Zika virus damages the human placental barrier and presents marked fetal neurotropism

An unusually high incidence of microcephaly in newborns has recently been observed in Brazil. There is a temporal association between the increase in cases of microcephaly and the Zika virus (ZIKV) epidemic. Viral RNA has been detected in amniotic fluid samples, placental tissues and newborn and fetal brain tissues. However, much remains to be determined concerning the association between ZIKV infection and fetal malformations. In this study, we provide evidence of the transplacental transmission of ZIKV through the detection of viral proteins and viral RNA in placental tissue samples from expectant mothers infected at different stages of gestation. We observed chronic placentitis (TORCH type) with viral protein detection by immunohistochemistry in Hofbauer cells and some histiocytes in the intervillous spaces. We also demonstrated the neurotropism of the virus via the detection of viral proteins in glial cells and in some endothelial cells and the observation of scattered foci of microcalcifications in the brain tissues. Lesions were mainly located in the white matter. ZIKV RNA was also detected in these tissues by real-time-polymerase chain reaction. We believe that these findings will contribute to the body of knowledge of the mechanisms of ZIKV transmission, interactions between the virus and host cells and viral tropism.

Depression and diagnosis of neurocognitive impairment in HIV-positive patients

Neurocognitive impairment (NCI) is frequently observed in patients infected with human immunodeficiency virus (HIV) and results from the compromise of subcortical brain structures by the virus. The manifestations of NCI range from asymptomatic impairment to dementia. In addition to cognitive impairment resulting from HIV infection, other factors such as depression are associated with the loss of cognitive functions. The aim of this study was to estimate the prevalence of NCI in HIV-positive patients in a city in southern Brazil and to establish possible associations for the prevalence of NCI with HIV-related and other risk factors. This cross-sectional study of HIV-positive outpatients was conducted in a specialized care service in the city of Pelotas in Southern Brazil. Sociodemographic data and HIV-related information were collected, and all patients underwent psychiatric and neurocognitive evaluations. The prevalence of NCI among the 392 patients was 54.1% when tracked using the IHDS (International HIV Dementia Scale) and 36.2% when the IHDS was associated with a battery of complementary tests. A bivariate analysis suggested an association of NCI with gender, age, educational level, depression, current CD4 count and lowest CD4 count. The association of NCI with depression remained in the Poisson regression (PR=1.96, 95%CI=1.12-3.42). The prevalence of cognitive impairment in HIV-positive patients estimated in this study is in accordance with international and Brazilian data. Of the factors analyzed, depression showed the greatest evidence of association with neurocognitive loss. Based on our findings, the inclusion of instruments to evaluate depression in our services for patients with HIV and acquired immunodeficiency syndrome (AIDS) is recommended.

VIROLOGICAL AND SEROLOGICAL DIAGNOSIS OF RABIES IN BATS FROM AN URBAN AREA IN THE BRAZILIAN AMAZON / Diagnóstico virológico e sorológico de raiva em morcegos de uma área urbana na Amazônia Brasileira

The outbreaks of rabies in humans transmitted by Desmodus rotundus in 2004 and 2005, in the northeast of the Brazilian State of Para, eastern Amazon basin, made this a priority area for studies on this zoonosis. Given this, the present study provides data on this phenomenon in an urban context, in order to assess the possible circulation of the classic rabies virus (RABV) among bat species in Capanema, a town in the Amazon basin. Bats were collected, in 2011, with mist nets during the wet and dry seasons. Samples of brain tissue and blood were collected for virological and serological survey, respectively. None of the 153 brain tissue samples analyzed tested positive for RABV infection, but 50.34% (95% CI: 45.67-55.01%) of the serum samples analyzed were seropositive. Artibeus planirostris was the most common species, with a high percentage of seropositive individuals (52.46%, 95% CI: 52.31 52.60%). Statistically, equal proportions of seropositive results were obtained in the rainy and dry seasons (c2 = 0.057, d.f. = 1, p = 0.88). Significantly higher proportions of males (55.96%, 95% CI: 48.96-62.96%) and adults (52.37%, 95% CI: 47.35-57.39%) were seropositive. While none of the brain tissue samples tested positive for infection, the high proportion of seropositive specimens indicates that RABV may be widespread in this urban area.

