Análisis sobre prácticas de farmacología con animales de laboratorio: una mirada desde los estudiantes de la Universidad Nacional de San Luis, Argentina

Introduction: The subject Pharmacology in the Pharmacy program is theoretical and practical, lasts a semester, and is taught in the fourth year. Practical development involves six practical laboratory sessions (tpl) and four workshops. Three tpl are for in vivo animal experiments, two for computer programs and one for in vitro techniques, allowing students to evaluate the effect of drugs through experimentation, oriented towards biological and biomedical research, and to recognize pharmacological efficacy for the treatment of diseases. Our objective was to conduct a survey to obtain a guidance instrument that allows to increase teaching quality and to know perceptions and preferences about the practices concerned. Method: A cross-sectional, descriptive study was carried out and concluded in the first four-month period of the academic year 2015. Results: 90.48% reported that animal practices were useful for training, and two-thirds considered it necessary to implement alternatives to the use of animals in their education and training. 85.71% preferred in vivo experiments, 9.52% in vitro observations and 4.76% computer programs. 90.48% responded that they would work with laboratory animals again. Conclusions: We believe that it is appropriate to work with animals in this subject; it is important to consider this practical experience in pharmacology. Students expressed their interest in knowing and interpreting the effects and mechanisms of action of drugs through in vivo observations or organs manipulated by them.Introducción: la asignatura Farmacología en Farmacia es de carácter teórico-práctico, dura un cuatrimestre y se imparte en el cuarto año. El desarrollo práctico involucra seis trabajos prácticos de laboratorio (tpl) y cuatro talleres. En tres tpl se realiza experimentación in vivo con animales, en otros dos se trabaja con programas computacionales, y en el restante técnicas in vitro, permitiendo al estudiante valorar el efecto de los fármacos mediante la experimentación, orientado a la investigación biológica y biomédica, además de reconocer la eficacia farmacológica para el tratamiento de enfermedades. Nuestro objetivo fue promover una encuesta para obtener un instrumento orientativo que permita aumentar la calidad de la enseñanza y conocer las percepciones y preferencias sobre las prácticas en cuestión. Metodología: se realizó un estudio transversal y descriptivo, concluido el primer cuatrimestre académico del ciclo lectivo 2015. Resultados: el 90,48 % aludió que las prácticas con animales resultaron útiles para su formación, y las dos terceras partes consideró necesario implementar alternativas al uso de animales en aspectos relacionados a su educación y formación. El 85,71 % prefirió las observaciones in vivo; 9,52 % las in vitro y 4,76% los programas computacionales. 90,48 % respondió que volvería a trabajar con animales de laboratorio. Conclusiones: creemos que es pertinente trabajar con animales en esta asignatura, es importante pensar en esta experiencia práctica en la farmacología. Los estudiantes manifiestan su interés por conocer e interpretar los efectos y mecanismos de acción de los fármacos mediante observaciones in vivo o de órganos manipulados por ellos mismos

Structural Properties of Macrodontain I, a Cysteine Protease from Pseudananas macrodontes (Morr.) Harms (Bromeliaceae)

The primary structure of macrodontain I, a peptidase from Pseudananas macrodontes fruits, was determined using Edman's degradation. The enzyme is a non-glycosylated peptidase composed by 213 amino acids with a calculated molecular weight of 23,486.18 Da, pI value 6.99, and a molar extinction coefficient at 280 nm of 61,685 M-1 cm-1. The alignment of the sequence of macrodontain I with those cysteine peptidases from species belonging to the family Bromeliaceae showed the highest identity degree (87.74%) against fruit bromelain. A remarkable fact is that all these peptidase sequences show two Met contiguous residues (Met121 and 122) and the nonapeptide VPQSIDWRD located in the mature N-terminal region. Residues Cys26 and His159, which constitute the catalytic dyad in all cysteine peptidases, as well as active site residues Gln20 and Asn176, characteristic of Clan C1A, are conserved in macrodontain I. The 3-D model suggests that the enzyme belongs to the α + β class of proteins, with two disulfide bridges (Cys23-Cys63 and Cys57-Cys96) in the α domain, while the β domain is stabilized by another disulfide bridge (Cys153-Cys201). Further, we were able to establish that the cysteine peptidases from P. macrodontes are involved in the anti-inflammatory activity.Facultad de Ciencias Exacta

