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.

Mechanism of Yanghe Decoction against subcutaneous tumor in pulmonary metastasis from breast cancer through HIF-1α signaling pathway regulating glycolysis:based on network pharmacology and animal experiment / 中国中药杂志

This study aims to explore the mechanism of Yanghe Decoction(YHD) against subcutaneous tumor in pulmonary metastasis from breast cancer, which is expected to lay a basis for the treatment of breast carcinoma with YHD. The chemical components of medicinals in YHD, and the targets of the components were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and SwissTargetPrediction. The disease-related targets were searched from GeneCards and Online Mendelian Inheritance in Man(OMIM). Excel was employed to screen the common targets and plot the Venn diagram. The protein-protein interaction network was constructed. R language was used for Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment. A total of 53 female SPF Bablc/6 mice were randomized into normal group(same volume of normal saline, ig), model group(same volume of normal saline, ig), and low-dose and high-dose YHD groups(YHD, ig, 30 days), with 8 mice in normal group and 15 mice in each of the other groups. Body weight and tumor size was measured every day. Curves for body weight variation and growth of tumor in situ were plotted. In the end, the subcutaneous tumor sample was collected and observed based on hematoxylin and eosin(HE) staining. The mRNA and protein levels of hypoxia inducible factor-1α(HIF-1α), pyruvate kinase M2(PKM2), lactate dehydrogenase A(LDHA), and glucose transporter type 1(GLUT1) were detected by PCR and Western blot. A total of 213 active components of YHD and 185 targets against the disease were screened out. The hypothesis that YHD may regulate glycolysis through HIF-1α signaling pathway to intervene in breast cancer was proposed. Animal experiment confirmed that the mRNA and protein levels of HIF-1α, PKM2, LDHA, and GLUT1 in the high-and low-dose YHD groups were lower than those in the model group. YHD has certain inhibitory effect on subcutaneous tumor in pulmonary metastasis from breast cancer in the early stage, which may intervene pulmonary metastasis from breast cancer by regulating glycolysis through HIF-1α signaling pathway.

Animal experimental study on the effects of different levels of amputation on cardiovascular system / 生物医学工程学杂志

Vascular injury resulting from lower limb amputation leads to the redistribution of blood flow and changes in vascular terminal resistance, which can affect the cardiovascular system. However, there was no clear understanding of how different amputation levels affect the cardiovascular system in animal experiments. Therefore, this study established two animal models of above-knee amputation (AKA) and below-knee amputation (BKA) to explore the effects of different amputation levels on the cardiovascular system through blood and histopathological examinations. The results showed that amputation caused pathological changes in the cardiovascular system of animals, including endothelial injury, inflammation, and angiosclerosis. The degree of cardiovascular injury was higher in the AKA group than in the BKA group. This study sheds light on the internal mechanisms of amputation's impact on the cardiovascular system. Based on the amputation level of patients, the findings recommend more comprehensive and targeted monitoring after surgery and necessary interventions to prevent cardiovascular diseases.

Exploring the treatment of sepsis-associated acute lung injury with Liangge Powder via ERK1/2 and PI3K/AKT pathways: based on network pharmacology and whole animal experimentation / 中华劳动卫生职业病杂志

Objective: To investigate the therapeutic effect and mechanism of Liangge Powder against sepsis-induced acute lung injury (ALI) . Methods: From April to December 2021, the key components of Liangge Powder and its targets against sepsis-induced ALI were analyzed by network pharmacology, and to enrich for relevant signaling pathways. A total of 90 male Sprague-Dawley rats were randomly assigned to sham-operated group, sepsis-induced ALI model group (model group), Liangge Powder low, medium and high dose group, ten rats in the sham-operated group and 20 rats in each of the remaining four groups. Sepsis-induced ALI model was established by cecal ligation and puncture. Sham-operated group: gavage with 2 ml saline and no surgical treatment. Model group: surgery was performed and 2 ml saline was gavaged. Liangge Powder low, medium and high dose groups: surgery and gavage of Liangge Powder 3.9, 7.8 and 15.6 g/kg, respectively. To measure the wet/dry mass ratio of rats lung tissue and evaluate the permeability of alveolar capillary barrier. Lung tissue were stained with hematoxylin and eosin for histomorphological analysis. The levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL) -6 and IL-1β in bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay. The relative protein expression levels of p-phosphatidylinositol 3-kinase (PI3K), p-protein kinase B (AKT), and p-ertracellular regulated protein kinases (ERK) were detected via Western blot analysis. Results: Network pharmacology analysis indicated that 177 active compounds of Liangge Powder were selected. A total of 88 potential targets of Liangge Powder on sepsis-induced ALI were identified. 354 GO terms of Liangge Powder on sepsis-induced ALI and 108 pathways were identified using GO and KEGG analysis. PI3K/AKT signaling pathway was recognized to play an important role for Liangge Powder against sepsis-induced ALI. Compared with the sham-operated group, the lung tissue wet/dry weight ratio of rats in the model group (6.35±0.95) was increased (P<0.001). HE staining showed the destruction of normal structure of lung tissue. The levels of IL-6 [ (392.36±66.83) pg/ml], IL-1β [ (137.11±26.83) pg/ml] and TNF-α [ (238.34±59.36) pg/ml] were increased in the BALF (P<0.001, =0.001, <0.001), and the expression levels of p-PI3K, p-AKT and p-ERK1/2 proteins (1.04±0.15, 0.51±0.04, 2.31±0.41) were increased in lung tissue (P=0.002, 0.003, 0.005). The lung histopathological changes were reduced in each dose group of Liangge Powder compared with the model group. Compared with the model group, the wet/dry weight ratio of lung tissue (4.29±1.26) was reduced in the Liangge Powder medium dose group (P=0.019). TNF-α level [ (147.85±39.05) pg/ml] was reduced (P=0.022), and the relative protein expression levels of p-PI3K (0.37±0.18) and p-ERK1/2 (1.36±0.07) were reduced (P=0.008, 0.017). The wet/dry weight ratio of lung tissue (4.16±0.66) was reduced in the high-dose group (P=0.003). Levels of IL-6, IL-1β and TNF-α[ (187.98±53.28) pg/ml, (92.45±25.39) pg/ml, (129.77±55.94) pg/ml] were reduced (P=0.001, 0.027, 0.018), and relative protein expression levels of p-PI3K, p-AKT and p-ERK1/2 (0.65±0.05, 0.31±0.08, 1.30±0.12) were reduced (P=0.013, 0.018, 0.015) . Conclusion: Liangge Powder has therapeutic effects in rats with sepsis-induced ALI, and the mechanism may be related to the inhibition of ERK1/2 and PI3K/AKT pathway activation in lung tissue.