Os surtos de raiva em humanos transmitida por Desmodus rotundus em 2004 e 2005 no nordeste do estado do Pará, Brasil, Amazônia Oriental, fizeram desta uma área prioritária para estudos sobre essa zoonose. Diante disso, o presente estudo fornece dados sobre esse fenômeno em contexto urbano, afim de avaliar uma possível circulação do vírus clássico da raiva (RABV) entre espécies de morcegos em Capanema, cidade localizada na bacia Amazônica. Os morcegos foram coletados em 2011, com auxílio de redes de espera durante as estações seca e chuvosa. Amostras de encéfalo e de sangue foram coletadas para o diagnóstico virológico e sorológico, respectivamente. Das 153 amostras de encéfalo analisadas, nenhuma encontrou-se positiva para infecção pelo RABV, porém, 50,34% (95% CI: 45,67-55,01) das amostras de soro analisadas estavam soropositivas. Artibeus planirostris foi a espécie mais comum, e seu percentual de indivíduos soropositivos foi bem elevado (52.46%, 95% CI: 52,31-52,60). Porções estatisticamente iguais de soropositivos foram registrados nas estações (c2 = 0,057, d.f. = 1, p = 0,88). Uma porção significativamente elevada de machos (55,96%, 95% CI: 48,96%-62,96%), e adultos (52,37%, 95% CI: 47,35%-57,39%) foram soropositivos. Apesar de nenhuma das amostras de encéfalo terem sido positivas para raiva, a alta proporção de espécimes soropositivos indica uma possível circulação do RABV nessa área urbana.

Prediction of brain atrophy using three drug scores in neuroasymptomatic HIV-infected patients with controlled viremia

ABSTRACTBACKGROUND: Despite potent antiretroviral therapy, HIV still causes brain damage. Better penetration into the CNS and efficient elimination of monocyte/macrophages reservoirs are two main characteristics of an antiretroviral drug that could prevent brain damage. The aim of our study was to assess efficacy of three antiretroviral drug scores to predict brain atrophy in HIV-infected patients.METHODS:A cross sectional study consisting of 56 HIV-infected patients with controlled viremia, who had no clinically evident neurocognitive impairment. All patients had MRI of the head. A typical T2 transversal slice was analyzed and ventricles-brain ratio (VBr) as an overall brain atrophy index was calculated. Three antiretroviral drug scores were used and correlated with VBr: 2008 and 2010 CNS penetration effectiveness scores (SCPE2008 and SCPE2010) and the recently established monocyte efficacy (SME) score. A p-value <0.05 was considered significant.RESULTS:SCPE2010 was significantly associated with VBr in both univariate (r = -0.285, p = 0.033) and multivariate (ß = -0.299, p = 0.016) regression models, while SCPE2008 was not (r = -0.141, p = 0.300 and ß = -0.156,p = 0.214). SME was associated with VBr in multivariate model only (r = -0.297, p = 0.111 andß = -0.406, p = 0.029). Age and reported duration of HIV infection were also significant predictors of overall brain atrophy in multivariate regression models.CONCLUSIONS:Although based on similar type of research, SCPE2010 is a superior drug score compared to SCPE2008. SME is an efficient drug score in determining brain damage. Both SCPE2010 and SME scores should be taken into account in preventive strategies of brain atrophy and neurocognitive impairment in HIV-infected patients.

Percepção dos graduandos de enfermagem sobre o seu envelhecimento / Percepción de los estudiantes de enfermería sobre lo proprio envejecimiento / Perception of nursing undergraduate students on self-aging

RESUMO Objetivo: analisar a percepção dos graduandos de enfermagem sobre o próprio envelhecimento. Método: pesquisa de abordagem qualitativa, realizada em agosto e setembro de 2011, com 18 graduandos de enfermagem de uma Universidade pública de Salvador (Bahia). Os depoimentos foram analisados por meio da Análise de Conteúdo. Resultados: apreendeu-se o núcleo temático: Percepção do graduando de enfermagem sobre o próprio envelhecimento e, a partir deste, emergiram duas subcategorias: A) O Não Pensar; B) O contexto influenciando no processo. Conclusão: os graduandos revelam que o envelhecimento está intrínseco ao desenvolvimento humano, e possui o vínculo familiar, a espiritualidade e atividade física como ferramentas fundamentais para um envelhecimento ativo. Entretanto, os mesmos relatam que, o modo de vida acelerado e estressante vivido na sociedade possibilita inserir hábitos considerados inadequados, como o consumo de “fast food” e álcool, que trazem influências negativas para o próprio processo de envelhecimento. .

