Digestive aspartic proteases from sábalo (Prochilodus lineatus): Characterization and application for collagen extraction

Acid proteases from sábalo stomach mucosa were recovered using salting-out procedure. This single step produced an enzyme extract purified 1.8-fold over the crude extract with a recovery of 45.1% of its initial proteolytic activity. Sábalo proteases exhibited the highest activity at 45 °C-pH 2.0, showed pH stability between 2.0 and 5.0 and retained more than 70% of its activity after incubation at pH 7.0 for 2 h. Fish extract was unstable at temperatures greater than 45 °C. Its activity was inhibited by pepstatin A but not by PMSF, while EDTA and SDS showed partial inhibitory effects. Presence of CaCl 2 and MgCl 2 increased the proteolytic activity, while increasing concentrations of NaCl strongly decreased it. In addition, compared to the acid extraction method, the use of sábalo enzymatic extract increased 1.7 times the yield of collagen extraction.Fil: Acevedo Gomez, Antonella Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Gomez, Gabriela Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Chamorro, Ester Ramona. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Bustillo, Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Leiva, Laura Cristina Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentin

An alternative method to isolate protease and phospholipase A2 toxins from snake venoms based on partitioning of aqueous two-phase systems

Snake venoms are rich sources of active proteins that have been employed in the diagnosis and treatment of health disorders and antivenom therapy. Developing countries demand fast economical downstream processes for the purification of this biomolecule type without requiring sophisticated equipment. We developed an alternative, simple and easy to scale-up method, able to purify simultaneously protease and phospholipase A2 toxins from Bothrops alternatus venom. It comprises a multiple-step partition procedure with polyethylene-glycol/phosphate aqueous two-phase systems followed by a gel filtration chromatographic step. Two single bands in SDS-polyacrylamide gel electrophoresis and increased proteolytic and phospholipase A2 specific activities evidence the homogeneity of the isolated proteins.Fil: Gomez, Gabriela Noemi. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Acosta, Ofelia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentina. Universidad Nacional del Nordeste; ArgentinaFil: Picó, Guillermo Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; Argentin

Comparative Study of Proteolytic Activities of Venom from Adult and Juvenile Species of Bothrops Alternatus

Because of the technological advance in proteomic essays, ontogenic variations studies about snake venoms have become more relevant in the last years. The differences would be associated with differences in the snake venom protein composition and could have importance in the selection of the samples for production and control of quality of antivenoms. On the other hand, some authors suggested the use of venom from young snakes (yV) in order to purify some of the main components present in major proportions than in venom from adult snakes (aV). In this study, we compared venoms from Bothrops alternatus (yarará grande) from both stages of ontogeny, focusing on the study of the protein bands by SDS-PAGE and also on the study of proteolytic activities (metalloproteinases and serineproteases enzymes) exhibited by these venoms as the main responsible for the physiopathological action exhibited by this animal. Proteolytic activity was assessing by azocasein method, coagulant activity by citrated plasma and then, amidolytic actitivty was evaluated on BApNA. The results showed differences not only in the protein profile, but also in the enzymatic activities from young and adult snake venoms. Young snake venom coagulated the citrated plasma in the half time (6.3 s) the adult snake venom did. The relation yV/aV of proteolytic activity assessed on azocasein was 2 and 23 in the case of amydolitc action. These results highlight the differences in metalloproteinases concentration (responsible of caseinolytic activity) and especially in the serineproteinases composition (tested on BApNA), which would express in low quantities in adult snake venoms. These evidences warn about the necessity to assay the level of protection that the standart antivenom produces against a potential snakebite by young species and, at the same time, look for the development of antivenoms with pools including young and adult organisms.Fil: Garcia Denegri, María Emilia. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Van de Velde, Andrea Carolina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Gomez, Gabriela Noemi. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Nuñez, Sandra Elizabeth. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Otto, Bárbara Vanesa. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; ArgentinaFil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Teibler, Gladys Pamela. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; ArgentinaXX Annual Meeting of the Argentinean Biology Society and VII Meeting of the Uruguayan Society of BiosciencesBuenos AiresArgentinaSociedad Argentina de BiologíaSociedad Uruguaya de Biociencia

A phase II dose-escalation trial of perioperative desmopressin (1-desamino-8-d-arginine vasopressin) in breast cancer patients

