RESUMO
Glucosinolates (GSLs) are defensive secondary metabolites produced by Brassicaceae species in response to abiotic and biotic stresses. The biosynthesis of GSL compounds and the expression of GSL-related genes are highly modulated by endogenous signals (i.e., circadian clocks) and environmental cues, such as temperature, light, and pathogens. However, the detailed mechanism by which light signaling influences GSL metabolism remains poorly understood. In this study, we found that a light-signaling factor, ELONGATED HYPOCOTYL 5 (HY5), was involved in the regulation of GSL content under light conditions in Arabidopsis (Arabidopsis thaliana). In hy5-215 mutants, the transcript levels of GSL pathway genes were substantially upregulated compared with those in wild-type plants. The content of GSL compounds was also substantially increased in hy5-215 mutants, whereas 35S::HY5-GFP/hy5-215 transgenic lines exhibited comparable levels of GSL-related transcripts and GSL content to those in WT plants. HY5 physically interacts with HISTONE DEACETYLASE9 (HDA9) and binds to the proximal promoter region of MYB29 and IMD1 to suppress aliphatic GSL biosynthetic processes. These results demonstrate that HY5 suppresses GSL accumulation during the daytime, thus properly modulating GSL content daily in Arabidopsis plants.
RESUMO
As sessile organisms, plants encounter dynamic and challenging environments daily, including abiotic/biotic stresses. The regulation of carbon and nitrogen allocations for the synthesis of plant proteins, carbohydrates, and lipids is fundamental for plant growth and adaption to its surroundings. Light, one of the essential environmental signals, exerts a substantial impact on plant metabolism and resource partitioning (i.e., starch). However, it is not fully understood how light signaling affects carbohydrate production and allocation in plant growth and development. An orphan gene unique to Arabidopsis thaliana, named QUA-QUINE STARCH (QQS) is involved in the metabolic processes for partitioning of carbon and nitrogen among proteins and carbohydrates, thus influencing leaf, seed composition, and plant defense in Arabidopsis. In this study, we show that PHYTOCHROME-INTERACTING bHLH TRANSCRIPTION FACTORS (PIFs), including PIF4, are required to suppress QQS during the period at dawn, thus preventing overconsumption of starch reserves. QQS expression is significantly de-repressed in pif4 and pifQ, while repressed by overexpression of PIF4, suggesting that PIF4 and its close homologs (PIF1, PIF3, and PIF5) act as negative regulators of QQS expression. In addition, we show that the evening complex, including ELF3 is required for active expression of QQS, thus playing a positive role in starch catabolism during night-time. Furthermore, QQS is epigenetically suppressed by DNA methylation machinery, whereas histone H3 K4 methyltransferases (e.g., ATX1, ATX2, and ATXR7) and H3 acetyltransferases (e.g., HAC1 and HAC5) are involved in the expression of QQS. This study demonstrates that PIF light signaling factors help plants utilize optimal amounts of starch during the night and prevent overconsumption of starch before its biosynthesis during the upcoming day.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Fitocromo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fitocromo/metabolismo , Amido/metabolismo , Carbono/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Nitrogênio/metabolismo , Regulação da Expressão Gênica de Plantas , Luz , Arseniato Redutases/genética , Arseniato Redutases/metabolismoRESUMO
Floral transition is accelerated by exposure to long-term cold like winter in plants, which is called as vernalization. Acceleration of floral transition by vernalization is observed in a diversity of biennial and perennial plants including Brassicaceae family plants. Scientific efforts to understand molecular mechanism underlying vernalization-mediated floral transition have been intensively focused in model plant Arabidopsis thaliana. To get a better understanding on floral transition by vernalization in radish (Raphanus sativus L.), we investigated transcriptomic changes taking place during vernalization in radish. Thousands of genes were differentially regulated along time course of vernalization compared to non-vernalization (NV) sample. Twelve major clusters of DEGs were identified based on distinctive expression profiles during vernalization. Radish FLC homologs were shown to exert an inhibition of floral transition when transformed into Arabidopsis plants. In addition, DNA region containing RY motifs located within a Raphanus sativus FLC homolog, RsFLC1 was found to be required for repression of RsFLC1 by vernalization. Transgenic plants harboring disrupted RY motifs were impaired in the enrichment of H3K27me3 on RsFLC1 chromatin, thus resulting in the delayed flowering in Arabidopsis. Taken together, we report transcriptomic profiles of radish during vernalization and demonstrate the requirement of RY motif for vernalization-mediated repression of RsFLC homologs in radish (Raphanus sativus L.).
