La hipertensión arterial en animales de laboratorio / High blood pressure in experimental animals

Las Ciencias Médicas y Biológicas requieren, prioritariamente, que la investigación y la experimentación sean desarrolladas sobre organismos completos (los modelos animales). Su utilización ha permitido desarrollar innumerables ensayos preclínicos para evaluar los mecanismos patógenos y terapéuticos de diversas enfermedades, así como el estudio de las causas, naturaleza y cura de múltiples desórdenes de la salud humana. En este trabajo se muestra una panorámica general de los biomodelos de hipertensión arterial donde se describen: conceptos, características, origen, importancia, utilidad y procedimientos experimentales durante su fase de inducción. También se pondera la justificación de los biomodelos empleados en los estudios preclínicos de esta enfermedad. De igual forma, se describen los antecedentes para medir las alteraciones, las técnicas y los métodos directos e indirectos de medición de la presión arterial, la cual fue provocada experimentalmente en los animales de laboratorio para realizar los estudios de hipertensión humana.

Medical and biological sciences require, as a priority, that research and experimentation be carried out on complete organisms (animal models). Its use has allowed the development of innumerable preclinical tests to evaluate pathogenic and therapeutic mechanisms of various diseases, as well as to study the causes, nature and cure of multiple human health disorders. In this work, we show a general overview of arterial hypertension biomodels where concepts, characteristics, origin, importance, utility and experimental procedures during their induction phase are described. The justification of the biomodels used in preclinical studies of this disease is also considered. Antecedents are also described to measure alterations, techniques and direct and indirect methods of measurement of arterial pressure, which was provoked experimentally in the laboratory animals to carry out the studies of human hypertension.

Effect of Cissus quadrangularis on Fracture Healing in Laboratory Animal / Efeito do Cissus quadrangularis na cicatrização de fraturas em animais de laboratório

Objectives: The present study was designed to examine the efficacy of Cissus quadrangularis paste on fracture healing in artificially induced fractured rabbits. Methods: Fifteen rabbits were separated into three groups namely A, B, and C. Veldt grape paste was applied in groups B and C (Treatment Group) by close reduction and open reduction methods of fracture management; respectively, while group A was kept as control. The blood parameter and fracture healing properties in all animals have been monitored and examined routinely during the study period. Results: Both treated groups revealed lower serum calcium levels (SCL) than the control group after 24 hours of fracture that became within the normal range on the 14th day. Fracture healing in the treated groups has been commenced more rapidly than the control group with complete bridging of discontinuity by a distinct osseous callus in the fracture line on day 7 and complete effacing of fracture line on day 14. Conclusion: We did not find any type of anomalousness, clinical deviations, and alteration of serum calcium level on the 14th day of the fracture in treated animals hence Veldt Grape paste could be readily applicable to the management of the fracture in animals.

Objetivos: avaliar a eficácia da pasta de Cissus quadrangularis na consolidação de fraturas em coelhos fraturados artificialmente. Metodos: quinze coelhos foram separados em três grupos (A, B e C). A pasta foi aplicada nos grupos B e C (Grupo de Tratamento) pelos métodos de redução fechada e redução aberta de gerenciamento de fraturas; respectivamente. O grupo A foi mantido como controle. O parâmetro sanguíneo e as propriedades de cicatrização de fraturas em todos os animais foram monitorados e examinados, rotineiramente, durante o período do estudo. Resultados: ambos os grupos tratados revelaram níveis séricos de cálcio (SCL) mais baixos do que o grupo controle, após 24 horas de fratura que se tornou normal no 14º dia. A cicatrização da fratura nos grupos tratados foi iniciada mais rapidamente do que o grupo controle, com ponte completa de descontinuidade por um calo ósseo distinto na linha de fratura no dia 7 e apagamento completo da linha de fratura no dia 14. Conclusao: não encontramos nenhum tipo de anomalia, desvios clínicos e alteração do nível sérico de cálcio no 14º dia da fratura nos animais tratados, portanto, a pasta Veldt Grape pode ser aplicável ao manejo da fratura em animais.

Efectos de la inyección neonatal de estreptozotocina sobre indicadores bioquímicos y de estrés oxidativo en ratas / Effects of neonatal injection of streptozotocin on biochemical indicators and oxidative stress in rats

