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1.
Ann Lab Med ; 42(2): 141-149, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635607

RESUMO

Standardization of cell-free DNA (cfDNA) testing processes is necessary to obtain clinically reliable results. The pre-analytical phase of cfDNA testing greatly influences the results because of the low proportion and stability of circulating tumor DNA (ctDNA). In this review, we provide evidence-based clinical practice guidelines for pre-analytical phase procedures of plasma epidermal growth factor receptor gene (EGFR) variant testing. Specific recommendations for pre-analytical procedures were proposed based on evidence from the literature and our experimental data. Standardization of pre-analytical procedures can improve the analytical performance of cfDNA testing.


Assuntos
Ácidos Nucleicos Livres , DNA Tumoral Circulante , Receptores ErbB/genética , Humanos
2.
Food Chem ; 367: 130620, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34343812

RESUMO

Artichokes are a rich source of (poly)phenols, mainly caffeoylquinic acids, but little is known about their bioavailability from this source. This study investigated the absorption, metabolism and excretion of (poly)phenols after sous-vide artichoke consumption (5776 µmol of (poly)phenols) by healthy volunteers. Seventy-six (poly)phenol metabolites were identified by UHPLC-MS/MS using authentic standards, including acyl-quinic acids plus C6-C3, C6-C1, C6-C2, C6-C1-N, C6-C0 metabolites, and their phase-II conjugates. The major metabolites were 3'-methoxy-4'-hydroxycinnamic acid, 3'-methoxycinnamic acid-4'-sulfate, and 4'-hydroxycinnamic acid-3'-sulfate, which appeared early in plasma (Tmax < 4 h); plus 3-(3'-methoxy-4'-hydroxyphenyl)propanoic acid, 3-(4'-methoxyphenyl)propanoic acid-3'-glucuronide, 3-(3'-hydroxyphenyl) propanoic acid and hippuric acids, which appeared later (Tmax > 6 h). The 24 h urinary recovery averaged 8.9% (molar basis) of the (poly)phenols consumed. Hepatic beta-oxidation of 3',4'-dihydroxycinnamic acid and methylated conjugates occurred, but was limited (<0.04%). 3'-Methylation exceeded 4'-methylation and interindividual variability was high, especially for gut microbial metabolites (up to 168-fold).


Assuntos
Cynara scolymus , Disponibilidade Biológica , Humanos , Metaboloma , Fenóis , Polifenóis , Espectrometria de Massas em Tandem
3.
Ann Lab Med ; 42(1): 54-62, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34374349

RESUMO

Background: Associations between IgA nephropathy (IgAN) and HLA-DRB1 and -DQB1 alleles have been reported in several ethnic groups. We investigated the association of HLA-DRB1 and -DQB1 alleles with the predisposition for IgAN and disease progression to end-stage kidney disease (ESKD) in Korean patients. Methods: We analyzed HLA-DRB1 and -DQB1 genotypes in 399 IgAN patients between January 2000 and January 2019 using a LIFECODES sequence-specific oligonucleotide (SSO) typing kit (Immucor, Stamford, CT, USA) or a LABType SSO Typing Test (One Lambda, Canoga Park, CA, USA). Alleles with a significant difference in two-digit resolution were further analyzed using in-house sequence-based typing and sequence-specific primer PCR. As controls, 613 healthy hematopoietic stem cell donors were included. Kidney survival was analyzed in 281 IgAN patients with available clinical and laboratory data using Cox regression analysis. Where needed, P-values were adjusted using Bonferroni correction. Results: The allele frequencies of HLA-DRB1*04:05 (corrected P [Pc]<0.001), -DQB1 *04:01 (Pc=0.048), and -DQB1*03:02 (Pc=0.021) were significantly higher in IgAN patients than in controls, whereas those of HLA-DRB1*07:01, -DRB1*15:01, -DQB1*02:02, and -DQB1*06:02 (Pc<0.001 for all) were significantly lower in IgAN patients than in controls. The allele frequency of HLA-DQB1*05:03 (Pc=0.016) was significantly lower in the ESKD group than in the non-ESKD group; however, there was no significant difference for ESKD progression between these groups. Conclusions: We report novel associations of HLA-DRB1*15:01, DQB1*02:02, -DQB1*03:02, and -DQB1*04:01 with IgAN. Further studies of HLA alleles associated with IgAN progression in a larger cohort and in various ethnic groups are needed.


