109 results on '"Sampaio SC"'
Search Results
2. Effect of DHA-rich fish oil supplementation on human leukocyte function
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Curi, R., Gorjao, R., Verlengia, R., Soriano, Fg, Lima, Tm, Boaventura, Mfc, Kanunfre, C., Rosemari Otton, Peres, Cm, Sampaio, Sc, Martins, Ef, Folador, A., Curi, Tp, Lagranha, C., and Portiolli, E.
3. Unveiling viral pathogens in acute respiratory disease: Insights from viral metagenomics in patients from the State of Alagoas, Brazil.
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de Campos GM, Santos HO, Lima ARJ, Leite AB, Ribeiro G, Todão Bernardino JS, do Nascimento JPM, Souza JVC, de Lima LPO, Lima MBZ, de Araújo MA, Giovanetti M, Kallas EG, Sampaio SC, Elias MC, and Slavov SN
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Background: Respiratory illness affects individuals across all age demographics on a global scale, often precipitated by viral infections. The symptomatic manifestations of these diseases bear clinical resemblance, complicating the accurate determination of their etiological origins. Furthermore, the diagnostic panels for respiratory pathogens used within local medical practices, may not encompass the full spectrum of viral agents responsible for such ailments. Consequently, a significant number of clinically important viral pathogens may remain undetected., Methods and Findings: In the light of this, we conducted a metagenomic examination of 66 nasopharyngeal swab specimens, obtained from patients presenting with acute respiratory conditions yet tested negative by the standard diagnostic panels available locally. These specimens were obtained from the Public Health Laboratory, Maceio, State of Alagoas. Our findings indicate a predominant diagnostic escape of rhinoviruses and notably enterovirus D68. Moreover, our study identified a substantial quantity of sequence reads attributed to human respirovirus 3 (human parainfluenza 3) along with various herpresviruses including human herpesvirus-1, Epstein-Barr virus (Human herpesvirus-4), Human herpesviruses 6 and 7 and human parvovirus B19 (B19V). Notably, the metagenomic analysis uncovered a widespread presence of the emerging human vientovirus FB in most of sample pools, though its clinical importance remains to be elucidated., Conclusions: The obtained results in this study underscore the invaluable role of viral metagenomics in the identification of underrecognized viruses bearing clinical relevance. Furthermore, it offers insights into the dissemination of these pathogens within the studied area, thereby informing public health strategies aimed at enhancing diagnostic accuracy and improving patient care., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 de Campos et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2024
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4. A new lineage nomenclature to aid genomic surveillance of dengue virus.
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Hill V, Cleemput S, Pereira JS, Gifford RJ, Fonseca V, Tegally H, Brito AF, Ribeiro G, de Souza VC, Brcko IC, Ribeiro IS, De Lima ITT, Slavov SN, Sampaio SC, Elias MC, Tran VT, Kien DTH, Huynh T, Yacoub S, Dieng I, Salvato R, Wallau GL, Gregianini TS, Godinho FMS, Vogels CBF, Breban MI, Leguia M, Jagtap S, Roy R, Hapuarachchi C, Mwanyika G, Giovanetti M, Alcantara LCJ, Faria NR, Carrington CVF, Hanley KA, Holmes EC, Dumon W, Lima ARJ, Oliveira T, and Grubaugh ND
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- Humans, Genotype, Genomics methods, Genetic Variation, Terminology as Topic, Dengue Virus genetics, Dengue Virus classification, Phylogeny, Genome, Viral, Dengue virology, Dengue epidemiology
- Abstract
Dengue virus (DENV) is currently causing epidemics of unprecedented scope in endemic settings and expanding to new geographical areas. It is therefore critical to track this virus using genomic surveillance. However, the complex patterns of viral genomic diversity make it challenging to use the existing genotype classification system. Here, we propose adding 2 sub-genotypic levels of virus classification, named major and minor lineages. These lineages have high thresholds for phylogenetic distance and clade size, rendering them stable between phylogenetic studies. We present assignment tools to show that the proposed lineages are useful for regional, national, and subnational discussions of relevant DENV diversity. Moreover, the proposed lineages are robust to classification using partial genome sequences. We provide a standardized neutral descriptor of DENV diversity with which we can identify and track lineages of potential epidemiological and/or clinical importance. Information about our lineage system, including methods to assign lineages to sequence data and propose new lineages, can be found at: dengue-lineages.org., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: SC and WD are affiliated with emweb. NDG is a paid consultant for BioNTech., (Copyright: © 2024 Hill et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2024
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5. Unveiling microbial worlds: exploring viral metagenomics among waste pickers at Latin America's largest dumpsite.
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Cruvinel VRN, Carvalho E, Alves DCC, Marques CP, Bezerra RDS, Giovanetti M, Sampaio SC, Elias MC, Araújo WN, Haddad R, and Slavov SN
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- Humans, Brazil, Male, Adult, Female, Viruses genetics, Viruses classification, Viruses isolation & purification, Occupational Exposure, Case-Control Studies, Middle Aged, Refuse Disposal, Metagenomics
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Waste pickers constitute a marginalized demographic engaged in the collection of refuse, facing considerable occupational hazards that heighten their susceptibility to contract infectious diseases. Moreover, waste pickers contend with societal stigmatization and encounter barriers to accessing healthcare services. To explore the viral profile of waste pickers potentially linked to their occupational environment, we conducted a metagenomic analysis on 120 plasma specimens sampled from individuals employed at the Cidade Estrutural dumpsite in Brasilia city, Brazil. In total, 60 blood donors served as a comparative control group. Specimens were pooled and subjected to Illumina NextSeq 2000 sequencing. Viral abundance among waste pickers revealed the presence of significant pathogens, including HIV, HCV, and Chikungunya, which were not detected in the control group. Additionally, elevated levels of anelloviruses and Human pegivirus-1 were noted, with a comparable incidence in the control group. These findings underscore the utility of metagenomics in identifying clinically relevant viral agents within underserved populations. The implications of this study extend to informing public health policies aimed at surveilling infectious diseases among individuals facing socioeconomic disparities and limited access to healthcare resources.
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- 2024
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6. Dynamic clade transitions and the influence of vaccination on the spatiotemporal circulation of SARS-CoV-2 variants.
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Banho CA, de Carvalho Marques B, Sacchetto L, Lima AKS, Parra MCP, Lima ARJ, Ribeiro G, Martins AJ, Barros CRDS, Elias MC, Sampaio SC, Slavov SN, Rodrigues ES, Santos EV, Covas DT, Kashima S, Brassaloti RA, Petry B, Clemente LG, Coutinho LL, Assato PA, da Silva da Costa FA, Grotto RMT, Poleti MD, Lesbon JCC, Mattos EC, Fukumasu H, Giovanetti M, Alcantara LCJ, Souza-Neto JA, Rahal P, Araújo JP Jr, Spilki FR, Althouse BM, Vasilakis N, and Nogueira ML
- Abstract
Since 2021, the emergence of variants of concern (VOC) has led Brazil to experience record numbers of in COVID-19 cases and deaths. The expanded spread of the SARS-CoV-2 combined with a low vaccination rate has contributed to the emergence of new mutations that may enhance viral fitness, leading to the persistence of the disease. Due to limitations in the real-time genomic monitoring of new variants in some Brazilian states, we aimed to investigate whether genomic surveillance, coupled with epidemiological data and SARS-CoV-2 variants spatiotemporal spread in a smaller region, can reflect the pandemic progression at a national level. Our findings revealed three SARS-CoV-2 variant replacements from 2021 to early 2022, corresponding to the introduction and increase in the frequency of Gamma, Delta, and Omicron variants, as indicated by peaks of the Effective Reproductive Number (Reff). These distinct clade replacements triggered two waves of COVID-19 cases, influenced by the increasing vaccine uptake over time. Our results indicated that the effectiveness of vaccination in preventing new cases during the Delta and Omicron circulations was six and eleven times higher, respectively, than during the period when Gamma was predominant, and it was highly efficient in reducing the number of deaths. Furthermore, we demonstrated that genomic monitoring at a local level can reflect the national trends in the spread and evolution of SARS-CoV-2., (© 2024. The Author(s).)
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- 2024
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7. Exploring the Chikungunya virus landscape in a dengue-endemic Brazilian area.
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de La Roque DGL, Santos EV, Policastro LR, da Costa PNM, Evaristo M, Yamamoto AY, Giomo DB, Torres PMA, Gentil DCD, Minto ECM, Slavov SN, Fonseca V, Dos Santos Barros CR, Martins AJ, Calado RT, Passos LMR, Elias MC, Sampaio SC, Giovanetti M, Covas DT, Alcântara LCJ, and Kashima S
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- Humans, Brazil epidemiology, Male, Female, Adult, Middle Aged, Young Adult, Endemic Diseases, Adolescent, Whole Genome Sequencing, Aged, Child, Phylogeny, Mutation, Child, Preschool, Dengue Virus genetics, Dengue Virus isolation & purification, Dengue Virus classification, Thrombocytopenia epidemiology, Thrombocytopenia virology, Chikungunya Fever epidemiology, Chikungunya Fever blood, Chikungunya Fever virology, Chikungunya virus genetics, Chikungunya virus isolation & purification, Dengue epidemiology, Dengue virology, Genotype, RNA, Viral genetics
- Abstract
We aimed to describe the landscape, including molecular, epidemiological, and clinical aspects of CHIKV infections in the Ribeirao Preto region, an area endemic to dengue. We randomly screened 3744 plasma samples that had undergone DENV diagnosis to evaluate CHIKV-RNA using an in-house RT-PCR assay. Positive samples were followed clinically, and RNA samples were submitted to whole genome sequencing. Seventeen cases (0.5 %) were positive for CHIKV-RNA despite being negative for DENV-RNA. Notably, half of the patients experienced prolonged arthralgia lasting more than 90 days. Compared with the healthy control group, leukopenia and thrombocytopenia were observed in all CHIKV-positive individuals with statistically significant P values (P < 0.0001 and P = 0.0003, respectively). The genomic analysis revealed that the CHIKV strains being studied are classified within the East-Central-South-African (ECSA) genotype. This analysis identified new mutations, E1: K211E and E2: V264A, while the previously known mutation E1: A226V was not detected among these strains. This study highlights the need for epidemiological surveillance and preparedness for potential CHIKV epidemics in Brazil, particularly where other arboviruses co-circulate., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)
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- 2024
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8. Correction: Bufalo et al. Human Sensory Neuron-like Cells and Glycated Collagen Matrix as a Model for the Screening of Analgesic Compounds. Cells 2022, 11 , 247.
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Bufalo MC, Almeida MES, Jensen JR, DeOcesano-Pereira C, Lichtenstein F, Picolo G, Chudzinski-Tavassi AM, Sampaio SC, Cury Y, and Zambelli VO
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In the original publication [...].
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- 2024
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9. Simultaneous Dengue and Chikungunya Coinfection in Endemic Area in Brazil: Clinical Presentation and Implications for Public Health.
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Torres PMA, Roque DGL, Policastro LR, Chagas LBMO, Giomo DB, Gentil DCD, Fonseca V, Elias MC, Sampaio SC, Giovanetti M, Fonseca BAL, Calado RT, Alcantara LC, Covas DT, Santos FLS, Kashima S, and Passos LMR
- Abstract
Background: Dengue virus (DENV) and Chikungunya virus (CHIKV) pose significant public health threats in Brazil, where favorable conditions facilitated the proliferation of Aedes mosquitoes. Since the mid-1980s, Brazil has experienced annual outbreaks of DENV, with recent increases in confirmed cases. In addition, CHIKV, which was first reported in 2014, has spread across the country. The concurrent presence of these viruses has triggered public health alerts in endemic regions, underscoring the complexity of managing vector-borne diseases., Case Presentation: This report details a case of simultaneous DENV and CHIKV infections. A 77-year-old female patient who has diabetes and arrhythmia exhibited symptoms including fever, myalgia, and severe arthralgia. Laboratory tests confirmed the coinfection through RNA detection. The patient received supportive care, showed gradual improvement, and was eventually discharged., Conclusions: Coinfection with DENV and CHIKV cases reported here developed with mild outcomes. However, one of the patients did not recover from the arthralgia after presenting diagnostic challenges, which underscores the need for accurate differentiation to manage symptoms effectively. The reported cases, amidst increasing DENV outbreaks, highlight the urgency for preparedness in the healthcare system. The Ribeirão Preto region's endemicity for DENV, coupled with the rising incidence of CHIKV, emphasizes the evolving landscape of arbovirus transmission. Studies on Aedes mosquitoes suggest potential implications for human infection dynamics, warranting further investigation into arbovirus transmission efficacy and coinfection dynamics., Competing Interests: Competing interests The authors declare that they have no financial and non-financial competing interests.
