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1.
Isr Med Assoc J ; 21(7): 504, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31507133

RESUMO

BACKGROUND: Klotho is a transmembrane protein that can be shed and can act as a circulating hormone in three forms: soluble klotho (KL1 + KL2), KL1, and KL2. Klotho was discovered as a gene implicated in aging through inhibition of the IGF-I pathway. Our laboratory discovered the role of klotho as a tumor suppressor in breast cancer and other malignancies. Furthermore, we showed that the KL1 domain mediates this activity. Altered cancer cell metabolism is a hallmark of cancer and our lab demonstrated various effects of klotho on breast cancer cell metabolism. Thus, klotho inhibited glycolysis and activated adenosine monophosphate activating kinase (AMPK), an energy sensor pathway. Moreover, inhibition of AMPK reduced the tumor suppressor activity of klotho. OBJECTIVES: To assess the effect of KL1 on breast tumor cells metabolism, as KL1 possesses the tumor suppressor activity of klotho. METHODS: We used MCF-7 breast cancer cells treated with soluble or over-expressed KL1 and klotho. Glycolysis was assessed by measuring mRNA levels of key glycolytic enzymes using reverse transcription polymerase chain reaction and by measuring lactate and glucose levels in media. The AMPK pathway was studied by monitoring AMPK phosphorylation as well as its down-stream target, acetyl-CoA carboxylase, using western blotting. Wound healing assay was used to assess cell migration. RESULTS: KL1 treatment reduced glycolytic enzymes mRNA levels and the activity of hexokinase, similar to klotho treatment. Furthermore, KL1 reduced glucose uptake and decreased lactate production. KL1 elevated phosphorylated acetyl-CoA carboxylase and phosphorylated AMPK levels. Inhibition AMPK (using a mutant AMPK activator) stopped KL1 from inhibiting cell migration, suggesting AMPK underlies klotho's tumor suppressor activity. CONCLUSIONS: Our data indicate KL1 as a regulator of metabolic activity in breast cancer and suggest that metabolic alterations underlie KL1 tumor suppressor activities. Furthermore, as KL1 and klotho share a similar effect on cell metabolism, our results further support the central role KL1 domain plays in klotho's tumor suppressor activity.


Assuntos
Neoplasias da Mama/metabolismo , Glucuronidase/metabolismo , Glicólise/fisiologia , Proteínas Quinases Ativadas por AMP/metabolismo , Movimento Celular/fisiologia , Feminino , Humanos , Células MCF-7 , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
3.
Medicine (Baltimore) ; 98(36): e16705, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31490362

RESUMO

Deregulation of miR-153 has recently been observed in several common human cancer, while miR-153 serves an oncogene or tumor suppressive role in different cancer types. Previously, miR-153 has been identified to be overexpressed in prostate cancer. miR-153 played an important role in promoting proliferation of human prostate cancer cells and presented a novel mechanism of microRNA-mediated direct suppression of phosphatase and tensin homolog (PTEN) expression in prostate cancer cells. Until now, little is known about the clinical significance of miR-153 expression in prostate cancer.The miR-153 expression in 143 pairs of prostate cancer and adjacent non-cancerous prostate tissues was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis. Student t test was conducted for intergroup comparison. Pearson correlation test was used for correlation analysis. Survival curves were carried out by the Kaplan-Meier method and evaluated using the log-rank test. Multivariable Cox proportional hazard risk regression model was performed to screen the independent factor affected the prognosis of prostate cancer patients.qRT-PCR analysis showed that the expression of miR-153 was significantly increased in the prostate cancer tissues in comparison with the adjacent noncancerous prostate tissues (P < .001). The high expression of miR-153 in prostate cancer tissues is closely correlated with aggressive clinical pathological parameters such as lymph node metastasis (P = .001); bone metastasis (P < .001); Gleason score (P < .001); and tumor-node-metastasis (TNM) stage (P < .001). Prostate cancer patients with a high expression of miR-153 had an evidently lower 5-year overall survival as compared with those with a low expression of miR-153 (P = .019). Notably, the multivariate Cox regression analysis indicated that miR-153 expression was an independent factor for predicting the 5-year overall survival of prostate cancer patients (hazard ratio [HR] = 2.481, 95% confidence interval [CI]: 1.582-10.727; P = .018).Our study demonstrated that high miR-153 expression was significantly associated with a poor overall survival independently of other factors in prostate cancer. Therefore, miR-153 may be an available biomarker for prostate cancer prognosis.