Desmopressin (dDAVP) is a well-known peptide analog of the antidiuretic hormone vasopressin, used to prevent excessive bleeding during surgical procedures. dDAVP increases hemostatic mediators, such as the von Willebrand factor (vWF), recently considered a key element in resistance to metastasis. Studies in mouse models and veterinary trials in dogs with locally-advanced mammary tumors demonstrated that high doses of perioperative dDAVP inhibited lymph node and early blood-borne metastasis and significantly prolonged survival. We conducted a phase II dose-escalation trial in patients with breast cancer, administering a lyophilized formulation of dDAVP by intravenous infusion in saline, 30–60 min before and 24 h after surgical resection. Primary endpoints were safety and tolerability, as well as selection of the best dose for cancer surgery. Secondary endpoints included surgical bleeding, plasma levels of vWF, and circulating tumor cells (CTCs) as measured by quantitative PCR of cytokeratin-19 transcripts. Only 2 of a total of 20 patients experienced reversible adverse events, including hyponatremia (grade 4) and hypersensitivity reaction (grade 2). Reactions were adequately managed by slowing the infusion rate. A reduced intraoperative bleeding was noted with increasing doses of dDAVP. Treatment was associated with higher vWF plasma levels and a postoperative drop in CTC counts. At the highest dose level evaluated (2 μg/kg) dDAVP appeared safe when administered in two slow infusions of 1 μg/kg, before and after surgery. Clinical trials to establish the effectiveness of adjunctive perioperative dDAVP therapy are warranted. This trial is registered on www.clinicaltrials.gov (NCT01606072).Fil: Weinberg, Ruth S.. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: Grecco, Marcelo O.. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: Ferro, Gimena S.. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: Seigelshifer, Debora Judith. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: Perroni, Nancy V.. Provincia de Buenos Aires. Ministerio de Salud. Hospital Interzonal de Agudos "Eva Perón"; ArgentinaFil: Terrier, Francisco J.. Instituto Universitario del Hospital Italiano de Buenos Aires; ArgentinaFil: Sánchez Luceros, Analía Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina. Academia Nacional de Medicina de Buenos Aires; ArgentinaFil: Maronna, Esteban. Centro de Educación Médica e Investigaciones Clínicas “Norberto Quirno”; ArgentinaFil: Sánchez Marull, Ricardo. Centro de Educación Médica e Investigaciones Clínicas “Norberto Quirno”; ArgentinaFil: Frahm, Isabel. Centro de Educación Médica e Investigaciones Clínicas “Norberto Quirno”; ArgentinaFil: Guthmann, Marcelo D.. Laboratorio Elea; ArgentinaFil: Di Leo, Daniela. Laboratorio Elea; ArgentinaFil: Spitzer, Eduardo. Laboratorio Elea; ArgentinaFil: Ciccia, Graciela Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Garona, Juan. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; ArgentinaFil: Pifano, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; ArgentinaFil: Torbidoni, Ana Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; ArgentinaFil: Gomez, Daniel Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; ArgentinaFil: Ripoll, Giselle Vanina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; ArgentinaFil: Gomez, Roberto E.. Laboratorio Elea; ArgentinaFil: Demarco, Ignacio A.. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alonso, Daniel Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes; Argentin

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Omecamtiv mecarbil in chronic heart failure with reduced ejection fraction, GALACTIC‐HF: baseline characteristics and comparison with contemporary clinical trials

Aims: The safety and efficacy of the novel selective cardiac myosin activator, omecamtiv mecarbil, in patients with heart failure with reduced ejection fraction (HFrEF) is tested in the Global Approach to Lowering Adverse Cardiac outcomes Through Improving Contractility in Heart Failure (GALACTIC‐HF) trial. Here we describe the baseline characteristics of participants in GALACTIC‐HF and how these compare with other contemporary trials. Methods and Results: Adults with established HFrEF, New York Heart Association functional class (NYHA) ≥ II, EF ≤35%, elevated natriuretic peptides and either current hospitalization for HF or history of hospitalization/ emergency department visit for HF within a year were randomized to either placebo or omecamtiv mecarbil (pharmacokinetic‐guided dosing: 25, 37.5 or 50 mg bid). 8256 patients [male (79%), non‐white (22%), mean age 65 years] were enrolled with a mean EF 27%, ischemic etiology in 54%, NYHA II 53% and III/IV 47%, and median NT‐proBNP 1971 pg/mL. HF therapies at baseline were among the most effectively employed in contemporary HF trials. GALACTIC‐HF randomized patients representative of recent HF registries and trials with substantial numbers of patients also having characteristics understudied in previous trials including more from North America (n = 1386), enrolled as inpatients (n = 2084), systolic blood pressure &lt; 100 mmHg (n = 1127), estimated glomerular filtration rate &lt; 30 mL/min/1.73 m2 (n = 528), and treated with sacubitril‐valsartan at baseline (n = 1594). Conclusions: GALACTIC‐HF enrolled a well‐treated, high‐risk population from both inpatient and outpatient settings, which will provide a definitive evaluation of the efficacy and safety of this novel therapy, as well as informing its potential future implementation

