JOAO RENATO REBELLO PINHO

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Instituto Central, Hospital das Clínicas, Faculdade de Medicina - Médico
LIM/07 - Laboratório de Gastroenterologia Clínica e Experimental, Hospital das Clínicas, Faculdade de Medicina
LIM/03 - Laboratório de Medicina Laboratorial, Hospital das Clínicas, Faculdade de Medicina

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Autor: CARRILHO, Flair J. ×

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  • article 16 Citação(ões) na Scopus
    Resistance mutations are rare among protease inhibitor treatment-naive hepatitis C genotype-1 patients with or without HIV coinfection
    (2015) LISBOA-NETO, Gaspar; NOBLE, Caroline F.; PINHO, Joao R. Rebello; MALTA, Fernanda M.; GOMES-GOUVEA, Michele S.; ALVARADO-MORA, Monica V.; SILVA, Mariliza H. da; LEITE, Andrea G. B.; PICCOLI, Leonora Z.; RODRIGUES, Flaviane K.; CARRILHO, Flair J.; MENDES-CORREA, Maria C.
    Background: HCV has a high replication rate and a lack of proofreading activity, leading to a greatly diverse viral population. This diversity may lead to emergence of resistant strains in direct-acting antiviral therapy. The frequency of naturally occurring HCV protease inhibitor (PI) mutations has been addressed in many countries, but there are few data on the prevalence of these mutations in Brazilian patients. Methods: We evaluated the sequence of HCV NS3 protease gene in 247 patients (135 HCV-monoinfected and 112 HIV-HCV-coinfected patients). HCV RNA was extracted from plasma and a fragment of 765 base pairs from the NS3 region was amplified and sequenced with Sanger-based technology. Results: HIV-HCV-coinfected patients were more likely to be older than 40 years and have an HCV subtype-1a infection. Overall, 21.9% of patients had at least one amino acid substitution in the NS3 region; 14 patients (5.7%) harboured at least one resistance mutation (T54S, V55A, Q80R) and the Q80K mutation was not found in our case series. There was no difference between monoinfected and coinfected patients regarding the frequency of natural polymorphisms and resistance mutations. Conclusions: Baseline HCV NS3 amino acid substitutions identified herein are considered mostly natural polymorphisms with no clinical impact on PI-based therapy. The identified resistance mutations may be associated with low-level resistance to PIs in vitro. Q80K substitution seems to be a rare event in Brazil. HIV coinfection was not associated with a greater frequency of such substitutions in the studied sample.
  • article 8 Citação(ões) na Scopus
    Distribution of hepatitis c virus (hcv) genotypes in patients with chronic infection from Rondonia, Brazil
    (2011) VIEIRA, Deusilene S.; ALVARADO-MORA, Monica V.; BOTELHO, Livia; CARRILHO, Flair J.; PINHO, Joao R. R.; SALCEDO, Juan M.
    Background: Hepatitis C virus (HCV) is an important human pathogen affecting around 3% of the human population. In Brazil, it is estimated that there are approximately 2 to 3 million HCV chronic carriers. There are few reports of HCV prevalence in Rondonia State (RO), but it was estimated in 9.7% from 1999 to 2005. The aim of this study was to characterize HCV genotypes in 58 chronic HCV infected patients from Porto Velho, Rondonia (RO), Brazil. Methods: A fragment of 380 bp of NS5B region was amplified by nested PCR for genotyping analysis. Viral sequences were characterized by phylogenetic analysis using reference sequences obtained from the GenBank (n = 173). Sequences were aligned using Muscle software and edited in the SE-AL software. Phylogenetic analyses were conducted using Bayesian Markov chain Monte Carlo simulation (MCMC) to obtain the MCC tree using BEAST v. 1.5.3. Results: From 58 anti-HCV positive samples, 22 were positive to the NS5B fragment and successfully sequenced. Genotype 1b was the most prevalent in this population (50%), followed by 1a (27.2%), 2b (13.6%) and 3a (9.0%). Conclusions: This study is the first report of HCV genotypes from Rondonia State and subtype 1b was found to be the most prevalent. This subtype is mostly found among people who have a previous history of blood transfusion but more detailed studies with a larger number of patients are necessary to understand the HCV dynamics in the population of Rondonia State, Brazil.