Assuntos
MicroRNAs/biossíntese , Neoplasias da Próstata/mortalidade , Neoplasias da Próstata/patologia , Idoso , Biomarcadores Tumorais , Intervalo Livre de Doença , Humanos , Estimativa de Kaplan-Meier , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Metástase Neoplásica , Prognóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa
4.
Biol Res ; 52(1): 53, 2019 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-31542051

RESUMO

BACKGROUND: Oxidative stress is the hallmark of diabetic encephalopathy, which may be caused by hyperglycaemic toxicity. We aimed to discover pharmacologic targets to restore redox homeostasis. We identified the transcription factor Nrf2 as such a target. METHODS: HT22 cells were cultured in 25 or 50 mM D-glucose with various concentrations of sulforaphane (SFN) (from 1.25 to 5.0 µM). Cell viability was tested with the Cell Counting Kit-8 assay. Reactive oxygen species (ROS) production was detected with an inverted fluorescence microscope using the dichlorodihydrofluorescein-diacetate fluorescent probe. The expression of NF-E2-related factor 2 (Nrf2), haem oxygenase-1 (HO-1) and nuclear factor-κB (NF-κB) at the mRNA and protein levels was detected by reverse transcription quantitative polymerase chain reaction and western blotting. RESULT: We found that a high glucose concentration (50 mM) increased the generation of ROS, downregulated the expression of Nrf2/HO-1 and upregulated the expression of NF-κB. Moreover, HT22 cell viability significantly decreased after culture in high-glucose medium for 24, 48 and 72 h, whereas the activation of the Nrf2/HO-1 pathway using a pharmacological Nrf2 activator abrogated this high-glucose-induced toxicity. CONCLUSION: This study suggests that the activation of the Nrf2-ARE signalling pathway might be a therapeutic target for the treatment of diabetic encephalopathy.


Assuntos
Glucose/toxicidade , Hipocampo/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/agonistas , Neuroproteção , Animais , Western Blotting , Linhagem Celular , Eletroforese em Gel de Campo Pulsado , Imunofluorescência , Hipocampo/citologia , Camundongos , Espécies Reativas de Oxigênio , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo
5.
Mem Inst Oswaldo Cruz ; 114: e190074, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31460570

RESUMO

BACKGROUND: Hepatitis delta virus (HDV) infections in hepatitis B virus (HBV) carriers are the most severe form of viral hepatitis. HDV prevalence is high in the Brazilian Amazon, but studies in other regions of the country are still scarce and often underestimated its prevalence by including a small numbers of individuals. OBJECTIVE: This study aimed to determine the serological prevalence of hepatitis D, the genotypes circulating and to evaluate the associated risk factors for acquisition of HDV in Minas Gerais state, Brazil. METHODS: We screened plasma samples (n = 498) from HBV chronic carriers for anti-HD antibodies using a commercial enzyme-linked immunosorbent assay (ELISA) kit. For those samples that were positive for anti-HD antibodies, we performed a reverse transcriptase (RT) nested-polymerase chain reaction (nested-PCR) in order to detect the viral genome and identify the viral genotypes circulating in the state. FINDINGS: The prevalence was 6.22% (31/498). Blood transfusion was the only risk factor associated with HDV infection [risk ratio: 3.73; 95% confidence interval (CI): 1.44 to 9.65]. For 26 anti-HD positive patients, HDAg gene sequences were determined and in all patients HDV genotype 1 was found. CONCLUSIONS: This study confirmed the circulation of HDV in Minas Gerais, an area previously considered non-endemic for hepatitis D in Brazil. The prevalence found in this study is much higher when compared to other studies performed in Brazil, probably because the population in our study was selected with minimal bias. Furthermore, in 26 anti-HD positive plasma samples, we were also able to detect the viral genome, indicating that these patients were experienced an active infection at the time of sample collection. These findings emphasise the importance of anti-HD testing in HBV infected individuals, which may contribute to this disease control in Brazil.


Assuntos
Anticorpos Anti-Hepatite/sangue , Hepatite B Crônica/epidemiologia , Hepatite D/epidemiologia , Vírus Delta da Hepatite , RNA Viral/genética , Adolescente , Adulto , Brasil/epidemiologia , Criança , Pré-Escolar , Estudos Transversais , Ensaio de Imunoadsorção Enzimática , Feminino , Genótipo , Hepatite B Crônica/complicações , Hepatite D/complicações , Hepatite D/diagnóstico , Vírus Delta da Hepatite/genética , Vírus Delta da Hepatite/imunologia , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Filogenia , Prevalência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Risco , Adulto Jovem
6.
J Appl Oral Sci ; 27: e20180256, 2019 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-31365706