Assuntos
Arabidopsis , Brassicaceae , Raphanus , Raphanus/genética , Arabidopsis/genética , Vernalização , CromatinaRESUMO
BACKGROUND AND AIMS: This study aimed to validate Helicobacter pylori serological and pepsinogen (PG) assays for detecting infection and gastric neoplasm. METHODS: Individuals who underwent serum Chorus H. pylori and HBI PG assays were included from May to September 2023. The GastroPanel test was performed using the same blood sample. HBI assay findings were interpreted with the ABC method using the criteria of corpus atrophy (PG I ≤ 70 ng/mL & I/II ≤3) and advanced corpus atrophy (PG I ≤ 30 ng/mL & I/II ≤2). RESULTS: A total of 144 H. pylori-infected and 184 non-infected Koreans were analyzed. The Chorus test (sensitivity 97.2%, specificity 89.1%) showed higher area under the curve (0.993 vs. 0.972, p = 0.003) than the GastroPanel test (sensitivity 95.8%, specificity 86.4%). Using the GastroSoft application, the incidence of gastric neoplasms was highest in the corpus atrophy group (50%), followed by the low acid-output (25.8%), H. pylori infection (11.6%), and antral atrophy (9.1%) groups. There were no gastric neoplasms in the normal and high acid output groups. Using the ABC method, the incidence of gastric neoplasms was highest in the corpus atrophy groups (23.8% in Groups C and D), followed by Group B (12.3%) and Group A (2.4%). Corpus atrophy interpreted with the GastroSoft showed poor agreement (k = 0.225) with corpus atrophy interpreted with the ABC method, whereas it showed excellent agreement (k = 0.854) with advanced corpus atrophy. CONCLUSIONS: Although the Chorus test was more accurate than the GastroPanel test, both assays discriminated high-risk individuals by detecting atrophy or infection. There were no gastric neoplasms in the normal or high acid-output groups (GastroSoft application), and gastric neoplasm incidence was lowest in Group A (ABC method). Corpus atrophy determined by GastroSoft application is more consistent with advanced corpus atrophy determined by the ABC method than is corpus atrophy.
Assuntos
Infecções por Helicobacter , Helicobacter pylori , Neoplasias Gástricas , Humanos , Pepsinogênio A , Estudos Prospectivos , Infecções por Helicobacter/diagnóstico , AtrofiaRESUMO
OBJECTIVES: White blood cell (WBC)-related flags are essential for detecting abnormal cells including blasts in automated hematology analyzers (AHAs). Cell population data (CPD) may characterize each WBC population, and customized CPD rules can be also useful for detecting blasts. We evaluated the performance of WBC-related flags, customized CPD rules, and their combination for detecting blasts on the Beckman Coulter DxH 900 AHA (DxH 900, Beckman Coulter, Miami, Florida, USA). METHODS: In a total of 239 samples from patients with hematologic diseases, complete blood count on DxH 900 and manual slide review (MSR) were conducted. The sensitivity, specificity, and efficiency of the five WBC-related flags, nine customized CPD rules, and their combination were evaluated for detecting blasts, in comparison with MSR. RESULTS: Blasts were detected by MSR in 40 out of 239 (16.7â¯%) samples. The combination of flags and CPD rules showed the highest sensitivity compared with each of flags and CPD rules for detecting blasts (97.5 vs. 72.5â¯% vs. 92.5â¯%). Compared with any flag, the combination of flags and CPD rules significantly reduced false-negative samples from 11 to one for detecting blasts (27.5 vs. 2.5â¯%, p=0.002). CONCLUSIONS: This is the first study that evaluated the performance of both flags and CPD rules on DxH 900. The customized CPD rules as well as the combination of flags and CPD rules outperformed WBC-related flags for detecting blasts on DxH 900. The customized CPD rules can play a complementary role for improving the capability of blast detection on DxH 900.
