scholarly journals A dextran-based warming method for preparing leukocyte-rich plasma and its clinical application for endotoxin assay

BioTechniques ◽  
2020 ◽  
Vol 68 (6) ◽  
pp. 300-304
Author(s):  
Gaku Takahashi ◽  
Katsuya Inada ◽  
Koutaro Sato ◽  
Yoshihiro Inoue

We devised a method using dextran for obtaining leukocyte-rich plasma (LRP) to measure endotoxin in blood. In order to find the optimal temperature for obtaining LRP, the measurement results were examined using samples prepared at 37 and 0°C. Sample separation time of LRP was significantly shorter at 37°C than at 0°C. Endotoxin measurement values showed a strong correlation between the two groups but many of the LRPs made at 37°C had measurements above those of the LRPs prepared at 0°C. The diagnostic accuracy for gram-negative bacterial infection was superior for LRP prepared at 37°C, with sensitivity and specificity of 96.8 and 100%, respectively.

2021 ◽  
pp. 1-11
Author(s):  
Bing-Jie Xiang ◽  
Min Jiang ◽  
Ming-Jun Sun ◽  
Cong Dai

<b><i>Objective:</i></b> Fecal calprotectin (FC) is a promising marker for assessment of inflammatory bowel disease (IBD) activity. However, the utility of FC for predicting mucosal healing (MH) of IBD patients has yet to be clearly demonstrated. The objective of our study was to perform a meta-analysis evaluating the diagnostic accuracy of FC in predicting MH of IBD patients. <b><i>Methods:</i></b> We systematically searched the databases for studies from inception to April 2020 that evaluated MH in IBD. The methodological quality of each study was assessed according to the Quality Assessment of Diagnostic Accuracy Studies checklist. The extracted data were pooled using a summary receiver operating characteristic curve model. Random-effects model was used to summarize the diagnostic odds ratio, sensitivity, specificity, positive likelihood ratio, and negative likelihood ratio. <b><i>Results:</i></b> Sixteen studies comprising 1,682 ulcerative colitis (UC) patients and 4 studies comprising 221 Crohn’s disease (CD) patients were included. The best performance of FC for predicting MH in UC was at cut-off range of 60–75 μg/g with area under the curve (AUC) of 0.88 and pooled sensitivity and specificity of 0.87 and 0.79, respectively. The pooled sensitivity and specificity values of cutoff range 180–250 μg/g for predicting MH in CD were 0.67 and 0.76, respectively. The AUC of 0.79 also revealed improved discrimination for identifying MH in CD with FC concentration. <b><i>Conclusion:</i></b> Our meta-analysis has found that FC is a simple, reliable noninvasive marker for predicting MH in IBD patients. FC cutoff range 60–75 μg/g appears to have the best overall accuracy in UC patients.


2018 ◽  
Vol 100-B (12) ◽  
pp. 1542-1550 ◽  
Author(s):  
J. van den Kieboom ◽  
P. Bosch ◽  
J. D. J. Plate ◽  
F. F. A. IJpma ◽  
R. Kuehl ◽  
...  

Aims To assess the diagnostic value of C-reactive protein (CRP), leucocyte count (LC), and erythrocyte sedimentation rate (ESR) in late fracture-related infection (FRI). Materials and Methods PubMed, Embase, and Cochrane databases were searched focusing on the diagnostic value of CRP, LC, and ESR in late FRI. Sensitivity and specificity combinations were extracted for each marker. Average estimates were obtained using bivariate mixed effects models. Results A total of 8284 articles were identified but only six were suitable for inclusion. Sensitivity of CRP ranged from 60.0% to 100.0% and specificity from 34.3% to 85.7% in all publications considered. Five articles were pooled for meta-analysis, showing a sensitivity and specificity of 77.0% and 67.9%, respectively. For LC, this was 22.9% to 72.6%, and 73.5% to 85.7%, respectively, in five articles. Four articles were pooled for meta-analysis, resulting in a 51.7% sensitivity and 67.1% specificity. For ESR, sensitivity and specificity ranged from 37.1% to 100.0% and 59.0% to 85.0%, respectively, in five articles. Three articles were pooled in meta-analysis, showing a 45.1% sensitivity and 79.3% specificity. Four articles analyzed the value of combined inflammatory markers, reporting an increased diagnostic accuracy. These results could not be pooled due to heterogeneity. Conclusion The serum inflammatory markers CRP, LC, and ESR are insufficiently accurate to diagnose late FRI, but they may be used as a suggestive sign in its diagnosis.