RESUMO

OBJECTIVE: The rDNA-based method is unable to distinguish between alive and dead cells. Alternatively, bacterial viability can be assessed by molecular methods based on ribosomal RNA (rRNA). Therefore, this study aimed to detect viable streptococci in root canal samples using rRNA-based reverse transcription polymerase chain reaction (RT-PCR), compared to an rDNA-based PCR assay. METHODOLOGY: Microbiological root canal samples were obtained from 32 teeth with primary endodontic infections before (S1) and after chemomechanical preparation (S2), and after removal of intracanal medication (S3). RNA and DNA were extracted, and complementary DNA (cDNA) was synthesized from RNA using RT reaction. cDNA and genomic DNA were subjected to PCR with primers complementary to the 16S rRNA sequences of Streptococcus spp. McNemar's test was used to compare the detection rate of both assays (P<0.05). RESULTS: Streptococci were detected in 28.12% (9/32) and 37.5% (12/32) of S1 samples using rRNA- and rDNA-based PCR assays, respectively. In contrast, they were detected in only 6.25% (2/32) of S2 samples using rRNA-based RT-PCR, compared to 15.62% (5/32) using rDNA-based PCR. Finally, in S3 samples, streptococci were not detected by rRNA, whereas rDNA-based PCR still detected the bacteria in 12.5% (4/32) of the samples. The total number of PCR-positive reactions in the rDNA-based PCR was higher than in the rRNA-based assay (P<0.05). CONCLUSIONS: The rRNA-based RT-PCR showed a lower detection rate of streptococci when compared to the rDNA-based PCR, suggesting that the latter may have detected dead cells of streptococci in root canal samples.


Assuntos
DNA Ribossômico/isolamento & purificação , Cavidade Pulpar/microbiologia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Streptococcus/isolamento & purificação , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , DNA Ribossômico/genética , Humanos , RNA Bacteriano/genética , RNA Bacteriano/isolamento & purificação , RNA Ribossômico/genética , Reprodutibilidade dos Testes , Tratamento do Canal Radicular/métodos , Streptococcus/genética
7.
Medicine (Baltimore) ; 98(33): e16744, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31415369

RESUMO

MicroRNAs (miRNAs) play an important role in the pathogenesis of sepsis, but the association of miRNAs single nucleotide polymorphisms (SNPs) and sepsis risk is not clear. We analyzed plasma levels of miR-187, miR-21, and miR-145 in 180 patients with sepsis and 180 healthy controls were analyzed, and the SNPs: rs12605436, rs13137, and rs353291 were detected by sequencing. Plasma levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 were measured in all subjects by enzyme-linked immunosorbent assay (ELISA). The results showed that the levels of TNF-α and IL-6 in the plasma of patients with sepsis were significantly higher than those in patients of the control group (P < .0001). Plasma levels of miR-187 in patients with sepsis were significantly lower than those in the control group, while those of miR-21 and miR-145 were significantly higher than those in the control group (P < .0001). Plasma levels of miR-187 in sepsis patients were inversely correlated with those of TNF-α and IL-6 (r = -0.2841, -0.2163), and plasma levels of miR-21 and miR-145 were positively correlated with those of TNF-α and IL-6 (r = 0.615, 0.3057, 0.4465, 0.2734). The T allele of the miR-187 SNP rs12605436 was found to be a risk factor for sepsis (OR = 1.403, 95% CI = 1.205-1.612, P < .001). The T allele of the miR-21 SNP rs13137 and the T allele of the miR-145 SNP rs353291 (OR = 0.685, 95% CI = 0.566-0.820, P < .001) were found to be a protective factor for sepsis (OR = 0.755, 95% CI = 0.632-0.896, P < .001). From our results, we can see that the plasma levels of miRNAs containing the SNPs rs12605436, rs13137, and rs353291 are associated with the occurrence of sepsis.


Assuntos
Predisposição Genética para Doença , MicroRNAs/genética , Polimorfismo de Nucleotídeo Único , Sepse/genética , APACHE , Grupo com Ancestrais do Continente Asiático , Estudos de Casos e Controles , China , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interleucina-6/sangue , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Risco , Sepse/sangue , Fator de Necrose Tumoral alfa/sangue
8.
Pan Afr Med J ; 32: 202, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31312314