Assuntos
Doenças Hematológicas , Hematologia , Humanos , Contagem de Células Sanguíneas , Doenças Hematológicas/diagnóstico , Leucócitos , Contagem de LeucócitosRESUMO
Methylglyoxal (MG) is a toxic by-product of the glycolysis pathway in most living organisms and was previously shown to inhibit seed germination. MG is detoxified by glyoxalase I and II family proteins in plants. MG is abundantly produced during early embryogenesis in Arabidopsis seeds. However, the mechanism that alleviates the toxic effect of MG in maturing seeds is poorly understood. In this study, by T-DNA mutant population screening, we found that mutations in a glyoxalase I gene (named GERMINATION-IMPAIRED GLYOXALASE 1, GIG1) led to significantly impaired germination compared with wild-type seeds. Transformation of full-length GIG1 cDNA under the constitutively active cauliflower mosaic virus 35S promoter in the gig1 background completely recovered the seed germination phenotype. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analyses revealed that GIG1 is uniquely expressed in seeds and is upregulated by abscisic acid (ABA) and downregulated by gibberellic acid (GA) during seed germination. An ABA signaling component, ABI3, directly activated GIG1 in maturing seeds. In addition, PHYTOCHROME INTERACTING FACTOR 1 (PIF1) also plays cooperatively with ABI3 in the regulation of GIG1 expression in the early stage of imbibed seeds. Furthermore, GIG1 expression is stably silenced by epigenetic repressors such as polycomb repressor complexes. Altogether, our results indicate that light and ABA signaling cooperate to enhance seed germination by the upregulation of GIG1 to detoxify MG in maturing seeds.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Lactoilglutationa Liase , Fitocromo , Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Regulação da Expressão Gênica de Plantas , Germinação , Lactoilglutationa Liase/genética , Lactoilglutationa Liase/metabolismo , Fitocromo/metabolismo , Aldeído Pirúvico/metabolismo , Sementes/metabolismo , Fatores de Transcrição/metabolismoRESUMO
BACKGROUND AND OBJECTIVES: VISION Max (Ortho Clinical Diagnostics, Raritan, NJ) measures anti-A/B isoagglutinin titres using automated column agglutination technology (CAT). We compared tube test (TT) and CAT of VISION Max comprehensively, including failure mode and effect analysis (FMEA), turnaround time (TAT) and cost, and suggested modified CAT (MCAT). MATERIALS AND METHODS: For 100 samples (each 25 for blood type A, B and O with anti-A and anti-B), anti-A/B isoagglutinin titres were measured by TT and CAT (1:2-1:1024 dilution), as well as by MCAT (with agglutination at 1:32 dilution, then perform additional testing from 1:64 to 1:1024). We assessed the agreement and correlation between TT and CAT and compared FMEA (risk priority number [RPN] score), TAT (h:min:sec) and cost (US dollar, US $) among TT, CAT and MCAT. RESULTS: TT and CAT showed overall substantial agreement (k = 0.73) and high correlation (ρ ≥ 0.75) except blood type O with anti-A (ρ = 0.68). Compared with TT, CAT showed lower RPN scores in FMEA and similar TAT and cost (FMEA, 33,700 vs. 184,300; TAT, 15:23:00 vs. 14:26:40; cost, 1377.4 vs. 1312.4, respectively). Regarding FMEA, TAT and cost, MCAT was superior to CAT or TT (43,810; 13:28:00; 899.2, respectively). CONCLUSION: This is the first multidimensional analysis on VISION Max CAT for measuring anti-A/B isoagglutinin titres. The results of anti-A/B isoagglutinin titres by CAT were comparable with those of TT. MCAT would be a safe, time-saving and cost-effective alternative to TT and CAT in high-volume blood bank laboratories.