2020 ◽  
Author(s):  
Jia-Jin Chen ◽  
Chih-Hsiang Chang ◽  
Yen Ta Huang ◽  
George Kuo

Abstract Background: The use of the furosemide stress test (FST) as an acute kidney injury (AKI) severity marker has been described in several trials. However, the diagnostic performance of the FST in predicting AKI progression has not yet been fully discussed. Methods: In accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, we searched the PubMed, Embase, Cochrane databases up to March, 2020. The diagnostic performance of the FST (in terms of sensitivity, specificity, number of events, true positive, false positive) was extracted and evaluated. Results: We identified eleven trials that enrolled a total of 1366 patients, including 517 patients and 1017 patients for whom the outcomes in terms of AKI stage progression and renal replacement therapy (RRT), respectively, were reported. The pooled sensitivity and specificity results of the FST for AKI progression prediction were 0.81 (95% CI: 0.74 - 0.87) and 0.88 (95% CI: 0.82- 0.92), respectively. The pooled positive likelihood ratio (LR) was 5.45 (95% CI: 3.96-7.50), the pooled negative LR was 0.26 (95% CI: 0.19-0.36), and the pooled diagnostic odds ratio (DOR) was 29.69 (95% CI: 17.00-51.85). The summary receiver operating characteristics (SROC) with pooled diagnostic accuracy was 0.88. The diagnostic performance of the FST in predicting AKI progression was not affected by different AKI criteria or underlying chronic kidney disease. The pooled sensitivity and specificity results of the FST for RRT prediction were 0.84 (95% CI: 0.72-0.91) and 0.77 (95% CI: 0.64-0.87), respectively. The pooled positive LR and pooled negative LR were 3.16 (95% CI: 2.06-4.86) and 0.25 (95% CI: 0.14-0.44), respectively. The pooled diagnostic odds ratio (DOR) was 13.59 (95% CI: 5.74-32.17) and SROC with pooled diagnostic accuracy was 0.86. The diagnostic performance of FST for RRT prediction is better in stage 1-2 AKI comparing to stage 3 AKI (relative DOR: 5.75, 95% CI: 2.51-13.33) Conclusion: The FST is a simple tool for the identification of AKI populations at high risk of AKI progression and the need for RRT and the diagnostic performance of FST in RRT prediction is better in early AKI population.


2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Qiushuang Sheng ◽  
Runbao Du ◽  
Cunhui Ma ◽  
Yonglin Zhou ◽  
Xue Shen ◽  
...  

Abstract Background The wide spread of plasmid-mediated colistin resistance by mobile colistin resistance (MCR) in Enterobacteriaceae severely limits the clinical application of colistin as a last-line drug against bacterial infection. The identification of colistin potentiator from natural plants or their compound preparation as antibiotic adjuncts is a new promising strategy to meet this challenge. Methods Herein, the synergistic activity, as well as the potential mechanism, of Pingwei pill plus antibiotics against MCR-positive Gram-negative pathogens was examined using checkerboard assay, time-killing curves, combined disk test, western blot assay, and microscope analysis. Additionally, the Salmonella sp. HYM2 infection models of mouse and chick were employed to examine the in vivo efficacy of Pingwei pill in combination with colistin against bacteria infection. Finally, network pharmacology and molecular docking assay were used to predicate other actions of Pingwei pill for Salmonella infection. Results Our results revealed that Pingwei Pill synergistically potentiated the antibacterial activity of colistin against MCR-1-positive bacteria by accelerating the damage and permeability of the bacterial outer membrane with an FIC (Fractional Inhibitory Concentration) index less than 0.5. The treatment of Pingwei Pill neither inhibited bacterial growth nor affected MCR production. Notably, Pingwei Pill in combination with colistin significantly prolonged the median survival in mouse and chick models of infection using the Salmonella sp. strain HYM2, decreased bacteria burden and organ index of infected animal, alleviated pathological damage of cecum, which suggest that Pingwei Pill recovered the therapeutic performance of colistin for MCR-1- positive Salmonella infection in mice and the naturally infected host chick. Pharmacological network topological analysis, molecular docking, bacterial adhesion, and invasion pathway verification assays were performed to identify the other molecular mechanisms of Pingwei Pill as a colistin potentiator against Gram-negative bacteria infection. Conclusion Taken together, NMPA (National Medical Products Administration)-approved Pingwei Pill is a promising adjuvant with colistin for MCR-positive bacterial infection with a shortened R&D (research and development) cycle and affordable R&D cost and risk.