RESUMO

Introduction: Rotavirus causes severe-diarrheal diseases in infants. An estimation of 138 million rotavirus-associated diarrheal cases and 215,000 deaths occur every year globally. In December 2016, West-Shewa zone in Ethiopia reported unidentified gastrointestinal diarrhea outbreak. We investigated to identify the causative agent of the outbreak to support response operations. Methods: Medical records were reviewed, and the daily line list was collected from health facilities. Descriptive data analysis was done by time, person and place. Stool specimens were first tested by antigen capture enzyme immunoassay (EIA) technique and further confirmed by reverse-transcription polymerase chain reaction (RT-PCR) as a gold standard. The product of RT-PCR was genotyped for each gene using G1-G4, G8-G9 and G12 primers for VP7 gene and P(4), P(6), P(8) and P(14) primers for VP4 gene. Results: A total of 1,987 diarrheal cases (5.7 per 1000) and five deaths (case-fatality rate 0.25%) were identified and epidemiologically-linked to confirmed rotavirus from December 2016 to February 2017. Among the cases, 1,946 (98%) were < 5 children. Fourteen (74%) of the 19 tested stool specimens were positive for rotavirus by EIA and RT-PCR. Majority of strains detected were G12P(6) (25%) and G-negative P(8) (25%) followed by G9P(8) (19%), G1P(8) (13%) and G3/G2 P(8), G12P(8), and G-negative P(6) (6% each). Conclusion: Diarrheal outbreak which occurred in West-Shewa zone of Ethiopia was associated with rotavirus and relatively more affected districts with low vaccination coverage. Routine rotavirus vaccination quality and coverage should be evaluated and the surveillance system needs to be strengthened to detect, prevent and control a similar outbreak.


Assuntos
Diarreia/epidemiologia , Infecções por Rotavirus/epidemiologia , Vacinas contra Rotavirus/administração & dosagem , Rotavirus/isolamento & purificação , Doença Aguda , Adolescente , Adulto , Criança , Pré-Escolar , Diarreia/virologia , Surtos de Doenças , Etiópia/epidemiologia , Fezes/virologia , Feminino , Genótipo , Humanos , Técnicas Imunoenzimáticas/métodos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rotavirus/genética , Infecções por Rotavirus/virologia , Cobertura Vacinal/estatística & dados numéricos , Adulto Jovem
9.
Arch Virol ; 164(10): 2537-2543, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31309291

RESUMO

In May 2017, many free-ranging dorcas gazelles (Gazella dorcas) with suspected signs of peste des petits ruminants (PPR) were reported in Dinder National Park, South-Eastern Sudan. Peste des petits ruminants virus (PPRV) antigen and nucleic acid were detected in specimens from these gazelles using an immunocapture ELISA and a reverse transcription polymerase chain reaction (RT-PCR) assays. PPRV was also detected in four healthy semi-captive dorcas gazelles from two areas of Khartoum State. Phylogenetic analysis showed that these PPRV strains belonged to the lineage IV genotype. The present study demonstrates that gazelles are a potential wild small ruminant host for PPRV and may influence the epidemiology of PPR in the Sudan.


Assuntos
Antílopes/virologia , Reservatórios de Doenças , Peste dos Pequenos Ruminantes/virologia , Vírus da Peste dos Pequenos Ruminantes/isolamento & purificação , Animais , Antígenos Virais/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Genótipo , Vírus da Peste dos Pequenos Ruminantes/classificação , Vírus da Peste dos Pequenos Ruminantes/genética , Filogenia , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Gastropatias , Sudão
10.
Arch Virol ; 164(10): 2493-2504, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31346769

RESUMO

One of the pathological forms of chronic hepatitis C is occult HCV infection (OCI), in which there is no detectable HCV RNA in plasma specimens but HCV RNA is present in PBMCs and liver biopsy specimens. The aim of this study is to estimate the prevalence of OCI in HIV-positive people who are injection drug users (IDUs). From April 2015 to August 2018, 161 Iranian IDUs with HIV infection enrolled in the study. Viral RNA was extracted from plasma and PBMC samples of participants, and the presence of HCV RNA was examined using RT nested PCR with primers from two conserved regions (5´-UTR and NS5B). HCV genotyping was performed using RFLP and sequencing methods. Of the 161 patients, 134 (83.2%) were positive for anti-HCV antibodies. All 27 patients who were negative for anti-HCV were also negative for HCV RNA in plasma, but five of them (18.5%) were positive for HCV RNA in PBMCs. Importantly, 9 out of 50 patients (18.0%) who apparently had recovered from HCV infection (i.e., were anti-HCV positive and HCV RNA negative) were positive for HCV RNA in PBMCs. Overall, 18.1% of the patients who had no signs of previous HCV infection or had apparently recovered from the disease had OCI. The HCV genotypes of the cases with OCI were as follows: five patients (35.7%) were infected with subtype 1a, eight patients (57.1%) were infected with subtype 3a, and one patient (7.1%) was infected with genotype 4. Thus, it seems that the prevalence of OCI in HIV-positive IDUs is extremely significant in Iran and is likely to delay the global eradication of HCV infection until 2030.