Inovação e ética: a adoção de técnicas substitutivas ao uso de animais no diagnóstico laboratorial da raiva / Innovation and ethics; the adoption of alternative techniques to the use of animals in the laboratory diagnosis of rabies

Abordagens interdisciplinares vêm ganhando maior reconhecimento e destaque nas comunidades de saúde humana e animal, principalmente pela (re)emergência de diversas doenças infecciosas que emanam da interface humano-animal-ambiente. A raiva, zoonose grave, considerada endêmica no Brasil e globalmente negligenciada, é um exemplo. Tanto a vigilância epidemiológica quanto a confirmação dessa doença dependem do diagnóstico laboratorial, que é realizado, frequentemente, por meio dos testes de Imunofluorescência Direta (IFD) e de Isolamento Viral em Camundongo (IVC), via inoculação intracerebral da amostra suspeita em camundongos lactentes ou desmamados. Entretanto, recentemente, a Organização Mundial da Saúde reconheceu a Transcrição Reversa seguida da Reação em Cadeia da Polimerase (RT-PCR) como uma técnica primária válida para esse diagnóstico, podendo ser empregada como alternativa ao uso de animais, evitando sofrimento e morte. Esta dissertação apresenta uma discussão sobre as implicações técnicas e éticas da (não) adoção desse método substitutivo, considerando que todos os animais devem ser respeitados e entendidos como sujeitos singulares em suas percepções do mundo, não como objetos de pesquisa. Esse fato corrobora a necessidade de novas perspectivas que ressignifiquem nossas relações com os animais não humanos, o que é primordial para o estabelecimento de mudanças sistêmicas, de caráter ético-político, que visem o fim da instrumentalização animal e de seu uso no âmbito científico, bem como de qualquer forma de opressão.

Interdisciplinary approaches have been gaining greater recognition and prominence in the human and animal health communities, mainly due to the (re)emergence of several infectious diseases that emanate from the human-animal-environment interface. Rabies is an example, considered a serious zoonosis endemic in Brazil and globally neglected. Both epidemiological surveillance and confirmation of this disease depend on laboratory diagnosis, which is usually performed by the direct fluorescent antibody test (DFAT) and the mouse inoculation test (MIT) via intracranial inoculation of the suspected sample into suckling or weanling mice. However, the World Health Organization recently recognized the reverse transcription polymerase chain reaction (RT-PCR) as a valid primary technique for this diagnosis, which can replace the use of animals, avoiding suffering and death. This study presents a discussion about the technical and ethical implications of (not) adopting this alternative method, considering that all animals must be respected and understood as unique beings with their perceptions of the world, not as objects of research. It also further corroborates the need for new perspectives that reframe our relationships with non-human animals, which is fundamental for the implementation of systemic ethical-political changes, aiming at the end of animal instrumentalization and use in scientific research, as well as all forms of oppression.

Consideration on Animal Experiment in PET/CT / 中国医疗器械杂志

PET/CT imaging can reflect the physiological metabolic process in living body which is the model experiment incapable to simulate. Animal experiment may be considered for systematic validation of PET/CT products. The obtained research data can be used to evaluate the feasibility, effectiveness and safety of PET/CT products, and be submitted as supporting documents for research data or clinical evaluation data when doing product registration or alteration registration. In this study, the functions and advantages of animal experiments were expounded, and relevant research cases were given as well as the issues that should be paid attention to. It can be a reference for the validation and review of PET/CT products.

