Conservation of Organs (Heart, Brain and Kidney) of Canine by Cold-Temperature Silicone Plastination in an Animal Anatomy Laboratory in Ecuador / Conservación de Órganos (Corazón, Encéfalo y Riñón) de Canino Mediante la Técnica de Plastinación en Silicona al Frío Realizada en Laboratorio de Anatomía Animal en Ecuador

SUMMARY: For the purposes of teaching anatomy, the use of cadaver preparations is considered the most efficient way of ensuring that students retain knowledge. Nevertheless, in Ecuador the use of animal specimens in universities must comply with the internationally accepted principles of replacement, reduction and refinement (3Rs). Plastination is an alternative technique which allows organs to be conserved in the long term and complies with the 3Rs. The object of the present work was to use cold-temperature silicone plastination with Biodur® products to obtain long-lasting, easy-to-handle canine organs for use as tools for the teaching of animal anatomy. Six canine cadavers were obtained from local animal protection charities. The hearts, brains and kidneys of the cadavers were dissected and fixed with formaldehyde 10 %. They were then dehydrated with acetone at -20 °C. The specimens were impregnated with Biodur® S10:S3 (-20 °C) and finally cured with Biodur® S6. We plastinated six hearts, twelve kidneys, four brains and one encephalic slice of canine. The application of cold-temperature plastination to canine organs followed the parameters established for the conventional protocol, enabling us to obtain organs of brilliant appearance, free of odours, in which the anatomical form was preserved. Thus the technique helped us to comply with the 3Rs, as we obtained easy-to-handle teaching models to replace fresh or formaldehyde-fixed samples for the teaching-learning of the canine anatomy.

En la enseñanza de la Anatomía, el uso de preparaciones cadavéricas se considera el método que permite a los estudiantes retener el conocimiento de una forma más eficiente. No obstante, en Ecuador, el uso de especímenes animales en las universidades se debe realizar bajo el principio internacional de reemplazo, reducción y refinamiento (3Rs). La técnica de plastinación es una técnica alternativa que permite preservar órganos a largo plazo y que se adapta al principio de las 3Rs. El objetivo del trabajo fue utilizar la técnica de plastinación en silicona al frío con productos Biodur® para obtener órganos caninos duraderos y manejables útiles como herramienta para la enseñanza de la anatomía animal. Se obtuvieron seis cadáveres de caninos de fundaciones locales para la protección animal. Se realizaron disecciones de corazones, cerebros y riñones de los cadáveres caninos. Los órganos se fijaron con formalina al 10 %. A continuación, se llevó a cabo la deshidratación con acetona a -20 °C. Los especímenes fueron impregnados con S10:S3 Biodur® (-20 °C) y al final fueron curados con Biodur® S6. Se lograron plastinar seis corazones, doce riñones, cinco encéfalos y un tallo encefálico de canino. La técnica de plastinación al frío utilizada para obtener órganos de canino conservó los parámetros empleados en el protocolo convencional y permitió obtener órganos que presentaron aspecto brillante, ausencia de olores y mantuvieron la forma anatómica. Por lo que, la técnica facilitó cumplir con el principio de las 3Rs al obtenerse modelos didácticos fáciles de manipular que pueden reemplazar muestras frescas o formolizadas en el proceso de enseñanza-aprendizaje de la anatomía del canino.

Reducción, reutilización y reciclaje: Una revisión crítica del conocimiento científico sobre las pérdidas y desperdicios de alimentos en Chile

Reducir, reutilizar y reciclar, "regla de las tres R", son estrategias que promueven una actitud responsable hacia el medioambiente y contribuyen al desarrollo sostenible. El objetivo de esta revisión fue analizar las publicaciones científicas generadas en Chile en el área de las pérdidas y desperdicios de alimentos (PDA), con enfoque en su cuantificación y basadas en "la regla de las tres R". Se realizó una búsqueda de literatura en tres bases de datos: Web of Science, Pubmed y Scopus. Los artículos originales seleccionados se clasificaron según tipo de acción, etapa de la cadena de suministro de alimentos, grupo de alimentos, cuantificación de PDA y tipo de estrategia (reducción, reutilización o reciclaje). Se encontraron 5 artículos de revisión y 31 artículos originales. La revalorización de subproductos agroindustriales fue la principal acción involucrada en los estudios de PDA (n= 22). El tipo de subproducto también fue analizado, y se encontró que los compuestos fenólicos provenientes de frutales son los más estudiados. Solo el 32% (12/31) de los artículos reportaron información sobre cuantificación de PDA. La revalorización de subproductos sumada a la recuperación de alimentos para producción de energía (n= 4) y de compostaje (n= 2) hacen que el reciclaje sea la estrategia más investigada. Estos antecedentes evidencian la necesidad de complementar la investigación nacional a nivel de acciones que apunten más hacia la reducción y reutilización de PDA y su cuantificación. Este conocimiento permitirá establecer líneas base y planes de monitoreo que contribuyan al cumplimiento de los Objetivos de Desarrollo Sostenible en Chile.