Assuntos
Usuários de Drogas , Infecções por HIV/complicações , Hepatite C Crônica/epidemiologia , Hepatite C Crônica/patologia , RNA Viral/sangue , Abuso de Substâncias por Via Intravenosa/complicações , Adulto , Idoso , Feminino , Genótipo , Técnicas de Genotipagem , Hospitais Universitários , Humanos , Irã (Geográfico) , Leucócitos Mononucleares/virologia , Masculino , Pessoa de Meia-Idade , Plasma/virologia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Prevalência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto Jovem
11.
Arch Virol ; 164(10): 2573-2580, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31346770

RESUMO

Recently, five new viruses from the genus Vitivirus were identified and named grapevine virus G, H, I, J and L. These viruses were targeted in a survey to evaluate their prevalence in different grapevine populations in California. Excluding a single detection of GVJ, other vitiviruses were detected infecting several grapevine selections via RT-PCR and later confirmed by sequencing. This paper represents the first report of GVG, GVH and GVI in California. In a preliminary analysis, the sequence diversity between identified isolates of GVG, GVH, GVI and GVL was investigated using distance matrices and phylogenetics. Finally, coinfections involving diverse vitiviruses and leafroll viruses were evidenced.


Assuntos
Flexiviridae/classificação , Flexiviridae/isolamento & purificação , Variação Genética , Doenças das Plantas/virologia , Vitis/virologia , California , Flexiviridae/genética , Prevalência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA
12.
World J Microbiol Biotechnol ; 35(7): 101, 2019 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-31236717

RESUMO

Propionibacterium acidipropionici produces propionic acid from different sugars and glycerol; the production can be improved by high cell density fermentations using immobilized cells that help to overcome the limitations of the non-productive lag phase and product inhibition. In this study, the use of stress factors to induce P. acidipropionici to form biofilm and its use as an immobilization procedure in fermentations in bioreactors for producing propionic acid was investigated. Citric acid and sodium chloride increased exopolysaccharide production, biofilm forming capacity index and trehalose production. Analysis of the expression of trehalose synthesis-related genes otsA and treY by RT-qPCR showed significantly increased expression of only treY during log phase with citric acid, while FISH analysis showed expression of treY and luxS under the influence of both stress factors. The stress factors were then used for development of microbial biofilms as immobilization procedure on Poraver® and AnoxKaldnes® carriers in recycle batch reactors for propionic acid production from 20 g/L glycerol. Highest productivities of 0.7 and 0.78 g/L/h were obtained in Poraver® reactors, and 0.39 and 0.43 g/L/h in AnoxKaldnes® reactors with citric acid and NaCl, respectively.


Assuntos
Biofilmes , Propionatos/metabolismo , Propionibacterium/metabolismo , Estresse Fisiológico , Reatores Biológicos/microbiologia , Células Imobilizadas , Fermentação , Regulação da Expressão Gênica , Genes Bacterianos , Glicerol/metabolismo , Hibridização in Situ Fluorescente , Microscopia de Força Atômica , Propionibacterium/genética , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trealose/metabolismo
13.
Braz J Med Biol Res ; 52(6): e8589, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31166385

RESUMO

The transport of myo-inositol is the main mechanism for the maintenance of its high intracellular levels. We aimed to measure the mRNA and protein levels of myo-inositol cotransporters in the sciatic nerve (SN) and dorsal root ganglia (DRG) during experimental diabetes. Streptozotocin-induced (STZ; 4, 8, and 12 weeks; 65 mg/kg; ip) diabetic rats (DB) and age-matched euglycemic (E) rats were used for the analysis of mRNA and protein levels of sodium myo-inositol cotransporters 1, 2 (SMIT1, SMIT2) or H+/myo-inositol cotransporter (HMIT). There was a significant reduction in the mRNA levels for SMIT1 in the SN and DRG (by 36.9 and 31.0%) in the 4-week DB (DB4) group compared to the E group. SMIT2 was not expressed in SN. The mRNA level for SMIT2 was up-regulated only in the DRG in the DB4 group. On the other hand, the protein level of SMIT1 decreased by 42.5, 41.3, and 44.8% in the SN after 4, 8, and 12 weeks of diabetes, respectively. In addition, there was a decrease of 64.3 and 58.0% of HMIT in membrane and cytosolic fractions, respectively, in the SN of the DB4 group. In the DRG, there was an increase of 230 and 86.3% for SMIT1 and HMIT, respectively, in the DB12 group. The levels of the main inositol transporters, SMIT1 and HMIT, were greatly reduced in the SN but not in the DRG. SMIT-1 was selectively reduced in the sciatic nerve during experimental STZ-induced diabetes.


