MOHS SURGERY: NARRATIVE REVIEW

Technique widely used in dermatology for the treatment of skin cancer. The historical perspective highlights the evolution of Mohs surgery since its development by Dr. Frederic Mohs in the 1930s, emphasizing its focus on basal cell carcinoma and subsequent renements in instrumentation, anesthesia, and tissue processing methods. Mohs surgery, which involves stepwise removal of cancerous tissue layers with immediate microscopic examination to ensure complete tumor removal while preserving healthy tissue. The advantages of Mohs surgery include high cure rates, tissue preservation, real-time margin assessment, and precise mapping. However, the procedure is time-consuming, requires specialized training, and may have higher costs compared to alternative treatments. Perioperative management considerations, including preoperative assessment, anesthesia techniques, surgical procedure, wound care, and follow-up, are discussed to ensure optimal outcomes and patient satisfaction. Understanding the historical context, surgical technique, advantages, and disadvantages of Mohs surgery is crucial for healthcare professionals involved in the management of skin cancer patients.

Índice de sucesso de exodontias realizadas com extrator minimamente traumático / Exodonticssuccess index performed with a minimally traumatic extractor / Índice de éxito de exodonciarealizado com unextractor traumático mínimo

Introdução:Técnicas para extração dentária vêm sendo aperfeiçoadas objetivando um procedimento que diminua o esforço profissional, o tempo cirúrgicoeamenize as dores e os processos inflamatórios. Neste sentido os extratores minimamente traumáticos,com a exodontia vertical, propõem-se a preservar o osso alveolar e proporcionar uma recuperação mais rápida e confortável para o paciente. Objetivo:Avaliar a efetividade do kit para extração minimamente traumática da Maximus® (Contagem, Minas Gerais, Brasil) na exodontia de raízes residuais de incisivos, caninos e pré-molares unirradiculares. Metodologia:Trata-se de um ensaio clínico, prospectivo e analítico. Os pacientes foram operados utilizando o dispositivo, sendo avaliados os dados demográficos, tempo cirúrgico, dor e conforto após a cirurgia, bem como o grau de satisfação profissional com o uso do dispositivo. Para verificar diferenças significativas foi utilizado o teste de Mann-Whitney e a busca de associações foi realizada com o Exato de Fisher. Para todos os testes foi estabelecida uma significância com p<0,05.Resultados:Quarenta elementos foram removidos, o tempo cirúrgico foi em média 16,28 minutos, níveis de dor e conforto imediatamente após a cirurgia se mantiveram baixos (p<0,0001), e o grau de satisfação profissional se manteve alto (p<0,0001). A taxa de sucesso do dispositivo foi de 93,3% para os elementosincisivos e 20% para os elementos caninos e pré-molares (p<0,0001).Conclusões:A eficácia do extrator é determinada pelo tamanho da superfície radicular cobertas com fibras periodontais e a localização do dente. No entanto pode ser bem indicada no planejamento de reabilitações implantosuportadas em região anterior de maxila e mandíbula (AU).

Introduction:Techniques for tooth extraction have been improvedaiming at a procedure that reduces professional effort, surgical time, pain and inflammatory processes. In this sense, minimally traumatic extractors with vertical extraction, propose to preserve the alveolar bone and provide a faster and more comfortablerecovery for the patient. Objective:To evaluate the effectiveness of the Maximus® Minimally Traumatic Extraction Kit (Contagem, Minas Gerais, Brasil) in the extraction of uniradicular residual roots from incisors, canines and premolars.Methodology:Thisis a clinical, prospective and analytical trial. Patients were operated on using the device, and demographic data, surgical time, pain and comfort after surgery were evaluated, as well as the degree of professional satisfaction with the use of the device.To verify significant differences, the Mann-Whitney test was used and the search for associations was performed with Fisher's exact test. For all tests, significance was set at p<0.05.Results:Forty elements were removed, surgical time averaged 16.28 minutes, levels of pain and comfort immediately after surgery remained low (p<0.0001), and the degree of job satisfaction remained high (p<0.0001). The success rate of the device was 93.3% for the incisor elements and 20% for the canine and premolar elements (p<0.0001).Conclusions:The effectiveness of the extractor is determined by the size of the root surface covered with periodontal fibers and the location of the tooth. However, can be well indicated in planning implanted rehabilitation in the anterior region of the maxilla and mandible (AU).

Introducción: Se han mejorado las técnicas de extracciónde dientescon el objetivo de un procedimiento que reduzca el esfuerzo profesional, el tiempo quirúrgico, el dolor y los procesos inflamatorios. En este sentido, los extractores mínimamente traumáticos con extracción vertical tienen como objetivo preservar el hueso alveolar y proporcionar una recuperación más rápida y cómoda para el paciente. Objetivo: Evaluar la efectividad del Kit de Extracción Mínimamente Traumática Maximus® (Contagem, Minas Gerais, Brasil) en la extracción de raíces residuales de incisivos, caninos y premolares uniradiculares. Metodología: Es un ensayo clínico, prospectivo y analítico. Los pacientes fueron intervenidos con el dispositivo y se evaluaron datos demográficos, tiempo quirúrgico, dolor y comodidad después de la cirugía, así como el grado de satisfacción laboral con el uso del dispositivo. Para verificar diferencias significativas se utilizó la prueba de Mann-Whitney y la búsqueda de asociaciones se realizó mediante la prueba exacta de Fisher. Para todas las pruebas, la significancia se estableció en p <0,05. Resultados:Se retiraron cuarenta ítems, el tiempo quirúrgico promedió 16,28 minutos, los niveles de dolor y comodidad inmediatamente después de la cirugía permanecieron bajos (p<0,0001) y el grado de satisfacción laboral se mantuvo alto (p<0,0001). La tasa de éxito del dispositivo fue del 93,3% para los elementos incisivos y del 20% para los elementos caninos y premolares (p<0,0001).Conclusiones: La efectividad del extractor está determinada por el tamaño de la superficie radicular cubierta por fibras periodontales y la ubicación del diente. Sin embargo, puede resultar muy adecuado para planificar la rehabilitación con implantes en la región anterior del maxilar y la mandíbula (AU).

