Performance of non-formalin fixed paraffin embedded samples in hybrid capture and amplicon next-generation sequencing panels.

BACKGROUND: Genomic profiling using next-generation sequencing (NGS) is fundamental for driving prognostic and therapy in cancer. Formalin-fixed paraffin embedded (FFPE) tissue is the widely used material, whereas non-FFPE may represent an alternative. However, studies comparing the NGS performance of non-FFPE materials to FFPE are still lacking in the literature. The objective of this study was to characterize in non-FFPE preparations the nucleic acid yield and NGS performance on both a capture-based and an amplicon-based NGS platform. NGS quality metrics obtained from non-FFPE preparations were compared to FFPE. METHODS: We analyzed the cellularity and nucleic acid yield in 111 tumors from non-FFPE preparations. In addition, comprehensive hybrid capture panel sequencing metrics obtained from DNA and RNA libraries were compared between independent non-FFPE and FFPE samples. A paired comparison between non-FFPE and FFPE samples was performed to analyze concordance in mutant allele detection using an amplicon panel. RESULTS: The mean target coverage from DNA libraries was 2× higher in non-FFPE samples than in FFPE. The detection of exogenous DNA was 2.5× higher in non-FFPE than in FFPE. Conversely, a lower performance was observed in non-FFPE RNA libraries in comparison to FFPE DNA libraries with no impact in minimum standard cutoffs. The variant allele detection in non-FFPE was found to be comparable to that of FFPE tumor samples in matched samples. CONCLUSIONS: Non-FFPE was demonstrated to be a suitable material for DNA and RNA library preparations using a comprehensive NGS panel. This is the first study reporting library quality metrics according to the TSO500 analysis pipeline.

Revisión bibliográfica de resultados obtenidos utilizando la Técnica de Masquelet en lesiones complicadas con osteomielitis: a propósito de un caso clínico / Bibliographic review of results obtained using Masquelet's technique in complicated lesions with osteomyelitis: apropos of a clinical case

SUMMARY: The Masquelet technique or membrane induction is considered new in many ways, born under the need to seek therapeutic options in patients with extensive bone lesions. Since this technique was proposed, hopeful and reproducible results have been reported to different centers throughout the world. That is why in this work we seek to collect information from different authors and their case reports, in addition to presenting a case handled in the O'higgins region with this technique. OBJECTIVES: To review the literature regarding general results in bone consolidation in cases similar to the one exposed, in addition to exposing the Masquelet Technique as management in a patient with extensive bone loss, due to a firearm wound. METHODS: descriptive observational study, in addition to a systematic review in databases such as PubMed/MEDLINE, Elsevier, Cochrane and manually through the Internet in journals and public bodies. This work seeks to collect information from different authors and their case reports, in addition to delving into the technique itself, evaluating its indications, contraindications and protocol to follow. The patient's signature of an informed consent was requested, which is explicitly voluntary, in which he authorizes the review of his file, his background and the use of images and / or x-rays pertinent to the research. RESULTS: Inclusion and exclusion criteria were defined to analyze the characteristics of the selected articles. We present the clinical case of a 27-year-old male patient who suffers high-energy injury by firearm in the middle third of the right leg with exposure and loss of musculoskeletal tissue of 12 cm in diameter, polyfragmentary fracture of the proximal third of tibia and fibula, initially damage control is performed which is complicated by presenting osteomyelitis in said limb. It is handled with Masquelet technique. The induction time was approximately 4 months, after the second surgical time the lesion is consolidated in three months showing results similar to the literature studied.

Revisión bibliográfica de Fasciolosis biliar: propósito de un caso / Bibliographic review of biliary fasciolosis: purpose of a case

