Comparación de resultados entre soluciones con y sin formol como medios de fijación y conservación en corazones de Gallus gallus domesticus / Comparison of results between solutions with and without formaldehyde as means of fixation and conservation in Gallus gallus domesticus hearts

RESUMEN: La solución de formol es utilizada en las Escuelas de medicina como medio de fijación y conservación de cadáveres para el estudio de la Anatomía, a la que están expuestos estudiantes, técnicos y personal docente; es alergénica e irritante a las mucosas, y reconocida carcinogénica en humanos por International Agency for Research on Cancer (2006). El objetivo del presente estudio fue comparar resultados cuantitativos y cualitativos entre corazones de Gallus gallus domesticus, luego de aplicarles soluciones con y sin formol. Se formaron dos grupos al azar, a uno se le aplicó solución de formol al 10 %, y al otro solución libre de formol. Se realizaron medidas antropométricas, organolépticas, y de fotografía (Pretest, durante y Postest). Se elaboró base datos en Microsoft Excel (2019), y su procesamiento en SPSS Statistics 2017 Versión 25. Para variables cuantitativas se aplicó la prueba de Shapiro-Wilk, y t-Student pareada. Para variables cualitativas el test Alfa de Cronbach, Chi cuadrado (X2) y los correspondientes coeficientes de asociación (D de Somers y Tau b de Kendal). Los resultados obtenidos de las variables peso, largo, y altura presentaron diferencia estadística significativa (p-valor <0,05), siendo diferente para el ancho y grosor de la pared del ventrículo izquierdo. Las variables color y consistencia presentaron diferencias significativa (p-valor <0,05). El olor irritante a las mucosas estuvo presente durante todo el estudio con la solución con formol. A la inspección, ninguno de los dos grupos presento colonización - descomposición. Se concluye que, los órganos en experimentación que se les aplicó solución libre de formol, presentaron mejores resultados con respecto a los que se les aplico formol al 10 %.

SUMMARY: The formaldehyde solution is used in medical schools as a means of fixing and preserving corpses for the study of Anatomy, to which students, technicians and teaching personnel are exposed; it is allergenic and irritant to the mucosa, and recognized as a human carcinogen by the International Agency for Research on Cancer (2006). The objective of the present study was to compare quantitative and qualitative results between Gallus gallus domesticus hearts, after applying solutions with and without formaldehyde. Two groups were formed at random, to one a 10 % formaldehyde solution was applied, and to the other formaldehyde- free solution. Anthropometric, organoleptic, and photographic measurements were carried out (Pretest, during and Posttest). A database was prepared in Microsoft Excel (2019), and its processing in SPSS Statistics 2017 Version 25. For quantitative variables, the Shapiro-Wilk test and t-Student paired were applied. For qualitative variables the Cronbach's Alpha test, Chi square (X2) and the corresponding association coefficients (Somers D and Kendal's Tau b). The results obtained from the variables weight, length, and height presented a statistically significant difference (p-value <0.05), being different for the width and thickness of the left ventricular wall. The variables color and consistency showed significant differences (p-value <0.05). The irritating smell to the mucous membranes was present throughout the study with the formaldehyde solution. Upon inspection, neither group showed colonization - decomposition. It is concluded that the organs in experimentation that were applied formaldehyde-free solution presented better results compared to those that were applied 10 % formaldehyde.

Difference between actual vs. pathology prostate weight in TURP and radical robotic-assisted prostatectomy specimen

Introduction To investigate and highlight the effect of formaldehyde induced weight reduction in transurethral resection of prostate (TURP) and radical robotically-assisted prostatectomy (RALP) specimen as a result of standard chemical fixation. Materials and Methods 51 patients were recruited from January 2013 to June 2013 who either underwent a TURP (n=26) or RALP (n=25). Data was collected prospectively by the operating surgeon who measured the native, unfixed histology specimen directly after operation. The specimens were fixed in 10% Formaldehyde Solution BP and sent to the pathology laboratory where after sufficient fixation period was re-weighed. Results Overall mean age 64.78 years, TURP mean age 68.31 years RALP mean age 61.12years. We found that the overall prostatic specimen (n=51) weight loss after fixation was a mean of 11.20% (3.78 grams) (p≤0.0001). Subgroup analysis of the native TURP chips mean weight was 16.15 grams and formalin treated mean weight was 14.00 grams (p≤0.0001). Therefore, TURP chips had a mean of 13.32 % (2.15 grams) weight loss during chemical fixation. RALP subgroup unfixed specimen mean weight was 52.08 grams and formalin treated mean weight was 42.60 grams (p≤0.0001), a 19.32 % (9.48grams) mean weight reduction. Conclusion It has not been known that prostatic chips and whole human radical prostatectomy specimen undergo a significant weight reduction. The practical significance of the accurate prostate weight in patient management may be limited, however, it is agreed that this should be recorded correctly, as data is potential interest for research purposes and vital for precise documentation. .

