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1.
J Chromatogr A ; 1700: 464049, 2023 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-37178554

RESUMO

The aim of this work is to investigate the efficiency of a phosphonium-based strong anion exchange sorbent for the extraction of some selected phenolic acids. The material was synthesized through chloromethylation of a porous poly(styrene-divinylbenzene) substrate with high degree of crosslinking, followed by quaternarization with tributyl phosphine. The parameters affecting the solid phase extraction of five phenolic acids, namely chlorogenic acid, caffeic acid, dihydroxybenzoic acid, ferulic acid and rosmarinic acid were optimized. The sample pH and the type, volume and concentration of the eluting solutions were investigated. The analysis of the phenolic acids after extraction was performed using HPLC with diode array detection. Limit of detection, limit of quantitation, linear range, correlation coefficient and reproducibility for the determination of the phenolic acids were estimated. The retention of the phenolic acids on the developed phase was studied using breakthrough analysis. The experimental breakthrough curves were fitted by Boltzmann's function, and the regression parameters were utilized for the determination of the breakthrough parameters. The results obtained using the developed phase were compared with those obtained by the commercially available Oasis MAX sorbent. The proposed approach was successfully applied for the extraction and pre-concentration of rosmarinic acid from rosemary leaf (Rosmarini folium) alcoholic extract.


Assuntos
Resinas de Troca Aniônica , Extração em Fase Sólida , Reprodutibilidade dos Testes , Extração em Fase Sólida/métodos , Cromatografia Líquida de Alta Pressão/métodos , Ácido Rosmarínico
2.
Egypt J Immunol ; 29(4): 163-173, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36208045

RESUMO

The outcome for chronic phase (CP) chronic myelogenous leukemia (CML) patients has changed dramatically since the introduction of tyrosine kinase inhibitor (TKI) therapy. We examined the characteristics of CML patients during TKI therapy by determining the plasma concentrations of soluble vascular cell adhesion molecule 1 (sVCAM-1), and transforming growth factor (TGFß1) biomarkers. The plasma levels of sVCAM-1 and TGFß1 were measured by ELISA at baseline and after 3 months of TKI treatment. The levels of sVCAM-1, and TGFß1 were significantly elevated in patients with CML (P< 0.01). Dasatinib treatment was associated with a significant reduction in the levels of these biomarkers (P< 0.01). In conclusion, plasma levels of sVCAM-1 and TGFß1 could have a role in the pathogenesis of CML and may be used as predictors of hematological and molecular responses to TKIs. A favorable outcome for Dasatinib therapy was observed.


Assuntos
Leucemia Mielogênica Crônica BCR-ABL Positiva , Fator de Crescimento Transformador beta1/sangue , Molécula 1 de Adesão de Célula Vascular , Biomarcadores , Dasatinibe/farmacologia , Dasatinibe/uso terapêutico , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/induzido quimicamente , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico
3.
Discov Oncol ; 13(1): 86, 2022 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-36083385

RESUMO

BACKGROUND: FOXP3 and ROR-γ genes are master regulators of the Treg and Th17 differentiation, respectively. This work was planned to investigate the impact of FOXP3 (rs3761548C/A and rs3761549C/T) and ROR-γ (rs9017A/G & rs9826A/G) gene polymorphism on the vulnerability of pediatric Egyptians to acute lymphoblastic leukemia (ALL). Furthermore, we evaluated the impact of these genetic variations on Treg/Th17-related cytokines. METHODS: FOXP3 SNPs were genotyped using PCR-based restriction fragment length polymorphism (PCR-RFLP), while ROR-γ SNPs polymorphism were performed by PCR-sequence-specific primer (PCR-SSP). An Enzyme-linked immunosorbent assay (ELISA) was used to assess the levels of Treg/Th17 associated cytokines on 128 ALL children and 124 healthy donors. RESULTS: Compared to controls, patients had a significant increase (p < 0.01/p < 0.05) in FOXP3rs3761548CC genotype and a significant decrease (p < 0.001/p < 0.01) inrs3761548CA genotype. A significant elevation (p < 0.001/p < 0.01) in ROR-γ rs9017AA genotype and a significant reduction (p < 0.01/p < 0.05) in rs9017AG genotype were detected in ALL patients versus controls. An insignificant change in FOXP3 (rs3761549C/T) and ROR-γ (rs9826A/G) genotypes was demonstrated between both groups. ROR-γ GG and GA haplotypes were significantly decreased (p < 0.05/p < 0.05; p < 0.05/p < 0.05) in ALL subjects compared to healthy ones. Relapsed patients had a significantly higher (p < 0.05/P < 0.05) frequency of FOXP3 rs3761548CA genotype than non-relapsed subjects. ROR-γ rs9017AG and rs9826GG genotypes might be associated with the increase in IL-23 plasma level. CONCLUSIONS: Our preliminary data provided evidence for the impact ofFOXP3 (rs3761548C/A) and ROR-γ (rs9017A/G) gene polymorphisms and the occurrence of ALL in Egyptian children. Another large-scale prospective study should be conducted to validate these findings.