An optimized BRCA1/2 next-generation sequencing for different clinical sample types / 부인종양

Estabelecimento de protocolo para armazenamento de pele autógena refrigerada / Establishment of a protocol for storage of refrigerated autologous skin

Introdução: Enxertos de pele autólogos são utilizados em tratamento de pacientes queimados. Esses enxertos podem ser armazenados e preservados, desde que o processo de armazenamento seja realizado com rígido controle de qualidade, para garantir a redução dos riscos de infecção. Métodos: Foi realizado um estudo de coorte retrospectivo na Unidade de Queimados do Hospital das Clínicas de São Paulo no período de fevereiro de 2015 a julho de 2016, em que foi estabelecido um protocolo para armazenamento de pele refrigerada com controle de coleta, preservação, embalagem e registro de todos os processos. Para garantia de qualidade, foram coletadas biópsias dos enxertos para microbiologia pré e pós-armazenamento e realizado um estudo transversal de prevalência de contaminação pré e pós-estocagem. Resultados: Os pontos críticos encontrados foram inadequação de embalagem, ausência de registros de processos, falta de coleta de biópsias para microbiologia e falhas no descarte. A maior parte das amostras estava contaminada tanto pré como pós-estocagem (84,2%). Apenas dois pacientes apresentaram microbiologia estéril no pré e contaminada no pós, porém foram encontrados germes da pele do tipo gram+. Conclusão: Foi estabelecido um método promissor de armazenamento de pele refrigerada que necessita alguns pequenos ajustes para adequação ao controle de qualidade.

Introduction: Autologous skin grafts are used for treatment of burn patients. These grafts can be stored and preserved, as long as the storage process is performed with strict quality control to reduce the risk of infection. Methods: A retrospective cohort study was conducted in the Burn Unit of the Hospital das Clínicas de São Paulo from February 2015 to July 2016. During this period, a protocol was established to store refrigerated skin, with control of collection, preservation, and packaging, and recording of all processes. To ensure quality, graft biopsies were collected for pre- and poststorage microbiology testing and a cross-sectional study for contamination was performed. Results: Critical deficiencies included inadequate packaging, lack of processing records, lack of biopsies for microbiology testing, and failure to discard specimens. Most of the samples were contaminated before and after storage (84.2%). Only two samples were sterile before storage but became contaminated after storage, with growth of Gram-positive skin bacteria. Conclusion: A promising method for the storage of refrigerated skin was established, but requires minor adjustments in quality control.

Conceptos generales sobre técnicas de preservación de la fertilidad en la paciente oncológica / General Concepts about Techniques for Fertility Preservation in Cancer Patients

Actualmente, la necesidad de iniciar terapias antineoplásicas, no debe suponer la renuncia a la maternidad por parte de una paciente, que aún no haya completado sus deseos genésicos. Durante los últimos años, los avances socioeconómicos en los países desarrollados, han producido un retraso en la edad en las que las mujeres inician la búsqueda de su primera gestación. El objetivo del trabajo es mostrar una revisión pormenorizada de la literatura científica referente a la quimioprofilaxis con análogos de la GnRH, criopreservación de tejido ovárico y técnicas de estimulación ovárica para criopreservación de ovocitos y/o embriones, en pacientes con patología oncológica, sin deseos genésicos cumplidos. Se ha realizado una revisión bibliográfica de la literatura publicada en las bases de datos de PubMed, MedLine, Embase, BioMed Central y SciELO. Gracias a la mejoría de los tratamientos oncológicos, a los programas de detección precoz y a la aparición de nuevos fármacos y pautas terapéuticas, se ha incrementado la supervivencia de las pacientes con patologías oncológicas. Todo ello ha permitido el desarrollo de terapias genésicas óptimas, para este grupo de mujeres. La valoración inicial de estas pacientes debe incluir el grado de afectación de la función ovárica que les ocasionará el tratamiento y su repercusión en la reserva ovárica. La reserva ovárica es la cantidad de ovocitos que tiene la mujer en el momento del diagnóstico, ésta disminuye exponencialmente con la edad, por lo que es un factor muy importante a tener en cuenta.

Currently, the need to initiate antineoplastic therapies should not mean giving up on motherhood by a patient who has not yet fulfilled her desire to become a mother. In recent years, socioeconomic advances in developed countries have led to a delay in the age at which women begin their search for their first pregnancy. The objective of this paper is to show a detailed review of the scientific literature regarding chemoprophylaxis with GnRH analogues, cryopreservation of ovarian tissue and ovarian stimulation techniques for cryopreservation of oocytes and / or embryos, in patients with oncological pathology, who has not fulfilled their reproductive desires. A literature review was carried out in PubMed, MedLine, Embase, BioMed Central and SciELO databases. Thanks to the improvement of oncological treatments, early detection programs and the appearance of new drugs and therapeutic guidelines, the survival of patients with oncological pathologies has increased. All this has allowed the development of optimal gene therapy for this group of women. The initial assessment of these patients should include the degree of ovarian function impairment that will cause the treatment and its impact on the ovarian reserve. The ovarian reserve is the number of oocytes that the woman has at the time of diagnosis, this decreases exponentially with age, which is a very important factor to take into account.

Preservación de biopsias obtenidas por colonoscopia en pacientes con cáncer colorrectal para análisis moleculares / Preservation of obtained biopsies by colonoscopy in patients with colorectal cancer for molecular analysis

Introducción: los estudios relacionados con el análisis del ADN han marcado una pauta para los avances en las ciencias básicas y uno de los requisitos para la obtención de buenos resultados es la calidad del material genético extraído, en conjunto con el método empleado para la preservación de las muestras. Objetivo: comparar tres métodos de preservación de biopsias obtenidas por colonoscopia en pacientes con cáncer colorrectal con fines de uso en estudios de biología molecular. Materiales y métodos: se tomaron biopsias por colonoscopia a nueve pacientes con diagnóstico clínico de cáncer de colon, las cuales se preservaron en solución salina y dos solventes estabilizadores de ácidos nucleicos, RNAlater® y LifeGuard™ Soil Preservation Solution; se realizó extracción del ADN total a las nueve muestras y se verificó la concentración y la calidad del ADN extraído. Resultados: la extracción del ADN a las 24 horas, ocho días, quince días, un mes, seis meses, uno y dos años después de la toma de la muestra, mostró que el ADN de las biopsias preservadas en solución salina se presentaba con baja concentración y degradado a los ocho días, mientras que el preservado en soluciones comerciales estabilizadoras presentó una buena calidad y alta concentración. Por otro lado, la calidad del ADN fue verificada mediante la amplificación por reacción en cadena de la polimerasa (PCR) de un fragmento de ADN asociado al gen APC. Conclusiones: las soluciones LifeGuard™ y RNAlater® pueden ser usadas durante el transporte y conservación de tejidos humanos, y pueden ser recomendados para aquellos laboratorios que deseen preservar muestras con métodos diferentes al embebido de muestras en parafina o que no cuentan con métodos de conservación altamente eficientes como la criogénesis. (AU)

