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.

Preservation of histology by phenol-based fixative: mini review of recent findings / Preservación de la histología mediante un fijador basado en fenol: una mini revisión de los hallazgos recientes

SUMMARY: In surgical and anatomical training, use of cadaver remains the most ideal technique. Standard formaldehyde solution preserves cadaveric tissues for an extended period comparing to the unfixed tissues. However, it fails to retain the natural texture, color, and biomechanical features. Phenol based soft embalming methods were developed to maintain these properties, while simultaneously decreasing the biohazard risk. Soft embalming techniques have made the bodies more 'lifelike' and wellfitted for training. Though phenol fixation displays rewarding morphological maintenance, we have scanty evidences on the histological preservation. This mini review primarily discussed the latest reports regarding the effect of phenol-based fixation on the tissue histology. Published literatures revealed phenol-based fixation displayed comparable histological preservation to that ofgold standard paraformaldehyde-based solution. It was concluded that phenol-based solution is an excellent fixative used to preserve tissues for microscopic analysis.

RESUMEN: En el entrenamiento quirúrgico y anatómico, el uso de cadáveres sigue siendo la técnica más ideal. La solución estándar de formaldehído conserva los tejidos cadavéricos durante un período prolongado en comparación con los tejidos no fijados. Sin embargo, no conserva la textura, el color y las características biomecánicas naturales. Se desarrollaron métodos de embalsamamiento blando a base de fenol para mantener estas propiedades y, al mismo tiempo, disminuir el riesgo biológico. Las técnicas de embalsamamiento suaves han hecho que los cuerpos sean más "realistas" y estén mejor preparados para la enseñanza. A pesar que la fijación de fenol muestra un buen mantenimiento morfológico, existe evidencia escasa sobre la preservación histológica. Esta mini revisión se refirió principalmente a los últimos informes sobre el efecto de la fijación en base de fenol en la histología del tejido. La literatura publicada reveló que la fijación a base de fenol mostró una preservación histológica comparable a la de la solución a base de paraformaldehído. Se concluyó que la solución a base de fenol es un excelente fijador utilizado para preservar tejidos para análisis microscópico.

Use of Chilean conservative fixative solution in veterinary anatomical parts / Uso de la solución conservadora chilena en piezas anatómicas veterinarias

SUMMARY: The conservation of anatomical pieces goes back five thousand years, beyond ancient Egypt for political and religious purposes. In the 12th century, with the appearance of the University, new techniques were implemented as the corpse became a necessary tool for teaching. During the last two centuries, since the year of its discovery in 1867, formaldehyde has been used as the main source for conservation. However, several years ago, anatomy laboratories around the world have resorted to new techniques and mixtures in order to reduce its use and its reported negative effects on health, such as cancer. This work shows the obtaining of anatomical pieces, by means of the Chilean Conservative Fixative Solution (SFCCh), its process, analysis and experience are described in the following pages.

RESUMEN: La conservación de piezas anatómicas se remonta a cinco mil años, más allá del antiguo Egipto por motivos políticos y religiosos. En el siglo XII, con la aparición de la Universidad, se implementaron nuevas técnicas ya que el cadáver se convirtió en una herramienta necesaria para la enseñanza. Durante los dos últimos siglos, desde el año de su descubrimiento en 1867, el formaldehído se ha utilizado como principal fuente de conservación. Sin embargo, hace varios años, los laboratorios de anatomía de todo el mundo han recurrido a nuevas técnicas y mezclas con el fin de reducir su uso y sus reportados efectos negativos en la salud, como el cáncer. En este trabajo se muestra la obtención de piezas anatómicas, mediante la Solución Fijadora Conservadora Chilena (SFCCh), su proceso, análisis y experiencia se describen en las siguientes páginas.

