RESUMEN
Neonatal jaundice is a very common disease in newborns and can lead to brain damage or death in severe cases. Phototherapy with light-emitting diode (LED) arrays is widely used as the easiest and fastest way to relieve jaundice in newborns, but it has distinct disadvantages such as loss of water in the patient, damage to the retina, and separation from parents. In this paper, a novel light source-based phototherapy for neonatal jaundice is proposed using a textile-based wearable organic light-emitting diode (OLED) platform that can move flexibly and conform to the curvature of the human body. The soft and flexible textile-based blue OLED platform is designed to have a peak wavelength of 470 nm, suitable for jaundice treatment, and shows performance (>20 µW cm-2 nm- 1 ) suitable for intensive jaundice treatment even at low voltage (<4.0 V). The textile-based OLEDs fabricated in this study exhibit an operating reliability of over 100 h and low-temperature operation (<35 °C). The results of an in vitro jaundice treatment test using a large-area blue OLED confirm that the bilirubin level decreases to 12 mg dL-1 with 3 h of OLED irradiation.
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Ictericia Neonatal , Ictericia , Dispositivos Electrónicos Vestibles , Humanos , Recién Nacido , Reproducibilidad de los Resultados , Fototerapia/métodos , Ictericia/terapiaAsunto(s)
Infecciones por Klebsiella , Osteomielitis , Humanos , Klebsiella pneumoniae , República de CoreaRESUMEN
BACKGROUND/AIM: It has been found that microRNAs (miRNA) affect rheumatoid arthritis (RA) pathophysiology. This study aimed to identify novel serum exosomal miRNAs related to RA disease activity in patients with an inadequate treatment response. PATIENTS AND METHODS: The sample population comprised clinical remission (CR) and non-clinical remission (non-CR) groups of RA patients. To identify potent miRNA markers for RA disease activity, miRNA array and qPCR were performed after patient serum exosomes preparation. RESULTS: Has-miR-1915-3p and has-miR-6511b-5p were significantly higher in the serum exosomes of the CR group. The level of serum C-reactive protein (CRP) was negatively correlated with has-miR-1915-3p level in serum exosomes. CONCLUSION: Has-miR-1915-3p may be a potential marker for Korean RA disease activity.
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Artritis Reumatoide , Exosomas , MicroARNs/sangre , Artritis Reumatoide/diagnóstico , Artritis Reumatoide/genética , Biomarcadores , Humanos , República de CoreaRESUMEN
PURPOSE: Rheumatoid arthritis (RA) is an autoimmune disease in which autoantibodies attack the synovial membrane, causing joint inflammation. Blood tests would offer a powerful, minimally invasive method for early diagnosis of RA. However, no reliable biomarkers for RA are presently available. The aim is to develop biomarkers for RA by multiple reaction monitoring (MRM)-based quantification of candidate biomarkers. EXPERIMENTAL DESIGN: Proteomics approaches are commonly used to identify and verify disease biomarkers. For discovery of biomarkers for RA, SWATH acquisition is performed and selected candidate biomarkers are validated by MRM. Target serum proteins are compared between patients with RA and healthy controls divided into three groups based on rheumatoid factor level. RESULTS: A total of 45 differentially expressed proteins are identified, as determined by SWATH acquisition. Of these, 13 proteins are selected as novel candidate biomarkers. A total of five proteins (transthyretin, gelsolin, angiotensinogen, lipopolysaccharide-binding protein, and protein S100-A9) are shown to have the potential to distinguish patients with RA from healthy controls. CONCLUSIONS AND CLINICAL RELEVANCE: These five proteins may improve the efficiency of diagnosis of RA. MRM can be used to easily diagnose RA by detecting five proteins simultaneously in a single sample with high sensitivity.