  • conferenceObject
    Serum Lipidomic Profiling for Screening Potential Biomarkers of Liver Cirrhosis among Patients with Chronic Hepatitis C
    (2015) PASSOS-CASTILHO, Ana Maria; FERRAZ, Maria Lucia; CARVALHO, Valdemir M.; CARDOZO, Karina H.; KIKUCHI, Luciana; CHAGAS, Aline; PINHO, Joao Renato R.; GOMES-GOUVEA, Michele S.; MALTA, Fernanda; CARRILHO, Flair J.; GRANATO, Celso
  • article 4 Citação(ões) na Scopus
    The Molecular Characterization of Hepatitis A Virus Strains Circulating during Hepatitis A Outbreaks in Sao Paulo, Brazil, from September 2017 to May 2019
    (2022) CHUFFI, Samira; GOMES-GOUVEA, Michele S.; CASADIO, Luciana V. B.; NASTRI, Ana Catharina S. S.; GONZALEZ, Mario P.; COTIA, Andre L. F.; ARANDA, Amanda G. D.; TENORE, Simone B.; ONO, Suzane K.; MALTA, Fernanda M.; MADALOSSO, Geraldine; FERREIRA, Paulo R. A.; CARRILHO, Flair J.; PINHO, Joao R. R.
    Outbreaks of hepatitis A may occur in countries of medium and high socioeconomic levels in which the population generally exhibits an increased susceptibility in young adults to this infection if they are not vaccinated against the hepatitis A virus (HAV). In Europe, an outbreak involved approximately 22 European countries with 4475 cases reported from 2016 to 2018; most of them were men who have sex with men (MSM). This outbreak expanded to North and South America, including Brazil, particularly in Sao Paulo city with 1547 reported cases from 2016 to 2019. In the present study, we characterized the HAV strains involved in the acute hepatitis A cases identified in the reference centers of Sao Paulo city during this outbreak. A total of 51 cases with positive anti-HAV IgM were included, 80.4% male, 68.6% of them between 20 and 40 years old and 41.7% MSM. HAV RNA was detected in 92% (47/51) of the cases. Subgenotype IA of HAV was identified and most of the strains were closely related to that isolated in outbreaks that occurred in different European countries in 2016. These results showed the epidemiological relation between these outbreaks and reinforce the need to implement vaccination against hepatitis A for the adult population, particularly for a population with a high-risk behavior.
  • article 13 Citação(ões) na Scopus
    Phylogenetic analysis of complete genome sequences of hepatitis B virus from an Afro-Colombian community: presence of HBV F3/A1 recombinant strain
    (2012) ALVARADO-MORA, Monica V.; ROMANO, Camila M.; GOMES-GOUVEA, Michele S.; GUTIERREZ, Maria F.; CARRILHO, Flair J.; PINHO, Joao R. R.
    Background: Hepatitis B virus (HBV) infection is one of the most prevalent viral infections in humans and represents a serious public health problem. In Colombia, our group reported recently the presence of subgenotypes F3, A2 and genotype G in Bogota. The aim of this study was to characterize the HBV genotypes circulating in Quibdo, the largest Afro-descendant community in Colombia. Sixty HBsAg-positive samples were studied. A fragment of 1306 bp (S/POL) was amplified by nested PCR. Positive samples to S/POL fragment were submitted to PCR amplification of the HBV complete genome. Findings: The distribution of HBV genotypes was: A1 (52.17%), E (39.13%), D3 (4.3%) and F3/A1 (4.3%). An HBV recombinant strain subgenotype F3/A1 was found for the first time. Conclusions: This study is the first analysis of complete HBV genome sequences from Afro-Colombian population. It was found an important presence of HBV/A1 and HBV/E genotypes. A new recombinant strain of HBV genotype F3/A1 was reported in this population. This fact may be correlated with the introduction of these genotypes in the times of slavery.
