Quantification Of Activity Research Articles

Preschoolers’ Moderate-to-Vigorous Physical Activity Measured by a Tri-Axial Accelerometer: Compliance with International Guidelines and Different Cut-Points

Background/Objectives: This study aimed to investigate the effect of the most frequently used accelerometer CoPs on the quantification of active preschoolers by weekday; and to analyze children’s physical activity (PA) quantification using a vertical axis and vector magnitude (VM). Methods: A cross-sectional sample of 134 children (70 males) aged 3–5 years was studied. Height, body weight, and BMI were assessed. A tri-axial accelerometer was used for seven consecutive days of MVPA and sedentary behavior (SB). Data were analyzed using the three most used CoPs for active preschooler classification (Johansson, Butte, and Pate). A general linear model with repeated measures examined differences in PA and SB, and the agreements of all CoPs were analyzed using the Kappa index. Results: The CoPs adopted by Pate had the highest percentage of children classified as active for the weekdays (73.9%) and weekend (85.6%). The Johansson CoP classified all children as inactive. Furthermore, the prevalence of active boys was significantly higher than their female counterparts based on the Pate and Butte CoPs for the week and weekends. Conclusions: The lowest prevalence rates of active children were observed at the weekend based on all accelerometer CoPs, especially among girls. The choice of cut-points significantly affects the times calculated for different movement intensities.

Multimodal and quantitative analysis of the epileptogenic zone network in the pre-surgical evaluation of drug-resistant focal epilepsy

Surgical resection for epilepsy often fails due to incomplete Epileptogenic Zone Network (EZN) localization from scalp electroencephalography (EEG), stereo-EEG (SEEG), and Magnetic Resonance Imaging (MRI). Subjective interpretation based on interictal, or ictal recordings limits conventional EZN localization. This study employs multimodal analysis using high-density-EEG (HDEEG), Magnetoencephalography (MEG), functional-MRI (fMRI), and SEEG to overcome these limitations in a patient with drug-resistant MRI-negative focal epilepsy. A 17-year-old with drug-resistant epilepsy underwent evaluation. HDEEG, MEG, fMRI, and SEEG were used, with a novel HDEEG-cap facilitating simultaneous EEG-MEG and EEG-fMRI recordings. Electrical and magnetic source imaging were performed, and fMRI data were analysed for homogenous regions. SEEG analysis involved spike detection, spike timing analysis, ictal fast activity quantification, and Granger-based connectivity analysis. Non-invasive sessions revealed consistent interictal source imaging results identifying the EZN in the right anterior cingulate cortex. EEG-fMRI highlighted broader activation in the right cingulate cortex. SEEG analysis localized spikes and fast activity in the right anterior and posterior cingulate gyri. Multi-modal analysis suggested the EZN in the right frontal lobe, primarily involving the anterior and mid-cingulate cortices. Multi-modal non-invasive analyses can optimise SEEG implantation and surgical decision-making. Invasive analyses corroborated non-invasive findings, emphasising the importance of individual-case quantitative analysis across modalities in complex epilepsy cases.

Population Pharmacokinetic Quantification of CYP2D6 Activity in Codeine Metabolism in Ambulatory Surgical Patients for Model-Informed Precision Dosing.