RESUMEN Objetivo: analizar la percepción de los estudiantes de enfermería sobre su proprio envejecimiento. Método: estudio cualitativo, realizado en agosto y septiembre de 2011, con 18 estudiantes de enfermería de una universidad pública en Salvador/Bahia. Los datos fueron analizados através de análisis de contenido. Resultado: incautados el tema central: Percepción de alumnos de enfermería sobre su propio envejecimiento y de esto surgieron dos subcategorías: A) No creo; B) El contexto influye en el proceso. Conclusión: los estudiantes revelan que el envejecimiento es intrínseco al desarrollo humano, y tiene los vínculos familiares, la espiritualidad y la actividad física como herramienta clave para el envejecimiento activo. Sin embargo, el mismo informe que, debido a la forma de vida que se vive en la sociedad de ritmo rápido y estresante permite insertar hábitos considerados inadecuados, como el consumo de “comida rápida” y el alcohol y convertirse en influencias negativas para su propio proceso tuvo como objetivo analizar de los estudiantes de enfermería su propio envejecimiento. .

ABSTRACT Objective: to analyze the perceptions of nursing undergraduate students on their self-aging process. Method: qualitative study carried out between August and September, 2011 with 18 nursing undergraduate students of a public university in Salvador, Bahia. The interviews were analyzed by means of the Content Analysis method. Results: the following thematic concept was apprehended: Perceptions of nursing undergraduates on their self-aging, which generated two subcategories: A) The “don’t think about it” process; B) The context infl uencing the process. Conclusion: undergraduates reveal that the aging process is an intrinsic factor to human development. Family ties, spirituality and physical activity would be key mechanisms toward active aging. However, students also reported that their accelerated and stressed social lifestyles led to inadequate habits, such as the consumption of fast food and alcohol, which become negative infl uences in their aging process. .

Medium-term cryopreservation of rabies virus samples

Introduction The cryopreservation of rabies virus has been described in detail in the literature. To date, little information is available on the use of cryoprotective agents for cold preservation of this virus, and the available data focus only on short-term virus preservation. In this study, we investigated the medium-term cryopreservation of samples of rabies virus using different cryopreservation protocols. Methods The cryopreservation protocols for the rabies virus samples were performed at -20°C and were divided according to the variables of time and cryoprotectant type used. The laboratory tests (intracerebral inoculation of mice, viral titration and direct immunofluorescence) were performed at regular intervals (360 and 720 days) to assess the viability of the viral samples according to the different preservation techniques used. Results After 1 year of cryopreservation, the fluorescence intensity of intracellular corpuscles of the rabies virus and the median survival time of the mice differed between the positive controls and the treatments with the cryoprotectants. After 2 years, most of the samples subjected to the cryopreservation protocols (including the controls) did not produce fluorescence. However, the virus samples exposed to the cryoprotectant sucrose (68% solution) responded positively in the direct immunofluorescence assay and in the intracerebral inoculation of the mice. Conclusions Medium-term cryopreservation of the rabies virus inactivates the viral sample. However, the cryoprotectant agent sucrose (68%) produces a preservative effect in cryopreserved rabies virus samples. .

HAM/TSP: association between white matter lesions on magnetic resonance imaging, clinical and cerebrospinal fluid findings / HAM/TSP: associação entre lesões de substância branca à ressonância magnética, achados clínicos e do líquido cefalorraquidiano

OBJECTIVE: To investigate the association between clinical data, white matter lesions and inflammatory cerebrospinal fluid (CSF) findings in HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP). METHOD: We studied brain and cervical spinal cord on magnetic resonance imaging (MRI) and CSF examinations of 28 Brazilian HAM/TSP patients. RESULTS: The majority of patients had severe neurological incapacity with EDSS median of 6.5 (3-8). The brain MRI showed white matter lesions (75%) and atrophy (14%). The preferential brain location was periventricular. Cervical demyelination lesions occurred in 11% of the cases, and cervical atrophy in 3.5%. One patient had enhancement lesions on T1 cervical spinal cord MRI. Cases with spinal cord lesions had signs of acute CSF inflammation. The brain white matter lesions predominated in the patients with higher age. CONCLUSION: Our data suggest that an active inflammatory process is associated with the cervical spinal cord lesions in HAM/TSP. The brain abnormalities are not related to the clinical picture of HAM/TSP.

