Extraction of DNA from plastinated tissues.

INTRODUCTION: Plastination allows anatomical samples to be preserved in excellent condition for an indefinite period, free of formalin, and in a format that allows biosafe manipulation by students, academics, and researchers. As with other tissue preservation techniques, it is important to establish the level of conservation of deoxyribonucleic acid (DNA) for use in future applications. The object of the present work was to extract and evaluate DNA from plastinated tissues. METHODS: We used samples of liver from Canis lupus familiaris and skeletal muscle from Rattus norvegicus, Sprague-Dawley strain, extracted from specimens plastinated with silicone at room temperature. The tissue samples were deplastinated by incubation in 5% sodium methoxide dissolved in methanol for 24 or 48 h. The samples were divided into two equal parts and DNA was extracted using two different protocols. After extraction, the DNA was quantified by fluorometry and its integrity was assessed by electrophoresis in a 1% agarose gel. RESULTS AND DISCUSSION: A high yield of DNA was obtained from the deplastinated samples and the DNA was intact. Plastinated tissues have proven to be stable and easily managed. They can also be used for examination under light and electron microscopes. The DNA extraction technique used here allowed us to obtain intact DNA from samples plastinated with silicone at room temperature, without previous fixing. This technique may allow tissue specimens to be preserved for retrospective studies of archived samples of normal and pathological anatomy in the fields of basic, clinical, forensic, and epidemiological sciences. CONCLUSIONS: The extracted DNA was intact and suitable for use in subsequent applications. Obtaining whole DNA from plastinated samples using tissue preservation protocols that preserve the tissue for use in subsequent applications, like real-time PCR, opens up many possibilities, with applications in the basic and clinical sciences, epidemiology, and forensic science.

Development of an ultrathin sheet plastination technique in rat humeral joints with osteoarthritis induced by monosodium iodoacetate for neovascularization study.

Injection with monosodium iodoacetate (MIA) is widely used to produce osteoarthritis (OA). Ultrathin sheet plastination has been used to study the morphology of structures, with strong application in anatomical education and research. Our aim was to carry out, for the first time, ultrathin sheet plastination of rat humeral joints to observe the neovascularization provoked by OA. We injected 0.1 mL of MIA into the left humeral joints of ten Sprague-Dawley rats. The right shoulders of the same rats were used as control. Sixteen weeks after the injection, the animals were euthanized and were given an immediate red epoxy resin injection through the thoracic aorta. The samples were fixed in 10% formalin, prior to the plastination process, without decalcification. Samples were dehydrated with acetone (100%) at - 25 °C, for 10 days. Later, for degreasing, samples were immersed in methylene chloride at room temperature during 1 week. Forced impregnation was performed inside a stove within a vacuum chamber. The plastinated blocks obtained were cut with a slow velocity diamond blade saw. Slices were placed in curing chambers to achieve curing and final tissue transparentation. 230 µm thickness slices were obtained. The slices were analyzed under magnifying glass and microscope, achieving visualization of OA neovascularization. The cartilage affected by OA loses its ability to remain avascular, and blood vessels invade it from the subchondral bone to the calcified and uncalcified cartilage. Ultra-thin sheet plastination is useful to observe articular cartilage neovascularization, caused by OA induced with MIA in humeral rat joint.

P40 polyester sheet plastination technique for brain and body slices: The vertical and horizontal flat chamber methods.

The P40 technique produces high-quality brain and body slices and is the user-friendliest of the polyester techniques. The P40 polyester technique follows the same classical steps for plastination. That is, preparation of the specimen, fixation (optional), dehydration by freeze substitution, forced impregnation and curing. Two methods used to prepare two different types of specimens, that is, brain slices and body slices are described. Each method has its own characteristics depending on the specimen type used. Brain slices were used to illustrate the vertical small chamber method while the body slices were used to illustrate the horizontal large chamber method. The brain slices obtained using P40 are of very good quality presenting good contrast between grey and white matter. The body slices are also of very good quality. The physical appearance of these slices makes them an exceptional instrument for diagnostic imaging and anatomical correlation. Body slices prepared with P40 retain the natural colour of the tissue and preserve the anatomical relationships.

E12 sheet plastination: Techniques and applications.

Plastination is an anatomical technique that consists of replacing the liquids and fat of specimens by reactive polymers through forced impregnation in a vacuum. These are then polymerized to achieve the final result. E12 sheet plastination involves epoxy resin impregnation of thin (2-4 mm) and ultra-thin (<2 mm) tissue sheets, producing dry, transparent, odorless, non-toxic and long-lasting sheets. E12 sheet plastination techniques were reviewed using MEDLINE, EMBASE and SciELO databases, and manual searches. After searching, 616 records were found using the online and manual searches (MEDLINE, n: 207; EMBASE, n: 346; SciELO, n: 44; Manual search: 23). Finally, 96 records were included in this review (after duplicates and articles unrelated to the subject were excluded). The aim of this work was to review the E12 sheet plastination technique, searching for articles concerning views of it, identifying the different variants implemented by researchers since its creation by Gunther von Hagens, and to identify its applications from teaching and research in anatomy to morphological sciences. Clin. Anat. 31:742-756, 2018. © 2017 Wiley Periodicals, Inc.

Chagas disease: a proposal for testing policy for solid-organ transplant in the United States.

Chagas disease is caused by the parasite Trypanosoma cruzi. The disease is difficult to detect because of the asymptomatic pathogenesis after infection. Chagas disease is endemic throughout much of Mexico, Central America, and South America, but human migration patterns are bringing the pathogen to the United States. The disease currently affects 16 to 18 million people with approximately 50 000 deaths annually in these countries. In the United States, national screening of the blood supply was instituted in early 2007, and more than 1000 donors with T cruzi infection have been identified within the past 3 years of testing. It was observed that out of the 58 organ procurement organizations in the United States, only 4 required mandatory testing of every donor for Chagas disease. It was estimated that as of 2009, approximately 409 000 residents are living with Chagas disease, and in a 22-year span, approximately 300 patients may have contracted Chagas disease through transplant. Proposed solutions to the current testing method include automatic testing based on the medical social history questionnaire, testing of all recipients for Chagas disease, testing all persons of Latin descent, or testing of all organ donors.

Impregnation of biological specimes with resins and elastowers: plastination with biodur S 10 resin

Várias säo as desvantagens do uso de formaldeídeo, álcool glicerina e outros líquidos para a conservaçäo de peças anatômicas. O método da plastinaçäo se mostra superior aos métodos tradicionais de conservaçäo em líquidos e consiste de quatro etapas fundamentais: fixaçäo, desidrataçäo impregnaçäo e endurecimento. No presente foi utilizada a resina Biodur S10 adicionada do respectivo catalisador. As peças plastinadas resultantes apresentaram grande flexibilidade, aparência quase natural e de fácil manuseio