Alcohol-based fixatives can better preserve tissue morphology than formalin / Fijadores a base de alcohol pueden preservar mejor la morfología tisular que la formalina

SUMMARY: Fixation is a crucial step in processing of tissue specimen for preservation of cellular architecture and composition of cells. Alcohol-based fixatives are considered some of the most promising alternatives to formalin. We evaluated the performance of alcohol-based fixatives (EthMeth and methacarn) and formalin as a comparator fixative in the research laboratory. Following 24 hours of fixation, tissue morphology and cellular details of the liver, spleen and brain (cerebral cortex) were evaluated. Morphological characteristics were evaluated by gross observations and analyzing cellular details, tissue architecture and overall staining characteristics (Hematoxylin and Eosin). EthMeth and methacarn fixation gave generally comparable and satisfactory results on the tissue morphology and subsequent identification of tissue characteristics. Particularly, tissues were well preserved and all nuclear as well as cytoplasmic details were clearly visible. However, formalin fixed tissues showed some peculiarity such as improper fixation, mild shrinkage, and alterations of tissue components. These results confirm that alcohol-based fixation is the superior alternative to formalin for preservation of tissue morphology. However, it is required to standardize the formalin-free methods and harmonize diagnosis in the laboratory worldwide.

RESUMEN: La fijación es un paso crucial en el procesamiento de muestras de tejido para preservar la arquitectura celular y la composición de las células. Los fijadores a base de alcohol se consideran algunas de las alternativas más prometedoras a la formalina. Evaluamos el rendimiento de los fijadores a base de alcohol (EthMeth y metacarn) y formalina como fijador comparativo en el laboratorio de investigación. Después de 24 horas de fijación, se observó la morfología del tejido y los detalles celulares del hígado, bazo y corteza cerebral. Se evaluaron las características morfológicas mediante observaciones generales y analizando detalles celulares, arquitectura de tejidos y características generales de tinción (hematoxilina y eosina). La fijación de EthMeth y metacarn dio resultados generalmente comparables y satisfactorios en la morfología del tejido y la posterior identificación de las características del mismo. Particularmente, los tejidos estaban bien conservados y todos los detalles nucleares y citoplasmáticos eran claramente visibles. Sin embargo, los tejidos fijados con formalina mostraron cierta peculiaridad, tal como una fijación inadecuada, la contracción leve y alteraciones de los componentes del tejido. Estos resultados confirman que la fijación a base de alcohol es la mejor alternativa a la formalina, para preservar la morfología del tejido. Sin embargo, es necesario estandarizar los métodos sin formalina y armonizar el diagnóstico en los laboratorios.

Rabbit growth plate morphology in temporary bilateral blocking / Morfología de la placa de crecimiento de conejos durante bloqueo bilateral temporario

Blocking of the growth plate (GP) using plates with screws (tension band plating) is a modern method used to correct deformities and moderate leg length discrepancy in growing children. Determining the duration of temporary bilateral blocking without the occurrence of irreversible changes of GP is of paramount importance important. Methods: Two-month-old Californian breed male rabbits (n=30) were exposed to bilateral blocking of the distal GP of the right femur locking plates with screws for 3, 5, and 7 weeks. The fixators were removed after 5 and 7 weeks in 18 rabbits and 3 weeks after that, animals were sacri!ced. The contralateral limb was used as a control. Histological, histomorphometric, and X-ray analyses were performed. Results: During GP blocking, its height gradually decreased. This decreased was more pronounced after 7 weeks. Destructive changes progressed with an increase in the blocking duration. Three weeks after discontinuation of the bilateral blocking that lasted 5 weeks, the height of the GP signi!cantly increased 1.2 times on the lateral side and 1.9 times on the medial side (p<0.001) compared to the control. When blocking was discontinued after 7 weeks, the structure of the GP was partially restored after 3 weeks, the height of GP signi!cantly increased 1.2 times on the lateral side, and 1.07 times on the medial side (p<0.01) compared to the control. Conclusion: Restoration of the structuralfunctional features of the GP after the removal of the plates depends on the duration of temporary bilateral blocking, which must be taken into account in the clinical setting. (AU)