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Artritis Reumatoide/metabolismo , Proteómica , Anciano , Artritis Reumatoide/sangre , Biomarcadores/sangre , Estudios de Casos y Controles , Femenino , Perfilación de la Expresión Génica , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Analysis of the components of bloodstains found at crime scenes can provide important information for solving the crime. However, components of blood and bloodstains vary with volume and various other unpredictable factors. Therefore, it is necessary to specify the volume of the initial liquid blood droplet and standardize the analysis. In this study, internal standard metabolites that remained constant in a certain amount of bloodstain, long after deposition of the stain, were identified. Liquid chromatography-electrospray ionization-tandem mass spectrometry of the metabolites extracted from the bloodstain samples at various time points (0, 7, 14, 21, and 28 days) was performed. The coefficient of variation (CV) of the obtained molecular features was calculated for each criterion: time point, subject, and all data (time and subject, triplicate of each). Five molecular features with average CVs of less than or equal to 5% were selected as candidates. Partial least squares discriminant analysis and principal component analysis showed that the effect on the candidates was very low over time. The fold-change value of abundances was confirmed according to time. Stigmasterol exhibited the most stable pattern; l-methionine remained stable until day 14 and after day 21. This study was the first attempt to identify internal standard metabolites that were maintained at a constant level in a bloodstain for a sufficiently long time. Analysis of internal standard metabolites in bloodstains will facilitate determination of the initial blood volume from which the bloodstain was made. Moreover, this method will provide an approach for standardization of bloodstains to obtain absolute quantitative information of bloodstain components at crime scenes.
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Manchas de Sangre , Sangre/metabolismo , Metaboloma , Cromatografía Liquida , Análisis Discriminante , Femenino , Medicina Legal , Humanos , Análisis de los Mínimos Cuadrados , Masculino , Espectrometría de Masas en Tándem , Factores de Tiempo , Adulto JovenRESUMEN
Bloodstains are common evidence in crime scenes, containing significant information, including genetic information. Although efforts have been made to reliably determine the time of incident by analyzing the elapsed time of the bloodstain, there has been limited success. To identify candidate metabolites in bloodstains over time, we prepared bloodstain samples using filter paper and analyzed the metabolites by high-performance liquid chromatography-mass spectrometry (HPLC-MS)/MS over a 21-day period. Using Venn diagrams and by multivariate analysis, we selected 62 candidate molecular features. We found by partial least-squares discriminant analysis (PLS-DA) that the group can be classified with an accuracy of 75.0%, and the R2 and Q2 values were 0.7513 and 0.6998, respectively. Five metabolites were successfully identified based on candidate molecular features. The level of two metabolites, l-tryptophan and ergothioneine, decreased with time. The concentration of candidate metabolites that we propose reliably increased or decreased with time, thus, enabling the measurement of elapsed time of the bloodstain. This study is the first to identify markers used to analyze the elapsed time of bloodstains through metabolomics analysis.
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Ergotioneína/análisis , Metabolómica , Triptófano/análisis , Manchas de Sangre , Cromatografía Líquida de Alta Presión , Ergotioneína/metabolismo , Humanos , Análisis de los Mínimos Cuadrados , Espectrometría de Masas , Análisis Multivariante , Papel , Triptófano/metabolismoRESUMEN
BACKGROUND: We examined the feasibility of a full-length gene analysis for the drug resistance-related genes inhA, katG, rpoB, pncA, rpsL, embB, eis, and gyrA using ion semiconductor next-generation sequencing (NGS) and compared the results with those obtained from conventional phenotypic drug susceptibility testing (DST) in multidrug-resistant Mycobacterium tuberculosis (MDR-TB) isolates. METHODS: We extracted genomic DNA from 30 pure MDR-TB isolates with antibiotic susceptibility profiles confirmed by phenotypic DST for isoniazid (INH), rifampin (RIF), ethambutol (EMB), pyrazinamide (PZA), amikacin (AMK), kanamycin (KM), streptomycin (SM), and fluoroquinolones (FQs) including ofloxacin, moxifloxacin, and levofloxacin. Enriched ion spheres were loaded onto Ion PI Chip v3, with 30 samples on a chip per sequencing run, and Ion Torrent sequencing was conducted using the Ion AmpliSeq TB panel (Life Technologies, USA). RESULTS: The genotypic DST results revealed good agreement with the phenotypic DST results for EMB (Kappa 0.8), PZA (0.734), SM (0.769), and FQ (0.783). Agreements for INH, RIF, and AMK+KM were not estimated because all isolates were phenotypically resistant to INH and RIF, and all isolates were phenotypically and genotypically susceptible to AMK+KM. Moreover, 17 novel variants were identified: six (p.Gly169Ser, p.Ala256Thr, p.Ser383Pro, p.Gln439Arg, p.Tyr597Cys, p.Thr625Ala) in katG, one (p.Tyr113Phe) in inhA, five (p.Val170Phe, p.Thr400Ala, p.Met434Val, p.Glu812Gly, p.Phe971Leu) in rpoB, two (p.Tyr319Asp and p.His1002Arg) in embB, and three (p.Cys14Gly, p.Asp63Ala, p.Gly162Ser) in pncA. CONCLUSIONS: Ion semiconductor NGS could detect reported and novel amino acid changes in full coding regions of eight drug resistance-related genes. However, genotypic DST should be complemented and validated by phenotypic DSTs.
