GBV-C/HGV and HIV-1 coinfection

An interesting interaction pattern has been found between HIV-1 and GBV-C/HGV, resulting in protection against progression to AIDS. The mechanisms involved in this interaction remain to be clarified. We examined the current knowledge concerning this coinfection and developed hypotheses to explain its effects. A better understanding of this interaction could result in new concepts, which may lead to new strategies to control HIV-1 replication and progression to AIDS.Federal University of São PauloPró-Sangue FoundationUNIFESPSciEL

SARS-CoV-2 and the COVID-19 disease: a mini review on diagnostic methods

Coronavirus disease 2019 (COVID-19) is an infectious disease initially reported in China and currently worldwide dispersed caused by a new coronavirus (SARS-CoV-2 or 2019-nCoV) affecting more than seven million people around the world causing more than 400 thousand deaths (on June 8th, 2020). The diagnosis of COVID-19 is based on the clinical and epidemiological history of the patient. However, the gold standard for COVID-19 diagnosis is the viral detection through the amplification of nucleic acids. Although the quantitative Reverse-Transcription Polymerase Chain Reaction (RT-PCR) has been described as the gold standard for diagnosing COVID-19, there are several difficulties involving its use. Here we comment on RT-PCR and describe alternative tests developed for the diagnosis of COVID-19

Factors associated with incomplete vaccination and negative antibody test results for measles, mumps, and hepatitis A among children followed in the MINA-BRAZIL cohort

Vaccination coverage has been dropping in Brazil and other countries. In addition, immune responses after vaccination may not be homogeneous, varying according to sociodemographic and clinical factors. Understanding the determinants of incomplete vaccination and negative antibody test results may contribute to the development of strategies to improve vaccination effectiveness. In this study, we aimed to investigate the frequency of vaccine adherence, factors associated with incomplete vaccination for measles, mumps, rubella (MMR) and hepatitis A, and factors associated with the seronegative test results for measles, mumps and hepatitis A at 2 years of age. This was a population-based cohort that addressed health conditions and mother/infant nutrition in Cruzeiro do Sul city, Brazil. Vaccination data were obtained from official certificates of immunization. The children underwent blood collection at the two-year-old follow-up visit; the samples were analyzed using commercially available kits to measure seropositivity for measles, mumps, and hepatitis A. We used modified Poisson regression models adjusted for covariates to identify factors associated with incomplete vaccination and negative serology after vaccination. Out of the 825 children included in the study, adherence to the vaccine was 90.6% for MMR, 76.7% for the MMRV (MMR + varicella), and 74.9% for the hepatitis A vaccine. For MMR, after the adjustment for covariates, factors associated with incomplete vaccination included: white-skinned mother; paid maternity leave; raising more than one child; lower number of antenatal consultations; and attending childcare. For hepatitis A, the factors included: white-skinned mother and not having a cohabiting partner. The factors with statistically significant association with a negative antibody test result included: receiving Bolsa Familia allowance for measles and mumps; incomplete vaccination for measles; and vitamin A deficiency for mumps. Strategies to improve the efficiency of vaccine programs are urgently needed. These include improvements in communication about vaccine safety and efficacy, and amplification of access to primary care facilities, prioritizing children exposed to the sociodemographic factors identified in this study. Additionally, sociodemographic factors and vitamin A deficiency may impact the immune responses to vaccines, leading to an increased risk of potentially severe and preventable diseases