Assuntos
Sistema ABO de Grupos Sanguíneos , Hemaglutininas , Aglutinação , Anticorpos , TecnologiaRESUMO
Strontium (Sr) has become an increasing global threat for both environment and human health due to its radioactive isotope, Sr-90 which can be found in the nuclear-contaminated soils and water. Although excessive Sr has been known to be toxic to plant growth and development, the molecular mechanisms underlying plant response to Sr stress, especially on the transcription level, remains largely unknown. To date, there is no published genome-wide transcriptome data available for the plant responses to Sr toxicity. Therefore, we aimed to gain insight on the molecular events occurring in plants in Sr toxicity condition by comparing the genome-wide gene expression profiles between control and Sr-treated plants using RNA-seq analysis. A total of 842 differentially expressed genes (DEGs) were identified in response to Sr stress compared to the control. Based on the analysis of DEGs using Gene Ontology (GO), DEGs were significantly enriched in the GO terms of response to salicylic acid (SA), response to jasmonic acid (JA), and defense response to bacterium. In addition, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis indicated that DEGs were mainly involved in metabolic processes including phenylpropanoid biosynthesis and alpha-linolenic acid metabolism, which is known as a precursor of JA biosynthesis. Furthermore, MapMan analysis revealed that a number of genes related to the biotic stress such as pathogenesis-related protein (PR) genes were highly up-regulated under Sr stress. Taken together, this study revealed that JA biosynthesis and/or signaling might be associated with plant response to Sr stress, and play important roles to maintain proper growth and development under Sr stress.
Assuntos
Oxilipinas , Estrôncio , Ciclopentanos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Humanos , Oxilipinas/metabolismo , Estrôncio/metabolismo , TranscriptomaRESUMO
OBJECTIVES: Vision Pro (West Medica, Perchtoldsdorf, Austria) is a recently developed digital morphology analyzer. We evaluated the performance of Vision Pro on white blood cell (WBC) differentials. METHODS: In a total of 200 peripheral blood smear samples (100 normal and 100 abnormal samples), WBC preclassification and reclassification by Vision Pro were evaluated and compared with manual WBC count, according to the Clinical and Laboratory Standards Institute guidelines (H20-A2). RESULTS: The overall sensitivity was high for normal WBCs and nRBCs (80.1-98.0%). The overall specificity and overall efficiency were high for all cell classes (98.1-100.0% and 97.7-99.9%, respectively). The absolute values of mean differences between Vision Pro and manual count ranged from 0.01 to 1.31. In leukopenic samples, those values ranged from 0.09 to 2.01. For normal WBCs, Vision Pro preclassification and manual count showed moderate or high correlations (r=0.52-0.88) except for basophils (r=0.34); after reclassification, the correlation between Vision Pro and manual count was improved (r=0.36-0.90). CONCLUSIONS: This is the first study that evaluated the performance of Vision Pro on WBC differentials. Vision Pro showed reliable analytical performance on WBC differentials with improvement after reclassification. Vision Pro could help improve laboratory workflow.
