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1.
Transfusion ; 64(3): 501-509, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38258881

RESUMO

BACKGROUND: Transfusion-transmitted malaria (TTM) is a public health problem in endemic and nonendemic areas. The Brazilian Ministry of Health (MH) requested the development of a nucleic acid amplification test (NAT) for the detection of Plasmodium spp. in public blood centers to increase blood safety. STUDY DESIGN AND METHODS: The new Brazilian NAT kit named NAT PLUS HIV/HBV/HCV/Malaria Bio-Manguinhos was first implemented in HEMORIO, a public blood center in the city of Rio de Janeiro. Since October 1, 2022, this blood center has been testing all its blood donations for malaria in a pool of six plasma samples to detect Plasmodium spp. by real-time polymerase chain reaction (PCR). RESULTS: Since the implementation of the NAT PLUS platform until February 2023, HEMORIO has successfully received and tested 200,277 donations. The platform detected two asymptomatic donors in the city of Rio de Janeiro, which is a nonendemic region for malaria. Our analyses suggested a malaria from the Amazon region caused by Plasmodium vivax, in the first case, while an autochthonous transmission case by Plasmodium malariae was identified in the rural area of Rio de Janeiro state. DISCUSSION: The NAT PLUS platform detects Plasmodium spp. in plasma samples with sensitivity capable of detecting subpatent infections. This is the first time worldwide that a group developed and implemented molecular diagnosis for Plasmodium spp. to be used by public blood centers to avoid TTM.


Assuntos
Infecções por HIV , Hepatite C , Malária , Humanos , Vírus da Hepatite B , Doadores de Sangue , Brasil/epidemiologia , Malária/diagnóstico , Malária/epidemiologia , Plasmodium malariae , Infecções por HIV/diagnóstico , Infecções por HIV/epidemiologia
2.
Mem Inst Oswaldo Cruz ; 118: e220202, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36946838

RESUMO

BACKGROUND: The prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) has changed unevenly over time around the world. Although whole genome sequencing is the gold standard for virus characterisation, the discovery of alpha VOC causing spike gene target failure (SGTF) result, when tested using an reverse transcription real-time polymerase chain reaction (RT-qPCR) assay, has provided a simple tool for tracking the frequencies of variants. OBJECTIVES: The aim of this study was to investigate if a multiplex RT-qPCR assay (BioM 4Plex VOC) could be used to detect SARS-CoV-2 and to perform a VOC screening test in a single reaction tube. Here, we present the multicentre study evaluating this assay. METHODS: Twelve laboratories have participated in the multicentre study. The BioM 4Plex VOC was distributed to them with detailed instructions of how to perform the test. They were asked to test the BioM 4Plex VOC in parallel with their routine Commercial SARS-CoV-2 diagnostic assay. Additionally, they were requested to select SARS-CoV-2-positive samples with genome sequenced and lineage definition according to PANGO lineage classification. FINDINGS: The BioM 4Plex VOC and commercial RT-PCR assay are equally effective in detecting SARS-CoV-2. Results revealed a specificity of 96.5-100% [95% confidence interval (CI)], a sensitivity of 99.8-100% (95% CI), and an accuracy of 99.8-100% (95% CI). A 99% concordance rate was found between results from the BioM 4Plex VOC and that from available genome sequencing data. MAIN CONCLUSIONS: The BioM 4Plex VOC provides an effective solution to detect SARS-CoV-2 infections and screening for VOCs in a single reaction. It is a straightforward method to help us monitor the frequency and distribution of VOCs and develop strategies to better cope with the pandemics.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , COVID-19/diagnóstico , Teste para COVID-19 , Bioensaio , Mapeamento Cromossômico
3.
Malar J ; 19(1): 275, 2020 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-32736625