  • article 8 Citação(ões) na Scopus
    High Prevalence of Hepatitis B Subgenotype D4 in Northeast Brazil: an Ancient Relic from African Continent?
    (2018) CRUZ-SANTOS, Max D.; GOMES-GOUVEA, Michele S.; COSTA-NUNES, Jomar D.; MALTA-ROMANO, Camila; TELES-SOUSA, Marinilde; FONSECA-BARROS, Lena M.; CARRILHO, Flair J.; PAIVA-FERREIRA, Adalgisa de S.; REBELLO-PINHO, Joao R.
    Introduction. Hepatitis B virus (HBV) infection leads to a chronic liver disease that is distributed worldwide. The characterization of HBV into genotypes/subgenotypes is not only a mere procedure for distinguishing different HBV strains around the world because determining their geographic distribution is crucial to understanding their spread across the world. Material and methods. We characterized different HBV genotypes and subgenotypes in five municipalities located in northeastern Maranhao, in the Brazilian north Atlantic coast. 92 HBsAg-positive individuals were submitted to PCR (polymerase chain reaction). Fifty samples were sequenced using automated Sanger sequencing and classified by phylogenetic methods. Results. Subgenotypes D4 and A1 were found in 42 (84%) and eight (16%) samples, respectively. To our knowledge, this is the first study to describe a high frequency of subgenotype D4 in any population. Subgenotype A1 is frequently found across Brazil, but D4 has been rarely detected and only in a few Brazilian states. This study shows the characterization of HBV subgenotypes from a population based study in the state of Maranhao, particularly in populations that do not have frequent contact with populations from other regions of the world. Conclusion. Our findings showed a HBV subgenotype profile that probably reflect the viruses that were brought with the slave trade from Africa to Maranhao. This study also reinforces the need to evaluate the status of HBV dispersion not only in large urban centers, but also in the hinterland, to enable the implementation of effective control and treatment measures.
  • article 6 Citação(ões) na Scopus
    The role of PNPLA3 and TM6SF2 polymorphisms on liver fibrosis and metabolic abnormalities in Brazilian patients with chronic hepatitis C
    (2021) OLIVEIRA, Arthur Ivan N.; MALTA, Fernanda M.; ZITELLI, Patricia Momoyo Y.; SALLES, Ana Paula M.; GOMES-GOUVEA, Michele S.; NASTRI, Ana Catharina S.; PINHO, Joao Renato R.; CARRILHO, Flair J.; OLIVEIRA, Claudia P.; MENDES-CORREA, Maria Cassia; PESSOA, Mario G.; MAZO, Daniel F.
    BackgroundDespite the growing body of knowledge about TM6SF2 and PNPLA3 polymorphisms in non-alcoholic fatty liver disease, their influence in the spectrum of HCV liver disease is not yet fully defined. Besides that, admixed populations, such as Brazilians, were not included in most of the studies.MethodsThis cross-sectional study enrolled 365 treatment-naive patients with HCV and 134 healthy individuals. TM6SF2 (rs58542926 c.499C>T) and PNPLA3 (rs738409 c.444C>G) polymorphisms were evaluated regarding their association with clinical and laboratory data, histological liver steatosis and fibrosis, and with components of the metabolic syndrome.ResultsIn HCV subjects, the frequencies of TM6SF2 CC and CT+TT were 89% and 11%, while PNPLA3 frequencies of CC and CG+GG were 51.4% and 48.6%. In the univariate logistic regression analysis, the TM6SF2 CT+TT genotype in HCV was associated with significant liver fibrosis (p=0.047; OR 1.953; 95% CI 1.009-3.788). In comparison to the CT+TT genotype, the TM6SF2 CC genotype in HCV was associated with older age (p=0.002), higher frequency of arterial hypertension (p=0.032), obesity (p=0.030), metabolic syndrome (p=0.014) and lower total cholesterol levels (p=0.036). The PNPLA3 GG subjects had lower body mass index than CG/ CC individuals (p=0.047). None of the polymorphisms, or their combinations, was independently associated with hepatic steatosis or fibrosis. On the other hand, older age, lower serum levels of total cholesterol, and higher serum levels of alanine aminotransferase and alkaline phosphatase were associated with liver fibrosis in the multivariate logistic regression analysis.ConclusionIn this evaluation of an admixed HCV population, neither TM6SF2 nor PNPLA3 polymorphisms were independently associated with hepatic steatosis or fibrosis. Other factors seem more influential than these specific polymorphisms in isolation. More studies are warranted to clarify the role of the TM6SF2 and PNPLA3 polymorphisms in Brazilians with HCV.