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- 2024
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10. Metagenomic Analysis for Diagnosis of Hemorrhagic Fever in Minas Gerais, Brazil.
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Iani FCM, de Campos GM, Adelino TER, da Silva AS, Kashima S, Alcantara LCJ, Sampaio SC, Giovanetti M, Elias MC, and Slavov SN
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Viral hemorrhagic fever poses a significant public health challenge due to its severe clinical presentation and high mortality rate. The diagnostic process is hindered by similarity of symptoms across different diseases and the broad spectrum of pathogens that can cause hemorrhagic fever. In this study, we applied viral metagenomic analysis to 43 serum samples collected by the Public Health Laboratory ( Fundação Ezequiel Dias , FUNED) in Minas Gerais State, Brazil, from patients diagnosed with hemorrhagic fever who had tested negative for the standard local hemorrhagic disease testing panel. This panel includes tests for Dengue virus (DENV) IgM, Zika virus IgM, Chikungunya virus IgM, yellow fever IgM, Hantavirus IgM, Rickettsia rickettsii IgM/IgG, and Leptospira interrogans IgM, in addition to respective molecular tests for these infectious agents. The samples were grouped into 18 pools according to geographic origin and analyzed through next-generation sequencing on the NextSeq 2000 platform. Bioinformatic analysis revealed a prevalent occurrence of commensal viruses across all pools, but, notably, a significant number of reads corresponding to the DENV serotype 2 were identified in one specific pool. Further verification via real-time PCR confirmed the presence of DENV-2 RNA in an index case involving an oncology patient with hemorrhagic fever who had initially tested negative for anti-DENV IgM antibodies, thereby excluding this sample from initial molecular testing. The complete DENV-2 genome isolated from this patient was taxonomically classified within the cosmopolitan genotype that was recently introduced into Brazil. These findings highlight the critical role of considering the patient's clinical condition when deciding upon the most appropriate testing procedures. Additionally, this study showcases the potential of viral metagenomics in pinpointing the viral agents behind hemorrhagic diseases. Future research is needed to assess the practicality of incorporating metagenomics into standard viral diagnostic protocols.
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- 2024
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11. DENV-1 genotype V circulation during the nonepidemic period in the Northeast of São Paulo State endemic area.
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de La-Roque DGL, Santos EV, Oliveira RAM, Slavov SN, Rodrigues ES, Fonseca V, Martins AJ, Giomo DB, Torres PMA, Gentil DCD, Catoia EA, Chiquito NDC, Medeiros APSS, Yamamoto AY, Passos LMR, Calado RT, Barros CRDS, Elias MC, Sampaio SC, Giovanetti M, Junior Alcantara LC, Covas DT, and Kashima S
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- Humans, Brazil epidemiology, Genotype, Dengue Virus genetics, Dengue epidemiology
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- 2024
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12. Epidemiology of the SARS-CoV-2 Omicron Variant Emergence in the Southeast Brazilian Population.
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Rodrigues ES, Slavov SN, de La Roque DGL, Santos EV, Borges JS, Evaristo M, da Costa PNM, de Matos Maçonetto J, Marques AA, Baccarin AD, Oliveira RAM, Junior WL, Benincasa BI, de Andrade da Cruz LM, Lima ARJ, Ribeiro G, Viala VL, de Lima LPO, Martins AJ, Dos Santos Barros CR, Marqueze EC, de Souza Todao Bernardino J, Grotto RMT, Souza-Neto JA, Fonseca V, Nogueira ML, Fukumasu H, Coutinho LL, Calado RT, Covas DT, Giovanetti M, Alcantara LCJ, Sampaio SC, Elias MC, and Kashima S
- Abstract
The aim of this study was to describe epidemiological characteristics and perform SARS-CoV-2 genomic surveillance in the southeastern region of São Paulo State. During the first months of 2022, we compared weekly SARS-CoV-2 infection prevalence considering age, Ct value, and variants' lineages. An increase in the number of SARS-CoV-2-positive cases until the fourth epidemiological week of 2022 was observed. From the fourth epidemiological week onwards, the number of tests for SARS-CoV-2 diagnosis began to decrease, but the number of positive samples for SARS-CoV-2 remained high, reaching its most expressive level with a rate of 60% of infected individual cases. In this period, we observed a progressive increase in SARS-CoV-2 infection within the 0-10 age group throughout the epidemiological weeks, from 2.8% in the first epidemiological week to 9.2% in the eighth epidemiological week of 2022. We further observed significantly higher Ct values within younger patient samples compared to other older age groups. According to lineage assignment, SARS-CoV-2 (BA.1) was the most prevalent (74.5%) in the younger group, followed by BA.1.1 (23%), BA.2 (1.7%), and Delta (1%). Phylogenetic analysis showed that BA.2 sequences clustered together, indicating sustained transmission of this Omicron VOC sub-lineage by that time. Our results suggest the initial dissemination steps of the Omicron's sub-linage BA.2 into the younger group, due to specific genomic features of the detected sequences. These data provide interesting results related to the spread, emergence, and evolution of the Omicron variant in the southeast Brazilian population.
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- 2024
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13. Updated Insights into the Phylogenetics, Phylodynamics, and Genetic Diversity of Nipah Virus (NiV).
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de Campos GM, Cella E, Kashima S, Alcântara LCJ, Sampaio SC, Elias MC, Giovanetti M, and Slavov SN
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- Animals, Humans, Phylogeny, Bayes Theorem, Genetic Variation, Nipah Virus genetics, Chiroptera, Henipavirus Infections
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Nipah virus (NiV), a biosafety level 4 agent, was first identified in human clinical cases during an outbreak in 1998 in Malaysia and Singapore. While flying foxes are the primary host and viral vector, the infection is associated with a severe clinical presentation in humans, resulting in a high mortality rate. Therefore, NiV is considered a virus with an elevated epidemic potential which is further underscored by its recent emergence (September 2023) as an outbreak in India. Given the situation, it is paramount to understand the molecular dynamics of the virus to shed more light on its evolution and prevent potential future outbreaks. In this study, we conducted Bayesian phylogenetic analysis on all available NiV complete genomes, including partial N-gene NiV sequences (≥1000 bp) in public databases since the first human case, registered in 1998. We observed the distribution of genomes into three main clades corresponding to the genotypes Malaysia, Bangladesh and India, with the Malaysian clade being the oldest in evolutionary terms. The Bayesian skyline plot showed a recent increase in the viral population size since 2019. Protein analysis showed the presence of specific protein families (Hendra_C) in bats that might keep the infection in an asymptomatic state in bats, which also serve as viral vectors. Our results further indicate a shortage of complete NiV genomes, which would be instrumental in gaining a better understanding of NiV's molecular evolution and preventing future outbreaks. Our investigation also underscores the critical need to strengthen genomic surveillance based on complete NiV genomes that will aid thorough genetic characterization of the circulating NiV strains and the phylogenetic relationships between the henipaviruses. This approach will better prepare us to tackle the challenges posed by the NiV virus and other emerging viruses.
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- 2024
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14. Exploring Viral Metagenomics in Pediatric Patients with Acute Respiratory Infections: Unveiling Pathogens beyond SARS-CoV-2.
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de Campos GM, de La-Roque DGL, Lima ARJ, Zucherato VS, de Carvalho E, de Lima LPO, de Queiroz Cattony Neto P, Dos Santos MM, Ciccozzi M, Giovanetti M, Haddad R, Alcantara LCJ, Elias MC, Sampaio SC, Covas DT, Kashima S, and Slavov SN
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The emergence of SARS-CoV-2 and the subsequent pandemic have prompted extensive diagnostic and clinical efforts to mitigate viral spread. However, these strategies have largely overlooked the presence of other respiratory viruses. Acute respiratory diseases in pediatric patients can be caused by a diverse range of viral agents, and metagenomics represents a powerful tool for their characterization. This study aimed to investigate the viral abundance in pediatric patients with acute respiratory symptoms who tested negative for SARS-CoV-2 during the Omicron pandemic wave. To achieve this, viral metagenomics and next-generation sequencing were employed on 96 nasopharyngeal swab samples, which were organized into 12 pools, with each pool consisting of eight individual samples. Metagenomic analysis revealed that the most prevalent viruses associated with acute disease in pediatric patients were respiratory syncytial virus (detected in all pools) and enteroviruses, which are known to cause significant morbidity and mortality in children. Additionally, clinically significant viruses such as mumps orthorubulavirus, human metapneumovirus, influenza A, and a wide array of human herpesviruses (1, 3-7) were identified. These findings highlight the extensive potential of viral metagenomics in identifying viruses other than SARS-CoV-2 that contribute to acute infections in children. Consequently, this methodology should garner clinical attention in terms of differential diagnosis and the development of public policies to address such conditions in the global pediatric population.
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- 2023
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15. Epidemiological and Genomic Analysis of Asymptomatic SARS-CoV-2 Infections during the Delta and Omicron Epidemic Waves in São Paulo City, Brazil.
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Slavov SN, Lima ARJ, Ribeiro G, de Lima LPO, Barros CRDS, Marqueze EC, Martins AJ, Martininghi M, Palmieri M, Caldeira LAV, Silva FEVD, Cacherik G, Nicolodelli AL, Kashima S, Giovanetti M, Alcantara LCJ, Sampaio SC, and Elias MC
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- Humans, Brazil epidemiology, Asymptomatic Infections epidemiology, RNA, Viral genetics, SARS-CoV-2 genetics, Genomics, COVID-19 diagnosis, COVID-19 epidemiology
- Abstract
We examined the asymptomatic rates of SARS-CoV-2 infection during the Delta and Omicron waves in the city of São Paulo. Nasopharyngeal swabs were collected at strategic points of the city (open-air markets, bus terminals, airports) for SARS-CoV-2 RNA testing. Applying the questionnaire, the symptomatic individuals were excluded, and only asymptomatic cases were analyzed. During the Delta wave, a total of 4315 samples were collected, whereas 2372 samples were collected during the first Omicron wave. The incidence of the asymptomatic SARS-CoV-2 infection was 0.6% during the Delta wave and 0.8% during the Omicron wave. No statistical differences were found in the threshold amplification cycle. However, there was a statistical difference observed in the sublineage distribution between asymptomatic and symptomatic individuals. Our study determined the incidence of asymptomatic infection by monitoring individuals who remained symptom-free, thereby providing a reliable evaluation of asymptomatic SARS-CoV-2 carriage. Our findings reveal a relatively low proportion of asymptomatic cases, which could be attributed to our rigorous monitoring protocol for the presence of clinical symptoms. Investigating asymptomatic infection rates is crucial to develop and implement effective disease control strategies.
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- 2023
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16. Crotoxin Modulates Macrophage Phenotypic Reprogramming.
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Neves CL, Barbosa CMV, Ranéia-Silva PA, Faquim-Mauro EL, and Sampaio SC
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- Animals, Ascites, Macrophages, Macrophages, Peritoneal, Crotalus, Crotoxin pharmacology, Crotalid Venoms pharmacology
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Macrophage plasticity is a fundamental feature of the immune response since it favors the rapid and adequate change of the functional phenotype in response to the pathogen or the microenvironment. Several studies have shown that Crotoxin (CTX), the major toxin of the Crotalus durissus terrificus snake venom, has a long-lasting antitumor effect both in experimental models and in clinical trials. In this study, we show the CTX effect on the phenotypic reprogramming of macrophages in the mesenchymal tumor microenvironment or those obtained from the peritoneal cavity of healthy animals. CTX (0.9 or 5 μg/animal subcutaneously) administered concomitantly with intraperitoneal inoculation of tumor cells (1 × 10
7 /0.5 mL, injected intraperitoneally) of Ehrlich Ascitic Tumor (EAT) modulated the macrophages phenotype (M1), accompanied by increased NO• production by cells from ascites, and was evaluated after 13 days. On the other hand, in healthy animals, the phenotypic profile of macrophages was modulated in a dose-dependent way at 0.9 μg/animal: M1 and at 5.0 μg/animal: M2; this was accompanied by increased NO• production by peritoneal macrophages only for the dose of 0.9 μg/animal of CTX. This study shows that a single administration of CTX interferes with the phenotypic reprogramming of macrophages, as well as with the secretory state of cells from ascites, influencing events involved with mesenchymal tumor progression. These findings may favor the selection of new therapeutic targets to correct compromised immunity in different systems.- Published
- 2023
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17. Imidacloprid: Impact on Africanized Apis mellifera L. (Hymenoptera: Apidae) workers and honey contamination.