Assuntos
Leucócitos , Projetos de Pesquisa , Contagem de Células Sanguíneas , Contagem de Leucócitos , Padrões de ReferênciaRESUMO
We evaluated the performance of new high-throughput digital lateral flow immunoassays (LFIAs) detecting influenza antigens and compared them with those of the widely used digital LFIA and the rapid nucleic acid amplification test (NAAT). We tested 199 clinical nasopharyngeal (nasal) swab samples using three LFIA tests (BD Veritor Plus, STANDARD F Influenza A/B FIA, and ichroma TRIAS) and the rapid NAAT (ID NOW Influenza A & B2). Agreements and clinical performances (sensitivity and specificity) were evaluated based on the results of reverse transcriptase-polymerase chain reaction (RT-PCR) and verification panel. The agreement of each test with RT-PCR was moderate to almost perfect. The sensitivity of ID NOW was significantly higher than that of LFIAs (P = .0005, .0044, and .0026 for influenza A and P = .0044, .0026, and .0044 for influenza B, respectively). The specificities were not significantly different between the four tests (P > .05). However, the reference panel suggests that ichroma TRIAS test is more sensitive than the other two LFIA tests. All three LFIA assays performed similarly with no false positives against influenza A. For influenza B, ichroma TRIAS had 2 of 166 false positives whereas there were no false positives for the other two LFIA tests. Influenza antigen digital LFIAs have advantages in terms of the workflow when simultaneous tests are required. Rapid NAAT has higher sensitivity, while new antigen LFIAs are efficient and high-throughput. It is recommended that users select appropriate methods and algorithms according to the number of specimens and laboratory conditions in each clinical laboratory.
Assuntos
Antígenos Virais/análise , Imunoensaio/métodos , Vírus da Influenza A/isolamento & purificação , Vírus da Influenza B/isolamento & purificação , Influenza Humana/diagnóstico , Nasofaringe/virologia , Reações Falso-Positivas , Humanos , Vírus da Influenza A/imunologia , Vírus da Influenza B/imunologia , Influenza Humana/virologia , Técnicas de Amplificação de Ácido Nucleico , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Manejo de EspécimesRESUMO
Soil moisture-precipitation feedbacks in a large ensemble of global climate model simulations are evaluated. A set of three metrics are used to assess the sensitivity of afternoon rainfall occurrence to morning soil moisture in terms of their spatial, temporal, and heterogeneity characteristics. Positive (negative) spatial feedback indicates that the afternoon rainfall occurs more frequently over wetter (drier) land surface than its surroundings. Positive (negative) temporal feedback indicates preference over temporally wetter (drier) conditions, and positive (negative) heterogeneity feedback indicates preference over more spatially heterogeneous (homogeneous) soil moisture conditions. We confirm previous results highlighting a dominantly positive spatial feedback in the models as opposed to observations. On average, models tend to agree better with observations for temporal and heterogeneity feedback characteristics, although intermodel variability is largest for these metrics. The collective influence of the three feedbacks suggests that they may lead to more localized precipitation persistence in models than in observations.
RESUMO
OBJECTIVES: The aim of this study was to investigate the diagnostic accuracy of rheumatoid factor (RF) isotype for the detection of primary Sjogren's syndrome (pSS) and evaluate the clinical and serological associations of immunoglobulin (Ig) A RF in patients with pSS. MATERIALS AND METHODS: RF levels were measured in 77 and 37 patients with pSS and idiopathic sicca symptoms, respectively, using ELISA and analysed with respect to clinical and laboratory disease characteristics. Receiver operating characteristic curves were used to determine and compare the diagnostic accuracy of IgA RF with other diagnostic tests. RESULTS: Serum levels of IgA RF were significantly higher in patients with pSS than in those with idiopathic sicca symptoms. IgA RF showed sensitivity, specificity, positive, and negative predictive values of 83.1, 78.4, 88.9, and 69.0%, respectively, for pSS diagnosis. IgA RF was associated with xerostomia, severe sialoscintigraphic grade, low unstimulated salivary flow rate (USFR), antinuclear antibody, high IgG and IgM/G RF levels, and low C3 levels in patients with pSS. IgA RF titres had positive correlations with sialoscintigraphic grade and IgG and IgG/M RF levels and had negative correlations with USFR and C3 levels. CONCLUSION: Our findings confirmed the potential of IgA RF to distinguish pSS from idiopathic sicca symptoms. The presence of IgA RF in patients with pSS was associated with significantly worse exocrine function and active serologic profile. No association between IgA RF and extra-glandular manifestations was noted. CLINICAL RELEVANCE: IgA RF should be the predictive and diagnostic marker in patients with pSS.