4.
Vaccines (Basel) ; 10(9)2022 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-36146540

RESUMO

Coronavirus disease 2019 (COVID-19) has affected millions of people worldwide. During the early stages of vaccination in Egypt, the ChAdOx1 nCoV-19 and BBIBP-CorV vaccines were the most distributed. The aim of this study was to compare the immune responses and short-term efficacies of these two vaccines. We recruited adults who received two doses of either vaccine. Samples were collected after the first dose of ChAdOx1 nCoV-1 and after the second dose of both vaccines. Antibodies against SARS-CoV-2 antigens were measured using LABScreen™ COVID Plus kits, and cell-mediated immune responses were assessed using flow cytometry. Of the 109 recruited subjects, 60 (55%) received the ChAdOx1 nCoV-19 vaccine, and the remainder received the BBIBP-CorV vaccine. The total antibody level did not significantly differ between the two groups. The level of the anti-spike subunit 2 (S2) antibody was significantly higher in the ChAdOx1 nCoV-19 group. The percentages of both total T cells and B cells were unaffected by the type of vaccination. However, the ChAdOx1 nCoV-1 vaccine was significantly associated with a higher percentage of CD8+ cells. The vaccines did not significantly differ in the number or severity of infections postvaccination. None of the participants were admitted to the hospital or died of COVID-19 infection. In conclusion, the BBIBP-CorV vaccine is associated with an immune response and protection against infection that is comparable to that of the ChAdOx1 nCoV-1 vaccine. Follow-up is needed to study the long-term protective effects of both vaccines. Inactivated vaccines are easier to manufacture in developing countries and their limited side effects may lead to better economic benefits by limiting the number of absences from work.

5.
Analyst ; 147(3): 534-541, 2022 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-35060566

RESUMO

Porous layered monolithic substrates of poly(butyl methacrylate-co-ethylene dimethacrylate) were synthesized via UV initiated free radical polymerization in the presence of fluoroponytailed ionic liquids as co-porogenic constituents. The effects of the type and the amount of selected fluorous ionic liquids on various properties of the monolithic support, e.g. porosity, specific surface area and chromatographic performance, in particular for their usability in reversed phase TLC, were examined. Porosity was characterized by means of mercury porosimetry and scanning electron microscopy. The monolithic stationary phases with different layer thickness were successfully applied in the separation of three curcuminoids, namely curcumin, demethoxycurcumin and bisdemethoxycurcumin. Relative retention factor, theoretical plates and resolution were used for the evaluation of the monolithic support's performance. To verify the feasibility of the monoliths, the method was employed for the discrimination between the plant species Curcuma longa and Curcuma xanthorrhiza.


