Avaliação in vivo da pirogenicidade de bioprodutos fúngicos com potencial prebiótico

Introduction: natural products with pharmacological activity require preclinical evaluations to justify their uses scientifically. The pyrogen assay is a safety pharmacology test performed to determine the presence of endotoxins and it is a valuable method to demonstrate the bio-derivative products safety and their prebiotic potential in the field of immunonutrition. Objective: to evaluate the pyrogenicity of fungal bioproducts from Pleurotus ostreatus (aqueous extracts from mycelium and fruiting bodies) and a biopreparation from Kluyveromyces marxianus yeast, using a pyrogen assay in New Zealand rabbits. Method: concentrations of 1.0 and 10.0 mg/mL of each sample were tested intravenously at doses of 0.5 and 5.0 mg/kg body weight. The experimental design complied with good laboratory practices as established by the International Council for Laboratory Animal Science and was carried out according to the standard work procedures of the Centro de Toxicología y Biomedicina, Santiago de Cuba. Results: Pleurotus ostreatus extracts and the yeast biopreparation (0.5 mg/kg) showed no signs of pyrogenicity. In the biopreparation results (5.0 mg/kg), temperature values fall in the uncertainty range according to the United States Pharmacopoeia (USP), and therefore it was suggested to repeat the study. Conclusions: Pleurotus ostreatus extracts and Kluyveromyces marxianus biopreparation (0.5 mg/kg) did not induce a significant temperature increase in the animals, thereby suggesting that there are no endotoxin levels in such bioproducts that could cause pyrogenicity.Introducción: los productos naturales con actividad farmacológica requieren de evaluaciones preclínicas que justifiquen su empleo sobre una base científica. El ensayo de pirógenos es una prueba dentro de la Farmacología de Seguridad que se realiza para determinar la presencia de endotoxinas y constituye un método valioso, para demostrar la seguridad de bioderivados con potencial prebiótico en el campo de la inmunonutrición. Objetivo: evaluar la pirogenicidad de bioproductos fúngicos de Pleurotus ostreatus (extractos acuosos del micelio y cuerpos fructíferos) y un biopreparado de levadura Kluyveromyces marxianus, empleando el ensayo de pirógenos en conejos Nueva Zelanda. Método: se ensayaron concentraciones de 1,0 y 10,0 mg/mL de cada muestra por vía endovenosa en dosis de 0,5 y 5,0 mg/kg de peso. El diseño experimental cumplió las buenas prácticas de laboratorio según lo establecido por el International Council for Laboratory Animals Science y se realizó de acuerdo a los procedimientos normalizados de trabajo del Centro de Toxicología y Biomedicina, Santiago de Cuba. Resultados: los extractos de Pleurotus ostreatus y el biopreparado de levadura (0,5 mg/kg) no mostraron signos de pirogenicidad. En los resultados del biopreparado (5,0 mg/kg), los valores de temperatura caen en un rango de incertidumbre, según la Farmacopea de Estados Unidos (USP) y se sugirió repetir el estudio.Conclusiones: los extractos de Pleurotus ostreatus y el biopreparado de Kluyveromyces marxianus (0,5 mg/kg) no indujeron un aumento de temperatura significativo en los animales, lo cual sugiere que en estos bioproductos no existen niveles de endotoxinas que puedan provocar pirogenicidad.Introdução: produtos naturais com atividade farmacológica requerem avaliações pré-clínicas que justifiquem seu uso em bases científicas. O ensaio de pirogênio é um teste dentro da Farmacologia de Segurança que é realizado para determinar a presença de endotoxinas e é um método valioso para demonstrar a segurança de bioderivados com potencial prebiótico no campo da imunonutrição.Objetivo: avaliar a pirogenicidade de bioprodutos fúngicos de Pleurotus ostreatus (extratos aquosos do micélio e corpos de frutificação) e de uma biopreparação da levedura Kluyveromyces marxianus, utilizando o ensaio pirogênico de coelho da Nova Zelândia. Método: concentrações de 1,0 e 10,0 mg/mL de cada amostra foram testadas por via intravenosa nas doses de 0,5 e 5,0 mg/kg de peso. O desenho experimental obedeceu às boas práticas laboratoriais estabelecidas pelo Conselho Internacional para a Ciência dos Animais de Laboratório e foi realizado de acordo com os procedimentos de trabalho padrão do Centro de Toxicologia e Biomedicina, Santiago de Cuba. Resultados: os extratos de Pleurotus ostreatus e a biopreparação de leveduras (0,5 mg/kg) não apresentaram sinais de pirogenicidade. Nos resultados da biopreparação (5,0 mg/kg), os valores de temperatura estão dentro de uma faixa de incerteza, segundo a Farmacopeia dos Estados Unidos (USP) e foi sugerido repetir o estudo. Conclusões: os extratos de Pleurotus ostreatus e a biopreparação de Kluyveromyces marxianus (0,5 mg/kg) não induziram um aumento significativo da temperatura nos animais, o que sugere que não há níveis de endotoxinas nesses bioprodutos que possam causar pirogenicidade