Effect and mechanism of Jingqi Yukui Capsules on gastric ulcer mucosa healing quality: based on network pharmacology and animal experiment / 中国中药杂志

This study aims to identify the active components and the mechanism of Jingqi Yukui Capsules(JQYK) in the treatment of gastric ulcer based on network pharmacology, and verify some key targets and signaling pathways through animal experiment. To be specific, first, the active components and targets of JQYK were retrieved from a Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine(BATMAN-TCM) and Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and the targets of gastric ulcer from GeneCards and Online Mendelian Inheritance in Man(OMIM) with the search term "gastric ulcer". The common targets of the two were the potential targets of the prescription for the treatment of the di-sease. Then, protein-protein interaction(PPI) network of key targets were constructed based on STRING and Cytoscape 3.7.2, followed by Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment by matescape database and pathway visualization by Omicshare. For the animal experiment, the improved method of Okabe was used to induce gastric ulcer in rats, and the model rats were classified into the model group, JQYK high-dose(JQYK-H), medium-dose(JQYK-M), and low-dose(JQYK-L) groups, Anweiyang Capsules(WYA) group, and Rabeprazole Sodium Enteric Capsules(RBPZ) group. Normal rats were included in the blank group. Rats in the blank group and model group were given distilled water and those in the administration groups received corresponding drugs. Then gastric ulcer healing in rats was observed. The changes of the gastric histomorphology in rats were evaluated based on hematoxylin-eosin(HE) staining, and the content of inducible nitric oxide synthase(iNOS) in rat gastric tissue was detected with Coomassie brilliant blue method. The mRNA and protein levels of some proteins in rat gastric tissue were determined by real-time quantitative polymerase chain reaction(RT-qPCR) and Western blot(WB) to further validate some key targets and signaling pathways. A total of 206 active components and 535 targets of JQYK, 1 305 targets of gastric ulcer, and 166 common targets of the disease and the drug were yielded. According to PPI analysis and KEGG pathway enrichment analysis, multiple key targets, such as interleukin-6(IL-6), tumor necrosis factor(TNF), mitogen-activated protein kinase 1(MAPK1), MAPK3, and MAPK14, as well as nuclear factor kappa-B(NF-κB) signaling pathway, IL-17 signaling pathway, and leukocyte transendothelial migration in the top 20 key signaling pathways were closely related to inflammation. The key protein p38 MAPK and NF-κB signaling pathway were selected for further verification by animal experiment. The gastric ulcer in the JQYK-H group recovered nearly to the level in the blank group, with significant decrease in the content of iNOS in rat gastric tissue and significant reduction in the mRNA and phosphorylation levels of p38 MAPK and the mRNA and protein levels of NF-κB p65 in rat gastric tissue. The results indicated that JQYK can inhibit the phosphorylation of the key protein p38 MAPK and the expression of NF-κB p65 in the NF-κB signaling pathway, thereby exerting the anti-inflammatory effect and effectively improving the quality of gastric ulcer healing in rats. Thus, the animal experiment result verifies some predictions of network pharmacology.

Mechanism of Carthami Flos and Lepidii Semen drug pair in inhibition of myocardial fibrosis by improving cardiac microenvironment based on network pharmacology and animal experiment / 中国中药杂志

Previously, Carthami Flos and Lepidii Semen(CF-LS) drug pair has been proved effective in inhibiting myocardial fibrosis(MF) by blunting the activity of cardiac fibroblasts. The present study explored the underlying mechanism of CF-LS in inhibiting MF by improving the cardiac microenvironment based on network pharmacology combined with experimental verification. Active compounds and potential targets of CF-LS were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and the potential targets of MF were obtained from GeneCards, Online Mendelian Inheritance in Man(OMIM), and Pharmacogenetics and Pharmacogenomics Knowledge Base(PharmGKB). The "active component-target-MF" network was constructed and analyzed by Cytoscape 3.8.1. The protein-protein interaction(PPI) network was constructed by STRING. The Gene Ontology(GO) biological process enrichment analysis was performed by CluoGO plug-in. Kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathway enrichment analysis was performed by R 4.0.2 and Funrich. Subsequently, the inhibitory effect of CF-LS on MF was investigated based on angiotensin Ⅱ(Ang Ⅱ)-induced MF rats. RT-PCR and ELISA were used to verify the effect of CF-LS on the targets of signaling pathways related to vascular endothelial cells predicted by the network pharmacology. Thirty-one active components and 204 potential targets of CF-LS, 4 671 MF-related targets, and 174 common targets were obtained. The network analysis showed that the key targets of CF-LS against MF included RAC-alpha serine/threonine-protein kinase(AKT1), transcription factor AP-1(JUN), mitogen-activated protein kinase 1(MAPK1), cellular tumor antigen p53(TP53), transcription factor p65(RELA), and mitogen-activated protein kinase 8(MAPK8). Biological processes mainly involved regulation of blood vessel diameter, regulation of blood vessel endothelial cell migration, cell death in response to oxidative stress, etc. Advanced glycation end products(AGE)-receptor for advanced glycation end products(RAGE) signaling pathway, phosphoinositide 3-kinase(PI3 K)-serine/threonine protein kinase(AKT) signaling pathway, hypoxia-inducible factor-1(HIF-1) signaling pathway, integrin signaling pathway, transforming growth factor-β(TGF-β) signaling pathway, etc. were involved in signaling pathway enrichment. Literature retrieval confirmed that some of these signaling pathways were closely related to vascular endothelial cells, including AGE-RAGE, PI3 K-AKT, HIF-1α, p53, the transcription factor activator protein-1(AP-1), integrin, p38 MAPK, and TGF-β. Animal experiments showed that CF-LS inhibited MF induced by Ang Ⅱ in rats by suppressing the expression of RAGE, HIF-1α, integrin β6, and TGF-β1. The inhibitory effect of CF-LS on MF has the characteristics of multiple components, multiple targets, and multiple pathways. CF-LS can inhibit MF by regulating the activity of vascular endothelial cells in the cardiac microenvironment.