Comunicación interna en el trabajo en equipo de los empleados administrativos de la unidad de gestión educativa local (Ugel) Sur Arequipa, 2019

TesisEl ser humano desde que nace se enfrenta a distinto retos, necesidades, problemas y conflictos, siendo problemas que afijen a nuestro mundo laboral. El colaborar se enfrenta a un conjunto sin sabores, deficientes, problemas, el cual con su inteligencia supera estos factores, por cuanto las personas están para superar dichos retos, es búsqueda de la mejor calidad de vida. Pero aunque no se desee, estos factores son obstáculos que tiene que superar día a día pero el cual hace mella en los objetivos programados para, desarrollo eficiente de la organización, lo cuales a veces van es desmedro de las personas que se ven reflejado en la frustración y desanimo en ellos. Uno de los retos que tiene cada organización es, las Tecnologías de la Información en la Comunicación (TIC), innovación, el cual hará que las organización sean competitivas y la comunicación es parte fundamental en todo centro laboral por cuanto se realizan diversos proceso de comunicación tanto verbal como no verbal estando las personas se encuentran interactuando constantemente y para ello se requiere de una comunicación eficaz, oportuna. Al hablar de las organizaciones podemos pensar en unidades sociales, compuestas de dos o más personas que interactúan entre sí y que necesitan de la comunicación para poder relacionar. Es por eso que la comunicación dentro de las organizaciones, incluye mucho más que mensajes verbales y no verbales, es hablar de relaciones humanas, de interacción, de convivencia. Dentro de las organizaciones la comunicación interna, en diversas organización no se le ha otorgado ls importancia que esta se merece, si se le diera mayor importancia se podrían mejorarían los procesos comunicacionales y sus diversas herramientas que se encuentran interrelacionadas con ella como el trabajo en equipo la cual reúne a un grupo de colaboradores con diversas capacidades, ideas, destrezas, habilidades con el objetivo de hacerlo muchas más rápido y obtener resultados óptimos el cual generara crear sinergia, empoderamiento, promover estructuras más flexibles y con menor jerarquía, fomentar el trabajo multidisciplinario, la responsabilidad y la capacidad de respuesta al cambio, promover sentido de logro y equidad y la amistad y en forma transversal se encuentra la comunicación y estos aspectos incidirán en la mejora de la calidad de vida laboral y en consecuencia se mejorara el desarrollo laboral. El estar bien informados constituye una fuente de energía importante para la organización, para poder tomar decisiones respecto a los aspectos importantes del entorno de esta. Para el desarrollo de la investigación se eligió la investigación cuantitativa, el cual será de tipo no experimental, transeccional de tipo correlacional. El presente trabajo tiene como propósito conocer cómo afecta la Comunicación interna en los equipos de trabajo en los empleados administrativos de la UGEL SUR AREQUIPA 2019

Aqueous two-phase systems: A simple methodology to obtain mixtures enriched in main toxins of Bothrops alternatus venom

Phospholipase A2 (PLA2) and protease (P) are enzymes responsible of myotoxic, edematogenic and hemostasis disorder effects observed in the envenomation byBothrops alternatus pitviper. Their partitioning coefficient (Kp) in different polyethyleneglycol/potassium phosphate aqueous two-phase systems (ATPSs) was determined in order to both achieve a better understanding of the partitioning mechanism and define optimal conditions for toxin isolation. Polyethyleneglycols (PEGs) of molecular weights 1000; 3350; 6000 and 8000; different temperatures (5, 20 and 37 °C) and phase volume ratios of 0.5; 1 and 2 were assayed. PLA2 partitioned preferentially to the top phase while P mainly distributed to the bottom phase. Either entropically- or enthalpically-driven mechanisms were involved in each case (PLA2 and P). The aqueous two-phase system formed by PEG of MW 3350 (12.20% wt/wt) and KPi pH 7.0 (11.82% wt/wt) with a volume ratio of one and a load of 1.25 mg of venom/g of system showed to be the most efficient to recover both enzymes. It allowed obtaining the 72% of PLA2 in the top phase with a purification factor of 2 and the 82% of P at the bottom phase simultaneously. A further adsorption batch step with DEAE-cellulose was used to remove satisfactorily the PEG from the top phase and recover the active PLA2. The proposed methodology is simple, inexpensive, and only requires professionals trained in handling basic laboratory equipment. It could be easily adoptable by developing countries in which the snakebite accidents cause considerable morbidity and mortality.Fil: Gomez, Gabriela Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Leiva, Laura Cristina Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; Argentin

Pacu viscera extract obtained by aqueous two-phase system: potential application in recycling waste

