively), quercetin-3-ß-glycoside (3023.85 and 8952.55 µg/g, respectively) and gallic acid (2212.98 and 8422.34 µg/g, respectively) the most abundant compounds. Flower extract and ethyl acetate fraction also presented the highest antioxidant capacity on all tested methods (DPPH, ABTS, ORAC and FRAP) and low cytotoxicity against ARPE-19 cells (IC50 >170 µg/mL). C. palmeri possessed high antioxidant potential, associated with the presence of phenolic compounds and low cytotoxicity, suggesting that they could represent an option to counter oxidative stress.

Evaluation and comparison of selected methodologies to investigate occupational accidents.

BACKGROUND: The choice of the "right" methodology to carry out the occupational accident investigation process is not an easy task. Each methodology has different conceptual and practical characteristics. The choice will depend to a large extent on the conceptual accident model being used. OBJECTIVE: The aim of this study has been to evaluate and compare a set of nine methodologies recognised as the most widely used in the field of occupational accident investigation. METHODS: For this purpose, six evaluation criteria are used which have already been applied and validated by the scientific community with a clear emphasis on the holistic nature of these methodologies. RESULTS: The results show a detailed analysis of the conceptual, methodological and practical characteristics of the nine selected occupational accident investigation methodologies. The conceptual framework of each of the methodologies, their holistic characteristics in terms of whether they cover the complete information cycle and its possible interrelation, reliability and validity of the methodologies, the experience required for their application, the flexibility in terms of being able to be used in different work environments and finally the ability of the methodology to motivate organisational improvement are presented. CONCLUSIONS: From this study, it is clear that the analysis of occupational accident investigation it is becoming increasingly necessary to employ scientific methodologies with a clear proactive approach in order to meet the challenges of changing socio-technical systems.

Effects of sub-optimal illumination in plants. Comprehensive chlorophyll fluorescence analysis.

The fluorescence signals emitted by chlorophyll molecules of plants is a promising non-destructive indicator of plant physiology due to its close link to photosynthesis. In this work, a deep photophysical study of chlorophyll fluorescence was provided, to assess the sub-optimal illumination effects on three plant species: L. sativa, A. hybridus and S. dendroideum. In all the cases, low light (LL) treatment induced an increase in pigment content. Fluorescence ratios - corrected by light reabsorption processes - remained constant, which suggested that photosystems stoichiometry was conserved. For all species and treatments, quantum yields of photophysical decay remained around 0.2, which meant that the maximum possible photosynthesis efficiency was about 0.8. L. sativa (C3) acclimated to low light illumination, displayed a strong increase in the LHC size and a net decrease in the photosynthetic efficiency. A. hybridus (C4) was not appreciably stressed by the low light availability whereas S. dendroideum (CAM), decreased its antenna and augmented the quantum yield of primary photochemistry. A novel approach to describe NPQ relaxation kinetics was also presented here and used to calculate typical deactivation times and amplitudes for NPQ components. LL acclimated L. sativa presented a much larger deactivation time for its state-transition-related quenching than the other species. Comprehensive fluorescence analysis allowed a deep study of the changes in the light-dependent reactions of photosynthesis upon low light illumination treatment.

Identifying the incidence of respiratory complications following diaphragmatic cytoreduction and hyperthermic intraoperative intraperitoneal chemotherapy.

INTRODUCTION: In patients with peritoneal carcinomatosis (PC), the incidence of respiratory complications following cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) is not well established. We aimed to describe the center-specific incidence and patient characteristics associated with respiratory complications following CRS and HIPEC in patients receiving treatment for PC. MATERIALS AND METHODS: We used the University Hospital of Arrixaca study database to identify patients who underwent CRS and HIPEC for PC. Patients who experienced a post-operative respiratory complication were categorized according to the National Cancer Institute-Common Terminology Criteria for Adverse Events. Multivariable regression methods were used to identify independent risk factors for developing a respiratory complication following CRS and HIPEC. RESULTS: Between 2008 and 2017, we identified 247 patients who underwent CRS and HIPEC for PC. A total of eight patients (3.2%) were categorized as having a post-operative respiratory complication. A diaphragmatic peritonectomy and a PC index of > 14 were identified as independent risk factors for developing a respiratory complication. Radiographic evidence of a pleural effusion was identified in 72 patients who had CRS of the diaphragmatic peritoneum; however, only 6 (8.3%) of these patients required pleural drainage. CONCLUSIONS: Only 3.2% of patients developed a symptomatic respiratory complication following CRS and HIPEC. A pleural effusion was identified in almost all patients requiring a diaphragmatic peritonectomy as part of their CRS; however, less than one in ten of these patients required pleural drainage. Prophylactic insertion of a pleural drainage tube is, therefore, not indicated following CRS and HIPEC.

