Oxidation of model lipids representative for main paper pulp lipophilic extractives by the laccase-mediator system.

Several model lipids representative for main paper pulp lipophilic extractives - including alkanes, fatty alcohols, fatty acids, resin acids, free and esterified sterols, and triglycerides – were treated with Pycnoporus cinnabarinus laccase in the presence of 1-hydroxybenzotriazole as mediator. The reaction products were analyzed by GC and GC-MS. The laccase alone decreased the amount of some unsaturated lipids, however, the most rapid and extensive lipid modification was obtained with the laccase-mediator system. Most unsaturated lipids were largely oxidized and the dominant oxidation products detected were epoxy and hydroxy-fatty acids from fatty acids, and free and esterified 7-ketosterols and steroid ketones from sterols and sterol esters. In contrast, saturated lipids were not modified, although some of them were oxidized when the enzymatic reactions were carried out in the presence of unsaturated lipids. The results obtained are discussed in the context of enzymatic control of pitch deposits, to explain the removal of lipid mixtures during laccase-mediator treatment of different pulp types.This study was funded by the BIORENEW EU-project (NMP2-CT-2006-026456) and the Spanish MEC (BIO2007-28719-E). Beldem (Andenne, Belgium) is acknowledged for laccase supply. S.M. thanks the Spanish CSIC and CELESA for an I3P contract and J.R thanks the CSIC for an I3P fellowship.Peer reviewe

A Wearable System for Multisensory Stimulation Therapy for Children

Multisensory stimulation therapy involves the simultaneous stimulation of several senses in a relaxing environment to achieve a variety of therapeutic outcomes for client with conditions affecting sensory and cognitive processes. We present, StimuHat, a wearable system for therapists to visually stimulate patients. We conducted a pilot study in which a therapist used StimuHat in sessions with three children with profound brain damage. The results showed that StimuHat appears to have stimulated the children and created positive relaxation and engagement in them

Influence of operation conditions on laccase-mediator removal of sterols from eucalypt pulp.

*Textile and Paper Engineering Department, ETSEIAT, Universitat Politècnica de Catalunya, Colom 11, E-08222 Terrassa, Spain. **Instituto de Recursos Naturales y Agrobiología, CSIC, PO Box 1052, E-41080 Seville, Spain ***Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, E-28040 Madrid, Spain; E-mail address: [email protected] (M.B. Roncero)The way how sterols, the main lipophilic compounds present in eucalypt kraft pulp, are eliminated by an enzymatic stage using the laccase-mediator system was evaluated. With this purpose laccase-mediator stage (L) was applied on an Eucalyptus globulus pulp under different operation conditions following a three-variable (laccase dose, mediator dose and reaction time) sequential statistical plan, to optimise the removal of sterols. The decrease in pulp sterol content during the enzymatic treatment was related to the decrease in kappa number and to brightness increase, as well as with the increase in some oxidation products of sitosterol (namely 7-oxositosterol and stigmasta-3,5-dien-7-one). The increase in reaction time from 1 to 5 h strongly reduced the sterol content, while no more sterols were eliminated during the 5-7 h period. Increasing the laccase dose from 1 to 20 U g-1 of pulp produced a high reduction in pulp sterols, whereas the increase in mediator (1-hydroxybenzotriazole) dose (from 0.5 to 2.5 % of pulp weight) had only a slight influence in removing sterols. Therefore, at 16 U g-1 laccase dose, 0.5% mediator dose, 4 h of reaction, practically all the sterols were removed. Finally, it was demonstrated that sterols were more sensitive to a L stage (practically 100% of sterols were eliminated) than to a chlorine dioxide stage (54% of sterols eliminated).This work was funded by the Spanish MEC (BIOFIBER project, CTQ2007-68003-C02-01; and project BIO2007-28719-E), MICINN (ELLE project, AGL2008-00709) and the European Commission (project BIORENEW, NMP2-CT-2006-026456). Torraspapel S.A. (Zaragoza, Spain) and Novozymes (Bagsvaerd, Denmark) are gratefully acknowledged for supplying the pulp and enzymes used, respectively.Peer reviewe

Fasciola hepatica induces eosinophil apoptosis in the migratory and biliary stages of infection in sheep

The aim of the present work was to evaluate the number of apoptotic eosinophils in the livers of sheep experimentally infected with Fasciola hepatica during the migratory and biliary stages of infection. Four groups (n = 5) of sheep were used; groups 1–3 were orally infected with 200 metacercariae (mc) and sacrificed at 8 and 28 days post-infection (dpi), and 17 weeks post-infection (wpi), respectively. Group 4 was used as an uninfected control. Apoptosis was detected using immunohistochemistry with a polyclonal antibody against anti-active caspase-3, and transmission electron microscopy (TEM). Eosinophils were identified using the Hansel stain in serial sections for caspase-3, and by ultrastructural features using TEM. At 8 and 28 dpi, numerous caspase-3+ eosinophils were mainly found at the periphery of acute hepatic necrotic foci. The percentage of caspase -3+ apoptotic eosinophils in the periphery of necrotic foci was high (46.1–53.9) at 8 and 28 dpi, respectively, and decreased in granulomas found at 28 dpi (6%). Transmission electron microscopy confirmed the presence of apoptotic eosinophils in hepatic lesions at 8 and 28 dpi. At 17 wpi, apoptotic eosinophils were detected in the infiltrate surrounding some enlarged bile ducts containing adult flukes. This is the first report of apoptosis induced by F. hepatica in sheep and the first study reporting apoptosis in eosinophils in hepatic inflammatory infiltrates in vivo. The high number of apoptotic eosinophils in acute necrotic tracts during the migratory and biliary stages of infection suggests that eosinophil apoptosis may play a role in F. hepatica survival during different stages of infection