Codeine metabolism in humans is complex due to the involvement of multiple cytochrome P450 (CYP) enzymes, and has a strong genetic underpinning, which determines the levels of relevant CYP450 enzyme expression in vivo. Polymorphic CYP2D6 metabolises codeine to morphine via O-demethylation, while a strong correlation between CYP2D6 phenotype and opioidergic adverse effects of codeine is well documented. The aim of this study was to quantify the effect of CYP2D6 genotype on the biotransformation of codeine. We conducted a prospective clinical trial with 1000 patients, during which ambulatory patients were administered 60 mg of codeine preoperatively and the association between CYP2D6 activity and morphine exposure across various CYP2D6 genotypes was quantified using a population pharmacokinetic model. Plasma concentration data for codeine and its primary metabolites were obtained from 997 patients and CYP2D6 genotype was screened for study subjects, and respective sums of activity scores assigned for each CYP2D6 allele were used as covariates in model development. Our final model predicts the disposition of codeine and the formation of morphine, codeine-6-glucuronide and morphine-3-glucuronide adequately while accounting for variability in morphine exposure on the basis of CYP2D6 genotype. In agreement with previous results, patients with decreased function alleles (CYP2D6*10 and *41) showed varying levels of decrease in CYP2D6 activity that were inconsistent with increasing activity scores. Model simulations demonstrate that morphine concentrations in ultrarapid CYP2D6 metabolisers reach systemic concentrations that can potentially cause respiratory depression (over 9.1 ng/mL), and have 218% higher exposure (19 versus 8.7 µg · h/L, p < 0.001) to morphine than normal metabolisers. Similarly, poor and intermediate metabolisers had significantly reduced morphine exposure (1.0 and 3.7 versus 8.7 µg · h/L, p < 0.001) as compared with normal metabolisers. Our final model leads the way in implementing model-informed precision dosing in codeine therapy and identifies the use of genetic testing as an integral component in the effort to implement rational pharmacotherapy with codeine.

Unveiling the Biological Effects and Pharmacological Properties of Sorbifolin: a Comprehensive Review.

Bioactive phytochemicals act as important factors with preventive and therapeutic potential in the pathogenesis of several disorders, often related to oxidative stress. Many dietary plant secondary metabolites could lower these conditions. Sorbifolin is one of these metabolites. This work is the first review of sorbifolin, a flavone detected in various plant matrices as a major compound. The present study discussed the natural sources, extraction, purification, quantification, and assessment of the biological activities of sorbifolin. Several databases including Google Scholar, Web of Sciences, and Science-Direct were consulted for relevant English articles related to sorbifolin, the phytochemical profiles of several medicinal plants containing this compound, and its biological activities, such as antioxidant, anticancer, antimicrobial, anti-inflammatory, and antidiabetic. The positive in vitro and in silico outcomes reported in the literature should be followed by additional in vivo and clinical investigations to further research the mechanisms of action, pharmacokinetic/pharmacodynamic activities, toxicological effects, pharmacological properties, and therapeutic potential of sorbifolin.

Autoinduction-Based Quantification of In Situ TGF-β Activity in Native and Engineered Cartilage.

Transforming growth factor beta (TGF-β) is a potent growth factor that regulates the homeostasis of native cartilage and is administered as an anabolic supplement for engineered cartilage growth. The quantification of TGF-β activity in live tissues in situ remains a significant challenge, as conventional activity assessments (e.g., Western blotting of intracellular signaling molecules or reporter cell assays) are unable to measure absolute levels of TGF-β activity in three-dimensional tissues. In this study, we develop a quantification platform established on TGF-β's autoinduction response, whereby active TGF-β (aTGF-β) signaling in cells induces their biosynthesis and secretion of new TGF-β in its latent form (LTGF-β). As such, cell-secreted LTGF-β can serve as a robust, non-destructive, label-free biomarker for quantifying in situ activity of TGF-β in live cartilage tissues. Here, we detect LTGF-β1 secretion levels for bovine native tissue explants and engineered tissue constructs treated with varying doses of media-supplemented aTGF-β3 using an isoform-specific ELISA. We demonstrate that: 1) LTGF-β secretion levels increase proportionally to aTGF-β exposure, reaching 7.4- and 6.6-fold increases in native and engineered cartilage, respectively; 2) synthesized LTGF-β exhibits low retention in both native and engineered cartilage tissue; and 3) secreted LTGF-β is stable in conditioned media for 2 weeks, thus enabling a reliable biological standard curve between LTGF-β secretion and exposed TGF-β activity. Accordingly, we perform quantifications of TGF-β activity in bovine native cartilage, demonstrating up to 0.59 ng/mL in response to physiological dynamic loading. We further quantify the in situ TGF-β activity in aTGF-β-conjugated scaffolds for engineered tissue, which exhibits 1.81 ng/mL of TGF-β activity as a result of a nominal 3 μg/mL loading dose. Overall, cell-secreted LTGF-β can serve as a robust biomarker to quantify in situ activity of TGF-β in live cartilage tissue and can be potentially applied for a wide range of applications, including multiple tissue types and tissue engineering platforms with different cell populations and scaffolds.