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Paloschi CL, Tavares MHF, Berte EA, Model K, Rosa KM, Conceição FGD, Domanski FR, de Souza Vismara E, Montanher PF, Maciel RMA, Ribeiro LDS, Ramos Mertz N, Sampaio SC, Costa FM, Lozano ER, and Potrich M
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- Bees, Animals, Ecosystem, Neonicotinoids toxicity, Nitro Compounds toxicity, Insecticides toxicity, Hymenoptera, Honey
- Abstract
Apis mellifera L. (Hymenoptera: Apidae) is fundamental in the production chain, ensuring food diversity through the ecosystem service of pollination. The aim of this work was to evaluate the impact of imidacloprid, orally, topically, and by contact, on A. mellifera workers and to verify the presence of this active ingredient in honey. Toxicity levels were verified by bioassays. In bioassay 1, the levels correspond to the percentages of 100, 10, 1, 0.1, and 0.01% of the recommended concentration for field application of the commercial product Nortox® (active ingredient imidacloprid), with which we obtained the mean lethal concentration (LC
50 ) in 48 h for A. mellifera, determining the concentration ranges to be used in the subsequent bioassays. Bioassays 2 and 3 followed the guidelines of the Organization for Economic Cooperation and Development, which specify the LC50 (48 h). In bioassay 4, the LC50 (48 h) and the survival rate of bees for a period of 120 h were determined by contact with a surface contaminated with imidacloprid, and in bioassay 5, the interference of the insecticide with the flight behavior of bees was evaluated. Honey samples were collected in agroecological and conventional georeferenced apiaries and traces of the imidacloprid were detected by means of high-performance liquid chromatography (HPLC-UV) with extraction by SPE C18. Bee survival was directly affected by the concentration and exposure time, as well behavioral performance, demonstrating the residual effect of imidacloprid on A. mellifera workers. Honey samples from a conventional apiary showed detection above the maximum residue limits (MRL) allowed by the European Union (0.05 μg mL-1 ), but samples from other apiaries showed no traces of this insecticide. Imidacloprid affects the survival rate and behavior of Africanized A. mellifera and honey quality., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Michele Potrich reports financial support was provided by National Council for Scientific and Technological Development. Cristiane Lurdes Paloschi reports financial support was provided by Coordination of Higher Education Personnel Improvement. Rodrigo Mendes Antunes Maciel reports financial support was provided by Coordination of Higher Education Personnel Improvement. Leticia da Silva Ribeiro reports was provided by National Council for Scientific and Technological Development., (Copyright © 2023 Elsevier Ltd. All rights reserved.)- Published
- 2023
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18. Human Pegivirus-1 Detection and Genotyping in Brazilian Patients with Fulminant Hepatitis.
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da Silva AS, de Campos GM, Villanova MG, Bezerra RDS, Santiago LMM, Haddad R, Covas DT, Giovanetti M, Alcantara LCJ, Elias MC, Sampaio SC, Kashima S, and Slavov SN
- Abstract
Fulminant hepatitis is a severe clinical disease characterized by a marked decline in liver function and encephalopathy. In a previous survey, using metagenomics in a group of 27 patients with this clinical condition, we observed an expressive quantity of reads of the Human pegivirus-1 (HPgV-1). Therefore, the objective of this study was to evaluate the frequency, molecular features, and HPgV-1 circulating genotypes in patients with fulminant hepatitis. After testing the collected plasma samples, we discovered twelve samples (44.4%) that were positive for HPgV-1 RNA (using both real-time and nested PCR). The positive samples presented a mean cycle threshold (Ct) of 28.5 (±7.3). Genotyping assignments revealed that all HPgV-1 positive samples belonged to the HPgV-1 genotype 2 (both subgenotypes 2A and 2B were identified). Although HPgV-1 is considered a commensal virus, little is known regarding its prevalence and genotypes in cases of fulminant hepatitis. More research is needed to understand whether HPgV-1 can be implicated in clinical disorders and infectious diseases.
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- 2023
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19. Retrospective Spatio-Temporal Dynamics of Dengue Virus 1, 2 and 4 in Paraguay.
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Vazquez C, Alcantara LCJ, Fonseca V, Lima M, Xavier J, Adelino T, Fritsch H, Castro E, de Oliveira C, Schuab G, Lima ARJ, Villalba S, Gomez de la Fuente A, Rojas A, Cantero C, Fleitas F, Aquino C, Ojeda A, Sequera G, Torales J, Barrios J, Elias MC, Iani FCM, Ortega MJ, Gamarra ML, Montoya R, Rodrigues ES, Kashima S, Sampaio SC, Coluchi N, Leite J, Gresh L, Franco L, Lourenço J, Rico JM, Bispo de Filippis AM, and Giovanetti M
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- Humans, Paraguay epidemiology, Retrospective Studies, Phylogeny, Serogroup, Genotype, Dengue Virus genetics, Dengue epidemiology
- Abstract
Dengue virus (DENV) has been a major public health concern in Paraguay, with frequent outbreaks occurring since early 1988. Although control measures have been implemented, dengue remains a significant health threat in the country, and continued efforts are required for prevention and control. In response to that, in collaboration with the Central Public Health Laboratory in Asunción, we conducted a portable whole-genome sequencing and phylodynamic analysis to investigate DENV viral strains circulating in Paraguay over the past epidemics. Our genomic surveillance activities revealed the co-circulation of multiple DENV serotypes: DENV-1 genotype V, the emerging DENV-2 genotype III, BR4-L2 clade, and DENV-4 genotype II. Results additionally highlight the possible role of Brazil as a source for the international dispersion of different viral strains to other countries in the Americas emphasizing the need for increased surveillance across the borders, for the early detection and response to outbreaks. This, in turn, emphasizes the critical role of genomic surveillance in monitoring and understanding arbovirus transmission and persistence locally and over long distances.
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- 2023
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20. Viral metagenomics unveils MW (Malawi) polyomavirus infection in Brazilian pediatric patients with acute respiratory disease.
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da Silva AS, de Campos GM, Giovanetti M, Zucherato VS, Lima ARJ, Santos EV, Haddad R, Ciccozzi M, Carlos Júnior Alcantara L, Elias MC, Sampaio SC, Covas DT, Kashima S, and Slavov SN
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- Infant, Child, Humans, Metagenomics, Brazil epidemiology, Malawi epidemiology, Phylogeny, SARS-CoV-2, COVID-19, Polyomavirus Infections epidemiology, Polyomavirus genetics, Viruses, Respiratory Tract Infections diagnosis, Respiratory Tract Infections epidemiology
- Abstract
Viral metagenomics has been extensively applied for the identification of emerging or poorly characterized viruses. In this study, we applied metagenomics for the identification of viral infections among pediatric patients with acute respiratory disease, but who tested negative for SARS-CoV-2. Twelve pools composed of eight nasopharyngeal specimens were submitted to viral metagenomics. Surprisingly, in two of the pools, we identified reads belonging to the poorly characterized Malawi polyomavirus (MWPyV). Then, the samples composing the positive pools were individually tested using quantitative polymerase chain reaction for identification of the MWPyV index cases. MWPyV-positive samples were also submitted to respiratory virus panel testing due to the metagenomic identification of different clinically important viruses. Of note, MWPyV-positive samples tested also positive for respiratory syncytial virus types A and B. In this study, we retrieved two complete MWPyV genome sequences from the index samples that were submitted to phylogenetic inference to investigate their viral origin. Our study represents the first molecular and genomic characterization of MWPyV obtained from pediatric patients in South America. The detection of MWPyV in acutely infected infants suggests that this virus might participate (coparticipate) in cases of respiratory symptoms. Nevertheless, future studies based on testing of a larger number of clinical samples and MWPyV complete genomes appear to be necessary to elucidate if this emerging polyomavirus might be clinically important., (© 2023 Wiley Periodicals LLC.)
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- 2023
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21. Global SARS-CoV-2 genomic surveillance: What we have learned (so far).
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Tosta S, Moreno K, Schuab G, Fonseca V, Segovia FMC, Kashima S, Elias MC, Sampaio SC, Ciccozzi M, Alcantara LCJ, Slavov SN, Lourenço J, Cella E, and Giovanetti M
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- Humans, Pandemics prevention & control, Genomics, Databases, Factual, Mutation, Genome, Viral, SARS-CoV-2 genetics, COVID-19 epidemiology, COVID-19 prevention & control
- Abstract
The COVID-19 pandemic has brought significant challenges for genomic surveillance strategies in public health systems worldwide. During the past thirty-four months, many countries faced several epidemic waves of SARS-CoV-2 infections, driven mainly by the emergence and spread of novel variants. In that line, genomic surveillance has been a crucial toolkit to study the real-time SARS-CoV-2 evolution, for the assessment and optimization of novel diagnostic assays, and to improve the efficacy of existing vaccines. During the pandemic, the identification of emerging lineages carrying lineage-specific mutations (particularly those in the Receptor Binding domain) showed how these mutations might significantly impact viral transmissibility, protection from reinfection and vaccination. So far, an unprecedented number of SARS-CoV-2 viral genomes has been released in public databases (i.e., GISAID, and NCBI), achieving 14 million genome sequences available as of early-November 2022. In the present review, we summarise the global landscape of SARS-CoV-2 during the first thirty-four months of viral circulation and evolution. It demonstrates the urgency and importance of sustained investment in genomic surveillance strategies to timely identify the emergence of any potential viral pathogen or associated variants, which in turn is key to epidemic and pandemic preparedness., Competing Interests: Declaration of Competing Interest Not declared., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
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- 2023
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22. Retrospective Insights of the COVID-19 Epidemic in the Major Latin American City, São Paulo, Southeastern Brazil.
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Assato PA, Clemente LG, Giovanetti M, Ribeiro G, Lima ARJ, Palmieri M, de Moraes LN, Kashima S, Fukumasu H, Nogueira ML, Alcantara LCJ, Nicolodelli AL, Martins AJ, Petry B, Banho CA, Dos Santos Barros CR, Moncau-Gadbem CT, Moretti DB, De La Roque DGL, Marqueze EC, Mattos EC, Silva FEVD, Da Costa FADS, Cacherik G, De Souza Todao Bernardino J, Lesbon JCC, Sacchetto L, De Lima LPO, Caldeira LAV, Martininghi M, Moraes MM, Poleti MD, Cattony Neto PQ, Cassano RLRC, Brassaloti RA, Slavov SN, Viala VL, Coutinho LL, Grotto RMT, Neto RM, Covas DT, Sampaio SC, Elias MC, and Souza-Neto JA
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- Humans, SARS-CoV-2 genetics, Brazil epidemiology, Latin America, Retrospective Studies, COVID-19 epidemiology
- Abstract
São Paulo is the financial center of Brazil, with a population of over 12 million, that receives travelers from all over the world for business and tourism. It was the first city in Brazil to report a case of COVID-19 that rapidly spread across the city despite the implementation of the restriction measures. Despite many reports, much is still unknown regarding the genomic diversity and transmission dynamics of this virus in the city of São Paulo. Thus, in this study, we provide a retrospective overview of the COVID-19 epidemic in São Paulo City, Southeastern, Brazil, by generating a total of 9995 near-complete genome sequences from all the city's different macro-regions (North, West, Central, East, South, and Southeast). Our analysis revealed that multiple independent introduction events of different variants (mainly Gamma, Delta, and Omicron) occurred throughout time. Additionally, our estimates of viral movement within the different macro-regions further suggested that the East and the Southeast regions were the largest contributors to the Gamma and Delta viral exchanges to other regions. Meanwhile, the North region had a higher contribution to the dispersion of the Omicron variant. Together, our results reinforce the importance of increasing SARS-CoV-2 genomic monitoring within the city and the country to track the real-time evolution of the virus and to detect earlier any eventual emergency of new variants of concern that could undermine the fight against COVID-19 in Brazil and worldwide., Competing Interests: The authors declare no conflict of interest.