Effect of exogenous melatonin on acute and chronic inflammatory process in rats

Se investigó la influencia de melatonina (4 mg/kg) en la inflamación aguda usando el modelo de edema de pata en rata y en la inflamación crónica mediante la prueba del granuloma y artritis inducida por adyuvante. Melatonina inhibió el edema producido por carragenina en el modelo agudo, pero no presentó acción inhibitoria en la fase proliferativa de la prueba del granuloma y en la fase aguda y crónica de la artritis inducida por adyuvante. Estos resultados sugieren que melatonina, a la misma dosis, muestra diferentes comportamientos en los modelos inflamatorios ensayados.Melatonin influence (4 mg/kg) was investigated on acute inflammation using rat paw edema and on chronic inflammation through granuloma test and adjuvant arthritis. Melatonin inhibited the edema produced by carrageenan in acute model. However, failed to inhibit the proliferative phase in the granuloma test and the acute and chronic phase in the adjuvant arthritis. These results suggest melatonin shows different activity on the tested inflammatory models at the same doses.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Effect of exogenous melatonin on acute and chronic inflammatory process in rats

Se investigó la influencia de melatonina (4 mg/kg) en la inflamación aguda usando el modelo de edema de pata en rata y en la inflamación crónica mediante la prueba del granuloma y artritis inducida por adyuvante. Melatonina inhibió el edema producido por carragenina en el modelo agudo, pero no presentó acción inhibitoria en la fase proliferativa de la prueba del granuloma y en la fase aguda y crónica de la artritis inducida por adyuvante. Estos resultados sugieren que melatonina, a la misma dosis, muestra diferentes comportamientos en los modelos inflamatorios ensayados.Melatonin influence (4 mg/kg) was investigated on acute inflammation using rat paw edema and on chronic inflammation through granuloma test and adjuvant arthritis. Melatonin inhibited the edema produced by carrageenan in acute model. However, failed to inhibit the proliferative phase in the granuloma test and the acute and chronic phase in the adjuvant arthritis. These results suggest melatonin shows different activity on the tested inflammatory models at the same doses.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Anti-inflammatory Activity of Bromelia hieronymi: Comparison with Bromelain

Some plant proteases (e. g., papain, bromelain, ficin) have been used as anti-inflammatory agents for some years, and especially bromelain is still being used as alternative and/or complementary therapy to glucocorticoids, nonsteroidal antirheumatics, and immunomodulators. Bromelain is an extract rich in cysteine endopeptidases obtained from Ananas comosus. In this study the anti-inflammatory action of a partially purified extract of Bromelia hieronymi fruits, whose main components are cysteine endopeptidases, is presented. Different doses of a partially purified extract of B. hieronymi were assayed on carrageenan-induced and serotonine-induced rat paw edema, as well as in cotton pellet granuloma model. Doses with equal proteolytic activity of the partially purified extract and bromelain showed significantly similar anti-inflammatory responses. Treatment of the partially purified extract and bromelain with E-64 provoked loss of anti-inflammatory activity on carrageenan-induced paw edema, a fact which is consistent with the hypothesis that the proteolytic activity would be responsible for the anti-inflammatory action.Fil: Errasti, María Eugenia. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigacion de Proteinas Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Caffini, Nestor Oscar. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigacion de Proteinas Vegetales; ArgentinaFil: Pelzer, Lilian Eugenia. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; ArgentinaFil: Rotelli, Alejandra Ester. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; Argentin

Evaluation of Anti-Inflammatory Activity of Pseudananas macrodontes (Morr.) Harms (Bromeliaceae) Fruit Extract in Rats

Several species of the family Bromeliaceae are characterized by the production of proteases in unusual amounts, especially in fruits. Bromelain, an extract rich in cysteine endopeptidases obtained from Ananas comosus L., and a few other proteases have been used as anti-infl ammatory agents for some years, but bromelain is still mainly being used as alternative and/or complementary therapy to the treatment with glucocorticoids, nonsteroidal antirheumatics, and immunomodulators. In this study, the anti-infl ammatory action of a partially purifi ed extract from Pseudananas macrodontes (Morr.) Harms fruits (PPEPm) is presented, whose main components are cysteine endopeptidases. The effect of PPEPm was assessed in carrageenan-induced and serotonin-induced rat paw edema, as well as in the cotton pellet granuloma model. Doses with equal proteolytic activity of PPEPm and bromelain produced signifi cantly similar anti-infl ammatory responses in the acute infl ammatory models assayed, supporting the hypothesis that proteolytic activity could be responsible for the anti-infl ammatory action. On the contrary, comparable anti-infl ammatory effects of PPEPm and bromelain in the chronic infl ammatory assay required a much lower proteolytic activity content of PPEPm, which could be due to a differential affi nity for the protein target involved in this process.Fil: Errasti, María Eugenia. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigacion de Proteinas Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Caffini, Nestor Oscar. Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biologicas. Laboratorio de Investigacion de Proteinas Vegetales; ArgentinaFil: Pelzer, Lilian Eugenia. Universidad Nacional de San Luis; ArgentinaFil: Rotelli, Alejandra Ester. Universidad Nacional de San Luis; Argentin

Anti-inflammatory Activity and Antioxidant Potential of Aqueous Extracts from Stem Bark of Geoffroea decorticans