Assuntos
Líquidos Iônicos , Cromatografia em Camada Fina , Metacrilatos , Polimerização , Porosidade
6.
Hum Immunol ; 83(1): 10-16, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34607724

RESUMO

Genetic differences among individuals could affect the clinical presentations and outcomes of COVID-19. Human Leukocyte Antigens are associated with COVID-19 susceptibility, severity, and prognosis. This study aimed to identify HLA-B and -C genotypes among 69 Egyptian patients with COVID-19 and correlate them with disease outcomes and other clinical and laboratory data. HLA-B and -C typing was performed using Luminex-based HLA typing kits. Forty patients (58%) had severe COVID-19; 55% of these patients died, without reported mortality in the moderate group. The alleles associated with severe COVID-19 were HLA-B*41, -B*42, -C*16, and -C*17, whereas HLA-B*15, -C*7, and -C*12 were significantly associated with protection against mortality. Regression analysis showed that HLA-B*15 was the only allele associated with predicted protection against mortality, where the likelihood of survival increased with HLA-B*15 (P < 0.001). Patient survival was less likely to occur with higher total leukocytic count, ferritin, and creatinine levels. This study provides interesting insights into the association between HLA class I alleles and protection from or severity of COVID-19 through immune response modulation. This is the first study to investigate this relationship in Egyptian patients. More studies are needed to understand how HLA class I alleles interact and affect Cytotoxic T lymphocytes and natural killer cell function.


Assuntos
COVID-19/genética , Antígeno HLA-B15/genética , SARS-CoV-2/patogenicidade , Idoso , COVID-19/imunologia , COVID-19/mortalidade , COVID-19/virologia , Egito , Feminino , Predisposição Genética para Doença , Antígeno HLA-B15/imunologia , Haplótipos , Interações Hospedeiro-Patógeno , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Fatores de Proteção , Medição de Risco , Fatores de Risco , SARS-CoV-2/imunologia , Índice de Gravidade de Doença , Fatores de Tempo
7.
Mol Immunol ; 124: 1-8, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32480291

RESUMO

Regulatory T cells (Tregs) is one of the immunosuppressive subsets of CD4+ T cells characterized by transcription factor forkhead box protein P3 (FOXP3) expression which are involved in tumor development and progression. Identification of the factors that influence Treg cell function is extremely important. Our current study aimed to evaluate the frequency of Treg cells, cytokine secretion and the expression of microRNAs (miRNAs) in pediatric acute lymphoblastic leukemia (ALL) patients. The frequency of CD3+, CD4+ and CD4+CD25+FOXP3+ Treg was assessed by flow cytometry in 43 ALL patients versus 42 controls. Plasma levels of IL-10, transcription factor ß (TGF-ß), IL-6, IL-17, IL-23 and tumor necrosis factor (TNF-α) were measured by Enzyme-linked immunosorbent assay (ELISA). miR-21, miR-24, miR-26a, miR133b, miR-148a and miR-155 expression were analyzed using quantitative real-time polymerase chain reaction (qRT-PCR). A slight insignificant increase in Treg cells in ALL patients compared to controls was observed. There was a significant elevation in IL-10 (p < 0.05), IL-6 (p < 0.01), IL-23 (p < 0.05) and TNF-α (p < 0.01) in ALL patients compared with controls. Meanwhile, a significant reduction in TGF-ß (p < 0.001) was recorded. A slight insignificant decrease in IL-17 in ALL patients was observed.ALL patients showed a significant increase in miR-21 (p < 0.05), miR-148a (p < 0.01), miR-24 (p < 0.05) and a significant reduction in miR-155 (p < 0.01). In conclusion, the slight change in Treg cells frequency and alteration in related cytokines could possibly involve in the pathogenesis of ALL. Dysregulated miRNAs, as a regulatory mechanism of epigenetics, might contribute to these observed results. Further researches are required to confirm our interesting findings.


Assuntos
Citocinas/imunologia , MicroRNAs/imunologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/imunologia , Linfócitos T Reguladores/imunologia , Criança , Pré-Escolar , Feminino , Fatores de Transcrição Forkhead/imunologia , Regulação da Expressão Gênica/imunologia , Humanos , Lactente , Subunidade alfa de Receptor de Interleucina-2/imunologia , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética
8.
Pathol Oncol Res ; 26(2): 1183-1190, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31187468