Obtención y adaptación de un hibridoma productor de anticuerpos anti-C3 a medio libre de suero

El suero de Coombs o reactivo antiglobulínico humano poliespecífico permite visualizar la presencia de anticuerpos anti-eritrocitarios incompletos o componentes del factor C3 del sistema del complemento humano adheridos a sus membranas. Es de gran importancia clínica para estudiar patologías como la enfermedad hemolítica del recién nacido, enfermedades hemolíticas autoinmunes, entre otras. Actualmente existe la necesidad de cubrir la demanda hospitalaria a través de la producción nacional. El hibridoma secretor de anticuerpos monoclonales anti-C3 generado para la producción del reactivo de Coombs debe pasar por un proceso de adaptación a medio libre de suero y proteínas debido a las desventajas que puede traer el medio que se usa tradicionalmente para su cultivo. El presente tema de investigación propone demostrar que los medios libres de suero y proteínas seleccionados son adecuados para el cultivo del hibridoma. Se realizó un estudio experimental en el Laboratorio de anticuerpos y biomodelos experimentales, de Santiago de Cuba desde abril a noviembre de 2022. Se obtuvieron linfocitos B anti-C3 a partir de la sensibilización de ratones BALB/c. Se generó un hibridoma secretor de anticuerpos anti-C3 y se evaluó su adaptación en las formulaciones preparadas a partir de los medios MB02, MB03 y MB04, todas de producción nacional por lo que proporcionan ventajas económicas. El anticuerpo monoclonal obtenido podrá ser utilizado para el desarrollo de un reactivo antiglobulínico humano poliespecífico que permitirá satisfacer la demanda hospitalaria nacional, garantizando la calidad de la atención en la salud pública y la sustitución de importaciones

Protein hydrolysates from the alga Chlorella vulgaris 87/1 with potentialities in immunonutrition

ABSTRACT Chlorella vulgaris (Chlorophyta, Chlorophyceae) has received a particular attention in the programmes of microalgae utilisation in biotechnology. Enzymatic hydrolysis of cell proteins represents a very promising method to increase protein digestibility and thus, for obtaining hydrolysates with improved nutritional and functional properties. However, this technology has been little approached and the biological evaluation of hydrolysates has had a strictly nutritional nature. The design of hydrolysis conditions that combined for the first time, the use of C.vulgaris 87/1 treated with ethanol and pancreatin at pH values of 7.5-8.0, led to a product with a degree of hydrolysis of 20-22% and yields of 50-55%, characterised by a high digestibility (97.2%) and nitrogen solubility over a wide pH range (2.0-10.0). Hydrolysis curves were fitted to an exponential model, common to many food proteins. The bulk of the product dry matter consists of soluble peptides and free amino acids (47.7%) with three main peptides of molecular masses between 2 and 5 kDa. The oral administration of Chlorella hydrolysate (500 mg/kg) to undernourished Balb/c mice provided benefits in terms of liver protein metabolism and the induction of anabolic processes in gut mucosa. The hydrolysate also enhanced the immunological recovery, as judged by the stimulation of haemopoiesis, monocytemacrophage system activation, as well as humoral and cell mediated immune functions, like T-dependent antibody response and the reconstitution of delayed-type hypersensitivity (DTH) response. These results represent the first findings in the world concerning the immunomodulating effects of a microalgae protein hydrolysate