Introducción: Debido a sus propiedades químicas, la estreptozotocina es uno de los agentes diabetogénicos más utilizados para generar modelos biológicos de diabetes, por lo que es necesario estudiar cuáles son sus efectos en el organismo del animal de laboratorio. Objetivo: Evaluar, en un periodo de 90 días, los efectos de la inyección neonatal de estreptozotocina en ratas Wistar sobre indicadores bioquímicos y de estrés oxidativo en hígado y riñón. Métodos: La diabetes fue inducida neonatalmente por 100 mg de estreptozotocina en ratas Wistar. Se realizaron determinaciones de glucemia, insulina e indicadores de estrés oxidativos en hígado y riñón en cinco animales por grupo a los días 5, 10, 20, 30, 60, 90 de nacidos. Resultados: En todas las intervenciones, la glucemia e insulina mostraron diferencias significativas en el grupo-STZ respecto al control. El valor máximo de hiperglucemia se observó al quinto día. La concentración de nitratos y nitritos en hígado fue mayor que en riñón. En comparación con el grupo control, en el tejido hepático del grupo-STZ la concentración de nitratos y nitritos resultó significativamente superior los días 10-20. En todas las intervenciones se detectó consumo de glutatión reducido en ambos órganos. En el hígado de las ratas STZ no se demostró daño a lípidos ni proteínas; sin embargo, en riñón se detectó daño significativo en ambas biomoléculas al quinto día. Conclusiones: Tanto la citotoxicidad de la estreptozotocina neonatal como las concentraciones de glucosa e insulina inducidas repercutieron negativamente sobre los indicadores de estrés oxidativo estudiados en tejido hepático y renal(AU)

Introduction: Streptozotocin is currently one of the most used diabetogenic agents to generate biological models of diabetes due to its chemical properties, so it is necessary to study the consequences of STZ for the organism of the laboratory animal. Objective: To evaluate in a period of 90 days the effects of neonatal injection of streptozotocin in Wistar rats on biochemical indicators and oxidative stress in liver and kidney. Methods: Diabetes was induced neonatally by 100 mg of streptozotocin in Wistar rats. Blood glucose, insulin and oxidative stress indicators in liver and kidney were determined in 5 animals per group at days 5, 10, 20, 30, 60, 90 of birth. Results: Blood glucose and insulin showed significant differences in the STZ-group respect to the control group in all interventions. The maximum value of hyperglycemia was observed on day-5. The concentration of nitrates and nitrites in liver was higher than in kidney. In liver tissue of the STZ-group, this indicator was significantly higher on days 10-20 compared to the control. In all interventions, reduced glutathione consumption was demonstrated in the STZ-group compared to control in both organs. In the liver of STZ rats no lipid or protein damage was demonstrated. However, in the kidney, significant damage in both biomolecules was detected in the STZ-group on day-5. Conclusions: Neonatal streptozotocin cytotoxicity as well as induced glucose and insulin concentrations had a negative impact on oxidative stress indicators studied in liver and kidney tissue(AU)

Ethical considerations in animal research: the principle of 3r's

In the last century, progress in the knowledge of human diseases, their diagnosis and treatment have grown exponentially, due in large part to the introduction and use of laboratory animals. Along with this important progress, the need to provide training and guidance to the scientific community in all aspects related to the proper use of experimental animals has been indispensable. Animal research committees play a primary role in evaluating experimental research protocols, from their feasibility to the rational use of animals, but above all in seeking animal welfare. The Institutional Committee for the Care and Use of Animals (IACUC) has endeavored to share several relevant aspects in conducting research with laboratory animals. Here, we present and discuss the topics that we consider of utmost importance to take in the account during the design of any experimental research protocol, so we invite researchers, technicians, and undergraduate and graduate students to dive into the fascinating subject of proper animal care and use for experimentation. The main intention of these contributions is to sensitize users of laboratory animals for the proper and rational use of them in experimental research, as well as to disseminate the permitted and unpermitted procedures in laboratory animals. In the first part, the significance of experimental research, the main functions of IACUC, and the principle of the three R's (replacement, reduction, and refinement) are addressed.

Allogenic platelet-rich plasma in induced ulcers in rat's cornea / [Plasma rico em plaquetas alogênico em úlceras de córnea induzidas em ratos]

The objective in this study was to evaluate the clinic effect of applying allogenic platelet-rich plasma (PRP) heated or not, for treating cornea ulcers, including the dosage of PDGF-BB in the cornea. The ulcers were induced, standardizing the left eye from 81 rats (Ratus norvegicus, albinus variety), assigned randomly into three groups (N=27): control group (CG) which did not receive any topic treatment; heated PRP group (GA) and PRP group (GP), which received topical treatment every eight hours for five days. Each group underwent evaluation at 24 hours (M1), three days (M3) and five days (M5). The clinical exam evaluated the opacity, vascularization and corneal repair. The corneal PDGF-BB was dosed through the ELISA method. The corneal opacity was decreased in PRP-treated animals (GA and GP) and corneal repair time reduced when compared to CG at M1 and M5. Furthermore, GP showed greater vascularization at M3 compared to M1. Applied allogenic PRP eye drops, heated or not, speed up corneal healing, and reduce corneal repair time. However, the corneal PDGF concentration was not altered in any of the treatments.(AU)