Estudo in vivo do comportamento de bioprótese liofilizada: seguimento de 3 meses em carneiros jovens / In vivo study of lyophilized bioprostheses: 3 month follow-up in young sheep

OBJETIVO: Para melhorar as propriedades mecânicas e imunogênicas, o glutaraldeído é utilizado no tratamento do pericárdio bovino que é utilizado em biopróteses. A liofilização do pericárdio bovino tratado com glutaraldeído diminui os radicais aldeído, com provável redução do potencial para calcificação. O objetivo deste estudo é avaliar os efeitos da liofilização em biopróteses valvares de pericárdio bovino como mecanismo protetor na diminuição da disfunção estrutural valvar. MÉTODOS: Foi realizado o implante de biopróteses de pericárdio bovino tratado com glutaraldeído, liofilizadas ou não, em carneiros de 6 meses de idade, sendo os animais eutanasiados com 3 meses de seguimento. As biopróteses foram implantadas em posição pulmonar, com auxílio de circulação extracorpórea. Um grupo controle e outro grupo liofilizado foram avaliados quanto ao gradiente ventrículo direito/artéria pulmonar (VD/AP) no implante e explante; análise quantitativa de cálcio; inflamação; trombose e pannus. O nível de significância estabelecido foi de 5%. RESULTADOS: O gradiente médio VD/AP, no grupo controle, no implante, foi 2,04 ± 1,56 mmHg e, no grupo de liofilização, foi 6,61 ± 4,03 mmHg. No explante, esse gradiente aumentou para 7,71 ± 3,92 mmHg e 8,24 ± 6,2 mmHg, respectivamente, nos grupos controle e liofilização. O teor de cálcio médio, após 3 meses, nas biopróteses do grupo controle foi 21,6 ± 39,12 µg Ca+2/mg de peso seco, em comparação com um teor médio de 41,19 ± 46,85 µg Ca+2/mg de peso seco no grupo liofilizado (P = 0,662). CONCLUSÃO: A liofilização de próteses valvares com pericárdio bovino tratado com glutaraldeído não demonstrou diminuição da calcificação neste experimento.

OBJECTIVE: Glutaraldehyde is currently used in bovine pericardium bioprosthesis to improve mechanical and immunogenic properties. Lyophilization is a process that may decrease aldehyde residues in the glutaraldehyde treated pericardium decreasing cytotoxicity and enhancing resistance to calcification. The aim of this study is to evaluate bioprosthetic heart valves calcification in adolescent sheep and to study the potential of lyophilization as a mechanism to protect calcification. METHODS: Two groups were evaluated: a control group in which a bovine pericardium prosthetic valve was implanted in pulmonary position and a lyophilized group in which the bovine pericardium prosthetic valve was lyophilized and further implanted. Sixteen sheeps 6 months old were submitted to the operation procedure. After 3 months the sheeps were euthanized under full anesthesia. RESULTS: Six animals of the control group reached 95.16 ± 3.55 days and six animals in the lyophilized group reached 91.66 ± 0.81 days of postoperative evolution. Two animals had endocarditis. Right ventricle/pulmonary artery (RV/PA) mean gradient, in the control group, at the implantation was 2.04 ± 1.56 mmHg, in the lyophilization group, the RV/PA mean gradient, at the implantation was 6.61 ± 4.03 mmHg. At the explantation it increased to 7.71 ± 3.92 mmHg and 8.24 ± 6.25 mmHg, respectively, in control and lyophilization group. The average calcium content, after 3 months, in the control group was 21.6 ± 39.12 µg Ca+2/mg dry weight, compared with an average content of 41.19 ± 46.85 µg Ca+2/mg dry weight in the lyophilization group (P=0.662). CONCLUSION: Freeze drying of the bovine pericardium prosthesis in the pulmonary position could not demonstrate calcification mitigation over a 3 month period although decreased inflammatory infiltration over the tissue.