OBJETIVO: Analisar a associação entre aspectos clínicos, lesões de substância branca e reação inflamatória aguda no líquido cefalorraquidiano (LCR) na mielopatia associa ao HTLV-1 (HAM/TSP). MÉTODO: Foram estudadas ressonâncias magnéticas (RM) do encéfalo/medula espinhal cervical e exame do LCR de 28 pacientes com HAM/TSP. RESULTADOS: A maioria dos pacientes apresentava grave incapacidade neurológica, com EDSS 6,5 (3-8). A RM revelou lesões da substância branca (75%) com predominância periventricular e atrofia cortical (14%). Lesões desmielinizantes cervicais ocorreram em 11% dos casos e atrofia em 3,5%. Um paciente apresentou lesão cervical na T1 com captação de contraste. Sinais de inflamação aguda no LCR ocorreram em situações de lesão da medula espinhal cervical. As alterações de substância branca do encéfalo predominaram nos indivíduos com maior faixa etária. CONCLUSÃO: Nossos achados sugerem que processo inflamatório com atividade clínica na HAM/TSP está associado a lesões da medula espinhal cervical. As anormalidades da substância branca encefálicas não são relacionadas ao quadro clínico de HAM/TSP.

Comparison of four diagnostic methods for detecting rabies viruses circulating in Korea

It is essential to rapidly and precisely diagnose rabies. In this study, we evaluated four diagnostic methods, indirect fluorescent antibody test (FAT), virus isolation (VI), reverse transcriptase polymerase chain reaction (RT-PCR), and rapid immunodiagnostic assay (RIDA), to detect rabies in animal brain homogenates. Out of the 110 animal brain samples tested, 20 (18.2%) were positive for rabies according to the FAT. Compared to the FAT, the sensitivities of VI, RT-PCR, and RIDA were 100, 100, and 95%, respectively. The specificities of VI, RT-PCR and RIDA were found to be 100, 100, and 98.9%, respectively. Rabies viruses circulating in Korea were isolated and propagated in murine neuroblastoma (NG108-15) cells with titers ranging from 101.5 to 104.5 TCID50/mL. Although the RIDA findings did not completely coincide with results obtained from FAT, VI, and RT-PCR, RIDA appears to be a fast and reliable assay that can be used to analyze brain samples. In summary, the results from our study showed that VI, RT-PCR, and RIDA can be used as supplementary diagnostic tools for detecting rabies viruses in both laboratory and field settings.

Antemortem diagnosis of human rabies in a veterinarian infected when handling a herbivore in Minas Gerais, Brazil / Diagnóstico ante-mortem de raiva humana em médico veterinário infectado por manipulação de herbívoro, Minas Gerais, Brasil

The Ministry of Health's National Human Rabies Control Program advocates pre-exposure prophylaxis (PEP) for professionals involved with animals that are at risk of contracting rabies. We report an antemortem and postmortem diagnosis of rabies in a veterinarian who became infected when handling herbivores with rabies. The antemortem diagnosis was carried out with a saliva sample and a biopsy of hair follicles using molecular biology techniques, while the postmortem diagnosis used a brain sample and conventional techniques. The veterinarian had collected samples to diagnose rabies in suspect herbivores (bovines and caprines) that were subsequently confirmed to be positive in laboratory tests. After onset of classic rabies symptoms, saliva and hair follicles were collected and used for antemortem diagnostic tests and found to be positive by RT-PCR. Genetic sequencing showed that the infection was caused by variant 3 (Desmodus rotundus), a finding confirmed by tests on the brain sample. It is essential that professionals who are at risk of infection by the rabies virus undergo pre-exposure prophylaxis. This study also confirms that molecular biology techniques were used successfully for antemortem diagnosis and therefore not only allow therapeutic methods to be developed, but also enable the source of infection in human rabies cases to be identified accurately and quickly.