Chemical composition of industrially and laboratory processed Cyperus esculentus rhizomes.

We report here the results of the study of the chemical composition of Cyperus esculentus rhizomes. Ethanolic extracts have been separated by column chromatography and analyzed by magnetic resonance spectroscopy and mass spectrometry. Quercetin, stigmasterol, and linoleic and oleic acid glycerol esters, together with 4-chlorobutyl oleate, oleamide, myricetin, tyramine and N-feruloyltyramine, found for the first time in Cyperus esculentus rhizomes, have been isolated and quantified in the extracts. Alkaloids have not been detected, and the presence of flavonoids and sterols is moderate.

Prisoner rehabilitation through animal-assisted activities in Argentina: the Huellas de Esperanza Prison Dog Programme. / Resocialización de personas privadas de libertad a través de actividades con animales en Argentina: el programa «Huellas de Esperanza¼.

One of the most traumatic events that a person can ever experience in their lifetime is being detained. The prison dog programmes are the most successful prison rehabilitation programmes in the United States of America and consist of detainees training service dogs for other people to assist with different types of disability. It is a typical win-win system: inmates win as they connect with their deepest feelings of affection and solidarity while acquiring a concrete skill for their future release; people with disabilities win because they receive a dog to support their day-to-day activities; the dogs win (especially adopted street dogs) because their lot in life is improved; institutions win because they are enabled to meet their objective more fully; and society wins because they gain a concrete tool to aid the social rehabilitation of detainees. This article describes the operation and achievements of Huellas de Esperanza (Traces of Hope), the Argentine prison dog programme, which has been implemented by the Federal Penitentiary Service since 2010 using State resources. A total of 85 inmates have participated in the programme, which has delivered more than a dozen assistance dogs to people with disabilities and many more dogs to elderly people to provide them with affection and companionship. The article also describes the programme methodology and the way it has decreased prison unrest and led to a low reoffending rate among programme participants.

La privation de liberté est l'une des situations les plus traumatisantes qu'un être humain puisse connaître au cours de son existence. De tous les programmes de réhabilitation appliqués dans les prisons aux États-Unis d'Amérique, Prison Dog Programme est celui qui donne les meilleurs résultats ; il consiste à confier aux détenus la tâche de dresser des chiens en vue d'en faire des chiens aidants qui rendront des services à d'autres personnes atteintes de divers types de handicap. Il s'agit d'un système typiquement « gagnant-gagnant ¼ : le détenu est gagnant, car il renoue avec des sentiments positifs d'affection et de solidarité, tout en recevant un outil concret pour sa vie future une fois libéré ; la personne handicapée est gagnante, car elle reçoit un chien qui l'aidera à vivre au quotidien ; le chien est gagnant, surtout s'il était un chien errant avant d'être adopté, car il accède à une vie meilleure ; enfin, les institutions sont gagnantes car leurs objectifs sont mieux remplis, de même que la société, qui grâce à cet outil concret facilite, la réinsertion sociale des personnes privées de liberté. Les auteurs décrivent le fonctionnement et les réussites de la version argentine de ce programme, intitulée « Traces d'espérance ¼ (Huellas de Esperanza). Le programme a été mis en place par le système pénitentiaire fédéral en 2010 et doté de véritables ressources étatiques; au total, 86 détenus en ont bénéficié et plus d'une douzaine de chiens aidants ont été remis à des personnes handicapées, en plus du grand nombre de chiens dressés et remis à des centres de gériatrie pour apporter un réconfort aux pensionnaires. Les auteurs expliquent la méthodologie suivie, ainsi que la réduction des conflits en milieu carcéral et le faible taux de récidives parmi les ex-détenus ayant participé au programme.

Una de las situaciones más traumáticas que puede atravesar un ser humano a lo largo de su existencia es la privación de su libertad. Los Prison Dog Programmes son los sistemas de rehabilitación carcelaria más exitosos de EE.UU., y consisten en el adiestramiento de perros por parte de personas privadas de la libertad, que los preparan como perros de servicio para apoyo de distintos tipos de discapacidad en otras personas. El sistema es un típico win-win-win: gana el interno conectándose con sus mejores sentimientos de afecto y solidaridad al tiempo que recibe una herramienta concreta para su futuro en libertad; gana la persona con discapacidad que recibe un perro como apoyo para su desempeño cotidiano; gana el perro, sobre todo si se trata de perros vagabundos adoptados que obtienen un destino mejor; ganan las instituciones, que cumplen mejor su objetivo, y gana la sociedad, que facilita, con una herramienta concreta, la reinserción social de las personas privadas de libertad. El presente trabajo describe la operatoria y logros de Huellas de Esperanza, la versión argentina de este Programa, que se desarrolla en el Servicio Penitenciario Federal desde 2010 con recursos estatales genuinos; por este programa han pasado más de 85 internos y ha permitido entregar más de una docena de perros de asistencia a personas con discapacidad y muchos perros a geriátricos para compañía afectiva. Se describe también la metodología y la disminución de la conflictividad intracarcelaria, así como la escasa reiteración de sus participantes.

Fatty acid profile of goat milk in diets supplemented with chia seed (Salvia hispanica L.).

Chia seed (Salvia hispanica L.) is the greatest known plant source of n-3 α-linolenic acid. The present study evaluated the effects of 3 inclusion levels of chia seed [zero (control); low, 2.7% (CLow); and high, 5.5% (CHigh)] in diets of dairy goats on milk yield and fatty acid profile. Nine Saanen dairy goats in the last third of lactation period, live weight 38 ± 8.7 kg, housed in metabolic cages, were fed iso-proteic and iso-energetic (160 g of crude protein/d and 11 MJ of metabolizable energy/d) diets. Gas chromatography was used to analyze fatty acid profile and total conjugated linoleic acid (CLA). Silver ion HPLC was used to analyze the isomeric profile of CLA. The results were subjected to variance analysis using a Latin square design repeated 3 × 3. The CHigh treatment was higher for dry matter, neutral detergent fiber, and acid detergent fiber intake compared with CLow and control diets. Digestibility was not affected by the inclusion of chia seeds. The CHigh diet improved N intake with respect to the control and CLow diet. Milk yield and chemical composition were not affected by the treatment. The milk fatty acid profile of C18:0, C18:1, C18:2, and C:20 was higher for CHigh than the other treatments. The in vitro gas production (mL of gas/g of dry matter) was lower in CHigh than the control diet. In conclusion, the addition of chia seeds at the CHigh level in dairy goat diets negatively affected in vitro rumen fermentation, but increased the milk fatty acid profile of C18:0, C18:1n-9 cis, and C:20, monounsaturated fatty acids, and polyunsaturated fatty acids. The total CLA content increased from 0.33 to 0.73% with the supplementation of chia to the diet, as well as the isomers cis-9,trans-11, trans-7,cis-9, trans-11,cis-13, and trans-12,trans-14.

Prevalence of oral allergy syndrome in children with allergic diseases.