RESUMO

Cyclooxygenase-2 (COX-2) plays an important role in carcinogenesis, which catalyzes the conversion of arachidonic acid into prostaglandins. P53 is a tumor suppressor gene that contributes to apoptosis and cell cycle control. There is functional interaction between p53 and COX-2, which lead to abrogation of apoptosis and progression of malignancy. To assess the relationship between COX-2, p53 expression and the clinicopathololgic features in SLL and DLBCL. We immunohistochemically examined the expression of COX-2 and p53 in non-neoplastic lymphoid cells, lymph nodal low-grade (50 cases of SLL), intermediate and high-grade lymphomas (100 cases of DLBCL) and their corresponding bone marrow specimens. The expression of COX-2 and p53 was absent in the in non-neoplastic lymphoid cells. In contrast, their expression values increased progressively with the advancing grade of lymphoma (p < 0.001). COX-2 expression was significantly associated with advanced disease stage, high-grade lymphomas, and disease relapse and p53 expression. The p53was detected in 64.5% in patients positive for COX-2. The expressions of COX-2 and p53 proteins, were significantly associated with shorter overall-survival and progression free survival. Here we report up-regulation of COX-2and p53 protein expression in SLL and DLBCL indicating their interactive involvement in the pathogenesis of lymphoma. Our data provide a rationale for further investigation of COX-2 expression in lymphomas for potential prognostic, chemopreventive and chemotherapeutic purposes.


Assuntos
Ciclo-Oxigenase 2/biossíntese , Linfoma de Células B/patologia , Proteína Supressora de Tumor p53/biossíntese , Adulto , Idoso , Biomarcadores Tumorais/análise , Feminino , Humanos , Linfoma de Células B/mortalidade , Masculino , Pessoa de Meia-Idade , Prognóstico , Intervalo Livre de Progressão
9.
Indian J Hematol Blood Transfus ; 35(3): 471-477, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31388259

RESUMO

Non-Hodgkin's lymphoma (NHL) is an exceedingly diversified group of lymphoproliferative neoplasms emerging from B-, T- or natural killer -lymphocytes. This study was done to detect Matrix metalloproteinase-2 (MMP2)-735C/T gene polymorphism in patients with NHL and its relation to the clinicopathological characteristics of the studied patients in addition to detection the association between it and NHL disease susceptibility and progression. Clinico-hematological profiles were done on 50 NHL patients. The genotypes and allelic frequencies of MMP-2 polymorphisms were recognized utilizing Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP). PCR products after adding restriction endonuclease were analyzed using QIAxcel advanced (automated) instrument. The CT + TT genotypes and T allele of MMP2 735C/T were statistically significant in patients having advanced clinical stages III/IV compared to patients with stages I/II. Another significance was observed in patients with intermediate high/high IPI score and BM infiltration. Interestingly, patients with MMP2-735C/T genotype exhibit lower rate of survival. Our results demonstrated that MMP2-735C/T polymorphism may potentially affect the progression of NHL. Further larger scale studies are needed.

10.
Talanta ; 194: 542-547, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30609570

RESUMO

A simple offline coupling voltammetry-MALDI/TOF MS procedure is presented for studying electrochemical reactions. It was utilized for the characterization of the electro-reduction products of febuxostat in methanolic acetate buffer (0.1 M, pH 5). The MS analysis reveals that the carboxylic and nitrile groups are the electro-reducible groups at -0.9338 and -1.5503 V with the conversion to aldehydic and amino groups, respectively. The developed voltammetric method was validated and applied successfully for the drug determination in pharmaceutical tablets and real plasma samples within the linearity ranges 0.03-2 and 0.4-5 µg mL-1, respectively.


Assuntos
Análise Química do Sangue/métodos , Eletroquímica/métodos , Febuxostat/sangue , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Ácidos Carboxílicos/química , Febuxostat/química , Febuxostat/metabolismo , Humanos , Nitrilas/química , Xantina Oxidase/metabolismo
11.
J Pharm Biomed Anal ; 162: 117-123, 2019 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-30236819