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- 2023
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23. Formyl peptide receptors are involved in CTX-induced impairment of lymphocyte functions.
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Zambelli VO, Hösch NG, Farom S, Zychar BC, Spadacci-Morena DD, Carvalho LV, Curi R, Lepsch LB, Scavone C, Sant'Anna OA, Gonçalves LRC, Cury Y, and Sampaio SC
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- Rats, Male, Animals, Rats, Wistar, Receptors, Formyl Peptide metabolism, Endothelial Cells, Lymphocytes, Lipoxygenases metabolism, Lipoxygenases pharmacology, Crotalus metabolism, Crotoxin pharmacology
- Abstract
Crotoxin (CTX) is a neurotoxin that is isolated from the venom of Crotalus durissus terrificus, which displays immunomodulatory, anti-inflammatory, and anti-tumoral effects. Previous research has demonstrated that CTX promotes the adherence of leukocytes to the endothelial cells in blood microcirculation and the high endothelial venules of lymph nodes, which reduces the number of blood cells and lymphocytes. Studies have also shown that these effects are mediated by lipoxygenase-derived mediators. However, the exact lipoxygenase-derived eicosanoid involved in the CTX effect on lymphocytes is yet to be characterized. As CTX stimulates lipoxin-derived mediators from macrophages and lymphocyte effector functions could be modulated by activating formyl peptide receptors, we aimed to investigate whether these receptors were involved in CTX-induced redistribution and functions of lymphocytes in rats. We used male Wistar rats treated with CTX to demonstrate that Boc2 (butoxycarbonyl-Phe-Leu-Phe-Leu-Phe), an antagonist of formyl peptide receptors, prevented CTX-induced decrease in the number of circulating lymphocytes and increased the expression of the lymphocyte adhesion molecule LFA1. CTX reduced the T and B lymphocyte functions, such as lymphocyte proliferation in response to the mitogen Concanavalin A and antibody production in response to BSA immunization, respectively, which was prevented by the administration of Boc2. Importantly, mesenteric lymph node lymphocytes from CTX-treated rats showed an increased release of 15-epi-LXA
4 . These results indicate that formyl peptide receptors mediate CTX-induced redistribution of lymphocytes and that 15-epi-LXA4 is a key mediator of the immunosuppressive effects of CTX., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)- Published
- 2023
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24. A traveling SARS-CoV-2 laboratory as part of a pandemic response among vulnerable Brazilian populations.
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Elias MC, Slavov SN, Lima ARJ, Martins AJ, Barros CRDS, Moretti DB, Araujo EL, Marqueze EC, Ribeiro G, Ribeiro GMF, Bernardino JST, Koser JR, Clemente LG, Crispin LAC, Alcantara LCJ, Coutinho LL, Giovanetti M, Silva QO, Neto RM, Haddad R, Kashima S, Viala VL, Covas DT, and Sampaio SC
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- Humans, COVID-19 Testing, Brazil epidemiology, Pandemics prevention & control, Vulnerable Populations, SARS-CoV-2, COVID-19 diagnosis, COVID-19 epidemiology
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Background: Brazil has been dramatically hit by the SARS-CoV-2 pandemic and is a world leader in COVID-19 morbidity and mortality. Additionally, the largest country of Latin America has been a continuous source of SARS-CoV-2 variants and shows extraordinary variability of the pandemic strains probably related to the country´s outstanding position as a Latin American economical and transportation hub. Not all regions of the country show sufficient infrastructure for SARS-CoV-2 diagnosis and genotyping which can negatively impact the pandemic response., Methods: Due to this reason and to disburden the diagnostic system of the inner São Paulo State, the Butantan Institute established the Mobile Laboratory (in Portuguese: LabMovel) for SARS-CoV-2 testing which started a trip of the most important "hotspots" of the most populous Brazilian region. The LabMovel initiated in two important cities of the State: Aparecida do Norte (an important religious center) and the Baixada Santista region which incorporates the port of Santos, the busiest in Latin America. The LabMovel was fully equipped with an automatized system for SARS-CoV-2 diagnosis and sequencing/genotyping. It also integrated the laboratory systems for patient records and results divulgation including in the Federal Brazilian Healthcare System., Results: Currently,16,678 samples were tested, among them 1,217 from Aparecida and 4,564 from Baixada Santista. We tracked the delta introductio in the tested regions with its high diversification. The established mobile SARS-CoV-2 laboratory had a major impact on the Public Health System of the included cities including timely delivery of the results to the healthcare agents and the Federal Healthcare system, evaluation of the vaccination status of the positive individuals in the background of exponential vaccination process in Brazil and scientific and technological divulgation of the fieldwork to the most vulnerable populations., Conclusions: The SARS-CoV-2 pandemic has demonstrated worldwide the importance of science to fight against this viral agent and the LabMovel shows that it is possible to integrate researchers, clinicians, healthcare workers and patients to take rapid actions that can in fact mitigate this and other epidemiological situations., (© 2022. The Author(s).)
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- 2023
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25. Crotoxin modulates metabolism and secretory activity of peritoneal macrophages from Walker 256 tumor-bearing rats.
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Faiad OJ, Francisco AMSDC, Brigatte P, Curi R, and Sampaio SC
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- Animals, Crotalus metabolism, Cytokines metabolism, Glucose, Glutamine, Hydrogen Peroxide metabolism, Interleukin-6, Macrophages, Peritoneal metabolism, Male, Rats, Rats, Wistar, Tumor Necrosis Factor-alpha, Crotoxin pharmacology
- Abstract
Crotoxin (CTX), the major toxin of Crotalus durissus terrificus snake venom, induces an inhibitory effect on tumor development and modulates the functions of macrophages (MØs), which play a key role as a defense mechanism against tumor growth. In early tumor progression stage, MØs are avidly phagocytic (inflammatory cell), releasing reactive nitrogen intermediates-RNI/ROI and cytokines TNF-α, IL-1β, and IL-6. However, when the tumor has been developed, tumor-associated MØ (angiogenic cell) presents a decrease in the mentioned activities. We reported that CTX stimulates H
2 O2 release, NO production and secretion of cytokines by peritoneal MØs obtained from non-tumor-bearing rats. Considering that the mentioned mediators control tumor growth, it is mandatory to investigate whether CTX stimulates the production of these mediators by MØs obtained from tumor-bearing animals. The aim of this work was then to evaluate the CTX effect on metabolism and functions of peritoneal MØs obtained from Walker 256 tumor-bearing rats. For this purpose, male Wistar rats were subcutaneously inoculated in the right flank with 1 mL sterile suspension of 2 × 107 Walker 256 tumor cells. CTX (18 μg per animal) was subcutaneously administered in two protocols: a) on the 1st day of tumor cell injection and b) on the 4th day of tumor cell inoculation. In both protocols, MØs were obtaining on the 14th day of tumor cell inoculation to evaluate the release of H2 O2, NO, and pro-inflammatory cytokines (IL-1β, TNFα, and IL-6); maximal activity of hexokinase, glucose-6-phosphate dehydrogenase, citrate synthase, and14 CO2 production from [U-14 C]-glucose and [U-14 C]-glutamine. The treatment with CTX stimulated the release of NO, H2 O2 , and cytokines, and glucose and glutamine metabolism. Metabolic and functional changes induced by CTX were accompanied by a decrease of tumor growth as indicated by tumor fresh weight and diameter. These results indicate CTX not only as a scientific tool to investigate changes in metabolism and functions of peritoneal MØs but also for a better understanding of the mechanisms involved in tumor growth., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)- Published
- 2022
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26. Dynamics of SARS-CoV-2 Variants of Concern in Vaccination Model City in the State of Sao Paulo, Brazil.
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Slavov SN, de La-Roque DGL, da Costa PNM, Rodrigues ES, Santos EV, Borges JS, Evaristo M, de Matos Maçonetto J, Marques AA, Milhomens J, Rós FA, Fonseca V, Lima ARJ, Ribeiro G, Lima LPO, Garibaldi PMM, Ferreira NN, Moraes GR, Marqueze EC, Barros CRDS, Martins AJ, Coutinho LL, Calado RT, Borges M, Elias MC, Sampaio SC, Giovanetti M, Alcantara LCJ, Covas DT, and Kashima S
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- Humans, Brazil epidemiology, Phylogeny, Vaccination, SARS-CoV-2 genetics, COVID-19 epidemiology, COVID-19 prevention & control
- Abstract
From a country with one of the highest SARS-CoV-2 morbidity and mortality rates, Brazil has implemented one of the most successful vaccination programs. Brazil's first model city vaccination program was performed by the CoronaVac vaccine (Sinovac Biotech) in the town of Serrana, São Paulo State. To evaluate the vaccination effect on the SARS-CoV-2 molecular dynamics and clinical outcomes, we performed SARS-CoV-2 molecular surveillance on 4375 complete genomes obtained between June 2020 and April 2022 in this location. This study included the period between the initial SARS-CoV-2 introduction and during the vaccination process. We observed that the SARS-CoV-2 substitution dynamics in Serrana followed the viral molecular epidemiology in Brazil, including the initial identification of the ancestral lineages (B.1.1.28 and B.1.1.33) and epidemic waves of variants of concern (VOC) including the Gamma, Delta, and, more recently, Omicron. Most probably, as a result of the immunization campaign, the mortality during the Gamma and Delta VOC was significantly reduced compared to the rest of Brazil, which was also related to lower morbidity. Our phylogenetic analysis revealed the evolutionary history of the SARS-CoV-2 in this location and showed that multiple introduction events have occurred over time. The evaluation of the COVID-19 clinical outcome revealed that most cases were mild (88.9%, 98.1%, 99.1% to Gamma, Delta, and Omicron, respectively) regardless of the infecting VOC. In conclusion, we observed that vaccination was responsible for reducing the death toll rate and related COVID-19 morbidity, especially during the gamma and Delta VOC; however, it does not prevent the rapid substitution rate and morbidity of the Omicron VOC.
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- 2022
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27. Correction: Lesbon et al. Nucleocapsid (N) Gene Mutations of SARS-CoV-2 Can Affect Real-Time RT-PCR Diagnostic and Impact False-Negative Results. Viruses 2021, 13 , 2474.
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Lesbon JCC, Poleti MD, de Mattos Oliveira EC, Patané JSL, Clemente LG, Viala VL, Ribeiro G, Giovanetti M, de Alcantara LCJ, Teixeira O, Nonato MC, de Lima LPO, Martins AJ, Dos Santos Barros CR, Marqueze EC, de Souza Todão Bernardino J, Moretti DB, Brassaloti RA, de Lello Rocha Campos Cassano R, Mariani PDSC, Slavov SN, Dos Santos RB, Rodrigues ES, Santos EV, Borges JS, de La Roque DGL, Kitajima JP, Santos B, Assato PA, da Silva da Costa FA, Banho CA, Sacchetto L, Moraes MM, Palmieri M, da Silva FEV, Grotto RMT, Souza-Neto JA, Nogueira ML, Coutinho LL, Calado RT, Neto RM, Covas DT, Kashima S, Elias MC, Sampaio SC, and Fukumasu H
- Abstract
The authors hereby request the inclusion of two authors (Olivia Teixeira and Maria Cristina Nonato) in the recently published article in Viruses entitled "Nucleocapsid (N) gene mutations of SARS-CoV-2 can affect real-time RT-PCR diagnostic and impact false-negative results" [...].
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- 2022
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28. Genomic epidemiology of the SARS-CoV-2 epidemic in Brazil.