RESUMO

BACKGROUND: Malaria can be transmitted by blood transfusion through donations collected from asymptomatic donors. Transfusion-transmitted malaria (TTM) poses a great risk to blood services worldwide. A good screening tool for Plasmodium spp. detection in blood banks must have a high sensitivity for prevention of TTM. However, in Brazilian blood banks, screening for malaria still relies on microscopy. METHODS: In Brazil, screening for human immunodeficiency virus type 1 (HIV), RNA/DNA for hepatitis C (HCV) and hepatitis B (HBV) viruses is mandatory for every blood donation and uses nucleic acid amplification testing (NAT). The aim of this study was to evaluate the inclusion of an assay for malaria to identify Plasmodium sp. from total nucleic acid (TNA; DNA/RNA) by targeting the 18S rRNA gene of the parasite. RESULTS: Considering the limitations of microscopy and the wide availability of the Brazilian NAT platform in the screening of blood units for HIV, HCV, and HBV, a molecular diagnostic tool was validated for detection of Plasmodium sp. in blood banks; a pilot study showed that using this novel NAT assay could reduce the risk of TTM. CONCLUSION: The prototype HIV/HCV/HBV/malaria NAT assay was effective in detecting infected candidate donors and has good prospects to be applied in routine screening for preventing TTM.


Assuntos
Malária/prevenção & controle , Programas de Rastreamento/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Plasmodium/isolamento & purificação , Vigilância da População/métodos , Adolescente , Adulto , Bancos de Sangue , Transfusão de Sangue , Brasil , Monitoramento Epidemiológico , Feminino , Infecções por HIV/diagnóstico , Hepatite B/diagnóstico , Hepatite C/diagnóstico , Humanos , Malária/transmissão , Masculino , Programas de Rastreamento/instrumentação , Pessoa de Meia-Idade , Projetos Piloto , Plasmodium/genética , RNA de Protozoário/análise , RNA Ribossômico 18S/análise , Adulto Jovem
5.
Genet Mol Biol ; 43(1): e20180237, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31644681

RESUMO

Hepatitis C virus (HCV) infection is a worldwide health problem. Nowadays, direct-acting antiviral agents (DAAs) are the main treatment for HCV; however, the high level of virus variability leads to the development of resistance-associated variants (RAVs). Thus, assessing RAVs in infected patients is important for monitoring treatment efficacy. The aim of our study was to investigate the presence of naturally occurring resistance mutations in HCV NS3 and NS5 regions in treatment-naïve patients. Ninety-six anti-HCV positive serum samples from blood donors at the Center of Hematology and Hemotherapy of Santa Catarina State (HEMOSC) were collected retrospectively in 2013 and evaluated in this study. HCV 1a (37.9%), 1b (25.3%), and 3a (36.8%) subtypes were found. The frequency of patients with RAVs in our study was 6.9%. The HCV NS5b sequencing reveled 1 sample with L320F mutation and 4 samples with the C316N/R polymorphism. The analysis of the NS3 region revealed the D168A/G/T (3.45%), S122G (1.15%), and V55A (2.3%) mutations. All samples from genotype 3a (36.8%) presented the V170 I/V non-synonymous mutation. In conclusion, we have shown that mutations in NS3 and NS5b genes are present in Brazilian isolates from therapy-naïve HCV patients.

6.
Transfusion ; 58(4): 862-870, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29383720

RESUMO

BACKGROUND: The history of the development and implementation of the Brazilian nucleic acid testing (NAT) platform to detect and discriminate among human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B virus (HBV) infections in blood donors is described here. The results for the sensitivity, reproducibility, and NAT yield of the platform since program implementation are provided. STUDY DESIGN AND METHODS: The Brazilian NAT HIV, HCV, and HBV kit was developed and evaluated with regard to analytical sensitivity, specificity, intralot and interlot reproducibility, interfering substances, and genotype and diagnostic sensitivity. Additionally, a sample of identified NAT-yield cases was characterized with regard to viral load. RESULTS: The 95% limits of detection for HIV, HCV, and HBV were 68.02, 102.35, and 9.08 IU/mL, respectively. All replicates were detected with reproducibility assays between the acceptable values. A total of 13,610,536 blood donors was screened from 2010 to 2016, and 63 HIV-yield cases and 28 HCV-yield cases were detected. Among 5,795,424 blood donors screened for HBV from 2014 to 2016, 42 yield cases were found. CONCLUSION: The Brazilian NAT HIV, HCV, and HBV kit is an automated NAT system suitable for routine blood donor screening in a completely traceable process. The analytical sensitivity as well as the diagnostic sensitivity fulfilled all requirements set by the health ministry for blood donor screening. A significant number of transmission cases were prevented by the implementation of this important program.