Assuntos
Glomerulonefrite por IGA , Alelos , Predisposição Genética para Doença , Glomerulonefrite por IGA/diagnóstico , Glomerulonefrite por IGA/genética , Cadeias HLA-DRB1/genética , Teste de Histocompatibilidade , Humanos , República da Coreia
4.
Ann Lab Med ; 42(1): 63-70, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34374350

RESUMO

Background: Recently, two fully automated immunoassays for antinuclear antibody (ANA) screening were introduced: EliA CTD Screen (Thermo Fisher Scientific, Freiburg, Germany) and QUANTA Flash CTD Screen Plus (Inova Diagnostics, San Diego, USA). We evaluated their clinical performance in comparison with the indirect immunofluorescence assay (IIFA) and analyzed samples with discrepant results. Methods: In total, 406 serum samples (206 from patients undergoing routine checkups and 200 from rheumatology clinic patients) were assayed using EliA, QUANTA Flash, and IIFA. We evaluated assay concordance and agreement and confirmed the presence of anti-extractable nuclear antigen (ENA) antibodies in samples with discrepant automated immunoassay and IIFA results. Additionally, we compared the clinical performance of each assay in diagnosing ANA-associated rheumatic disease (AARD) and adjusted the cut-off values. Results: In rheumatology clinic samples, the concordance and agreement were 91.5% and strong between EliA and QUANTA Flash, 79.0% and weak between EliA and IIFA, and 80.5% and moderate between QUANTA Flash and IIFA, respectively. In automated immunoassay-positive, IIFA-negative samples (N=15), all anti-ENA antibodies detected (6/15) were anti-Sjögren's syndrome antigen A/Ro (Ro60) antibodies. The automated immunoassays and IIFA showed high accuracy for diagnosing AARD, and adjusted cut-off values improved their sensitivities (EliA with 0.56 ratio, 82.9% sensitivity; QUANTA Flash with 9.7 chemiluminescent units, 87.8% sensitivity). Conclusions: The two automated immunoassays showed reliable performance compared with IIFA and can be efficiently used with the IIFA in clinical immunology laboratories. Clinical cut-off values can be adjusted according to the workflow in each laboratory.


Assuntos
Anticorpos Antinucleares , Programas de Rastreamento , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Imunoensaio , Sensibilidade e Especificidade
5.
Talanta ; 236: 122806, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635208

RESUMO

Glucagon-like peptide (GLP)-1 and the glucose-dependent insulinotropic peptide (GIP) are incretin hormones that regulate the nutrient-stimulated insulin secretion from pancreatic ß-cells. Their low plasma concentrations and rapid clearance pose certain methodological challenges for their detection and quantification. The currently available immunomediated techniques to monitor these hormones overestimate, to some extent, their actual concentration. Hence, the present study is aimed at developing a robust and reliable methodology for the identification and quantification of active and inactive forms of the incretins GLP-1 and GIP, in human plasma, by UHPLC-ESI-QqQ-MS/MS. A comparative study of different SPE cartridges was carried out, being identified OASIS HLB as the most efficient one, with recoveries up to 80%. The method provides adequate linearity, from 4.88 to 1250 nM, and low intervals of LOD and LOQ for each analyte (ranges from 0.01 to 3.42 nM and from 0.10 to 34.17 nM, respectively). The methodology described was validated upon a clinical trial with overweight subjects (n = 20) (ClinicalTrials.gov NCT04016337), showing the capacity of the newly developed methodology to detect the augment of the plasma concentration of both GLP-17-36 and GLP-19-36 between 30 and 60 min after the consumption of a sucrose sweetened fruit-based beverage, while the plasma concentration of GIP remained in levels lower than the LOQ. The proposed methodology provides further insights into the mechanisms of action of bioactive compounds and food components in the frame of the glycemic control and would contribute to the assessment of the efficacy of antidiabetic drugs.