Apoptosis of peritoneal leucocytes during early stages of Fasciola hepatica infections in sheep

Several immunomodulatory properties have been described in Fasciola hepatica infections. Apoptosis has been shown to be an effective mechanism to avoid the immune response in helminth infections. The aim of the present work was to study apoptosis in peritoneal leucocytes of sheep experimentally infected with F. hepatica during the early stages of infection. Five groups (n = 5) of sheep were used. Groups 2–5 were orally infected with 200 metacercariae (mc) and sacrificed at 1, 3, 9 and 18 days post-infection (dpi), respectively. Group 1 was used as the uninfected control (UC). Apoptosis was detected using three different methods 1) immunocytochemistry (ICC) with a polyclonal antibody anti-active caspase-3; 2) an annexin V flow cytometry assay using the Annexin V-FITC/propidium iodide (PI); and 3) transmission electron microscopy (TEM). The differential leucocyte count revealed that the majority of peritoneal granulocytes were eosinophils, which increased significantly at 9 and 18 dpi with respect to the uninfected controls. The ICC study revealed that the percentage of caspase-3+ apoptotic peritoneal leucocytes increased significantly from 3 dpi onwards with respect to the uninfected controls. The flow cytometry annexin V assay detected a very significant (P < 0.001) increase of apoptotic peritoneal macrophages, lymphocytes and granulocytes, which remained higher than in the UC until 18 dpi. Transmission electron microscopy studies also confirmed the presence of apoptosis in peritoneal eosinophils at 18 dpi. This is the first report of apoptosis induced by F. hepatica in the peritoneal leucocytes of sheep in vivo. The results of this work suggest the importance of apoptosis induction for the survival of the juvenile parasites in the peritoneal migratory stages of infection

A Study on Physical Exercise and General Mobility in People with Cerebral Palsy: Health through Costless Routines

[Abstract] Sedentary behavior (SB) is a common problem that may produce health issues in people with cerebral palsy (CP). When added to a progressive reduction in motor functions over time, SB can lead to higher percentages of body fat, muscle stiffness and associated health issues in this population. Regular physical activity (RPA) may prevent the loss of motor skills and reduce health risks. In this work, we analyzed data collected from 40 people (20 children and teenagers, and 20 adults) who attend two specialist centers in Seville to obtain an up-to-date picture regarding the practice of RPA in people with CP. Roughly 60% of the participants showed mostly mid/severe mobility difficulties, while 38% also had communicative issues. Most of the participants performed light-intensity physical activity (PA) at least once or twice a week and, in the majority of cases, had a neutral or positive attitude to exercising. In the Asociación Sevillana de Parálisis Cerebral (ASPACE) sample test, the higher the International Classification of Functioning, Disability and Health (ICF), the higher the percentage of negative responses to doing exercise. Conversely, in the Centro Específico de Educación Especial Mercedes Sanromá (CEEEMS), people likes PA but slightly higher ratios of positive responses were found at Gross Motor Function Classification System (GMFCS) levels V and II, agreeing with the higher personal engagement of people at those levels. We have also performed a literature review regarding RPA in CP and the use of low-cost equipment. As a conclusion, we found that RPA produces enormous benefits for health and motor functions, whatever its intensity and duration. Costless activities such as walking, running or playing sports; exercises requiring low-cost equipment such as elastic bands, certain smartwatches or video-games; or therapies with animals, among many others, have all demonstrated their suitability for such a purpose.This research was funded by the Spanish Ministry of Science and Innovation, State Plan 2017–2020: Challenges—R&D&I Projects with grant codes PID2019-104323RB-C32 and PID2019-104323RB-C33

Immunohistochemical analysis of knee chondral defect repair after autologous particulated cartilage and platelet-rich plasma treatment in sheep