Analysis of good practices of social impact measurement: the social balance sheet developed by the Spanish network of the social and solidarity economy

Purpose This study aims to analyse the good practices developed by the Spanish Social and Solidarity Network (one of the leading networks in the country) regarding its social balance sheet, created under the perspective of social and solidarity principles. This social balance sheet is implemented by its members for their self-evaluation. Design/methodology/approach By using a qualitative methodology based on triangulation, which includes a semi-structured interview with the manager of the social audit commission, the main good practices associated with this initiative of social impact measurement are attained. Moreover, two organisations implementing the social balance sheet have been analysed to obtain a complete perspective of the tool and the process developed by the network. Findings The main findings include: the social balance sheet dimensions are based on the social and solidarity principles, which enable companies to analyse the compliance of their activity with their philosophy; cooperation is crucial for the success of the tool; the efforts made by the network involve a range of different behaviours, all shared and published on social networks and the internet for free to diffuse the good practices of social impact measurement. Originality/value This research contributes to the literature on social entrepreneurship and social impact measurement by analysing the good practices regarding a social balance sheet, designed under the social and solidarity philosophy, whose procedure can serve as a reference for any other representative institutions or practitioners. From this view, this initiative can be considered a social innovation itself since it constitutes a step towards transforming society through the quantification of business activity from a more social perspective and through a process of participative governance by all the stakeholders involved.

Propagation of underwater wave groups in a compressible ocean coupled with an elastic seafloor

The overall system of interest is an infinite half-space in which a compressible ocean is the top layer and an elastic seafloor (together with the crust beneath) forms a semi-infinite bottom layer. Whereas water-column compression waves and seafloor waves individually have received considerable attention, not much is known about their propagation as groups. This work utilizes the group behaviour of these waves to derive energy balance relations for wavenumber spectra for wave groups propagating through a mildly non-uniform water-column–seafloor system. Dispersion relations for the coupled system are derived using known kinematic and kinetic conditions at the interface, and free and forced wave solutions for the wavenumber spectra are obtained, with particular attention to the case when certain frequency–wavenumber combinations in the forcing excite the two-media system into resonance. Wavenumber spectra predicted using the theory for mildly non-uniform media are found to be close to those predicted assuming uniform media, though the effect of non-uniformity becomes more noticeable as the groups propagate farther from the generation area. Here, nonlinear interactions among stationary, random multi-directional surface-wave fields provide the forcing for groups of compression waves in the water and surface waves on the seafloor. The formulation includes the cumulative effect of multiple generation areas along the group propagation direction. Comparisons with observational data from a sensor array in the Atlantic Ocean indicate that the theory can be applied to reconstruct plausible combinations of generation areas and interaction times that are consistent with the measured data, for deriving approximate predictions at down-wave distances along the group propagation directions. Implications of this and other findings are discussed for (i) the potential for energy conversion from water-column compression waves on the seafloor, (ii) tracking of tropical cyclones from the seafloor, and (iii) quantification and comparative assessment of low-frequency mid-ocean ambient noise and microseism activity.

DNA polymerase mediated CRISPR/Cas12a trans-cleavage for dual-mode quantification of uracil DNA glycosylase activity

Since an unusual expression of uracil-DNA glycosylase (UDG) is often associated with the pathogenesis of numerous disorders, the detection of UDG activity is regarded as a promising application in disease diagnosis. Here, we develop a DNA polymerase-mediated CRISPR/Cas12a trans cleavage strategy, which can achieve dual-mode determination of UDG activity. By introducing a hairpin DNA probe containing a single uracil base, the probe undergoes specific cleavage and elongation under the existence of UDG only, thus activating the trans cleavage of ssDNA regardless of its length and sequence. To accommodate different detection modes, the ssDNA was further modified by fluorophore-quencher pairs or designed for connecting magnetic microparticles (MMPs) and polystyrene microparticles (PMPs). Finally, the UDG activity is quantified by fluorescence signal and microparticle accumulation length on a microfluidic chip visible to the naked eye. This strategy provides a detectable minimum UDG concentration of 0.00047 U/mL for fluorescent mode and 0.0048 U/mL for microfluidic mode. Furthermore, we performed the UDG inhibition test and detected UDG activity in cell lysates, suggesting its potential for inhibitor screening and detection of UDG in biological samples.