Assuntos
Polipeptídeo Inibidor Gástrico , Incretinas , Cromatografia Líquida de Alta Pressão , Peptídeo 1 Semelhante ao Glucagon , Humanos , Espectrometria de Massas em Tandem
6.
Talanta ; 236: 122827, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635217

RESUMO

Cryptococcal meningitis (CM) is a global threat with significant attributable morbidity and mortality. Information on microfluidic detection for CM diagnosis is still limited. We developed a multifunctional microfluidic module that integrated the pathogen enrichment and on-chip nucleic acid extraction. The module adopted a simple filtration membrane to effectively capture Cryptococcus cells and simplify the process, and combined lyticase digestion, alkaline lysis and heating methods to optimize the strategy to achieve nucleic acid extraction. The entire process was operated in the module, which reduced the exposure risk of directly processing cryptococcal samples. A portable one-pot lyophilized LAMP reagent bead with no temperature limit was developed, which improved the flexibility of operation. This module did not require any additional instrument, and is promising to develop a simple, rapid, and efficient approach to realize the "sample in and answer out" detection of real CSF samples. This microfluidic module had practical prospects and is expected to replace LFA for efficacy evaluation and follow-up in the middle and late stages of CM treatment, and could be used as an auxiliary method to confirm cases with questionable LFA results in the early diagnosis of CM.


Assuntos
Meningite Criptocócica , Ácidos Nucleicos , Humanos , Meningite Criptocócica/diagnóstico , Microfluídica , Análise de Sequência com Séries de Oligonucleotídeos
7.
Talanta ; 236: 122830, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635220

RESUMO

A sensitive biosensor that can be used for the determination of matrix metalloproteinase 2 (MMP-2) was proposed. The biosensor was developed by using an excellent self-enhanced nanocomposites as an illuminant and a peptide as a recognition element. For the electrostatic attraction between Ru(bpy)32+ and nitrogen-doped graphene quantum dots (NGQDs), the self-enhanced electrochemiluminescence (ECL) nanocomposites of NGQDs-Ru(bpy)32+-doped silica nanoparticles (NGQDs-Ru@SiO2) were synthesized through a simple sol-gel process. Then, a specific peptide (labeled sulfhydryl) was combined with the self-enhanced ECL nanocomposites (carboxyl in NGQDs) via acylation reaction to obtain the peptide-NGQDs-Ru@SiO2 nanoprobe, which was fabricated onto the gold electrode surface via Au-S bond. The peptide of the ECL nanoprobe was exposed to cleavage in the presence of MMP-2, which caused the signal substance to move farther away from the electrode, leading to a decrease of the ECL signal. The proposed NGQDs-Ru@SiO2-labeled peptide ECL biosensor displayed a lower detection limit of 6.5 pg mL-1 than those of reported ECL methods. The proposed biosensor provided an outlook for future applications in other disease-associated biomarkers.


Assuntos
Técnicas Biossensoriais , Grafite , Neoplasias , Pontos Quânticos , Biomarcadores Tumorais , Técnicas Eletroquímicas , Humanos , Medições Luminescentes , Metaloproteinase 2 da Matriz , Nitrogênio , Dióxido de Silício
8.
Talanta ; 236: 122831, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635221

RESUMO

In this study, a novel type of covalent organic framework (COF) material rich in boronic acid sites was prepared through post-synthesis modification (TbBD@PEI@Au@4-MPBA). The surface of COF material had abundant carboxylic acid groups, which could bind a large amount of polyethyleneimine (PEI) through electrostatic interaction. At the same time, the amino groups on the PEI can be grafted with Au nanoparticles (Au NPs) in situ, and then 4-mercaptophenylboronic acid (4-MPBA) was modified by the reaction of Au and sulfhydryl groups. The massive grafting of boronic acid groups made the material's enrichment effect on glycopeptides expected. The results of experiments indicated that the composite material has high sensitivity (5 amol µL-1) and selectivity (1:1000). In addition, the material has outstanding stability and reusability, with a load capacity of about 100 mg g-1 and a recovery of 99.3 ± 2.2%. What's more, after enriched by TbBD@PEI@Au@4-MPBA, 56 endogenous glycopeptides from fresh human saliva were detected by MALDI-TOF MS, 56 unique glycopeptides corresponding to 31 glycoproteins from human saliva and 513 unique glycopeptides corresponding to 208 glycoproteins from serum of throat cancer patient were detected by nano-LC-MS/MS, respectively, which was expected to be applied to glycoproteomics research.