Anti-Zika virus activity of several abietane-type ferruginol analogues

[EN] Abietane diterpenoids are naturally occurring plant metabolites with a broad spectrum of biological effects including antibacterial. antileishmanial, antitumor, antioxidant, as well as antiinfiammatory activities. Recently, we found that some analogues of natural ferruginol (2) actively inhibited dengue virus 2 (DENV-2) replication. Due to the similarity with DENY, we envisaged that abietane diteipenoids would also be active against Zika virus (ZIKV). Six selected semi-synthetic abietane derivatives of (+)-dehydroabietylatnine (3) were tested. Cytotoxicity was determined by Mn' assay in Vero cells. In vitro anti-ZIKV (clinical isolate. imT17) activity was evaluated by plaque assay. Interestingly, these molecules showed potential as anti-ZIKV agents, with EC50 values ranging from 0.67 to 18.57 mu M. and cytotoxicity (CC50 values) from 256 to 35.09 mu M. The 18-Oxoferruginol (8) (EC50 = 2.60 mu M, SI = 13.51) and 12-nitro-N-benzoyldehydroabietylamine (9) (ECG 0.67 mu M, SI = 3.82) were the most active compounds, followed by 12-hydroxy-N-tosyldehydroabietylamine (7) (EC50 = 3.58 mu M, SI = 3.20) and 12-hydroxy-N,N-phthaloyldehydroabietylamine (5) (EC50 = 7.76 mu M, SI = 1.23). To the best of our knowledge, this is the first report on anti-Zika virus properties of abietanes.Financial support from the Universitat Politecnica de Valencia, under a cooperation "ADSIDEO" research grant (AD1902), is gratefully acknowledged. We are also gratefully to the financial support of the Sao Paulo Research Foundation (FAPESP), grants No 2013/01690-0, No 2019/03859-9, and FAPESP Scholarship No 2013/017029 to FTGS.Sousa, FTG.; Nunes, C.; Malta Romano, C.; Cerdeira Sabino, E.; González-Cardenete, MA. (2020). Anti-Zika virus activity of several abietane-type ferruginol analogues. Revista do Instituto de Medicina Tropical de São Paulo. 62:1-4. https://doi.org/10.1590/S1678-9946202062097S1462Newman, D. J., & Cragg, G. M. (2020). Natural Products as Sources of New Drugs over the Nearly Four Decades from 01/1981 to 09/2019. Journal of Natural Products, 83(3), 770-803. doi:10.1021/acs.jnatprod.9b01285Hanson, J. R., Nichols, T., Mukhrish, Y., & Bagley, M. C. (2019). Diterpenoids of terrestrial origin. Natural Product Reports, 36(11), 1499-1512. doi:10.1039/c8np00079dGonzález, M. A. (2015). Aromatic abietane diterpenoids: their biological activity and synthesis. Natural Product Reports, 32(5), 684-704. doi:10.1039/c4np00110aBrandt, C. W., & Neubauer, L. G. (1939). 221. Miro resin. Part I. Ferruginol. Journal of the Chemical Society (Resumed), 1031. doi:10.1039/jr9390001031González, M. A., Clark, J., Connelly, M., & Rivas, F. (2014). Antimalarial activity of abietane ferruginol analogues possessing a phthalimide group. Bioorganic & Medicinal Chemistry Letters, 24(22), 5234-5237. doi:10.1016/j.bmcl.2014.09.061Balasubramanian, A., Teramoto, T., Kulkarni, A. A., Bhattacharjee, A. K., & Padmanabhan, R. (2017). Antiviral activities of selected antimalarials against dengue virus type 2 and Zika virus. Antiviral Research, 137, 141-150. doi:10.1016/j.antiviral.2016.11.015Roa-Linares, V. C., Brand, Y. M., Agudelo-Gomez, L. S., Tangarife-Castaño, V., Betancur-Galvis, L. A., Gallego-Gomez, J. C., & González, M. A. (2016). Anti-herpetic and anti-dengue activity of abietane ferruginol analogues synthesized from (+)-dehydroabietylamine. European Journal of Medicinal Chemistry, 108, 79-88. doi:10.1016/j.ejmech.2015.11.009González, M. A., & Pérez-Guaita, D. (2012). Short syntheses of (+)-ferruginol from (+)-dehydroabietylamine. Tetrahedron, 68(47), 9612-9615. doi:10.1016/j.tet.2012.09.055Dea-Ayuela, M. A., Bilbao-Ramos, P., Bolás-Fernández, F., & González-Cardenete, M. A. (2016). Synthesis and antileishmanial activity of C7- and C12-functionalized dehydroabietylamine derivatives. European Journal of Medicinal Chemistry, 121, 445-450. doi:10.1016/j.ejmech.2016.06.004Cory, A. H., Owen, T. C., Barltrop, J. A., & Cory, J. G. (1991). Use of an Aqueous Soluble Tetrazolium/Formazan Assay for Cell Growth Assays in Culture. Cancer Communications, 3(7), 207-212. doi:10.3727/095535491820873191Chattopadhyay, D., Sarkar, M. C.-, Chatterjee, T., Sharma Dey, R., Bag, P., Chakraborti, S., & Khan, M. T. H. (2009). Recent advancements for the evaluation of anti-viral activities of natural products. New Biotechnology, 25(5), 347-368. doi:10.1016/j.nbt.2009.03.00