La relevancia moral del dolor de animales de experimentación y de producción / The moral relevance of pain in experimental and production animals

RESUMEN: El aspecto emocional del dolor no solo es controvertido en aves, peces y roedores, sino también en los seres humanos autoconscientes. Cuando el dolor emocional, o la insatisfacción, es vista como un mal intrínseco, evitarla o, al menos, minimizarla es un deber moral directo. El objetivo de este artículo es mostrar que no solo los modelos animales mamíferos que se utilizan en experimentación -rata (Rattus norvegicus) y ratón (Mus musculus)- sienten dolor, como reconocen universalmente las instituciones que se ocupan del bienestar animal, sino también los peces de laboratorio (Danio rerio) y los de producción (Salmo salar), y que este hecho es moralmente relevante desde el punto de vista de todas las concepciones de la moral que suelen enfrentarse en los debates morales públicos y académicos. En consecuencia, también en el caso de los peces habría que tener en cuenta indicadores de bienestar animal.

SUMMARY: The emotional aspect of pain is not only controversial in fish and rodents, but also in self-conscious humans. When pain or dissatisfaction is seen as an intrinsic evil, avoiding it or at least minimizing it is a moral duty direct. The objective of this article is to refer to pain, stress and the most widely used indices of animal welfare both in laboratory fish (Danio rerio) and production fish (Salmo salar) as well as in rat experimental animal models (Rattus norvegicus) and mouse (Mus musculus).

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.

Baseline growth of the Trinidad freshwater crab Poppiana dentata (Randall, 1840) under laboratory conditions / Crescimento do caranguejo de água doce de Trinidad, Poppiana dentata (Randall, 1840), em condições laboratoriais

Poppiana dentata (Randall, 1840) is widely distributed throughout riverine habitats in Trinidad. However, there is a scarcity of information on the biology of this species. This study provides the first baseline examination that describes growth aspects for P. dentata. Juvenile crabs were obtained from berried females collected in northwest Trinidad. Carapace width (CW), length (CL), moult incident, intermoult period and qualitative aspects were recorded for crabs (N = 23) over 9 months. CW, CL and intermoult period were used to derive percentage size increment, specific growth rate (SGR) and size at structural maturity for both sexes. Growth curves and logistic equations were also generated for each sex. Hatched crabs (< 5 mm CW) underwent rapid hardening after their first moult, indicating a fast turnover of moult cycles. SGR and CW percent increment were also the highest for this initial moult (P < 0.05). CW, CL, intermoult period, size increment and SGR did not differ between sexes (P > 0.05), with logistic equations expressed as CW = 32.81 (1+exp (1.481 ̵ 0.031t))-1 for males and CW = 34.07 (1+exp (1.516 ̵ 0.027t))-1 for females. Yet, breakpoint analyses indicated dissimilar sizes for structural maturity (male: 28.40 mm CW; female: 16.84 mm CW). These patterns reflect a shorter life span for this species in comparison to what has been reported for other trichodactylid relatives. This can have implications for P. dentata populations residing in anthropogenically disturbed habitats; thus, highlighting the need for conservation strategies to ensure preservation of native wild stock.

O Poppiana dentata (Randall, 1840) está amplamente distribuído nos habitats fluviais de Trinidad. Existem, porém, poucas informações sobre a biologia dessa espécie. Este estudo fornece a primeira análise de referência que descreve aspectos do crescimento do P. dentata. Caranguejos jovens foram obtidos a partir de fêmeas em desova na região noroeste de Trinidad. A largura da carapaça (LC), o comprimento da carapaça (CC), a incidência de muda, o período de intermuda e aspectos qualitativos foram registrados para caranguejos (N = 23) ao longo de mais de nove meses. A LC, o CC e o período de intermuda foram utilizados para obter o aumento porcentual, a taxa específica de crescimento (TEC), e o tamanho na maturidade estrutural para os dois sexos. Curvas de crescimento e equações logísticas foram também geradas para cada sexo. Os caranguejos que nasceram (com largura de carapaça < 5 mm) apresentaram um endurecimento rápido depois de sua primeira muda, o que indica uma rápida rotação dos ciclos de muda. A TEC e o aumento da porcentual da LC foram também os mais altos para esta muda inicial (P < 0,05). Não houve variação da LC, CC, do período de intermuda, aumento do tamanho e da TEC entre os sexos (P > 0,05), e as equações logísticas foram expressas como: LC = 32,81 (1+exp (1,481 ̵ 0,031t))-1 para machos, e LC = 34,07 (1+exp (1,516 ̵ 0,027t))-1 para fêmeas. Porém, as análises de quebra indicaram tamanhos diferentes para maturidade estrutural (LC para macho: 28,40; para fêmea: 16,84 mm). Esses padrões refletem um período de vida mais curto para esta espécie em comparação com o que tem sido registrado para outras famílias de Trichodactylidae. Isso pode ter implicações para as populações de P. dentata que residem em habitats modificados antropogenicamente; destaca-se, assim, a necessidade de estratégias de conservação para assegurar a preservação das populações selvagens nativas.