O Programa Nacional de Controle da Raiva Humana do Ministério da Saúde preconiza o esquema profilático pré-exposição (PEP) para profissionais envolvidos com animais expostos ao risco de contraírem raiva. O presente trabalho relata o diagnóstico de raiva (ante e post-mortem) em veterinário infectado por manipulação de herbívoros raivosos. O diagnóstico laboratorial ante-mortem foi efetuado a partir da saliva e biópsia de folículo piloso, utilizando técnicas de biologia molecular e o post-mortem a partir do tecido cerebral e de técnicas convencionais. O médico veterinário coletou amostras para diagnóstico de raiva em herbívoros (bovinos e caprinos) suspeitos que, posteriormente, foram confirmados positivos em laboratório. Após a apresentação dos sintomas clássicos de raiva e realizadas as provas de diagnóstico ante-mortem com saliva e folículo piloso, ambas as amostras apresentaram resultados positivos pelo nested-RT-PCR. O sequenciamento genético revelou que a infecção se deu pela variante 3 do Desmodus rotundus, resultados estes confirmados com a amostra do cérebro. É indispensável que profissionais expostos ao risco de infecção pelo vírus da raiva realizem a profilaxia pré-exposição. Ressalta-se, também, que as técnicas de biologia molecular apresentaram bons resultados para a realização de diagnóstico ante-mortem, propiciando o desenvolvimento de métodos terapêuticos, e determinando com precisão e rapidez a fonte de infecção dos casos de raiva humana.

A bovine herpesvirus 5 recombinant defective in the thymidine kinase (TK) gene and a double mutant lacking TK and the glycoprotein E gene are fully attenuated for rabbits

Bovine herpesvirus 5 (BoHV-5), the agent of herpetic meningoencephalitis in cattle, is an important pathogen of cattle in South America and several efforts have been made to produce safer and more effective vaccines. In the present study, we investigated in rabbits the virulence of three recombinant viruses constructed from a neurovirulent Brazilian BoHV-5 strain (SV507/99). The recombinants are defective in glycoprotein E (BoHV-5gEÄ), thymidine kinase (BoHV-5TKÄ) and both proteins (BoHV-5gEÄTKÄ). Rabbits inoculated with the parental virus (N = 8) developed neurological disease and died or were euthanized in extremis between days 7 and 13 post-infection (pi). Infectivity was detected in several areas of their brains. Three of 8 rabbits inoculated with the recombinant BoHV-5gEÄ developed neurological signs between days 10 and 15 pi and were also euthanized. A more restricted virus distribution was detected in the brain of these animals. Rabbits inoculated with the recombinants BoHV-5TKÄ (N = 8) or BoHV-5gEÄTKÄ (N = 8) remained healthy throughout the experiment in spite of variable levels of virus replication in the nose. Dexamethasone (Dx) administration to rabbits inoculated with the three recombinants at day 42 pi did not result in viral reactivation, as demonstrated by absence of virus shedding and/or increase in virus neutralizing titers. Nevertheless, viral DNA was detected in the trigeminal ganglia or olfactory bulbs of all animals at day 28 post-Dx, demonstrating they were latently infected. These results show that recombinants BoHV-5TKÄ and BoHV-5gEÄTKÄ are attenuated for rabbits and constitute potential vaccine candidates upon the confirmation of this phenotype in cattle.

Experimental infection of chickens, ducks and quails with the highly pathogenic H5N1 avian influenza virus

Highly pathogenic avian influenza viruses (HPAIV) of the H5N1 subtype have spread since 2003 in poultry and wild birds in Asia, Europe and Africa. In Korea, the highly pathogenic H5N1 avian influenza outbreaks took place in 2003/2004, 2006/2007 and 2008. As the 2006/2007 isolates differ phylogenetically from the 2003/2004 isolates, we assessed the clinical responses of chickens, ducks and quails to intranasal inoculation of the 2006/2007 index case virus, A/chicken/Korea/IS/06. All the chickens and quails died on 3 days and 3-6 days post-inoculation (DPI), respectively, whilst the ducks only showed signs of mild depression. The uninoculated chickens and quails placed soon after with the inoculated flock died on 5.3 and 7.5 DPI, respectively. Both oropharyngeal and cloacal swabs were taken for all three species during various time intervals after inoculation. It was found that oropharyngeal swabs showed higher viral titers than in cloacal swabs applicable to all three avian species. The chickens and quails shed the virus until they died (up to 3 to 6 days after inoculation, respectively) whilst the ducks shed the virus on 2-4 DPI. The postmortem tissues collected from the chickens and quails on day 3 and days 4-5 and from clinically normal ducks that were euthanized on day 4 contained the virus. However, the ducks had significantly lower viral titers than the chickens or quails. Thus, the three avian species varied significantly in their clinical signs, mortality, tissue virus titers, and duration of virus shedding. Our observations suggest that duck and quail farms should be monitored particularly closely for the presence of HPAIV so that further virus transmission to other avian or mammalian hosts can be prevented.