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Giovanetti M, Slavov SN, Fonseca V, Wilkinson E, Tegally H, Patané JSL, Viala VL, San EJ, Rodrigues ES, Santos EV, Aburjaile F, Xavier J, Fritsch H, Adelino TER, Pereira F, Leal A, Iani FCM, de Carvalho Pereira G, Vazquez C, Sanabria GME, Oliveira EC, Demarchi L, Croda J, Dos Santos Bezerra R, Paola Oliveira de Lima L, Martins AJ, Renata Dos Santos Barros C, Marqueze EC, de Souza Todao Bernardino J, Moretti DB, Brassaloti RA, de Lello Rocha Campos Cassano R, Mariani PDSC, Kitajima JP, Santos B, Proto-Siqueira R, Cantarelli VV, Tosta S, Nardy VB, Reboredo de Oliveira da Silva L, Gómez MKA, Lima JG, Ribeiro AA, Guimarães NR, Watanabe LT, Barbosa Da Silva L, da Silva Ferreira R, da Penha MPF, Ortega MJ, de la Fuente AG, Villalba S, Torales J, Gamarra ML, Aquino C, Figueredo GPM, Fava WS, Motta-Castro ARC, Venturini J, do Vale Leone de Oliveira SM, Gonçalves CCM, do Carmo Debur Rossa M, Becker GN, Giacomini MP, Marques NQ, Riediger IN, Raboni S, Mattoso G, Cataneo AD, Zanluca C, Duarte Dos Santos CN, Assato PA, Allan da Silva da Costa F, Poleti MD, Lesbon JCC, Mattos EC, Banho CA, Sacchetto L, Moraes MM, Grotto RMT, Souza-Neto JA, Nogueira ML, Fukumasu H, Coutinho LL, Calado RT, Neto RM, Bispo de Filippis AM, Venancio da Cunha R, Freitas C, Peterka CRL, de Fátima Rangel Fernandes C, Navegantes W, do Carmo Said RF, Campelo de A E Melo CF, Almiron M, Lourenço J, de Oliveira T, Holmes EC, Haddad R, Sampaio SC, Elias MC, Kashima S, Junior de Alcantara LC, and Covas DT
- Subjects
- Brazil, Genomics, Humans, COVID-19, SARS-CoV-2
- Abstract
The high numbers of COVID-19 cases and deaths in Brazil have made Latin America an epicentre of the pandemic. SARS-CoV-2 established sustained transmission in Brazil early in the pandemic, but important gaps remain in our understanding of virus transmission dynamics at a national scale. We use 17,135 near-complete genomes sampled from 27 Brazilian states and bordering country Paraguay. From March to November 2020, we detected co-circulation of multiple viral lineages that were linked to multiple importations (predominantly from Europe). After November 2020, we detected large, local transmission clusters within the country. In the absence of effective restriction measures, the epidemic progressed, and in January 2021 there was emergence and onward spread, both within and abroad, of variants of concern and variants under monitoring, including Gamma (P.1) and Zeta (P.2). We also characterized a genomic overview of the epidemic in Paraguay and detected evidence of importation of SARS-CoV-2 ancestor lineages and variants of concern from Brazil. Our findings show that genomic surveillance in Brazil enabled assessment of the real-time spread of emerging SARS-CoV-2 variants., (© 2022. The Author(s).)
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- 2022
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29. Crotoxin modulates inflammation and macrophages' functions in a murine sepsis model.
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Bretones ML, Sampaio SC, Barbeiro DF, Ariga SKK, Soriano FG, and Lima TM
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- Animals, Crotalus, Escherichia coli, Hydrogen Peroxide pharmacology, Inflammation, Interleukin-10, Interleukin-6, Macrophages, Male, Mice, Mice, Inbred BALB C, Crotoxin pharmacology, Sepsis drug therapy
- Abstract
Sepsis is a syndrome of physiological and biochemical abnormalities induced by an infection that represents a major public health concern. It involves the early activation of inflammatory responses. Crotoxin (CTX), the major toxin of the South American rattlesnake Crotalus durissus terrificus venom, presents longstanding anti-inflammatory properties. Since immune system modulation may be a strategic target in sepsis management, and macrophages' functional and secretory activities are related to the disease's progression, we evaluated the effects of CTX on macrophages from septic animals. Balb/c male mice submitted to cecal ligation and puncture (CLP) were treated with CTX (0.9 μg/animal, subcutaneously) 1 h after the procedure and euthanized after 6 h. We used plasma samples to quantify circulating cytokines and eicosanoids. Bone marrow differentiated macrophages (BMDM) were used to evaluate the CTX effect on macrophages' functions. Our data show that CTX administration increased the survival rate of the animals from 40% to 80%. Septic mice presented lower plasma concentrations of IL-6 and TNF-α after CTX treatment, and higher concentrations of LXA
4 , PGE2, and IL-1β. No effect was observed in IL-10, IFN-γ, and RD1 concentrations. BMDM from septic mice treated with CTX presented decreased capacity of E. coli phagocytosis, but sustained NO and H2 O2 production. We also observed higher IL-6 concentration in the culture medium of BMDM from septic mice, and CTX induced a significant reduction. CTX treatment increased IL-10 production by macrophages as well. Our data show that the protective effect of CTX in sepsis mortality involves modulation of macrophage functions and inflammatory mediators' production., (Copyright © 2022 Elsevier Ltd. All rights reserved.)- Published
- 2022
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30. Viral Metagenomics for the Identification of Emerging Infections in Clinical Samples with Inconclusive Dengue, Zika, and Chikungunya Viral Amplification.
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Souza JVC, Santos HO, Leite AB, Giovanetti M, Bezerra RDS, Carvalho E, Bernardino JST, Viala VL, Haddad R, Ciccozzi M, Alcantara LCJ, Sampaio SC, Covas DT, Kashima S, Elias MC, and Slavov SN
- Subjects
- Arboviruses genetics, Communicable Diseases, Emerging, Humans, Metagenomics, Phylogeny, Real-Time Polymerase Chain Reaction, Chikungunya Fever diagnosis, Chikungunya Fever epidemiology, Chikungunya virus genetics, Dengue diagnosis, Dengue epidemiology, Dengue Virus genetics, Zika Virus genetics, Zika Virus Infection
- Abstract
Viral metagenomics is increasingly being used for the identification of emerging and re-emerging viral pathogens in clinical samples with unknown etiology. The objective of this study was to shield light on the metavirome composition in clinical samples obtained from patients with clinical history compatible with an arboviral infection, but that presented inconclusive results when tested using RT-qPCR. The inconclusive amplification results might be an indication of the presence of an emerging arboviral agent that is inefficiently amplified by conventional PCR techniques. A total of eight serum samples with inconclusive amplification results for the routinely tested arboviruses-dengue (DENV), Zika (ZIKV), and Chikungunya (CHIKV) obtained during DENV and CHIKV outbreaks registered in the state of Alagoas, Northeast Brazil between July and August 2021-were submitted to metagenomic next-generation sequencing assay using NextSeq 2000 and bioinformatic pipeline for viral discovery. The performed bioinformatic analysis revealed the presence of two arboviruses: DENV type 2 (DENV-2) and CHIKV with a high genome coverage. Further, the metavirome of those samples revealed the presence of multiple commensal viruses apparently without clinical significance. The phylogenetic analysis demonstrated that the DENV-2 genome belonged to the Asian/American genotype and clustered with other Brazilian strains. The identified CHIKV genome was taxonomically assigned as ECSA genotype, which is circulating in Brazil. Together, our results reinforce the utility of metagenomics as a valuable tool for viral identification in samples with inconclusive arboviral amplification. Viral metagenomics is one of the most potent methods for the identification of emerging arboviruses.
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- 2022
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31. The Divergent Pattern of SARS-CoV-2 Variant Predominance and Transmission Dynamics in the Brazilian Island of Ilhabela.
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Viala VL, Slavov SN, de Lima LPO, Lima ARJ, Ribeiro G, Martins AJ, Petry B, Banho CA, Barros CRDS, Moncau CT, Moretti DB, de La-Roque DGL, Marqueze EC, Mattos EC, Costa FADSD, Fukumasu H, Bernardino JST, Souza-Neto JA, Lesbon JCC, Kayanoki LP, Bernardo LL, Sacchetto L, Clemente LG, Alcantara LCJ, Coutinho LL, Marques BC, Giovanetti M, Nogueira ML, Poleti MD, Assato PA, Cattony Neto PQ, Cassano RLRC, Neto RM, Grotto RMT, Brassaloti RA, Kashima S, Covas DT, Elias MC, and Sampaio SC
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- Brazil epidemiology, Humans, Phylogeny, COVID-19 epidemiology, SARS-CoV-2 genetics
- Abstract
Our effort in SARS-CoV-2 genomic surveillance in Brazil has detected the Alpha Variant of Concern with a predominance higher than 75% in the population of Ilhabela island (São Paulo State) at a time when the Gamma VOC was already predominating the mainland raised concerns for closer surveillance on this island. Therefore, we intensified the surveillance for 24 weeks by generating data from 34% of local positive cases. Our data show that the patterns of VOC predominance dynamics and infection rates were in general distinct from the mainland. We report here the first known case of Alpha predominance in a Brazilian population, a delay greater than 3 months for the Gamma to dominate the previous variants compared to the mainland, and a faster dispersion rate of Gamma and Delta VOCs compared to the mainland. Phylogenetic analysis revealed the SARS-CoV-2 transmission dynamics in Ilhabela were characterized by multiple independent introduction events of Gamma and Delta, with a few events of Alpha introduction, two of them followed by community transmission. This study evidenced the peculiar behavior of SARS-CoV-2 variants in an isolated population and brought to light the importance of specific programs for SARS-CoV-2 genomic surveillance in isolated populations.
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- 2022
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32. SARS-CoV-2 epidemic in Brazil: how the displacement of variants has driven distinct epidemic waves.
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Alcantara LCJ, Nogueira E, Shuab G, Tosta S, Fristch H, Pimentel V, Souza-Neto JA, Coutinho LL, Fukumasu H, Sampaio SC, Elias MC, Kashima S, Slavov SN, Ciccozzi M, Cella E, Lourenco J, Fonseca V, and Giovanetti M
- Subjects
- Brazil epidemiology, Humans, Mutation, Spike Glycoprotein, Coronavirus genetics, COVID-19 epidemiology, SARS-CoV-2 genetics
- Abstract
Brazil ranks as third in terms of total number of reported SARS-CoV-2 cases globally. The COVID-19 epidemic in Brazil was characterised by the co-circulation of multiple variants as a consequence of multiple independent introduction events occurring through time. Here, we describe the SARS-CoV-2 variants that are currently circulating and co-circulating in the country, with the aim to highlight which variants have driven the different epidemic waves. For this purpose, we retrieved metadata information of Coronavirus sequences collected in Brazil and available at the GISAID database. SARS-CoV-2 lineages have been identified along with eleven variants, labelled as VOCs (Alpha, Gamma, Beta, Delta and Omicron) VOIs (Lambda and Mu) VUMs (B.1.1.318) and FMVs (Zeta, Eta and B.1.1.519). Here we show that, in the Brazilian context, after 24 months of sustained transmission and evolution of SARS-CoV-2, local variants (among them the B.1.1.28 and B.1.1.33) were displaced by recently introduced VOCs firstly with the Gamma, followed by Delta and more recently Omicron. The rapid spread of some of those VOCs (such as Gamma and Omicron) was also mirror by a large increase in the number of cases and deaths in the country. This in turn reinforces that, due to the emergence of variants that appear to induce a substantial evasion against neutralizing antibody response, it is important to strengthen genomic effort within the country and how vaccination still remains a critical process to protect the vulnerable population, still at risk of infection and death., (Copyright © 2022. Published by Elsevier B.V.)
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- 2022
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33. SARS-COV-2 genomic monitoring in the state of São Paulo unveils two emerging AY.43 sublineages.
