Neutralizing capacity of antisera obtained by immunization with Bothrops alternatus venom blocked in its metalloproteinases

Specific treatment for snake bite accidents with antivenoms, obtained from immunized animals, is the only recommended therapy. Bothropic intoxication is characterized by proteolytic, coagulant and hemorrhagic effects that induce local tissue damage and systemic alterations that could lead to death. Snake venom metalloproteinases, zinc- dependent, play a relevant role in the pathogenesis of intoxication. They provoke a disruption of the hemostatic system, restricting their use as immunogen in high doses. In previous work, it was demonstrated that is it possible to immunize mice with high doses of Bothrops alternatus venom (BaV) neutralized with disodium salt of ethylenediaminetetraacetic acid (Na2EDTA) avoid of hemorrhagic lesions and then animals reported higher antibodies titer. The present study was performed to test the neutralizing ability of sera obtained from animals treated whit venom-Na2EDTA, and evaluate the effect of this immunogen on pulmonary parenchyma. Groups of 5 Balb/c mice were immunized subcutaneously with BaV, or BaV/Na2EDTA emulsified with Freund?s Adjuvant (complete first and incomplete-booster). On day 50 serums were collected for neutralization assays: proteolytic activity on azocasein, phospholipase activity on erythrocytes and thrombine-like activity on citrated plasma. Animals were sacrificed and lungs removed for histological analysis (hematoxylin-eosin). The results showed that the capacity of neutralize each of one of the three enzyme activities assayed was significantly higher (p <0.05) by the serum obtained from animals immunized with BaV/Na2EDTA. Histological analysis showed that the pulmonary parenchyma from immunized mice with BaV was significantly affected (pneumonitis, p <0.05), in respect to those were BaV/Na2EDTA treated. Results demonstrated that the BaV/Na2EDTA immunogen has a lower organic impact with respect to BaV and, at the same time, providing a serum with high neutralizing capacity.Fil: López, Gisela Lumila. 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; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; ArgentinaFil: Hernandez, David Roque. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias. Instituto de Ictiología del Nordeste; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; 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. 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: Fusco, Luciano Sebastian. 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: 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; ArgentinaLXIV Reunión Anual de la Sociedad Argentina de Investigación Clínica; LI Reunión Anual de la Asociación Argentina de Farmacología Experimental; XXI Reunión Anual de la Sociedad Argentina de Biología; XXXI Reunión Anual de la Sociedad Argentina de Protozoología; IX Reunión Anual de ka Asociación Argentina de Nanomedicinas y VI Reunión Científica Regional de la Asociación Argentina de Ciencia y Tecnología de Animales de LaboratorioMar del PlataArgentinaSociedad Argentina de Investigación ClínicaAsociación Argentina de Farmacología ExperimentalSociedad Argentina de BiologíaAsociación Argentina de NanomedicinasAsociación Argentina de Ciencia y Tecnología de Animales de LaboratorioSociedad Argentina de Protozoologí

Role of the orexin system on the hypothalamus-pituitary-thyroid axis

Hypocretin/orexin (ORX) are two hypothalamic neuropeptides discovered in 1998. Since their discovery, they have been one of the most studied neuropeptide systems because of their projecting fields innervating various brain areas. The orexinergic system is tied to sleep-wakefulness cycle, and narcolepsy is a consequence of their system hypofunction. Orexinergic system is also involved in many other autonomic functions such as feeding, thermoregulation, cardiovascular and neuroendocrine regulation. The main aim of this mini review article is to investigate the relationship between ORX and thyroid system regulation. Although knowledge about the ORX system is evolving, its putative effects on hypothalamic-pituitary-thyroid (HPT) axis still appear unclear. We analyzed some studies about ORX control of HPT axis to know better the relationship between them. The studies that were analyzed suggest Hypocretin/ORX to modulate the thyroid regulation, but the nature (excitatory or inhibitory) of this possible interaction remains actually unclear and needs to be confirmed

Osteopontin: Relation between Adipose Tissue and Bone Homeostasis

Osteopontin (OPN) is a multifunctional protein mainly associated with bone metabolism and remodeling. Besides its physiological functions, OPN is implicated in the pathogenesis of a variety of disease states, such as obesity and osteoporosis. Importantly, during the last decades obesity and osteoporosis have become among the main threats to health worldwide. Because OPN is a protein principally expressed in cells with multifaceted effects on bone morphogenesis and remodeling and because it seems to be one of the most overexpressed genes in the adipose tissue of the obese contributing to osteoporosis, this mini review will highlight recent insights about relation between adipose tissue and bone homeostasis