Antigenic and genetic characterization of the first rabies virus isolated from the bat Eumops perotis in Brazil

Although the main transmitters of rabies in Brazil are dogs and vampire bats, the role of other species such as insectivorous and frugivorous bats deserves special attention, as the rabies virus has been isolated from 36 bat species. This study describes the first isolation of the rabies virus from the insectivorous bat Eumops perotis. The infected animal was found in the city of Ribeirão Preto, São Paulo. The virus was identified by immunofluorescence antibody test (FAT) in central nervous system (CNS) samples, and the isolation was carried out in N2A cell culture and adult mice. The sample was submitted to antigenic typing using a panel of monoclonal antibodies (CDC/Atlanta/USA). The DNA sequence of the nucleoprotein gene located between nucleotides 102 and 1385 was aligned with homologous sequences from GenBank using the CLUSTAL/W method, and the alignment was used to build a neighbor-joining distance-based phylogenetic tree with the K-2-P model. CNS was negative by FAT, and only one mouse died after inoculation with a suspension from the bat's CNS. Antigenic typing gave a result that was not compatible with the patterns defined by the panel. Phylogenetic analysis showed that the virus isolated segregated into the same cluster related to other viruses isolated from insectivorous bats belonging to genus Nyctinomops ssp. (98.8 percent nucleotide identity with each other).

No Brasil, embora os principais transmissores da raiva sejam cães e morcegos hematófagos, o papel de outras espécies, tais como morcegos insetívoros e frugívoros, merece atenção especial, uma vez que o vírus da raiva já foi isolado em 36 espécies de morcegos. Este estudo descreve o primeiro isolamento do vírus da raiva em um morcego insetívoro Eumops perotis. O animal infectado foi encontrado na cidade de Ribeirão Preto, São Paulo. O vírus foi identificado pelo teste de imunofluorescência direta (IFD) em amostras de sistema nervoso central (SNC), e o isolamento foi realizado em cultura de células N2A e em camundongos adultos. A amostra foi submetida à tipificação antigênica, utilizando um painel de oito anticorpos monoclonais (CDC/Atlanta/USA). A seqüência de DNA do gene da nucleoproteína, localizada entre os nucleotídeos 102 a 1385, foi alinhada com seqüências homólogas presentes no GenBank, usando o método CLUSTAL/W e o alinhamento foi utilizado para a construção da árvore filogenética de distância "neighbor-joining" com o modelo K-2-P. O SNC testado foi negativo por IFD, e somente um camundongo morreu após inoculação com a suspensão do SNC do morcego. A tipificação antigênica apresentou resultado não-compatível com os padrões definidos pelo painel. A análise filogenética mostrou que o vírus isolado segregou no mesmo grupo relacionado com outros vírus isolados de morcegos insetívoros, gênero Nyctinomops ssp. (98,8 por cento de identidade de nucleotídeos entre elas).

Vírus rábico em morcego Nyctinomops laticaudatus na Cidade do Rio de Janeiro, RJ: isolamento, titulação e epidemiologia / Rabies virus in Nyctinomops laticaudatus bats in the City of Rio de Janeiro: isolation, titration and epidemiology

Apresenta-se o primeiro relato de raiva em morcego da espécie Nyctinomops laticaudatus, na Cidade do Rio de Janeiro, RJ. Foram realizados isolamento e titulação viral em diferentes tecidos, encontrando-se altos títulos no cérebro e glândulas salivares. A ocorrência de raiva em uma espécie pouco freqüente neste estado sugere que a doença pode ser mais prevalente do que aparenta.

The first case report of rabies in bats of the species Nyctinomops laticaudatus, in the city of Rio de Janeiro City, is presented. Virus isolation and titration were performed in different tissues, and high titers were found in the brain and salivary glands. Rabies occurrence in such an infrequent species in this state suggests that the disease may be more prevalent than it appears to be.

Application of immunofluorescence and sandwich ELISA with double-antibodies in detection of human rabies / 法医学杂志

OBJECTIVE@#To explore the application of immunofluorescence and sandwich ELISA with double-antibodies in detection of human rabies.@*METHODS@#The cerebrum, cerebellum, brainstem, and hippocampus of four patients died of rabies identified by clinical diagnosis were collected and kept in freezer at -70 degrees C or in formaldehyde solution separately. The rat brain tissue infected by CVS strain of rabies virus was used as a positive control and the brain tissue of a patient died of acute pancreatitis was used as a negative control.@*RESULTS@#Rabies virus was detected in the tissues kept in freezer at -70 degrees C and the positive control but was not detected in the tissues kept in formaldehyde solution and the negative control.@*CONCLUSION@#Immunofluorescence and Sandwich ELISA with double-antibodies could be used in detection of human rabies. The samples should be kept in deep frozen temperature condition instead of in formaldehyde solution.