RESUMO

A novel polymer monolith based on the dicationic crosslinker 3,3'-(hexane-1,6-diyl)bis(1-vinylimidazolium) bromide, the monomer 1-vinylimidazole and a ternary porogen mixture (1-propanol, decan-1-ol and water) was developed and optimized for capillary electrochromatography. This aim was accomplished by adjusting the composition of individual constituents in the polymerization mixture and monitored based on several relevant parameters (e.g. pore structure by scanning electron microscopy, generation of electroosmotic flow, or permeability of material). The ultimately selected composition yielded a monolithic phase which excellently resolved six methylxanthines (including caffeine, theobromine and theophylline) in 15 min. Key requirements concerning the utilized buffer were an acidic pH of 3 and the addition of 50% acetonitrile; additionally, a negative voltage (-25 kV) had to be applied during analyses. The proposed separation mechanism was mixed mode, i.e. the combination of electrostatic repulsion and hydrophobic interaction. Monolith fabrication as well as separation efficiency were found to be highly repeatable, the material was mechanically stable and useable for at least 150 injections. Thus the presented stationary phase is definitely a very promising option for CEC.


Assuntos
Eletrocromatografia Capilar/métodos , Reagentes de Ligações Cruzadas/síntese química , Imidazóis/síntese química , Polivinil/síntese química , Xantinas/análise , Estrutura Molecular , Polimerização , Pressão , Solventes/química , Fatores de Tempo
12.
Asian J Transfus Sci ; 13(2): 110-114, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31896917

RESUMO

INTRODUCTION: Rh discrepancies produced by partial and weak D phenotypes are a problem during routine testing. Some blood units with weak and partial D expression may be missed by serology. Overcoming the limitations of serology can be achieved by molecular typing. Our objective was to evaluate currently used serologic methods with the molecular analysis in solving discrepant results of weak and partial D (Rh) in South Egypt. PATIENTS AND METHODS: Fifty blood donor and patient samples with undetermined D phenotype were subjected to serology to define their phenotype using identification (ID)-Card "ID-partial RhD typing set" using six monoclonal anti-D panels, followed by molecular typing using polymerase chain reaction sequence-specific primer kit. RESULTS: Molecular typing confirmed most of the serology results; two samples previously resolved as partial D Type 3 and DFR by serological methods were clarified by molecular techniques - one sample as weak Type 4 and the other sample as weak Type 3. Among the weak D alleles found in our study, Type 4 was the most common, with a frequency of 20%, followed by Type 3 (14%), Type 1 (8%), Type 2 (6%), and finally, Type 5 with a frequency of 3%. The most common types of partial D were partial D Type D5 (14%) and Type D3 (10%). CONCLUSION: Our study identified D variants (weak D and partial D categories) of the antigen D and determined the frequency and composition of partial D and weak D alleles in our population. Molecular typing also confirmed most of the results obtained from serological methods.

13.
Asian Pac J Cancer Prev ; 19(9): 2535-2543, 2018 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-30256048

RESUMO

Background: P-glycoprotein (P-gp), a membrane transporter encoded by the multidrug resistance-1 (MDR1) gene, influences pharmacokinetics and metabolism of anticancer drugs and contributes to multidrug resistance phenotype in acute lymphoblastic leukemia (ALL). Genetic variation ofMDR1 in ALL patients is increasingly recognized as a factor influencing response to treatment. Aim: To investigate the possible role of MDR-1 gene polymorphisms (C3435T, C1236T and C4125A) as risk factors for the development and clinical outcome of ALL in Egyptian children. Materials and Methods: Genotyping of MDR-1 C3435T, C1236T and C4125A single nucleotide polymorphisms (SNPs) was accomplished using a polymerase chain reaction­restriction fragment length polymorphism (RFLP-PCR) assay with 120 childhood ALL patients and 100 healthy controls. Results: Homozygous T with the C3435T SNP showed a protective effect as compared to homozygous C (OR=0.748) while heterozygous CT correlated with a poor outcome (high risk, drug unresponsiveness, relapse and high percentage of death). Additionally, the T allele of the C1236T SNP showed a significant relation with ALL risk (OR=1.6). However, there were no significant differences in the genotype and allele frequencies of MDR-1 SNPs between patients and controls. Only one genotype (CC) and one allele of MDR-1 (C4125A) were seen. Neither CA/AA genotypes nor A alleles were present in ALL patients and normal controls. TC was the predominant haplotype in both groups, while CT proved to be minor. The cumulative incidence of relapse was higher with the CC genotype of C1236T as compared with TT. Conclusion: From our preliminary data, the CT genotype of C3435T is associated with a poor ALL outcome while the CC genotype of C1236T is related with an increased incidence of relapse. Although our results provide assistance for oncologist choice of individual therapeutic strategy taking the patient genetic repertoire into consideration, further investigations with larger sample size should be conducted to validate our results.