Efeito da descelularização com SDS na prevenção da calcificação em pericárdio bovino fixado em glutaraldeído: estudo em ratos / Effect of SDS-based decelullarization in the prevention of calcification in glutaraldehyde-preserved bovine pericardium: study in rats

OBJETIVO: Avaliar a descelularização com SDS como tratamento anticalcificante em pericárdio bovino fixado em glutaraldeído. MÉTODOS: Peças de 0,5 cm² foram implantadas em modelo subcutâneo de 18 ratos por até 90 dias. Foram formados quatro grupos: grupo GDA: pericárdio fixado em glutaraldeído 0,5% (GDA), grupo GDA-GL: pericárdio fixado em GDA + ácido glutâmico (GL) 0,2%, grupo D-GDA: pericárdio descelularizado (D) com SDS 0,1% e fixado em GDA e grupo D-GDA-GL: pericárdio descelularizado + GDA + ácido glutâmico 0,2%. Cada animal recebeu enxertos dos quatro grupos. Os explantes foram realizados com 45 e 90 dias. As avaliações foram: análise histológica com as colorações hematoxilina-eosina e alizarina-red, análise morfométrica e quantificação de cálcio por espectrometria de absorção atômica. RESULTADOS: O padrão de infiltrado inflamatório foi o mesmo nos quatro grupos, sendo mais intenso nos grupos GDA e GDA-GL aos 45 dias, ficando mais evidente aos 90 dias. O conteúdo de cálcio aos 45 dias foi de 32,52 ± 3,19 µg/ mg no grupo GDA; 22,12 ± 3,87 µg/mg no grupo GDA-GL; 1,06 ± 0,38 µg/mg no grupo D-GDA e 3,99 ± 5,78 µg/mg no grupo D-GDA-GL (P< 0,001). Aos 90 dias, foi de 65,91 ± 24,67 µg/mg no grupo GDA; 38,37 ± 13,79 µg/mg no grupo GDA-GL; 1,24 ± 0,99 µg/mg no grupo D-GDA e 30,54 ± 8,21 µg/mg no grupo D-GDA-GL (P< 0,001). O grupo D-GDA foi o único que não apresentou progressão da calcificação de 45 para 90 dias (P=0,314). CONCLUSÃO: A descelularização com SDS reduziu o processo inflamatório e inibiu a calcificação em pericárdio bovino implantado em modelo subcutâneo de ratos até 90 dias.

OBJECTIVE: The aim of study was to investigate the SDS-based decellularization process as an anticalcification method in glutaraldehyde-preserved bovine pericardium in subcutaneous rat model. METHODS: Pericardium samples with 0.5 cm² area were divide in four groups: group GDA: 0.5% glutaraldehydepreserved pericardium (GDA); group GDA-GL: GDA + 0.2% glutamic acid (GL); group D-GDA: decellularized (D) pericardium with 0.1% SDS + GDA and group D-GDA-GL: decellularized pericardium + GDA + 0.2% glutamic acid. After this samples were implanted in 18 rats in subcutaneous position till 90 days. Each animal received samples of the four groups. The explants were performed at 45 and 90 days. The explants were subjected to histology in glass slides stained with hematoxilin-eosin and alizarin red, morphometry evaluation and the calcium content was measured by flame atomic absorption spectrometry. RESULTS: The inflammatory infiltrate was the same in all groups, however more intense in GDA and GDA-GL groups in 45 days, increasing at 90 days. The calcium contents for 45 days were: 32.52 ± 3.19 µg/mg in GDA group; 22.12 ± 3.87 µg/ mg in GDA-GL group; 1.06 ± 0.38 µg/mg in D-GDA group and 3.99 ± 5.78 µg/mg in D-GDA-GL (P< 0.001). For 90 days were 65.91 ± 24.67 µg/mg in GDA group; 38.37 ± 13.79 µg/mg in GDA-GL group; 1.24 ± 0.99 µg/mg in D-GDA group and 30.54 ± 8.21 µg/mg in D-GDA-GL (P< 0.001). Only D-GDA did not show increase rates of calcium at 45 to 90 days (P=0.314). CONCLUSION: SDS-based decellularization process reduced the inflammatory intensity and calcification in bovine pericardium in subcutaneous rat model for 90 days.