Assuntos
Doadores de Sangue , Segurança do Sangue , DNA Viral/sangue , Infecções por HIV/diagnóstico , Hepatite B/diagnóstico , Hepatite C/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real , Viremia/diagnóstico , Automação , Segurança do Sangue/instrumentação , Segurança do Sangue/métodos , Segurança do Sangue/normas , Brasil , Infecções por HIV/sangue , Hepatite B/sangue , Hepatite C/sangue , Humanos , Programas de Rastreamento/métodos , Reação em Cadeia da Polimerase em Tempo Real/instrumentação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase em Tempo Real/normas , Padrões de Referência , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Carga Viral , Viremia/transmissão
7.
Reprod Fertil Dev ; 30(10): 1314-1328, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29681258

RESUMO

Mammalian oocytes resume meiosis spontaneously after removal from the ovarian follicle. We tested the effects of a 2-h prematuration treatment (Pre-IVM) with forskolin (FSK) and 3-isobutyl-1-methylxanthine (IBMX) in bovine cumulus-oocyte complexes (COCs) on the lipid content of oocytes and blastocysts, on the membrane lipid composition of blastocysts and on the transcriptional profiling of cumulus cells and blastocysts in a high-throughput platform. Embryonic development rates to the morula (mean 56.1%) or blastocyst (mean 26.3%) stages were unaffected by treatment. Lipid content was not affected after Pre-IVM, but was increased after IVM in treated oocytes. Conversely, the lipid content was reduced in Pre-IVM blastocysts. Pre-IVM COCs generated blastocysts containing blastomeres with more unsaturated lipids in their membranes. Pre-IVM also altered the relative abundance of 31 gene transcripts after 2h and 16 transcripts after 24h in cumulus cells, while seven transcripts were altered in blastocysts. Our results suggest that the Pre-IVM treatment affected the lipid composition and transcriptional profiles of COCs and blastocysts. Therefore, Pre-IVM with FSK and IBMX could be used either to prevent spontaneous meiotic resumption during IVM or to modulate lipid composition in the membrane and cytoplasm of blastocysts, potentially improving bovine embryos.


Assuntos
Blastocisto/efeitos dos fármacos , Blastocisto/metabolismo , Células do Cúmulo/efeitos dos fármacos , Células do Cúmulo/metabolismo , AMP Cíclico/agonistas , AMP Cíclico/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , 1-Metil-3-Isobutilxantina/farmacologia , Animais , Blastocisto/citologia , Bovinos , Colforsina/farmacologia , Células do Cúmulo/citologia , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/genética , Feminino , Técnicas de Maturação in Vitro de Oócitos , Oócitos/crescimento & desenvolvimento , Gravidez , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transcriptoma/efeitos dos fármacos
8.
Anal Bioanal Chem ; 409(29): 6803-6812, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28948318

RESUMO

Banana is one of the most popular fruits in the world but has been substantially impaired by Panama disease in the last years. Fusarium oxysporum f. sp. cubense (Foc) is the causal agent and colonizes banana cultivars from many subgroups with different aggressiveness levels, often leading to plant death while compromising new crops in infested areas. This study has evaluated the ability of MALDI-MS protein and lipid fingerprinting to provide intraspecies classification of Foc isolates and to screen biomolecules related to host-pathogen relationship. The MS data, when inspected via partial least square discriminant analysis (PLS-DA), distinguished the isolates by aggressiveness as well as by specific location and host. Although both lipids and proteins show discriminating tendencies, these differences were more clearly perceived via the protein profiles. Considering that Cavendish cultivar is the more resistant option to endure Foc presence in the field, the lipids and proteins related to this subgroup might have an important role in pathogen adaptation. This study reports a new application of MALDI-MS for the analysis of a banana pathogen with intraspecies classification ability. Graphical abstract MALDI-MS classified Foc isolates by aggressiveness level on banana revealing the additional influence of location and host cultivar on the expression of lipids and proteins.