Objetivou-se avaliar o efeito clínico da aplicação de plasma rico em plaquetas alogênico (PRP) aquecido ou não, no tratamento de úlceras de córnea, como a dosagem de PDGF-BB na córnea. As úlceras foram induzidas, padronizando-se o olho esquerdo de 81 ratos (Rattus norvegicus, variedade albinus), aleatoriamente, nos três grupos (N = 27): grupo controle (CG), que não recebeu nenhum tratamento tópico; grupo PRP aquecido (GA) e grupo PRP (GP), que receberam tratamento tópico a cada oito horas, durante cinco dias. Cada grupo foi subdividido em 24 horas (M1), três dias (M3) e cinco dias (M5). O exame clínico avaliou a opacidade, a vascularização e o reparo corneano. O PDGF-BB corneano foi dosado pelo método Elisa. Houve diminuição da opacidade da córnea nos animais tratados com PRP (GA e GP) e diminuição do tempo de reparo da córnea em comparação com CG, M1 e M5. Além disso, foi observada maior vascularização no GP no momento M3 em relação ao M1. A aplicação de colírios de PRP alogênico, aquecidos ou não, acelera a cicatrização da córnea, além de reduzir o tempo de reparo da córnea. No entanto, a concentração de PDGF na córnea não se alterou em nenhum dos tratamentos.(AU)

Protocolo de prueba de ventilador mecánico para atención de pacientes con COVID-19 en modelo porcino / Mechanical ventilator test protocol for care of patients with COVID-19 in porcine model

RESUMEN A raíz de la pandemia generada por el SARS-CoV-2, los sistemas de salud de los distintos países han experimentado un dramático aumento en la ocupación de las unidades de cuidado intensivo y, por ende, una alta demanda de ventiladores mecánicos. En este artículo se propone un protocolo de pruebas de ventiladores mecánicos de bajo costo en modelos porcinos, como parte del desarrollo de equipos biomédicos que pueden apoyar la crisis sanitaria suscitada por la pandemia. El protocolo aborda aspectos que incluyen la fase previa a la prueba, el manejo anestésico, de vía aérea, seguimiento de laboratorio, recuperación y seguimiento del animal. Como resultado, se presenta a la comunidad académica un protocolo de validación de prototipo de ventilador mecánico en espécimen porcino como un referente para su aplicación y revalidación por parte de grupos interesados en el desarrollo de tecnologías locales de bajo costo.

ABSTRACT As a result of the SARS-CoV-2 pandemic, health systems globally have seen a dramatic increase in the occupancy of intensive care units, with mechanical ventilators being a resource in high demand in the care of these patients. This article proposes a protocol for testing low-cost mechanical ventilators in pig models, as part of the development of locally manufactured biomedical equipment that can support the health crisis caused by the pandemic. The protocol addresses aspects that include the pre-test phase, anesthetic and airway management, laboratory monitoring, recovery and monitoring of the animal. This document presents to the academic community a validation protocol of a mechanical ventilator prototype in a pig specimen that can be a reference for its application and revalidation by other groups interested in the development of local and low-cost technologies.

Protocolo de prueba de ventilador mecánico para atención de pacientes con COVID-19 en modelo porcino / Mechanical ventilator test protocol for care of patients with COVID-19 in porcine model

RESUMEN A raíz de la pandemia generada por el SARS-CoV-2, los sistemas de salud de los distintos países han experimentado un dramático aumento en la ocupación de las unidades de cuidado intensivo y, por ende, una alta demanda de ventiladores mecánicos. En este artículo se propone un protocolo de pruebas de ventiladores mecánicos de bajo costo en modelos porcinos, como parte del desarrollo de equipos biomédicos que pueden apoyar la crisis sanitaria suscitada por la pandemia. El protocolo aborda aspectos que incluyen la fase previa a la prueba, el manejo anestésico, de vía aérea, seguimiento de laboratorio, recuperación y seguimiento del animal. Como resultado, se presenta a la comunidad académica un protocolo de validación de prototipo de ventilador mecánico en espécimen porcino como un referente para su aplicación y revalidación por parte de grupos interesados en el desarrollo de tecnologías locales de bajo costo.

ABSTRACT As a result of the SARS-CoV-2 pandemic, health systems globally have seen a dramatic increase in the occupancy of intensive care units, with mechanical ventilators being a resource in high demand in the care of these patients. This article proposes a protocol for testing low-cost mechanical ventilators in pig models, as part of the development of locally manufactured biomedical equipment that can support the health crisis caused by the pandemic. The protocol addresses aspects that include the pre-test phase, anesthetic and airway management, laboratory monitoring, recovery and monitoring of the animal. This document presents to the academic community a validation protocol of a mechanical ventilator prototype in a pig specimen that can be a reference for its application and revalidation by other groups interested in the development of local and low-cost technologies.