Reduce, reuse, and recycle "the 3Rs rule" are strategies that promote a responsible attitude towards the environment and contribute to sustainable development. The objective of this review was to analyze the scientific publications generated in Chile in the area of food loss and waste (FLW), with a focus on its quantification and based on "the 3Rs rule". A literature search was carried out in three databases: Web of Science, Pubmed, and Scopus. The selected original articles were classified according to the type of action, stage of the food supply chain, food group, FLW quantification, and type of strategy (reduction, reuse, or recycling). Five review articles and 31 original articles were found. The revalorization of agro-industrial by-products was the main initiative involved in the FLW studies (n= 22). The type of by-product was also analyzed, and it was found that the phenolic compounds from fruits are the most studied. Only 32% (12/31) of the articles reported information on FLW quantification. The revalorization of by-products added to food recovery for energy production (n= 4) and composting (n= 2) make recycling the most researched strategy. These antecedents show the need to complement the national research at the level of actions that point more towards reducing and reusing of FLW and its quantification. This knowledge will allow the establishment of baselines and monitoring plans that contribute to the fulfillment of the Sustainable Development Goals in Chile.

Reemplazar ideas, reducir equívocos y refinar argumentos: una reinterpretación de las 3R de la experimentación animal

Propósito/Contexto. En el presente trabajo se llevará a cabo una reinterpretación de las tres erres (3R) propuestas por William Russell y Rex Burch (reemplazo, reducción y refinamiento), con el objetivo de ampliar su alcance y mejorar las prácticas de experimentación con animales no humanos. Metodología/Enfoque. Se revisará el sentido que le dieron Russell y Burch a las 3R y se evaluará el modo en que cada una de ellas podría redefinirse o complementarse a la luz de las prácticas científicas, las posibilidades técnicas y los conocimientos bioéticos actuales vinculados al uso de animales en investigación. Resultados/Hallazgos. El artículo mostrará que 1) no solo habrían de reemplazarse animales, sino también las ideas equívocas que tenemos, tanto sobre ellos, como sobre la importancia de la educación bioética en la formación científica, 2) que la reducción, además de referirse al número de sujetos utilizados en cada experimento, debería servir para acabar con investigaciones innecesarias, repetitivas y superfluas, así como con algunos persistentes equívocos sobre el modo de operar de la ciencia y 3) que el refinamiento tendría que salir del espacio experimental para extenderse al modo en que pensamos sobre ética animal en el ámbito de la investigación. Discusión/Conclusiones/Contribuciones. El trabajo da cuenta de la importancia que tiene la incorporación del conocimiento bioético contemporáneo en las prácticas de experimentación con animales para mejorar el carácter reflexivo y ético de la ciencia.

Purpose/Background. In the present work, a reinterpretation of the 3Rs (3Rs) proposed by William Russell and Rex Burch (Replacement, Reduction and Refinement) will be carried out with the aim of broadening its scope and improving nonhuman animal experimentation practices. Methodology/Approach. The meaning given by Russell and Burch to the 3Rs will be reviewed and the way in which each of them could be redefined or complemented in the light of current scientific practices, technical possibilities and bioethical knowledge related to the use of animals in research will be evaluated. Results/Findings. The article will show that 1) not only animals should be replaced, but also the misconceptions we have, both about them and about the importance of bioethics education in scientific training, 2) that the reduction, in addition to the number of subjects used in each experiment, should serve to end unnecessary, repetitive and superfluous research, as well as some persistent misconceptions about the way science operates, and 3) that refinement should go beyond the experimental space to extend to the way we think about animal ethics in the research setting. Discussion/Conclusions/Contributions. The paper reports on the importance of incorporating contemporary bioethical knowledge into animal experimentation practices to enhance the reflexive and ethical character of science.