El bloqueo de la placa de crecimiento (PC) utilizando placas con tornillos (banda de tensión) es un método moderno utilizado para corregir deformidades y alteraciones moderadas en la longitud de las piernas en niños en crecimiento. Es de suma importancia determinar cuál debe ser la duración del bloqueo bilateral temporal sin que ocurran cambios irreversibles en la PC. Métodos: Conejos machos de raza californiana de dos meses de edad (n = 30) fueron expuestos al bloqueo bilateral de la PC distal colocando placas del fémur derecho con tornillos durante 3, 5 y 7 semanas. Los fijadores fueron retirados después de 5 y 7 semanas en 18 de los conejos, y 3 semanas después los animales fueron sacrificados. La extremidad contralateral se utilizó como control. Se realizaron análisis histológicos, histomorfométricos y de rayos X. Resultados: Durante el bloqueo de la PC, su altura disminuyó gradualmente. Esta disminución fue más pronunciada después de 7 semanas. Los cambios destructivos se incrementaron a medida aumentaba la duración del bloqueo. Tres semanas después de la interrupción del bloqueo bilateral que duró 5 semanas, la altura de la PC aumentó significativamente 1.2 veces en el lado lateral y 1.9 veces en el lado medial (p <0.001) en comparación con el control. Conclusión: La restauración de las características funcionales estructurales de la PC después de la extracción de las placas depende de la duración del bloqueo bilateral temporal, lo que debería tenerse en cuenta en el tratamiento clínico de estas alteraciones. (AU)

Preparación de hemisferios cerebrales para disección de tractos / Preparation of cerebral hemispheres for tracts dissection

Desde la antigüedad se han desarrollado técnicas para el estudio del cerebro con fines didácticos o neuroquirúrgicos. Hacia 1934 Josef Klingler desarrolla una técnica de preparación de hemisferios cerebrales que basada en la fijación con formalina y el congelamiento para aislar los tractos cerebrales. El objetivo de la presente artículo ha sido analizar los métodos de preparación utilizados para la disección de tractos en cerebros humanos y de animales. Se realizó una revisión de la literatura en las bases de datos Web of Science, Scopus, Pubmed, Medline y Scielo, utilizando como descriptores: Disección, Cerebro, Tracto, con el operador booleano "AND" entre ellos, en los idiomas inglés y español, hasta junio de 2018. Fueron seleccionados 26 documentos, para el análisis se determinaron las variables: espécimen, número de hemisferios cerebrales, concentración de formalina, tiempo de fijación, temperatura, tiempo de congelamiento y tractos identificados. En la literatura seleccionada, un total de 410 hemisferios cerebrales fueron analizados, 372 de humanos y 38 de animales; 330 fueron conservados en formalina al 10 %, 20 en formalina al 5 % y el resto en otras concentraciones. El tiempo de fijación fue variable entre 10 y 180 días, así como la temperatura y tiempo de congelación (-10 ºC y -20 ºC, entre 8 y 30 días). En todos los casos se reportó que, en su totalidad o parcialmente, los fascículos cerebrales de asociación fueron aislados. En la preparación de hemisferios cerebrales para disección de tractos, Ludwig & Klingler (1956) recomiendan que en la fijación de los especímenes se utilice formalina al 5 %, sin embargo, el 80 % de los hemisferios utilizados fueron fijados en formalina al 10%, y en esta concentración, el tiempo de fijación, temperatura y tiempo de congelación fue variable, lográndose, en todos los casos analizados, la disección parcial o total de los tractos.

Since ancient times, techniques for the study of the brain have been developed for didactic or neurosurgical purposes. By 1934, Josef Klingler developed a cerebral hemisphere preparation technique based on formalin fixation and freezing to isolate the cerebral tracts. The aim of this article was to analyze the preparation methods used for tracts dissection in human and animal brains. A review of the literature using Web of Science, Scopus, Pubmed, Medline and Scielo databases, with the following descriptors: Dissection, Brain, Tract, with the boolean operator "AND" among them, also in spanish, until June 2018. Twenty-six documents were selected, and we analized the following variables: specimen, number of cerebral hemispheres, formalin concentration, fixing time, temperature, freezing time and tracts. In the selected literature, a total of 410 cerebral hemispheres were analyzed, 372 from humans and 38 from animals; 330 were preserved in 10 % formalin, 20 in 5 % formalin and the rest in other concentrations. The fixation time was variable between 10 and 180 days, as well as the temperature and freezing time (-10 ºC and -20 ºC, between 8 and 30 days). In all cases it was reported that, in whole or in part, the cerebral fascicles of association were isolated. In the preparation of cerebral hemispheres for dissection of tracts, Klingler recommend that 5 % formalin for the fixation of specimens; however, 80 % of the hemispheres used were fixed in 10 % formalin, and in this concentration, the time of fixation, temperature and time of freezing was variable, achieving, in all the cases analyzed, the partial or total dissection of the tracts.