Can formaldehyde exposure induce histopathologic and morphometric changes on rat kidney? / ¿La exposición al formaldehído puede inducir cambios histopatológicos y morfométricos sobre el riñón de rata?

Formaldehyde is used traditionally for fixing the cadaver, and vaporized during dissection and practical studying on cadaver. This study was designed to determine the histopathologic and morphometric changes of rat kidney while all of the experiments were exposed to formaldehyde for 18 weeks. 28 male albino Wistar rats were divided into the following three experimental groups (E1: 2hrs/d, 2d/w; E2: 2hrs/d, 4d/w; E3: 4hrs/d, 4d/w) and one control group (C). when the exposure period was expired the animals were anaesthetized with chloroform. After cervical dislocation, the abdomen was dissected and the kidneys were taken. The kidney specimens were sectioned and stained with Haematoxylin and Eosin technique for histologic and morphometric study. Data were obtained from an Olympus light microscope and the analyzed with spss (version 11.5) and ANOVA test. In all histopathology sections of groups E1, E2 and E3, these similar changes were seen: mild glumerolar congestion, focal congestion, and vacuolar degeneration of tubular cells. There were no evidences of inflammatory cells infiltration or fibrotic changes of interstitial tissue. Only mild, non-specific congestion was seen in cortical vessels. Also there were not any abnormalities in the staining of nucleus and cytoplasm. According to Morphometric study, Mean +/- SD of glomerulus's area in control, E1, E2 and E3 group were 10802.66 +/- 1038.18, 10759.50 +/- 1971.88, 10434.73 +/- 1763.76 and 10077.64 +/- 2068.78 micrometer, respectively. Mean +/- SD inner proximal tubule diameter in control, E1, E2 and E3 group were 16.16 +/- 2.49, 16.92 +/- 2.90, 16.31 +/-2.79 and15.66 +/-4.11 µm, respectively. Mean +/- SD of inner distal tubule diameter in control, E1, E2 and E3 group were 15.96 +/- 4.47, 16.20 +/- 1.66, 16.96 +/- 1.63 and17.45 +/- 3.26 µm, respectively. These differences were not significant between cases and control. This study showed that formaldehyde inhalation in 1.5 ppm can not make specific...

El formaldehído se utiliza tradicionalmente para la fijación de cadáveres y vaporiza durante la disección y los estudios prácticos en el cadáver. Este estudio fue diseñado para determinar los cambios histopatológicos y morfométricos de riñones de ratas expuestos al formaldehído, durante 18 semanas. 28 ratas albinas Wistar se dividieron en tres grupos experimentales (E1: 2 h/d, 2d/s, E2: 2 h/d, 4d/s; E3: 4 h/d, 4d/s) y un grupo control (C). Cuando el período de exposición se cumplió los animales fueron anestesiados con cloroformo. Después de la dislocación cervical, el abdomen fue disecado y se obtuvieron los riñones. Los especímenes de riñon fueron seccionados y teñidos con técnica hematoxilina y eosina para el estudio histológico y morfométrico. Los datos fueron obtenidos con un microscopio óptico Olympus, el análisis con el software SPSS (versión 11.5) y la prueba de ANOVA. En todas las secciones histopatológicas de los grupos E1, E2 y E3, se observaron cambios similares: congestión glumerular leve, congestión focal y degeneración vacuolar de las células tubulares. No hubo evidencias de la infiltración de células inflamatorias o cambios de tejido fibrótico intersticial. Sólo una leve congestión no específica fue observada en vasos corticales. Además no hubo anormalidad en la tinción del núcleo y el citoplasma. De acuerdo al estudio morfométrico, la Media +/- DS del área glomerular en los grupos control, E1, E2 y E3 fue 10802,66 +/- 1038,18, 10759,50 +/- 1971,88, 10434,73 +/-1763,76 y 10077,64 +/- 2068,78 µm, respectivamente. La Media +/- DS del diámetro interno del túbulo proximal en los grupos control, E1, E2 y E3 fueron 16,16 +/- 2,49, 16,92 +/- 2,90, 16,31 +/- 2,79 y 15,66 +/- 4,11 µm, respectivamente. La Media +/- SD del diámetro interno de los túbulos distales en los grupos control, E1, E2 y E3 fueron 15,96 +/- 4,47, 16,20 +/- 1,66, 16,96 +/- 1,63 y 17,45 +/- 3,26 µm, respectivamente. Estas diferencias no fueron significativas entre los casos...