Objetivo/Contexto. Neste documento, uma reinterpretação dos 3Rs (3Rs) propostos por William Russell e Rex Burch (Substituição, Redução e Refinamento) será realizada com o objetivo de ampliar seu escopo e melhorar as práticas não-humanas de testes em animais. Metodologia/ Abordagem. Revisaremos o significado dado por Russell e Burch aos 3Rs e avaliaremos como cada um deles poderia ser redefinido ou complementado à luz das práticas científicas atuais, possibilidades técnicas e conhecimentos bioéticos relacionados ao uso de animais na pesquisa. Resultados/Descobertas. O artigo mostrará que 1) não somente os animais devem ser substituídos, mas também conceitos errôneos sobre eles e a importância da educação bioética no treinamento científico, 2) que a redução, além do número de sujeitos utilizados em cada experimento, deve servir para eliminar pesquisas desnecessárias, repetitivas e supérfluas, assim como alguns conceitos errôneos persistentes sobre a maneira como a ciência funciona, e 3) que o refinamento deve se estender além do espaço experimental para a maneira como pensamos sobre a ética animal na pesquisa. Discussão/Conclusões/Contribuições. O artigo explica a importância de incorporar o conhecimento bioético contemporâneo nas práticas de experimentação animal para realçar o caráter reflexivo e ético da ciência.

Regulation of calcium oscillations by IP3R and RyR in neonatal rat cardiomyocytes / 西安交通大学学报(医学版)

【Objective】 To explore the roles of 1,4,5-trisphosphate receptor (IP3Rs) and ryanodine receptors (RyRs), the two main Ca2+ release channels in sarcoplasmic reticulum (SR), in the regulation of intracellular Ca2+ oscillations in neonatal rat cardiomyocytes (NRCMs). 【Methods】 We isolated and cultured NRCMs for different days, then loaded them with Ca2+ indicator fura-2 and performed real-time fluorescent imaging. To distinguish the effects of IP3Rs and RyRs, NRCMs were pre-treated with phenylephrine (PE, IP3Rs agonist), caffeine (RyRs agonist), 2-APB (IP3Rs antagonist), and tetracaine (RyRs antagonists), respectively. 【Results】 The cultured monolayer NRCMs showed spontaneous synchronized Ca2+ oscillations. PE activation or 2-APB blockade of IP3Rs increased or reduced the frequency of Ca2+ oscillations in NRCMs, accordingly, with no significant effect on the amplitude of Ca2+ oscillations. Activation of RyRs with caffeine increased the frequency of Ca2+ oscillations, but unsynchronized the intercellular rhythm of calcium release and beating pace, while blocking RyRs with tetracaine completely abolished the Ca2+ oscillations and beats in NRCMs. In addition, the effect of PE stimulation on Ca2+ oscillation frequency gradually decreased along with cultured days. 【Conclusion】 IP3Rs regulate the rhythm of calcium oscillations, whereas RyRs are the main channel for bulky store Ca2+ release.

Initial survey on the use of animals in scientific research and teaching reveals divided opinion of the Brazilian population / Investigação inicial sobre o uso de animais na pesquisa científica e no ensino revela opinião dividida da população brasileira

ABSTRACT Objective: Specific legislation regulating the use of animals in research in Brazil was introduced in 2008. However, the viewpoint of the Brazilian population regarding the use of animals in research and teaching activities remains largely unknown. Investigation of the public viewpoint on and understanding of the topic is required given the current shifts in the animal ethics scenario in Brazil. The objective of this study was to provide the first insight into the Brazilian population viewpoint on the use of animals in scientific research and teaching activities. Methods: Data collected in a survey involving 2,115 individuals aged 16 years or older and residing in 130 municipalities distributed across the five Brazilian macroregions (North, Northeast, South, Southeast, and Midwest) were analyzed. The margin of error for entire sample was set at 2%, with a 95% confidence interval. Results: This survey revealed that most Brazilian citizens are in favor of the use animals in research, particularly for medical purposes. Different views depending on the nature of research were identified. Approximately 80% of respondents were also in favor of frequent oversight of laboratories and animal facilities. Conclusion: Survey findings indicate that the opinion of the Brazilian population is divided when it comes to the use of animals in scientific research and teaching. Divided opinions expose a limited understanding of the importance of basic sciences and emphasizes the need for improved communication between the scientific community and the general population. Further strategies aimed to promote animal welfare are discussed.