Introduction: Studies related to DNA analysis have set a standard for development in the basic sciences, and one of the requirements for obtaining good results is the quality of the genetic material extracted, together with the method used for the samples preservation. Objective: To compare three methods of preserving biopsies obtained by colonoscopy in patients with colorectal cancer, for purposes of use in molecular biology studies. Materials and methods: Biopsies by colonoscopy from nine patients with clinical diagnosis of colon cancer were taken and preserved in saline solution and two nucleic acids stabilizing solvents, RNAlater® y LifeGuard™ Soil Preservation Solution. The extraction of total DNA was carried out to the nine samples and the concentration and quality of the extracted DNA was verified. Results: DNA extraction at 24 hours, eight days, 15 days, one month, six months, one year and two years after sampling showed that the DNA from biopsies preserved in saline solution had low concentration and degraded at eight days. DNA from samples preserved in commercial stabilizing solutions had high concentration and good quality. This last one was verified by amplification of a DNA fragment associated to the APC gene by polymerase chain reaction. Conclusions: The LifeGuard™ solution as well as the RNAlater® can be used in transport and conservation of human tissues, and may be recommended for laboratories wishing to preserve specimens with different methods to paraffin-embedded specimen or that do not have highly efficient conservation methods such as cryogenics. (AU)

Protocolo para a captação, transporte e armazenamento de tecido osteocondral humano / Evaluation of the protocol for harvest, transport and storage of human osteochondral tissue

O tratamento das lesões osteocondrais representa um desafio ao cirurgião ortopédico. Diversas técnicas biológicas visam a restauração do tecido lesionado, estando limitadas a formação de fibrocartilagem ou a dimensão da área a ser restaurada. O transplante alógeno osteocondral representa o único método biológico capaz de fornecer cartilagem hialina para o tratamento das lesões maiores que 2cm². A viabilidade celular do tecido no momento da cirurgia representa um fator indispensável para que haja sucesso no tratamento. A ausência de um protocolo definido pelos órgãos reguladores de saúde nos levou a necessidade de elaborar uma metodologia própria para captação, transporte e armazenamento de tecido osteocondral humano. Foram analisados fragmentos osteocondrais com dimensão de 2cm³ de cinco doadores cadáveres com idade entre 15 e 45 anos. As amostras foram armazenadas em meio de preservação celular contendo: albumina humana, Iscove's e vancomicina preservados à temperatura de 4°C. A concentração de proteoglicanos no meio extracelular foi quantificada pelo uso de Safranina-O, enquanto a análise estrutural do tecido foi avaliada através de estudo histológico com lâminas coradas em hematoxilinaeosina. As imagens obtidas foram analisadas segundo os escore histológicos de Mankin e o escore proposto pela OsteoArthritis Research Society International. As amostras foram analisadas com 0, 15, 30 e 45 dias de preservação. Os fragmentos osteocondrais estudados apresentaram diminuição progressiva na concentração de proteoglicanos com o aumento do tempo de preservação, soretudo na camada de superficial da cartilagem. Após 30 dias de preservação foram identificadas alterações estruturais com descontinuidade da camada superficial da cartilagem. As demais camadas do tecido osteocondral mantiveram suas estruturas preservadas após 45 dias de preservação. Segundo os resultados obtidos pelo escore de Mankin, houve diferença com significância estatística entre 15 e 30 dias de preservação do tecido. Os resultados encontrados definiram o protocolo de preservação transporte e armazenamento de tecido osteocondral humano a ser aplicado pelo Banco de Tecidosmultiesquelético lotado no Instituto Nacional de Ortopedia e Traumatologia.

The treatment of osteochondral lesions remains a challenge to the orthopedic surgeon. Various biological techniques aim at the restoration of the injured tissue, being limited the formation of fibrocartilage or the size of the area to be restored. The allogeneic osteochondral transplantation represents the only biological method capable of providing hyaline cartilage after lesions larger than 2cm². The cellular viability of the tissue at the time of surgery is an essential factor for successful treatment. The absence of a protocol defined by the health regulatory agencies has led us to the need to develop a proper methodology for harvest, transporting and storing human osteochondral tissue. Osteochondral fragments measuring 2cm³ of five cadaveric donors aged between 15 and 45 years were analyzed. The samples were stored in cell preservation medium containing: human albumin, Iscove's and vancomycin maintained at 4 ° C. Immunohistochemical study with antibodies to the Caspase 3 protein evaluated the presence of cell death by apoptosis. The concentration of proteoglycans in the extracellular matrix was quantified by the use of Safranin-O, while tissue structural analysis was assessed by histological study with hematoxylin-eosin stained slides. The images obtained were analyzed according to the histological scores of Mankin, OsteoArthritis Research Society International and the score proposed by the authors. The samples were analyzed with 0, 15, 30 and 45 days of preservation. The samples studied presented a progressive decrease in the concentration of proteoglycans with the increase of preservation time. After 30 days of preservation, structural changes were identified in the superficial layer of the cartilage. The remaining layers of the osteochondral tissue maintained their structures preserved after 45 days of preservation. According to the results obtained by the Mankin score, there were differences with statistical significance between 15 and 30 days of preservation. The results found defined the protocol for the preservation of the transport and storage of human osteochondral tissue to be applied by the Bank of Tissues, located in the National Institute of Orthopedics and Traumatology

Nuevo método de inclusión en resina poliéster P-4 para cortes anatómicos / New method for P-4 polyester resin inclusion for anatomical slices

