Anti-DENV-NS1 monoclonal antibody for the differential histopathological diagnosis of hemorrhagic fever caused by dengue

Dengue is the most prevalent arboviral disease in humans in tropical and subtropical regions, especially in urban areas, and can cause major epidemics. Although a self-limiting illness, it may sometimes have serious hemorrhagic manifestations, and the outcome of dengue hemorrhagic fever has similar clinical manifestations as in other infections, which could result in death. Therefore, autopsy procedures are required under certain circumstances such as in hemorrhagic fevers, sometimes to confirm or to clarify the diagnosis that may have epidemiological consequences. Normally, the Immunohistochemistry Laboratory of the Pathology Center of Adolfo Lutz Institute receives autopsy samples from different hospitals in Sao Paulo State to confirm a previous diagnosis, especially hemorrhagic fever of infectious etiology. For this diagnosis, we have been using a mouse polyclonal antibody to dengue virus that often does not provide a clear conclusion, because of background staining or no relevant immunostaining, which hampers the histopathological analysis. Accordingly, in the present study, anti-DENV-NS1 monoclonal antibody (4H2) was tested to determine its accuracy in immunohistochemical analysis. Twenty-four autopsy cases of hemorrhagic febrile syndrome showing histopathological alterations compatible with dengue disease were studied: twenty cases were confirmed by RT-PCR for DENV-2 and in four by RT-PCR for yellow fever virus. Samples from autopsied cases of deaths caused by other infectious diseases (two meningitis C and two severe acute respiratory syndrome caused by influenza A H1N1) were included as negative control cases. Positive immunostaining for DENV-NS1 was detected in 16/20 (80%) liver samples and 11/15 (73%) spleen samples from autopsied hemorrhagic dengue patients, whereas the polyclonal antibody detected DENV antigens in 12/20 (60%) liver and in 6/15 (40%) spleen samples from the same cases. Positive results were not obtained with liver biopsy samples from yellow fever or Neisseria meningitides and Flu-A cases. 4H2 mAb recognizes the native protein of the four DENV serotypes in infected cells and did not cross-react with native ZIKV- or CHKV-infected cells by immunohistochemical assay, so it is a useful tool for differential histopathological conclusion of acute febrile hemorrhagic deaths.

The role of pregnant women with rash in the Zika virus sentinel surveillance

INTRODUCTION: The state of São Paulo has been monitoring cases of microcephaly and pregnant women presenting with acute rash, through CeVeSP. METHODS: This was a descriptive study focusing on pregnant women with rash and the outcome of their pregnancy, based on the notifications through the CeVeSP. RESULTS: During 2016, 2,209 cases of pregnant women with rash were reported and investigated. Of these, 36.6% were confirmed. Of the pregnant women who tested positive for ZIKV, 6.4% did not have a favorable outcome. CONCLUSIONS Our results allowed the characterization of pregnant women exposed to ZIKV and the outcome of pregnancy.

The role of pregnant women with rash in the Zika virus sentinel surveillance

Abstract INTRODUCTION: The state of São Paulo has been monitoring cases of microcephaly and pregnant women presenting with acute rash, through CeVeSP. METHODS: This was a descriptive study focusing on pregnant women with rash and the outcome of their pregnancy, based on the notifications through the CeVeSP. RESULTS: During 2016, 2,209 cases of pregnant women with rash were reported and investigated. Of these, 36.6% were confirmed. Of the pregnant women who tested positive for ZIKV, 6.4% did not have a favorable outcome. CONCLUSIONS Our results allowed the characterization of pregnant women exposed to ZIKV and the outcome of pregnancy.

Avaliação do número de células somáticas do leite de bovinos da raça jersey durante o primeiro mês de lactação / Evaluation of somatic cell number in milk from jersey cows during the first month of lactation

Com a finalidade de estabelecer os valores de referência da contagem de células somáticas do leite de bovinos da raça Jersey, criados no Estado de São Paulo, durante o primeiro mês de lactação, bem como avaliar a influência da fase colostral, examinaram-se 418 amostras de leite, provenientes de quartos mamários sadios e sem crescimento bacteriano. O leite foi colhido assepticamente antes da ordenha e a contagem de células somáticas foi determinada por citometria de fluxo e pelo Teste do CMT. Demonstrou-se a significativa influência do primeiro mês de da lactação sobre a contagem de células. Verificou-se que a transição da secreção de colostro para leite em relação ao número de células somáticas está finalizado a partir do 15º dia de lactação, sendo recomendado a adoção dos seguintes valores de referência: no colostro, obtidos nas primeiras 24 horas de lactação entre 472.405 e 2.003.921 células/ml, no período compreendido do 2º ao 15º dia de lactação valores entre 103.920 e 1.298.361 células/ml e a partir do 15º dia de lactação entre 37.714 e 205.549 células/ml.

With the aim to establish reference values of the number somatic cell of milk from Jersey cows, raised in the State of São Paulo, during the first month of lactation, as well as to evaluate the influence of the colostral phase, 418 milk samples were examined obtained from healthy mammary glands without bacterial growth. Milk samples were collected aseptically before milking and number somatic cell determined by electronic counting and by California Mastitis Test. was demonstrated the significant influence the somatic cells in the first month of lactation. It was found that the transition of the colostrum secretion for milk in respect to somatic cells count have been finished in the 15º day of lactation, being recommended the adoption of the following reference values: in the colostrum, getting in the first of 24 hours of lactation between 472.405 and 2.003.921cells/ml, in the understood period from 2º until 15º day of lactation values between 103.920 and 1.298.361 cells/ml and starting from 15 day of lactation between 37.714 and 205.549 cells/ml.