Femoral Periprosthetic Fractures after Total Knee Arthroplasty: New Surgically Oriented Classification with a Review of Current Treatments

PURPOSE: As the number of total knee arthroplasties (TKAs) increases, the incidence of femoral periprosthetic fractures after TKA is also increasing. This review aimed to suggest a new surgically oriented classification system for femoral periprosthetic fractures. METHODS: We investigated the classifications, and current treatment trends for femoral periprosthetic fractures after TKA by means of a thorough review of the relevant literature. RESULTS: Numerous studies reported good results of surgical treatment with modern fixatives including locking compression plates and retrograde intramedullary nails. However, few classifications of femoral periprosthetic fractures reflect the recent developments in surgical treatment. CONCLUSIONS: We recommend that surgical management be considered the first-line treatment for femoral periprosthetic fractures after TKA. Our new classification will help in deciding the surgical treatment option for femoral periprosthetic fractures after TKA.

Avaliação de diferentes fixadores na preservação das características histológicas de pele de orelha de cães / Evaluation of different fixatives for histological studies in canine ear skin

Os fixadores biológicos desempenham um papel importante na qualidade final da histologia. Na rotina veterinária, a biópsia de pele é um procedimento comum e a escolha do fixador é primordial para resultado final adequado. Os fixadores mais usados são à base de formalina, ainda que sejam tóxicos, cancerígenos, de baixa penetração e de fixação lenta. Mesmo assim, não existe um fixador ideal que substitua as suas qualidades. O objetivo deste trabalho foi avaliar qualitativamente a preservação das características histológicas de pele de cão utilizando diferentes fixadores de tecidos incluídos em parafina, cortados e corados pela hematoxilina-eosina. Utilizou-se uma caneta Punch de 4 milímetros para coletar amostras de pele de orelha em seis cadáveres de cães. Após coleta, os tecidos foram fixados em: (1) Bouin, durante seis horas; (2) Carnoy, durante quatro horas; (3) formaldeído tamponado 10% durante 24 horas, todos sob refrigeração (4ºC). Posteriormente, os tecidos foram processados, cortados e corados em hematoxilina e eosina. As lâminas foram avaliadas, às cegas, por quatro patologistas diferentes, que consideraram aspectos qualitativos a seguir: (1) qualidade da coloração; (2) preservação das características histológicas; e (3) preservação dos limites citoplasmáticos utilizando a escala de LIKERT de pontuação para cada lâmina. O fixador com a maior média de pontuação em todos os itens foi o formol tamponado com 3,76 pontos, seguido pelo Bouin (3,39) e pelo Carnoy (2,52). O formol pode trazer riscos à saúde do profissional que rotineiramente o manuseia, portanto se faz necessária a busca por fixadores com as mesmas qualidades, mas menos nocivos à saúde.(AU)

The biological fixatives have an important role in the final histology quality. In veterinary, routine skin biopsy is a common procedure and the choice of fixative is essential for the final result. The most common fixative is Formalin, even though it is toxic, carcinogenic, and has low and slow penetration. Still, there isn't a fixer which can replace the qualities of formalin. The aim of this study was to evaluate qualitatively the preservation of the histological features of dog skin using different tissue fixative embedded in paraffin, sectioned and stained with hematoxylin - eosin. We used a 4 mm punch pen to collect ear skin samples in six dog cadavers. After collection, the tissues were fixed in: (1) Bouin for 6 hours; (2) Carnoy for 4 hours; (3) 10% buffered formaldehyde for 24 hours, all under refrigeration (4 ° C). The tissues were then processed, sectioned and stained with hematoxylin and eosin. The slides were evaluated blindly by four different pathologists who considered the qualitative aspects below: (1) quality of coloring; (2) preservation of the histological characteristics; (3) preservation of cytoplasmic boundaries using a Likert scale score for each blade. The fixative with the highest mean score on all items was buffered formalin with 3.76 points followed by Bouin (3.39) and Carnoy (2.52). Formaldehyde can bring health a risk of professional routine handling, so it is necessary to search for a biological fixative with the same qualities being less harmful to health.(AU)