Characterization and frequency of a newly identified HIV-1 BF1 intersubtype circulating recombinant form in São Paulo, Brazil

Background: HIV circulating recombinant forms (CRFs) play an important role in the global and regional HIV epidemics, particularly in regions where multiple subtypes are circulating. To date, several (>40) CRFs are recognized worldwide with five currently circulating in Brazil. Here, we report the characterization of near full-length genome sequences (NFLG) of six phylogenetically related HIV-1 BF1 intersubtype recombinants (five from this study and one from other published sequences) representing CRF46_BF1.Methods: Initially, we selected 36 samples from 888 adult patients residing in São Paulo who had previously been diagnosed as being infected with subclade F1 based on pol subgenomic fragment sequencing. Proviral DNA integrated in peripheral blood mononuclear cells (PBMC) was amplified from the purified genomic DNA of all 36-blood samples by five overlapping PCR fragments followed by direct sequencing. Sequence data were obtained from the five fragments that showed identical genomic structure and phylogenetic trees were constructed and compared with previously published sequences. Genuine subclade F1 sequences and any other sequences that exhibited unique mosaic structures were omitted from further analysisResults: of the 36 samples analyzed, only six sequences, inferred from the pol region as subclade F1, displayed BF1 identical mosaic genomes with a single intersubtype breakpoint identified at the nef-U3 overlap (HXB2 position 9347-9365; LTR region). Five of these isolates formed a rigid cluster in phylogentic trees from different subclade F1 fragment regions, which we can now designate as CRF46_BF1. According to our estimate, the new CRF accounts for 0.56% of the HIV-1 circulating strains in São Paulo. Comparison with previously published sequences revealed an additional five isolates that share an identical mosaic structure with those reported in our study. Despite sharing a similar recombinant structure, only one sequence appeared to originate from the same CRF46_BF1 ancestor.Conclusion: We identified a new circulating recombinant form with a single intersubtype breakpoint identified at the nef-LTR U3 overlap and designated CRF46_BF1. Given the biological importance of the LTR U3 region, intersubtype recombination in this region could play an important role in HIV evolution with critical consequences for the development of efficient genetic vaccines.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Hemoctr, Fundacao Prosangue, São Paulo, BrazilUniversidade Federal de São Paulo, Retrovirol Lab, São Paulo, BrazilUniversidade Federal de São Paulo, Retrovirol Lab, São Paulo, BrazilFAPESP: 06/50096-0FAPESP: 2004/15856-9FAPESP: 2007/04890-0Web of Scienc

Prevalência, fatores de risco e caracterização genética dos vírus linfotrópico de células T humana tipo 1 e 2 em pacientes infectados pelo vírus da imunodeficiência humana tipo 1 nas Cidades de Ribeirão Preto e São Paulo