Assuntos
Nanopartículas Metálicas , Estruturas Metalorgânicas , Glicopeptídeos , Ouro , Humanos , Interações Hidrofóbicas e Hidrofílicas , Saliva , Espectrometria de Massas em Tandem
9.
Talanta ; 236: 122839, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635229

RESUMO

Single-cell analysis of proteins is critical to gain precise information regarding the mechanisms that dictate the heterogeneity in cellular phenotypes and their differential response to internal and external stimuli. However, tools that allow sensitive and easy measurement of proteins in individual cells are still limited. The emerging semiconductor-based bioelectronics may provide a new approach to overcome the challenges in this field, however its utility in single-cell protein analysis has not been explored. In this study, we investigated multiple protein detection in single cells by MoS2 field effect transistors (MoS2-FETs) modified with specific biological probes. First, ß-actin antibody was connected to the surface of MoS2-FETs by covalent bonds, and the fabricated device was tested using ß-actin solution with concentrations from 10-9 to 10-3 µg/µL. Next, we examined the application of MoS2-FET for protein analysis in complex biological samples, and the device showed electrical signal response to human embryonic kidney cell line HEK293T in a dose-dependent manner. Furthermore, we applied this method to analyze individual liver cancer MHCC-97L cells, targeting four cellular proteins, including ß-actin, epidermal growth factor receptor, sirtuin-2, and glyceraldehyde-3-phosphate dehydrogenase. The devices modified with corresponding probes could identify the target proteins and showed cell number-dependent responses. As a proof of principle, we demonstrated sensitive and multiplexed detection of proteins in single cells using MoS2-FETs. The biosensor and this detection method are cost-efficient and user-friendly with broad application prospects in biological studies and clinical diagnosis.


Assuntos
Técnicas Biossensoriais , Molibdênio , Células HEK293 , Humanos , Proteínas , Semicondutores
10.
Talanta ; 236: 122843, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635233

RESUMO

In our study, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) is proposed as a novel tool, which can be applied to analyze lipids in urine samples. For this reason, the main aim of the study was to develop and optimize the preparation protocol for urine samples in lipidomics, using urine samples obtained from patients with diagnosed cancer and non-cancer controls. Several conditions like extraction method and types of matrices were evaluated. For this purpose, two methods for the extraction of lipids, namely modified Folch and Bligh & Dyer were employed. Furthermore, two types of matrices (α-cyano-4-hydroxycinnamic acid (HCCA) and 2,5-dihydroxybenzoic acid (DHB)) for the separation of lipids into individual components was tested. The results of this study can serve as an essential source for the selection of appropriate extraction methods and the appropriate choice of a matrix for the purification and identification of a particular class of lipid in human biological fluids. Based on it, Bligh & Dyer method associated with the usage of HCCA matrix was found to be the most effective for lipidomics using MALDI-TOF/MS. The optimized method was applied to compare the lipid profile of 139 urine samples collected from both healthy individuals and patients with prostate cancer. The tandem spectroscopic analysis allowed to identify lysophosphatidylcholine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and triacylglycerols in urine samples. Finally, MALDI-TOF/MS analysis enabled to discriminate between the two tested groups (healthy individuals and patients with prostate cancer). A preliminary statistical model suggested that classification accuracy ranging from 83.3 to100.0% may be achieved by using pre-selected MS signals.


Assuntos
Fosfatidilcolinas , Neoplasias da Próstata , Humanos , Masculino , Neoplasias da Próstata/diagnóstico , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Triglicerídeos
11.
Talanta ; 236: 122867, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635249

RESUMO

Carcinoembryonic antigen (CEA) is one of the most widely used tumor marker around the world, it mainly used for gastrointestinal cancers, especially in colorectal malignancy. At present, the detection methods of CEA are mostly based on antigen-antibody binding, whereas these methods were limited by the high costs and long waiting times in massive population tumor screening. During the experiments, we interestingly found that the fluorescence signal would be dramatically altered when the secondary structure of fluorescent modified guanine-rich DNA changed. Then we explored the reasons and established a new method for CEA detection, this method brings a simple, fast and cheap sensing platform for detection of biomarkers. It has great potential in screening of tumors among the group and is expected to provide prospective effects for tumor treatment.


Assuntos
Aptâmeros de Nucleotídeos , Neoplasias Colorretais , Antígeno Carcinoembrionário , Neoplasias Colorretais/diagnóstico , Fluorescência , Guanina , Humanos
12.
Talanta ; 236: 122879, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635259

RESUMO

We present a sensitive label-free surface enhanced Raman spectroscopy (SERS) method for the discrimination between the recombinant and endogenous human Erythropoietin (EPO) isoforms. The proposed methodology comprises a lectin-functionalised extractor chip for the extraction of the recombinant human EPO (rhuEPO) and the endogenous EPO (enEPO) from blood plasma. The disulfide bond molecular structure of the purified isoforms was modified to chemisorb the biomolecules onto a SERS substrate in a unified orientation, thus maximizing the reproducibility and sensitivity of the SERS measurements. The acquired SERS spectra of the EPO isoforms showed diagnostic Raman bands that allowed for the discrimination between rhuEPO and enEPO. The method was also used for the SERS quantification of rhuEPO and enEPO down to 0.1 pM and 0.5 pM, respectively. The SERS determination of the protein isoforms was cross validated against ELISA. The new SERS method has strong potential for the rapid screening of rhuEPO doping in athletes and for the therapeutic drug monitoring of rhuEPO treatment in cancer patients.