Oogenic development and gonotrophic cycle of Aedes aegypti and Aedes albopictus in laboratory / Desarrollo oogénico y ciclo gonotrófico de Aedes aegypti y Aedes albopictus en laboratorio

Abstract: Objective: To determine the time of oogenic development and the length of the gonotrophic cycle of Ae. aegypti and Ae. albopictus in laboratory. Materials and methods: Bloodfed females of Ae. aegypti and Ae. albopictus were dissected every 4 h to determine the development status of the follicles according to the Christophers' stages. Results: The minimum time of oocyte maturation in Ae. aegypti and Ae. albopictus was 64-82 h and 52-64 h post-feeding, respectively. We found that the gonotrophic cycle of Ae. aegypti (3.7-4.2 d) is longer than that of Ae. albopictus (3.2-3.7 d). The follicle length showed significant differences between species at Christophers' stages 2" and 5, whereas follicle amplitude was different between the two mosquitoes at stages 2", 3 and 4. Conclusions: The study provided new evidence on the reproductive strategies of Ae. aegypti and Ae. albopictus females that coexist in the Neotropical region of Mexico.

Resumen: Objetivo: Determinar el tiempo de desarrollo oogénico y del ciclo gonotrófico de Aedes aegypti y Aedes albopictus en laboratorio. Material y métodos: Hembras de Ae. aegypti y Ae. albopictus alimentadas con sangre fueron disecadas cada cuatro horas para determinar el estado de desarrollo folicular, según los estadios de Christophers. Resultados: El tiempo mínimo de maduración del oocito en Ae. aegypti y Ae. albopictus fue de 64-82 h y 52-64 h post-alimentación, respectivamente. El ciclo gonotrófico de Ae. aegypti (3.7-4.2 d) fue mayor que el de Ae. albopictus (3.2-3.7 d). La longitud folicular presentó diferencias significativas entre las especies en los estadios de Christophers 2" y 5, mientras que la amplitud folicular fue diferente entre ambos mosquitos en los estadios 2", 3 y 4. Conclusiones: El estudio proporcionó nueva evidencia sobre la estrategia reproductiva de las hembras de Ae. aegypti y Ae. albopictus que coexisten en la región neotropical de México.

Laboratorio de Cirugía Experimental: innovación tecnológica al servicio de la asistencia, la docencia e investigación / Laboratory of Experimental Surgery: technological innovation at the service of teaching, research and medical assistance

RESUMEN El Laboratorio de Cirugía Experimental de la Universidad de Ciencia Médicas de Villa Clara cuenta con la tecnología necesaria para el montaje y desarrollo de técnicas y habilidades quirúrgicas y microquirúrgicas, las cuales son utilizadas por los profesionales y estudiantes en la docencia y la investigación biomédica. Se realizaron varias innovaciones con la recuperación de equipos, instrumentos y materiales en desuso; se creó el gimnasio quirúrgico conformado por modelos inanimados sintéticos, y se implementó el modelo «pollo¼, como técnica alternativa al uso de animales vivos en la experimentación animal. Se han desarrollado: importantes proyectos de investigación, entrenamientos de cirugía y microcirugía experimental, diversos programas docentes, y exitosos cursos internacionales con estudiantes y residentes. El laboratorio constituye un valioso aporte de alternativas éticas, económicas y sanitarias, que han favorecido la formación de recursos humanos y un servicio científico-técnico de calidad para la docencia y la investigación.

ABSTRACT The Laboratory of Experimental Surgery at the University of Medical Sciences of Villa Clara has the necessary technology for the assembly and development of skills, as well as, surgical and microsurgical techniques, which are used by professionals and students in teaching and biomedical research. Several innovations were made with the recovery of equipments, instruments and disused materials; the surgical gymnasium was made up of synthetic inanimate models, and the "chicken" model was implemented as an alternative technique to the use of live animals in animal experimentation. Important research projects, training in surgery and experimental microsurgery, diverse teaching programs, and successful international courses with students and residents have been developed. The laboratory constitutes a valuable contribution of ethical, economic and sanitary alternatives, which have favored the formation of human resources and a qualified scientific-technical service for teaching and research.

Evaluación genotóxica de las moléculas con efectos tripanocida y leishmanicida cantin-6-ona y 5-metoxicantin-6-ona en células de médula ósea de ratones / Genotoxic evaluation of the molecules with trypanocidal and leishmanicidal effects, canthin-6-one and 5-methoxycanthin-6-one in mouse bone marrow cells