The biliary pathology, is undoubtedly one of the most frequent surgical pathologies in Chile, statistical data establish the incidence of biliary pathology in over 20 years 30% of women have biliary lithiasis, of these 10% will present some complication as it is coledocolithiasis. On the other hand, hepatic fascioliasis or dystomatosis is a disease caused by hepatic fasciola, or duela, sabuaypé or Distomun hepaticum, corresponds to a flatworm, trematode of the digenea class, with affinity to stay in liver tissue, which when performing the biological cycle of the parasite can be the cause of choledocholithiasis, cholecystitis, cholangitis, pancreatitis, among others. We present a clinical case of chronic intracholedocianfasciolitis diagnosed by ERCP3,4. METHODS: Descriptive observational study, in addition to a systematic review in databases such as Pub-Med/MEDLINE, Elsevier, Cochrane and manually through the Internet in journals and public bodies. This work seeks to collect information from different authors regarding its incidence, management and established treatments. RESULTS: Inclusion and exclusion criteria were defined to analyze the characteristics of the selected articles. We present the clinical case of a 47-year-old female patient, with a history of hypertension, type 2 diabetes mellitus and egg allergy who went to the Emergency Department due to a picture characterized by epigastralgia of 3 days of evolution. During endoscopic retrograde cholangio-pancreatography (ERCP), 2 suggestive images of lyte are seen inside that finally result in 2 apparent live parasites.

Novel Mutations in Pilomatrixoma, CTNNB1 p.s45F, and FGFR2 p.s252L: A Report of Three Cases Diagnosed by Fine-Needle Aspiration Biopsy, with Review of the Literature.

Pilomatrixoma (calcifying epithelioma of Malherbe) is an uncommon benign skin appendageal tumor that differentiates toward hair matrix cells. It is misdiagnosed in up to 75% of cases by nondermatologists. Although the histopathological findings are well recognized and characteristic, diagnosis by fine-needle aspiration biopsy may be quite challenging. Several reports have emphasized the challenges in cytodiagnosis of pilomatrixoma, leading to a false-positive diagnosis. The lesions may show avidity for fludeoxyglucose on positron emission tomography/computed tomography scan, raising concern of a possible malignant neoplasm. CTNNB1 mutations have been reported in a high percentage of pilomatrixomas. Expression of ß-catenin, the protein encoded by CTNNB1, is also frequently observed. To determine if routine cytological specimens can be successfully used to perform additional investigation and support or confirm the diagnosis in three cases of pilomatrixoma, we performed molecular analysis and immunohistochemistry to search for CTNNB1 mutation and ß-catenin, respectively. ß-Catenin positivity by immunohistochemistry was observed in basaloid cells in all three cases. Exon 3 mutations in CTNNB1 were detected in all cases. In addition, we detected a fibroblast growth factor receptor 2 (FGFR2) mutation in one of the cases. We reviewed the literature and present the clinical and morphological characteristics that must be considered along with other findings to accurately achieve the correct diagnosis, in correlation with the results of the ancillary technique. In conclusion, routine cytological specimens can be successfully used to perform additional investigations and support cytodiagnosis in difficult cases.

Cómo entender la experiencia de personas que viven con VIH: implicaciones para la clínica y la investigación / How to Understand the Experience of Persons Living with HIV: Implications for Clinical Practice and Research / Como entender a experiência de pessoas que vivem com hiv: implicações para a clínica e para a pesquisa

Objetivo: las personas que viven con una enfermedad crónica experimentan procesos complejos de transformación personal para adaptarse a su nueva condición de salud. El objetivo del artículo es develar el significado de vivir con VIH en personas que fueron diagnosticadas en etapa de sida. Materiales y métodos: estudio cualitativo descriptivo en el que se realizaron entrevistas en profundidad a 15 personas que viven con VIH (PVVIH) en etapa de sida, que se atienden en el sistema público de salud chileno. Resultados: del análisis de contenido emergieron seis dimensiones del significado de vivir con la enfermedad: cambios personales posteriores al diagnóstico; apoyo social y acompañamiento, resignación a la adquisición del VIH; percepción de discriminación / rechazo; revelación del diagnóstico; cuidar a otros de la adquisición del VIH. Conclusiones: los hallazgos orientan a los profesionales de la salud sobre las estrategias que se deben utilizar en la atención directa de las personas que viven con VIH con el fin de otorgar un cuidado integral considerando el apoyo para el desarrollo de estrategias de autocuidado y la revelación del diagnóstico a su núcleo cercano, para reducir los costos psicosociales asociados a la enfermedad.

Objective: Those who live with chronic disease undergo complex processes of personal transformation to adapt to their new health condition. The aim of this article is to show what it means to live with HIV for persons who have been diagnosed with AIDS. Materials and methods: This is a descriptive qualitative study based on 15 people who are living with HIV/AIDS and being treated through the Chilean public health system. Results: The content analysis showed six dimensions of what it means to live with the disease: personal changes after the diagnosis, social support and accompaniment, being resigned to having contracted HIV, perceived discrimination / rejection, disclosure of the diagnosis, and protecting others from contracting HIV. Conclusion: The findings can provide guidance to health professionals on the strategies that should be used in providing direct care to persons living with HIV. In this respect, the goal is to offer comprehensive care, taking into account support for the development of self-care strategies and disclosure of the diagnosis to the patient's family and close friends, so as to reduce the psycho-social costs of the disease.