Assuntos
Eritropoetina , Análise Espectral Raman , Humanos , Isoformas de Proteínas , Proteínas Recombinantes , Reprodutibilidade dos Testes
13.
Talanta ; 236: 122886, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635266

RESUMO

Rheumatoid arthritis (RA), an autoimmune and chronic inflammatory disorder, is an incurable disease. We developed a peptide-based electrochemical sensor using electrochemical impedance spectroscopy that can be used to detect autoantibodies for RA diagnostics. We first validated that the developed peptide showed high sensitivity and could compliment the current gold standard method of an anti-cyclic citrullinated peptide antibody (anti-CCP) ELISA. The developed peptide can be modified on the nanogold surface of the working electrode of sensing chips through the method of a self-assembling monolayer. The sensing process was first optimized using a positive control cohort and a healthy control cohort. Subsequently, 10 clinically confirmed samples from RA patients and five healthy control samples were used to find the threshold value of the impedance between RA and healthy subjects. Furthermore, 10 clinically confirmed samples but with low values of anti-CCP autoantibodies were used to evaluate the sensitivity of the present method compared to the conventional method. The proposed method showed better sensitivity than the current conventional anti-CCP ELISA method.


Assuntos
Artrite Reumatoide , Artrite Reumatoide/diagnóstico , Espectroscopia Dielétrica , Impedância Elétrica , Ensaio de Imunoadsorção Enzimática , Humanos , Peptídeos
14.
Talanta ; 236: 122899, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635272

RESUMO

A real-time quartz crystal microbalance (QCM) cytosensor was first developed for dynamical and noninvasive monitoring of cell viscoelasticity for evaluation of apoptosis degree. In this work, human breast cancer cells MCF-7 and MDA-MB-231 were employed as cell model and respectively captured on the surface of QCM electrode modified with mercaptosuccinic acid and poly-l-lysine. Cell viscoelasticity was measured dynamically by real-time monitoring energy dissipation with QCM, and the dynamic diagram of the energy dissipation of MDA-MB-231 cells treated with curcumin was first obtained. The results displayed that the changes of energy dissipation in MDA-MB-231 cells and MCF-7 cells were 8.81 × 10-6 and 5.29 × 10-6, particularly due to the difference in cell viscoelasticity. Furthermore, curcumin was used to induce cell apoptosis and suppress energy dissipation of MDA-MB-231 cells. Combining apoptosis assay with QCM measurement, the results revealed good linear relationship between cell viscoelasticity inhibition and apoptosis rate with correlation coefficient R = 0.9908. The QCM cytosensor could rapidly, accurately, dynamically, and noninvasively monitor the changes of cell viscoelasticity for evaluation of apoptosis degree in MDA-MB-231 cells. The study established a new model for cell apoptosis assessment, facilitating understanding of the mechanisms of cell apoptosis on the aspect of mechanical properties.


Assuntos
Técnicas Biossensoriais , Neoplasias da Mama , Curcumina , Apoptose , Neoplasias da Mama/tratamento farmacológico , Curcumina/farmacologia , Feminino , Humanos , Viscosidade
15.
Ann Lab Med ; 42(2): 188-195, 2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635612