Assuntos
Proteínas Fúngicas/química , Fusarium/química , Fusarium/classificação , Lipídeos/química , Mapeamento de Peptídeos , Doenças das Plantas/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
9.
Korean J Parasitol ; 55(3): 239-246, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28719948

RESUMO

Strains of Toxoplasma gondii in Brazil are highly genetically diverse compared to strains from North America and Europe. Dogs are epidemiologically important because they act as sentinels for T. gondii infections in humans and are good indicators of environmental contamination. The aim of this study was to isolate and genetically characterize T. gondii strains from tissues of naturally infected Brazilian dogs. For this study, 21 blood samples were collected from dogs at the Zoonosis Control Centers of Ilhéus and Itabuna cities, Bahia, Brazil. The sera were examined for T. gondii antibodies using the indirect hemagglutination test. Brains and hearts of seropositive dogs were bioassayed in mice to isolate and characterize T. gondii parasites by PCR-RFLP using 10 genetic markers (SAG1, newSAG2, SAG3, BTUB, c22-8, c29-2, GRA6, PK1, APICO, and L358). However, T. gondii was isolated from only 4 (57.1%) dogs, designated TgDgBr6, 13, 17, and 21. All strains were virulent, causing clinical changes (rough hair coat, lethargy, and abdominal distention) and the death of all mice within 8-20 days after inoculation. Genetic analysis of these 4 T. gondii isolates revealed 4 distinct genotypes with different clonal lineage combinations (types I, II, and III) and 2 atypical alleles. Using PCR-RFLP with several markers, this study contributes to evaluations of the genetic diversity of strains circulating in Brazil.


Assuntos
Doenças do Cão/parasitologia , Cães/parasitologia , Genótipo , Toxoplasma/genética , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/parasitologia , Alelos , Animais , Anticorpos Antiprotozoários/sangue , Encéfalo/parasitologia , Brasil , Variação Genética , Técnicas de Genotipagem , Coração/parasitologia , Humanos , Camundongos , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Toxoplasma/imunologia , Toxoplasma/patogenicidade , Virulência
10.
J Virol ; 89(23): 11871-83, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26378175

RESUMO

UNLABELLED: Dengue is one of the main public health concerns worldwide. Recent estimates indicate that over 390 million people are infected annually with the dengue virus (DENV), resulting in thousands of deaths. Among the DENV nonstructural proteins, the NS1 protein is the only one whose function during replication is still unknown. NS1 is a 46- to 55-kDa glycoprotein commonly found as both a membrane-associated homodimer and a soluble hexameric barrel-shaped lipoprotein. Despite its role in the pathogenic process, NS1 is essential for proper RNA accumulation and virus production. In the present study, we identified that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) interacts with intracellular NS1. Molecular docking revealed that this interaction occurs through the hydrophobic protrusion of NS1 and the hydrophobic residues located at the opposite side of the catalytic site. Moreover, addition of purified recombinant NS1 enhanced the glycolytic activity of GAPDH in vitro. Interestingly, we observed that DENV infection promoted the relocalization of GAPDH to the perinuclear region, where NS1 is commonly found. Both DENV infection and expression of NS1 itself resulted in increased GAPDH activity. Our findings indicate that the NS1 protein acts to increase glycolytic flux and, consequently, energy production, which is consistent with the recent finding that DENV induces and requires glycolysis for proper replication. This is the first report to propose that NS1 is an important modulator of cellular energy metabolism. The data presented here provide new insights that may be useful for further drug design and the development of alternative antiviral therapies against DENV. IMPORTANCE: Dengue represents a serious public health problem worldwide and is caused by infection with dengue virus (DENV). Estimates indicate that half of the global population is at risk of infection, with almost 400 million cases occurring per year. The NS1 glycoprotein is found in both the intracellular and the extracellular milieus. Despite the fact that NS1 has been commonly associated with DENV pathogenesis, it plays a pivotal but unknown role in the replication process. In an effort to understand the role of intracellular NS1, we demonstrate that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) interacts with NS1. Our results indicate that NS1 increases the glycolytic activity of GAPDH in vitro. Interestingly, the GAPDH activity was increased during DENV infection, and NS1 expression alone was sufficient to enhance intracellular GAPDH activity in BHK-21 cells. Overall, our findings suggest that NS1 is an important modulator of cellular energy metabolism by increasing glycolytic flux.