Assuntos
Transporte Biológico Ativo/fisiologia , Diabetes Mellitus Experimental/metabolismo , Gânglios Espinais/metabolismo , Inositol/metabolismo , RNA Mensageiro/metabolismo , Nervo Isquiático/metabolismo , Animais , Western Blotting , Masculino , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estreptozocina , Regulação para Cima
14.
Rev Soc Bras Med Trop ; 52: e20190060, 2019 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-31188919

RESUMO

INTRODUCTION: In this study, we aimed to identify DENV-2 subtypes in Aedes aegypti pools collected between 2011 and 2017 in a rural area of Northern Cordoba, Colombia ("La Balsa"). METHODS: RT-PCR was performed to analyze the capsid/pre-membrane region (C-PrM). Sequencing and phylogenetic bayesian inference using reference DENV-2 sequences were performed. RESULTS: Twelve pools that tested positive for DENV-2 were characterized based on the C-PrM region and grouped under the Asian/American clade. CONCLUSIONS: This study is the first to report the DENV-2 Asian-American subtype in a rural area of Cordoba region, which is associated with severe dengue and local epidemics.


Assuntos
Aedes/virologia , Vírus da Dengue/classificação , Vírus da Dengue/genética , Dengue/virologia , Mosquitos Vetores/virologia , Filogenia , Animais , Teorema de Bayes , Colômbia/epidemiologia , Dengue/epidemiologia , Vírus da Dengue/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sorogrupo , Sorotipagem , Dengue Grave
15.
Acta Vet Scand ; 61(1): 31, 2019 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-31234899

RESUMO

BACKGROUND: The small, single-stranded positive-sense RNA astroviruses are mostly known to be enteric viruses. In recent years, though, different astroviruses were reported in association with neurological disease in various species. In cattle, two distinct neurotropic astrovirus genotype species were described in numerous cases of nonsuppurative encephalomyelitis, with one of these viruses also reported in similar circumstances in several sheep. Here, we retrieved archived formalin-fixed, paraffin-embedded brain tissues of a muskox diagnosed with a comparable disease pattern in 1982 and investigated them for the presence of neurotropic astroviruses with various techniques. RESULTS: Initially, tissue samples scored positive for both neurotropic astroviruses by immunohistochemistry; however, unexpected results with further immunohistochemical testing, in situ hybridization and qRT-PCR prompted us to submit an RNA extract from the animal's brain material to next-generation sequencing. We were thus able to obtain the full genome of a novel astrovirus, muskox astrovirus CH18 (MOxAstV-CH18), whose closest relative is an enteric ovine astrovirus. Subsequently, viral RNA could be detected with a specific RT-PCR in the brain of the affected animal, but not in faecal samples from the current muskoxen herd of the animal park where the animal used to be kept. CONCLUSIONS: We identified a novel astrovirus in a historical case of a captive muskox with nonsuppurative encephalomyelitis. Unfortunately, our results and the fact that no material from organs other than of the nervous system was available do not allow any assumption about the epidemiology or pathogenesis of the virus. Still, these findings are yet another piece of evidence that the tropism and species specificity of astroviruses could be more deceptive than generally assumed.


Assuntos
Infecções por Astroviridae/veterinária , Astroviridae/fisiologia , Ruminantes/virologia , Animais , Astroviridae/classificação , Infecções por Astroviridae/virologia , Genoma Viral/genética , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa
16.
J Med Microbiol ; 68(8): 1240-1243, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31237533

RESUMO

The aim of the present study was to report the molecular characterization of human group A rotaviruses (RVAs) circulating in Tunisia. Stool specimens were collected from children under 5 years of age who had been hospitalized or were consulting for gastroenteritis in Tunisian hospitals between 2015 and 2017. All samples were screened by reverse-transcription polymerase chain reaction (RT-PCR) for the detection of the VP6 gene specific for RVA. RVA-positive samples were further analysed for G/P genotyping by semi-nested multiplex RT-PCR. Among 454 tested samples, 72 (15.8 %) were positive for RVA. G1P[8] was the most prevalent detected strain (41.7%), followed by G9P[8] (32.8%), G2P[4] (7.5%), G12P[8] (7.5%), G1P[6] (3.0%), G2P[8] (1.5%) and G3P[8] (1.5%), with mixed infections in 4.5 % of cases. In the absence of a national anti-rotavirus vaccination strategy, RVAs remain the primary aetiological agent for gastroenteritis in Tunisian children.