La resina poliéster (P-4) se ha utilizado en varias técnicas anatómicas, gracias a su bajo costo, fácil manejo y obtención. Entre ellas la inclusión, que se realiza en capas dejando muestras relativamente opacas y de un grosor que no permite ver claramente las estructuras. También encontramos la plastinación de cortes anatómicos, técnica más sofisticada, compleja y de un alto costo económico. El objetivo fue idear un método de inclusión para cortes anatómicos, que mejore la calidad de la técnica clásica. Se utilizaron cortes anatómicos de segmento distal de miembro inferior humano y de un grosor promedio de 0,5 cm fijados con acetona y cortes de encéfalo de vacuno de igual grosor, fijados con formalina acuosa al 10%. La totalidad de las muestras son deshidratadas en acetona a -17 °C. Posteriormente se impregnan e incluyen en resina poliéster (P-4) a temperatura ambiente. Se obtuvieron muestras con una buena transparencia y solidez que permite ver en detalle muchas estructuras, similar a la obtenida con la técnica de plastinación en resina. Mediante un método relativamente fácil de realizar es posible obtener piezas anatómicas de muy buena calidad.

The polyester resin (P-4) has been used in several anatomical techniques, due to its low cost, easy handling and elicit. Among them, the inclusion, which is performed on samples leaving relatively opaque layers and an important thickness which does not allow a clear vision of the structures. There is also the plastination of anatomical slices with this polymer which although, technically sophisticated and complex has a high economic cost. The objective is to develop a method for anatomical section inclusion, improving the quality of classic technique. Distal segment of human lower limb anatomical slices were used with an average thickness of 0.5 cm fixed with acetone and bovine brain slices equal thickness, fixed with 10% aqueous formalin. All the samples are dried in acetone at -17 °C. Subsequently, they are impregnated and included on polyester resin (P-4) at room temperature. Samples were obtained with good transparency and solidity that allows observation of details of many structures, similar to that obtained with the resin plastination technique. Using this relatively easy method we can get very good quality anatomical samples.

Causas do descarte de córneas captadas pelo banco de tecidos oculares do Rio Grande do Norte / Causas de descarte de córneas procesados por el banco de tejidos oculares de Rio Grande do Norte / Causes for the discards of corneas collected by the ocular tissues bank from Rio Grande do Norte

Objective: To identify the causes for the discards of corneas in the ocular tissue bank from Rio Grande do Norte. Method: This was an exploratory descriptive study, with retrospective data and quantitative approach, conducted in the ocular tissue bank from Rio Grande do Norte. The information was collected by means of an instrument containing: number of collected, transplanted, and discarded corneas; causes for the discards. Results: Out of the 548 collected corneas, 78.1% were preserved and 21.9% discarded.The prevalent causes for the discards were: stromal infiltrate, positive serology, and expired validity. Conclusion: Health professionals should be prepared to decrease the loss of corneas for avoidable reasons...

Objetivo: Identificar as causas dos descartes de córneas no banco de tecidos oculares do Rio Grande do Norte. Método: estudo exploratório descritivo, com dados retrospectivos e de abordagem quantitativa,realizado no banco de tecidos oculares do Rio Grande do Norte. As informações foram coletadas por meio de instrumento contendo: número de córneas preservadas, transplantadas e descartadas; causas dos descartes. Resultados: das 548 córneas captadas, 78,1% foram preservadas e 21,9% descartadas. As causas prevalentes dos descartes foram: infiltrado estromal, sorologia positiva e validade inspirada. Conclusão: os profissionais da saúde devem diminuir a perda de córneas por motivos evitáveis...

Objetivo: Identificar las causas de los descartes de las córneas de un banco de tejidos oculares de RioGrande do Norte. Método: Estudio exploratorio descriptivo con los datos retrospectivos y enfoque cuantitativo llevado a cabo los tejidos oculares de stock de Rio Grande do Norte. La información se recogió mediante un instrumento que contiene: número de córneas trasplantadas conservadas y descartadas;causas de los descartes. Resultados: De 548 trasplantes de córnea, el 78,1% y el 21,9% fueron preservados descartado. Las causas más frecuentes de los descartes fueron: infiltrado estromal, serología positiva y la validez de inspiración. Conclusión: Los profesionales de salud deben reducir la pérdida de córneas por razones evitables...

Use of cryopreserved cadaveric arterial allograft as a vascular conduit for peripheral arterial graft infection

Major peripheral arterial graft infection is a potentially devastating complication of vascular surgery, associated with significant mortality and high amputation rates. Autologous saphenous veins are considered optimal arterial conduits for lower extremity revascularization in infected fields, but they are often unavailable or unsuitable in these patients. This study describes two patients with major peripheral graft infection, but without available autologous veins, who underwent graft excision and cryopreserved cadaveric arterial allograft reconstruction. Although long-term graft durability is unclear because of gradual deterioration and degeneration, these findings suggest that cadaveric allografts may be good options for patients with major peripheral graft infection.

Características das doações de córnea no estado do Piauí / Characteristics of corneal donations in state of Piauí / Characteristics of corneal donations in state of Piauí

Objetivo: Traçar o perfil das doações de córnea obtidas pelo Banco de Tecidos Oculares do Hospital Getúlio Vargas (BTOC-HGV), avaliar a qualidade do tecido captado e enumerar as indicações do transplante de córnea. Métodos: Estudo retrospectivo de ficha de doadores cadastrados no BTOC-HGV no período de 2008 a 2011. Foram coletados dados de idade, gênero e causa do óbito do doador, tempo de enucleação, tempo de preservação, qualidade da córnea doada, causas de descartes e indicações de transplante de córnea no BTOC. Resultados: Foram analisadas 311 fichas de doadores de córnea do BTOC-HGV. O número de doações aumentou de 9 em 2008 para 80 em 2009, 109 em 2010 e 113 em 2011. A maioria dos doadores era do gênero masculino. A média da idade dos doadores foi de 43,00 ± 16,01 anos. A maioria das córneas transplantadas (78,6%) foi óptica. Foram descartadas 7,16% das córneas ópticas, sendo a maioria por nova classificação em tectônica e detecção de sorologia positiva. O tempo de enucleação foi de 3,77 ± 2,18 horas e o de preservação foi de 6,86 ± 6,10 horas. A causa mortis mais frequente foram causas externas, seguida por doenças do aparelho circulatório, respiratório, digestivo e geniturinário. A principal indicação de transplante foi ceratopatia bolhosa, seguida por ceratocone e perfuração corneana. Conclusão: Este estudo demonstrou a existência de fatores associados à qualidade das córneas captadas pelo BTOC como idade, tempo entre a enucleação e a preservação e causa do óbito. A principal indicação de transplante foi ceratopatia bolhosa. .