Assuntos
Metabolismo Energético/fisiologia , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora)/metabolismo , Modelos Moleculares , Proteínas não Estruturais Virais/química , Proteínas não Estruturais Virais/metabolismo , Análise de Variância , Animais , Linhagem Celular , Cromatografia Líquida , Cricetinae , Ensaio de Imunoadsorção Enzimática , Células Endoteliais da Veia Umbilical Humana , Humanos , Imunoprecipitação , Microscopia de Fluorescência , Ligação Proteica , Espectrometria de Massas em Tandem , Proteínas não Estruturais Virais/genética
11.
Rev Bras Parasitol Vet ; 33(1): e020223, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38511819

RESUMO

This study aimed to detect Toxoplasma gondii in artisanal salted meat products sold in street markets in the Ilhéus-Itabuna microregion and to assess the salt concentration used in their preparation and its influence on the parasite's viability. A total of 125 samples of various artisanal meat products sold in street markets located in the Ilhéus-Itabuna microregion were collected during 2021. Serological analysis using indirect hemagglutination (HAI) and molecular analysis (PCR) were performed on these samples to detect the presence of the parasite. Möhr's method was utilized to determine the sodium chloride concentration in the samples. Of all samples, 21 were subjected to a bioassay in albino mice to verify the viability of possible tissue cysts. Among the 125 meat products, 10 (8%) tested positive in the serological analysis including four cured pork sausages, five beef sun-dried meats, and one mixed fresh sausage (pork and chicken). None of 125 samples tested positive in the molecular analysis. On bioassay, all mice tested negative for the presence of the parasite. The NaCl concentration in the positive samples ranged from 2.9% to 8%. The results demonstrated that the salt concentration in the collected samples was sufficient to inactivate the parasite T. gondii.


Assuntos
Doenças dos Bovinos , Produtos da Carne , Doenças dos Roedores , Toxoplasma , Toxoplasmose Animal , Bovinos , Animais , Camundongos , Produtos da Carne/parasitologia , Cloreto de Sódio , Carne/parasitologia , Bioensaio/veterinária
12.
J Nat Prod ; 76(9): 1559-64, 2013 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-23978079

RESUMO

Four species of gonyleptid harvestmen, Acanthogonyleptes pulcher, Gonyleptes saprophilus (Gonyleptinae), Sodreana barbiellini, and Sodreana leprevosti (Sodreaninae), were examined by GC-MS and ¹³H NMR. All of these species release vinyl ketones, and three of them produce the corresponding pyranyl ketones, which are presumed hetero-Diels-Alder (HDA) dimers. The vinyl ketones 5-methyl-1-hexen-3-one, rac-4-methyl-1-hexen-3-one, and (S)-4-methyl-1-hexen-3-one were synthesized. Natural 4-methyl-1-hexen-3-one is present as a single stereoisomer and has the R-configuration. Vinyl ketone dimers (HDA dimers) were also observed in the scent gland exudate and characterized by HRMS, ¹³C NMR, and ¹H NMR chemical shifts of the pyranyl moiety.