INTRODUCTION: The oral allergy syndrome (OAS) is a particular type of food allergy rarely explored in the paediatric population that is already considered an adult problem. OBJECTIVE: Identify the prevalence of OAS, symptoms and pollen species associated with its presence in children affected by allergic diseases. METHODS: A cross-sectional study was conducted. Consecutive sampling included children from 6 to 14 years who needed allergy treatment for the first time. A structured questionnaire was carried out to collect demographic and clinical data and history of OAS. Besides sensitisation to various allergens, the skin prick-by-prick test was performed to corroborate sensitisation to food related to OAS. Prevalence of OAS and its association with pollens was established following the covariate adjusted logistic regression. RESULTS: 267 subjects were included. Overall prevalence of OAS was 8.9% (95%CI 6.1-13.1%). Prevalence of OAS for allergic rhinitis and asthma were 8.8% and 9.1%, respectively. In patients sensitised to pollen, the prevalence ranged from 9.6% to 12.2% depending on the type of pollen. 62.5% of children with OAS were sensitive to pineapple. After adjusting for gender and family history of atopic disease, trees from the Quercus species showed an association with OAS (OR=2.7, 95%CI 1.2-6.2). CONCLUSIONS: OAS is not uncommon in our environment. Pineapple, a typical fruit from the region, was the main food related. Quercus sp., but not birch nor olive, was the pollen associated with this syndrome.

The effects of prebiotics on microbial dysbiosis, butyrate production and immunity in HIV-infected subjects.

Altered interactions between the gut mucosa and bacteria during HIV infection seem to contribute to chronic immune dysfunction. A deeper understanding of how nutritional interventions could ameliorate gut dysbiosis is needed. Forty-four subjects, including 12 HIV+ viremic untreated (VU) patients, 23 antiretroviral therapy-treated (ART+) virally suppressed patients (15 immunological responders and 8 non-responders) and 9 HIV- controls (HIV-), were blindly randomized to receive either prebiotics (scGOS/lcFOS/glutamine) or placebo (34/10) over 6 weeks in this pilot study. We assessed fecal microbiota composition using deep 16S rRNA gene sequencing and several immunological and genetic markers involved in HIV immunopathogenesis. The short dietary supplementation attenuated HIV-associated dysbiosis, which was most apparent in VU individuals but less so in ART+ subjects, whose gut microbiota was found more resilient. This compositional shift was not observed in the placebo arm. Significantly, declines in indirect markers of bacterial translocation and T-cell activation, improvement of thymic output, and changes in butyrate production were observed. Increases in the abundance of Faecalibacterium and Lachnospira strongly correlated with moderate but significant increases of butyrate production and amelioration of the inflammatory biomarkers soluble CD14 and high-sensitivity C-reactive protein, especially among VU. Hence, the bacterial butyrate synthesis pathway holds promise as a viable target for interventions.

Effects of maturity at ensiling of bermudagrass and fibrolytic enzyme application on the performance of early-lactation dairy cows.

The objective of this study was to examine effects of adding fibrolytic enzymes to diets containing bermudagrass ensiled after 4 or 7wk of regrowth on the diet digestibility, ruminal fermentation and performance of lactating cows, and the interaction of the treatments. In experiment 1, 64 Holstein cows (22±4d in milk) were assigned to an experiment with a 2×2 factorial treatment arrangement and a 56-d duration. Treatments were diets containing 4 or 7wk regrowth bermudagrass silage without or with an exogenous fibrolytic enzyme cocktail. The cellulase-xylanase enzyme was applied at 2.33g/kg of total mixed ration dry matter (DM) during mixing immediately before feeding. Experiment 2 was aimed at examining treatment effects on the ruminal fermentation profile. Four ruminally cannulated cows were assigned to the 4 treatments using a 4×4 Latin square design with 14-d periods. No enzyme by maturity interaction was detected for any measurement. Regardless of forage maturity, applying the fibrolytic enzyme did not affect DM intake, milk yield, apparent digestibility, feed efficiency, energy balance, and ruminal fermentation though it tended to increase milk lactose concentration (4.88 vs. 4.81%). Feeding the 4-wk diet instead of the 7-wk diet increased DM intake (22.4 vs. 21.3kg/d), digestibility of DM, neutral detergent fiber, and acid detergent fiber, and tended to increase 3.5%-fat corrected milk yield (47.2 vs. 44.3kg/d) and milk fat yield (1.88 vs. 1.73kg/d). Therefore, daily intake of net energy and secretion of milk energy were greater for the 4-wk diet. In addition, the 4-wk diet increased the ruminal concentrations of acetate, propionate, valerate, lactate, and total volatile fatty acids, and decreased ruminal pH, without affecting the acetate:propionate ratio. Feeding fibrolytic enzymes did not improve the performance of early-lactation dairy cows, but harvesting the forage earlier tended to improve milk production.

