Fast and accurate pathogen detection in aquatic environments is challenging in many biomedical studies and microbial diagnostic applications. In this study, we developed a real-time, continuous, and ...non-destructive single cell detection method using target specific aptamer-conjugated fluorescent nanoparticles (A-FNPs) and an optofluidic particle-sensor platform. A-FNPs selectively bound to the surfaces of target bacteria (Escherichia coli) and labeled them with high affinity and selectivity so that target bacteria can be countable particles in an optofluidic particle-sensor. A-FNP-labeled target bacterial complexes were detected by the optofluidic particle-sensing system, which provides rapid and continuous single-cell detection. A-FNPs selectively bound to E. coli with a dissociation constant of 0.83 nM, but did not bind Enterobacter aerogenes or Citrobacter freundii strains, which lacked affinity for the aptamer used. We demonstrated that our optofluidic device achieves a detection throughput of ~100 particles per second with high accuracy (~85%) in detecting single bacterial cells conjugated with A-FNPs. This approach can be immediately extended to the real-time, high-throughput detection of other microorganisms such as viruses that are selectively conjugated with A-FNPs. Collectively, these data suggest that optofluidic systems are widely applicable for the fast and continuous detection of microbial cells.
Clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease (Cas9) can be used for the specific disruption of a target gene to permanently suppress the expression ...of the protein encoded by the target gene. Efficient delivery of the system to an intracellular target site should be achieved to utilize the tremendous potential of the genome-editing tool in biomedical applications such as the knock-out of disease-related genes and the correction of defect genes. Here, we devise polymeric CRISPR/Cas9 system based on poly-ribonucleoprotein (RNP) nanoparticles consisting of polymeric sgRNA, siRNA, and Cas9 endonuclease in order to improve the delivery efficiency. When delivered by cationic lipids, the RNP nanoparticles built with chimeric poly-sgRNA/siRNA sequences generate multiple sgRNA-Cas9 RNP complexes upon the Dicer-mediated digestion of the siRNA parts, leading to more efficient disruption of the target gene in cells and animal models, compared with the monomeric sgRNA-Cas9 RNP complex.
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Watson for oncology (WFO) is a cognitive computing system providing decision support. We evaluated the concordance rates between the treatment options determined by WFO and those determined by a ...multidisciplinary team (MDT).
We reviewed the medical charts of patients diagnosed with colorectal cancer who visited the MDT at a single tertiary medical center from November 2016 to April 2017. WFO classified the treatment options for specific patients into three categories: 'Recommended', 'For consideration', and 'Not recommended'. Concordance rates between the WFO- and MDT-determined chemotherapy options, and the factors that potentially influence the concordance rate, were analyzed.
Sixty-nine patients with colorectal cancer met with the MDT from Nov. 2016 to Feb. 2017. The mean age of the patients was 62 years (range: 34-86 years), and more patients were male (47/69) than female. Of the 69 patients, 51 (73.9%) were diagnosed with colon cancer, of whom 46.4% received the same regimen recommendation from WFO ('Recommended') as they did from the MDT. After inclusion of the 'For consideration' category from WFO, the concordance rate increased to 87.0%. The concordance rate between MDT and NCCN guidelines was 97.1%, and that between the WFO and NCCN guidelines was 88.4%. The concordance rates between WFO and MDT were significantly lower in patients with stage II, IIIC, or IV disease (P<0.001), and the colorectal cancer stage was the only statistically significant factor discriminating between WFO and MDT.
The concordance rate between chemotherapy regimens for colorectal cancer determined by MDT versus WFO recommendations was 46.4%. After including the 'For consideration' category from WFO, the concordance rate increased to 88.4%. Further modification and improvement of the WFO prioritizing algorithm used to recommend treatment may increase the usefulness of WFO in the clinic.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
: This study evaluated the effectiveness of short fully covered self-expanding metal stents (FCSEMS) with an anti-migration design in treating benign biliary strictures (BBS) not related to living ...donor liver transplantation (LDLT).
: A retrospective analysis was conducted on 75 patients who underwent FCSEMS insertion for BBS management. Stents were initially kept for 3 months and exchanged every 3 months until stricture resolution. Adverse events and stricture recurrence after FCSEMS removal were assessed during follow-up.