Assuntos
Aracnídeos/química , Cetonas/química , Piranos/química , Glândulas Odoríferas/química , Compostos de Vinila/química , Animais , Aracnídeos/classificação , Aracnídeos/genética , Cromatografia Gasosa-Espectrometria de Massas , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Glândulas Odoríferas/metabolismo , Estereoisomerismo
13.
Molecules ; 18(9): 11429-51, 2013 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-24043140

RESUMO

Benzoquinones are usually present in arthropod defence exudates. Here, we describe the chemical profiles of 12 harvestman species belonging to the neotropical family Gonyleptidae. Nine of the studied species produced benzoquinones, while three produced alkyl phenols. Two benzoquinones and one phenol exhibited biological activity against bacteria and fungi. We also studied the biosynthesis of 2-ethyl-1,4-benzoquinone by feeding Magnispina neptunus individuals with ¹³C-labelled precursors; the benzoquinones were biosynthesised through a polyketide pathway using acetate and propionate building blocks.


Assuntos
Aracnídeos/metabolismo , Benzoquinonas/química , Fenóis/química , Animais , Antibacterianos/química , Antibacterianos/metabolismo , Antibacterianos/farmacologia , Antifúngicos/química , Antifúngicos/metabolismo , Antifúngicos/farmacologia , Aracnídeos/química , Bacillus/efeitos dos fármacos , Benzoquinonas/metabolismo , Benzoquinonas/farmacologia , Vias Biossintéticas , Candida albicans/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Micrococcus luteus/efeitos dos fármacos , Estrutura Molecular , Fenóis/metabolismo , Fenóis/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos
14.
Viruses ; 15(11)2023 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-38005909

RESUMO

Chikungunya fever, a debilitating disease caused by Chikungunya virus (CHIKV), is characterized by a high fever of sudden onset and an intense arthralgia that impairs individual regular activities. Although most symptoms are self-limited, long-term persistent arthralgia is observed in 30-40% of infected individuals. Currently, there is no vaccine or specific treatment against CHIKV infection, so there is an urgent need for the discovery of new therapeutic options for CHIKF chronic cases. This present study aims to test the antiviral, cytoprotective, and anti-inflammatory activities of an ethanol extract (FF72) from Ampelozizyphus amazonicus Ducke wood, chemically characterized using mass spectrometry, which indicated the major presence of dammarane-type triterpenoid saponins. The major saponin in the extract, with a deprotonated molecule ion m/z 897 [M-H]-, was tentatively assigned as a jujubogenin triglycoside, a dammarane-type triterpenoid saponin. Treatment with FF72 resulted in a significant reduction in both virus replication and the production of infective virions in BHK-21-infected cells. The viability of infected cells was assessed using an MTT, and the result indicated that FF72 treatment was able to revert the toxicity mediated by CHIKV infection. In addition, FF72 had a direct effect on CHIKV, since the infectivity was completely abolished in the presence of the extract. FF72 treatment also reduced the expression of the major pro-inflammatory mediators overexpressed during CHIKV infection, such as IL-1ß, IL-6, IL-8, and MCP-1. Overall, the present study elucidates the potential of FF72 to become a promising candidate of herbal medicine for alphaviruses infections.


Assuntos
Febre de Chikungunya , Vírus Chikungunya , Saponinas , Triterpenos , Humanos , Febre de Chikungunya/tratamento farmacológico , Madeira , Triterpenos/farmacologia , Replicação Viral , Saponinas/farmacologia , Antivirais/farmacologia , Antivirais/uso terapêutico , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Etanol/farmacologia , Artralgia/tratamento farmacológico , Damaranos
15.
Parasitol Res ; 110(2): 509-14, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21739310

RESUMO

This study aimed to compare the sensitivity of five diagnostic methods commonly used for the detection of Toxoplasma gondii in tissues of naturally infected pigs. We purchased 20 heads of pigs in butcher shops in the city of Ilhéus, Bahia. Brain and tongue fragments were taken from each animal for the performance of PCR against T. gondii. The rest of these two tissues were processed and inoculated into three mice. These rodents were observed for 42 days and euthanized. We prepared slides with brain and lungs of each mouse for the visualization of T. gondii. From the tissues of mice, we carried out polymerase chain reaction (PCR), histopathology, and immunohistochemistry in an attempt to identify the parasite. The PCR direct from the tissue of pigs showed 10% (2/20) of positive samples, all from the brain. PCR in tissue from mice found that 55% (11/20) of pigs were positive: 55% (11/20) and 45% (9/20) for brains and tongues, respectively. Mice were inoculated with material obtained from the samples and examined by various methods for resulting Toxoplasma infection (bioassay). Cyst detection in bioassay mice identified 25% (5/20) and immunohistochemistry 30% (6/20) of the samples pigs as positive for T. gondii. Histopathology of mice tissue could not detect parasite; only suggestive pathological changes such as inflammation with foci of necrosis were seen. The results indicated PCR of mice tissue as the most sensitive among those tested.