Objetivo: as pessoas que vivem com uma doença crônica experimentam processos complexos de transformação pessoal para se adaptar a sua nova condição de saúde. O objetivo deste artigo é revelar o significado de viver com HIV em pessoas que foram diagnosticadas em etapa de Aids. Materiais e métodos: estudo qualitativo descritivo no qual se realizaram entrevistas em profundidade com 15 pessoas que vivem com o HIV (PVHIV) em etapa de Aids, que se atendem no sistema público de saúde chileno. Resultados: da análise de conteúdo emergiram seis dimensões do significado de viver com a doença: mudanças pessoais posteriores ao diagnóstico; apoio social e acompanhamento, resignação à aquisição do HIV; percepção de discriminação ou rejeição; revelação do diagnóstico; cuidar a outros sobre a aquisição do HIV. Conclusões: as descobertas orientam os profissionais da saúde sobre as estratégias que se devem utilizar no atendimento direto das pessoas que vivem com o HIV, com o objetivo de outorgar um cuidado integral que considere o apoio para o desenvolvimento de estratégias de autocuidado e a revelação do diagnóstico a seu núcleo de pessoas próximas, para reduzir os custos psicossociais associados à doença.

Ras gene mutation is not related to tumour invasion during rat tongue carcinogenesis induced by 4-nitroquinoline 1-oxide.

BACKGROUND: The aim of this study was to investigate whether mutations in the genes H-ras and K-ras were related to the mechanism of invasion as a result of the immunoexpression of H-Ras, Ki-67, alpha-smooth muscle actin (SMA) and vascular endothelial growth factor (VEGF) during 4-nitroquinoline 1-oxide (4NQO)-induced rat tongue carcinogenesis. METHODS: Male Wistar rats were distributed into three groups of 10 animals each and treated with 50 ppm 4NQO solution through their drinking water for 4, 12 and 20 weeks. Ten animals were used as negative control. RESULTS: Although no histopathological abnormalities were induced in the epithelium after 4 weeks of carcinogen exposure, Ki-67 was overexpresssed in the 'normal' oral epithelium. In pre-neoplastic lesions at 12 weeks following carcinogen exposure, the levels of Ki-67 were increased (P < 0.05) when compared to negative control. Ki-67, alpha-SMA and VEGF were also overexpressed in squamous cell carcinomas induced after 20 weeks of treatment with 4NQO. No significant statistical differences (P > 0.05) were found in H-ras protein expression for all experimental periods evaluated that corresponded to normal oral mucosa, hyperplasia, dysplasia and squamous cell carcinomas. In the same way, no mutations in H-ras or K-ras genes were found. CONCLUSIONS: Our results support the idea that expression of Ki-67 plays a crucial role during malignant transformation being closely related to neoplastic conversion of the oral mucosa cells. However, it seems that mutations in the ras genes are not involved to experimental tongue carcinogenesis induced by 4NQO.

Doenças hematológicas associadas ao eritrovírus / Hematologic diseases associated with eritrovírus

O eritrovírus infecta células precursoras eritroides, determinando a interrupção temporária da eritropoese. Neste contexto, é importante o conhecimento das principais doenças hematológicas que podem estar associadas à presença do vírus, principalmente quando estão presentes em condições mórbidas, tais como nas anemias hemolíticas hereditárias. Este trabalho tem como objetivo relatar as principais doenças hematológicas que cursam com a infecção pelo eritrovírus B19.

Erythroviruses infect precursor erythroid cells, determining a temporary disruption of erythropoiesis. Thus, knowledge of the main hematological diseases that may be associated with the virus is important, especially when they are present in morbid conditions, such as in hereditary hemolytic anemia. This paper aims at reporting the main hematological diseases that are associated with erythrovirus infections.