Pacú (Piaractus mesopotamicus) production represents 40% of northeast fish yield. It is of interest to valorize the current disposal of fish processing through the use of viscera waste, source of enzymes such as trypsin. Aqueous two-phase systems (ATPS) have been successfully used for separation and purification of macromolecules because exhibit multiple advantages: good resolution, high yield, low cost and proteins retain their biological activity. The goal of this work was to analyze the partition of pacú pyloric caeca extract in aqueous two-phase PEG-citrate systems. Preparation of crude alkaline extract was made by mechanical and sonic digestion of pyloric caeca. A two-phase system formed by polyethyleneglycol (PEG) 3350 - citrate, pH 8.4, was used and the partition of alkaline proteins of the extract was assayed. Trypsin activity was determined with the substrate a-Nbenzoyl-DL-arginine-p-nitroanilide (BAPNA) and protein content was estimated by the Warburg and Christian method. Partition coefficients (Kp) were calculated by the ratio of enzyme trypsin activity or total protein between phases. The experiments revealed that partition coefficient of trypsin was greater than unity (KpTrip: 2,2), while partition coefficient of total proteins was smaller than one (KpTP: 0,33). This result indicates a tendency for trypsin-like enzymes to concentrate in the top phase (PEG-enriched phase), while proteins were partitioned to the salt-enriched phase (bottom phase). Thus, a high separating capability was verified in the assayed system. On the other hand, X ray plates were treated with the top phase achieving a complete removed of the gelatine layer which covers it, with no interference of PEG. These studies demonstrate the applicability of a simple procedure to obtain a phase enriched with enzymes from pacú viscera extracts and their use in recycling of radiographic plates.Fil: Gomez, Gabriela Noemi. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Acevedo Gomez, Antonella Valeria. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Modelado e Innovación Tecnológica. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Modelado e Innovación Tecnológica; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaFil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; ArgentinaXX Annual Meeting of the Argentinean Biology Society (SAB) . VII Meeting of the Uruguayan Society of Biosciences (SUB). Second Biology Meeting Of The River PlateBuenos AiresArgentinaSociedad Argentina de BiologíaSociedad Uruguaya de Biociencia

Aqueous micellar two phase systems as novel tools to recover pepsin like surubí proteases: effect of surfactants on enzyme activity

Surubí is farmed in the northeast of Argentina. It is of interest to valorize the current disposal of fish processing through the use of viscera waste, source of enzymes such as Pepsin. These enzymes have been recovered mainly using conventional methods (e.g.salting out, chromatography), however the use of liquid-liquid extraction using surfactants have not been fully explored. Aqueous micellar two-phase systems (AMTPS) are an extractive method based on the ability of some surfactants to form two immiscible aqueous phases, a rich and a poor micelle phases to recover the product of interest. Thus, the objective of this work was to firstly evaluate the stability of surubí crude extracts under different concentrations of Genapol (GX080) and Tergitol (Tg7) to estimate the feasibility of their use as potential micellar extractants of surubí pepsin. Enzymatic extracts (ERPi) were recovered from stomachs homogenates using salting-out procedure and pepsin activity was estimated by acid hemoglobin method. ERPi were first incubated with various concentrations of surfactants (1, 3 and 5% GX080/Tg7) in 100mM NaCit pH 5 for different times (0, 1, 2 and 3h) and then enzymatic activity was measured. Data represent the mean ± standard deviation (SD) of at least three replications. Statistical significance was tested by one-way ANOVA and Tukey (HSD) (p<0.05) Results showed that any surfactant concentration tested affected ERPi enzymatic activity moderately and this effect was dose dependent. ERPi retained 92, 65 and 60% of its initial activity when were incubated with 1, 3 and 5% of Tg7. After incubating with GX080 (1, 3 and 5%), the ERPi exhibited 80, 70 and 60% of its initial activity, respectively. The time variable in these assays had not significant influence on enzymatic activity decrease. Pepsin, a hydrophilic protein, is expected to be preferentially distributed into the aqueous phase of AMTPS.Considering that the surfactant concentration at this phase is always below 1%, the obtained results suggest that AMTPS formed with either GX080 or Tg7 could be viable tools in the primary recovery of pepsin-like proteases from fishing waste.Fil: Acevedo Gomez, Antonella Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Escobar, Guilermo. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; ArgentinaFil: Gomez, Gabriela Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Leiva, Laura Cristina Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Bustillo, Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Químicos Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Procesos Biotecnológicos y Químicos Rosario; ArgentinaReunión Anual de Sociedades de BiocienciaMar del PlataArgentinaSociedad Argentina de Farmacologia ExperimentalSociedad Argentina de Investigación ClínicaSociedad Argentina de BiologíaSociedad Argentina de ProtozoologíaAsociación Argentina de NanomedicinasAsociación Argentina de Ciencia y Tecnología de Animales de Laboratori