Assuntos
Técnicas de Laboratório Clínico/métodos , Parasitologia/métodos , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/parasitologia , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/diagnóstico , Medicina Veterinária/métodos , Experimentação Animal , Animais , Encéfalo/parasitologia , Histocitoquímica/métodos , Imuno-Histoquímica/métodos , Pulmão/parasitologia , Camundongos , Reação em Cadeia da Polimerase/métodos , Sensibilidade e Especificidade , Suínos , Língua/parasitologia
16.
Artif Cells Nanomed Biotechnol ; 50(1): 158-167, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35713365

RESUMO

The aim of this study was to evaluate the effect of multilamellar vesicles (MLVs) of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) in co-culture with in vitro-produced bovine embryos (IVPEs). The stability of five concentrations of MLVs (1.0, 1.25, 1.5, 1.75, and 2.0 mM) produced using ultrapure water or embryonic culture medium with 24 or 48 h of incubation at 38.5 °C with 5% CO2 was assessed. In addition, the toxicity of MLVs and their modulation of the lipid profile of the plasma membrane of IVPEs were evaluated after 48 h of co-culture. Both media allowed the production of MLVs. Incubation (24 and 48 h) did not impair the MLV structure but affected the average diameter. The rate of blastocyst production was not reduced, demonstrating the nontoxicity of the MLVs even at 2.0 mmol/L. The lipid profile of the embryos was different depending on the MLV concentration. In comparison with control embryos, embryos cultured with MLVs at 2.0 mmol/L had a higher relative abundance of six lipid ions (m/z 720.6, 754.9, 759.0, 779.1, 781.2, and 797.3). This study sheds light on a new culture system in which the MLV concentration could change the lipid profile of the embryonic cell membrane in a dose-dependent manner.


Assuntos
Blastocisto , Bicamadas Lipídicas , Animais , Bovinos , Membrana Celular , Bicamadas Lipídicas/química
17.
Nat Prod Res ; 36(13): 3450-3454, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33287568

RESUMO

This work aimed to carry out phytochemical prospecting and evaluate the antioxidant potential of Diplopterys pubipetala, a species of Malpighiaceae family that has not yet been studied.In qualitative analyses of hydroethanolic extracts of leaves and stems were detected the presence of flavonoids, alkaloidsand terpenes. The histochemical evaluation evidenced a greater distribution of these compounds in the tissues of leaf when compared with those of stem. The analysis by mass spectrometry allowed the identification of prenylated xanthones and glycoside flavonoids that have not yet been reported in the literature. The antioxidant activity of the stem extract was considered moderate (IAA = 0.79), but the leaves presented a strong antioxidant activity (IAA = 1.6). In this work we present information about the phytochemicals of D. pubipetala, showing that the species is promising in obtaining compounds with medicinal potential mainly antioxidant potential.


Assuntos
Malpighiaceae , Antioxidantes/química , Flavonoides/química , Malpighiaceae/química , Compostos Fitoquímicos/análise , Extratos Vegetais/química , Folhas de Planta/química
18.
J Clin Virol ; 146: 105054, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34920373

