Using in vivo animal models for studying SARS-CoV-2.

INTRODUCTION: The search for an animal model capable of reproducing the physiopathology of the COVID-19, and also suitable for evaluating the efficacy and safety of new drugs has become a challenge for many researchers. AREAS COVERED: This work reviews the current animal models for in vivo tests with SARS-CoV-2 as well as the challenges involved in the safety and efficacy trials. EXPERT OPINION: Studies have reported the use of nonhuman primates, ferrets, mice, Syrian hamsters, lagomorphs, mink, and zebrafish in experiments that aimed to understand the course of COVID-19 or test vaccines and other drugs. In contrast, the assays with animal hyperimmune sera have only been used in in vitro assays. Finding an animal that faithfully reproduces all the characteristics of the disease in humans is difficult. Some models may be more complex to work with, such as monkeys, or require genetic manipulation so that they can express the human ACE2 receptor, as in the case of mice. Although some models are more promising, possibly the use of more than one animal model represents the best scenario. Therefore, further studies are needed to establish an ideal animal model to help in the development of other treatment strategies besides vaccines.

Tannic Acid Solution: A Better Fixative Solution Than Formalin for Elastin and Collagen-Toxic and Morphological Assessment.

Formaldehyde is commonly used worldwide, even though it is classified as carcinogenic to humans by the International Agency for Research on Cancer. This has motivated intensive investigations of formaldehyde substitutes, and recently, some alternative solutions were found, which can potentially replace it. Previous research showed that tannic acid (TA) in glutaraldehyde solution has the ability to stabilize elastin and collagen. This provided a basis for the development of a new alcoholic fixative solution, particularly aimed at extracellular matrix components, with TA as a main component. Heart, brain, and intestinal samples were fixed by immersion in 10% regular formalin solution (RFS), 70% ethanol solution (ES), and tannic acid ethanolic solution (TAES). Next, tissue fragments were prepared for routine histology procedures. The toxicity of TA was analyzed using in silico tests for mutagenicity, as well as for cutaneous and respiratory toxicity. Analyses of photomicrographs demonstrated that all fixative solutions have the ability to preserve the fragments. The quantitative analyses showed that capability of TAES to preserve and stabilize elastin and collagen is superior to that of RFS and ES. We demonstrated that TA is not mutagenic, and it is less toxic for skin and respiratory tract. We therefore conclude that TAES can potentially represent a powerful and feasible alternative solution for fixing extracellular matrix of microscopic examination samples. Anat Rec, 301:1544-1550, 2018. © 2018 Wiley Periodicals, Inc.

Clinical discussion of the arteria lusoria: a case report / Discussão clínica a respeito da artéria lusória: um relato de caso

Abstract The right subclavian artery may originate from the left portion of the aortic arch. This aberrant vessel is known as the arteria lusoria. Its course to its usual site runs behind the esophagus, which may cause a disease known as dysphagia lusoria, responsible for symptoms of discomfort. This artery is often associated with other anomalies, such as the non-recurrent laryngeal nerve and the bicarotid trunk, and with diseases such as aneurysms, congenital heart defects, and even genetic syndromes. During routine dissection of a male cadaver fixed in 10% formalin solution, an arteria lusoria was found. This article reports the variation and discusses its embryological, clinical and surgical aspects.

Resumo Em alguns casos, a artéria subclávia direita pode se originar da porção esquerda do arco aórtico. Esse vaso aberrante é conhecido como artéria lusória. Para chegar em sua região, essa artéria passa posteriormente ao esôfago, e pode ser, portanto, causa de uma doença conhecida como disfagia lusória, desencadeando sintomas desconfortantes. A artéria lusória está frequentemente associada com outras anomalias, como o nervo laríngeo não recorrente e o tronco bicarotídeo, assim como pode estar em associação com aneurismas, defeitos cardíacos congênitos e até síndromes genéticas. Durante dissecação cadavérica de rotina, foi observada a presença dessa artéria em um cadáver do sexo masculino fixado em uma solução de formalina a 10%. O objetivo deste trabalho é relatar a variação conhecida como artéria lusória e trazer destaque para seus aspectos embriológicos, clínicos e cirúrgicos.

Clinical discussion of the arteria lusoria: a case report.

The right subclavian artery may originate from the left portion of the aortic arch. This aberrant vessel is known as the arteria lusoria. Its course to its usual site runs behind the esophagus, which may cause a disease known as dysphagia lusoria, responsible for symptoms of discomfort. This artery is often associated with other anomalies, such as the non-recurrent laryngeal nerve and the bicarotid trunk, and with diseases such as aneurysms, congenital heart defects, and even genetic syndromes. During routine dissection of a male cadaver fixed in 10% formalin solution, an arteria lusoria was found. This article reports the variation and discusses its embryological, clinical and surgical aspects.