Resonancia Magnética de médula espinal y cerebro en el correlato clínico de la paraparesia espástica progresiva que se asocia al virus humano linfotrópico tipo-I (HTLV-I) / Brain and spinal cord magnetic resonance imaging in spastic paraparesis associated to human T-lymphotropic virus

Background: The spastic paraparesis associated to HTLV-1 causes degenerative pyramidal tract lesions of the spinal cord and affects cortical-nuclear connections in the brain. Aim: To report the findings of magnetic resonance imaging in patients with spastic paraparesis. Material and methods: A magnetic resonance imaging of the brain and spinal cord was performed in 30 patients (24 females), mean age and evolution of 56 and 12 years respectively, with a clinical and virological diagnosis of tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM). Results: No patient had abnormal signals in the spinal cord parenchyma. However, an atrophy of the dorsal segment was observed in 87 percent of patients. Patients with the highest degree of atrophy showed a higher degree of functional impairment. Eleven patients had spinal cord conus atrophy, associated to neurogenic bladder or impotency. In 80 percent of patients, hyperintense subcortical white matter images in DP, T2 and Flair, mostly bi frontal, were detected. In half of them, small rounded and isolated images were observed. In the other half, eight or more images, generally larger and occasionally confluent, were found. Ten of 12 patients with confluent brain lesions showed different degrees of cognitive impairment. No patient had lesions in the corpus callosus, periventricular white matter, pons, medulla oblongata or cerebellum. Conclusions: Most patients with tropical spastic paraparesis have alterations in brain or spinal cord magnetic resonance imaging. The magnetic resonance lesions are concordant with functional impairment. The characteristics of the imaging in TSP/HAM patients can be helpful in the differential diagnosis of patients with paraparesis.

Dexamethasone-induced reactivation of bovine herpesvirus type 5 latent infection in experimentally infected rabbits results in a broader distribution of latent viral DNA in the brain

Bovine herpesvirus type 5 (BHV-5) is a major agent of meningoencephalitis in cattle and establishes latent infections mainly in sensory nerve ganglia. The distribution of latent BHV-5 DNA in the brain of rabbits prior to and after virus reactivation was studied using a nested PCR. Fifteen rabbits inoculated intranasally with BHV-5 were euthanized 60 days post-inoculation (group A, N = 8) or submitted to dexamethasone treatment (2.6 mg kg-1 day-1, im, for 5 days) and euthanized 60 days later (group B, N = 7) for tissue examination. Two groups of BHV-1-infected rabbits (C, N = 3 and D, N = 3) submitted to each treatment were used as controls. Viral DNA of group A rabbits was consistently detected in trigeminal ganglia (8/8), frequently in cerebellum (5/8), anterior cerebral cortex and pons-medulla (3/8) and occasionally in dorsolateral (2/8), ventrolateral and posterior cerebral cortices, midbrain and thalamus (1/8). Viral DNA of group B rabbits showed a broader distribution, being detected at higher frequency in ventrolateral (6/7) and posterior cerebral cortices (5/7), pons-medulla (6/7), thalamus (4/7), and midbrain (3/7). In contrast, rabbits inoculated with BHV-1 harbored viral DNA almost completely restricted to trigeminal ganglia and the distribution did not change post-reactivation. These results demonstrate that latency by BHV-5 is established in several areas of the rabbit's brain and that virus reactivation leads to a broader distribution of latent viral DNA. Spread of virus from trigeminal ganglia and other areas of the brain likely contributes to this dissemination and may contribute to the recrudescence of neurological disease frequently observed upon BHV-5 reactivation.