Assuntos
Biomarcadores Tumorais/genética , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Subfamília B de Transportador de Cassetes de Ligação de ATP/genética , Adolescente , Estudos de Casos e Controles , Criança , Pré-Escolar , Egito , Feminino , Seguimentos , Genótipo , Humanos , Lactente , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Prognóstico
14.
Ann Hepatol ; 16(4): 574-583, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28611261

RESUMO

BACKGROUND AND AIMS: Portal vein thrombosis (PVT) is a critical complication in cirrhotic patients. We explored the role of the activated factor VII-antithrombin (FVIIa-AT) complex and enhanced monocytic tissue factor (TF) expression in the development and prediction of non-neoplastic PVT in cirrhotic patients. MATERIAL AND METHODS: A total of 30 HCV-cirrhosis patients were included in our study. They were compared to 35 cirrhotic patients without PVT, 15 non-cirrhotic patients with PVT, and 15 healthy controls. The plasma level of the FVIIa-AT complexes was analyzed by ELISA. MIF CD142, CD86, and HLA-DR on monocytes (CD14) were determined by flow cytometry. RESULTS: Compared with cirrhotic patients without PVT, cirrhotic patients with PVT had comparable plasma values of FVIIa, AT, and the FVIIa-AT complex. However, they had significantly lower values compared to non-cirrhotic patients with PVT and healthy controls. Cirrhotic patients with PVT had increased monocytic TF expression (MIF CD142) compared to non-PVT cirrhotic patients and healthy controls [86.5 (93.5) vs. 18 (32.0) and 11.0 (6.0), respectively; p < 0.001 for each]. However, cirrhosis PVT could not be distinguished from non-cirrhosis PVT. The area under the ROC curve of MIF CD142 was 0.759 (0.641- 0.876; p = 0.000) at an optimal cut-off value of 45, which yielded a sensitivity of 60% and a specificity of 77.1%, as well as a PPV and NPV of 69.2% for each. CONCLUSION: Enhanced expression of monocytic TF may have a role in the development and prediction of non-neoplastic PVT in HCV-cirrhosis patients. Large multicenter studies are necessary to validate our results.


Assuntos
Antitrombinas/análise , Coagulação Sanguínea , Fator VIIa/análise , Hepatite C/complicações , Cirrose Hepática/sangue , Veia Porta , Tromboplastina/análise , Trombose Venosa/sangue , Adulto , Área Sob a Curva , Biomarcadores/sangue , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Feminino , Hepatite C/sangue , Hepatite C/diagnóstico , Hepatite C/imunologia , Humanos , Cirrose Hepática/diagnóstico , Cirrose Hepática/imunologia , Cirrose Hepática/virologia , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Complexos Multiproteicos , Análise Multivariada , Veia Porta/diagnóstico por imagem , Valor Preditivo dos Testes , Estudos Prospectivos , Curva ROC , Fatores de Risco , Trombose Venosa/diagnóstico , Trombose Venosa/imunologia , Trombose Venosa/virologia , Adulto Jovem
15.
Anal Chim Acta ; 963: 136-142, 2017 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-28335967