La tripanosomiasis americana y la leishmaniasis son problemas de salud pública relevantes en Iberoamérica. Las drogas utilizadas actualmente para el tratamiento de estas enfermedades poseen efectos colaterales tóxicos severos. Varios grupos de investigación están abocados a la búsqueda de productos naturales y sintéticos para encontrar nuevos agentes terapéuticos efectivos que no presenten reacciones colaterales adversas. En la evaluación de compuestos de la especie vegetal Zanthoxylum chiloperone (Rutaceae), se demostró que compuestos aislados del extracto presentaban actividad leishmanicida, tripanocida y antifúngica in vivo. Teniendo como antecedentes estos resultados, en el presente estudio se evaluaron los efectos genotóxico y citotóxico del cantín-6-ona y del 5-metoxicantin-6-ona, moléculas aisladas de la planta, en células de médula ósea de animales tratados. El estudio de los efectos genotóxicos se hizo a través del ensayo de modificaciones en la frecuencia de micronúcleos y el efecto citotóxico por modificaciones en la relación entre eritrocitos policromáticos y eritrocitos normocromáticos. Se realizaron 2 ensayos independientes y en cada ensayo los animales fueron divididos en tres grupos de tratamiento: GRUPO I: control negativo que recibió 200 uL de agua y 2.1% de DMSO, vía oral, GRUPO II: compuesto a ser evaluado (canthin-6-ona o 5-methoxicantin-6-ona) con 2.1% de DMSO, y GRUPO III: control positivo que recibió ciclofosfamida 50mg/kg/peso del animal, vía intraperitoneal. El análisis estadístico mostró que ambos compuestos no presentaron efectos genotóxicos ni citotóxicos. Estos resultados permiten proponer a estas moléculas como candidatas a ser sometidas a estudios más detallados como potenciales fármacos contra estas dos enfermedades

American trypanosomiasis and leishmaniasis are relevant public health problems in Latin America. The drugs currently used to treat these diseases have severe toxic side effects. Several research groups are dedicated to the search of natural and synthetic products to find new effective therapeutic agents that do not present adverse collateral reactions. In the evaluation of compounds of the plant species Zanthoxylum chiloperone (Rutaceae), it was shown that isolated compounds of the extract had leishmanicidal, trypanocidal and antifungal in vivo activities. Based on these results, the genotoxic and cytotoxic effects of canthin-6-one and 5-methoxycanthin-6-one, molecules isolated from the plant, on bone marrow cells of treated mice were evaluated in the present study. The study of genotoxic effects was made through the test of modifications in the frequency of micronuclei and the cytotoxic effects by modifications in the relationship between polychromatic erythrocytes and normochromic erythrocytes. Two independent assays were performed and in each assay the animals were divided into three treatment groups: GROUP I: negative control that received 200 µL of water and 2.1% of DMSO, orally, GROUP II: compound to be evaluated (canthin-6 -one or 5-methoxycanthin-6-one) with 2.1% DMSO, and GROUP III: positive control that received cyclophosphamide 50mg /kg animal weight, intraperitoneal. Statistical analysis showed that both compounds had neither genotoxic nor cytotoxic effects. These results allow these molecules to be proposed as candidates to be subjected to more detailed studies as potential drugs against these two diseases

Leyendo a Spallanzani hoy en día / Reading Spallanzani today

Resumen Recordamos al abate Lázaro Spallanzani (1729-1799) fundamentalmente por su victoriosa contienda con Needham sobre la generación espontánea, pero fue un hombre de múltiples intereses en distintos campos de la ciencia, desde la biología a la volcanología. Se lo llamó el "biólogo de los biólogos", desarrollando una serie de investigaciones sobre la reproducción de los anfibios, en una de las cuales - "Experiencias al servicio de la historia de la generación de animales y plantas" - el lector moderno se estremece de horror ante las crueles mutilaciones que infligiera a los sapos machos tratando de interrumpir su copulación con las hembras. Él mismo las califica de "bárbaras" estas torturas, inadmisibles en un hombre que detentaba una jerarquía eclesiástica, aún en una época como la suya, en que se calificaba a los animales como "anima vili" (cosa sin valor). Mucho se ha avanzado hoy en día en resguardo de nuestros "hermanos menores", como los llamara San Francisco, pero aún falta mucho que hacer en la regulación de la experimentación científica que los utiliza.

Abstract We remember Lazaro Spallanzani (1729-1799) mainly for his controversy with Needham over spontaneous generation, but he was a man of multiple scientific activities in the fields of biology, mineralogy, physics, mathematics and… volcanology! Called "the biologist of biologists", he developed a series of investigations about reproduction of amphibian, in one of them -Experiences in service to the history of the generation of animals and plants- we have found horrific experiments with frogs, including severe and useless mutilation of males, in order to interrupt its copulation with females, acts he describes as "barbaric", and we estimate inadmissible in the ecclesiastic man he was, even in an epoch in which animals were considered "anima vili" (something without value). A brief review of the use of animals in laboratories shows significant advances in the ethical regulations for this practice, but we believe that these achievements are not enough.