2018 ◽  
Author(s):  
Mahmoud M. Ashawesh ◽  
Robert Markus ◽  
Christopher N. Penfold ◽  
Kim R. Hardie

AbstractBacterial infection of humans, animals and plants relies heavily on secreted proteases that degrade host defences or activate bacterial toxins. The largest family of proteins secreted by Gram-negative pathogenic bacteria, the Autotransporters (ATs), includes key proteolytic virulence factors. There remains uncertainty about the mechanistic steps of the pathway ATs share to exit bacteria, and how it is energetically driven. This study set out to shed light on the AT secretion pathway with the ultimate aim of uncovering novel antimicrobial targets that would be unlikely to trigger the development of resistance mechanisms in bacteria. To do this, two AT virulence factors with distinct proteolytic functions, EspC (secreted from EnteropathogenicEscherichia coli) and AaaA (tethered to the extracellular surface ofPseudomonas aeruginosa) were chosen. EspC and AaaA were fluorescently labelled using two separate methods to establish the localization patterns of ATs as they are secreted from a bacterial cell. Super resolution microscopy revealed that localization of ATs occurs via a helical route along the bacterial cytoskeleton. In addition to requiring the conserved C-terminal β-barrel translocator domain of the AT, we present the first evidence that secretion is dependent on a dynamic interaction with a structure reliant upon the actin homologue MreB and the Sec translocon. These findings provide a step forward in the mechanistic understanding of the secretion of this widely distributed family of proteins that have pivotal roles in bacterial pathogenesis and conserved structural properties that could serve as novel broad-range antimicrobial targets.SignificanceSecreted bacterial proteases facilitate the infection of human, animal and plant hosts by degrading host defences or activating bacterial toxins. The autotransporter family is the largest family of proteins secreted from Gram-negative bacteria, and includes proteolytic virulence factors crucial to bacterial infection. Precisely how autotransporters migrate from the inside to the outside of the cell, and how this movement is energetically driven is a mystery. We demonstrate a spiral pathway of autotransporter secretion, presenting evidence that it involves a dynamic interaction with the actin homologue MreB that comprises the bacterial cytoskeleton. Our findings open the way to unravelling the mechanism of autotransporter secretion and offer the possibility to identify novel antimicrobial targets unlikely to trigger the development of antimicrobial resistance.


2020 ◽  
Vol 183 (2) ◽  
pp. 191-201
Author(s):  
Charlotte Michaela Fries ◽  
Yoon Ju Bae ◽  
Nada Rayes ◽  
Benjamin Sandner ◽  
Berend Isermann ◽  
...  

Objective Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) has become state of the art for the quantitative analysis of steroid hormones. Although method comparisons show that aldosterone measurement using LC-MS/MS yields considerably lower levels than immunoassays (IAs), method-specific cutoff values for primary aldosteronism (PA) are largely missing. Objective of this study was to analyze the diagnostic accuracy of proposed LC-MS/MS-specific cutoff values for the saline infusion test (SIT). Design and methods From 2016 to 2019, 104 consecutive patients suspected of PA underwent the SIT and captopril challenge test in the tertiary medical center at the University Hospital of Leipzig, Germany. Patients with positive case confirmation underwent adrenal imaging and adrenal venous sampling for subtype classification. Results Overall, proposed assay-specific PACLC-MS/MS cutoff values for the SIT achieved higher diagnostic accuracy than established PACIA values with a sensitivity and specificity of 87.5% (95% CI: 71.0–96.5) and 97% (95% CI: 89.6–99.6) for a cutoff of 120 pmol/L and 93.8% (95% CI: 79.2–99.2) and 92.5% (95% CI: 83.4–97.5) for a cutoff of 94 pmol/L. The most accurate post-SIT PACLC-MS/MS cutoff value in this study was 83 pmol/L, yielding a sensitivity and specificity of 96.9% (95% CI: 83.8–99.9) and 92.5% (95% CI: 83.4–97.5), respectively. Conclusions The present data confirm the need for the implication of lower method-specific aldosterone cutoff values for the diagnosis of PA with LC-MS/MS based aldosterone measurement.