Objective: To describe the profile of corneal donations obtained by Ocular Tissue Bank, Hospital Getúlio Vargas (BTOC-HGV), assess the quality of the captured tissue and list the indications for corneal transplant. Methods: Retrospective study of registered donors in BTOC-HGV in the period 2008-2011. Collected data were: age, gender and cause of death of the donor, enucleation time, preservation time, quality of the donated cornea, causes of discards and indications for penetrating keratoplasty in BTOC. Results: Were analyzed 311 records of donor cornea BTOC-HGV. The number of donations increased from 9 in 2008 to 80 in 2009, 109 in 2010 and 113 in 2011. Most donors were male. The average age of the donors was 43.00 ± 16.01 years. The majority of transplanted corneas (78.6%) was optics. Were discarded 7.16% of corneal optics, mostly by new tectonic classification and detection of positive serology. The time of enucleation was 3.77 ± 2.18 hours and preservation was 6.86 ± 6.10 hours. The most frequent causes of death were external causes, followed by diseases of the circulatory, respiratory, digestive and genitourinary systems. The main indication for transplantation was bullous keratopathy, followed by keratoconus and corneal perforation. Conclusion: This study demonstrated the existence of factors associated with quality of corneas by BTOC as age, time between enucleation and preservation and cause of death. The main indication for transplantation was bullous keratopathy. .

Objective: To describe the profile of corneal donations obtained by Ocular Tissue Bank, Hospital Getúlio Vargas (BTOC-HGV), assess the quality of the captured tissue and list the indications for corneal transplant. Methods: Retrospective study of registered donors in BTOC-HGV in the period 2008-2011. Collected data were: age, gender and cause of death of the donor, enucleation time, preservation time, quality of the donated cornea, causes of discards and indications for penetrating keratoplasty in BTOC. Results: Were analyzed 311 records of donor cornea BTOC-HGV. The number of donations increased from 9 in 2008 to 80 in 2009, 109 in 2010 and 113 in 2011. Most donors were male. The average age of the donors was 43.00 ± 16.01 years. The majority of transplanted corneas (78.6%) was optics. Were discarded 7.16% of corneal optics, mostly by new tectonic classification and detection of positive serology. The time of enucleation was 3.77 ± 2.18 hours and preservation was 6.86 ± 6.10 hours. The most frequent causes of death were external causes, followed by diseases of the circulatory, respiratory, digestive and genitourinary systems. The main indication for transplantation was bullous keratopathy, followed by keratoconus and corneal perforation. Conclusion: This study demonstrated the existence of factors associated with quality of corneas by BTOC as age, time between enucleation and preservation and cause of death. The main indication for transplantation was bullous keratopathy. .

Experiencia en plastinación con resina poliéster P-4 para cortes anatómicos / Plastination experience in polyester resin (P-4)

El Laboratorio de Plastinación de la Universidad de los Andes ha trabajado hasta hoy, exclusivamente la técnica en Silicona para preservar el material cadavérico mediante este tipo de conservación. Sin embargo, este método tiene algunas desventajas en cortes anatómicos finos. Se desarrolló un método de plastinación en resina poliéster, en base a la técnica ya protocolizada, adaptándolas a nuestro laboratorio, con el fin de obtener muestras con mayor resistencia y calidad, sin alterar las estructuras. Se utilizaron cortes anatómicos de segmentos distales de miembro inferior humano fijados en acetona a - 17 C y otros de encéfalo de vacuno fijados con formalina acuosa al 10 por ciento a 4 C, de un grosor que varía entre 0,4 y 0,8 cm. Se procedió a deshidratar las muestras en acetona a -17 C para luego seleccionar piezas que fueron impregnadas y plastinadas con resina P-4 sin catalizador y otras con catalizador en proporciones 2:1. El proceso de polimerización fue común para ambas con luz ultravioleta. Se obtienen muestras sólidas y rígidas, con cierta transparencia, especialmente en los cortes más finos. No se observaron alteraciones de forma y tamaño. Los mejores resultados en cuanto a transparencia, se obtuvieron con la Resina sin catalizador. Se logró estandarizar una técnica con Resina poliéster P-4, obteniendo cortes anatómicos de buena calidad y con una resistencia mucho mayor que la obtenida para este tipo de muestras en la plastinación con Silicona...

The plastination laboratory at Universidad de los Andes has always worked exclusively silicone as a technique to preserve cadaveric material. However, this method has disadvantages in thin anatomical pieces. A plastination method has been developed using polyester resin P-4, using an already protocolized technique and adapting it to our laboratory to obtain better quality and resistant pieces, without altering the anatomical structures. Anatomical pieces of human lower extremities were used, which were fixated in acetone at -17 C and bovine encephalous pieces, fixated with 10 percent aquoeus formalyn at 4 C, with a variable width of 0.4 to 0.8 cm. The pieces were then dehydrated in acetone at -17 C; among these pieces, some were selected, impregnated and plastinated with P-4 resin, using in some of them a catalyst in a ratio of 2:1. The polymerization process was the same for all pieces with ultraviolet light. Solid and rigid pieces were obtained, with a partial transparency, especially on the thinnest cuts. No alterations in size and shape were observed. The best results in transparency were obtained with the technique without the use of catalyst. An adapted standardized protocol for polyester resin P-4 technique was achieved, obtaining good quality anatomical pieces with a higher resistance compared to pieces plastinated using the silicone technique...