Evaluation by fractal dimension of muscle regeneration after photobiomodulation / Avaliação por meio da dimensão fractal da regeneração muscular após fotobiomodulação

Abstract Introduction: Many treatment modalities are used for muscle tissue recovery. Photobiomodulation is a modality that can be employed to improve the quality of tissue repair. The use of fractal dimension (FD) is an innovative methodology in the quantitative evaluation of treatment efficacy. Objective: Use FD as a quantitative analysis method to evaluate the effect of photobiomodulation of 904 nanometers (nm) in the initial phase of the muscle regeneration process. Method: Thirty male Wistar rats were divided into three groups: Control Group (CG), Injured and Untreated Group (IUT), and Injured and Treated Group (IT). Muscle injury was induced by cryoinjury in the central region of the anterior tibial (AT) belly of the left posterior limb. This was performed by an iron rod that was previously immersed in liquid nitrogen. Applications started 24 hours after the injury and occurred daily for five days. They were performed at two points in the lesion area. The rats were euthanized on the seventh day. The AT muscles were removed and frozen in liquid nitrogen. Then, the histological sections were stained using the Hematoxylin-Eosin (HE) technique and submitted to FD analysis performed by the box-counting method using ImageJ software. The Kolmogorov-Smirnov test was used for data normality, and the Kruskall-Wallis test and Dunn's post-test were used for group comparison (p<0.05%). Results: Differences between IT and IUT groups were statistically significant, and it was possible to observe the reduction of fractability with p=0.0034. Conclusion: FD is a useful tool for the analysis of skeletal muscle disorganization in the initial phase of regeneration and confirms the potentially beneficial effects of photobiomodulation to this process.

Resumo Introdução: Diversas modalidades de tratamento são utilizadas para recuperação do tecido muscular, dentre elas a fotobiomodulação pode ser empregada para melhorar a qualidade da regeneração e a dimensão fractal se apresenta como uma metodologia inovadora na avaliação quantitativa da eficácia do tratamento. Objetivo: Utilizar a dimensão fractal como método de análise quantitativa do efeito do Laser de Arseneto de Gálio (AsGa) na fase inicial do processo de regeneração muscular. Método: Foram utilizados trinta ratos Wistar, machos divididos em: Grupo Controle (CT), Grupo lesado e não tratado (LNT) e Grupo Lesado e tratado (LT). A lesão muscular foi induzida por criolesão na região central do ventre do músculo tibial anterior (TA) do membro posterior esquerdo, por meio de uma haste de ferro previamente imersa em nitrogênio líquido. As aplicações foram iniciadas 24 horas após a lesão, diariamente, durante cinco dias, em dois pontos na área da lesão. No sétimo dia os animais foram eutanasiados; o músculo TA retirado, congelado em nitrogênio líquido e os cortes histológicos corados com a técnica de Hematoxilina-Eosina para serem então submetidos à análise de dimensão fractal realizada pelo método boxcounting através do software Image J. Para a normalidade dos dados utilizou-se Kolmogorov Smirnov, para as comparações teste de Kruskall-Wallis com pós teste de Dunn (p<0,05%). Resultados: A comparação entre LT e LNT foi estatisticamente significativa, sendo possível observar a redução da fractabilidade com p=0,0034. Conclusão: A dimensão fractal é uma ferramenta útil para análise da desorganização músculo esquelética na fase inicial da regeneração e mostra o potencial efeito benéfico da fotobiomodulação nesse processo.

Conhecimento sobre a bioética e a Lei 11. 794/2008 na graduação / The knowledge of bioethics and Law 11, 794/2008 in undergraduate courses / Conocimiento sobre bioética y la Ley 11. 794/2008 en el grado

Resumo O uso de animais para fins didáticos e de pesquisa requer cuidados específicos. Atualmente, vigora no Brasil a Lei 11.794/2008, que rege parâmetros legais de manejo e conduta neste caso. Esta lei foi acompanhada da instalação ou adequação de comissões de ética em instituições que utilizam animais para ensino e investigação, bem como da criação do Conselho Nacional de Controle de Experimentação Animal. No entanto, apesar dos avanços, especialmente na legislação, ainda não foi consolidada nenhuma grande mudança de comportamento de pesquisadores e alunos de graduação que manuseiam animais em laboratório. A divulgação de informações deixa a desejar, e a prática acaba por repercutir a carência de reflexão ética. Este artigo busca averiguar o atual conhecimento bioético de alunos de graduação e professores com o objetivo de estimular mudanças de conduta. Aprovado CEP-Unioeste CAAE 8563417.8.0000.0107