Motociclistas e uso de álcool: estudo dos acidentes fatais no município de São Paulo (PAP)

O aumento da acidentalidade no trânsito está intimamente relacionado ao uso do álcool. Quando se trata de motociclistas, o uso do álcool é um importante fator de risco devido à vulnerabilidade física a que está sujeito o seu condutor. O propósito do estudo foi caracterizar a provável relação entre o uso de álcool e a morte de motociclistas com base nos dados de vítimas necropsiadas pelas Equipes de Perícias Médico-Legais do município de São Paulo entre janeiro e dezembro de 2006. As variáveis analisadas foram: idade, sexo, período e dia da semana em que ocorreu o acidente e alcoolemia de 241 vítimas fatais. O sexo masculino apresentou predominância na amostra estudada (94,2%). Houve presença de alcoolemia positiva em 46,5% das vítimas e a faixa de alcoolemia predominante foi a de 0,6 – 1,5 g/L (gramas de álcool por litro de sangue). A média de idade encontrada foi de 27,5 ± 9,13 anos. O período do dia em que ocorreu a maior porcentagem de vítimas fatais alcoolizadas foi entre 0h01 a 6 horas da manhã, no qual 63,89% das vítimas estavam alcoolizadas. Os resultados obtidos revelam a ingestão de álcool por aproximadamente metade dos motociclistas analisados, ressaltando a relevância do uso desta substância na ocorrência de acidentes envolvendo motociclistas em um nível quase três vezes maior do que o limite estabelecido no Código de Trânsito Brasileiro vigente (0,6 g/L). Tal fato evidencia a necessidade da implantação de políticas públicas de prevenção e fiscalização voltadas a essa classe de motoristas.

Papel da célula endotelial em neoplasias malignas hematológicas / The role of endothelial cells in hematologic malignancies

A neoformação vascular constitui evento essencial para que ocorra o desenvolvimento humano. Nesta fase, o sistema vascular sofre expansão. Células endoteliais maduras (ECs) irão compor a parede dos vasos, enquanto células endoteliais oriundas da medula óssea (MO) migram para locais de neoformação vascular com subseqüente diferenciação em células endoteliais maduras. Processos fisiológicos e patológicos como câncer têm a angiogênese como importante componente. Estudos correlacionam a angiogênese com a agressividade tumoral em alguns tipos de tumores incluindo hematológicos, o que ressalta a importância da terapia antiangiogênica como alvo de pesquisas cada vez mais profundas. Neste artigo de revisão enfocamos o papel da angiogênese nas neoplasias hematológicas com ênfase em possíveis alvos terapêuticos.

Vascular neoformation is an essential event in the development of the human body. In this phase vascular networks undergo expansion. Mature endothelial cells constitute the structure of vessel walls so endothelial cells from bone marrow migrate to sites of vascular neoformation with subsequent differentiation as mature endothelial cells (endothelial progenitor cells). Physiopathological and pathological processes such as cancer use angiogenesis as an important component for growth. Studies correlate angiogenesis with aggressive tumors for some types of tumors, including hematological malignancies, thereby highlighting the importance of anti-angiogenic therapy.

Presença de bactérias em instrumentais e superfícies do ambiente clínico odontológico / Presence of bacteria in instruments and surfaces of the clinic odontologic ambient

Este estudo teve como objetivo avaliar os riscos de contaminaçäo a partir da presença de bactérias em instrumentais e superfícies do ambiente clínico odontológico, supostamente apropriados para o uso do atendimento ao paciente. Foram coletadas, pela Vigilância Sanitária, 250 amostras de 49 consultórios odontológicos particulares, em diferentes bairros do município do Rio de Janeiro, e avaliados bioquimicamente e morfologicamente pelo Laboratório Central Noel Nutels. Os resultados constataram a presença de bactérias nos instrumentais e superfícies. Foi concluído que há fragilidade nas medidas de proteçäo e falhas na cadeia asséptica, o que representa risco à saúde dos pacientes e profissionais

Por que a odontologia se transformou numa profissäo de mulheres? / Why has dentistry become a womens's profession?

Este estudo teve por objetivo identificar os fatores que levaram a Odontologia a transformar-se numa profissäo com predomínio de profissionais do sexo feminino. Analisamos a identidade feminina nos aspectos biológicos, psicossociais e sua relaçäo com a reproduçäo da força de trabalho. Em conclusäo, podemos afirmar que o processo de transformaçäo deve-se a um conjunto de fatores econômicos/culturais e relaciona-se à busca pela igualdade de direitos sociais