Assuntos
Gastroenterite/virologia , Infecções por Rotavirus/virologia , Rotavirus/genética , Proteínas do Capsídeo/genética , Pré-Escolar , Fezes/virologia , Gastroenterite/epidemiologia , Variação Genética , Genótipo , Humanos , Lactente , Recém-Nascido , Epidemiologia Molecular , Prevalência , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologia , Estações do Ano , Análise de Sequência de DNA , Tunísia/epidemiologia
17.
J Appl Oral Sci ; 27: e20180641, 2019 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-31166414

RESUMO

OBJECTIVES: Infection, inflammation and bone resorption are closely related events in apical periodontitis development. Therefore, we sought to investigate the role of cyclooxygenase (COX) in osteoclastogenesis and bone metabolism signaling in periapical bone tissue after bacterial lipopolysaccharide (LPS) inoculation into root canals. METHODOLOGY: Seventy two C57BL/6 mice had the root canals of the first molars inoculated with a solution containing LPS from E. coli (1.0 mg/mL) and received selective (celecoxib) or non-selective (indomethacin) COX-2 inhibitor. After 7, 14, 21 and 28 days the animals were euthanized and the tissues removed for total RNA extraction. Evaluation of gene expression was performed by qRT-PCR. Statistical analysis was performed using analysis of variance (ANOVA) followed by post-tests (α=0.05). RESULTS: LPS induced expression of mRNA for COX-2 (Ptgs2) and PGE2 receptors (Ptger1, Ptger3 and Ptger4), indicating that cyclooxygenase is involved in periapical response to LPS. A signaling that favours bone resorption was observed because Tnfsf11 (RANKL), Vegfa, Ctsk, Mmp9, Cd36, Icam, Vcam1, Nfkb1 and Sox9 were upregulated in response to LPS. Indomethacin and celecoxib differentially modulated expression of osteoclastogenic and other bone metabolism genes: celecoxib downregulated Igf1r, Ctsk, Mmp9, Cd36, Icam1, Nfkb1, Smad3, Sox9, Csf3, Vcam1 and Itga3 whereas indomethacin inhibited Tgfbr1, Igf1r, Ctsk, Mmp9, Sox9, Cd36 and Icam1. CONCLUSIONS: We demonstrated that gene expression for COX-2 and PGE2 receptors was upregulated after LPS inoculation into the root canals. Additionally, early administration of indomethacin and celecoxib (NSAIDs) inhibited osteoclastogenic signaling. The relevance of the cyclooxygenase pathway in apical periodontitis was shown by a wide modulation in the expression of genes involved in both bone catabolism and anabolism.


Assuntos
Inibidores de Ciclo-Oxigenase/farmacologia , Cavidade Pulpar/metabolismo , Lipopolissacarídeos/farmacologia , Osteogênese/fisiologia , Tecido Periapical/efeitos dos fármacos , Tecido Periapical/metabolismo , Prostaglandina-Endoperóxido Sintases/fisiologia , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Reabsorção Óssea/metabolismo , Celecoxib/farmacologia , Ciclo-Oxigenase 2/análise , Escherichia coli/metabolismo , Expressão Gênica , Indometacina/farmacologia , Lipopolissacarídeos/análise , Masculino , Camundongos Endogâmicos C57BL , Osteogênese/efeitos dos fármacos , Prostaglandina-Endoperóxido Sintases/análise , Prostaglandina-Endoperóxido Sintases/efeitos dos fármacos , Receptores de Prostaglandina E/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Regulação para Cima
18.
Plant Sci ; 285: 1-13, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31203874