RESUMO Objetivo: A legislação específica que regula o uso de animais em pesquisa no Brasil foi introduzida em 2008. No entanto, a opinião da população brasileira sobre o uso de animais em atividades de pesquisa e ensino ainda é desconhecida. No atual cenário brasileiro em mudança com relação à ética animal, é necessário avaliar as visões e o conhecimento da população sobre o assunto. O objetivo deste destudo foi realizar o primeiro levantamento da opinião da população brasileira sobre o uso de animais em atividades de ensino e pesquisa científica. Métodos: Analisamos os resultados de uma pesquisa com 2.115 indivíduos com 16 anos ou mais de 130 municípios das cinco macrorregiões brasileiras (Norte, Nordeste, Sul, Sudeste e Centro-Oeste). A margem de erro para toda a amostra foi de 2% dentro de um intervalo de confiança de 95%. Resultados: A pesquisa revelou que a maioria da população brasileira era favorável ao uso de animais em pesquisas, principalmente para fins médicos. Diferentes pontos de vista, dependendo da natureza da pesquisa, também foram identificados. Além disso, aproximadamente 80% dos entrevistados eram favoráveis ao monitoramento frequente de laboratórios e instalações de animais. Conclusão: A opinião da população brasileira está dividida com relação ao uso de animais em pesquisa e ensino científicos. Essa divisão expõe um entendimento limitado da importância das ciências básicas e destaca a necessidade de uma melhor comunicação entre a comunidade científica e a população em geral. Outras ações para alcançar as melhorias desejadas no bem-estar animal são discutidas.

Humane endpoint in mice by Brazilian researchers in the vaccine sector / Ponto final humanitário em camundongos aplicado por pesquisadores brasileiros da área de vacinas

The application of a humane endpoint (HE) for mice in vaccine trials and further challenging tasks with lethal samples is necessary to reduce or prevent pain and suffering in these animals, and is a refinement of the 3R policy enforced for animal testing in both national and international scenarios. In order to investigate the application of HE in Brazil, researchers from the vaccine sector have answered a questionnaire about their research profile, their usage of HE and their knowledge of its defining criteria, monitoring of animals, staff training, and euthanasia methods employed. The main results revealed that researchers failed to recognise the very concept of HE as well as when to apply it. In addition, the Institutional Animal Care and Use Committees (IACUCs) failed upon giving their approval to the trials. HE is an important refinament of animal testing policy, and these results highlight the need for a clear pre-established definition of when and how it should be implemented in order to ensure more effective application. Furthermore, it is important to clarify the ethics involved and the commitment of both the research teams and IACUCs to animal welfare.(AU)

A aplicação de um ponto final humanitário (PFH) em experimentos com vacinação e posterior desafio com amostras letais em camundongos é necessária para diminuir ou impedir a dor e o sofrimento desses animais, constituindo refinamento de acordo com a política dos 3Rs, vigente em âmbito nacional e internacional, no cenário da experimentação animal. A fim de investigar a aplicação de PFH no Brasil, foi enviado um questionário para pesquisadores da área de vacinas, com perguntas sobre perfil do pesquisador, aplicação de PFH e critérios para sua definição, monitoramento dos animais, treinamento da equipe e métodos de eutanásia empregados. Os principais resultados revelaram que há falhas tanto em reconhecer o conceito quanto em aplicar o PFH, por parte dos pesquisadores, e que as CEUAs também falharam ao aprovar tais experimentos. Chama-se atenção para o fato de que o PFH é um importante refinamento e para a necessidade de definição prévia de PFH e sua aplicação, assim como para a importância do esclarecimento da questão ética envolvida e do necessário compromisso das equipes de pesquisa e das CEUAs com o bem-estar animal.(AU)