La Bioinformática para el perfeccionamiento Curricular de la carrera Licenciatura en Biología

Bioinformatics has been a rapidly evolving interdisciplinary science in the last years due tothe great availability of biological information. The numerous applications of this science inBiotechnology, Pharmacology, Biology and Medicine make it an attractive research area,and currently there is a growing demand of trained personnel in this field. However, thescarcity of qualified personnel and the limited incorporation of this content as educationalresource in our university are evident. The present work is referred to an educationalexperience in the Bachelor in Biology major with the incorporation of the optional subjectIntroduction to the Bioinformatics in the fourth year. This subject has a strong practicalcomponent for the formation of skills in the students and it is taught by a team make up ofteachers. The general topic chosen for the practical activities was the analysis of theHemoglobin, topic that influences positively in the motivation of students. The resultsshow that this experience improves the quality of the teaching and learning process,based on the integration of different knowledge areas promotes an autonomous learningand dedication of the students.La Bioinformática es una ciencia ha tenido un desarrollo acelerado debido a la gran disponibilidad de información biológica. Sus aplicaciones en la Biotecnología, Farmacología, Biología y Medicina hacen de esta, un área atractiva de investigación. Sin embargo, la carencia de personal calificado y la limitada incorporación de este contenido son evidentes. Se expone experiencia en la carrera Licenciatura en Biología en la asignatura optativa Introducción a la Bioinformática en el cuarto año. El tema escogido para las clases prácticas fue el análisis de la Hemoglobina. Los resultados muestran mejora en la calidad del proceso de enseñanza aprendizaje de esta asignatura, a partir de la integración de saberes de diferentes disciplinas promueve un aprendizaje autónomo y la calidad del estudio de los estudiantes

Hypnitz: El angelito del abismo

Joc casual basat en minijocs en el que el protagonista, Hypnitz, fill de Satanàs, en l’intent de demostrar que té maldat i que mereix ser el successor al tron, puja al món dels humans per a sembrar la por i demostrar la seva vàlua com a successor al tron de d’infern

Hypnitz: El angelito del abismo

Joc casual basat en minijocs en el que el protagonista, Hypnitz, fill de Satanàs, en l’intent de demostrar que té maldat i que mereix ser el successor al tron, puja al món dels humans per a sembrar la por i demostrar la seva vàlua com a successor al tron de d’infern

Explorando nuevas facetas en el papel de los alimentos funcionales/ nutracéuticos en la salud humana: una mirada a las setas comestibles-medicinales

International audienc

Mycelia from Pleurotus sp. (oyster mushroom): a new wave of antimicrobials, anticancer and antioxidant bio-ingredients