  • conferenceObject
    HBV resistance frequency in nucleos(t)ide analogue-untreated patients from different Brazilian regions
    (2012) GOMES-GOUVEA, Michele S.; MENDES-CORREA, Maria Cassia; FERREIRA, Ariana C.; TEIXEIRA, Rosangela; ANDRADE, Jose R.; BARROS, Lena Maria F.; FERREIRA, Adalgisa S.; REZENDE, Rosamar F.; NASTRI, Ana Catharina S.; LEITE, Andrea G.; PICCOLI, Leonora Z.; GALVAN, Josiane; CONDE, Simone Regina S.; SOARES, Manoel C.; CARRILHO, Flair J.; PINHO, Joao R.
    Background - Presence of viral variants with drug-resistance mutations in drug-naïve hepatitis B virus (HBV) infected patients can seriously compromise response to antiviral treatment. Therefore, our study aimed to determine the prevalence of HBV with drug-resistance in 557 untreated chronic hepatitis B (CHB) patients with nucleos(t)ide analogues (NA) from five different geographic regions of Brazil. Methods – HBV reverse-tran-scriptase (RT) region was sequenced and mutations associated with resistance to NA inhibitors were analyzed. Amino acid changes potentially associated with resistance were also investigated. Furthermore, HBV genotypes and subgenotypes were determined by phylogenetic analysis of the sequences. Results – HBV genotypes A [A1 (66.8%), A2 (2.3%)] and D [D1 (0.5%), D2 (4.3%), D3 (11.8%), D4 (6.6%)] were the most prevalent in Brazil, but genotypes B1 (0.2%), B2 (0.2%), C2 (0.7%), E (0.7%), F2a (4.5%), F4 (0.4%) and G (0.5%) were also found. Overall, 1.8% (10/557) of the patients carried HBV variants with primary drug-resistance mutations [rtM204V/I (0.4%); rtM204V + rtS202G (0.4%); rtA181S/T (0.4%); rtM250I (0.2%); rtA194T (0.4%)]. The four strains with mutation at position 204 also had compensatory mutations [rtL180M (3/4); rtL180M + rtV207I (1/4). One patient was infected with HBV variant only with compensatory mutations (rtV173L + rtL180M). Thirty (5%) patients were infected with strain harboring some of that mutation potentially associated with Adefovir resistance [rtS85A (n=1), rtV214A (n=6), rtQ215S (n=14), rtI233V (n=6), rtN238T (n=4), rtN238D (n=5)]. Additionally, we observed in 18 (3.2%) patients the presence of variants with novel amino acid substitutions at positions reported to be potentially associated with Adefovir resistance: rtV214G (n=2), V214E (n=1), Q215H (n=6), Q215P (n=1), E218D (n=2), I233L (n=1), T237A (n=1), N238H (n=3), N238A (n=1). Conclusions – HBV variability is high in Brazil, thirteen HBV subgenotypes were found. The rate of important drug resistance mutations was low (1.8%) among the drug-naïve HBV infected patients studied, indicating the high potential to full efficacy of nucleos(t)ide analogue therapy in these patients. The high frequency of mutations potentially associated with Adefovir resistance among untreated patients suggests that these mutations are not really associated to resistance.