This study performs an analysis that will enable the evaluation of the quality, durability, and structure of repaired cartilaginous extracellular matrix tissue using an autologous-based particulated autograft cartilage and platelet-rich plasma treatment (PACI + PRP). A single-blind controlled experiment was conducted on 28 sheep to evaluate the efficacy of the PACI + PRP treatment for cartilage defects. Full-thickness 8 mm diameter defects were created in the weight-bearing area of both knees. The right knees received PACI + PRP. The left knees were treated with Ringer’s lactate solution (RLS) or hyaluronic acid (HA) injections. Sheep were euthanized at 9- or 18-months post-surgery. An extensive immunohistochemical analysis was performed to assess collagen types (I, II, III, V, VI, IX, X, XI) and aggrecan positivity. A semiquantitative scoring system provided a detailed evaluation of immunostaining. Collagens and aggrecan scores in the PACI + PRP groups were similar to healthy cartilage. Significant differences were found in collagens associated with matrix maturity (II and V), degradation (IX), structure and mechanics (VI), and hypertrophy (X) between healthy cartilage and RLS- or HA-repaired cartilage. The PACI + PRP treatment advanced the repair cartilage process in chondral defects with mature hyaline cartilage and enhanced the structural and mechanical qualities with better consistent cartilage, less susceptible to degradation and without hypertrophic formation over tim

Utilización del sistema enzima-mediador para la eliminación de los compuestos extraíbles y la lignina en la industria de pasta y papel

Memoria presentada para optar al Diploma de Estudios Avanzados.La madera y otros materiales lignocelulósicos representan la mayor fuente de energía y materia orgánica renovables de la biosfera. La mayor parte de la biomasa del planeta está constituida por materiales lignocelulósicos, por lo que la degradación de estos compuestos representa el proceso biodegradativo más importante en el ciclo del carbono. Los materiales lignocelulósicos incluyendo los productos de origen agrícola y forestal, están constituidos por tres polímeros estructurales- celulosa, hemicelulosa y lignina- y una serie de compuestos de bajo peso molecular solubles en agua o en solventes orgánicos, los denominados extraíbles. La celulosa es el componente mayoritario (alrededor de un 50%), seguida de la lignina (alrededor de un 20%) y de la hemicelulosa (15%) (Fengel y Wegener, 1984). También presentan pequeños contenidos de proteínas y sales minerales (estimadas como ceniza tras la combustión). La madera es el más abundante de los materiales lignocelulósicos y su estructura y composición condiciona su utilización industrial y la posible aplicación de métodos biotecnológicosFinanciado por el proyecto europeo: “White biotechnology for added value products from renewable plant polymers: Design of Taylor-made biocatalysts and new industrial bioprocesses (BIORENEW)” NMP2-CT- 2006-026456 y un contrato I3P de postgrado.Peer reviewe

A new solution to pitch problems in the pulp and paper industry using the laccase-mediator system

Lipophilic extractives in wood and other lignocellulosic materials exert a highly negative impact in pulp and paper manufacturing causing the so-called pitch deposits. As an alternative to physicochemical methods, lipases have been successfully applied to softwood mechanical pulping.1 However, the enzymes and microbial inocula available till present are only effective on some raw materials and processes. Recently, we have shown for the first time the effectiveness of the laccase-mediator system (LMS) in removing pulp lipids regardless the pulping process and raw material used.2,3,4 In these studies, pulp samples from eucalypt kraft pulping, spruce thermomechanical pulping (TMP), and flax soda-anthraquinone (AQ) pulping were treated with laccase in the presence of 1-hydroxybenzotriazole as redox mediator. The gas chromatography/mass spectrometry analyses of the extracts from the enzymatically-treated pulps revealed that most of the lipophilic compounds were efficiently removed by the LMS treatment. Free and conjugated sitosterol, the main responsible for pitch deposits in eucalypt kraft pulp manufacturing, was completely removed. In spruce TMP pulp, LMS degraded most of the resin acids, sterol esters and triglycerides. In the flax soda-AQ pulp, sterols and long-chain fatty alcohols were almost completely removed. Small amounts of oxidation products (including 7-oxositosterol and stigmasta-3,5-dien-7-one mainly derived from free and esterified sitosterol, respectively) were identified. Pulp and papermaking properties of the enzymatically-treated pulps were also evaluated. It is possible to conclude that LMS treatment is an efficient method to remove pitch-causing lipophilic compounds from hardwood, softwood and nonwood paper pulps, at the same time that their lignin content is reduced.Peer reviewe

Exploring the potential of fungal manganese-containing lipoxygenase for pitch control and pulp delignification

The potential of the lipoxygenase from Gaeumannomyces graminis to remove lipophilic extractives from eucalypt and flax pulps was investigated. Pulp treatments were performed with the lipoxygenase both in the presence and absence of linoleic acid, and were followed by a peroxide bleaching stage. The main lipophilic extractives from eucalypt pulp such as conjugated and free sterols decreased up to 40% and 7%, respectively, by the lipoxygenase treatment in the presence of linoleic acid. Different degradation patterns were observed among the lipophilic compounds present in flax pulp, although a high removal of all the extractives classes, including alkanes (21-55%), fatty alcohols (42-61%), and free (16-55%) and glycosylated (45-71%) sterols, was attained in all the lipoxygenase treatments. Reactions of the lipoxygenase with model lipid mixtures were carried out to better understand the degradation patterns observed in pulps. Finally, pulp delignification by the lipoxygenase treatments was also evaluated.This study has been supported by the Spanish projects BIO2007-28719-E and AGL2008-00709 and the EU project BIORE- NEW (NMP2-CT-2006-026456).Peer Reviewe