Características dos doadores de córneas do Banco de Olhos de Cascavel: impacto do exame anti-HBc para hepatite B / Characteristics of corneal donors in the Cascavel Eye Bank: impact of the anti-HBc test for hepatitis B

OBJETIVOS: Traçar o perfil dos doadores de tecidos oculares humanos na região de atuação do Banco de Olhos de Cascavel - Paraná (PR) e verificar o impacto da positividade da sorologia positiva para hepatite B (anti-HBc) no descartes dos tecidos oculares. MÉTODOS: Estudo do tipo transversal de dados do Banco de Olhos de Cascavel do período de março de 2006 a abril de 2007. Informações como idade, sexo, causa de óbito, tempo entre óbito e enucleação e resultados sorológicos de doadores foram coletadas. Dados foram agrupados para caracterização da população de estudo e distribuídos segundo positividade da sorologia para hepatite B (anti-HBc). RESULTADOS: A média de idade dos doadores foi de 54,2 ± 20,6 anos e a maioria era do sexo masculino (64,7 por cento). Tempo médio entre óbito e enucleação foi de 3,8 ± 2,4 horas, a principal causa de óbito foram doenças cardiovasculares (34,5 por cento). Positividade para os marcadores anti-HBc e HBsAg foi de 47,4 por cento e 1,5 por cento, respectivamente, sendo os exames sorológicos para hepatite B a maior causa de descarte dos tecidos oculares doados. Doadores com sorologia positiva para anti-HBc apresentaram maior média de idade comparados com doadores com sorologia negativa (p<0,001). Somente 0,8 por cento das doações apresentaram sorologia positiva para hepatite C. Não houve doações com sorologia para HIV positiva no período do estudo. CONCLUSÕES: Implementação do Banco de Olhos de Cascavel aumentou significativamente a disponibilização de córneas para transplante no Estado do Paraná. A principal contraindicação para disponibilização de tecidos oculares foi a positividade na sorologia para anti-HBc. Estudos de biosegurança devem ser realizados a fim de investigar o potencial de transmissão da hepatite B de doadores positivos para esse marcador. A inclusão desses doadores como doações efetivas acarretaria maior disponibilização de tecidos para transplantes.

PURPOSE: To describe the characteristics of human ocular tissue donors from the Cascavel Eye Bank in the State of Paraná (PR) and investigate the impact of hepatitis B serologic test positivity on discarded ocular tissues. METHODS: Cross-sectional study of data collected between March 2006 and April 2007 at the Cascavel Eye Bank. Information such as age, gender, cause of death, time between death and enucleation, and serologic tests results was collected. Data were used to characterize the study population and stratified according to hepatitis B (anti-HBc) status. RESULTS: Mean age of donors was 54.2 ± 20.6 years and a greater proportion of men (64,7 percent) was observed. Mean time between death and enucleation was 3.8 ± 2.4 hours, cardiovascular diseases were the main cause of death (34.5 percent). Positivity of anti-HBc and HBsAg tests was 47.4 percent and 1.5 percent, respectively. Hepatitis B was the main reason for discarding corneas. Donors whose serologic tests were positive for anti-HBc had higher mean age compared to those with negative tests (p<0.001). Only 0.8 percent of all donations tested positive for hepatitis C and donors with positive HIV test were not seen in our study. CONCLUSIONS: Activities of the Cascavel Eye Bank significantly increased the availability of corneas for transplant in the State of Paraná. The main contraindication for availability of ocular tissues was positivity of serologic tests for anti-HBc. Biosafety studies should be performed in order to investigate the potential transmission of hepatitis B virus for donors with positive tests for anti-HBc. The inclusion of these donors would increase the number of corneas available for transplant.

Comparison of the DNA Preservation in Neutral-Buffered Formalin Fixed Paraffin-Embedded Tissue and in Non-Buffered Formalin Fixed Paraffin-Embedded Tissue

BACKGROUND: The preservation of optimized DNA and its extraction from formalin-fixed, paraffin-embedded (FFPE) tissues are important issues. There has been some doubt over whether 10% neutral-buffered formalin is an ideal fixation solution for DNA preservation over non-buffered formalin, as conventionally recommended. In this study, the correlation between the efficiency of DNA extraction from FFPE tissues and buffered formalin was evaluated. METHODS: Several tissues with same conditions except fixatives were fixed in four different formalin solution groups and were routinely processed as paraffin-embedding protocols. DNAs were extracted from four different FFPE tissues that were stored for over 3 months and over 9 months. The quantity and quality of the DNAs were assessed with a NanoDrop ND-1000 spectrophotometer, and the polymerase chain reaction (PCR) amplification and degradation were analyzed via microchip electrophoresis. KRAS mutation analysis and microsatellite instability (BAT25) PCR were performed with each sample. RESULTS: The results showed no remarkable difference in the four groups. CONCLUSIONS: The study findings demonstrate that DNA preservation is fairly unaffected by a neutral buffer where there is short formalin manufacture period and an adequate formalin fixation time before embedding in paraffin.

Effect of loading environment on the human osteochondral allograft preserved by tissue culture / 中华创伤杂志

Objective To study the effect of the loading environment on the chondrocytes and the collagen ultrastructure at articular cartilage of the osteochondral allograft graft preserved by tissue culture method so as to improve the preservation effect. Methods A kind of shaking table was used as a dynamic loading device for storage of the osteochondral allograft to simulate the mechanical environment of normal articular cartilage in vivo. The device worked in an incubator at 3711 and the frequency was 1Hz. Western blotting was used for analysis of β-actin at the articular cartilage of the graft for investigating the effects of stress condition on the chondrocyte function. Transmission electron microscopy was used to observe the collagen ultrastructure at the articular cartilage of the graft two weeks after storage and compared with the control group. Results The dynamic loading condition could significantly increase the expression of β-actin and keep the ultrastructure of the collagen fibril in articular cartilage compared with the static condition. Conclusions The loading condition benefits the chondrocyte function and collagen ultrastructure of the articular cartilage in human osteochondral allograft. The dynamic mechanical loading may provide a better method in preservation of the human osteochondral allograft.