Antifungal Activity of Decyl Gallate against Several Species of Pathogenic Fungi

This work aims to demonstrate that the gallic acid structure modification to the decyl gallate (G14) compound contributed to increase the antifungal activity against several species of pathogenic fungi, mainly, Candida spp., Cryptococcus spp., Paracoccidioides spp., and Histoplasma capsulatum, according to standardized microdilution method described by Clinical Laboratory Standard Institute (CLSI) documents. Moreover this compound has a particularly good selectivity index value, which makes it an excellent candidate for broad-spectrum antifungal prototype and encourages the continuation of subsequent studies for the discovery of its mechanism of action

Lopinavir/Ritonavir and Darunavir/Cobicistat in Hospitalized COVID-19 Patients: Findings From the Multicenter Italian CORIST Study

Background: Protease inhibitors have been considered as possible therapeutic agents for COVID-19 patients. Objectives: To describe the association between lopinavir/ritonavir (LPV/r) or darunavir/cobicistat (DRV/c) use and in-hospital mortality in COVID-19 patients. Study Design: Multicenter observational study of COVID-19 patients admitted in 33 Italian hospitals. Medications, preexisting conditions, clinical measures, and outcomes were extracted from medical records. Patients were retrospectively divided in three groups, according to use of LPV/r, DRV/c or none of them. Primary outcome in a time-to event analysis was death. We used Cox proportional-hazards models with inverse probability of treatment weighting by multinomial propensity scores. Results: Out of 3,451 patients, 33.3% LPV/r and 13.9% received DRV/c. Patients receiving LPV/r or DRV/c were more likely younger, men, had higher C-reactive protein levels while less likely had hypertension, cardiovascular, pulmonary or kidney disease. After adjustment for propensity scores, LPV/r use was not associated with mortality (HR = 0.94, 95% CI 0.78 to 1.13), whereas treatment with DRV/c was associated with a higher death risk (HR = 1.89, 1.53 to 2.34, E-value = 2.43). This increased risk was more marked in women, in elderly, in patients with higher severity of COVID-19 and in patients receiving other COVID-19 drugs. Conclusions: In a large cohort of Italian patients hospitalized for COVID-19 in a real-life setting, the use of LPV/r treatment did not change death rate, while DRV/c was associated with increased mortality. Within the limits of an observational study, these data do not support the use of LPV/r or DRV/c in COVID-19 patients

ATLANTIC-CAMTRAPS: a dataset of medium and large terrestrial mammal communities in the Atlantic Forest of South America

Our understanding of mammal ecology has always been hindered by the difficulties of observing species in closed tropical forests. Camera trapping has become a major advance for monitoring terrestrial mammals in biodiversity rich ecosystems. Here we compiled one of the largest datasets of inventories of terrestrial mammal communities for the Neotropical region based on camera trapping studies. The dataset comprises 170 surveys of medium to large terrestrial mammals using camera traps conducted in 144 areas by 74 studies, covering six vegetation types of tropical and subtropical Atlantic Forest of South America (Brazil and Argentina), and present data on species composition and richness. The complete dataset comprises 53,438 independent records of 83 species of mammals, includes 10 species of marsupials, 15 rodents, 20 carnivores, eight ungulates and six armadillos. Species richness averaged 13 species (±6.07 SD) per site. Only six species occurred in more than 50% of the sites: the domestic dog Canis familiaris, crab-eating fox Cerdocyon thous, tayra Eira barbara, south American coati Nasua nasua, crab-eating raccoon Procyon cancrivorus and the nine-banded armadillo Dasypus novemcinctus. The information contained in this dataset can be used to understand macroecological patterns of biodiversity, community, and population structure, but also to evaluate the ecological consequences of fragmentation, defaunation, and trophic interactions. © 2017 by the Ecological Society of Americ

Selenium in the thyroid: physiology and pathology

Selenium (Se) is a trace element that plays a critical role in several processes for human health. The thyroid is the organ with the highest Se content per gram of tissue; in thyroid follicular cells, Se acts as antioxidant by contrasting the production of the reactive oxygen species that are generated during thyroid hormones biosynthesis. In addition, Se is part of the active site of the deoidinases, the enzymes responsible for thyroid hormones activation and inactivation. Herein, the effects of Se supplementation in patients with thyroid related disorders have been reviewed on the basis of the studies published on this issue