Assuntos
Imunoglobulina A/sangue , Fator Reumatoide/sangue , Síndrome de Sjogren/diagnóstico , Idoso , Anticorpos Antinucleares/sangue , Feminino , Humanos , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Síndrome de Sjogren/sangueRESUMO
PURPOSE: This study aimed to evaluate the clinical significance of toxin positivity and toxin gene load, and the relation between them in the broad spectrum of Clostridium difficile infection (CDI) including colonization, significant diarrhea, and severe disease. METHODS: We included 2671 fecal samples submitted for CDI diagnosis and 180 samples from healthy individuals. The clinical spectrum was categorized as category I (toxigenic C. difficile positive without clinical CDI criteria), category II (mild CDI), and category III (severe CDI). Clinical parameters were compared based on toxin EIA and tcdB C t values. C t values of tcdB PCR for predicting toxin EIA positivity were assessed using receiver-operating characteristic (ROC) curves. RESULTS: The median C t values of tcdB PCR and toxin positivity were not significantly correlated with clinical spectrum of CDI (27.5, 28.2, and 26.1 for tcdB C t and 55.0, 56.6, and 60.9% for toxin EIA positivity in category I, II, and III, respectively, P > 0.05). There were significant differences in the tcdB C t values between toxin EIA-positive and -negative groups (P < 0.001). Optimal cutoff for the tcdB C t value for estimating toxin EIA positivity was 26.3 with 79.3% sensitivity and 83.6% specificity with good area under the curves (AUC, 0.848). CONCLUSIONS: The C t values successfully predicted toxin EIA positivity and could be used as a surrogate for toxin EIA positivity in the diagnostic algorithm and routine analysis. Further studies are needed to validate the clinical significance of tcdB PCR C t value in toxigenic C. difficile colonization and infection.
Assuntos
Proteínas de Bactérias/genética , Toxinas Bacterianas/genética , Clostridioides difficile/fisiologia , Infecções por Clostridium/microbiologia , Diarreia/microbiologia , Carga Genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Bactérias/metabolismo , Toxinas Bacterianas/metabolismo , Clostridioides difficile/genética , Fezes/microbiologia , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Anti-cyclic citrullinated peptide antibodies (anti-CCPs) are important diagnostic markers for rheumatoid arthritis (RA). We evaluated the analytical and clinical performance of the QUANTA Flash CCP3 (INOVA Diagnostics, USA), a fully automated third-generation anti-CCP assay, in comparison with three second-generation anti-CCP (CCP2) assays. A total of 300 sera (67 from RA patients, 64 from other rheumatic diseases, 43 from osteoarthritis [OA], and 126 from other conditions) were tested with QUANTA Flash CCP3, Kallestad Anti-CCP II (Bio-Rad, USA), Elecsys Anti-CCP (Roche Diagnostics GmbH, Germany), and ARCHITECT Anti-CCP (Abbott Diagnostics, USA). Within-run and total imprecision (% coefficient of variation) of the QUANTA Flash CCP3 were <6%, and its linearity was acceptable over the claimed range (4.0-2,749.7 chemiluminescent units). The frequency of anti-CCP was similar between QUANTA Flash CCP3 and the other CCP2 assays in the RA (67.2% vs. 62.7-70.1%), other rheumatic diseases (7.8% vs. 6.3%), and OA (2.3% vs. 0-2.3%) groups. The concordance rate between QUANTA Flash CCP3 and the other assays ranged from 96.3% to 97.7% (kappa from 0.87 to 0.92). For the diagnosis of RA, the sensitivity/specificity was 67.2%/95.7%, 62.7%/98.3%, 70.2%/96.6%, and 67.2%/97.9%, and the areas under the receiver operating characteristic curves were 0.851, 0.791, 0.853, and 0.867 for QUANTA Flash CCP3, Kallestad, Elecsys, and ARCHITECT assays, respectively. The performance of the QUANTA Flash CCP3 was satisfactory and comparable to that of the three CCP2 assays. This fully automated assay would be a practical and reasonable option in clinical laboratories.