International audienceBackground-There has been an upsurge of interest in mushrooms, such as Pleurotus species, as an important source of bioactive compounds. Mycelia-submerged culture represents a promising approach to search new safe and healthy myco-products with standardized quality in addition to mushroom fruiting bodies. Methods-The study examined the in vitro antimicrobial, antitumor and antioxidant activities of a hot-water extract from Pleurotus sp. mycelium. The antimicrobial activity was screened through the activation of the microbial autolytic system of four bacteria and four yeast strains. The anti-proliferative effects on NB4 human leukemia cells were measured by flow-cytometry analyses. The antioxidant activity was investigated by the scavenging of DPPH and ABTS radicals, the reducing power and the inhibition of lipid peroxidation. Results and discussion-The extract activated the microbial autolytic system of eight strains: seven autolyzing strains with intensity values (Is) ranging from 2.7% in Candida sp. to 36.1% in Saccharomyces cerevisiae. Thus, the microbial autolytic system of the strains tested (including Gram-positive and Gram-negative bacteria as well as yeasts) could be activated in vitro by mycelial extract. Pleurotus extract reduced the viability of NB4 leukemia cells, particularly at the concentration of 200 μg/mL to 82% compared to control cells, and induced apoptosis demonstrated by an increase in annexin V-FITC+ cells (25% at 200 μg/mL). At 10 mg/mL, the extract showed the most potent scavenging effects for DPPH and ABTS radicals (96% and 55%, respectively) and the inhibition of lipid peroxidation (52%). The mushroom extract at 5 mg/mL manifested reducing power of 1.105. Although carbohydrates (76.8%, w/w) appear to be the most important bioactive compounds, secondary metabolites, like phenolics, would also contribute to the antioxidant, antimicrobial and anti-proliferative activities. Conclusions-The hot-water extract obtained from Pleurotus mycelium, in light of its in vitro antimicrobial, antitumor and antioxidant effects could be considered a good candidate for developing nutraceuticals and for designing innovative myco-therapeutics and phytocosmetics applications

Nocardiopsis dassonvillei subsp. crassaminis subsp. nov., isolated from freshwater sediment, and reappraisal of Nocardiopsis alborubida Grund and Kroppenstedt 1990 emend. Nouioui et al. 2018

An actinomycete, strain D1(T), was isolated from a freshwater sediment sample collected from the San Pablo river in the La Risuena community, Santiago de Cuba province, Cuba. The strain was identified as a member of the genus Nocardiopsis by means of a polyphasic taxonomic study. It produced a light yellow non-fragmented substrate mycelium, a white well-developed aerial mycelium and straight to flexuous hyphae. No specific spore chains were observed. Strain D1(T) contained meso-diaminopimelic acid, no diagnostic sugars, and MK-10(H-2), MK-10(H-4), MK-10 and MK-10(H-6) as predominant menaquinones, but not phosphatidylcholine as diagnostic polar lipid of the genus Nocardiopsis. The predominant fatty acids were iso-C-16:0, 10-methyl-C-18:0 and anteiso-C-17:0. Strain D1(T) showed the highest degree of 16S rRNA gene sequence similarity to Nocardiopsis synnematoformans DSM 44143(T) (99.8 %), Nocardiopsis dassonvillei subsp. albirubida NBRC 13392(T) (99.8 %) and Nocardiopsis dassonvillei subsp. dassonvillei DSM 43111(T) (99.6 %). A genomic OrthoANIu value between D1(T) and N. dassonvillei subsp. dassonvillei DSM 43111(T) of 97.63% and a dDDH value of 78.9% indicated that strain D1(T) should be classified in N. dassonvillei. However, phenotypic characteristics distinguished strain D1(T) from its nearest neighbour taxon. On basis of these results we propose to classify strain D1(T) (=LMG 30468(T)=CECT 30033(T)) as a representative of a novel subspecies of the genus Nocardiopsis, for which the name Nocardiopsis dassonvillei subsp. crassaminis subsp. nov. is proposed. In addition, the genomic distance between N. dassonvillei subsp. albirubida NBRC 13392(T) and N. dassonvillei subsp. dassonvillei DSM 43111(T) as determined through OrthoANIu (93.64 %) and dDDH (53.40%), along with considerable phenotypic and chemotaxonomic differences reported in earlier studies, indicated that the classification of this taxon as Nocardiopsis alborubida Grund and Kroppenstedt 1990 is to be preferred over its classification as N. dassonvillei subsp. albirubida Evtushenko et al. 2000