Quantitative analysis of ruminal bacterial populations involved in lipid metabolism in dairy cows fed different vegetable oils.

Vegetable oils are used to increase energy density of dairy cow diets, although they can provoke changes in rumen bacteria populations and have repercussions on the biohydrogenation process. The aim of this study was to evaluate the effect of two sources of dietary lipids: soybean oil (SO, an unsaturated source) and hydrogenated palm oil (HPO, a saturated source) on bacterial populations and the fatty acid profile of ruminal digesta. Three non-lactating Holstein cows fitted with ruminal cannulae were used in a 3×3 Latin square design with three periods consisting of 21 days. Dietary treatments consisted of a basal diet (Control, no fat supplement) and the basal diet supplemented with SO (2.7% of dry matter (DM)) or HPO (2.7% of DM). Ruminal digesta pH, NH3-N and volatile fatty acids were not affected by dietary treatments. Compared with control and HPO, total bacteria measured as copies of 16S ribosomal DNA/ml by quantitative PCR was decreased (P<0.05) by SO. Fibrobacter succinogenes, Butyrivibrio proteoclasticus and Anaerovibrio lipolytica loads were not affected by dietary treatments. In contrast, compared with control, load of Prevotella bryantii was increased (P<0.05) with HPO diet. Compared with control and SO, HPO decreased (P<0.05) C18:2 cis n-6 in ruminal digesta. Contents of C15:0 iso, C18:11 trans-11 and C18:2 cis-9, trans-11 were increased (P<0.05) in ruminal digesta by SO compared with control and HPO. In conclusion, supplementation of SO or HPO do not affect ruminal fermentation parameters, whereas HPO can increase load of ruminal P. bryantii. Also, results observed in our targeted bacteria may have depended on the saturation degree of dietary oils.

Improving the performance of dairy cattle with a xylanase-rich exogenous enzyme preparation.

The objective of this experiment was to examine effects of adding 2 exogenous fibrolytic enzymes (EFE) to the total mixed ration (TMR) on the performance of lactating dairy cows (experiment 1) and the kinetics of ruminal degradation of the diet (experiment 2). Twelve EFE had been screened in a series of in vitro assays that identified the most potent EFE and their optimal doses for increasing the digestibility of bermudagrass. In experiment 1, 66 Holstein cows (21±5 d in milk) were grouped by previous milk production and parity (45 multiparous and 21 primiparous) and assigned randomly to 1 of the following 3 treatments: (1) control (CON, untreated), (2) Xylanase Plus [2A, 1mL/kg of TMR dry matter (DM); Dyadic International, Jupiter, FL], and (3) a 75:25 (vol/vol) mixture of Cellulase Plus and Xylanase Plus EFE (3A, 3.4mL/kg of TMR DM; Dyadic International). The EFE were sprayed twice daily onto a TMR (10% bermudagrass silage, 35% corn silage, 5% alfalfa-orchardgrass hay mixture, and 50% concentrates; DM basis) and fed for a 14-d training and covariate period and a 70-d measurement period. Experiment 2 aimed to examine the in situ DM ruminal degradability and ruminal fermentation measurements of the diets fed in experiment 1. Three ruminally fistulated lactating Holstein cows were assigned to the diets. The experiment had a 3×3 Latin square design with 23-d periods. In experiment 1, application of 2A increased intakes (kg/d) of DM (23.5 vs. 22.6), organic matter (21.9 vs. 20.9), and crude protein (3.9 vs. 3.7) and tended to increase yields (kg/d) of fat-corrected milk (41.8 vs. 40.7) and milk fat (1.48 vs. 1.44). In particular, 2A increased milk yield (kg/d) during wk 3 (41.2 vs. 39.8, tendency), 6 (41.9 vs. 40.1), and 7 (42.1 vs. 40.4), whereas 3A increased milk yield (kg/d) during wk 6 (41.5 vs. 40.1, tendency), 8 (41.8 vs. 40.0), and 9 (40.9 vs. 39.5, tendency). In experiment 2, EFE treatment did not affect ruminal DM degradation kinetics or ruminal pH, ammonia-N, and volatile fatty acid concentration. Application of 2A to the bermudagrass-based TMR increased DM intake and milk production, implying that this EFE could be used to increase the performance of lactating dairy cows fed diets containing up to 10% bermudagrass.