Low impact to fixed cell processing aiming transmission electron microscopy

In cell culture, cell structures suffer strong impact due to centrifugation during processing for electron microscope observation. In order to minimise this effect, a new protocol was successfully developed. Using conventional reagents and equipments, it took over one week, but cell compression was reduced to none or the lowest deformation possible.

Comparison of Quality of Life between Before and After Orthopaedic Implant Removal Surgery

PURPOSE: The purpose of this study is to determine whether or not a patient's results are improved after removal of an internal fixative from a patient with no related symptoms. MATERIALS AND METHODS: This prospective study included 87 patients who agreed to participate in the study and satisfied the criteria for selection and exclusion of patients who underwent the operation for removal of internal fixative due to broken bones from March 1st, 2004 to December 31st, 2011 at Daegu Catholic University Medical Center. The average replication period was 27 months (12-64 months) and the average age at the time of the operation for removal was 41.5 years (21-75 years) for 55 males and 32 females. The quality of life for all patients was evaluated using Short Form 36 (SF-36) surveys before the operation for removal and after a minimum of one year. RESULTS: After an orthopedic operation for removal of internal fixative, physical health status showed statistically significant improvement (p=0.001); however mental health status did not (p=0.411). A satisfaction test for the subjective surgery written by patients indicated an improvement of subjective health status in 52.9% after the surgery for removal but with no difference in 29.9% compared to preoperation. CONCLUSION: In case of an operation for removal of internal fixative for patients with no related symptoms with internal fixatives used for treatment of fractures showing agglutination opinions, an improvement was observed in physical health status, not in mental health status. When surgery for removal of internal fixative is performed for patients without related symptoms, consideration that subjective satisfaction of patients shows an improvement only in 52.9% will be helpful.

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. .

Comprehensiveness and programmatic vulnerability to stds/hiv/aids in primary care / La integralidad y vulnerabilidad programática de las ets/vih/sida en la atención básica / A integralidade e a vulnerabilidade programática às DST/HIV/AIDS na Atenção Básica




This study aimed to identify programmatic vulnerability to STDs/HIV/AIDS in primary health centers (PHCs). This is a descrip - tive and quantitative study carried out in the city of São Paulo. An online survey was applied (FormSUS platform), involving administrators from 442 PHCs in the city, with responses received from 328 of them (74.2%), of which 53.6% were nurses. At - tention was raised in relation to program - matic vulnerability in the PHCs regarding certain items of infrastructure, prevention, treatment, prenatal care and integration among services on STDs/HIV/AIDS care. It was concluded that in order to reach comprehensiveness of actions for HIV/ AIDS in primary health care, it is necessary to consider programmatic vulnerability, in addition to more investment and reor - ganization of services in a dialogue with the stakeholders (users, multidisciplinary teams, and managers, among others).


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Objetivo Fue identificar la vulnerabilidad programática de las Unidades Básicas de Salud con la atención a las ETS/VIH/SIDA. Método Es un estudio descriptivo con un abordaje cuantitativo llevado a cabo en el Municipio de San Pablo. Fue utilizado un formulario online (el FormSUS) con los gerentes de las 442 Unidades Básicas de Salud del Municipio de San Pablo. Participaran en el estudio 74.2% de los gerentes, estos 53.6% eran enfermeros. Resultados Se destaca la vulnerabilidad programática de las Unidades Básicas de Salud en relación a algunos elementos de la infraestructura, acciones de prevención, tratamiento, prenatal y la integración entre los servicios en la atención a las ETS/VIH/SIDA. Conclusión La construcción de tales marcadores constituye un instrumento, presentado en otro artículo, el cual puede ayudar a apoyar la captura de vulnerabilidades de las mujeres en relación a las ETS/VIH en el contexto de los servicios de Atención Primaria de Salud. Los marcadores constituyen importante herramienta para operacionalizar el concepto de vulnerabilidad en la Atención Primaria. Además, promueven procesos de trabajo inter e multidisciplinar e inter e multisectorial. La propuesta de un instrumento basado en dichos marcadores puede apoyar la captura de la vulnerabilidad de las mujeres en relación a las ETS/VIH. .