Proliferating cell nuclear antigen (PCNA), p53 and MDM2 expression in Hodgkins disease

CONTEXT AND OBJECTIVE: Tumor cells in Hodgkins disease (HD) express cell proliferation markers that are evaluated according to the oncogenes involved or the expression of their proteins. Correlations between the protein expression grade and clinical data are now important for disease prognosis. DESIGN AND SETTING: This was a retrospective analysis on proliferating cell nuclear antigen (PCNA), p53 and MDM2 (murine double minute-2) expression using immunohistochemistry, on formalin-fixed, paraffin-embedded tissues from diagnostic biopsies on 51 patients with HD. The study was conducted at the Division of Hematology and Transfusion Medicine, Hospital São Paulo, Universidade Federal de São Paulo. METHODS: Antigen expression was evaluated as the proportions of positive Hodgkin and Reed-Sternberg (HRS) cells and reactive lymphocytes (L), which were compared using Spearman correlation coefficients. The Friedman test was used for comparisons between the markers. The Pearson test was used to investigate associations between marker expression and clinical and laboratory parameters, marrow involvement, complete remission (CR) and overall survival (OS) rates. RESULTS: There was overexpression of antigen proteins in HRS, in relation to L (p < 0.001). In HRS, MDM2 was higher than p53 and PCNA (p < 0.003), while the latter two were equivalent. In L, p53 was lower than MDM2 and PCNA (p < 0.001), while the latter two were equivalent. There was no relationship between protein expression and clinical and laboratory variables or outcome. CONCLUSIONS: PCNA, p53 and MDM2 are tumor markers for HD, but showed no clinical or prognostic significance in our analysis.

CONTEXTO E OBJETIVO: As células tumorais da doença de Hodgkin (HD) são positivas para marcadores de proliferação celular que são analisados por seus genes e respectivas proteínas. A correlação entre a expressão destas proteínas e os parâmetros clínico-laboratoriais são, no momento, de importância para o prognóstico da doença. TIPO DE ESTUDO E LOCAL: Estudo retrospectivo da expressão do antígeno de proliferação celular (PCNA) e da p53 e MDM2 em tecidos obtidos ao diagnóstico, fixados por formol, embebidos em parafina de 51 pacientes com HD. O trabalho foi realizado na Divisão de Hematologia e Transfusão, Hospital São Paulo, Universidade Federal de São Paulo. MÉTODOS: As expressões antigênicas foram analisadas através da proporção de células de Hodgkin e células de Reed Sternberg (HRS) e linfócitos reacionais (L) positivos. A intensidade de expressão de cada proteína foi comparada entre L e HRS através do coeficiente de Spearman. A comparação da PCNA, p53 e MDM2 em L e HRS se fez pelo teste de Fiedman. As correlações entre variáveis clínico-laboratoriais, comprometimento da medula óssea, taxas de sobrevida geral e remissão clínica com as proteínas em HRS se fizeram pelo coeficiente de Pearson. RESULTADOS: Houve superexpressão das três proteínas em células HRS comparadas aos L (p < 0,001). Nas células HRS, a MDM2 foi maior que a p53 e a PCNA (p < 0,003), que foram equivalentes. Nos L, a p53 foi menor que a MDM2 e a PCNA (p < 0,001), que foram equivalentes Não houve relação entre as expressões das proteínas com as variáveis clínico-laboratoriais e sobrevida. CONCLUSÕES: PCNA, p53 e MDM2 são marcadores tumorais na HD, porém não mostraram significado clínico-prognóstico em nossa análise.