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Mycobacterium tuberculosis/genética , Análisis de Secuencia de ADN/métodos , Amicacina/farmacología , Antituberculosos/farmacología , Proteínas Bacterianas/genética , ADN Bacteriano/química , ADN Bacteriano/aislamiento & purificación , Farmacorresistencia Bacteriana Múltiple/genética , Genotipo , Humanos , Kanamicina/farmacología , Pruebas de Sensibilidad Microbiana , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/aislamiento & purificación , Fenotipo , Reacción en Cadena de la Polimerasa , Semiconductores , Análisis de Secuencia de ADN/instrumentación , Tuberculosis Resistente a Múltiples Medicamentos/diagnóstico , Tuberculosis Resistente a Múltiples Medicamentos/microbiologíaRESUMEN
BACKGROUND: Rheumatoid arthritis (RA) is an autoimmune disease that starts with inflammation of the synovial membrane. Studies have been conducted to develop methods for efficient diagnosis of RA and to identify the mechanisms underlying RA development. Blood samples can be useful for detecting disturbance of homeostasis in patients with RA. Nanoliquid chromatography-tandem mass spectrometry (LC-MS/MS) is an efficient proteomics approach to analyze blood sample and quantify serum proteins. METHODS: Serum samples of 18 healthy controls and 18 patients with RA were analyzed by LC-MS/MS. Selected candidate biomarkers were validated by enzyme-linked immunosorbent assay (ELISA) using sera from 43 healthy controls and 44 patients with RA. RESULTS: Thirty-eight proteins were significantly differentially expressed by more than 2-fold in healthy controls and patients with RA. Based on a literature survey, we selected six candidate RA biomarkers. ELISA was used to evaluate whether these proteins effectively allow distinguishing patients with RA from healthy controls and monitoring drug efficacy. SAA4, gelsolin, and vitamin D-binding protein were validated as potential biomarkers of RA for screening and drug efficacy monitoring of RA. CONCLUSIONS: We identified a panel of three biomarkers for RA which has potential for application in RA diagnosis and drug efficacy monitoring. Further, our findings will aid in understanding the pathogenesis of RA.
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Artritis Reumatoide/metabolismo , Biomarcadores/metabolismo , Anciano , Artritis Reumatoide/sangre , Biomarcadores/sangre , Estudios de Casos y Controles , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proteómica/métodosRESUMEN
Aceclofenac has been used widely as a potent analgesic and anti-inflammatory drug. Aceclofenac is converted to 4'-hydroxyaceclofenac and diclofenac via CYP2C9-mediated hydroxylation and hydrolysis, respectively. CYP2C9 also mediates the hydroxylation of diclofenac to yield 4'-hydroxydiclofenac and the hydrolysis of 4'-hydroxyaceclofenac to 4'-hydroxydiclofenac. We aimed to model the metabolism of aceclofenac in volunteers using a compartmental modeling approach. After an oral dose of 100 mg aceclofenac in volunteers, plasma concentrations of aceclofenac and its three metabolites were measured. The pharmacokinetics of aceclofenac and the sequential formation of its three metabolites were analyzed using ADAPT 5. The delay parameter shifted the plasma aceclofenac concentration-time profile to the right and provided a large improvement of fit. Two compartments were needed to fit the aceclofenac and 4'-hydroxyaceclofenac data, and one additional compartment was sufficient to describe the time courses of the generated plasma concentrations of diclofenac and 4'-hydroxydiclofenac. The metabolism rate constant for 4'-hydroxyaceclofenac was much greater than that for diclofenac. The generation rate constant of 4'-hydroxydiclofenac from diclofenac was greater than that of its generation from 4'-hydroxyaceclofenac. Our model fully describes the time course of plasma aceclofenac concentration as well as the formation and disposition of its three major metabolites in volunteers.