Objetivo Identificar a vulnerabilidade programática às DST/HIV/aids na Atenção Básica para o enfrentamento do HIV/Aids. Método Estudo descritivo, com abordagem quantitativa, realizado no Município de São Paulo (MSP). Utilizou-se formulário online (FormSUS), com gerentes das 442 Unidades Básicas de Saúde (UBS) do MSP. Participaram do estudo 74,2% gerentes, dos quais 53,6% eram enfermeiros. Resultados Destaca-se a vulnerabilidade programática nas UBS com relação a alguns itens de infraestrutura, ações de prevenção, de tratamento, no pré-natal e de integração entre os serviços na atenção às DST/HIV/aids. Conclusão Para a efetivação da integralidade no enfrentamento do HIV/aids na Atenção Básica é necessário atentar para a vulnerabilidade programática, além de mais investimentos e reorganização dos serviços, num diálogo com os atores sociais envolvidos (usuários, equipe multiprofissional, gerentes, gestores, entre outros).


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Epidermal growth factor receptor gene expression evaluation in colorectal cancer patients.

Background: Colorectal cancer is one of the most common causes of death in the world and third and fourth most common cancer among men and women in Iran respectively. Epidermal growth factor receptor (EGFR) is a tyrosine kinase receptor that shows over expression in epithelial tumors and regulates important processes in tumorigenesis. Incidence and characteristics of colorectal cancer are based on the geographic region and race. Aim: In this research work, the over expression of EGFR in formalin fixed paraffin-embedded (FFPE) colorectal cancer tumor tissue of patients was studied. Materials and Methods: Fifteen FFPE colorectal cancer tumor tissues (10 women and 5 men; 25-65 years old and stage IV) and 15 non-patients (nine women and six men; 25-65 years old) that were collected during 2006-2012. EGFR gene expression level was analyzed by real-time quantitative reverse transcriptase polymerase chain reaction (PCR). All PCR reactions were performed in triplicate for both target gene and internal control (18s ribosomal ribonucleic acid) with the 2−ΔΔCT method. Gene expression differences in patients and controls were evaluated with t-test. Results: The results were showed EGFR gene over expression in 12 (80%) of 15 patients. There was a statistically significant difference in the prevalence of EGFR expression between patients and control (P < 0.05). Conclusion: Our results demonstrated EGFR gene over expression in colorectal cancer tumor tissue compared with controls.

HER2 protein testing in gastric cancer: a retrospective analysis of 1 471 cases during two different periods in a single medical center / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the potential factors in influencing the performance of immunohistochemical testing for HER2 protein in gastric cancers.</p><p><b>METHODS</b>The HER2 protein expression status of 1 471 surgically resected archival gastric cancer cases in Drum Tower Hospital collected during two different periods was retrospectively analyzed. The materials included 957 cases tested during the period from 2007 to 2009 (group 1) and 514 cases from 2012 to 2013 (group 2). The test procedures and results observed during these two periods were compared.</p><p><b>RESULTS</b>The percentages of score 3 HER2 protein expression (14.4%, 74/514 versus 9.5%, 91/957) and score 2 or score 3 HER2 protein expression (27.2%, 140/514 versus 21.7%, 208/957) were both higher in group 2 than in group 1 (P < 0.05). In group 1, the cancer tissue was fixed in 10% formalin, stained manually with HER2 antibody A0485 (Dako) and assessed by different pathologists.In group 2, the tissue was fixed in 10% neutral buffered formalin (pH 7.2), stained using automated immunostaining system (Roche Benchmark XT) with HER2 antibody 4B5 (Ventana) and assessed by a specialized team of pathologists.</p><p><b>CONCLUSION</b>The results of HER2 immunostaining in gastric cancer are influenced by a number of factors including type of fixative, clone number of primary antibody, staining methods and experience of pathologists.</p>

