Cristina Adelaide Figueiredo

Genetic Homogeneity of Measles Viruses Associated with a Measles Outbreak, São Paulo, Brazil, 1997

During a resurgence of measles in São Paulo, Brazil, in 1997, >40,000 cases (peak incidence rate of 246/100,000 inhabitants) and 42 measles-related deaths were reported. Reverse transcriptase-polymerase chain reaction and nucleotide sequencing were used to analyze specimens from patients who had typical clinical measles infection during this outbreak and from six patients who had had measles in 1995 and 1996. Although wild-type measles viruses (genotypes D5 and D6) were present in São Paulo before this resurgence, we detected only D6 viruses. The genotype D6 viruses isolated during this outbreak had identical sequences to genotype D6 viruses isolated in other parts of Brazil and South America in 1997 and 1998, suggesting that a single chain of transmission was responsible. We also identified genotype A viruses in two vaccine-associated cases from 1995 and 1996. Our findings extend the knowledge of the circulation patterns of measles virus in South America, contributing to measles control efforts in the Americas.

Antiviral Action of Hydromethanolic Extract of Geopropolis from Scaptotrigona postica against Antiherpes Simplex Virus (HSV-1)

The studies on chemical composition and biological activity of propolis had focused mainly on species Apis mellifera L. (Hymenoptera: Apidae). There are few studies about the uncommon propolis collected by stingless bees of the Meliponini tribe known as geopropolis. The geopropolis from Scaptotrigona postica was collected in the region of Barra do Corda, Maranhão state, Brazil. The chemical analysis of hydromethanolic extract of this geopropolis (HMG) was carried out through HPLC-DAD-ESI-MS/MS and the main constituents found were pyrrolizidine alkaloids and C-glycosyl flavones. The presence of alkaloids in extracts of propolis is detected for the first time in this sample. The antiviral activity of HMG was evaluated through viral DNA quantification experiments and electron microscopy experiments. Quantification of viral DNA from herpes virus showed reduction of about 98% in all conditions and concentration tested of the HMG extract. The results obtained were corroborated by transmission electron microscopy, in which the images did not show particle or viral replication complex. The antiviral activity of C-glycosyl flavones was reported for a variety of viruses, being observed at different points in the viral replication. This work is the first report about the antiviral activity of geopropolis from Scaptotrigona postica, in vitro, against antiherpes simplex virus (HSV).

Identification of Serologic Markers for School-Aged Children With Congenital Rubella Syndrome

BACKGROUNDnCongenital rubella syndrome (CRS) case identification is challenging in older children since laboratory markers of congenital rubella virus (RUBV) infection do not persist beyond age 12 months.nnnMETHODSnWe enrolled children with CRS born between 1998 and 2003 and compared their immune responses to RUBV with those of their mothers and a group of similarly aged children without CRS. Demographic data and sera were collected. Sera were tested for anti-RUBV immunoglobulin G (IgG), IgG avidity, and IgG response to the 3 viral structural proteins (E1, E2, and C), reflected by immunoblot fluorescent signals.nnnRESULTSnWe enrolled 32 children with CRS, 31 mothers, and 62 children without CRS. The immunoblot signal strength to C and the ratio of the C signal to the RUBV-specific IgG concentration were higher (P < .029 for both) and the ratio of the E1 signal to the RUBV-specific IgG concentration lower (P = .001) in children with CRS, compared with their mothers. Compared with children without CRS, children with CRS had more RUBV-specific IgG (P < .001), a stronger C signal (P < .001), and a stronger E2 signal (P ≤ .001). Two classification rules for children with versus children without CRS gave 100% specificity with >65% sensitivity.nnnCONCLUSIONSnThis study was the first to establish classification rules for identifying CRS in school-aged children, using laboratory biomarkers. These biomarkers should allow improved burden of disease estimates and monitoring of CRS control programs.

Acute liver failure associated with rubella virus in a child.