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Antiinflamatorios no Esteroideos/farmacocinética , Diclofenaco/análogos & derivados , Adulto , Antiinflamatorios no Esteroideos/sangre , Diclofenaco/sangre , Diclofenaco/farmacocinética , Voluntarios Sanos , Humanos , Masculino , Modelos Biológicos , Adulto JovenRESUMEN
It has been suggested that tumor cells secrete exosomes to modify the local microenvironment, which then promotes intercellular communication and metastasis. Although exosomes derived from cancer cells may contribute to the epithelial-mesenchymal transition (EMT) in untransformed cells, few studies have defined exosome cargo upon induction of EMT. In this study, we investigated the changes in exosomal cargo from the epithelial to mesenchymal cell phenotype by inducing EMT with transforming growth factor (TGF)-ß1 in A549 human lung adenocarcinoma cells. The protein content of the exosomes reflects the change in the cell phenotype. In addition, miR-23a was significantly enriched in the exosomes after mesenchymal transition. Following treatment of exosomes from mesenchymal cells via EMT induction with TGF-ß1 to the epithelial cell type, phenotypic changes in protein expression level and cell morphology were observed. Autologous treatment of exosomes enhanced the transcriptional activity and abundance of ß-catenin. Our results suggest that the exosomal protein and miRNA content reflects the physiological condition of its source and that exosomes induce phenotypic changes via autocrine signaling.
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Transición Epitelial-Mesenquimal/efectos de los fármacos , Exosomas/efectos de los fármacos , MicroARNs/genética , Factor de Crecimiento Transformador beta1/farmacología , beta Catenina/genética , Células A549 , Comunicación Autocrina , Transición Epitelial-Mesenquimal/genética , Exosomas/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , MicroARNs/metabolismo , Transducción de Señal , beta Catenina/metabolismoRESUMEN
BACKGROUND: Mutation of ABO glycosyltransferase (GT) can cause protein stability changes that can result in a weak ABO phenotype. To explain the Bw phenotype of a novel ABO*Bw allele, a protein stability of the mutant GT, which enhances the information of the three-dimensional (3D) structural analysis, was calculated. STUDY DESIGN AND METHODS: ABO serology and genotyping were performed on a neonate and her five family members. A 3D structural analysis of the wild-type GTB and enzymes with a variety of mutations at Residue 168, along with predicted protein stability changes (ΔΔG) and flow cytometric analysis of ABO antigen expression on HeLa cells transfected with plasmids containing R168Q, R168L, and R168P mutants was also performed. RESULTS: A novel ABO*Bw allele (c.503G>A, p.R168Q) was discovered. The structural analysis of 3D homology modeling predicted reduced protein stability of the mutant GTB, and the ΔΔG values, which inversely correlated with the mean relative fluorescence intensity of ABO antigen expression, quantitatively explained the reduced ABO antigen expression. CONCLUSIONS: The predicted protein stability change of a mutant GT enzyme might be a useful and convenient approach to objectively and quantitatively explain the reduced ABO antigen expression.