A Comparative Study to Determine the Most Suitable Fixative for Immunocytochemical Staining of Cytospin Slides / 임상검사와정도관리

BACKGROUND: Fixation of cells is a critical procedure that can determine the success of immunocytochemical staining (ICC) of cytospin slides. In this study, we evaluated the efficacy of a number of fixatives to determine the ideal fixative for ICC of cytospin slides. METHODS: Sixteen patients with metastatic neoplasm in the body cavity were enrolled. Cytospin slides were prepared from each patient using 5 different fixatives (cold acetone, 95% ethanol, 1:1 methanol:ethanol, 3.7% formalin, and 3:1 methanol:acetone), and the suitability of each for use with Wright's stain was compared. For 4 of the samples, appropriate ICCs were performed using all 5 fixatives and the results were compared, while for 11 samples, only the first 3 fixatives were used for ICC. RESULTS: Using Wright stain, cold acetone, 95% ethanol, and 1:1 methanol:ethanol fixatives all showed similar efficacy when compared to the conventional methanol fixation method. However, the stain quality using 3.7% formalin or 3:1 methanol:acetone fixatives was poor due to deterioration of cell adhesion and distortion of cell morphology. Using ICC, cold acetone fixative showed stronger and more tumor-specific stainability than the 95% ethanol (decreased stainability in 6 stained slides, false positive staining of histiocytes/neutrophils in 4 stained slides, no staining of CD3 and terminal deoxynucleotidyl transferase [TdT]) and 1:1 methanol:ethanol fixatives (decreased stainability in 3 stained slides, false positive staining of histiocytes/neutrophils in 2 stained slides, no staining of CD3 and TdT). CONCLUSIONS: Cold acetone fixative was the most efficacious among the 5 fixatives tested in this study; therefore, it is the most appropriate fixative in the preparation of cytospin slides for ICC.

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.

Stool sample storage conditions for the preservation of Giardia intestinalis DNA

Stool is chemically complex and the extraction of DNA from stool samples is extremely difficult. Haemoglobin breakdown products, such as bilirubin, bile acids and mineral ions, that are present in the stool samples, can inhibit DNA amplification and cause molecular assays to produce false-negative results. Therefore, stool storage conditions are highly important for the diagnosis of intestinal parasites and other microorganisms through molecular approaches. In the current study, stool samples that were positive for Giardia intestinalis were collected from five different patients. Each sample was stored using one out of six different storage conditions [room temperature (RT), +4ºC, -20ºC, 70% alcohol, 10% formaldehyde or 2.5% potassium dichromate] for DNA extraction procedures at one, two, three and four weeks. A modified QIAamp Stool Mini Kit procedure was used to isolate the DNA from stored samples. After DNA isolation, polymerase chain reaction (PCR) amplification was performed using primers that target the β-giardin gene. A G. intestinalis-specific 384 bp band was obtained from all of the cyst-containing stool samples that were stored at RT, +4ºC and -20ºC and in 70% alcohol and 2.5% potassium dichromate; however, this band was not produced by samples that had been stored in 10% formaldehyde. Moreover, for the stool samples containing trophozoites, the same G. intestinalis-specific band was only obtained from the samples that were stored in 2.5% potassium dichromate for up to one month. As a result, it appears evident that the most suitable storage condition for stool samples to permit the isolation of G. intestinalis DNA is in 2.5% potassium dichromate; under these conditions, stool samples may be stored for one month.