Effect of adding cofactors to exogenous fibrolytic enzymes on preingestive hydrolysis, in vitro digestibility, and fermentation of bermudagrass haylage.

Our objectives were to examine if adding metal ion cofactors (COF) to exogenous fibrolytic enzymes (EFE) would increase the beneficial effects of the EFE on the preingestive hydrolysis and in vitro digestibility and fermentation of bermudagrass haylage. In experiment 1, 5 COF (Mn(2+), Co(2+), Fe(2+), Ca(2+), and Mg(2+)) were screened to select the best candidates for synergistically enhancing release of water-soluble carbohydrates (WSC) from bermudagrass haylage by 5 EFE. The 5 EFE (1A, 2A, 11C, 13D, and 15D) were sourced from Trichoderma reesei and Aspergillus oryzae and they were the most effective of 12 EFE at increasing the neutral detergent fiber digestibility of bermudagrass haylage in a previous trial. Adding 1mM of each of the COF to EFE 2A or 11C synergistically increased release of WSC from bermudagrass haylage, as did adding (1mM) Fe(2+) to 1A, Mn(2+), Co(2+), or Fe(2+) to 13D, or Co(2+)or Fe(2+) to 15D. The greatest release of WSC responses were obtained by adding Mn(2+) to 11C (38%) or by adding Fe(2+) to 2A or 13D (10 and 21.9%, respectively). In experiment 2, the effect of increasing the COF dose on in vitro digestibility and fermentation of bermudagrass haylage was examined using the best EFE-COF combinations from experiment 1. Effects of adding increasing doses of these COF on EFE-mediated changes in vitro digestibility depended on the COF-EFE combination. Adding 10mM Mn(2+) alone to bermudagrass haylage increased DMD and NDFD by 2.7 and 6.3% and adding 11C alone increased these measures by 6.6 and 15.5%, respectively. However, adding 10mM Mn(2+) with 11C resulted in 3.5 and 8.1% increases in DMD and NDFD, respectively, beyond the increases caused by adding 11C alone. Adding Fe(2+) to 2A had no effects on EFE-mediated digestibility responses, but 2A prevented adverse effects of adding Fe(2+) alone on DMD and NDFD. In contrast, adding Fe(2+) to 13D reduced the increases in DMD and NDFD caused by adding the EFE alone. This study shows that adding COF to EFE can synergistically increase, decrease, or not affect the hydrolytic effects of EFE on bermudagrass haylage cell walls. The outcome depends on the specific EFE-COF combination and the COF dose. More research is required to understand the mechanisms resulting in these outcomes to exploit beneficial effects of COF on EFE.

Screening exogenous fibrolytic enzyme preparations for improved in vitro digestibility of bermudagrass haylage.