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Sistema del Grupo Sanguíneo ABO/genética , Glicosiltransferasas/genética , Mutación , Estabilidad de Enzimas , Genotipo , Glicosiltransferasas/química , Células HeLa , Humanos , Recién Nacido , FenotipoRESUMEN
Culture is the gold standard for diagnosis of tuberculosis, but it takes 6 to 8 weeks to confirm the result. This issue is complemented by the detection method using polymerase chain reaction, which is now widely used in a routine microbiology laboratory. In this study, we evaluated the performance of the Seegene Anyplex TB PCR to assess its diagnostic sensitivity and specificity, and compared its results with the Roche Cobas TaqMan MTB PCR, one of the most widely used assays in the world. Five university hospitals located in the Chungcheong area in South Korea participated in the study. A total of 1,167 respiratory specimens ordered for acid-fast bacilli staining and culture were collected for four months, analyzed via the Seegene Anyplex TB PCR, and its results were compared with the Roche Cobas TaqMan MTB PCR. For detection of Mycobacterium tuberculosis, the diagnostic sensitivity and specificity of the Anyplex TB PCR were 87.5% and 98.2% respectively, whereas those of the Cobas TaqMan were 92.0% and 98.0% respectively (p value > 0.05). For smear-positive specimens, the sensitivity of the Anyplex TB PCR was 95.2%, which was exactly the same as that of the Cobas TaqMan. For smear-negative specimens, the sensitivity of the Anyplex TB PCR was 69.2%, whereas that of the Cobas TaqMan TB PCR was 84.6%. For detection of MTB, the Seegene Anyplex TB PCR showed excellent diagnostic performance, and high sensitivity and specificity, which were comparable to the Roche Cobas TaqMan MTB PCR. In conclusion, the Anyplex TB PCR can be a useful diagnostic tool for the early detection of tuberculosis in clinical laboratories.
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Tipificación Molecular/métodos , Mycobacterium tuberculosis/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Tuberculosis Pulmonar/microbiología , Humanos , Mycobacterium tuberculosis/aislamiento & purificación , Sensibilidad y EspecificidadRESUMEN
In bloodstain pattern analysis, the blood droplet volume and surface impact velocity play an important role, and many related experimental studies have been carried out. If an appropriate computational fluid dynamics (CFD) model that could solve bloodstain patterns, especially spine formation bloodstain patterns, can be obtained, the blood droplet volume and impact speeds at various crime scenes can be predicted more accurately. For this purpose, Flow-3D software using the volume-of-fluid method was applied to analyze the behavior of human blood droplets during an impact event, especially focusing on the viscous effect on splashing, which forms the spine which can be used to predict the impact velocity. To obtain a non-Newtonian viscosity model of blood for a computational fluid dynamic analysis, the venous blood samples of 163 people were tested using a hemorheology instrument. Among the venous blood samples of 163 people, 37 samples for which all blood test results were in a normal range were selected for the non-Newtonian viscosity modeling. From the CFD analysis, it could be concluded that a non-Newtonian viscosity model is more appropriate than a constant viscosity model for predicting splashing that forms the spine. The gradient of the non-Newtonian model at a high shear rate has more of an effect on spine formation than that at a low shear rate. The lowest viscosity with a high velocity at the outer front of the radiating flow plays an important role in forming the splashing pattern.
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Manchas de Sangre , Biología Computacional , Simulación por Computador , Hemorreología , Humanos , Imagenología Tridimensional , Modelos Biológicos , Modelos Estadísticos , Programas InformáticosRESUMEN
OBJECTIVES: Dried blood spot (DBS) technology is a microsampling alternative to traditional plasma or serum sampling for pharmaco- or toxicokinetic evaluation. DBS technology has been applied to diagnostic screening in drug discovery, nonclinical, and clinical settings. We have developed an improved elution protocol involving boiling of blood spots dried on Whatman filter paper. METHODS: The purpose of this study was to compare the quality, purity, and quantity of DNA isolated from frozen blood samples and DBSs. We optimized a method for extraction and estimation of DNA from blood spots dried on filter paper (3-mm FTA card). A single DBS containing 40 µL blood was used. RESULTS: DNA was efficiently extracted in phosphate-buffered saline (PBS) or Tris-EDTA (TE) buffer by incubation at 37°C overnight. DNA was stable in DBSs that were stored at room temperature or frozen. The housekeeping genes GAPDH and beta-actin were used as positive standards for polymerase chain reaction (PCR) validation of general diagnostic screening. CONCLUSION: Our simple and convenient DBS storage and extraction methods are suitable for diagnostic screening by using very small volumes of blood collected on filter paper, and can be used in biobanks for blood sample storage.