Author(s):  
Firoozeh Ahmadi ◽  
Farnaz Akhbari ◽  
Fatemeh Niknejad ◽  
Hadieh Haghighi ◽  
Zahra Ghahremani ◽  
...  

ABSTRACT Introduction Two of the most frequent procedures performed on infertile women are two-dimensional ultrasound (2DUS) and three-dimensional ultrasound (3DUS). Hysteroscopy is considered as the gold standard for evaluation of acquired endometrial lesions in infertile women; however, 3DUS is used as a noninvasive, less expensive, and reliable assessment method for evaluation of the intrauterine lesions in infertile women. We aimed to compare the diagnostic efficiency between 3DUS and hysteroscopy in the detection of lesions (polyps, submucous leiomyoma, and synechiae) in infertile women. Materials and methods In this prospective observational study, infertile women (n = 155) with indication of hysteroscopy were scheduled to undergo 3DUS prior to hysteroscopy from September 2010 to 2011. Women with suspected congenital uterine anomalies were excluded. The sensitivity and specificity values of 3DUS were compared with those of hysteroscopy. Hysteroscopy was used as the gold standard for diagnosis of intrauterine lesions in infertile women. Results Of the 155 women, 50 were found to have an intracavitary abnormality, 36 had polyps, 12 had myomas, and 7 had synechiae on hysteroscopic findings. Examination with 3DUS in the diagnosis of intrauterine lesions reached an accuracy of 94%, and 92.15 and 96.9% of sensitivity and specificity respectively. Positive predictive value (PPV) was 83.9%, and a negative predictive value (NPV) was 91.3% (LR+ = 10.75, LR+ = 0.065). Conclusion According to our results, 3DUS has a reliable diagnostic accuracy for intrauterine lesions, and it may limit unnecessary hysteroscopy in patients with normal results. How to cite this article Ahmadi F, Haghighi H, Ghahremani Z, Niknejad F, Akhbari F, Ramezanali F, Chehrazi M. Diagnostic Accuracy of Three-dimensional Ultrasonography in Detection of Endometrial Lesions compared with Hysteroscopy in Infertile Women. Donald School J Ultrasound Obstet Gynecol 2016;10(4):393-397.


PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0253525
Author(s):  
Ashutosh Nath Aggarwal ◽  
Ritesh Agarwal ◽  
Sahajal Dhooria ◽  
Kuruswamy Thurai Prasad ◽  
Inderpaul Singh Sehgal ◽  
...  

Objective We compared diagnostic accuracy of pleural fluid adenosine deaminase (ADA) and interferon-gamma (IFN-γ) in diagnosing tuberculous pleural effusion (TPE) through systematic review and comparative meta-analysis. Methods We queried PubMed and Embase databases to identify studies providing paired data for sensitivity and specificity of both pleural fluid ADA and IFN-γ for diagnosing TPE. We used hierarchical summary receiver operating characteristic (HSROC) plots and HSROC meta-regression to model individual and comparative diagnostic performance of the two tests. Results We retrieved 376 citations and included 45 datasets from 44 publications (4974 patients) in our review. Summary estimates for sensitivity and specificity for ADA were 0.88 (95% CI 0.85–0.91) and 0.91 (95% CI 0.89–0.92), while for IFN-γ they were 0.91 (95% CI 0.89–0.94) and 0.96 (95% CI 0.94–0.97), respectively. HSROC plots showed consistently greater diagnostic accuracy for IFN-γ over ADA across the entire range of observations. HSROC meta-regression using test-type as covariate yielded a relative diagnostic odds ratio of 2.22 (95% CI 1.68–2.94) in favour of IFN-γ, along with better summary sensitivity and specificity figures. No prespecified subgroup variable significantly influenced the summary diagnostic accuracy estimates. Conclusion Pleural fluid IFN-γ estimation has better diagnostic accuracy than ADA estimation for diagnosis of TPE.


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