Our objectives were to evaluate the effects of 12 exogenous fibrolytic enzyme products (EFE) on ruminal in vitro neutral detergent fiber digestibility (NDFD) and preingestive hydrolysis of a 4-wk regrowth of bermudagrass haylage (BH), to examine the accuracy of predicting NDFD with EFE activity measures, and to examine the protein composition of the most and least effective EFE at increasing NDFD. In experiment 1, effects of 12 EFE on NDFD of BH were tested. Enzymes were applied in quadruplicate to culture tubes containing ground BH. The suspension was incubated for 24 h at 25 °C before addition of rumen fluid media and further incubation for 24 h at 39 °C. The experiment was repeated twice. In addition, regression relationships between EFE activity measures and NDFD were examined. Compared with the values for the control, 9 EFE-treated substrates had greater NDFD (37.8 to 40.4 vs. 35.6%), 6 had greater total VFA concentration (59.1 to 61.2 vs. 55.4 mM), and 4 had lower acetate-to-propionate ratios (3.03 to 3.16 vs. 3.24). In experiment 2, EFE effects on preingestive fiber hydrolysis were evaluated by incubating enzyme-treated and untreated bermudagrass suspensions in quadruplicate for 24 h at 25 °C and examining fiber hydrolysis measures. Compared with values for the control, 3 EFE reduced neutral detergent fiber concentration (62.8 to 63.7 vs. 67.3%), 10 increased release of water-soluble carbohydrates (26.8 to 58.5 vs. 22.8 mg/g), and 8 increased release of ferulic acid (210 to 391 vs. 198 µg/g). Regression analyses revealed that enzyme activities accurately [coefficient of determination (R(2)) = 0.98] predicted preingestive hydrolysis measures (water-soluble carbohydrates, ferulic acid), moderately (R(2) = 0.47) predicted neutral detergent fiber hydrolysis, but poorly (R(2) ≤ 0.1) predicted dry matter and NDFD. In experiment 3, proteomic tools were used to examine the protein composition of the most and least effective EFE at improving NDFD. Relative to the least effective, the most effective EFE at increasing NDFD contained 10 times more endoglucanase III, 17 times more acetylxylan esterase with a cellulose-binding domain 1, 33 times more xylanase III, 25 times more ß-xylosidase, and 7.7 times more polysaccharide monooxygenase with cellulose-binding domain 1 and 3 times more swollenin. The most effective EFE had a much greater quantity of fibrolytic enzymes and key proteins necessary for hemicellulose and lignocellulase deconstruction. This study identified several EFE that increased the NDFD and in vitro fermentation of 4-wk BH and revealed why some EFE are more effective than others.

Effect of the dose of exogenous fibrolytic enzyme preparations on preingestive fiber hydrolysis, ruminal fermentation, and in vitro digestibility of bermudagrass haylage.

Our objectives were to evaluate the effects of the dose rates of 5 Trichoderma reesei and Aspergillus oryzae exogenous fibrolytic enzymes (EFE; 1A, 2A, 11C, 13D, and 15D) on in vitro digestibility, fermentation characteristics, and preingestive hydrolysis of bermudagrass haylage and to identify the optimal dose of each EFE for subsequent in vitro and in vivo studies. In experiment 1, EFE were diluted in citrate-phosphate buffer (pH 6) and applied in quadruplicate in each of 2 runs at 0× (control), 0.5×, 1×, 2×, and 3×; where 1× was the respective manufacturer-recommended dose (2.25, 2.25, 10, 15, and 15g of EFE/kg of dry matter). The suspension was incubated for 24h at 25°C and for a further 24h at 39°C after the addition of ruminal fluid. In experiment 2, a similar approach to that in experiment 1 was used to evaluate simulated preingestive effects, except that sodium azide (0.02% wt/vol) was added to the EFE solution. The suspension was incubated for 24h at 25°C and then 15mL of water was added before filtration to extract water-soluble compounds. For both experiments, data for each enzyme were analyzed separately as a completely randomized block design with a model that included effects of EFE dose, run, and their interaction. In experiment 1, increasing the EFE dose rate nonlinearly increased the DM digestibility of 1A, 2A, 11C, and 13D and the neutral detergent fiber digestibility (NDFD) of 1A, 2A, 11C, and 13D. Optimal doses of 1A, 2A, 11C, 13D, and 15D, as indicated by the greatest increases in NDFD at the lowest dose tested, were 2×, 2×, 1×, 0.5×, and 0.5×, respectively. Increasing the dose rate of 2A, 11C, and 13D nonlinearly increased concentrations of total volatile fatty acids and propionate (mM), decreased their acetate-to-propionate ratios and linearly decreased those of samples treated with 1A and 15D. In experiment 2, increasing the dose rate of each EFE nonlinearly decreased concentrations of netural detergent fiber; also, increasing the dose rate of 1A, 2A, 11C, and 1D nonlinearly increased concentrations of water-soluble carbohydrates and free ferulic acid (µg/g). Application of increasing doses of the EFE increased NDF hydrolysis, NDFD, and ruminal fluid fermentation of bermudagrass haylage, but the optimal dose varied with the EFE.