Inovação biomédica e ética: técnicas substitutivas na experimentação animal / Biomedical innovation and ethics: alternatives to animal testing / Innovación biomédica y ética: técnicas alternativas a la experimentación animal

Resumo A utilização de animais não humanos como ferramenta de pesquisa biomédica e em testes da indústria para consumo humano foi incorporada às práticas científicas e assimilada como fundamental. A revisão sistemática dos resultados de protocolos de fases pré-clínicas não é prática corrente, mas metanálises recentes questionam a capacidade de projeção desses dados para a espécie humana. Atualmente, junto com o questionamento científico há abrangente discussão ética sobre os conflitos inerentes à instrumentalização da vida do animal não humano, cujo ápice é alcançado na criação de animais transgênicos. O objetivo deste artigo é discutir a aplicação do conceito de vulnerabilidade ao animal não humano no contexto da experimentação e pensar as relações de poder implícitas nessas práticas. Como aplicação prática da teoria exposta, propõe-se implantar e desenvolver técnicas substitutivas ao modelo animal, que conjuguem ética e inovação.

Abstract The use of non-human animals has been incorporated into scientific practices as an essential biomedical research tool and in industry tests for human consumption. The systematic review of protocol results of preclinical phases is not a common practice, but recent meta-analyses question the projection accuracy of these data for humans. Currently, along with scientific questioning, there is a comprehensive ethical discussion about the conflicts in the instrumentalization of non-human life, which reached its peak with the creation of transgenic animals. This article discusses the application of the concept of vulnerability to non-human animals in experiments and reflects on the implicit power relations of these practices. We propose to implement and develop alternative techniques to the animal model, combining ethics and innovation.

Resumen El uso de animales no humanos como herramienta para la investigación biomédica y en pruebas de la industria para el consumo humano se ha incorporado a las prácticas científicas y se ha asimilado como fundamental. La revisión sistemática de los resultados de protocolos de fases preclínicas no es una práctica corriente, pero metaanálisis recientes cuestionan la capacidad proyección de estos datos a la especie humana. Actualmente, junto con el cuestionamiento científico, hay una discusión ética sobre los conflictos inherentes a la instrumentalización de la vida del animal no humano, que alcanza su ápice en la creación de animales transgénicos. Este artículo tiene como objetivo discutir la aplicación del concepto de vulnerabilidad al animal no humano en el contexto de la experimentación y proponer una reflexión sobre las relaciones de poder implícitas en estas prácticas. Como una aplicación práctica de la teoría expuesta, se propone implantar y desarrollar técnicas alternativas al modelo animal, que conjuguen ética e innovación.

Efeito do extrato das folhas da Passiflora edulis na cicatrização da pele em ratos / Efecto del extracto de hoja de Passiflora edulis en la cicatrización de la piel en ratas / Effect of Passiflora edulis leaf extract on skin healing in rats

Objetivo: avaliar o efeito do extrato das folhas da Passiflora edulis na cicatrização da pele em ratos, especificamente na área da lesão e na proliferação de fibroblastos. Materiais e método: estudo experimental realizado com 54 ratos da linhagem Wistar, machos, adultos, divididos igualmente em três grupos. A indução das lesões em formato circular na região dorso-cervical foi realizada com o auxílio de um punch trepano de 8 mm. As lesões do grupo de controle foram higienizadas diariamente com água filtrada e sabonete vegetal glicerinado de pH neutro. Os outros dois grupos, além da higienização diária, receberam tratamento com creme contendo extrato a 20 % das folhas da Passiflora edulis, e creme base sem extrato a 20 % das folhas da Passiflora edulis, respectivamente. No 1o, 4o, 7o e 14o dia de tratamento foram obtidas imagens para cálculo da área de lesão. No 4o, 7o e 14o dia de tratamento foram coletados espécimes para análises histológicas. Resultados: não foram encontradas diferenças estatisticamente significativas entre os grupos experimentais com relação à comparação de médias da área de lesão e de proliferação de fibroblastos. Conclusões: o uso tópico do extrato a 20 % das folhas da Passiflora edulis não apresentou efeito na área de lesão e na proliferação de fibroblastos na cicatrização da pele em ratos.

Objetivo: evaluar el efecto del extracto de hoja de Passiflora edulis en la curación de la piel en ratas, específicamente el área de la lesión y de la proliferación de fibroblastos. Materiales y método: estudio experimental con 54 ratas Wistar macho adultas, divididas por igual en tres grupos. La inducción de lesiones de forma circular en la región dorsocervical se realizó con la ayuda de un punzón de 8 mm. Las lesiones del grupo de control se limpiaron diariamente con agua filtrada y jabón vegetal glicerinado con pH neutro. Los otros dos grupos, además de la limpieza diaria, recibieron tratamiento con crema que contenía extracto al 20 % de las hojas de Passiflora edulis, y crema base sin extracto al 20 % de las hojas de Passiflora edulis, respectivamente. Los días 1, 4, 7 y 14 de tratamiento, se obtuvieron imágenes para calcular el área de la lesión. Los días 4, 7 y 14 de tratamiento, se recolectaron muestras para análisis histológicos. Resultados: no se encontraron diferencias estadísticamente significativas entre los grupos experimentales con respecto a la comparación de las medias del área de la lesión y la proliferación de fibroblastos. Conclusiones: el uso tópico del extracto al 20 % de las hojas de Passiflora edulis no tuvo efecto en el área de la lesión y de la proliferación de fibroblastos en la curación de la piel en ratas.