Yellow fever virus isolated from a fatal post vaccination event: an experimental comparative study with the 17DD vaccine strain in the Syrian hamster (Mesocricetus auratus) / Vírus da febre amarela isolado de acidente pós-vacinal fatal: estudo experimental comparativo com a amostra vacinal 17DD em Syrian hamsters (Mesocricetus auratus)

In order to investigate the pathogenicity of the virus strain GOI 4191 that was isolated from a fatal adverse event after yellow fever virus (YFV) vaccination, an experimental assay using hamsters (Mesocricetus auratus) as animal model and YFV 17DD vaccine strain as virus reference was accomplished. The two virus strains were inoculated by intracerebral, intrahepatic and subcutaneous routes. The levels of viremia, antibody response, and aminotransferases were determined in sera; while virus, antigen and histopathological changes were determined in the viscera. No viremia was detected for either strain following infection; the immune response was demonstrated to be more effective to strain GOI 4191; and no significant aminotransferase levels alterations were detected. Strain GOI 4191 was recovered only from the brain of animals inoculated by the IC route. Viral antigens were detected in liver and brain by immunohistochemical assay. Histothological changes in the viscera were characterized by inflammatory infiltrate, hepatocellular necrosis, and viral encephalitis. Histological alterations and detection of viral antigen were observed in the liver of animals inoculated by the intrahepatic route. These findings were similar for both strains used in the experiment; however, significant differences were observed from those results previously reported for wild type YFV strains.

Visando investigar a possível patogenicidade do vírus isolado (GOI 4191) de um evento adverso fatal pela vacinação antiamarílica, realizou-se um ensaio experimental em Syrian hamsters (Mesocricetus auratus), usando-se a cepa vacinal 17DD como parâmetro. As amostras virais foram inoculadas por via intracerebral, intra-hepática e subcutânea. Nos soros foram determinados níveis de viremia, resposta imune e aminotransferases, e nas vísceras a presença de vírus, antígeno e lesões teciduais. Não se detectou viremia para as duas amostras, a resposta imune foi maior para GOI 4191, e as aminotransferases não apresentaram alterações compatíveis com danos hepáticos. Nos animais inoculados por via intracerebral o vírus foi recuperado somente a partir do cérebro, sendo o antígeno viral detectado, por imuno-histoquímica, no cérebro e fígado. Infiltrado inflamatório e corpúsculos acidófilos foram observados no fígado e lesões tipo encefalite viral no sistema nervoso central. Alterações histológicas e antígeno viral foram observados, também, no fígado dos animais infectados por via intra-hepática, e ausentes naqueles inoculados por via subcutânea. Os resultados foram similares para as duas amostras testadas, entretanto distintos daqueles relatados na literatura para cepas silvestres do vírus amarílico.

Distribution of Adenoviral Vector in Brain after Intravenous Administration

The delivery of transgenes to the central nervous system (CNS) can be a valuable tool to treat CNS diseases. Various systems for the delivery to the CNS have been developed; vascular delivery of viral vectors being most recent. Here, we investigated gene transfer to the CNS by intravenous injection of recombinant adenoviral vectors, containing green fluorescence protein (GFP) as a reporter gene. Expression of GFP was first observed 6 days after the gene transfer, peaked at 14 days, and almost diminished after 28 days. The observed expression of GFP in the CNS was highly localized to hippocampal CA regions of cerebral neocortex, inferior colliculus of midbrain, and granular cell and Purkinje cell layers of cerebellum. It is concluded that intravenous delivery of adenoviral vectors can be used for gene delivery to the CNS, and hence the technique could be beneficial to gene therapy.

Experimental study of mouse cytomegalovirus infected mice / 华中科技大学学报（医学）（英德文版）

In order to investigate the human cytomegalovirus (HCMV) infection, the mouse cytomegalovirus (MCMV) infected mice were experimentally studied. 6 to 8 week old female BALB/C mice with immunosuppression were selected to undergo the MCMV inoculations: intracranial inoculation and peritoneal inoculation. MCMV of the infected mice in various organs and tissues were detected by using beta-gal staining and in situ nucleic acid hybridization assay. The pathological changes were observed in HE staining paraffin-embedded sections. It was found that all the MCMV infected mice showed the retardation of growth and development, and feather looseness. Both intracranial inoculation of 10(4) PFU viruses or peritoneal inoculation of 10(6) PFU viruses resulted in the pathological changes, to some extent, of various organs and tissues in the mice. The pathological changes in liver were consistent with the amount of beta-gal staining positive cells, indicating the liver lesions were mainly caused by viral proliferation. It was also found that the viruses in the immunosuppressed mice subjected to intracranial inoculation could spread to whole body organs, while the viruses in the immunosuppressed mice subjected to intrapeitoneal inoculation couldn't spread to the brain, suggesting blood-brain barrier could prevent the virus from spreading to the brain.