The aim of this study was to define the prevalence of human T cell lymphotropic virus types 1 and 2 in patients who were positive for human immunodeficiency virus type 1 in the State of Sao Paulo, Brazil. We evaluated 319 individuals infected with HIV type 1 who were attended at specialized clinics in two cities (Ribeirao Preto and Sao Paulo). The patients were interviewed and tested for antibodies against HTLV types 1 and 2 (Ortho HTLV-1/HTLV-2 Ab-Capture enzyme immunoassay). Direct DNA sequencing of polymerase chain reaction products from the tax region of HTLV type 2 and the long terminal repeat region of HTLV types 1 and 2 were performed to differentiate and determine the subtypes. The overall prevalence of anti-HTLV type 1 and 2 antibodies was 7.5% (24/319; 95% CI: 5.2-11.5). HTLV type 1 and 2 infection was associated with a history of injected drug use and with antibodies for hepatitis C virus (p 0.05). HTLV DNA was detected in 13 out of 24 samples, of which 12 were characterized as HTLV subtype 2c and one as HTLV subtype 1a. Among the 12 HTLV type 2 samples, seven were from injected drug users, thus indicating that this route is an important risk factor for HTLV type 2 transmission among our population infected with HIV type 1.Fundacao Pr6 Sangue, Hemoctr Sao Paulo, BR-05403000 Sao Paulo, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Lab Retrovirol, Dept Doencas Infecciosas & Parasitarias, Sao Paulo, BrazilSecretaria Estado Saude Sao Paulo, Ctr Referencia & Treinamento DST Aids, Sao Paulo, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Lab Retrovirol, Dept Doencas Infecciosas & Parasitarias, Sao Paulo, BrazilWeb of Scienc

HIV-1 subtypes among intravenous drug users from two neighboring cities in São Paulo State, Brazil

In order to assess the molecular epidemiology of HIV-1 in two neighboring cities located near the epicenter of the HIV-1 epidemics in Brazil (Santos and São Paulo), we investigated 83 HIV-1 strains obtained from samples collected in 1995 from intravenous drug users. The V3 through V5 region of the envelope of gp 120 was analyzed by heteroduplex mobility analysis. Of the 95 samples, 12 (12.6%) were PCR negative (6 samples from each group); low DNA concentration was the reason for non-amplification in half of these cases. Of the 42 typed cases from São Paulo, 34 (81%, 95% confidence limits 74.9 to 87.0%) were B and 8 (19%, 95% confidence limits 12.9 to 25.0%) were F, whereas of the 41 typed cases from Santos, 39 (95%, 95% confidence limits 91.6 to 98.4%) were B and 2 (5%, 95% confidence limits 1.6 to 8.4%) were C. We therefore confirm the relationship between clade F and intravenous drug use in São Paulo, and the presence of clade C in Santos. The fact that different genetic subtypes of HIV-1 are co-circulating indicates a need for continuous surveillance for these subtypes as well as for recombinant viruses in Brazil.Instituto Adolfo LutzUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Laboratório de RetrovirologiaFundação Pró-Sangue/Hemocentro de São PauloCentro de Referência de AIDS (CRAIDS)UNIFESP, EPM, Laboratório de RetrovirologiaSciEL

An alternative storage method for characterization of the intestinal microbiota through next generation sequencing

Gut microbiota has been the subject of various molecular studies mainly due to its importance and wide-ranging relationships with human hosts. However, the storage of fecal samples prior to DNA extraction is critical when characterizing the composition of intestinal microbiota. Therefore, we aimed to understand the effects of different fecal storage methods to characterize intestinal microbiota using Next Generation Sequencing (NGS) as well as to establish an alternative conservation method of bacterial genetic material in these samples using guanidine. Stool samples from 10 healthy volunteers were collected. Each sample was divided into five aliquots: one aliquot was extracted immediately after collection (fresh) and two aliquots were subjected to freezing at -20 °C or -80 °C and extracted after 48 h. The other two aliquots were stored in guanidine at room temperature or 4 °C and extracted after 48 h. The V4 hypervariable regions of the bacterial and archeal 16S rRNA gene were amplified by PCR and sequenced using an Ion Torrent PGM platform for NGS. The data were analyzed using QIIME software. Statistical significance was determined using a nonparametric Kruskal-Wallis test. A total of 11,494,688 reads with acceptable quality were obtained. Unweighted principal coordinate analysis (PCoA) revealed that the samples were clustered based on the host rather than by the storage group. At the phylum and genus levels, we observed statistically significant differences between two genera, Proteobacteria (p=0.013) and Suterella (p=0.004), comparing frozen samples with guanidine-stored samples. Our data suggest that the use of guanidine can preserve bacterial genetic materials as well as freezing, providing additional convenience