RESUMO

Bioactive gibberellins (GAs) play multiple roles in plant development and stress responses. GA2-oxidases (GA2oxs) are a class of 2-oxoglutarate-dependent dioxygenases that regulate the deactivation of bioactive GAs. In this study, we investigated the phylogeny and domain structures of the seven GA2ox genes present in the Arabidopsis thaliana genome. Comprehensive expression analysis using translational reporter lines showed that the seven GA2ox genes are differentially expressed during Arabidopsis growth and development: GA2ox1 is specifically expressed in the hypocotyl and lateral root primordium; GA2ox2 is highly expressed in aboveground tissues; GA2ox3 is expressed in the chalazal endosperm of the early embryo sac and inflorescences; GA2ox4 is expressed in the shoot apical meristem and during lateral root initiation; GA2ox6 is expressed in the maturation zone, but not in the meristem or elongating zone of the root; GA2ox7 is constitutively expressed during almost all developmental stages; and GA2ox8 is exclusively expressed in stomatal cells. Overexpression of each of these GA2ox genes inhibited high temperature-induced hypocotyl elongation in both wild-type and elongated hypocotyl 5 plants, which have an elongated hypocotyl phenotype, suggesting that these genes negatively regulate hypocotyl elongation by reducing bioactive GA levels. This study provides a valuable resource for further elucidating the roles of GA2ox genes during different stages of development.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Genes de Plantas/fisiologia , Giberelinas/metabolismo , Oxirredutases/genética , Arabidopsis/enzimologia , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Giberelinas/fisiologia , Hipocótilo/crescimento & desenvolvimento , Hipocótilo/metabolismo , Oxirredutases/metabolismo , Oxirredutases/fisiologia , Filogenia , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Transcriptoma
19.
Plant Sci ; 285: 248-257, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31203890

RESUMO

Halophytic Oryza coarctata is a good model system to examine mechanisms of salinity tolerance in rice. O. coarctata leaves show the presence of microhairs in adaxial leaf surface furrows that secrete salt under salinity. However, detailed molecular and physiological studies of O. coarctata microhairs are limited due to their relative inaccessibility. This work presents a detailed characterization of O. coarctata leaf features. O. coarctata has two types of microhairs on the adaxial leaf surface: longer microhairs (three morphotypes) lining epidermal furrow walls and shorter microhairs (reported first time) arising from bulliform cells. Microhair morphotypes include (i) finger-like, tubular structures, (ii) tubular hairs with bilobed and flattened heads and (iii) bi-or trifurcated hairs. The unicellular nature of microhairs was confirmed by propidium iodide (PI) staining. An efficient method for the isolation and enrichment of O. coarctata microhairs is presented (yield averaging ˜2 × 105/g leaf tissue). The robustness of the microhair isolation procedure was confirmed by subsequent viability staining (PI), total RNA isolation and RT-PCR amplification of O. coarctata trichome-specific WUSCHEL-related homeobox 3B (OcWox3B) and transporter gene-specific cDNA sequences. The present microhair isolation work from O. coarctata paves the way for examining genes involved in ion secretion in this halophytic wild rice model.


Assuntos
Oryza/anatomia & histologia , Folhas de Planta/anatomia & histologia , Plantas Tolerantes a Sal/anatomia & histologia , Microscopia Confocal , Oryza/fisiologia , Folhas de Planta/citologia , Folhas de Planta/fisiologia , Folhas de Planta/ultraestrutura , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Plantas Tolerantes a Sal/fisiologia , Tricomas/anatomia & histologia , Tricomas/fisiologia , Tricomas/ultraestrutura
20.
Braz Oral Res ; 33: e035, 2019 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-31038569

RESUMO

Eugenia dysenterica is a Brazilian tree investigated for its properties and bioactive compounds, which are believed to have both pharmacological and phytochemical therapeutic effects. The leaves of this tree contain tannins, flavonoids, terpenes, and saponins, with reportedly beneficial effects to the human body. Despite these therapeutic applications, its effects have never been tested on oral tissues. Therefore, the aim of the present study was to evaluate the cytotoxic and antioxidant effects and the anti-inflammatory and repair properties of the acetone fraction of E. dysenterica on primary culture of human gingival fibroblasts and on the immortalized murine macrophage cell line (RAW 264.7). For this purpose, a metabolic activity assay, a wound healing assay, a nitric oxide assay, and RT-qPCR were performed. The assays revealed a cytoprotective effect of this plant, suggested by the increase in the expression of SOD1 and NRF2. An antioxidant potential effect was observed in the DPPH• assay. However, the fraction of E. dysenterica did not show anti-inflammatory activity. In conclusion, Eugenia dysenterica may promote cytoprotection when associated with chlorhexidine digluconate because of its antioxidant effect. However, additional studies are necessary on other human dental tissues using other parts of the plant in order to develop a possible mouthwash to assist patients with oral disorders.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Eugenia/química , Fibroblastos/efeitos dos fármacos , Gengiva/citologia , Extratos Vegetais/farmacologia , Animais , Brasil , Células Cultivadas , Clorexidina/análogos & derivados , Clorexidina/farmacologia , Humanos , Camundongos , Óxido Nítrico/análise , Folhas de Planta/química , Células RAW 264.7 , Valores de Referência , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Cicatrização/efeitos dos fármacos
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