Genética forense: fundamentos e aplicações / Forensic genetics: fundamentals and applications

Por um longo período, a elucidação de crimes foi desempenhada apenas por testemunhas. Tal panorama sofreu uma grande revolução com o uso das impressões digitais como ferramenta forense, a papiloscopia. Desde então, inúmeras técnicas foram criadas para o desvendamento de crimes, sendo que a Genética Forense merece um papel de destaque. A invenção da Reação em Cadeia da Polimerase (PCR, do inglês Polymerase Chain Reaction) tornou possível desenvolver ferramentas de genética forense que permitem tanto investigações de rotina muito informativas quanto investigações especiais, ainda mais avançadas, em casos relativos a crimes, paternidade, identificação de vítimas de desastres, entre outras. A presente revisão aborda o conceito de variabilidade genética aplicado à técnica de PCR para a obtenção de perfis genéticos, no âmbito da genética forense, além de contextualizar sobre a importância da criação do Banco Nacional de Perfis Genéticos para as investigações criminais

A PERCEPÇÃO DO TRATAMENTO COM ÓLEO DE CANNABIS EM UM PACIENTE PORTADOR DE AUTISMO:: UM RELATO DE CASO

O Transtorno de Espectro Autista, é uma doença que acomete crianças em idade de desenvolvimento, principalmente no sexo masculino. Sua manifestação inclui impacto nas relações sociais e habilidade de comunicação, associado a comportamentos estereotipados. Seu tratamento consiste em abordagem multidisciplinar, necessitando polimedicação. Nesse sentido, a Cannabis sativa tem ganhado destaque pelos efeitos farmacológicos sobre doenças neurológicas e psiquiátricas, tornando-se uma opção no tratamento do Autismo. Este estudo, com abordagem qualitativa, teve como objetivo fazer um relato da percepção dos pais de uma criança, portadora de Autismo, sobre o uso do óleo de cannabis. O principal relato dos pais foi a satisfação na melhora do desenvolvimento geral da criança, evolução na independência durante a realização das necessidades fisiológicas, nas habilidades motoras, comportamentais e de interação social. Destacou-se a dificuldade na aquisição do óleo de cannabis no Brasil. Os efeitos da Cannabis no Autismo se dão possivelmente pela regulação da neurotransmissão, através da modulação de receptores canabinóides, com atuação sob demanda, que ocorre através de um mecanismo de transmissão retrógrada, caracterizando esse sistema como neuromodulador. Portanto, este relato indicou diversos benefícios da Cannabis sobre o TEA, sendo necessários avanços nas pesquisas a fim de elucidar seus mecanismos de atuação específicos nessa doença

Paracoccidioidomycosis in wild and domestic animals

A paracoccidioidomicose é a micose sistêmica mais prevalente na América Latina, principalmente no Brasil. Apesar de existirem vários estudos sobre o diagnóstico e patologia dessa micose, pouco se conhece sobre a ecologia do agente etiológico Paracoccidioides brasiliensis. Seu habitat ainda não foi determinado, mas provavelmente o fungo vive saprofiticamente no solo. Há também pouca informação quanto a participação de outras espécies de animais na eco-epidemiologia do fungo e os isolamentos obtidos de animais como morcego e pingüim não foram reprodutíveis. Até o presente, o fungo tem sido isolado com maior freqüência do tatu, Dasypus novencinctus. Estudos soroepidemiológicos demonstraram que a infecção é freqüente em cães, principalmente da área rural e recentemente foi relatado o primeiro caso de paracoccidiodomicose natural em cão. O papel de outras espécies de animais na eco-epidemiologia do fungo ainda não está esclarecido.Paracoccidiodomycosis is the most prevalent systemic mycosis in Latin América, mainly in Brazil. Although many studies have been done about the pathology and diagnosis of this mycosis little is known about the eco-epidemiology of the etiological agent Paracoccidioides brasiliensis. The fungus habitat also is unknown although is thought that the fungus lives as a saprobe in soil. The participation of other animal species in P. brasiliensis ecology is poorly understood and fungus isolation from animals as bat and penguin were not reproducible. The armadillos Dasypus novencinctus probably play a role in the fungus eco-epidemiology taking into account that several isolates were obtained from these animals. Seroepidemiologic studies have shown that dogs are infected by P. brasiliensis mainly in rural areas and recently was reported the first case o natural paracoccidioidomycosis in dogs. The role of other animal species in the eco-epidemiology of P. brasiliensis remains doubtful

MicroRNA-100 Acts as a Tumor Suppressor in Human Bladder Carcinoma 5637 Cells

Bladder carcinoma is one of the most common tumors in the world and, despite the therapy currently available, most of the patients relapse. Better understanding of the factors involved in disease pathogenesis would provide insights for the development of more effective strategies in treatment. Recently, differential miRNA expression profiles in bladder urothelial carcinomas identified miR-100 down-regulation and miR-708 up-regulation among the most common alterations, although the possible influence of these miRNAs in the control of basic mechanisms in bladder tumors has not been addressed. In this context, the present study aimed to evaluate the in vitro effects of miR-100 forced expression and miR-708 inhibition in the bladder carcinoma cell line 5637. Our results showed that overexpression of miR-100 significantly inhibited growth when compared to controls at both times tested (72 and 96 hours, p<0.01) with a maximum effect at 72 hours reducing proliferation in 29.6 %. Conversely, no effects on cell growth were observed after inhibition of miR-708. MiR-100 also reduced colony formation capacity of 5637 cells by 24.4%. No alterations in cell cycle progression or apoptosis induction were observed. The effects of miR-100 on growth and clonogenicity capacity in 5637 cells evince a possible role of this miRNA in bladder carcinoma pathogenesis. Further studies are necessary to corroborate our findings and examine the potential use of this microRNA in future therapeutic interventions