Objective: To evaluate the effect of Passiflora edulis leaf extract on skin healing in rats, specially the lesion area and on fibroblast proliferation. Materials and method: Experimental study with 54 male adult Wistar rats equally divided into three groups. The induction of circular-shaped lesions in the dorso cervical region was performed with the aid of an 8 mm climbing punch. Control group lesions were daily cleaned with filtered water and neutral pH glycerinated vegetable soap. The other two groups, in addition to daily cleansing, received treatment with cream containing 20 % extract of Passiflora edulis leaves and base cream without this extract, respectively. On day 1, 4, 7 and 14 of treatment, images were obtained to calculate the lesion area. On day 4, 7 and 14 of treatment, specimens were collected for histological analysis. Results: No statistically significant differences were found between the experimental groups regarding the comparison of means of lesion area and fibroblast proliferation. Conclusion: Topical use of the 20 % extract of Passiflora edulis leaves had no effect on the affected area and the proliferation of fibroblasts on skin healing in rats.

Orthotopic uterus transplantation. Sheep model experiment, Cali (Colombia) / Trasplante ortotópico de útero. Experimento en modelo ovino, Cali (Colombia)

RESUMEN Objetivo: se ha recomendado a los profesionales que tengan como proyecto realizar trasplante uterino, hacer previamente trabajos experimentales en animales. Este trabajo describe el procedimiento del trasplante uterino en ovejas y los resultados a corto y mediano plazo. Materiales y métodos: estudio de cirugía experimental en ovejas sometidas a explante y trasplante uterino. A cuatro ovejas de 40-50 kg de peso les fue trasplantado el útero (ortotópico) de cuatro ovejas vivas donantes. Se utilizó técnica de anastomosis vascular término-lateral, la vagina se suturó en un plano y el útero se fijó a la pared pélvica. Se describen las complicaciones y la evolución a 180 días Resultados: se realizó la cirugía de trasplante en las cuatro ovejas. El tiempo quirúrgico fue de 240 minutos (min) en el primer procedimiento y de 185 min en el último, y el tiempo de isquemia caliente se redujo de 42 a 22 min. Una oveja murió al séptimo día posoperatorio por una complicación intraoperatoria ajena a las anastomosis vasculares. Otra oveja que fue trasplantada desarrolló infección-local vaginal manejada con metronidazol, con evolución satisfactoria. Las tres ovejas no habían presentado rechazo al trasplante a los 6 meses. Conclusiones: el modelo ovino permite entrenamiento quirúrgico en cirugía experimental de trasplante uterino. Para los autores constituyó adquisición de conocimiento y avanzar hacia la realización futura del trasplante uterino en mujeres con Infertilidad absoluta por factor uterino en Colombia.

ABSTRACT Objective: It has been recommended that professionals who are planning to perform uterine transplantation should first carry out animal experiments. This paper describes the procedure for uterine transplant in sheep, as well as short and medium-term results. Materials and methods: Experimental surgery study in sheep subjected to uterine explantation and transplant. Four 40-50 kg sheep received uteri transplantation (orthotopic) from four live donors. End-to-side vascular anastomosis was used, the vagina was sutured on one plane and the uterus was fixed to the pelvic wall. Complications and 180-day evolution are described. Results: Transplant surgery was accomplished in the 4 sheep. Surgical time in the first procedure was 240 minutes, while the last procedure lasted 185 minutes. Warm ischemia time was reduced from 42 to 22 minutes. One sheep died on the seventh postoperative day due to an intraoperative complication unrelated to the vascular anastomosis. A second sheep developed local vaginal infection treated with metronidazole and evolved satisfactorily. No transplant rejection had occurred in the remaining 3 sheep after 6 months. Conclusions: The ovine model allowed surgical training in experimental uterine transplant surgery. For the authors, it offered an opportunity to gain knowledge and make progress towards future uterus transplantation in women with uterine factor infertility in Colombia.

Reparo inicial de defeito ósseo crítico na calvária de rato após aplicação de ondas vibratórias / Initial repair of critical bone defect in rat calvary after the application of vibratory waves

Introdução: fraturas ósseas extensas representam grande causa de morbidade e geram custos para o serviço de saúde. A vibração de baixa magnitude e alta frequência foi proposta como um tratamento alternativo para aumentar a massa óssea. Objetivo: Avaliar histomorfologicamente o reparo inicial de defeitos ósseos críticos após aplicação de ondas mecânicas vibratórias Metodologia: foram utilizados 10 Rattus norvegicus. Confeccionou-se defeitos críticos de 8,5 mm de diâmetro na calvária dos ratos. Os animais foram distribuídos em dois grupos: Grupo Controle de Defeito Ósseo (GCDO) e Grupo Experimental de Vibração Imediata (GEVI). Animais do GEVI foram submetidos a ondas vibratórias de 60 Hz e aceleração vertical de 0,3 g; elas foram aplicadas três vezes/ semana, durante vinte minutos. Após quinze dias do ato operatório, os animais foram eutanasiados para a mensuração da extensão do defeito. Considerando que estes defeitos tinham o mesmo diâmetro inicial, admitiu-se como indicador indireto de deposição osteóide, a redução da extensão linear final dos mesmos. Resultados: observou-se neoformação de matriz osteoide, restrita às bordas ósseas, em ambos os grupos. A média de extensão linear, em milímetros, do defeito ósseo do GEVI foi de 5,83 (DP=0,79) e no GCDO, foi de 6,62 (DP= 0,63). Não houve diferença estatisticamente significante entre as médias (U=8,00, z=-1,604, p=0,132). Conclusão: evidenciou-se resposta osteogênica a partir da utilização da terapêutica vibratória, contudo de forma estatisticamente não-significante. Deste modo, o presente estudo demonstrou que a utilização das ondas vibratórias não favoreceu um reparo ósseo estatisticamente significante, no período e regime vibratório estudados.