RESUMO

Background: Fusobacterium species are obligately anaerobic, gram-negative bacilli. Especially, F. nucleatum and F. necrophorum are highly relevant human pathogens. We investigated clinical differences in patients infected with Fusobacterium spp. and determined the antimicrobial susceptibility of Fusobacterium isolates. Methods: We collected clinical data of 86 patients from whom Fusobacterium spp. were isolated from clinical specimens at a tertiary-care hospital in Korea between 2003 and 2020. In total, 76 non-duplicated Fusobacterium isolates were selected for antimicrobial susceptibility testing by the agar dilution method, according to the Clinical and Laboratory Standards Institute guidelines (M11-A9). Results: F. nucleatum was most frequently isolated from blood cultures and was associated with hematologic malignancy, whereas F. necrophorum was mostly prevalent in head and neck infections. Anti-anaerobic agents were more commonly used to treat F. nucleatum and F. varium infections than to treat F. necrophorum infections. We observed no significant difference in mortality between patients infected with these species. All F. nucleatum and F. necrophorum isolates were susceptible to the antimicrobial agents tested. F. varium was resistant to clindamycin (48%) and moxifloxacin (24%), and F. mortiferum was resistant to penicillin G (22%) and ceftriaxone (67%). ß-Lactamase activity was not detected. Conclusions: Despite the clinical differences among patients with clinically important Fusobacterium infections, there was no significant difference in the mortality rates. Some Fusobacterium spp. were resistant to penicillin G, ceftriaxone, clindamycin, or moxifloxacin. This study may provide clinically relevant data for implementing empirical treatment against Fusobacterium infections.


Assuntos
Anti-Infecciosos , Infecções por Fusobacterium , Fusobacterium , Infecções por Fusobacterium/diagnóstico , Infecções por Fusobacterium/tratamento farmacológico , Humanos , República da Coreia , Centros de Atenção Terciária
16.
Ann Lab Med ; 42(2): 196-202, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635613

RESUMO

Background: Identifying the causal pathogen of encephalitis remains a clinical challenge. A 50-year-old man without a history of neurological disease was referred to our department for the evaluation of an intracranial lesion observed on brain magnetic resonance imaging (MRI) scans, and the pathology results suggested protozoal infection. We identified the species responsible for encephalitis using thymine-adenine (TA) cloning, suitable for routine clinical practice. Methods: We extracted DNA from a paraffin-embedded brain biopsy sample and performed TA cloning using two universal eukaryotic primers targeting the V4-5 and V9 regions of the 18S rRNA gene. The recombinant plasmids were extracted, and the inserted amplicons were identified by Sanger sequencing and a homology search of sequences in the National Center for Biotechnology Information Basic Local Alignment Search Tool. Results: The infection was confirmed to be caused by the free-living amoeba Balamuthia mandrillaris. Two of 41 colonies recombinant with 18S V4-5 primers and 35 of 63 colonies recombinant with the 18S V9 primer contained B. mandrillaris genes; all other colonies contained human genes. Pathogen-specific PCR ruled out Entamoeba histolytica, Naegleria fowleri, Acanthamoeba spp., and Toxoplasma gondii infections. Conclusions: This is the first report of B. mandrillaris-induced encephalitis in Korea based on molecular identification. TA cloning with the 18S rRNA gene is a feasible and affordable diagnostic tool for the detection of infectious agents of unknown etiology.


Assuntos
Balamuthia mandrillaris , Encefalite , Adenina , Balamuthia mandrillaris/genética , Clonagem Molecular , Encefalite/diagnóstico , Eucariotos , Humanos , Masculino , Pessoa de Meia-Idade , Timina
17.
Ann Lab Med ; 42(2): 249-257, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635616

RESUMO

Background: Non-invasive clinical algorithms for the detection of liver fibrosis (LF) can reduce the need for liver biopsy (LB). We explored the implementation of two serum biomarkers, enhanced liver fibrosis (ELF) and Mac-2 binding protein glycosylation isomer (M2BPGi), in clinical algorithms for LF in chronic hepatitis B (CHB) patients. Methods: Two clinical algorithms were applied to 152 CHB patients: (1) transient elastography (TE) followed by biomarkers (TE/ELF and TE/M2GPGi); (2) biomarker test followed by TE (ELF/TE and M2BPGi/TE). Using the cut-off value or index for the detection of advanced LF (TE≥F3; 9.8 in ELF and 3.0 in M2BPGi), LB was expected to be performed in cases with discordant TE and biomarker results. Results: In both algorithms, the expected number of LBs was lower when using M2BPGi than when using ELF (TE/ELF or ELF/TE, 13.2% [N=20]; TE/M2BPGi or M2BPGi/TE, 9.9% [N=15]), although there was no statistical difference (P=0.398). In the TE low-risk group (TE≤F2), the discordance rate was significantly lower in the TE/M2BPGi approach than in the TE/ELF approach (1.5% [2/136] vs. 11.0% [15/136], P=0.002). In the biomarker low-risk group, there was no significant difference between the ELF/TE and M2BPGi/TE approaches (3.9% [5/126] vs. 8.8% [13/147], P=0.118). Conclusions: Both ELF and M2BPGi can be implemented in non-invasive clinical algorithms for assessing LF in CHB patients. Given the lowest possibility of losing advanced LF cases in the low-risk group when using the TE/M2BPGi approach, this combination seems useful in clinical practice.