RESUMEN
We have proposed a novel mobile healthcare platform, combining a pocket-sized colorimetric reader (13.5 × 6.5 × 2.5 cm(3)) and commercially available 10-parameter urinalysis paper strips (glucose, protein, glucose, bilirubin, urobilinogen, ketones, nitrite, pH, specific gravity, erythrocytes, and leukocytes), capable of sending data with a smart phone. The reader includes a novel colorimetric multi-detection module, which consists of three-chromatic light-emitting diodes, silicon photodiodes and a novel poly(methylmethacrylate) (PMMA) optical splitter. We employed data reading methods using conversions of the signal data (red, blue, and green) to the hue (H) color map or the Y model data, and used a curve-fitting method for the quantification. The reader is battery-powered, inexpensive, light-weight, and very speedy in analysis. And, it was applied to detection of a thousand of human urine samples and demonstrated reliable quantification of urinary glucose and protein. The features can be used by unskilled people on-site to transfer the analyzed data to experts off-site.
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Telemedicina/instrumentación , Urinálisis/instrumentación , Colorimetría/economía , Colorimetría/instrumentación , Diseño de Equipo , Humanos , Telemedicina/economía , Urinálisis/economíaRESUMEN
Approximately 10% of patients with chronic lymphocytic leukaemia (CLL) have a family history of the disease or a related lymphoproliferative disorder, yet the relationship of familial CLL to genomic abnormalities has not been characterized in detail. We therefore studied 75 CLL patients, half familial and half sporadic, using high-resolution array comparative genomic hybridization (CGH), in order to better define the relationship of genomic abnormalities to familial disease and other biological prognostic factors. Our results showed that the most common high-risk deletion in CLL, deletion 11q, was significantly associated with sporadic disease. Comparison of familial to sporadic disease additionally identified a copy number variant region near the centromere on 14q, proximal to IGH@, in which gains were associated both with familial CLL, and with mutated IGHV and homozygous deletion of 13q. Homozygous deletion of 13q was also found to be associated with mutated IGHV and low expression of ZAP-70, and a significantly longer time to first treatment compared to heterozygous deletion or lack of alteration. This study is the first high resolution effort to investigate and report somatic genetic differences between familial and sporadic CLL.
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Aberraciones Cromosómicas , Leucemia Linfocítica Crónica de Células B/genética , Adulto , Anciano , Deleción Cromosómica , Cromosomas Humanos Par 11/genética , Cromosomas Humanos Par 13/genética , Cromosomas Humanos Par 14/genética , Hibridación Genómica Comparativa/métodos , Femenino , Predisposición Genética a la Enfermedad , Humanos , Hibridación Fluorescente in Situ , Masculino , Persona de Mediana EdadRESUMEN
BACKGROUND: A primary inhibitor of plasminogen activators, SERPINE1 (serpin peptidase inhibitor 1, clade E, member 1, also known as plasminogen activator inhibitor type 1), is an important regulator in tumorigenesis and is highly expressed in many cancers. METHODS: Five tag single nucleotide polymorphisms (SNPs) and 1 insertion polymorphism within SERPINE1 were genotyped in 1101 unrelated Korean individuals (a case group of 612 patients with gastric cancer and a control group of 489 healthy individuals). Associations with susceptibility to diffuse-type gastric cancer (DGC) and intestinal-type gastric cancer were assessed using multivariate logistic regression analyses adjusted for age and sex. Allelic differences in the contribution to gene expression were measured using luciferase assays. RESULTS: SNP c.1162+162C>T (rs2227692) in intron 7 was associated with susceptibility to DGC but not with susceptibility to intestinal-type gastric cancer. The minor allele-carrying genotypes C/T and T/T had 1.6-fold greater odds of DGC than the C/C genotype (P = .00084). This SNP was linked to a repeat-number variation c.1162+604AAAG(11_17), a deletion (del) variation c.1162+664_1162+673del, and another SNP c.1162+859T>A (rs2070683) in intron 7 based on the sequencing of 5 patients and 5 controls. The risk haplotype of the 4 variations exhibited a 30% greater gene expression level than the nonrisk haplotype in luciferase reporter assays (P = .025). In contrast, DGC susceptibility was not associated with the c.-1969_-1968insG polymorphism (rs1799768) in the promoter, commonly known as 4G/5G, in which the minor 5G allele is less active in transcription than the major 4G allele. CONCLUSIONS: An association between SERPINE1 and DGC susceptibility was observed with 4 correlated polymorphisms in intron 7 rather than the 4G/5G polymorphism in the promoter, although all polymorphisms affected gene expression.