Evaluation of fixative solutions for ultrastructural analysis of brown spider Loxosceles intermedia (araneae: sicariidae) tissues

In view of the widely varying compositions of fixative solutions used for studying spiders, five different fixative formulas were tested for fixing male brown-spider (Loxosceles intermedia) gonad tissues. The brown spider represents a public health problem in Curitiba (Paraná State, Brazil). Morphological study of its gonads may aid in understanding the reproductive strategies of this species, and possibly in developing a reproduction control program. The fixatives tested contained glutaraldehyde alone or combined with paraformaldehyde, and the buffers cacodylate or phosphate, with or without the addition of sucrose or sodium chloride as osmolytes. Those containing 2.5 percent glutaraldehyde and 2 percent paraformaldehyde in 100 mM phosphate buffer with 200 mM sucrose, or in 200 mM sodium cacodylate, satisfactorily preserved mitochondria, the Golgi apparatus, and the membranes in general. These formulas were nearly isosmotic (439 mOsm/kg H2O and 455 mOsm/kg H2O respectively) to brown spider hemolymph (478 mOsm/kg H2O). With respective to the fixative agents, a glutaraldehyde-paraformaldehyde combination resulted in optimal fixation of Loxosceles intermedia cells. For other species of spiders, hemolymph osmolality should be considered, but the fixative formulas cited above would also probably yield good results.

Dada a variabilidade na composição de soluções fixadoras utilizadas em aranhas, cinco diferentes fixadores foram elaborados para a análise ultra-estrutural dos tecidos da aranha marrom Loxosceles intermedia. A aranha marrom representa um problema de saúde pública na cidade de Curitiba, e o estudo morfológico de suas gônadas pode auxiliar na compreensão de suas estratégias reprodutivas e, possivelmente, no desenvolvimento de um programa de controle da sua população. As fórmulas usadas continham glutaraldeído com ou sem paraformaldeído, tampão cacodilato ou fosfato, e NaCl ou sacarose como osmólitos. As soluções fixadoras compostas por 2.5 por cento glutaraldeído e 2 por cento paraformaldeído, em tampão fosfato com adição de sacarose ou em 200 mM cacodilato de sódio, preservaram bem estruturas como mitocôndrias, aparelho de Golgi e membranas em geral. Os tampões são praticamente isosmóticos (439 mOsm/kg H2O e 455 mOsm/kg H2O, respectivamente) à hemolinfa da aranha marrom (478 mOsm/kg H2O). Ainda, com relação aos agentes fixadores, a combinação do glutaraldeído e paraformaldeído levou a uma melhor preservação das células. Para outras espécies de aranhas, a osmolalidade da hemolinfa deve ser medida e considerada, mas as fórmulas acima citadas podem ser testadas, com chance de sucesso.

Colchicine binding to tubulin from brain homogenates in the presence of the sugars, glycols and metal ions: effect of nickel ions on the tubulin solubility

The fact that glycerol preserves microtubules from depolymerizing in vitro, and that some ions such as Ca(II) and Mg(II), regulate the assembly-disassembly process of these structures, induced us to study the effect of several sugars, glycols and metal ions on solubility and colchicine affinity of tubulin in rat brain homogenates, and of purified microtubular protein. Inhibition of colchicine binding was significant with glycerol, polyethylene glycol 1000 (PEG-2) and the ions A1(III), Co(II), Ni(II), while compounds structurally related to glycero (glucose and sucrose) did not inhibition it. Mannitol, instead, increased the activity a 47% over control. Apparently the presence of some compounds in brain homogenates [PEG-2 (1000) and NI (II)] favored tubulin sedimentation when these latterwere centrifuged at 100,000 x g for 150 min at 20 degrees C, but the form in which tubulin becomes aggregated in the pellet is unknown. Nickel ion madeinsoluble microtubular protein of homogenates and the purified one by more than 90% without causing significant inhibition of the colchicine binding. The sediment containing nickel-treated two cycles purified microtubular protein observed with the electron microscope did not present microtubules, but it revealed the presence of irregular, wavy and streteched structures, but it revealed the presence of irregular, wavy and stretched structures bearing highly dense dotted material. The sediments became soluble in phosphate-glutamate buffer (pH 6.8) and, when incubated in polymerizing conditions, gave rise to microtubules undistinguishable from those prepared with untreated purified protein