Abstract Used for education and research, laboratory animals require special care on their handling. Brazilian Law 11,794/2008 establishes the legal parameters for animal manipulation and welfare. It was accompanied by the obligatory installation of the Institutional Ethics Committees on the Use of Animals and the creation of the National Council for Animal Experimentation Control. There have been advances in the field of animal bioethics legislation. However, considering the behavior of those who handle the animals in laboratory environment, especially undergraduate students, these advances are insufficient: the information does not reach them and their attitudes remain in need of ethical reflection. This article seeks to investigate the current bioethical knowledge of undergraduate students and teachers in order to stimulate changes in conduct. Aprovado CEP-Unioeste CAAE: 78563417.8.0000.0107

Resumen El uso de animales con fines didácticos y de investigación requiere cuidados específicos. Actualmente, rige en Brasil la Ley 11.794/2008 que regula los parámetros legales de manejo y conducta en estos casos. Esta ley estuvo acompañada de la instalación o adecuación de comisiones de ética en instituciones que utilizan animales para enseñanza e investigación, así como de la creación del Consejo Nacional de Control de Experimentación Animal. No obstante, a pesar de los avances, especialmente en la legislación, aún no se ha consolidado ninguna gran transformación en el comportamiento de los investigadores y alumnos que manipulan animales en el laboratorio. La divulgación de informaciones es insuficiente, y la práctica acaba reflejando la falta de reflexión ética. Este artículo procura identificar el conocimiento bioético actual de los alumnos de grado y de los profesores, con el objetivo de estimular cambios en la conducta. Aprovado CEP-Unioeste CAAE: 78563417.8.0000.0107

Machine-Learning Based Automatic and Real-time Detection of Mouse Scratching Behaviors

Scratching is a main behavioral response accompanied by acute and chronic itch conditions, and has been quantified as an objective correlate to assess itch in studies using laboratory animals. Scratching has been counted mostly by human annotators, which is a time-consuming and laborious process. It has been attempted to develop automated scoring methods using various strategies, but they often require specialized equipment, costly software, or implantation of device which may disturb animal behaviors. To complement limitations of those methods, we have adapted machine learning-based strategy to develop a novel automated and real-time method detecting mouse scratching from experimental movies captured using monochrome cameras such as a webcam. Scratching is identified by characteristic changes in pixels, body position, and body size by frame as well as the size of body. To build a training model, a novel two-step J48 decision tree-inducing algorithm along with a C4.5 post-pruning algorithm was applied to three 30-min video recordings in which a mouse exhibits scratching following an intradermal injection of a pruritogen, and the resultant frames were then used for the next round of training. The trained method exhibited, on average, a sensitivity and specificity of 95.19% and 92.96%, respectively, in a performance test with five new recordings. This result suggests that it can be used as a non-invasive, automated and objective tool to measure mouse scratching from video recordings captured in general experimental settings, permitting rapid and accurate analysis of scratching for preclinical studies and high throughput drug screening.

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No abstract available.

Optical coherence tomography angiography in preclinical neuroimaging

Preclinical neuroimaging allows for the assessment of brain anatomy, connectivity, and function in laboratory animals, such as mice and this imaging field has been a rapidly growing aimed at bridging the translation gap between animal and human research. The progress in the animal research could be accelerated by high-resolution in vivo optical imaging technologies. Optical coherence tomography-based angiography (OCTA) estimates the scattering from moving red blood cells, providing the visualization of functional micro-vessel networks within tissue beds in vivo without a need for exogenous contrast agents. Recent advancement of OCTA methods have expanded its application to neuroimaging of small animal models of brain disorders. In this paper, we overview the recent development of OCTA techniques for blood flow imaging and its preclinical applications in neuroimaging. In specific, a summary of preclinical OCTA studies for traumatic brain injury, cerebral stroke, and aging brain on mice is reviewed.

comparative histological study of submucosal gland hypertrophy in trachea of mice exposed to cigarette and shisha smoke

Objective: To observe the effects of Shisha smoke on submucosal glands of trachea of mice; and compare it with tracheal glands of mice exposed to cigarette smoke

Study Design: Randomised controlled trial

Place and Duration of Study: Department of Anatomy, Islamic International Medical College, Rawalpindi, in collaboration with National Institute of Health [NIH], Islamabad from October 2013 till April 2014

Methodology: Sample comprised of 40 adult male mice of strain BALB/c. They were randomly divided into three groups. Control group was labelled as Group 'C'. The mice in this group were kept in a whole body smoke exposure chamber and were exposed to fresh air. Shisha group was labelled as Group 'SS', and the mice in this group were exposed to Shisha smoke. Mice in the third group labelled as Group CS were exposed to cigarette smoke. All the mice were dissected after an exposure period of eight weeks. Tracheal tissue was stained and examined microscopically for submucosal gland hypertrophy and compared with the control group, using Reid's Index. An Index of more than 0.4 is termed as hypertrophy

Results: There was significant submucosal gland hypertrophy in groups CS and SS as compared to group C. There was also significant difference in the frequency of mucosal hypertrophy between SS [93.7%] and CS groups [53.3%], which was found statistically significant [p<0.001]