DIFFERENTIAL MICRORNAS EXPRESSIONS ARE ASSOCIATED WITH RELAPSE AND TP53 P.R377H MUTATION IN PEDIATRICS ADRENOCORTICAL TUMORS

5961070107

Protection conferred by heterologous vaccination against tuberculosis is dependent on the ratio of CD4(+)/CD4(+) Foxp3(+) cells

CD4(+) Foxp3(+) regulatory T cells inhibit the production of interferon-?, which is the major mediator of protection against Mycobacterium tuberculosis infection. In this study, we evaluated whether the protection conferred by three different vaccines against tuberculosis was associated with the number of spleen and lung regulatory T cells. We observed that after homologous immunization with the 65 000 molecular weight heat-shock protein (hsp 65) DNA vaccine, there was a significantly higher number of spleen CD4(+) Foxp3(+) cells compared with non-immunized mice. Heterologous immunization using bacillus Calmette Guerin (BCG) to prime and DNA-hsp 65 to boost (BCG/DNA-hsp 65) or BCG to prime and culture filtrate proteins (CFP)-CpG to boost (BCG/CFP-CpG) induced a significantly higher ratio of spleen CD4(+)/CD4(+) Foxp3(+) cells compared with non-immunized mice. In addition, the protection conferred by either the BCG/DNA-hsp 65 or the BCG/CFP-CpG vaccines was significant compared with the DNA-hsp 65 vaccine. Despite the higher ratio of spleen CD4(+)/CD4(+) Foxp3(+) cells found in BCG/DNA-hsp 65-immunized or BCG/CFP-CpG-immunized mice, the lungs of both groups of mice were better preserved than those of DNA-hsp 65-immunized mice. These results confirm the protective efficacy of BCG/DNA-hsp 65 and BCG/CFP-CpG heterologous prime-boost vaccines and the DNA-hsp 65 homologous vaccine. Additionally, the prime-boost regimens assayed here represent a promising strategy for the development of new vaccines to protect against tuberculosis because they probably induce a proper ratio of CD4(+) and regulatory (CD4(+) Foxp3(+)) cells during the immunization regimen. In this study, this ratio was associated with a reduced number of regulatory cells and no injury to the lungs.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2007/02695-5

IGF2 and IGF1R in pediatric adrenocortical tumors: roles in metastasis and steroidogenesis

Deregulation of the IGF system observed in human tumors indicates a role in malignant cell transformation and in tumor cell proliferation. Although overexpression of the IGF2 and IGF1R genes was described in adrenocortical tumors (ACTs), few studies reported their profiles in pediatric ACTs. In this study, the IGF2 and IGF1R expression was evaluated by RT-qPCR according to the patient’s clinical/pathological features in 60 pediatric ACT samples, and IGF1R protein was investigated in 45 samples by immunohistochemistry (IHC). Whole transcriptome and functional assays were conducted after IGF1R inhibition with OSI-906 in NCI-H295A cell line. Significant IGF2 overexpression was found in tumor samples when compared with non-neoplastic samples (P1μM) showed limited reduction of cell viability, suggesting that OSI-906 alone could not be a suitable therapy to abolish carcinoma cell growth.236481493CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPSem informação2010/07020-

MicroRNA profile of pediatric pilocytic astrocytomas identifies two tumor-specific signatures when compared to non-neoplastic white matter

PurposesPilocytic astrocytoma (PA) is a low-grade neoplasm frequently found in childhood. PA is characterized by slow growth and a relatively good prognosis. Genetic mechanisms such as activation of MAPK, BRAF gene deregulation and neurofibromatosis type 1 (NF1) syndrome have been associated with PA development. Epigenetic signature and miRNA expression profile are providing new insights about different types of tumor, including PAs.MethodsIn the present study we evaluated global miRNA expression in 16 microdissected pediatric PA specimens, three NF1-associated PAs and 11 cerebral white matter (WM) samples by the microarray method. An additional cohort of 20 PAs was used to validate by qRT-PCR the expression of six miRNAs differentially expressed in the microarray data.ResultsUnsupervised hierarchical clustering analysis distinguished one cluster with nine PAs, including all NF1 cases and a second group consisting of the WM samples and seven PAs. Among 88 differentially expressed miRNAs between PAs and WM samples, the most underexpressed ones regulate classical pathways of tumorigenesis, while the most overexpressed miRNAs are related to pathways such as focal adhesion, P53 signaling pathway and gliomagenesis. The PAs/NF1 presented a subset of underexpressed miRNAs, which was also associated with known deregulated pathways in cancer such as cell cycle and hippo pathway.ConclusionsIn summary, our data demonstrate that PA harbors at least two distinct miRNA signatures, including a subgroup of patients with NF1/PA lesions1412373382CNPQ - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPESP – Fundação de Amparo à Pesquisa Do Estado De São Paulo471885/2013-42010/07020-