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Lima ARJ, Ribeiro G, Viala VL, de Lima LPO, Martins AJ, Barros CRDS, Marqueze EC, Bernardino JST, Moretti DB, Rodrigues ES, Santos EV, Brassaloti RA, Cassano RLRC, Mariani PDSC, Clemente LG, Assato PA, Costa FADSD, Poleti MD, Lesbon JCC, Mattos EC, Banho CA, Sacchetto L, Moraes MM, Palmieri M, Martininghi M, Caldeira LAV, Silva FEVD, Grotto RMT, Souza-Neto JA, Giovanetti M, Junior Alcantara LC, Nogueira ML, Fukumasu H, Coutinho LL, Kashima S, Neto RM, Covas DT, Slavov SN, Sampaio SC, and Elias MC
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- Brazil epidemiology, COVID-19 Vaccines, Genomics, Humans, COVID-19 epidemiology, SARS-CoV-2 genetics
- Abstract
Delta VOC is highly diverse with more than 120 sublineages already described as of November 30, 2021. In this study, through active monitoring of circulating severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) variants in the state of São Paulo, southeast Brazil, we identified two emerging sublineages from the ancestral AY.43 strain which were classified as AY.43.1 and AY.43.2. These sublineages were defined by the following characteristic nonsynonymous mutations ORF1ab:A4133V and ORF3a:T14I for the AY.43.1 and ORF1ab:G1155C for the AY.43.2 and our analysis reveals that they might have a likely-Brazilian origin. Much is still unknown regarding their dissemination in the state of São Paulo and Brazil as well as their potential impact on the ongoing vaccination process. However, the results obtained in this study reinforce the importance of genomic surveillance activity for timely identification of emerging SARS-CoV-2 variants which can impact the ongoing SARS-CoV-2 vaccination and public health policies., (© 2022 Wiley Periodicals LLC.)
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- 2022
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34. Genomic epidemiology reveals the impact of national and international restrictions measures on the SARS-CoV-2 epidemic in Brazil.
- Author
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Giovanetti M, Slavov SN, Fonseca V, Wilkinson E, Tegally H, Patané JSL, Viala VL, San JE, Rodrigues ES, Santos EV, Aburjaile F, Xavier J, Fritsch H, Adelino TER, Pereira F, Leal A, de Melo Iani FC, de Carvalho Pereira G, Vazquez C, Mercedes Estigarribia Sanabria G, de Oliveira EC, Demarchi L, Croda J, Dos Santos Bezerra R, de Lima LPO, Martins AJ, Dos Santos Barros CR, Marqueze EC, de Souza Todao Bernardino J, Moretti DB, Brassaloti RA, de Lello Rocha Campos Cassano R, Mariani PDSC, Kitajima JP, Santos B, Proto-Siqueira R, Cantarelli VV, Tosta S, Nardy VB, de Oliveira da Silva LR, Kelly Astete Gómez M, Lima JG, Ribeiro AA, Guimarães NR, Watanabe LT, Da Silva LB, da Silva Ferreira R, da Penha MPF, Ortega MJ, de la Fuente AG, Villalba S, Torales J, Gamarra ML, Aquino C, Martínez Figueredo GP, Fava WS, Motta-Castro ARC, Venturini J, de Oliveira SMDVL, Gonçalves CCM, do Carmo Debur Rossa M, Becker GN, Presibella MM, Marques NQ, Riediger IN, Raboni S, Coelho GM, Cataneo AHD, Zanluca C, Dos Santos CND, Assato PA, da Costa FADS, Poleti MD, Lesbon JCC, Mattos EC, Banho CA, Sacchetto L, Moraes MM, Grotto RMT, Souza-Neto JA, Nogueira ML, Fukumasu H, Coutinho LL, Calado RT, Neto RM, de Filippis AMB, da Cunha RV, Freitas C, Peterka CRL, de Fátima Rangel Fernandes C, de Araújo WN, do Carmo Said RF, Almiron M, de Albuquerque E Melo CFC, Lourenço J, de Oliveira T, Holmes EC, Haddad R, Sampaio SC, Elias MC, Kashima S, de Alcantara LCJ, and Covas DT
- Abstract
Brazil has experienced some of the highest numbers of COVID-19 cases and deaths globally and from May 2021 made Latin America a pandemic epicenter. Although SARS-CoV-2 established sustained transmission in Brazil early in the pandemic, important gaps remain in our understanding of virus transmission dynamics at the national scale. Here, we describe the genomic epidemiology of SARS-CoV-2 using near-full genomes sampled from 27 Brazilian states and a bordering country - Paraguay. We show that the early stage of the pandemic in Brazil was characterised by the co-circulation of multiple viral lineages, linked to multiple importations predominantly from Europe, and subsequently characterized by large local transmission clusters. As the epidemic progressed under an absence of effective restriction measures, there was a local emergence and onward international spread of Variants of Concern (VOC) and Variants Under Monitoring (VUM), including Gamma (P.1) and Zeta (P.2). In addition, we provide a preliminary genomic overview of the epidemic in Paraguay, showing evidence of importation from Brazil. These data reinforce the usefulness and need for the implementation of widespread genomic surveillance in South America as a toolkit for pandemic monitoring that provides a means to follow the real-time spread of emerging SARS-CoV-2 variants with possible implications for public health and immunization strategies.
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- 2022
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35. Comparison between the rotary (Hyflex EDM®) and manual (k-file) technique for instrumentation of primary molars: a 12-month randomized clinical follow-up study.
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Amorim AC, Caldeira AV, Sampaio SC, Lourenço Neto N, Oliveira TM, Nogueira DA, Moretti ABDS, and Sakai VT
- Subjects
- Child, Follow-Up Studies, Humans, Tooth, Deciduous, Molar diagnostic imaging, Molar surgery, Root Canal Preparation
- Abstract
Objectives: Rotational instrumentation is an alternative for the clinical practice of pediatric dentists. However, there are few records in the literature on the clinical and radiographic aspects of treated teeth over time. Compare instrumentation time and filling quality between manual (k-file) and rotary (Hyflex EDM®) files, and clinically and radiographically follow-up the treated teeth for 12 months. Moreover, the characteristics of glass ionomer restorations and their interference in the treatment prognosis over time were evaluated., Methodology: In total, 40 children with pulp involvement in primary molars received treatment with Hyflex EDM® or manual rotary files, performed by an operator. Clinical and radiographic aspects were observed at different times to determine the effectiveness of each technique., Results: The rotary system reduced instrumentation time when compared to the use of manual files (p≤0.05), but there was no difference in filling quality between the groups (p≥0.05). Moreover, both types of instrumentation were effective for 12 months (p≥0.05), and restoration retention influenced the emergence of periapical lesions (p≤0.05)., Conclusion: Although rotary files reduce clinical time, the clinical and radiographic aspects of both techniques were similar over 12 months. Moreover, restoration retention has been shown to be related to treatment prognosis.
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- 2022
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36. Introduction of SARS-CoV-2 C.37 (WHO VOI lambda) in the Sao Paulo State, Southeast Brazil.
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Kashima S, Slavov SN, Giovanetti M, Rodrigues ES, Patané JSL, Viala VL, Santos EV, Evaristo M, de Lima LPO, Martins AJ, Dos Santos Barros CR, Marqueze EC, Garibaldi PMM, Ferreira NN, Moraes GR, Brassaloti RA, Cassano RLRC, Mariani PDSC, Kitajima JP, Schlesinger D, Bezerra RS, Assato PA, da Costa FAS, Poleti MD, Lesbon JCC, Mattos EC, Banho CA, Sacchetto L, Grotto RMT, Souza-Neto JA, Fonseca V, de Alcantara LCJ, Nogueira ML, Fukumasu H, Coutinho LL, Borges M, Calado RT, Elias MC, Sampaio SC, and Covas DT
- Subjects
- Brazil epidemiology, Humans, World Health Organization, COVID-19 epidemiology, SARS-CoV-2 genetics
- Abstract
The Lambda variants of interest (VOI) (C37/GR/452Q.V1/21G) was initially reported in Lima, Peru but has gained rapid dissemination through other Latin American countries. Nevertheless, the dissemination and molecular epidemiology of the Lambda VOI in Brazil is unknown apart from a single case report. In this respect, we characterized the circulation of the SARS-CoV-2 Lambda VOI (C37/GR/452Q.V1/21G) in Sao Paulo State, Brazil. From March to June 2021, we identified seven Lambda isolates in a set of approximately 8000 newly sequenced genomes of the Network for Pandemic Alert of Emerging SARS-CoV-2 variants from Sao Paulo State. Interestingly, in three of the positive patients, the Lambda VOI infection was probably related to a contact transmission. These individuals were fully vaccinated to COVID-19 and presented mild symptoms. The remaining positive for Lambda VOI individuals showed different levels of COVID-19 symptoms and one of them needed hospitalization (score 5, WHO). In our study, we present a low level of Lambda VOI circulation in the Sao Paulo State. This reinforces the essential role of molecular surveillance for the effective SARS-CoV-2 pandemic response, especially in regard to circulating variants., (© 2021 Wiley Periodicals LLC.)
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- 2022
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37. Human Sensory Neuron-like Cells and Glycated Collagen Matrix as a Model for the Screening of Analgesic Compounds.
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Bufalo MC, Almeida MES, Jensen JR, DeOcesano-Pereira C, Lichtenstein F, Picolo G, Chudzinski-Tavassi AM, Sampaio SC, Cury Y, and Zambelli VO
- Subjects
- Animals, Antigens, Neoplasm metabolism, Biomarkers metabolism, Cell Differentiation drug effects, Cell Line, Tumor, Cell Survival drug effects, Extracellular Matrix metabolism, Galectin 3 metabolism, Gene Expression Regulation drug effects, Glycosylation drug effects, Humans, Mitogen-Activated Protein Kinases metabolism, NAV1.7 Voltage-Gated Sodium Channel genetics, NAV1.7 Voltage-Gated Sodium Channel metabolism, Neurites drug effects, Neurites metabolism, Neurons cytology, Neurons drug effects, Proto-Oncogene Proteins c-fos metabolism, Rats, Receptors, Neurokinin-1 genetics, Receptors, Neurokinin-1 metabolism, Sensory Receptor Cells drug effects, Sensory Receptor Cells metabolism, Substance P metabolism, beta-Endorphin metabolism, Analgesics analysis, Analgesics pharmacology, Collagen pharmacology, Drug Evaluation, Preclinical, Models, Biological, Sensory Receptor Cells cytology
- Abstract
Increased collagen-derived advanced glycation end-products (AGEs) are consistently related to painful diseases, including osteoarthritis, diabetic neuropathy, and neurodegenerative disorders. We have recently developed a model combining a two-dimensional glycated extracellular matrix (ECM-GC) and primary dorsal root ganglion (DRG) that mimicked a pro-nociceptive microenvironment. However, culturing primary cells is still a challenge for large-scale screening studies. Here, we characterized a new model using ECM-GC as a stimulus for human sensory-like neurons differentiated from SH-SY5Y cell lines to screen for analgesic compounds. First, we confirmed that the differentiation process induces the expression of neuron markers (MAP2, RBFOX3 (NeuN), and TUBB3 (β-III tubulin), as well as sensory neuron markers critical for pain sensation (TRPV1, SCN9A (Nav1.7), SCN10A (Nav1.8), and SCN11A (Nav1.9). Next, we showed that ECM-GC increased c-Fos expression in human sensory-like neurons, which is suggestive of neuronal activation. In addition, ECM-GC upregulated the expression of critical genes involved in pain, including SCN9A and TACR1 . Of interest, ECM-GC induced substance P release, a neuropeptide widely involved in neuroinflammation and pain. Finally, morphine, the prototype opiate, decreased ECM-GC-induced substance P release. Together, our results suggest that we established a functional model that can be useful as a platform for screening candidates for the management of painful conditions.
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- 2022
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38. A Sequencing Network that São Paulo State population deserves: a successful case of supporting Public Health.
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Elias MC, Ranieri A, Ribeiro G, Covas DT, and Sampaio SC
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- 2022
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39. Nucleocapsid (N) Gene Mutations of SARS-CoV-2 Can Affect Real-Time RT-PCR Diagnostic and Impact False-Negative Results.