: The study outcomes were technical success, stenosis resolution, and treatment failure. Technical success was 100%, with stricture resolution in 99% of patients. The mean onset time of BBS post-surgery was 4.4 years, with an average stent indwelling period of 5.5 months. Stricture recurrence occurred in 20% of patients, mostly approximately 18.8 months after stent removal. Early cholangitis and stent migration were noted in 3% and 4% of patients, respectively.
: This study concludes that short FCSEMS demonstrate high efficacy in the treatment of non-LDLT-related BBS, with a low incidence of interventions and complications. Although this is a single-center, retrospective study with a limited sample size, the findings provide preliminary evidence supporting the use of short FCSEMS as a primary treatment modality for BBS. To substantiate these findings, further research involving multicenter studies is recommended to provide additional validation and a broader perspective.
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The aryl hydrocarbon receptor (AHR) is a ligand-activated signaling molecule which is involved in diverse biological functions ranging from cancer metastasis to immune regulation. ...This receptor forms a cytoplasmic complex with Hsp90, p23, and XAP2. We have previously reported that down-regulation of p23 triggers degradation of the AHR protein, uncovering a potentially dynamic event which controls the cellular AHR levels without ligand treatment. Here we investigate the underlying mechanisms for this p23 effect using wild-type HeLa and the p23 knockdown HeLa cells. Reduction of the Hsp90 and XAP2 contents, however, did not affect the AHR protein levels, implying that this p23 effect on AHR is more than just alteration of the cytoplasmic complex dynamics. Association of p23 with Hsp90 is not important for the modulation of the AHR levels since exogenous expression of p23 mutants with modest Hsp90-binding affinity effectively restored the AHR message and protein levels. The protein folding property of p23 which resides at the terminal 50-amino acid region is not involved for this p23 effect. Results from our interaction study using the affinity purified thioredoxin fusion proteins and GST fusion proteins showed that p23 directly interacts with AHR and the interaction surface lies within AHR amino acid 1–216 and p23 amino acid 1–110. Down-regulation of the p23 protein content promotes the ubiquitination of AHR, indicating that p23 protects AHR from the ubiquitin-meditated protein degradation.
Abstract
The healthcare industry is in dire need of rapid microbial identification techniques for treating microbial infections. Microbial infections are a major healthcare issue worldwide, as these ...widespread diseases often develop into deadly symptoms. While studies have shown that an early appropriate antibiotic treatment significantly reduces the mortality of an infection, this effective treatment is difficult to practice. The main obstacle to early appropriate antibiotic treatments is the long turnaround time of the routine microbial identification, which includes time-consuming sample growth. Here, we propose a microscopy-based framework that identifies the pathogen from single to few cells. Our framework obtains and exploits the morphology of the limited sample by incorporating three-dimensional quantitative phase imaging and an artificial neural network. We demonstrate the identification of 19 bacterial species that cause bloodstream infections, achieving an accuracy of 82.5% from an individual bacterial cell or cluster. This performance, comparable to that of the gold standard mass spectroscopy under a sufficient amount of sample, underpins the effectiveness of our framework in clinical applications. Furthermore, our accuracy increases with multiple measurements, reaching 99.9% with seven different measurements of cells or clusters. We believe that our framework can serve as a beneficial advisory tool for clinicians during the initial treatment of infections.
Self-assembled polymeric nanocapsules (NCs) that incorporate dendrimer porphyrin (DP) in the shells and superparamagnetic iron oxide nanoparticles (SPIONs) in the cores are fabricated to create a ...theragnostic platform for the application in photodynamic therapy (PDT) and magnetic resonance imaging (MRI). SPIONs-embedded polystyrene NPs (SPIONs@PS) are used as a template to build up multilayered NCs. The formation of PAH/DP multilayer on the SPIONs@PS is monitored by zeta-pential and fluorescence emission measurement, because the porphyrin unit in the core of DP has strong red fluorescence emission. NCs have strong enough magnetic property (>20 emu/g) for MRI application with typical superparamagnetic behavior, where the linear correlation of R2 and Fe concentration at diluted conditions led to corresponding T2 relaxivity coefficient (r2) value of 93.5 mM–1 s–1. Cell viability study upon light irradiation reveals that NCs can successfully work in photosensitizer formulation for PDT.