RESUMO

A methacrylate based monolith, containing the innovative zwitterionic monomer (3-allyl-1-imidazol)propane sulfonate, was prepared in 100 µm I.D. silica capillaries by UV initiated photo-polymerization. Composition of the porogen, i.e. a mixture of 1-propanol, 1,4 butanediol and water, was of great importance to obtain a homogeneous monolith with satisfactory permeability and good electrochromatographic performance. Morphology of the stationary phase was studied in Scanning Electron Microscopy and IR experiments, which revealed a good attachment to the capillary wall, flowthrough-pores in the range of 0.5-2 µm, and a continuous monolithic structure. The developed material was well suited for the analysis of six common phenolic acids (salicylic, cinnamic, syringic, rosmarinic, caffeic and chlorogenic acid) by CEC. Their separation was possible in less than 8 min with a mobile phase comprising a 12 mM aqueous ammonium acetate solution with pH 8.5 and acetonitrile, at an applied voltage of - 20 kV. The developed method was validated (R2 ≥ 0.995; LOD ≤ 3.9 µg mL-1, except for salicylic acid; recovery rates from 94 to 104%) and successfully used for the determination of phenolic acids in Coffea arabica samples. All of them contained cinnamic, syringic and caffeic acid, however only in unroasted coffee beans chlorogenic acid (0.06%) was found. The quantitative results were in good agreement to reported literature data.


Assuntos
Eletrocromatografia Capilar/métodos , Café/química , Hidroxibenzoatos/isolamento & purificação , Imidazóis/química , Extratos Vegetais/química
16.
Artigo em Inglês | MEDLINE | ID: mdl-28053672

RESUMO

BACKGROUND: Serotonin (5-HT) improves insulin sensitivity and glucose metabolism, however, the underlying molecular mechanism has remained elusive. Previous studies suggest that 5-HT can activate intracellular small GTPases directly by covalent binding, a process termed serotonylation. Activated small GTPases have been associated with increased GLUT4 translocation to the cell membrane. Therefore, we investigated whether serotonylation of small GTPases may be involved in improving Insulin sensitivity and glucose metabolism. METHODS: Using fully differentiated L6 rat skeletal muscle cells, we studied the effect of 5-HT in the absence or presence of insulin on glycogen synthesis, glucose uptake and GLUT4 translocation. To prove our L6 model we additionally performed preliminary experiments in C2C12 murine skeletal muscle cells. RESULTS: Incubation with 5-HT led to an increase in deoxyglucose uptake in a concentration-dependent fashion. Accordingly, GLUT4 translocation to the cell membrane and glycogen content were increased. These effects of 5-HT on Glucose metabolism could be augmented by co-incubation with insulin and blunted by co incubation of 5-HT with monodansylcadaverine, an inhibitor of protein serotonylation. In accordance with this observation, incubation with 5-HT resulted in serotonylation of a protein with a molecular weight of approximately 25 kDa. We identified this protein as the small GTPase Rab4, the activity of which has been shown to be stimulated by both insulin signalling and serotonylation. CONCLUSION: Our data suggest that 5-HT elicits its beneficial effects on Glucose metabolism through serotonylation of Rab4, which likely represents the converging point between the insulin and the 5-HT signalling cascades.

17.
Artigo em Inglês | MEDLINE | ID: mdl-27785340

RESUMO

BACKGROUND AND OBJECTIVE: Human umbilical cord blood (UCB) cells and bone marrow mesenchymal stem cells (BM-MSCs) have numerous advantages as grafts for cell transplantation. We hypothesized differing impacts of human UCB cells and rat BM-MSCs on reversal of hepatic injury and revival of liver function in carbon tetrachloride (CCl4)-induced liver fibrosis. METHODS: Forty rats were divided into 4 groups; control group, CCl4 group, CCl4/CD34+ group and CCl4/BM-MSCs group. Blood samples were driven from rats at 4, 8 and 12 weeks to measure serum concentration of albumin and alanine aminotransferase (ALT). Quantitative expression of collagen Iα, TGF-ß, α-SMA, albumin, MMP-2, MMP-9 and TNF-α were assessed by polymerase chain reaction. Histopathological examination of the liver tissue was performed. GFP labeled cells were detected in groups injected with stem cells. RESULTS: Regarding liver function, CD34+ were more efficient than BM-MSCs in elevating albumin (P<0.05) and reducing ALT (P<0.05) concentrations. Concerning gene expression, CD34+ were more effective than BM-MSCs in reducing gene expressions of collagen Iα (P<0.01), TGF-ß1 (P<0.01) and α-SMA (P<0.01). Both CD34+ and BM-MSCs have the same efficacy in reducing TNF-α (P<0.001 and P<0.01, respectively). Furthermore, CD34+ were more valuable than BM-MSCs in increasing gene expression of albumin (P<0.05) and MMP-9 (P<0.01). CONCLUSION: Taken together; human UCB CD34+ stem cells were more efficient in improvement of experimental liver injury than BM-MSCs. This study highlighted an important role of human UCB CD34+ stem cells in liver fibrosis therapy.