Acute liver failure is a syndrome with a wide range of etiologic possibilities in children, but in up to 50% of the cases in the literature no diagnosis is established. This case report adds rubella virus to the list of possible causes of acute liver failure. This association was made by serologic, cell culture, molecular, histopathologic, and immunohistochemical methods.

Resurgence of measles virus in São Paulo, Brazil.

The epidemic that occurred in SP in 1997 showed the circulation of the D6 group. After this period there was a decrease in the individuals susceptible to it. Due to good strategies for the close observation of measles virus, the circulation of indigenous cases was not registered for the mentioned period. However, in 2001, 2002 and 2005, cases of an imported virus belonging to the group D5 was registered.

The antiviral effect of mollusk mucus on measles virus.

Measles is a viral disease highly contagious spread by respiratory transmission. Although infection can be controlled by vaccination, numerous cases of measles have been registered in many areas of the world, highlighting the need for additional interventions. Terrestrial gastropods exude mucus on their body surface when traveling, to protect the body from mechanical injury, desiccation or contact with harmful substances. The mucus of mollusks has been studied as a source of new natural compounds with diverse biological activities. In this study, the antiviral activity of the mucus of the land slug P.xa0boraceiensis was demonstrated inxa0vitro using Vero cells infected with measles virus. The crude sample and four fractions were tested in cultures infected with measles virus and the antiviral activity was assessed by the cytopathic effect in infected cell cultures as well as by immunofluorescence and qPCR. Fractions 39 and 50 of the mucus from P.xa0boraceiensis were analyzed by HPLC-DAD-ESI-MS/MS and infrared spectroscopy. A mixture of polyunsaturated fatty acids was found in the two fractions. A reduction in the growth of the measles virus was observed, measured by qPCR, with a protection index of 80% in Vero cells infected with measles and treated with fraction 39. Fraction 39 exhibited the best antiviral action inxa0vitro and high contents of hydroxy-tritriacontapentaenoic acid and hydroxy-pentatriacontapentaenoic acid were found in this fraction.

Infectivity of wild-type rubella virus in fibrochondrocyte cells

This study describes the rapid growth of the rubella virus in samples of a primary fibrochondrocyte cell culture with the development of a cytopathic effect (CPE), in response to infection by the rubella virus. The cells were isolated from the meniscus joint of a rabbit after enzymatic extraction and incubated at 37°C with a Dulbeccos modified Eagles medium (DMEM), supplemented with 10% fetal calf serum. A total of six clinical samples from urine, blood and cerebrospinal fluid were inoculated in the fibrochondrocyte and the cell lines of the African green monkey kidney - ATCC CCL-81 (Vero). The fibrochondrocyte cell showed CPE after 24 hours and virus growth was confirmed by immunohistochemistry and Nested PCR. The cells inoculated with samples were examined by phase contrast microscopy and showed characteristic rounding, along with bipolar and multipolar cells. The infection curve increased during the five days of observation, showing that the titers in fibrochondrocyte cells were then higher than those observed in Vero cell lines.

Intermittent rash, lymph node swelling, arthralgia and vaccinal viral detection after rubella immunization

A 26-year-old woman was immunized on August 2008 with the ubella/measles vaccine (MR, Serum Institute of India). She had een in good health previously, with no medical history of any heumatologic affection. The patient had already been vaccinated gainst rubella two years before, in an immunization campaign that ccurred in Brazilian the state of Sao Paulo, and had no adverse eactions. She inadvertently took the vaccine again in the national ampaign in 2008. Five days after the most recent vaccination, she eveloped fever, retroauricular lymph node swelling, erythemaous maculopapular rash on her trunk, back, abdomen, forearm nd ankle. On the subsequent days, she had arthralgia and a slight welling in small joints of the hands and feet. The fever resolved n ten days but the rash on her back progressed to persistent yperpigmentation, which also occurred over the joints of some ngers. Over the subsequent 18 months, the patient kept having ntermittent episodes of the arthralgia in the same joints, lymhadenopathy and erythematous macules on her arms, abdomen nd legs. The results of the laboratory tests in the first office visit ere as follows: hemoglobin, 11.9 g/dL; leukocytes, 6700/mm3