Information about the natural history of nontuberculous mycobacterial pulmonary disease (NTM-PD) is limited. The purpose of this study was to evaluate the long-term natural history of non-cavitary ...nodular bronchiectatic NTM-PD and the factors associated with treatment initiation and the frequency of spontaneous sputum culture conversion after diagnosis of NTM-PD.
We evaluated 1,021 patients with newly diagnosed non-cavitary nodular bronchiectatic NTM-PD caused by Mycobacterium avium complex or M. abscessus between 2003 and 2013.
Of 1,021 patients, 562 (55%) initiated antibiotic treatment and 459 (45%) did not. Young age (adjusted hazard ratio aHR = 0.99; 95% confidence interval CI = 0.98–0.99), low body mass index (aHR = 0.96; 95% CI = 0.93–0.99), previous history of tuberculosis (aHR = 1.23; 95% CI = 1.01–1.50), respiratory complaints such as cough (aHR = 1.36; 95% CI = 1.05–1.75) and sputum production (aHR = 1.47; 95% CI = 1.13–1.91), and high number of involved lobes on high-resolution computed tomography (aHR = 1.22; 95% CI = 1.14–1.31) were associated with treatment initiation. Of 459 patients who did not initiate treatment, 157 (34%) showed spontaneous sputum culture conversion. None of the clinical factors was associated with spontaneous conversion. After spontaneous culture conversion, 26 of 157 (17%) showed redeveloped NTM-PD caused by a species different from the original species.
The natural history of non-cavitary nodular bronchiectatic NTM-PD is variable. After diagnosis, the decision to initiate antibiotic therapy should be individualized based on consideration of the risk factors for disease progression. However, for patients who do not start antibiotic therapy, continuous and lifetime follow-up is recommended to manage underlying bronchiectasis and the possibility of late progression of NTM-PD.
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Point-of-care (POC) molecular diagnostics for clinical microbiology and virology has primarily focused on the detection of a single pathogen. More recently, it has transitioned into a comprehensive ...syndromic approach that employs multiplex capabilities, including the simultaneous detection of two or more pathogens. Multiplex POC tests provide higher accuracy to for actionable decisionmaking in critical care, which leads to pathogen-specific treatment and standardized usages of antibiotics that help prevent unnecessary processes. In addition, these tests can be simple enough to operate at the primary care level and in remote settings where there is no laboratory infrastructure. This review focuses on state-of-the-art multiplexed molecular point-of-care tests (POCT) for infectious diseases and efforts to overcome their limitations, especially related to inadequate throughput for the identification of syndromic diseases. We also discuss promising and imperative clinical POC approaches, as well as the possible hurdles of their practical applications as front-line diagnostic tests.
Background Blood pressure (BP) variability has been reported to be associated with hypertensive target organ damage and cardiovascular events. However, the exact mechanism linking BP variability and ...organ damage is uncertain. This study was designed to investigate the association between BP variability and inflammatory marker in hypertensive patients. Methods and Results Fifty-two hypertensive patients (28 men, 55.9±1.5 years) completed 24-h ambulatory BP monitoring. Inflammatory markers were evaluated by measuring plasma levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay and high sensitive C-reactive protein (hs-CRP) by particle-enhanced light-scattering immunoassay. BP variability was obtained by calculating within-subject standard deviation (SD) and coefficient of variation of BP. Subjects were grouped into tertiles according to IL-6, TNF-α, and hs-CRP levels. A significant association between ambulatory BP and TNF-α level was identified (P for trend =0.011). In contrast, no association was observed between BP and IL-6 level; however, BP variability index was linked to IL-6 level (P for trend =0.046). The association between inflammatory marker and pattern of diurnal variation was investigated. The hs-CRP concentration was significantly higher in the riser group compared with the dipper group. However, IL-6 and TNF-α levels did not differ among the different diurnal variation groups. Correlation analysis showed varying associations between IL-6 and TNF-α. TNF-α level correlated with the BP index; however, IL-6 level correlated with the BP variability index. Multiple linear regression models revealed that the SD of daytime systolic BP (β=0.065, p=0.001) and age (β=0.024, p=0.016) were all positively and significantly related to IL-6. In contrast, only daytime diastolic BP (β=0.029, p=0.002) was independently related to TNF-α. Conclusion Inflammatory markers are associated with BP variability in hypertensive patients. This finding implies that inflammation may be a mediator for the link between BP variability and target organ damage. (Circ J 2008; 72: 293 - 298)
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