Estudio comparativo entre soluciones conservadoras con y sin formol en placenta humana / Comparative study between conservative solutions with and without formaldehyde in human placenta

Las técnicas de fijación y conservación anatómica son realizadas habitualmente con soluciones que contienen formol, dado su bajo costo. Estas tienen varias desventajas como el olor irritante, rigidez, cambios de coloración de las estructuras, así como toxicidad con potencial cancerígeno, teratogénico y mutagénico para quien lo manipula. Por esto, es deseable utilizar soluciones sin formol. El objetivo de este trabajo fue comparar 2 métodos de conservación cadavérica, uno con formol (solución de Montevideo) y otro sin formol (método de Prives) utilizando la placenta humana como órgano experimental, evaluando sus parámetros macroscópicos. Se utilizaron 46 placentas humanas de partos normales y gestación a término. Las placentas fueron separadas en dos grupos (n=22 y n=24 respectivamente). El primer grupo de placentas fue perfundido con una solución con formol y el segundo grupo en una solución sin formol. Luego ambos grupos fueron sumergidos y mantenidos en sus soluciones respectivas por dos semanas y posteriormente retiradas dejándolas al aire a temperatura ambiente por 2 semanas más. Se analizaron las variables cuantitativas de peso y diámetro en cada una de las piezas, así como las variables cualitativas de consistencia, color, olor y crecimiento de micro/macro organismos. La recopilación de datos fue realizada previo al lavado, a los 14, 21 y 28 días. Los resultados mostraron que las placentas conservadas con el método de Prives presentaron mejor conservación en relación a su diámetro, consistencia, color y menor olor irritante en relación a las placentas tratadas con solución con formol. En ningún caso hubo crecimiento de micro o macroorganismos. En conclusión, emplear soluciones alternativas que sustituyan ventajosamente al formol como la fórmula de Prives conservan mejor las características macroscópicas de las placentas sin generar un olor irritante, deteniendo el proceso de descomposición.

The fixation and conservation techniques of anatomic material are commonly performed with solutions containing formaldehyde, given its low cost. These have several disadvantages such as the irritating odor, stiffness, discoloration of the structures, toxicity, carcinogenic, teratogenic and mutagenic risk for those who are exposed. Therefore it is desirable to use solutions without formaldehyde. The aim of this study was to compare 2 methods of anatomical conservation, one with formalin (Montevideo's solution) and one without formalin (Prives method) using the human placenta as an experimental organ model evaluating its macroscopic parameters. We used 46 human placentas from normal deliveries and term pregnancy. The placentas were separated into two groups (n=22 and n=24 respectively). The first group of placentas was perfused with formaldehyde solution and the second group in a solution without formaldehyde. Then both groups were immersed and maintained in their respective solutions for two weeks and then withdrawn leaving the air at room temperature for 2 weeks. Quantitative variables were analyzed for weight and diameter of each piece, and qualitative variables as consistency, color, odor and growth of micro/macro organisms were evaluated. Data collection was performed prior to washing at 14, 21 and 28 days. The results showed that conserved placentas with Prives method showed better conservation in relation to its diameter, consistency, color and less irritating odor in relation to placentas treated with formalin solution. In no case was growth of micro or macro organism. In conclusion, using advantageously at alternative solutions to formaldehyde, as the formula of Prives method, better preserved macroscopic characteristics of placentas without generating an irritating smell, stopping the decomposition process.

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.

[Evaluation of formalin substitution fixative effects on H and E staining results]

These days, Formalin is considered as a good and cheap fixative, but very harmful for human health. It is one of chemical components which has traditionally been used as a routine fixative in anatomy and pathology laboratories; however, their personnel are still under its harmful effects. In this study, effects of formalin substitution fixatives on H and E staining results were investigated. Three kinds of formalin substitution beside 10% buffered formalin were used in this study. They were named and recognized as B, K, M. Pieces of rabbit liver were placed in dishes which contained each kind of above mentioned fixatives. After 24 hours, tissues were processed and embedded by paraffin followed by sectioning, staining by hematoxylin and eosin. Then, they were studied under light microscope. Overall, preserving of tissue structures and staining results with M fixative were more desirable. In general, two out of the three fixatives, with some advantages and disadvantages rather than formalin, made tissues fixed desirably, and had acceptable results in staining. Thus, they can be known as suitable substitution for formalin