Em alguns casos, a artéria subclávia direita pode se originar da porção esquerda do arco aórtico. Esse vaso aberrante é conhecido como artéria lusória. Para chegar em sua região, essa artéria passa posteriormente ao esôfago, e pode ser, portanto, causa de uma doença conhecida como disfagia lusória, desencadeando sintomas desconfortantes. A artéria lusória está frequentemente associada com outras anomalias, como o nervo laríngeo não recorrente e o tronco bicarotídeo, assim como pode estar em associação com aneurismas, defeitos cardíacos congênitos e até síndromes genéticas. Durante dissecação cadavérica de rotina, foi observada a presença dessa artéria em um cadáver do sexo masculino fixado em uma solução de formalina a 10%. O objetivo deste trabalho é relatar a variação conhecida como artéria lusória e trazer destaque para seus aspectos embriológicos, clínicos e cirúrgicos.

De Garengeot's hernia in an 82-year-old man: a case report and clinical significance.

The presence of the appendix within a femoral hernia (FH) sac is known as Garengeot's hernia (GH). We report on current study a rare case of an elderly man with a combined inguinal and Garengeot's hernia and discuss the clinical aspects. An 82-year-old man clinically stable, presented history of pain at the right inguinal region for over a week, without vomit, nausea, fever or any alteration of intestinal or urinary eliminations. Clinical examination revealed a FH and the ultrasonography confirmed the hernia sac. During the surgery, the appendix was recognized within the sac, and then, the patient underwent appendectomy and hernia repair. In conclusion, the presence of the vermiform appendix in a FH sac is rare, thus, requiring knowledge of the surgeon regarding this clinical entity. Prompt diagnosis and appropriate surgical treatment is the key to avoid complications.

Pharyngoesophageal Obstruction on the Killian-Laimer Triangle by Zenker's Diverticulum: Case Report and Clinical Significance.

Zenker's diverticulum is a form of esophageal and pharyngeal obstruction located at the Killian-Laimer triangle. It is relatively common in elderly man (seventh or eighth decade of life), and its pathophysiology is not completely understood, albeit theories regarding dysfunction of the upper esophageal sphincter were reported. The main symptoms are dysphagia and odynophagia, but it can complicate to aspiration and perforation of the pharyngeal pouch; also, it can be asymptomatic. Diagnosis is provided through a barium esophagogram. Treatment can be performed through endoscopic surgery, diverticulopexy and myotomy of the cricopharyngeus muscle, although there is no consensus among surgeons regarding the treatment of choice. We report a case of pharyngeal obstruction due to Zenker's diverticulum which caused massive weight loss in a 76-year-old male.

Maternal malnutrition during lactation in Wistar rats: effects on elastic fibers of the extracellular matrix in the trachea of offspring.

OBJECTIVE: To investigate the effects of maternal protein malnutrition during lactation on the elastic fibers in the tracheas of Wistar rat pups. METHODS: At delivery, 12 male pups of two Wistar rat dams were equally divided into two groups: control, in which the dam received water and standard rat chow ad libitum during lactation; and protein-restricted (PR), in which the dam received water ad libitum and an isoenergetic PR diet (8% protein). At 21 days of age, the pups were killed and their tracheas were excised. The elastic fibers were stained with Weigert's resorcin-fuchsin (after oxidation) and evaluated under light microscopy. Morphometric determinations were performed by stereology, with the point-counting method, and expressed as volumetric densities. RESULTS: Elastic fibers, most having a longitudinal distribution, were identified beneath the tracheal mucosa. In addition, well-defined circular layers of elastic fibers were found around the inner and outer surfaces of the cartilaginous ring. There were no differences between the groups regarding the organization and distribution of the elastic fibers. The volumetric density of the elastic fibers of the pups in the control and PR groups was 2.46 ± 0.99% and 3.25 ± 1.13%, respectively (p < 0.01). CONCLUSIONS: The volumetric density of elastic fibers appears to be greater in rat pups breastfed by dams receiving a PR diet than in those breastfed by dams receiving a normal diet.

Desnutrição materna durante a lactação em ratos Wistar: efeitos sobre as fibras elásticas da matriz extracelular na traqueia dos filhotes / Maternal malnutrition during lactation in Wistar rats: effects on elastic fibers of the extracellular matrix in the trachea of offspring

OBJETIVO: Investigar os efeitos da desnutrição proteica materna durante a lactação sobre as fibras elásticas da traqueia de filhotes de ratos Wistar. MÉTODOS: Ao nascimento, 12 filhotes machos de duas ratas Wistar foram igualmente divididos em dois grupos: grupo controle, cuja mãe recebeu água e dieta padrão de laboratório ad libitum durante a lactação, e grupo restrição proteica (RP), cuja mãe recebeu água ad libitum e dieta isoenergética com RP (8% de proteína). Aos 21 dias de vida, os filhotes foram sacrificados, e suas traqueias foram ressecadas. As fibras elásticas foram coradas pelo método de resorcina-fucsina de Weigert (precedido de oxidação) e avaliadas sob microscopia óptica. As determinações morfométricas foram feitas por estereologia, utilizando o método de contagem de pontos, e expressas em densidade volumétrica. RESULTADOS: As fibras elásticas foram identificadas abaixo da mucosa traqueal, sendo a maioria em distribuição longitudinal. Além disso, camadas circulares bem definidas de fibras elásticas envolviam as superfícies interna e externa do anel cartilaginoso. Não houve diferenças entre os grupos quanto à organização e distribuição das fibras elásticas. A densidade volumétrica das fibras elásticas dos filhotes nos grupos controle e RP foi de, respectivamente, 2,46 ± 0,99% e 3,25 ± 1,13% (p < 0,01). CONCLUSÕES: Nossos resultados sugerem que a densidade volumétrica de fibras elásticas é maior em filhotes de ratos alimentados por fêmeas submetidas a dieta com RP do que naqueles de mães recebendo dieta normal.