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Lesbon JCC, Poleti MD, de Mattos Oliveira EC, Patané JSL, Clemente LG, Viala VL, Ribeiro G, Giovanetti M, de Alcantara LCJ, Teixeira O, Nonato MC, de Lima LPO, Martins AJ, Dos Santos Barros CR, Marqueze EC, de Souza Todão Bernardino J, Moretti DB, Brassaloti RA, de Lello Rocha Campos Cassano R, Mariani PDSC, Slavov SN, Dos Santos RB, Rodrigues ES, Santos EV, Borges JS, de La Roque DGL, Kitajima JP, Santos B, Assato PA, da Silva da Costa FA, Banho CA, Sacchetto L, Moraes MM, Palmieri M, da Silva FEV, Grotto RMT, Souza-Neto JA, Nogueira ML, Coutinho LL, Calado RT, Neto RM, Covas DT, Kashima S, Elias MC, Sampaio SC, and Fukumasu H
- Subjects
- Brazil epidemiology, COVID-19 epidemiology, Coronavirus RNA-Dependent RNA Polymerase genetics, DNA Primers, False Negative Reactions, Genome, Viral genetics, Humans, Mutation, Phosphoproteins genetics, RNA, Viral genetics, SARS-CoV-2 genetics, COVID-19 diagnosis, COVID-19 Nucleic Acid Testing, Coronavirus Nucleocapsid Proteins genetics, SARS-CoV-2 isolation & purification
- Abstract
The current COVID-19 pandemic demands massive testing by Real-time RT-PCR (Reverse Transcription Polymerase Chain Reaction), which is considered the gold standard diagnostic test for the detection of the SARS-CoV-2 virus. However, the virus continues to evolve with mutations that lead to phenotypic alterations as higher transmissibility, pathogenicity or vaccine evasion. Another big issue are mutations in the annealing sites of primers and probes of RT-PCR diagnostic kits leading to false-negative results. Therefore, here we identify mutations in the N (Nucleocapsid) gene that affects the use of the GeneFinder COVID-19 Plus RealAmp Kit. We sequenced SARS-CoV-2 genomes from 17 positive samples with no N gene detection but with RDRP (RNA-dependent RNA polymerase) and E (Envelope) genes detection, and observed a set of three different mutations affecting the N detection: a deletion of 18 nucleotides (Del28877-28894), a substitution of GGG to AAC (28881-28883) and a frameshift mutation caused by deletion (Del28877-28878). The last one cause a deletion of six AAs (amino acids) located in the central intrinsic disorder region at protein level. We also found this mutation in 99 of the 14,346 sequenced samples by the Sao Paulo state Network for Pandemic Alert of Emerging SARS-CoV-2 variants, demonstrating the circulation of the mutation in Sao Paulo, Brazil. Continuous monitoring and characterization of mutations affecting the annealing sites of primers and probes by genomic surveillance programs are necessary to maintain the effectiveness of the diagnosis of COVID-19.
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- 2021
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40. Correlating biological activity to thermo-structural analysis of the interaction of CTX with synthetic models of macrophage membranes.
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de Araújo Pimenta L, Duarte EL, Muniz GSV, Pasqualoto KFM, de Mattos Fontes MR, Lamy MT, and Sampaio SC
- Subjects
- Algorithms, Animals, Crotalus, Fluorescent Antibody Technique, Humans, Kinetics, Models, Theoretical, Molecular Structure, Protein Binding, Spectrum Analysis, Structure-Activity Relationship, THP-1 Cells, Cell Membrane chemistry, Cell Membrane metabolism, Crotoxin chemistry, Crotoxin metabolism, Macrophages metabolism, Thermodynamics
- Abstract
The important pharmacological actions of Crotoxin (CTX) on macrophages, the main toxin in the venom of Crotalus durissus terrificus, and its important participation in the control of different pathophysiological processes, have been demonstrated. The biological activities performed by macrophages are related to signaling mediated by receptors expressed on the membrane surface of these cells or opening and closing of ion channels, generation of membrane curvature and pore formation. In the present work, the interaction of the CTX complex with the cell membrane of macrophages is studied, both using biological cells and synthetic lipid membranes to monitor structural alterations induced by the protein. Here we show that CTX can penetrate THP-1 cells and induce pores only in anionic lipid model membranes, suggesting that a possible access pathway for CTX to the cell is via lipids with anionic polar heads. Considering that the selectivity of the lipid composition varies in different tissues and organs of the human body, the thermostructural studies presented here are extremely important to open new investigations on the biological activities of CTX in different biological systems., (© 2021. The Author(s).)
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- 2021
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41. Genomic monitoring unveil the early detection of the SARS-CoV-2 B.1.351 (beta) variant (20H/501Y.V2) in Brazil.
- Author
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Slavov SN, Patané JSL, Bezerra RDS, Giovanetti M, Fonseca V, Martins AJ, Viala VL, Rodrigues ES, Santos EV, Barros CRS, Marqueze EC, Santos B, Aburjaile F, Neto RM, Moretti DB, Haddad R, Calado RT, Kitajima JP, Freitas E, Schlesinger D, Junior de Alcantara LC, Elias MC, Sampaio SC, Kashima S, and Covas DT
- Subjects
- Antibodies, Neutralizing immunology, Antibodies, Viral immunology, Brazil, COVID-19 immunology, COVID-19 virology, Genomics methods, Humans, Mutation genetics, Mutation immunology, SARS-CoV-2 immunology, Spike Glycoprotein, Coronavirus genetics, Spike Glycoprotein, Coronavirus immunology, SARS-CoV-2 genetics
- Abstract
Sao Paulo State, currently experiences a second COVID-19 wave overwhelming the healthcare system. Due to the paucity of SARS-CoV-2 complete genome sequencing, we established a Network for Pandemic Alert of Emerging SARS-CoV-2 Variants to rapidly understand and monitor the spread of SARS-CoV-2 variants into the state. Through analysis of 210 SARS-CoV-2 complete genomes obtained from the largest regional health departments we identified cocirculation of multiple SARS-CoV-2 lineages such as B.1.1 (0.5%), B.1.1.28 (23.2%), B.1.1.7 (alpha variant, 6.2%), B.1.566 (1.4%), B.1.544 (0.5%), C.37 (0.5%) P.1 (gamma variant, 66.2%), and P.2 (zeta variant, 1.0%). Our analysis allowed also the detection, for the first time in Brazil, the South African B.1.351 (beta) variant of concern, B.1.351 (501Y.V2) (0.5%), characterized by the following mutations: ORF1ab: T265I, R724K, S1612L, K1655N, K3353R, SGF 3675_F3677del, P4715L, E5585D; spike: D80A, D215G, L242_L244del, A262D, K417N, E484K, N501Y, D614G, A701V, C1247F; ORF3a: Q57H, S171L, E: P71L; ORF7b: Y10F, N: T205I; ORF14: L52F. The most recent common ancestor of the identified strain was inferred to be mid-October to late December 2020. Our analysis demonstrated the P.1 lineage predominance and allowed the early detection of the South African strain for the first time in Brazil. We highlight the importance of SARS-CoV-2 active monitoring to ensure the rapid detection of potential variants for pandemic control and vaccination strategies. Highlights Identification of B.1.351 (beta) variant of concern in the Sao Paulo State. Dissemination of SARS-CoV-2 variants of concern and interest in the Sao Paulo State. Mutational Profile of the circulating variants of concern and interest., (© 2021 Wiley Periodicals LLC.)
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- 2021
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42. Crotoxin Modulates Events Involved in Epithelial-Mesenchymal Transition in 3D Spheroid Model.
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Kato EE and Sampaio SC
- Subjects
- A549 Cells, Antineoplastic Agents isolation & purification, Antineoplastic Agents pharmacology, Cell Line, Collagen Type I metabolism, Crotalid Venoms chemistry, Crotoxin isolation & purification, Fibroblasts drug effects, Fibroblasts metabolism, Humans, Spheroids, Cellular metabolism, Crotoxin pharmacology, Epithelial-Mesenchymal Transition drug effects, Spheroids, Cellular drug effects, Tumor Microenvironment drug effects
- Abstract
Epithelial-mesenchymal transition (EMT) occurs in the early stages of embryonic development and plays a significant role in the migration and the differentiation of cells into various types of tissues of an organism. However, tumor cells, with altered form and function, use the EMT process to migrate and invade other tissues in the body. Several experimental (in vivo and in vitro) and clinical trial studies have shown the antitumor activity of crotoxin (CTX), a heterodimeric phospholipase A2 present in the Crotalus durissus terrificus venom. In this study, we show that CTX modulates the microenvironment of tumor cells. We have also evaluated the effect of CTX on the EMT process in the spheroid model. The invasion of type I collagen gels by heterospheroids (mix of MRC-5 and A549 cells constitutively prepared with 12.5 nM CTX), expression of EMT markers, and secretion of MMPs were analyzed. Western blotting analysis shows that CTX inhibits the expression of the mesenchymal markers, N-cadherin, α-SMA, and αv. This study provides evidence of CTX as a key modulator of the EMT process, and its antitumor action can be explored further for novel drug designing against metastatic cancer.
- Published
- 2021
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43. Crotoxin Inhibits Endothelial Cell Functions in Two- and Three-dimensional Tumor Microenvironment.
- Author
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Kato EE, Pimenta LA, de Almeida MES, Zambelli VO, Dos Santos MF, and Sampaio SC
- Abstract
Antitumor property of Crotoxin (CTX), the major toxin from Crotalus durissus terrificus snake venom, has been demonstrated in experimental animal models and clinical trials. However, the direct action of this toxin on the significant events involved in neovascularization, which are essential for tumor growth and survival, has not been confirmed. This study investigated the effects of CTX on the key parameters of neovascularization in two- and three-dimensional culture models. Murine endothelial cell lines derived from thymus hemangioma (t.End.1) were treated at different concentrations of CTX (6.25-200 nM). Endothelial cell proliferation, cell adhesion, and actin cytoskeletal dynamics on laminin (10 µg/ml), type I collagen (10 µg/ml), and fibronectin (3 µg/ml) were evaluated along with the endothelial cell migration and formation of capillary-like tubes in 3D Matrigel. CTX concentration of 50 nM inhibited tube formation on 3D Matrigel and impaired cell adhesion, proliferation, and migration under both culture medium and tumor-conditioned medium. These actions were not accountable for the loss of cell viability. Inhibition of cell adhesion to different extracellular matrix components was related to the reduction of αv and α2 integrin distribution and cytoskeletal actin polymerization (F-actin), accompanied by inhibition of focal adhesion kinase (FAK), Rac1 (GTPase) signaling proteins, and actin-related protein 2/3 (Arp 2/3) complex. This study proved that CTX inhibits the major events involved in angiogenesis, particularly against tumor stimuli, highlighting the importance of the anti-angiogenic action of CTX in inhibition of tumor progression., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kato, Pimenta, Almeida, Zambelli, Santos and Sampaio.)
- Published
- 2021
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44. Heat-Labile Toxin from Enterotoxigenic Escherichia coli Causes Systemic Impairment in Zebrafish Model.
- Author
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Henrique C, Falcão MAP, De Araújo Pimenta L, Maleski ALA, Lima C, Mitsunari T, Sampaio SC, Lopes-Ferreira M, and Piazza RMF
- Subjects
- Animals, Bacterial Toxins pharmacokinetics, Caco-2 Cells, Edema chemically induced, Embryo, Nonmammalian abnormalities, Embryo, Nonmammalian drug effects, Embryo, Nonmammalian metabolism, Enterotoxins pharmacokinetics, Escherichia coli Proteins pharmacokinetics, Heart Defects, Congenital chemically induced, Heart Rate drug effects, Humans, Intestines metabolism, Myocardium metabolism, Yolk Sac drug effects, Zebrafish abnormalities, Zebrafish metabolism, Bacterial Toxins toxicity, Enterotoxigenic Escherichia coli, Enterotoxins toxicity, Escherichia coli Proteins toxicity
- Abstract
Heat-labile toxin I (LT-I), produced by strains of enterotoxigenic Escherichia coli (ETEC), causes profuse watery diarrhea in humans. Different in vitro and in vivo models have already elucidated the mechanism of action of this toxin; however, their use does not always allow for more specific studies on how the LT-I toxin acts in systemic tracts and intestinal cell lines. In the present work, zebrafish ( Danio rerio ) and human intestinal cells (Caco-2) were used as models to study the toxin LT-I. Caco-2 cells were used, in the 62nd passage, at different cell concentrations. LT-I was conjugated to FITC to visualize its transport in cells, as well as microinjected into the caudal vein of zebrafish larvae, in order to investigate its effects on survival, systemic traffic, and morphological formation. The internalization of LT-I was visualized in 3 × 10
4 Caco-2 cells, being associated with the cell membrane and nucleus. The systemic traffic of LT-I in zebrafish larvae showed its presence in the cardiac cavity, yolk, and regions of the intestine, as demonstrated by cardiac edema (100%), the absence of a swimming bladder (100%), and yolk edema (80%), in addition to growth limitation in the larvae, compared to the control group. There was a reduction in heart rate during the assessment of larval survival kinetics, demonstrating the cardiotoxic effect of LT-I. Thus, in this study, we provide essential new depictions of the features of LT-I.- Published
- 2021
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45. Structure and species composition of diatom community during the wet season in three floodplain lakes of Brazilian Pantanal.