Effect of microbial inoculants on the quality and aerobic stability of bermudagrass round-bale haylage.

The objective of this study was to compare the efficacy of using 4 commercially available microbial inoculants to improve the fermentation and aerobic stability of bermudagrass haylage. We hypothesized that the microbial inoculants would increase the fermentation and aerobic stability of the haylages. Bermudagrass (4-wk regrowth) was harvested and treated with (1) deionized water (control); (2) Buchneri 500 (B500; Lallemand Animal Nutrition, Milwaukee, WI) containing 1×10(5) of Pediococcus pentosaceus and 4×10(5) of Lactobacillus buchneri 40788; (3) Biotal Plus II (BPII; Lallemand Animal Nutrition) containing 1.2×10(5) of P. pentosaceus and Propionibacteria freudenreichii; (4) Silage Inoculant II (SI; AgriKing Inc., Fulton, IL) containing 1×10(5) of Lactobacillus plantarum and P. pentosaceus; and (5) Silo King (SK; AgriKing Inc.), containing 1×10(5) of L. plantarum, Enterococcus faecium, and P. pentosaceus, respectively. Forty round bales (8 per treatment; 441±26kg; 1.2×1.2 m diameter) were made and each was wrapped with 7 layers of plastic. Twenty bales were stored for 112 d and the remaining 20 were stored for 30 d and sampled by coring after intermediary storage periods of 0, 3, 7, and 30 d. The pH of control and inoculated haylages sampled on d 3 did not differ. However, B500 and BPII had lower pH (5.77±0.04 vs. 6.16±0.04; 5.06±0.13 vs. 5.52±0.13) than other treatments by d 7 and 30, respectively. At final bale opening on d 112, all treatments had lower pH than the control haylage (4.77±0.07 vs. 5.37±0.07). The B500, BPII, and SI haylages had greater lactic acid and lactic-to-acetic acid ratios than SK and control haylages. No differences were detected in neutral detergent fiber digestibility, dry matter losses, dry matter, lactic and acetic acid concentrations, and yeast and coliform counts. The SK haylage had lower clostridia counts compared with the control (1.19±0.23 vs. 1.99±0.23 cfu/g). Treatments B500, BPII, SI, and SK tended to reduce mold counts and they improved aerobic stability by 236, 197, 188, and 95%, respectively, compared with the control (276±22 vs. 99±22h).

Cholesterol modulates LRP5 expression in the vessel wall.

OBJECTIVE: Macrophages are key players in atherosclerotic lesion formation and progression. We have recently demonstrated that lipid-loaded macrophages show activation of the canonical Wnt signaling pathway. METHODS: To test the in vivo role of the canonical Wnt pathway in atherosclerosis we used mice deficient in the Wnt signaling receptor LRP5 (LRP5(-/-)) fed a hypercholesterolemic diet (HC) to induce atherosclerosis. These dietary groups were further subdivided into two subgroups receiving their respective diets supplemented with 2% plant sterol esters (PSE). All mice remained on their assigned diets until age 18 weeks. RESULTS: HC WT mice had mildly increased non-HDL cholesterol levels, developed aortic atherosclerotic lesions and showed upregulated expression levels of aortic Lrp5. HC LRP5(-/-) mice develop larger aortic atherosclerotic lesions than WT mice indicating that LRP5 has a protective function in atherosclerosis progression. The oral administration of PSE, a dietary cholesterol-lowering agent, had an effect in the expression levels of the Wnt signaling receptor and in atherosclerosis progression. We found that PSE reduced serum total cholesterol levels, abolished HC-induced LRP5 overexpression and reduced aortic atherosclerotic plaques. CONCLUSION: The proatherogenic effects of the excess of plasma lipids are in part mediated by modulation of LRP5 in the aorta. LRP5 and canonical Wnt signaling exert a protective defense mechanism against hyperlipidemia and atherosclerosis lesion progression.