Chemical Constituents of Chimonanthus nitens Oliv / 中国药学杂志

OBJECTIVE: To study the chemical constituents from the leaves of Jiangxi genuine medicinal material Chimonanthus nitens Oliv. METHODS: The compounds were isolated and purified by silica gel column chromatography, Sephadex LH-20, ODS column chromatography, semi-preparative HPLC, and so on. Their structures were elucidated on the basis of physiochemical properties and spectral data. RESULTS: Ten compounds were isolated and elucidated as uracil (1), 6, 7-dimethoxycoumarin (2), 6, 7, 8-trimethoxycoumarin (3), p-hydroxybenzoic acid ethyl ester (4), 1∶1 mixture of two diastereomers identical with (3RS, 6RS)-2, 6-dimethyl-octa-1, 7-dien-3, 6-diol (5 and 6), kaempferol (7), isofraxidin (8), scopoletin (9), and loliolide (10). CONCLUSION: Compounds 1, 3-6 and 10 are isolated from this plant for the first time and compounds 1, 4-6 are isolated from this genus for the first time.

PNPLA3 rs738409 Polymorphism Associated with Hepatic Steatosis and Advanced Fibrosis in Patients with Chronic Hepatitis C Virus: A Meta-Analysis

BACKGROUND/AIMS: The recognition of a correlation between patatin-like phospholipase domain containing-protein 3 (PNPLA3) rs738409 (C>G) and the severity of liver steatosis or fibrosis in chronic hepatitis C (CHC) has not reached a consensus. This meta-analysis sought to investigate with accuracy the association between the PNPLA3 rs738409 (C>G) polymorphism and liver steatosis and advanced fibrosis in CHC patients. METHODS: We performed a comprehensive literature search from the PubMed, Embase, Web of Science, and Google Scholar databases up to December 31, 2014. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were calculated. Statistical analyses were performed using Stata 12.0 software. RESULTS: The meta-analysis revealed the severity of liver fibrosis was significantly higher in CHC patients with PNPLA3 rs738409 GG in Caucasians (versus CC+CG: OR, 2.29; 95% CI, 1.57 to 3.35; pG) was associated with the risk of both advanced liver fibrosis and steatosis in patients with CHC, especially among Caucasian populations.

How can standardised reporting of animal research advance the 3 Rs? / 中国实验动物学报

Although the publication of peer-reviewed scientific papers is the major way in which scientists communicate with one another, these papers often lack essential information about the conditions under which the animals lived and how they were treated during the experiments.Without this information it may be impossible to evaluate the studies , replicate them in another laboratory , or use them to advance the 3Rs in other animal experiments .This paper gives some advice on how the reporting of animal research can be improved .

Ethical Guidelines for Use of Experimental Animals in Biomedical Research

Increasing concerns on animal welfare, the discussion about the necessity and the ethical acceptability of animal experiments is getting important. In this article, we describe ethical issues between the benefits of the animal experiments and the suffering of the animals. If there are no alternatives for animal experiments, the ethical guidelines, 3Rs (Reduction, Replacement, Refinement) by Russell and Burch for humane animal experiments should be considered. Moreover, Institutional Animal Care and Use Committee (IACUC) and Institutional Biosafety Committee (IBC) assist researchers to perform animal testing humanely and safely. We point out three important aspects in support of humane animal experiments; 1) the development of diverse scientific alternatives on the animal testing, 2) the ethical education of animal experiments for researchers, 3) the productive interactions among various social groups on ethics and safety of animal experiments.

Improved assessment of frozen/thawed mouse spermatozoa using fluorescence microscopy

Genetically modified (GM) animals are unique mutants with an enormous scientific potential. Cryopreservation of pre-implantation embryos or spermatozoa is a common approach for protecting these lines from being lost or to store them in a repository. A mutant line can be taken out of a breeding nucleus only if sufficient numbers of samples with an appropriate level of quality are cryopreserved. The quality of different donors within the same mouse line might be heterogeneous and the cryopreservation procedure might also be error-prone. However, only limited amounts of material are available for analysis. To improve the monitoring of frozen/thawed spermatozoa, commonly used in vitro fertilization (IVF) followed by embryo transfer were replaced with animal-free techniques. Major factors for assessing spermatozoa quality (i.e., density, viability, motility, and morphology) were evaluated by fluorescence microscopy. For this, a live/dead cell staining protocol requiring only small amounts of material was created. Membrane integrity was then examined as major parameter closely correlated with successful IVF. These complex analyses allow us to monitor frozen/thawed spermatozoa from GM mice using a relatively simple staining procedure. This approach leads to a reduction of animal experiments and contributes to the 3R principles (replacement, reduction and refinement of animal experiments).