Bothrops alternatus venom pretrated with chelate agent for antiserum production

Snakebite envenoming is a major public health problem in tropical countries. The only specific therapies currently available for the treatment are antivenoms, which consist of polyclonal immunoglobulins purified from sera/plasma of horses or sheep immunized with doses sublethal of snake venom(s). Bothrops alternatus (BaV) venom induces a prominent tissue local damage, especially, hemorrhage, muscle damage and inflammation. These complex pathological phenomena are due to the concomitant action of metalloproteinases (SVMPs) in venoms and others edema-inducing components. In order to obtain a high titer of antibodies and reduced local damage in the animal, an alternative immunization protocol was proposed where the SVMPs action was blocked by using a chelate agent. For this proposal, the B. alternatus venom (10 mg/mL) was blocked by EDTA-Na2 (10 mM, BaV/ EDTA-Na2) and used as antigen. Previously to the inoculation, the excess of chelate was removed by passing the mixture on Sephadex G-25 column (venom without inhibitor was subjected to the same process) and the effective neutralization of SVMPs using azocasein as substrate was determined. Group of 5 Balb/c mice were immunized subcutaneously on days 0, 15 and 30 with BaV(15-30-45μg) or BaV/ EDTA-Na2 (45-90-135 μg) emulsified with Freund?s Adyuvant (complete first and incomplete-booster). Blood samples were collected by the animals tail tip on days 14, 29 and 41 of protocol immunization and it was destined to ELISA´s test. The results showed that the immunized animal with BaV/ EDTA-Na2had a higher titer (5.1x104) than those treated to BaV (1.3x104). Macroscopic analysis at the inoculation site of mice injected with Freund?s adjuvant showed local damage (with non-infectious abscesses) and hypertrophy of inguinal lymph nodes. Our results show that BaV/ EDTA-Na2 formulation, where the SVMPs are blocked, produced a higher humoral response compared with the produced by BaV. These preliminary results demonstrated the potential use of blocking the toxins with chelate to produce antivenom with less damage in the animals.Fil: López, Gisela Lumila. 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; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; 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: 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. 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: Fusco, Luciano Sebastian. 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; ArgentinaXX Annual Meeting of the Argentinean Biology Society and VII Meeting of the Uruguayan Society of BiosciencesBuenos AiresArgentinaSociedad Argentina de BiologíaSociedad Uruguaya de Biociencia

Use of venom blocked with chelating agent for anti-bothropic serum production

Snakes belonging to genus Bothrops are responsible for more than 85% of the bites occurring in South America. Bothrops alternatus is a pitviper widespread in this area and it is one of the most important species associated to snakebites not only in Argentina but also Brazil. Antivenoms are the only specific treatment for envenoming by snakebites. They are made by animals (e.g. horses or sheep) immunizations with sublethal doses of venom. Snake Venom Metalloproteinases (SVMPs) play an important role in envenomation, causing relevant local effects such as hemorrhage, edema and myotoxicity as well as systemic bleeding. They represent around ̴43.1% of the protein composition from B. alternatus venom and causing lesions during the immunization of animals. In this work, an alternative immunization protocol was developed in mice where SVMPs activity was previously blocked by Na2EDTA as chelating agent. For this proposal, the B. alternatus venom (BaV, 10 mg/mL) was treated with 50 mM Na2EDTA (1 h, 37°C) and excess chelator was removed by Sephadex G-25 chromatography. Proteolytic activity was assayed to control the block processing. Groups of 5 BALB/c mice were immunized s.v. on 0-15-30 days with BaV (15-30-45μg) and BaV/Na2EDTA (45-90-135 μg). Blood samples were collected on days 14-29-41 for antibody analysis. Sera from BaV/Na2EDTA protocol have a titer (5.1x104) higher than those treated with BaV (1.3x104). The neutralizing ability of both antivenoms was tested against proteolytic, coagulant and PLA2 activity, resulting that it was significantly higher (p <0.05) for sera from mice bowing to blocked venom. Histological analysis of mice lungs showed that pulmonary parenchyma from BaV immunized mice was significantly affected in relation to those BaV/Na2EDTA treated. In conclusion, BaV/Na2EDTA showed to be an appropriate immunogen, not only proved that serum has a high neutralizing capacity but also has a lower organic impact in animals to antivenom production.Fil: López, Gisela Lumila. 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: Van de Velde, Andrea Carolina. 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: Hernandez, David Roque. Universidad Nacional del Nordeste. Facultad de Ciencias Veterinarias. Instituto de Ictiología del Nordeste; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; 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: Fusco, Luciano Sebastian. 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; Argentina7th International Toxinology MeetingOxfordInglaterraLibPubMedi