Introduction: extensive bone fractures represent a major cause of morbidity and generate costs for the health service. Vibration of low magnitude and high frequency has been proposed as an alternative treatment to increase bone mass. Objective: to evaluate histomorphologically the initial repair of critical bone defects after application of vibrating mechanical waves. Methodology: 10 Rattus norvegicus were used. Critical defects of 8.5 mm in diameter were made in the calvaria of the rats. The animals were divided into two groups: Bone Defect Control Group (GCDO) and Experimental Immediate Vibration Group (GEVI). GEVI animals were submitted to 60 Hz vibrating waves and 0.3 g vertical acceleration; they were applied three times/week, for twenty minutes. Fifteen days after the surgery, the animals were euthanized to measure the extent of the defect. Considering that these defects had the same initial diameter, a reduction in their final linear extension was admitted as an indirect indicator of osteoid deposition. Results: neoformation of an osteoid matrix, restricted to bone borders, was observed in both groups. The mean linear extension, in millimeters, of the GEVI bone defect was 5.83 (SD = 0.79) and in the GCDO, it was 6.62 (SD = 0.63). There was no statistically significant difference between the means (U = 8.00, z = -1.604, p = 0.132). Conclusion: an osteogenic response was evidenced from the use of vibratory therapy, however in a statistically non-significant way. Thus, the present study demonstrated that the use of vibrating waves did not favor a statistically significant bone repair, during the studied period and vibration regime

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.

Dental Pulpal Tissue Regeneration, Pulpal Vitality Testing, and Healing of Apical Lesions Following Stem Cell Transplant: A Systematic Review and Meta-Analysis

Abstract Objective: To analyze data obtained from animal and human studies using stem cells. Material and Methods: Medical Literature Analysis and Retrieval System Online (MEDLINE), PubMed, Cochrane Library, Embase, Information Sciences Institute (ISI), as well as Google Scholar were utilized and searched as available electronic databases to perform a systematic literature review of articles published between 2010 and 2019. The Endnote X9 for Windows was also employed to manage electronic titles and abstracts of the selected studies. Searches were conducted using keywords of "pulpal OR pulpal tissue OR pulpal vitality", "regeneration", "apical healing", "stem cells OR progenitor cells", and "mediated pulpal tissue". Consequently, 189 titles and abstracts endowed with potential relevance were discovered based on searches into manual and electronic sources. Ultimately, a total of six articles met the inclusion criteria in the present systematic review and meta-analysis. Results: Out of the six articles identified and selected, five studies were categorized as animal experiments and one article was nominated as a human clinical trial. The greatest bias risks were accordingly observed in the majority of animal examinations, but articles related to humans revealed decreased risks of bias, while the human clinical trial showed some concerns. Conclusion: Promising parameters testing functional pulp regeneration could be represented through stem cell transplants.

Efeito da Justicia acuminatissima, Sara Tudo do Amazonas, na injúria renal aguda isquêmica: estudo experimental / Efecto de la planta Justicia acuminatissima, "Sana Todo del Amazonas", en la injuria renal aguda isquémica: estudio experimental / Effects of Justicia acuminatissima, or Amazonian Sara Tudo, on ischemic acute kidney injury: an experimental study

RESUMO Objetivo Avaliar o efeito da Justicia acuminatissima , Sara Tudo do Amazonas, na função renal, na hemodinâmica renal, no perfil oxidativo e na histologia renal em ratos com injúria renal aguda isquêmica. Método Ensaio pré-clínico com ratos Wistar, adultos, machos (250-350 g), distribuídos nos grupos Sham, Isquemia e Isquemia + Sara Tudo. Foram avaliados os parâmetros hemodinâmicos, a função renal, o estresse oxidativo e a histologia renal. Resultados O pré-tratamento com o Sara Tudo atenuou a lesão funcional, o que foi evidenciado pelo aumento no clearance de creatinina, redução dos marcadores oxidativos e elevação de tióis, pela melhora significativa do fluxo sanguíneo renal, diminuição da resistência vascular renal e redução da lesão tubulointersticial no tecido renal. Conclusão A renoproteção da Justicia acuminatissima , Sara Tudo, na injúria renal aguda isquêmica, caracterizou-se por melhora significativa da função renal, reduzindo a lesão oxidativa, com impacto positivo na histologia renal.

RESUMEN Objetivo Evaluar el efecto de la planta Justicia acuminatissima , "Sana Todo del Amazonas", en la función renal, la hemodinámica renal, el perfil oxidativo y la histología renal en ratones con injuria renal aguda isquémica. Método Ensayo pre clínico con ratones Wistar, adultos, machos (250-350 g), distribuidos en los grupos Sham, Isquemia e Isquemia + Sana Todo. Fueron evaluados los parámetros hemodinámicos, la función renal, el estrés oxidativo y la histología renal. Resultados El pre tratamiento con el Sana Todo atenuó la lesión funcional, lo que fue evidenciado por el aumento en el aclaramiento de creatinina, reducción de los marcadores oxidativos y elevación de tioles, por la mejora significativa del flujo sanguíneo renal, disminución de la resistencia vascular renal y reducción de la lesión tubulointersticial en el tejido renal. Conclusión La renoprotección de la Justicia acuminatissima , "Sana Todo del Amazonas", en la injuria renal aguda isquémica se caracterizó por mejora significativa de la función renal, reduciendo la lesión oxidativa, con impacto positivo en la histología renal.