OBJECTIVE: To investigate the effects of maternal protein malnutrition during lactation on the elastic fibers in the tracheas of Wistar rat pups. METHODS: At delivery, 12 male pups of two Wistar rat dams were equally divided into two groups: control, in which the dam received water and standard rat chow ad libitum during lactation; and protein-restricted (PR), in which the dam received water ad libitum and an isoenergetic PR diet (8% protein). At 21 days of age, the pups were killed and their tracheas were excised. The elastic fibers were stained with Weigert's resorcin-fuchsin (after oxidation) and evaluated under light microscopy. Morphometric determinations were performed by stereology, with the point-counting method, and expressed as volumetric densities. RESULTS: Elastic fibers, most having a longitudinal distribution, were identified beneath the tracheal mucosa. In addition, well-defined circular layers of elastic fibers were found around the inner and outer surfaces of the cartilaginous ring. There were no differences between the groups regarding the organization and distribution of the elastic fibers. The volumetric density of the elastic fibers of the pups in the control and PR groups was 2.46 ± 0.99% and 3.25 ± 1.13%, respectively (p < 0.01). CONCLUSIONS: The volumetric density of elastic fibers appears to be greater in rat pups breastfed by dams receiving a PR diet than in those breastfed by dams receiving a normal diet.

Structure and concentration of elastic system fibers in the trachea of the rat / Estructura y concentración del sistema de fibras elásticas en la tráquea de la rata

The extracellular matrix is a key element in the tracheal morphology and physiology, both in normal and pathological states, yet little is known regarding its composition and organization. Herein we carried out a detailed study of the morphological organization and volumetric density of elastic system fibers in the trachea of Wistar rats. Six Wistar rats at the age of 45 days were used. The trachea was excised following anesthesia with Thiopental and perfusion through the left ventricle with buffered saline followed by formalin solution. Samples were fixed in formaline, embedded in paraffin, and histologically processed. The elastic system fibers were evaluated at light microscopy by using Weigert's resorcin-fuchsin technique after oxidation with oxone. Morphometric studies were performed by the point-counting method. Quantities were expressed ( percent, mean) as volumetric densities and were determined on 25 random fields for each animal. Elastic system fibers were identified beneath the tracheal mucosa showing mainly a longitudinal profile. There were also some bundles of oblique fibers in this region, forming an irregular network of elastic tissue. Close to the inner surface of the cartilaginous ring, a well defined circular layer is present. The volumetric density of the elastic system fibers in the trachea of Wistar rats is 2.46 +/- 0.99 percent. The data in the present study provides original information regarding the elastic system fibers of the rat trachea, which might be used as a model for studying human major airway morphology. The results reported herein provide the basis for continuous investigations on tracheal extracellular matrix by stereology.

La matriz extracelular es un elemento clave en la morfología y fisiología de la tráquea, tanto en estados normales como patológicos. Sin embargo, poco se sabe acerca de su composición y organización. Se llevó a cabo un estudio detallado de la organización morfológica y la densidad volumétrica de fibras del sistema elástico en la tráquea de ratas Wistar. Se utilizaron seis ratas Wistar edad de 45 días de vida. La tráquea fue extraída después de anestesia con tiopental y la perfusión a través del ventrículo izquierdo con solución salina tamponada seguido de solución de formalina. Las muestras fueron fijadas en formalina e incluidas en parafina y se procesaron histológicamente. Las fibras del sistema elástico fueron evaluadas en microscopía de luz utilizando la técnica de Weigert, resorcina-fucsina después de la oxidación con Oxone. Los estudios morfométricos fueron realizados por el método de conteo de puntos. Las cantidades se expresaron ( por ciento, media) como la densidad volumétrica y se determinaron en 25 campos al azar para cada animal. Las fibras elásticas del sistema fueron identificadas por bajo la mucosa traqueal que muestra principalmente un perfil longitudinal. También se encontraron algunos haces de fibras oblicuas en esta región, formando una red irregular de tejido elástico. Cerca de la superficie interna del anillo cartilaginoso, una capa bien definida circular está presente. La densidad volumétrica de las fibras del sistema elástico en la tráquea de ratas Wistar fue de 2,46 +/- 0,99 por ciento. Los datos del presente estudio proporcionan información inicial sobre las fibras del sistema elástico de la tráquea de rata, que puede ser utilizado como un modelo para el estudio de la morfología de las vías respiratorias humanas. Los resultados reportados en este documento constituyen la base de investigaciones continuas en la tráquea de la matriz extracelular por estereología.