Assuntos
Anticorpos Antiproteína Citrulinada/sangue , Artrite Reumatoide/diagnóstico , Automação Laboratorial/normas , Imunoensaio/normas , Osteoartrite/diagnóstico , Adulto , Idoso , Artrite Reumatoide/sangue , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Biomarcadores/sangue , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite/sangue , Osteoartrite/imunologia , Osteoartrite/patologia , Curva ROCRESUMO
Although launched in 2015, little is known about the accuracy of QuantiFERON-TB Gold-Plus (QFT-Plus) for diagnosis of latent M. tuberculosis infection (LTBI). Unlike its predecessor, QFT-Plus utilizes two antigen tubes to elicit an immune response from CD4+ and CD8+ T lymphocytes. We conducted a cross-sectional study in low-risk health care workers (HCWs) at a single U.S. center to compare QFT-Plus to QuantiFERON-TB Gold in-tube (QFT). A total of 989 HCWs were tested with both QFT and QFT-Plus. Risk factors for LTBI were obtained from a questionnaire. QFT-Plus was considered positive if either antigen tube 1 (TB1) or TB2 tested positive, per the manufacturer's recommendations, or if both TB1 and TB2 tested positive, using a conservative definition. Results were compared using Cohen's kappa and linear regression, respectively. Agreement of QFT with QFT-Plus was high, at 95.6% (95% confidence interval [CI], 94.3 to 96.9; kappa, 0.57). The majority of discordant results between QFT and QFT-Plus TB1 (84.8%) and QFT and QFT-Plus TB2 (88.6%) fell within the range of 0.2 to 0.7 IU/ml. The positivity rate in 626 HCWs with no identifiable risk factors and no self-reported history of positive LTBI tests was 2.1% (CI, 1.0 to 3.2) and 3.0% (CI, 1.7 to 4.3) with QFT and QFT-Plus, respectively. A conservative definition of a QFT-Plus-positive result yielded a positivity rate of 1.0% (CI, 0.2 to 1.7; P value of 0.0002 versus QFT-Plus and 0.07 versus QFT). On follow-up testing, of 11 HCWs with discordant QFT-Plus results, 90.9% (10/11) had a negative QFT result. The QFT-Plus assay showed a high degree of agreement with QFT in U.S. HCWs. A conservative interpretation of QFT-Plus eliminated nearly all nonreproducible positive results in low-risk HCWs. Larger studies are needed to validate the latter finding and to more clearly define conditions under which a conservative interpretation can be used to minimize nonreproducible positive results in low-risk populations.
Assuntos
Testes Diagnósticos de Rotina/métodos , Pessoal de Saúde , Testes de Liberação de Interferon-gama/métodos , Tuberculose Latente/diagnóstico , Adulto , Estudos Transversais , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Inquéritos e Questionários , Estados UnidosRESUMO
BACKGROUND: The Sysmex DI-60 system (DI-60, Sysmex, Kobe, Japan) is a new automated digital cell imaging analyzer. We explored the performance of DI-60 in comparison with Sysmex XN analyzer (XN, Sysmex) and manual count. METHODS: In a total of 276 samples (176 abnormal and 100 normal samples), white blood cell (WBC) differentials, red blood cell (RBC) classification and platelet (PLT) estimation by DI-60 were compared with the results by XN and/or manual count. RBC morphology between pre-classification and verification was compared according to the ICSH grading criteria. The manual count was performed according to the Clinical and Laboratory Standards Institute guidelines (H20-A2). RESULTS: The overall concordance between DI-60 and manual count for WBCs was 86.0%. The agreement between DI-60 pre-classification and verification was excellent (weighted κ=0.963) for WBC five-part differentials. The correlation with manual count was very strong for neutrophils (r=0.955), lymphocytes (r=0.871), immature granulocytes (r=0.820), and blasts (r=0.879). RBC grading showed notable differences between DI-60 and manual counting on the basis of the ICSH grading criteria. Platelet count by DI-60 highly correlated with that by XN (r=0.945). However, DI-60 underestimated platelet counts in samples with marked thrombocytosis. CONCLUSIONS: The performance of DI-60 for WBC differential, RBC classification, and platelet estimation seems to be acceptable even in abnormal samples with improvement after verification. DI-60 would help optimize the workflow in hematology laboratory with reduced manual workload.