In the present work we investigated, for the first time, the anti-inflammatory activity and the antioxidant properties ofaqueous and ethanolic extracts, obtained from stem bark of Geoffroea decorticans (Gill. ex Hook. et Arn.) Burk. (Fabaceae). G.decorticans, commonly known as ?chañar? or ?chañarcillo?, is a traditional argentinean plant used as emollient, balsamic, antitussive,expectorant and anti-inflammatory. The stem bark was collected from San Francisco del Monte de Oro, San Luis, Argentina.Anti-inflammatory activity was evaluated by carrageenan-induced paw edema in rats. Antioxidant activity was tested using1,1-Diphenyl-2-picrylhydrazyl radical-scavenging activity (DPPH), 2,2´-Azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) radicalscavenging activity (ABTS) and ferric ion-reducing power (RP-Fe) assays. Aqueous extract 10% p/v showed anti-inflammatory activity (3h, 48% inhibition, 5h 37% inhibition and 7h 17% inhibition) and antioxidant activity (DPPH, IC50 (mg/mL) = 0.098 ± 0.032; ABTS, IC50 (mg/mL) = 0.022 ± 0.343, RP-Fe IC50 (mg/mL) = 1.124 ± 0.146). In the other hand, the ethanolic extract 5% p/v, presented anti-inflammatory activity (3h, 34% inhibition, 5h 38% inhibition and 7h 35% inhibition) and antioxidant activity (DPPH, IC50 (mg/mL) = 0.133 ± 0.027; ABTS, IC50 (mg/mL) = 0.086 ± 0.262, RP-Fe IC50 (mg/mL) = 7.089 ± 0.104). These results suggest that, also fruits, the aqueous and ethanolic extracts from the stem bark of G. decorticans present significant anti-inflammatory activity and antioxidant properties.Fil: Jofré, Mariana. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Area de Farmacognosia; ArgentinaFil: Fusco, María. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Area de Farmacognosia; ArgentinaFil: Favier, Laura. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Química. Area de Química Orgánica; ArgentinaFil: Teves, Mauricio. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Area de Farmacognosia; ArgentinaFil: Rotelli, Alejandra Ester. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Area de Farmacognosia; ArgentinaFil: Pedernera, Ana. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Area de Farmacognosia; ArgentinaFil: Pelzer, Lilian. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Area de Farmacognosia; ArgentinaFil: Ortega, Claudia. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Area de Farmacognosia; ArgentinaFil: Cifuente, Diego Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis; Argentina. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Departamento de Farmacia. Area de Farmacognosia; Argentin

Structural Properties of Macrodontain I, a Cysteine Protease from Pseudananas macrodontes (Morr.) Harms (Bromeliaceae)

The primary structure of macrodontain I, a peptidase from Pseudananas macrodontes fruits, was determined using Edman’s degradation. The enzyme is a non-glycosylated peptidase composed by 213 amino acids with a calculated molecular weight of 23,486.18 Da, pI value 6.99, and a molar extinction coefficient at 280 nm of 61,685 M−1 cm−1. The alignment of the sequence of macrodontain I with those cysteine peptidases from species belonging to the family Bromeliaceae showed the highest identity degree (87.74%) against fruit bromelain. A remarkable fact is that all these peptidase sequences show two Met contiguous residues (Met121 and 122) and the nonapeptide VPQSIDWRD located in the mature N-terminal region. Residues Cys26 and His159, which constitute the catalytic dyad in all cysteine peptidases, as well as active site residues Gln20 and Asn176, characteristic of Clan C1A, are conserved in macrodontain I. The 3-D model suggests that the enzyme belongs to the α + β class of proteins, with two disulfide bridges (Cys23-Cys63 and Cys57-Cys96) in the α domain, while the β domain is stabilized by another disulfide bridge (Cys153-Cys201). Further, we were able to establish that the cysteine peptidases from P. macrodontes are involved in the anti-inflammatory activity.Fil: Errasti, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Instituto Nacional de Tecnología Industrial. Centro de Investigación y Desarrollo del Cuero. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Centíficas. Centro de Investigación y Desarrollo del Cuero; ArgentinaFil: Natalucci, Claudia Luisa. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; ArgentinaFil: Caffini, Nestor Oscar. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Laboratorio de Investigación de Proteínas Vegetales; ArgentinaFil: Rotelli, Alejandra Ester. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia; ArgentinaFil: Brullo, Adriana. Università degli studi di Roma "La Sapienza"; ItaliaFil: Maras, Bruno. Università degli studi di Roma "La Sapienza"; ItaliaFil: Trejo, Sebastian Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Lopez, Laura Maria Isabel. Instituto Nacional de Tecnología Industrial. Centro de Investigación y Desarrollo del Cuero. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Centíficas. Centro de Investigación y Desarrollo del Cuero; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentin