Possible treatments for sporadic Creutzfeldt-Jakob disease: a systematic review

m o Sistema Nervoso, na qual ocorre um acúmulo anormal de uma proteína conhecida como Prion Scrapie (PrPsc). A PrPsc é uma mutação de uma proteína do tecido cerebral chamada Prion Celular Normal (PrPc) e o acúmulo de PrPsc causa deficiências espongiformes. A Doença de Creutzfeldt-Jakob (CJD) é uma das várias Doenças Priônicas e sua forma esporádica (CJDe) é a mais prevalente. Considerando a progressão implacável e rápida da doença, o desfecho invariavelmente letal e a escassez de opções terapêuticas para as Doenças Priônicas, especialmente a DCJ, tratamentos eficazes tornam-se necessários. Assim, o objetivo desta revisão sistemática é analisar e identificar opções terapêuticas voltadas para a DCJe. Esta pesquisa foi realizada nas bases de dados Pubmed e Cochrane com os termos Mesh “Sporadic Creutzfeldt-Jakob disease” e “Treatment”, e o booleano “AND”. Foram selecionados quatro artigos em inglês, que mais se alinhavam com os objetivos do trabalho. Como resultado desta pesquisa, chegou-se à conclusão de que dois medicamentos foram bem tolerados pelos pacientes, porém não proporcionaram uma melhor qualidade de vida e nem alcançaram maior longevidade. Outros medicamentos estão sendo testados em pacientes adultos, alguns em ensaios clínicos apresentaram boa eficácia, minimizando déficits cognitivos em pacientes com DCJ. No entanto, os estudos necessitam de um maior número de ensaios em uma grande quantidade de pacientes para melhor conclusão que aborde o tratamento desta doença grave e fatal.Prion Diseases are rare and fatal diseases that compromises the Nervous System, in which occurs an abnormal accumulation of a protein known as Prion Scrapie (PrPsc). PrPsc is a mutation of a brain tissue protein called Normal Cellular Prion (PrPc) and the accumulation of PrPsc causes spongiform deficiencies. Creutzfeldt-Jakob Disease (CJD) is one of the various Prion Diseases and its sporadic form (sCJD) is the most prevalent. Considering the relentless and rapid progression of the disease, the invariably lethal outcome and the scarcity of therapeutic options for Prion Diseases, especially sCJD, effective treatments become necessary. Thus, the objective of this systematic review is to analyze and identify therapeutic options aimed at sCJD. This research was carried out in the Pubmed and Cochrane databases with the Mesh Terms “Sporadic Creutzfeldt-Jakob disease” and “Treatment”, and the boolean “AND”. Four articles in English were selected, that most aligned ones with the work targets. As the outcome of this research, came to a conclusion that two drugs were well tolerated by the patients, nevertheless they did not deliver a better life quality neither achieved an extended life span. Other drugs are being tested in adult patients, some in clinical trials exhibited good efficacy, minimizing cognitive deficits in CJD patients. However, the studies require a greater number of trials in a wide amount of patients for better conclusion that addresses the treatment of this severe and fatal disease

Detection of hantavirus in bats from remaining rain forest in São Paulo, Brazil

Background\ud The significant biodiversity found in Brazil is a potential for the emergence of new zoonoses. Study in some places of the world suggest of the presence to hantavirus in tissues of bats. Researches of hantavirus in wildlife, out rodents, are very scarce in Brazil. Therefore we decided to investigate in tissues of different species of wild animals captured in the same region where rodents were detected positive for this virus. The present work analyzed ninety-one animals (64 rodents, 19 opossums, and 8 bats) from a region of the Atlantic forest in Biritiba Mirin City, São Paulo State, Brazil. Lungs and kidneys were used for RNA extraction.\ud \ud Findings\ud The samples were screened for evidence of hantavirus infection by SYBR-Green-based real-time RT-PCR. Sixteen samples positive were encountered among the wild rodents, bats, and opossums. The detection of hantavirus in the lungs and kidneys of three marsupial species (Micoureus paraguayanus, Monodelphis ihering, and Didelphis aurita) as well in two species of bats (Diphylla ecaudata and Anoura caudifer) is of significance because these new hosts could represent an important virus reservoirs.This work was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo - FAPESP. The field support and animals identification was made at Museum of Zoology, Universidade de Sao Paulo, USP and Universidade Bandeirante, UNIBAN, Brazil

Passive hemaglutination technique for ant dengue human immunoglobulin detection

A dengue é uma doença febril, exantemática aguda considerada a mais importante arbovirose que afeta o homem. O diagnóstico laboratorial da dengue é realizado após suspeita clínica e as técnicas imunoenzimáticas, principalmente ELISA, são as mais difundidas, detectando anticorpos das classes IgM e IgG. Este trabalho teve como objetivo o desenvolvimento de uma técnica de hemaglutinação passiva de fácil execução, rápida e eficiente na detecção de anticorpos antidengue em sangue humano total. As próprias hemácias do paciente foram utilizadas como antígeno figurado, sendo o anti- soro contra hemácias humanas produzido em coelhos e purificado por precipitação e cromatografia. As imunoglobulinas G obtidas foram digeridas com o uso de papaína para a obtenção de fragmentos Fab. Após a separação, os fragmentos foram ligados às partículas virais de dengue, previamente purificadas em gradiente de densidade contínuo de tartarato de sódio e potássio, utilizando o glutaraldeído como agente acoplador. Demonstrou-se que a estratégia de conjugação dos fragmentos Fab à partícula viral é factível com uso de glutaraldeído, contudo, há necessidade do desenvolvimento de estudos para a verificação do grau de eficiência de conjugação. O método de conjugação adotado não impediu os fragmentos Fabs de se ligarem às hemácias e nem os anticorpos antidengue de reconhecerem seus epítopos na partícula viral. A utilização de anticorpos monoclonais, ou sistemas de expressão para proteínas virais assim como para fragmentos de anticorpos facilitariam as etapas de montagem do conjugado, além de otimizar o funcionamento da técnica. Em conclusão, a detecção de anticorpos antidengue em amostras de soro e hemácias foi possível a partir do método proposto.Dengue is an acute febrile illness considered the most important arthropod-borne viral disease affecting humans. Currently most laboratory diagnoses of dengue are made after some clinical observations and use of immunoenzimatic techniques, notably ELISA can reliably detect small amounts of IgM and IgG classes of antibodies and have deserved worldwide acceptance. However, ELISA is a somewhat laborious, time-consuming technique and requires a expensive equipment. The present work was carried out to develop a feasible, inexpensive, quick and efficient passive hemaglutination technique for detection of anti-dengue antibodies in human whole blood, using the patient's blood cells as figured antigen. Antiserum against human erythrocytes was produced in rabbits and the IgG was purified by precipitation and chromatographic methods. IgG was then fractioned by papain digestion to yield Fab fragments. After separation, these fragments were linked by glutaraldehyde to the dengue viral particles successfully purified following a protocol which included centrifugation through a sodium and potassium tartarate density- gradient. However, the glutaraldehyde-mediated step need further work to establish an efficient degree of conjugation. The method did not alter either the Fab binding to erythrocytes or the recognition of viral epitopes by anti-dengue antibodies. Possibly, the use of monoclonal antibodies or a suitable system for protein expression could improve the conjugation step and help to optimize the routine testing of samples. In conclusion, a simple technique has been demonstrated to be suitable for detection of anti-dengue antibodies in samples of human blood or erythrocytes plus serum.Conselho Nacional de Desenvolvimento Científico e Tecnológic

Molecular characterization of arboviruses isolated from mosquitoes fauna (Diptera: nematocera) Rondonia state (western brazilian Amazon).

Rondônia apresenta área com rica diversidade de artrópodes, porém pouco se conhece sobre a transmissão de arbovírus por estas espécies. O presente trabalho visou detectar arbovírus, por meio da RT-PCR e da Duplex RT-PCR, nas espécies de dipteros coletados no Estado, bem como caracterizá-los pela reação de sequenciamento. A RT-PCR e a Duplex RT-PCR detectaram as suspensões dos vírus Mayaro e Oropouche até 104 e 101 TCID50/mL, respectivamente, porém o vírus Dengue 2 em pools contendo menos de três mosquitos infectados foi negativa. O controle endógeno foi detectado em 66,8 % das amostras, sendo que, em pools contendo entre um e três mosquitos, a detecção foi aproximadamente metade da detecção nos pools contendo entre 11 e 15. Em 0,66 % dos pools foi encontrado o vírus Oropouche e em outros 0,66 %, o vírus Cacipacoré. O vírus Oropouche foi detectado em Coquillettidia sp. e Deinocerites sp., enquanto o vírus Cacipacoré foi encontrado em Anopheles sp. e Culex sp. As técnicas possibilitaram a detecção dos arbovírus pesquisados nos pools coletados em Rondônia.The Rondônia state has an area with rich arthropods diversity although the knowledge about the arboviruses transmition for these species is poor. The present work aimed to detect arboviruses through RT-PRC and RT-PCR Duplex in the diptera species collected in the region as well as their characterization through the sequence reaction. The RT-PRC and RT-PCR Duplex detected the Mayaro and Oropouche virus suspensions until 104 e 101 TCID50/mL respectively, although it was negative for the Dengue 2 virus in pools containing less than three infected mosquitoes. The endogenous control was detected in 66,8 % of samples and from pools containing from one to three mosquitoes the detection rate was approximately half from that obtained from pools with 11 to 15 mosquitoes. Oropouche virus was found in 0,66 % of pools and Cacipacore virus also in 0,66 % of pools. Oropouche virus was detected in Coquillettidia sp. and Deinocerites sp. while Cacipacoré virus was found in Anopheles sp. and Culex sp. The techniques allowed the detection of examined arboviruses in the pools collected from Rondonia

Cacipacore virus as an emergent mosquito-borne Flavivirus

Abstract INTRODUCTION: Cacipacore virus (CPCV), a possible bird-associated flavivirus, has yet to be detected in mosquitoes. Our purpose is examining CPCV in mosquitoes from the Amazon region of Brazil. METHODS: Approximately 3,253 Culicidae (grouped into 264 pools) were collected from the Amazon region during 2002-2006 and analyzed using a Flavivirus genus-specific reverse transcription- polymerase chain reaction followed by nested polymerase chain reaction assay and by nucleotide sequencing of amplicons. RESULTS: Nucleotide sequences from five mosquito samples showed high similarity to the those of CPCV originally isolated in the Amazon region. CONCLUSIONS: This is the first report of CPCV-infected mosquitoes which has implications on the arbovirus maintenance in nature and transmission to man