Assuntos
Hepatite B Crônica , Algoritmos , Biópsia , Glicosilação , Hepatite B Crônica/complicações , Hepatite B Crônica/diagnóstico , Humanos , Cirrose Hepática/diagnóstico
18.
Food Chem ; 368: 130827, 2022 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-34411855

RESUMO

Curcuminoids are known to exert diverse pharmacological effects and used in some pharmaceutical formulations. This study describes the preparation, characterization, and enhancement in the solubility and anticancer activity of a curcuminoids-rich extract (CRE) using a ternary inclusion complex system. CRE containing 88.9% w/w curcuminoids was prepared using a 'green' microwave extraction coupled with fractionation on a column of hydrophobic adsorbent resin. The ternary complex consisting of CRE, hydroxylpropyl-ß-cyclodextrin and polyvinylpyrrolidone K30 was prepared using the solvent evaporation method and thoroughly characterized using Fourier-transform infrared spectroscopy, powder X-ray diffractograms, differential scanning calorimetry and scanning electron microscopy. The ternary complex of CRE improved the water-solubility of curcuminoids (up to 70.3 µg/mL) as well as the dissolution rate when compared to those of CRE (0 µg/mL). In addition, the ternary complex exhibited significantly stronger anticancer activity against human lung adenocarcinoma (A-549), human cervical adenocarcinoma (HeLa) and human colon adenocarcinoma (HT-29) cell lines than CRE.


Assuntos
Diarileptanoides , Água , Varredura Diferencial de Calorimetria , Humanos , Extratos Vegetais , Solubilidade , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
19.
Food Chem ; 368: 130839, 2022 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-34419799

RESUMO

Human bitter taste receptor TAS2R14 (T2R14) can widely perceive bitterness, which has always been an issue for people to overcome. This study was aimed at identifying bioactive peptides obtained from Oncorhynchus mykiss nebulin hydrolysates as bitter taste receptor blockers by physicochemical property prediction, molecular docking, and in vitro determination of bitterness intensity using electronic tongue. Exploration of the interaction mechanism of these peptides with T2R14 by molecular docking models indicated that peptides ADM and ADW had high affinities for T2R14 to block the binding of bitter substances into the receptor. Addition of ADM and ADW to quinine caused reduction in bitterness intensity, with IC50 values of 420.32 ± 6.26 µM and 403.29 ± 4.10 µM, respectively. Hydrogen bond interaction and hydrophobic interaction were responsible for manifesting the high affinities of these peptides for the receptor. Residues Thr86, Asp168, and Phe247 may be the key amino acids within the binding site.


Assuntos
Oncorhynchus mykiss , Paladar , Animais , Humanos , Simulação de Acoplamento Molecular , Proteínas Musculares , Oncorhynchus mykiss/genética , Peptídeos/genética , Receptores Acoplados a Proteínas G/genética
20.
Food Chem ; 368: 130871, 2022 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-34438174

RESUMO

This study is the first dynamic simulation of gastrointestinal digestion of cranberry polyphenols [1 g cranberry extract per day (206.2 mg polyphenols) for 18 days]. Samples from the simulated ascending, transverse, and descending colon of the dynamic gastrointestinal simulator simgi® were analyzed. Results showed that 67% of the total cranberry polyphenols were recovered after simulated gastrointestinal digestion. Specifically, benzoic acids, hydroxycinnamic acids, phenylpropionic acids, phenylacetic acids, and simple phenols were identified. Cranberry feeding modified colonic microbiota composition of Enterococcaceae population significantly. However, increments in microbial-derived short-chain fatty acids, particularly in butyric acid, were observed. Finally, the simgi® effluent during cranberry feeding showed significant antiadhesive activity against uropathogenic Escherichia coli (13.7 ± 1.59 % of inhibition). Understanding the role that gut microbiota plays in cranberry metabolism could help to elucidate its interaction with the human body and explain cranberry protective effects against urinary tract infections.


Assuntos
Vaccinium macrocarpon , Bactérias/genética , Digestão , Humanos , Extratos Vegetais/farmacologia , Polifenóis/farmacologia
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