Estudio comparativo de dos técnicas de plastinación / Two different plastination techniques comparative study

La técnica de plastinación es utilizada ampliamente para la mejor preservación de las piezas cadavéricas utilizadas en docencia. Nuestra unidad la está utilizando desde el año 2002, pero hemos notado que la calidad de las muestras no es la óptima, produciéndose gran retracción de éstas. Por tal motivo, se diseñó una trabajo que compara la técnica de plastinación de nuestra unidad, con otro proceso, el cual se basa en el protocolo utilizado por la Universidad de Murcia. Utilizamos 24 muestras frescas de riñones y hemiencéfalos de vacuno, riñones y corazones de cerdo y miembro inferior humano, las cuales fueron fijadas con alcohol etílico, formalina y solución fijadora Universidad de los Andes. Fueron aplicadas dos diferentes técnicas de plastinación: una basada en el protocolo utilizado hasta ese momento en nuestro laboratorio de plastinación (técnica A) y otro proceso similar al publicado por la Universidad de Murcia (técnica B). Se evaluaron los porcentajes de variación de peso, ancho, largo, espesor y perímetro de las diferentes piezas plastinadas. También se procedió a fijar las muestras en diferentes soluciones (alcohol, formalina 10 por ciento y solucion fijadora Universidad de los Andes), pesar y medir sus dimensiones. El porcentaje de pérdida de peso promedio fue 63,1 por ciento para la técnica A y 37,9 por ciento para la técnica B (p<0,01); las mediciones de las disminuciones en el largo, ancho y grosor de las muestras también fueron mayores en las piezas sometidas a la técnica A, siendo todos los porcentajes de pérdidas estadísticamente significativos. Al comparar las muestras según los diferentes métodos de fijación con la técnica B no hubo diferencias estadísticamente significativas. La plastinación de muestras basadas en la técnica de Murcia obtiene muestras de mejor calidad y menos retracción. No influiría en el proceso de plastinación el método de fijación.

The plastination technique is used extensively for the improved preservation of cadavers used for teaching. Our unit has been using this technique since 2002, but we have noticed that sample quality is not optimal, given the large retraction of samples. Therefore, a project was designed that compares our department's plastination technique, with another process, based on the protocol used by Murcia University. We used 24 fresh bovine kidneys and brain samples, pigs' kidneys and hearts and human lower limbs; all of which were fixed with ethyl alcohol, formalin and De los Andes University's fixation solution. Two different plastination techniques were applied: one based on the protocol used in our laboratory (technique A) and another one similar to that published by Murcia University (technique B). We evaluated the body weight percentage, width, length, thickness and perimeter of the different plastinated pieces, as well as measuring the same variables in each fixation solution (alcohol, formalin fixative solution 10 percent and Universidad de los Andes). The average weight loss percentage was 63.1 percent for technique A and 37.9 percent for technique B (p <0.01). The decrease measurements in length, width and thickness were also higher under technique A, all percentages being statistically significant. When comparing the samples according to different fixation methods, there were no significant differences. Plastination based on Murcia University's technique obtained better quality samples with less shrinkage. The plastination process was not influenced by the fixation method.

Análise das córneas do Banco de Olhos da Santa Casa de São Paulo utilizadas em transplantes / Analysis of the transplanted corneas at Santa Casa de São Paulo Eye Bank

Objetivo: Traçar o perfil do doador de olhos e avaliar os fatores que influenciam na qualidade das córneas captadas pelo Banco de Olhos da Santa Casa de Misericórdia de São Paulo (SCMSP), no período de junho de 2000 a janeiro de 2004. Métodos: Foram estudados os dados dos prontuários de 151 pacientes submetidos a transplante penetrante de córnea, cadastrados no Banco de Olhos da SCMSP, no período entre junho de 2000 e janeiro de 2004, totalizando 180 operações. Resultados: A média da idade dos doadores foi de 55,57 ± 19,74 anos (variando de 9 meses a 97 anos de idade), sendo 54,44 por cento dos pacientes do sexo masculino. As causas mortis predominantes foram as cardiovasculares, com 50,55 por cento, seguidas das doenças consumptivas 17,77 por cento, traumáticas 14,44 por cento e outras, que somadas representam 17,22 por cento. Em relação à qualidade da córnea, não houve diferença estatística significativa em relação aos quatro grupos causas mortis p=0,527), e o tempo decorrido entre o óbito e a preservação (p=0,053). Houve diferença estatisticamente significativa entre a qualidade das córneas de doadores da faixa etária acima de 61 anos de idade em relação às outras faixas etárias (p= 0,037). Conclusão: As causas mortis mais frequentes foram as cardiovasculares, seguidas das causas pulmonares, outras e traumáticas. Observamos também que entre os fatores inerentes ao doador, a faixa etária acima de 60 anos, predispôs à pior qualidade anatômica das córneas, e fatores como a causa mortis e o tempo decorrido entre o óbito e a preservação, não influenciaram na qualidade das mesmas.

Purpose: To identify the donors' profile and evaluate the factors influencing the quality of the donated corneas in Santa Casa Eye Bank, from July, 2000 to January, 2004. Methods: Data from the charts of 151 patients submitted to penetrating keratoplasty, registered in Santa Casa Eye Bank, from July, 2000 to January 2004 resulted in 180 surgeries. Results: Mean age of the donors was 57.57 ± 19.74 years old (varied from 9 months to 97 years old) and 54.44 percent were male. The cause of death was predominantly cardiovascular (50.55 percent), followed by consumptive diseases (17.77 percent), trauma (14.44 percent) and others (17.22 percent). Considering the quality of the cornea, there was no statistically significant difference among the three main groups of cause of death (p=0.527) and time from death to corneal preservation (p=0.053). There was a statistically significant difference between quality of donated corneas above 61 years old and other ages (p=0.037). Conclusion: The most frequent cause of death was cardiovascular, followed by pulmonary diseases and trauma. We could also note that factors inherent to donor like age (>60 years old) predisposed to a worse anatomical quality of the cornea and factors like cause of death and time between death and preservation did not influence the quality of the cornea.