- Author
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Nardelli MS, Bicudo DC, Sampaio SC, and Cordovil CMDS
- Subjects
- Biodiversity, Brazil, Ecosystem, Environmental Monitoring methods, Lakes, Nitrogen chemistry, Phosphorus chemistry, Seasons, Diatoms physiology
- Abstract
In order to access environmental conditions, the use of bioindicators that have a close relationship with environmental stressors is a largely common practice, but when evaluating environmental inferences, the individual dominant taxa need to be interpreted. Humid regions such as the marshlands are fragile ecosystems and sustain communities of microalgae, often used as bioindicators, of which diatoms are a good example. Although they provide an excellent response to chemical and physical changes in water, diatom studies in surface sediments in wetlands are scarce worldwide. To determine whether diatom species have the potential to provide unambiguous inferences in the influence of environmental factors, we have evaluated diatom abundance in surface sediment, from three Pantanal lakes, against a set of environmental gradients: pH, dissolved oxygen, turbidity, conductivity, total dissolved solids, water temperature, index of trophic water status, total phosphorus and total nitrogen. The Ferradura lake presented an oligotrophic state and both Burro and Caracará lakes presented mesotrophic state. Diatoms were more abundant in the a mesotrophic conditions, but with higher species richness in the oligotrophic conditions. Depending on the N:P ratio, the nutrients nitrogen and phosphorus can also play the role of pollutants and may have negative and unpredictable effects in the environment, such as biotic homogenization. Despite the spatial variation in species, there was a greater richness of Eunotia Ehrenberg species, with the highest relative density of Eunotia formica Ehrenberg and E. pantropica Glushchenko, Kulikovskiy & Kociolek, due to the environmental acidic conditions, a determining characteristic of this genus. It was also observed that a small increase in the level of phosphorus generated an increase in the abundance of Aulacoseira Thwaites with the highest relative density of A. pusilla (Meister) Tuji & Houki and A. veraluciae Tremarin, Torgan & T.Ludwig. However, A. italica dominated in the moderately acidic environment. The results can help with decisions in impacted areas to solve socioeconomic problems, environmental management and biodiversity., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2021
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46. Physicochemical quality of and Escherichia coli resistance profiles in urban surface waters.
- Author
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Malagi I, Sampaio SC, Pinto FGS, Rosa DM, and Reis RRD
- Subjects
- Anti-Bacterial Agents, Bacteria, Rivers, Water Microbiology, Ecosystem, Escherichia coli
- Abstract
Aquatic ecosystems of urban rivers are contaminated through waste disposal, which poses a public health problem. The objective of this research was to evaluate the quality of water used for recreation and public supply of six rivers in the city of Cascavel - Paraná, including Cascavel, Quati, Bezerra, Antas, Clarito and Amambay. Samples were collected every 4 months in 2017, and their physicochemical and microbiological parameters, as well as resistance profiles of strains of Escherichia coli to antimicrobials distributed by pharmacies of the primary healthcare network, were evaluated. Parameters such as water temperature, turbidity, total nitrogen, total phosphorus, total coliforms and thermotolerant coliforms showed significant differences. The allowed limit for thermotolerant coliforms, which was set by National Environment Council, Resolution 357/2005, was exceeded in all of the six analyzed rivers. It was determined that 48.1% of E. coli strains showed resistance to nine antimicrobial tested. The highest levels of resistance were found for ampicillin (27.7%), tetracycline (27.7%) and amoxicillin (24.0%). The results of this study contribute to the understanding of the hazards associated with the contamination of springs in urban centers with wastewater containing resistant bacteria. Therefore, recovery work is necessary in these areas because of the importance of these water sources for the entire western region of Paraná state.
- Published
- 2020
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47. Metalloproteinases Suppression Driven by the Curcumin Analog DM-1 Modulates Invasion in BRAF-Resistant Melanomas.
- Author
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de Souza N, de Oliveira ÉA, Faião-Flores F, Pimenta LA, Quincoces JAP, Sampaio SC, and Maria-Engler SS
- Subjects
- Antineoplastic Agents chemical synthesis, Antineoplastic Agents chemistry, Cell Cycle Checkpoints drug effects, Cell Movement drug effects, Cell Proliferation drug effects, Cell Survival drug effects, Dose-Response Relationship, Drug, Drug Screening Assays, Antitumor, Humans, Melanoma metabolism, Melanoma pathology, Metalloproteases metabolism, Molecular Structure, Proto-Oncogene Proteins B-raf metabolism, Structure-Activity Relationship, Tumor Cells, Cultured, Antineoplastic Agents pharmacology, Drug Resistance, Neoplasm drug effects, Melanoma drug therapy, Metalloproteases antagonists & inhibitors, Proto-Oncogene Proteins B-raf antagonists & inhibitors
- Abstract
Background: Melanoma is the most aggressive skin cancer, and BRAF (V600E) is the most frequent mutation that led to the development of BRAF inhibitors (BRAFi). However, patients treated with BRAFi usually present recidivism after 6-9 months. Curcumin is a turmeric substance, and it has been deeply investigated due to its anti-inflammatory and antitumoral effects. Still, the low bioavailability and biodisponibility encouraged the investigation of different analogs. DM-1 is a curcumin analog and has shown an antitumoral impact in previous studies., Methods: Evaluated DM-1 stability and cytotoxic effects for BRAFi-sensitive and resistant melanomas, as well as the role in the metalloproteinases modulation., Results: DM-1 showed growth inhibitory potential for melanoma cells, demonstrated by reduction of colony formation, migration and endothelial tube formation, and cell cycle arrest. Subtoxic doses were able to downregulate important Metalloproteinases (MMPs) related to invasiveness, such as MMP-1, -2 and -9. Negative modulations of TIMP-2 and MMP-14 reduced MMP-2 and -9 activity; however, the reverse effect is seen when increased TIMP-2 and MMP-14 resulted in raised MMP-2., Conclusion: These findings provide essential details into the functional role of DM-1 in melanomas, encouraging further studies in the development of combinatorial treatments for melanomas., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)
- Published
- 2020
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48. Biochemical and biological characterization of the Hypanus americanus mucus: A perspective on stingray immunity and toxins.
- Author
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Coelho GR, Neto PP, Barbosa FC, Dos Santos RS, Brigatte P, Spencer PJ, Sampaio SC, D'Amélio F, Pimenta DC, and Sciani JM
- Subjects
- Animals, Brazil, Female, Immunity, Mucosal, Immunologic Techniques methods, Mucus immunology, Skates, Fish, Fish Venoms chemistry, Immunity, Innate, Immunologic Techniques veterinary, Mucus chemistry
- Abstract
Stingrays skin secretions are largely studied due to the human envenoming medical relevance of the sting puncture that evolves to inflammatory events, including necrosis. Such toxic effects can be correlated to the biochemical composition of the sting mucus, according to the literature. Fish skin plays important biological roles, such as the control of the osmotic pressure gradient, protection against mechanical forces and microorganism infections. The mucus, on the other hand, is a rich and complex fluid, acting on swimming, nutrition and the innate immune system. The elasmobranch's epidermis is a tissue composed mainly by mucus secretory cells, and marine stingrays have already been described to present secretory glands spread throughout the body. Little is known about the biochemical composition of the stingray mucus, but recent studies have corroborated the importance of mucus in the envenomation process. Aiming to assess the mucus composition, a new non-invasive mucus collection method was developed that focused on peptides and proteins, and biological assays were performed to analyze the toxic and immune activities of the Hypanus americanus mucus. Pathophysiological characterization showed the presence of peptidases on the mucus, as well as the induction of edema and leukocyte recruitment in mice. The fractionated mucus improved phagocytosis on macrophages and showed antimicrobial activity against T. rubrumç. neoformans and C. albicans in vitro. The proteomic analyses showed the presence of immune-related proteins like actin, histones, hemoglobin, and ribosomal proteins. This protein pattern is similar to those reported for other fish mucus and stingray venoms. This is the first report depicting the Hypanus stingray mucus composition, highlighting its biochemical composition and importance for the stingray immune system and the possible role on the envenomation process., (Copyright © 2019 Elsevier Ltd. All rights reserved.)
- Published
- 2019
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49. Advanced glycation endproducts produced by in vitro glycation of type I collagen modulate the functional and secretory behavior of dorsal root ganglion cells cultivated in two-dimensional system.
- Author
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Bufalo MC, Almeida ME, Franca IA, Zambelli VO, Martins Sant'anna MB, Kimura LF, Giardini AC, Cury Y, and Sampaio SC
- Subjects
- Actin Cytoskeleton metabolism, Actins metabolism, Animals, Cell Survival, Cells, Cultured, Enzyme Activation, Glycosylation, MAP Kinase Signaling System, Male, Mice, Nitrites metabolism, Phosphorylation, Rats, Wistar, Receptor for Advanced Glycation End Products metabolism, Tumor Necrosis Factor-alpha biosynthesis, Cell Culture Techniques methods, Collagen Type I metabolism, Ganglia, Spinal metabolism, Glycation End Products, Advanced metabolism
- Abstract
Advanced glycation end-products (AGEs) are proteins/lipids that are glycated upon sugar exposure and are often increased during inflammatory diseases such as osteoarthritis and neurodegenerative disorders. Here, we developed an extracellular matrix (ECM) using glycated type I collagen (ECM-GC), which produced similar levels of AGEs to those detected in the sera of arthritic mice. In order to determine whether AGEs were sufficient to stimulate sensory neurons, dorsal root ganglia (DRGs) cells were cultured on ECM-GC or ECM-NC-coated plates. ECM-GC or ECM-NC were favorable for DRG cells expansion. However, ECM-GC cultivated neurons displayed thinner F-actin filaments, rounded morphology, and reduced neuron interconnection compared to ECM-NC. In addition, ECM-GC did not affect RAGE expression levels in the neurons, although induced rapid p38, MAPK and ERK activation. Finally, ECM-GC stimulated the secretion of nitrite and TNF-α by DRG cells. Taken together, our in vitro glycated ECM model suitably mimics the in vivo microenvironment of inflammatory disorders and provides new insights into the role of ECM impairment as a nociceptive stimulus., (Copyright © 2019. Published by Elsevier Inc.)
- Published
- 2019
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50. QSPR modelling of the soil sorption coefficient from training sets of different sizes.
- Author
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Olguin CJM, Sampaio SC, Dos Reis RR, Remor MB, and Olguin CFA
- Subjects
- Adsorption, Data Interpretation, Statistical, Organic Chemicals chemistry, Quantitative Structure-Activity Relationship, Reproducibility of Results, Risk Assessment, Soil Pollutants chemistry, Models, Chemical, Soil chemistry
- Abstract
Quantitative structure-property relationship (QSPR) modelling has been used in many scientific fields. This approach has been extensively applied in environmental research to predict physicochemical properties of compounds with potential environmental impact. The soil sorption coefficient is an important parameter for the evaluation of environmental risks, and it helps to determine the final fate of substances in the environment. In the last few years, different QSPR models have been developed for the determination of the sorption coefficient. In this study, several QSPR models were generated and evaluated for the prediction of log K
oc from the relationship with log P. These models were obtained from an extensive and diverse training set (n = 639) and from subsets of this initial set (i.e. halves, fourths and eighths). The aim of this study was to investigate whether the size of the training set affects the statistical quality of the obtained models. Furthermore, statistical equivalence was verified between the models obtained from smaller sets and the model obtained from the total training set. The results confirmed the equivalence between the models, thus indicating the possibility of using smaller training sets without compromising the statistical quality and predictive capability, as long as most chemical classes in the test set are represented in the training set.- Published
- 2019
- Full Text
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