Assuntos
Automação , Testes Hematológicos , Contagem de Células Sanguíneas , Eritrócitos/citologia , Testes Hematológicos/instrumentação , Humanos , Leucócitos/citologia , Linfócitos/citologiaRESUMO
BACKGROUND: The correlation between allergic disease and Helicobacter pylori infection is still controversial in endemic areas. The aim of this study was to determine whether H. pylori infection is related to allergic disease and/or immunoglobulin E (IgE) hypersensitivity in Korean adults. MATERIALS AND METHODS: Consecutive Korean adults who visited our center for a routine checkup were enrolled. All subjects completed a questionnaire that was designed to ascertain their medical history pertaining to physician-diagnosed allergic disease, allergy treatments, and H. pylori eradication therapy. Blood was sampled for serum anti-H. pylori IgG antibody. IgE hypersensitivity was measured using a commercially available ImmunoCAP(®) Phadiatop (Phadia AB, Uppsala, Sweden). RESULTS: Of the 3376 Korean adults who were enrolled, 62 did not answer to the questionnaires adequately and were thus excluded. The proportion of noninfected subjects (p < .001) and the prevalence of IgE-related allergic disease (p < .001) were both highest among those aged <40 years, while the prevalence of non-IgE-related allergic disease was highest among those aged ≥70 years (p < .001). Logistic regression analysis revealed that being younger than 40 years was significantly related to the absence of H. pylori infection (OR = 2.507, 95% CI = 1.621-3.878, p < .001). CONCLUSIONS: The statuses of H. pylori infection, IgE hypersensitivity, and allergic diseases differ with age group, there being a higher prevalence of IgE-related allergic disease and a lower H. pylori infection rate among young adults. The hygiene hypothesis might explain these findings in young Koreans, due to the rapid development and improvements in sanitation in Korea.
Assuntos
Infecções por Helicobacter/complicações , Hipersensibilidade/epidemiologia , Imunoglobulina E/sangue , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Povo Asiático , Estudos Transversais , Feminino , Humanos , Coreia (Geográfico)/epidemiologia , Masculino , Pessoa de Meia-Idade , Inquéritos e Questionários , Adulto JovemRESUMO
BACKGROUND: Recently introduced hematology analyzer, the Sysmex XN modular system (Sysmex, Kobe, Japan), has newly adopted a florescent channel to detect platelets and immature platelet fraction (IPF). This study aimed to establish new reference intervals for %-IPF and absolute number of IPF (A-IPF) on Sysmex XN. Platelet counts, %-IPF, and A-IPF were also compared between Sysmex XN and XE-2100 systems (Sysmex). METHODS: Except outliers, blood samples from 2104 healthy individuals and 140 umbilical cord blood were analyzed using both Sysmex XN and XE-2100. The results of two systems were compared using Bland-Altman plot. The reference intervals for %-IPF and A-IPF were defined using non-parametric percentile methods according to the Clinical and Laboratory Standard Institute guideline (C28-A3). RESULTS: The platelet counts, %-IPF, and A-IPF showed non-parametric distributions. The mean difference between Sysmex XN and XE-2100 in healthy individuals revealed a positive bias in platelets (+8.0×109/L), %-IPF (+1.2%), and A-IPF (+3.0×109/L). The reference intervals for %-IPF and A-IPF on Sysmex XN were: 1.0%-7.3% and 2.49-15.64×109/L in healthy individuals; and 1.0%-4.4% and 2.94-12.82×109/L in umbilical cord blood. CONCLUSIONS: This large-scale study demonstrates a clear difference of platelet counts and IPF between Sysmex XN and XE-2100. The new reference intervals for IPF on Sysmex XN would provide fundamental data for clinical practice and future research.