  • article 14 Citação(ões) na Scopus
    Detection of Hepatitis B virus subgenotype A1 in a Quilombo community from Maranhao, Brazil
    (2011) ALVARADO-MORA, Monica V.; BOTELHO, Livia; GOMES-GOUVEA, Michele S.; SOUZA, Vanda F. de; NASCIMENTO, Maria C.; PANNUTI, Claudio S.; CARRILHO, Flair J.; PINHO, Joao R. R.
    Background: The Brazilian population is mainly descendant from European colonizers, Africans and Native Americans. Some Afro-descendants lived in small isolated communities since the slavery period. The epidemiological status of HBV infection in Quilombos communities from northeast of Brazil remains unknown. The aim of this study was to characterize the HBV genotypes circulating inside a Quilombo isolated community from Maranhao State, Brazil. Methods: Seventy-two samples from Frechal Quilombo community at Maranhao were collected. All serum samples were screened by enzyme-linked immunosorbent assays for the presence of hepatitis B surface antigen ( HBsAg). HBsAg positive samples were submitted to DNA extraction and a fragment of 1306 bp partially comprising HBsAg and polymerase coding regions (S/POL) was amplified by nested PCR and its nucleotide sequence was determined. Viral isolates were genotyped by phylogenetic analysis using reference sequences from each genotype obtained from GenBank (n = 320). Sequences were aligned using Muscle software and edited in the SE-AL software. Bayesian phylogenetic analyses were conducted using Markov Chain Monte Carlo (MCMC) method to obtain the MCC tree using BEAST v.1.5.3. Results: Of the 72 individuals, 9 (12.5%) were HBsAg-positive and 4 of them were successfully sequenced for the 1306 bp fragment. All these samples were genotype A1 and grouped together with other sequences reported from Brazil. Conclusions: The present study represents the first report on the HBV genotypes characterization of this community in the Maranhao state in Brazil where a high HBsAg frequency was found. In this study, we reported a high frequency of HBV infection and the exclusive presence of subgenotype A1 in an Afro-descendent community in the Maranhao State, Brazil.
  • article 16 Citação(ões) na Scopus
    Increased hepatic expression of miRNA-122 in patients infected with HCV genotype 3
    (2016) OLIVEIRA, Ketti G.; MALTA, Fernanda M.; NASTRI, Ana C. S. S.; WIDMAN, Azzo; FARIA, Paola L.; SANTANA, Rubia A. F.; ALVES, Venancio A. F.; CARRILHO, Flair J.; PINHO, Joao R. R.
    Hepatitis C virus (HCV) infection affects approximately 3 % of the world population. HCV targets hepatic tissue, and most infected patients develop a chronic infection. Currently, studies have demonstrated an association between HCV-RNA replication and miR-122, the most abundant microRNA in the liver. Our aim was to evaluate liver and serum expression of miR-122 in patients infected with HCV genotypes 1 and 3, and to identify possible associations between miR-122 expression and lipid profiles, HCV viral load, apolipoproteins and liver enzymes. MicroRNAs were isolated from blood and liver tissue, and miR-122 expression was quantified by real-time PCR. HCV viral load was quantified by real-time PCR and HCV genotype, and serum biomarkers were obtained from medical report. The levels of miR-122 were higher in liver than those in blood from individuals infected with HCV genotypes 1 and 3 (p < 0.0001). The tissue levels of miR-122 were higher in subjects infected with HCV genotype 3 (6.22-fold, p < 0.001). A positive correlation was observed between the blood and hepatic levels of miR-122 in patients infected with HCV genotype 1 (r = 0.302, p = 0.026); in these patients, an inverse correlation was observed between serum apolipoprotein A-II (ApoA-II) levels and the blood (r = -0.330; p = 0.014) and hepatic (r = -0.311; p = 0.020) levels of miR-122. In patients infected with HCV genotype 3, there was a positive correlation between the hepatic miR-122 and the high-density lipoprotein-HDL (r = 0.412, p = 0.036) and insulin (r = 0.478, p = 0.044). Lipid metabolism proteins and miR-122 expression levels have different relations in HCV-3- and HCV-1-infected patients.