RESUMO

BACKGROUND: Chikungunya virus (CHIKV) causes a febrile syndrome with intense and debilitating arthralgia that can persist for several months or years after complete virus clearance. As there is no specific antiviral treatment or vaccine against CHIKV, identification of serological markers that help clinical management of CHIKV patients is urgent. The High Mobility Group Box 1 (HMGB1) protein is secreted to extracellular milieu and triggers an intense inflammatory process by inducing the overexpression of pro-inflammatory cytokines. HMGB1 plays an important role in several virus diseases as well as in rheumatoid arthritis. OBJECTIVES: This study focus on the investigation of HMGB1 serum levels in a sera panel from CHIKV-infected patients in an attempt to assess its potential as a biomarker for chikungunya clinical management. STUDY DESIGN: Eighty CHIKV-positive samples and 32 samples from healthy donors were subjected to a quantitative HMGB1 ELISA assay to assess the HMGB1 circulating levels. RESULTS: HMGB1 levels were significantly higher in CHIKV-positive samples (516.12 ng/mL, SEM ± 48.83 ng/mL) compared to negative control (31.20 ng/mL, SEM ± 3.24 ng/mL, p < 0.0001). Circulating levels of HMGB1 persisted elevated during the whole acute-phase of disease and correlated with virus titer (p < 0.05). CONCLUSIONS: The present study is the first to describe increased serum levels of HMGB1 in CHIKV infection and its positive correlation with virus titer, suggesting its potential use as a biomarker for diagnosis and treatment of chikungunya fever.


Assuntos
Febre de Chikungunya , Vírus Chikungunya , Proteína HMGB1 , Biomarcadores , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Proteína HMGB1/sangue , Humanos
19.
J Nat Prod ; 74(4): 658-63, 2011 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-21361365

RESUMO

The defensive secretions of five neotropical species of harvestmen (Opiliones: Gonyleptidae) from the Brazilian Atlantic Forest were analyzed and chemically characterized by GC-MS and NMR methods. Three of the species, Cobania picea, Roweria virescens, and Serracutisoma proximum, secrete a mixture of 2,3-dimethyl-1,4-benzoquinone and 2-ethyl-3-methyl-1,4-benzoquinone. The secretions produced by the other two species, Iporangaia pustulosa and Neosadocus maximus, contain 1-hepten-3-one, 5-methyl-1-hexen-3-one, and 1-(6-butyl-3,4-dihydro-2H-pyran-2-yl)pentanone (1) as major components, as well as 2,3-dimethyl-1,4-benzoquinone and 2-ethyl-3-methyl-1,4-benzoquinone as minor constituents. The dihydropyran 1-(6-butyl-3,4-dihydro-2H-pyran-2-yl)pentanone (1) is a new natural product, composed of two 1-hepten-3-one subunits formally linked in a hetero-Diels-Alder reaction. The natural product was proven to be racemic, and its biogenetic origin is discussed.


Assuntos
Aracnídeos/química , Piranos/isolamento & purificação , Animais , Brasil , Cromatografia Gasosa-Espectrometria de Massas , Masculino , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Pentanonas , Piranos/química , Piranos/farmacologia , Glândulas Odoríferas/metabolismo , Estereoisomerismo
20.
Asian J Transfus Sci ; 15(1): 82-86, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34349462

RESUMO

BACKGROUND: Brazilian blood banks encourage donors to report postdonation information (PDI) regarding conditions that would lead to deferral in an attempt to retrieve distributed nonconforming blood. OBJECTIVES: This study evaluated the profile of donors reporting PDI, the impact on transfusion safety, and the possible impact on the discard of blood products. SUBJECTS AND METHODS: We analyzed 115 consecutive PDIs between May 2014 and July 2015, a period comprising two dengue-like syndrome (DLS) outbreaks. RESULTS: These PDIs accounted for 87,780 blood donations. The average time for PDIs since donation was 4 (0-23) days and 190 blood components were discarded. DLS accounted for 21.7% of the PDIs analyzed; 11 of the 23 samples tested were nucleic acid test (NAT) positive for dengue and 2 positive for Zika virus (ZIKV). Six of these PDIs were reported after blood components have been transfused: After NAT testing, one of these recipients was diagnosed with dengue and another one with ZIKV infection, both possible transfusions transmitted but without clinical consequences. CONCLUSION: The high number of recovered blood components due to PDI suggests that PDI remains a great ally in the fight against transfusion-transmitted infections and may be particularly useful during outbreaks of emerging potentially blood-borne pathogens.

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