Experiment study on corneal moist chamber storage with aqueous removed and recipient serum tamponaded / 中国医师杂志

Objective To explore the new preservation method of cornea by evaluating the structure and function of rabbit's endothelia on the condition of whole-eye preservation with aqueous removed and recipient's serum tamponaded. Methods Forty New Zealand big white rabbits (80 eyes) were random divided into two groups, 40 eyes in group A (control group) and 40 eyes in group B (experimental group). The vitality of endothelial cell on the condition of tow corneal preservation methods which were the moist chamber preservation (group A) and the whole-eye preservation with aqueous removed and recipient's serum tamponaded (group B) was compared. At the 2nd day, the 5th day, the 7th day, the 10th day, the vitality of endothelial cell was appraised through ultra-microstructure by scanning electron microscope and the trpan biuealizarin red stain. The corneal thickness was measured, and corneal endothelial cells density was calculated, and cell size was observed by image analysis system. Result In group B, the corneas remained transparent for 7 days , and the rate of vitality was 90% for7 days and that was over 80% for 10 days. In group A, the corneas remained transparent for 2 days, and the endothelial cell losing and dying were found after 5 days. Cell vitality, cell density and cell size had no statistical difference between Group B for 7 days and group A for 2 days. (all P＞0. 05). In group A, at the 5th day, the 7th day ,the 10th day, the average corneal thickness were (0.64 ± 0.04) mm, (0. 79 ± 0. 03) mm , (1.06 ± 0. 03) mm. In the group B, at the 5th day, the 7th day ,the l0th day, the average corneal thickness were (0. 55 ±0.03)mm, (0.65 ±0. 02) mm , (0. 85 ± 0. 05) mm. The average corneal thickness had significant difference between group A and group B (all P ＜ 0. 05). Conclusion Recipient serum had the function with supporting the structure and function of rabbit's corneal endothelial cell, and it could prolong the storage time with the moist chamberstorage at the same time.

Comparison of Different Methods of Aortic Valve Conduit Xenograft Preservation in an Animal Experiment Model; Fresh Cryopreservation versus Acellularized Cryopreservation / 대한흉부외과학회지

BACKGROUND: The commercially used vascular xenografts have some problems such as calcification, fibrosis and tissue degeneration that are associated with inflammatory and immunologic reactions. We compared two methods of xenograft preservation (fresh cryopreservation versus acellularized cryopreservation) of goat aorta. MATERIAL AND METHOD: Aortic valved xenografts were harvested from adult pigs, and these were preserved using fresh cryopreservation (FC group, n=4) or acellularized crypreservation (AC group, n=4). These xenografts were implanted into adult goats. There were 2 short-term survivors (less than 100 days) and 2 long-term survivors in each group. These xenografts were explanted and they underwent microscopic examination. RESULT: The goats survived 31, 40, 107 and 411 days in the FC group and the other goats survived 5, 40, 363 and 636 days in the AC group. All the short-term survivors in each group expired because of rupture at the proximal anastomosis site. Marked neutrophil infiltration was observed in the FC group FC and lymphocytes were observed in the AC group. There were no differences in the occurrence of calcification, fibrosis and thrombosis among the groups. CONCLUSION: Some goats survived more than 100 days after the xenograft implantation irrespective of the methods of preservation. Because severe tissue degeneration developed in both groups, we think these methods are not appropriate for xenograft preservation of aorta. It was worth a preliminary trial for improving the preservation method or to modify the processing of xenografts.

Assessment of the use of cryopreserved x freeze-dried amniotic membrane (AM) for reconstruction of ocular surface in rabbit model / Avaliação do uso da membrana amniótica (MA) criopreservada x liofilizada na reconstrução da superfície ocular em coelhos

PURPOSE: To determine the efficacy of freeze-dried amniotic membrane (AM) for reconstruction of the ocular surface in rabbit eyes. METHODS: The sterilized, freeze-dried amniotic membrane (lyophilized or FD-AM) is a preservative method that uses the drying by freezing process to maintain the AM well preserved for a long time even at room temperature. This paper is an experimental animal interventional study. One eye of each of 15 male New Zealand rabbits (1.5 - 3.0 kg) had the central cornea marked with a 6.0 mm trephine. The marked area was deepithelialized with a No.15 blade. The denuded corneal surface was covered as follows: Group 1: cryopreserved AM (n=6); Group 2: freeze-dried AM (n=6); and Group 3: not covered (control group, n=3). The AM in group 1 and 2 and the periphery of the denuded area in group 3 were secured with continuous 10-0 nylon sutures. The clinical evaluation was made by a blinded observer and graded on a four-point scale (1= minimal, 4= marked) for conjunctival and ciliary hyperemia, eyelid edema, corneal neovascularization, corneal opacity and reepithelialization on postoperative (PO) days 1, 7 and 30 . After PO day 30, the rabbits were euthanized and their corneas were sent for histopathological and ultrastructural analysis to evaluate tissue inflammation, reepithelialization, and basement membrane integrity. RESULTS: Two eyes in group 2 had a corneal infection and were excluded from the analysis. No statistically significant differences among the three groups were found (p>0.05) regarding the clinical evaluation on 1st, 7th and 30th PO days. On transmission electron microscopy, the basement membrane in lyophilized and control groups was more continuous and homogeneous than in the glycerol group. CONCLUSIONS: The freeze-drying method seems to be a good option to preserve human amniotic membrane to be used in ocular surface reconstruction. This preservative method reduces the preservation costs and may enhance...

OBJETIVO: Avaliar a eficácia da liofilização da membrana amniótica (MA) para a reconstrução da superfície ocular em coelhos. MÉTODOS: A liofilização é processo de preservação que mantém a MA estável durante longo tempo mesmo em temperatura ambiente. A córnea de um olho de cada coelho macho da raça Nova Zelândia foi marcada e desepitelizada. Essa área desepitelizada foi coberta com: Grupo 1: MA criopreservada (n=6); Grupo 2: MA liofilizada (n=6) e Grupo 3: Não coberta (n=3). A MA nos grupos 1 e 2 e a periferia da córnea no grupo 3 foram suturadas com nylon 10-0. A avaliação clínica foi realizada por um observador cego em relação à hiperemia, neovascularização e edema de córnea e reepitelização nos dia 1, 7 e 30 pós-operatórios. Após o dia 30 os ratos foram eutanizados e suas córneas enviadas para análise histopatológica e ultra-estrutural. RESULTADOS: Dois olhos no grupo 2 foram excluídos da análise devido à infecção. Não foi encontrada diferença estatisticamente significante entre os grupos em relação à avaliação clínica. Na microscopia eletrônica de transmissão, a membrana basal nos grupos de MA liofilizada e controle foi mais contínua e homogênea em relação ao grupo da MA criopreservada. CONCLUSÕES: O processo de liofilização parece ser boa opção para a preservação da membrana amniótica humana para utilização na reconstrução da superfície ocular.