18.
Hum Immunol ; 77(10): 937-943, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27343994

RESUMO

Several evidences suggest that DN T cells, IL23 and IL6 play a role in the pathogenesis of SLE. This study aimed to evaluate the frequency of DN T cells in SLE patients and the relation to their activity also to assess the possible role of IL6 and IL23 on DN T cells. Thirty patients with SLE and sixteen healthy blood donor females were enrolled. There was a significant increase in DN T cells in patients than controls (P=0.001). These cells had a significant positive correlation with SLEDAI (r=0.486, P=0.006). DN T cells from SLE patient samples were expanded when stimulated in vitro with RhIL6 or RhIL23 in patients than controls. Furthermore, anti ds-DNA level was found to be increased in supernatant of PBMCs when stimulated by these cytokines in different concentrations. Our findings suggest that IL6 and IL23 may play role in SLE pathogenesis through their effect on DN T cells and anti ds-DNA.


Assuntos
Interleucina-23/metabolismo , Interleucina-6/metabolismo , Lúpus Eritematoso Sistêmico/imunologia , Linfócitos T/imunologia , Adolescente , Adulto , Anticorpos Antinucleares/metabolismo , Antígenos CD4/metabolismo , Antígenos CD8/metabolismo , Estudos de Casos e Controles , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Progressão da Doença , Feminino , Humanos , Ativação Linfocitária , Adulto Jovem
19.
Oncol Lett ; 11(1): 823-830, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26870291

RESUMO

A non-invasive marker is required for the diagnosis and follow-up of patients with bladder cancer. The aim of the current study was to evaluate the potential prognostic significance of serum osteoprotegerin (OPG), p53 protein and urine telomerase in patients with bladder cancer. For all patients, serum levels of OPG and p53 protein were determined using enzyme-linked immunosorbent assay (ELISA), and urine telomerase was assessed using a polymerase chain reaction ELISA technique. Patients were assigned into group 1 (cystectomy and adjuvant radiotherapy) or group 2 (transurethral resection and chemoradiotherapy). The results revealed that serum OPG and p53, and urine telomerase levels were significantly higher in bladder cancer patients compared with in healthy individuals (P<0.0001). High serum OPG was associated with significantly lower overall survival and disease-free survival rates (both P=0.001), and was correlated with advanced tumor stages (P<0.0001), high tumor grades (P<0.0001) and the occurrence of disease relapse (P=0.001). Serum p53 and urine telomerase did not demonstrate prognostic significance. These findings indicate that serum OPG level may be used as a diagnostic tool and a prognostic variable for patients with muscle invasive bladder cancer. Future trials are required to elucidate its therapeutic role in such patients.

20.
Food Chem ; 194: 149-55, 2016 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-26471538

RESUMO

A simple solid-phase extraction method for the enrichment of 5 bisphenol derivatives using hexagonal boron nitride (BN) was developed. BN was applied to concentrate bisphenol derivatives in spiked water samples and the compounds were analyzed using HPLC coupled to fluorescence detection. The effect of pH and organic solvents on the extraction efficiency was investigated. An enrichment factor up to 100 was achieved without evaporation and reconstitution. The developed method was applied for the determination of bisphenol A migrated from some polycarbonate plastic products. Furthermore, bisphenol derivatives were analyzed in spiked and non-spiked canned food and beverages. None of the analyzed samples exceeded the migration limit set by the European Union of 0.6mg/kg food. The method showed good recovery rates ranging from 80% to 110%. Validation of the method was performed in terms of accuracy and precision. The applied method is robust, fast, efficient and easily adaptable to different analytical problems.


Assuntos
Compostos Benzidrílicos/química , Compostos de Boro/química , Cromatografia Líquida de Alta Pressão/métodos , Fenóis/química , Extração em Fase Sólida/métodos
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