Conclusion: Shisha smoking was significantly associated with mucosal hypertrophy when compared with cigarette smoking and controls. Shisha smoke contains higher level of toxicants as compared to cigarette smoke, and it causes more oxidative damage of tissues

Effects of bone marrow mesenchymal stem cells-conditioned medium on tibial partial osteotomy model of fracture healing in hypothyroidism rats

Background: Hypothyroidism is associated with dysfunction of the bone turnover with reduced osteoblastic bone formation and osteoclastic bone resorption. Mesenchymal stem cells [MSCs] secrete various factors and cytokines that may stimulate bone regeneration. The aim of this study was to determine the effects of MSCs-conditioned medium [CM] in hypothyroidism male rats after inducing bone defect

Methods: In this study, 24 male rats were randomly assigned to three groups: [I] hypothyroidism + bone defect [HYPO], [II] hypothyroidism + bone defect + CM [HYPO + CM], and [III] no hypothyroidism + bone defect [control]. Four weeks after surgery, the right tibia was removed, and immediately, biomechanical and histological examinations were performed

Results: The results showed a significant reduction in bending stiffness [32.64 +/- 3.99], maximum force [14.63 +/- 1.89], high stress load [7.59 +/- 2.31], and energy absorption [12.68 +/- 2.12] at the osteotomy site in hypothyroidism rats in comparison to the control and hypothyroidism + condition medium groups [p < 0.05]. There was also a significant decrease in the trabecular bone volume [3.86 +/- 3.88] and the number of osteocytes [5800 +/- 859.8] at the osteotomy site in hypothyroidism rats compared to the control and hypothyroidism + condition medium groups [p < 0.01 and p < 0.02, respectively]

Conclusion: The present study suggests that the use of the CM can improve the fracture regeneration and accelerates bone healing at the osteotomy site in hypothyroidism rats

Lipid peroxidation and its role in the expression of NLRP1a and NLRP3 genes in testicular tissue of male rats: a model of spinal cord injury

Background: The majority of male patients with spinal cord injury [SCI] suffer from infertility. Nucleotide-binding oligomerization domain-like receptors NOD-like receptors [NLRs] are a kind of receptors that corporate in the inflammasome complex. Recent studies have introduced the inflammasome as the responsible agent for secreting cytokines in semen. Reactive oxygen species [ROS] is one of the elements that trigger inflammasome activation. Genital infections in SCI can lead to ROS generation. We investigated the relation between lipid peroxidation and inflammasome complex activity in testicular tissue of SCI rats

Methods: Adult male rats [n=20], weighting 200- 250 g, were included and divided into four groups: three experimental groups, including SCI1, SCI3, and SCI7, i.e. the rats were subjected to SCI procedure and sacrificed after one, three, and seven days, respectively and a control group. We performed a moderate, midline spinal contusion injury at thoracic level 10. The animals were anesthetized, and testes were collected for measurement of gene expression by real-time PCR. Caudal parts of epididymis were collected for malondialdehyde [MDA] measurement

Results: No NLRP1a mRNA overexpression was seen in the testes of control and SCI groups. After seven days from SCI surgery, NLRP3 mRNA expression was significantly increased in SCI7 animals [p

Conclusion: NLRP3 overexpression occurs due to the increased ROS production in testis tissue of SCI rats

Nanocurcumin-mediated down-regulation of telomerase via stimulating TGF beta 1 signaling pathway in hepatocellular carcinoma cells

Background: Curcumin, extracted from turmeric, represents enormous potential to serve as an anticancer agent. Telomerase is viewed as a prominent molecular target of curcumin, and transforming growth factor- beta 1 [TGF beta 1] has proven to be a major inhibitory signaling pathway for telomerase activity. In the current study, we aimed to explore suppressive effects of nanocurcumin on telomerase expression through TGF beta 1 pathway in a hepatocellular carcinoma cell line [Huh7]

Methods: MTT assay was used to determine the effect of nonocurcumin on viability of Huh7 cells. RT-PCR was used to analyze the gene expression patterns

Results: MTT assay revealed that nanocurcumin acts in a dose- and time-dependent manner to diminish the cell viability. RT-PCR analysis indicated that nanocurcumin results in augmentation of TGF beta 1 72 hours post treatment and leads to the reduction of telomerase expression 48 and 72 hours post exposure. Also, up-regulation of Smad3 and E2F1 and down-regulation of Smad7 confirmed the effect of nanocurcumin on intermediate components of TGF beta 1 pathway. Furthermore, transfection of the proximal promoter of telomerase triggered a significant reduction in luciferase activity

Conclusion: The data from the present study lead us to develop a deeper understanding of the mechanisms underlying nanocurcumin-mediated regulation of telomerase expression, thereby presenting a new perspective to the landscape of using nanocurcumin as a cancer-oriented therapeutic agent