ABSTRACT Objective To evaluate the effects of Justicia acuminatissima , or Amazonian Sara Tudo , on renal hemodynamics, oxidative profile, and renal histology in rats with ischemic acute kidney injury. Method Preclinical assay with adult male Wistar rats, weighing from 250 g to 350 g, distributed into Sham, ischemia, and ischemia + Sara Tudo groups. Hemodynamic parameters, renal function, oxidative stress, and renal histology were evaluated. Results Pretreatment with Sara Tudo reduced the functional injury, which was shown by the increase in creatinine clearance and thiols; reduction of oxidative markers, renal vascular resistance, and tubulointerstitial injury in the renal tissue; and the significant improvement in renal blood flow. Conclusion The renoprotection provided by Justicia acuminatissima , or Sara Tudo , in cases of ischemic acute kidney injury was characterized by a marked improvement in renal function, reducing the oxidative injury, and impacting on renal histology positively.

Analysis of Estrogen Effects on Eruption of Rat Molars / Análisis de los Efectos del Estrógeno en la Erupción de Molares de Rata

ABSTRACT: Tooth eruption requires resorption of the alveolar bone interposed between the tooth germ and the oral mucosa (coronal bone). The cells responsible for bone resorption are the osteoclasts and their activity can be reduced or inactivated by estrogen hormone. We aimed to investigate the effects of estrogen on the process of tooth eruption in rats. Thirty-three Wistar rats, aged two-to-17-days, were divided into control, sham and estrogen-treated groups. After daily injections with estrogen, the animals were euthanized and the jaws removed and processed for histological analysis. We performed clinical examination, morphological analysis, quantification of the number of osteoclasts on the surface of the coronal bone and immunohistochemical analysis of estrogen receptor type alpha (ERα). Estrogen therapy was effective, which could be confirmed by the higher estrogen plasma levels on treated animals. However, it had no effect on tooth development or tooth eruption. Progressive bone resorption was observed and the number of osteoclasts on coronal bone was not affected on hormoneinjected animals, allowing tooth to erupt at the same time observed in untreated animals. Immunohistochemistry for ERα confirmed the presence of this type of receptor in osteoclasts, osteoblasts and osteocytes. Taken together, our results showed that estrogen stimulation was not sufficient to decrease the number of osteoclasts on the coronal bone, supporting the idea that, although estrogen may have a protective activity on bone resorption, this may not apply to the alveolar bone that is meant to be resorbed during eruptive process.

RESUMEN: La erupción dental requiere la resorción del hueso alveolar interpuesto entre el germen dental y la mucosa oral (hueso coronal). Las células responsables de la resorción ósea son los osteoclastos y su actividad puede reducirse o inactivarse por la hormona del estrógeno. Objetivos: apuntamos a investigar los efectos del estrógeno en el proceso de la erupción dental en ratas. Treinta y tres ratas Wistar, de dos a 17 días de edad, se dividieron en grupos de control, Sham y se trataron con estrógenos. Los animales fueron eutanizados después del tratamento con estrógeno y se procesaron las mandíbulas para el análisis histológico. Se realizó el examen clínico, el análisis morfológico, la cuantificación del número de osteoclastos en la superficie del hueso coronal y el análisis inmunohistoquímico del tipo de receptor de estrógeno alfa (ERα). La terapia de estrógeno fue eficaz, lo que podría ser confirmado por los niveles plasmáticos más altos de estrógeno en los animales tratados. Sin embargo, no se observó ningún efecto sobre el desarrollo de los dientes o la erupción dental. Se observó una resorción ósea progresiva y el número de osteoclastos en el hueso coronal no se vio afectado en los animales inyectados con hormonas, permitiendo que el diente erupcionó durante el mismo período de tiempo observado en animales no tratados. La inmunohistoquímica para el ERα confirmó la presencia de este tipo de receptor en los osteoclastos, osteoblastos y osteocitos. Nuestros resultados mostraron que la estimulación del estrógeno no fue suficiente para reducir el número de osteoclastos en el hueso coronal confirmando que, si bien el estrógeno puede tener una actividad protectora en la resorción ósea, esto puede no se aplica al hueso alveolar que está destinado a ser rerecurrido durante el proceso eruptivo.

Functional Features of Hemocoagulation in rats with Experimentally formed Arterial Hypertension in Conditions of Increased Motor Activity

In rats with experimentally formed arterial hypertension, lipid perxidation in the plasma, amplification of blood chotting mechanisms with a decrease in anticoagulation and fibrinolysis was noted. Regular forced jogging provided the experimental rats with a positive dynamic of all the indicators considered. Thus, with increased muscular activity, the level of acyl hydro-peroxides of plasma decreased in rats with arterial hypertension formed due to the enhancement of its antioxidant activity. In addition, with the increase in muscle activity in experimental rats, normalization of clotting factor activity, indices of general coagulation tests, antithrobin III activity and protein C was achieved. This was accompanied by a normalization of the level of plasminogen, a2-antiplasmin and spontaneous euglobulin lysis time. In rats with formed arterial hipertension with stgandard physical activity, the initial violations of the measured parameters were completely preserved (AU)