Single-dose varicella vaccine effectiveness in Brazil: A case-control study

BACKGROUNDnVaricella vaccine was introduced into the Brazilian Immunization Program in October 2013, as a single-dose schedule administered at 15u202fmonths of age. Its effectiveness had not yet been assessed in the country.nnnMETHODSnA matched case-control study was carried out in São Paulo and Goiânia (Southeast and Midwest regions, respectively), Brazil. Suspected cases, were identified through a prospective surveillance established in the study sites. All cases had specimens from skin lesion collected for molecular laboratory testing. Cases were confirmed by either clinical or PCR of skin lesions and classified as mild, moderate, and severe disease. Two neighborhood controls were selected for each case. Cases and controls were aged 15-32u202fmonths and interviewed at home. Evidence of prior vaccination was obtained from vaccination cards. Univariate and multivariate logistic regression models were used, and odds ratio and its respective 95% confidence intervals were estimated. Vaccine effectiveness was estimated by comparing de odds of having received varicella vaccine among cases and controls.nnnRESULTSnA total of 168 cases and 301 controls were enrolled. Moderate and severe illness, was found in 33.3% and 9.9% of the cases. Effectiveness of a single dose varicella vaccine was 86% (95%CI 72-92%) against disease of any severity and 93% (95%CI 82-97%) against moderate and severe disease. Out of 168 cases, 81.8% had positive PCR results for wild-type strains, and 22.0% were breakthrough varicella cases. Breakthrough cases were milder compared to non-breakthrough cases (pu202f<u202f.001).nnnCONCLUSIONSnEffectiveness of single dose varicella vaccine in Brazil is comparable to that in other countries where breakthrough varicella cases have also been found to occur. The goal of the varicella vaccination program, along with disease burden and affordability should be taken into consideration when considering the adoption of a second dose of varicella vaccine into national immunization programs.

Polyunsaturated fatty acids from Phyllocaulis boraceiensis mucus block the replication of influenza virus

Influenza viruses cause worldwide outbreaks and pandemics in humans and animals every year with considerable morbidity and mortality. The molecular diversity of secondary metabolites extracted from mollusks is a good alternative for the discovery of novel bioactive compounds with unique structures and diverse biological activities. Phyllocaulis boraceiensis is a hermaphroditic slug that exudes mucus, in which was detected hydroxy polyunsaturated fatty acids that exhibited potent antiviral activity against measles virus. The objective of this study was to evaluate this property against Influenza viruses. Cell viability and toxicity of the mucus were evaluated on Madin–Darby canine kidney (MDCK) cells by MTT assay. Antiviral activity from mucus against influenza viruses was carried out by determination of the virus infection dose and by immunofluorescence assays. The crude mucus and its fractions exhibited low cytotoxicity on MDCK cells. A significant inhibition of viral replication, reduced by the order of eight times, was observed in influenza-induced cytopathic effect. In immunofluorescence assay was observed a decrease of more than 80% of the viral load on infected MDCK cell treated with mucus and its fractions. The viral glycoproteins hemagglutinin and neuraminidase located on the surface of the virus are crucial for the replications and infectivity of the influenza virus. Some authors demonstrated that lipids, such as, polyunsaturated fatty acids exhibited multiple roles in antiviral innate and adaptive responses, control of inflammation, and in the development of antiviral therapeutics. As corroborated by other studies, hydroxy polyunsaturated fatty acids interfered with the binding of influenza virus on host cell receptor and reduced viral titers. The results obtained indicated that polyunsaturated fatty acids from P. boraceiensis crude mucus and fractions 39 exerted antiviral activity against influenza virus.