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Predisposición Genética a la Enfermedad , Inhibidor 1 de Activador Plasminogénico/genética , Polimorfismo Genético , Neoplasias Gástricas/genética , Adulto , Anciano , Anciano de 80 o más Años , Progresión de la Enfermedad , Femenino , Expresión Génica , Humanos , Intrones , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Neoplasias Gástricas/patologíaRESUMEN
BACKGROUND: The diagnosis of septic arthritis (SA) relies on synovial analysis and conventional culture. But, these methods lack of sensitivity and culture is time consuming to establish a definite diagnosis. This study evaluated a new multiplex PCR assay which entailed screening PCR for Gram typing and identification PCR for species identification using two primer mixes. METHODS: A total of 80 synovial fluid samples from patients with suspected SA were collected. Culture, multiplex PCR, and 16S rRNA gene PCR were performed. RESULTS: The analytical sensitivity of multiplex PCR assay was 10(1) CFU/ml for each type of bacteria. There was no cross-reactivity with common bacterial pathogens. Bacteria were detected in 20, 25, and 26 of 80 samples for culture, multiplex PCR, and 16S rRNA gene PCR, respectively. Nineteen (95%) of 20 culture-positive samples and 6 (10%) of 60 culture-negative samples were positive for the multiplex PCR. Five of six samples which were positive only from multiplex PCR were also positive in 16S rRNA gene PCR. The multiplex PCR showed 2 false-negative in 27 true-positive samples but no false-positive. The sensitivity and specificity of the multiplex PCR were 92.6 and 100%, and the agreement with culture and 16S rRNA gene PCR were 91.3 and 96.3%, respectively. The time to detection for multiplex PCR was a maximum of 6 hr. CONCLUSION: This multiplex PCR assay offers high sensitivity and improved detection speed relative to culture. The appropriate combination of this new multiplex PCR assay with culture may contribute to the accurate and rapid diagnosis of SA.
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Artritis Infecciosa/diagnóstico , Infecciones Bacterianas/diagnóstico , Tipificación Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Artritis Infecciosa/microbiología , Infecciones Bacterianas/microbiología , Candida/genética , Candida/aislamiento & purificación , Reacciones Falso Negativas , Bacterias Gramnegativas/genética , Bacterias Gramnegativas/aislamiento & purificación , Bacterias Grampositivas/genética , Bacterias Grampositivas/aislamiento & purificación , Humanos , ARN Ribosómico 16S/genética , Sensibilidad y Especificidad , Líquido Sinovial/microbiologíaRESUMEN
The B3 phenotype is the most common B subtype in Korea. The B305 allele (425 T>C, M142T) was first reported in 2 Chinese individuals; however, it has not yet been reported in the Koreans, and the impact of the M142T mutation on the expression of the B3 phenotype has also not been studied. To resolve an ABO discrepancy between a group O neonate and her group O father and A(1)B(3) mother, blood samples from these individuals and other family members were referred to our laboratory for ABO gene analysis. The B305 allele was discovered in the neonate (B305/O01), her mother (A102/ B305), and her maternal aunt (B305/O02), while her father was typed as O01/O02. Transient transfection experiments were performed in HeLa cells using the B305 allele synthesized by site-directed mutagenesis; flow cytometric analysis revealed that this transfect expressed 35.5% of the total B antigen produced by the B101 allele transfect. For comparison, Bx01 allele transfects were also created, and they expressed 11.4% of the total B antigen expressed on the surface of B101 transfects. These experiments demonstrate that the M142T (425 T>C) mutation is responsible for the B subtype phenotype produced by the B305 allele.