Facile quantitative diagnostic testing for neutralizing antibodies against Chikungunya virus

BackgroundViral neutralization (NT) assays can be used to determine the immune status of patients or assess the potency of candidate vaccines or therapeutic monoclonal antibodies (mAbs). Focus reduction neutralization test (FRNT) is a conventional neutralization test (cVNT) with superior specificity for measurement of neutralizing antibodies against a specific virus. Unfortunately, the application of FRNT to the chikungunya virus (CHIKV) involves a highly pathogenic bio-agent requiring biosafety level 3 (BSL3) facilities, which inevitably imposes high costs and limits accessibility. In this study, we evaluated a safe surrogate virus neutralization test (sVNT) that uses novel CHIKV replicon particles (VRPs) expressing eGFP and luciferase (Luc) to enable the rapid detection and quantification of neutralizing activity in clinical human serum samples.MethodsThis unmatched case-control validation study used serum samples from laboratory-confirmed cases of CHIKV (n = 19), dengue virus (DENV; n = 9), Japanese encephalitis virus (JEV; n = 5), and normal individuals (n = 20). We evaluated the effectiveness of sVNT, based on mosquito cell-derived CHIK VRPs (mos-CHIK VRPs), in detecting (eGFP) and quantifying (Luc) neutralizing activity, considering specificity, sensitivity, and reproducibility. We conducted correlation analysis between the proposed rapid method (20 h) versus FRNT assay (72 h). We also investigated the correlation between sVNT and FRNT in NT titrations in terms of Pearson’s correlation coefficient (r) and sigmoidal curve fitting.ResultsIn NT screening assays, sVNT-eGFP screening achieved sensitivity and specificity of 100%. In quantitative neutralization assays, we observed a Pearson’s correlation coefficient of 0.83 for NT50 values between sVNT-Luc and FRNT.ConclusionsFacile VRP-based sVNT within 24 h proved highly reliable in the identification and quantification of neutralizing activity against CHIKV in clinical serum samples.

AI-enabled CT-guided end-to-end quantification of total cardiac activity in 18FDG cardiac PET/CT for detection of cardiac sarcoidosis.

[18F]-fluorodeoxyglucose ([18F]FDG) positron emission tomography (PET) plays a central role in diagnosing and managing cardiac sarcoidosis. We propose a fully automated pipeline for quantification of [18F]FDG PET activity using deep learning (DL) segmentation of cardiac chambers on computed tomography (CT) attenuation maps and evaluate several quantitative approaches based on this framework. We included consecutive patients undergoing [18F]FDG PET/CT for suspected cardiac sarcoidosis. DL segmented left atrium, left ventricular(LV), right atrium, right ventricle, aorta, LV myocardium, and lungs from CT attenuation scans. CT-defined anatomical regions were applied to [18F]FDG PET images automatically to target to background ratio (TBR), volume of inflammation (VOI) and cardiometabolic activity (CMA) using full sized and shrunk segmentations. A total of 69 patients were included, with mean age of 56.1 ± 13.4 and cardiac sarcoidosis present in 29 (42%). CMA had the highest prediction performance (area under the receiver operating characteristic curve [AUC] 0.919, 95% confidence interval [CI] 0.858 - 0.980) followed by VOI (AUC 0.903, 95% CI 0.834 - 0.971), TBR (AUC 0.891, 95% CI 0.819 - 0.964), and maximum standardized uptake value (AUC 0.812, 95% CI 0.701 - 0.923). Abnormal CMA (≥1) had a sensitivity of 100% and specificity 65% for cardiac sarcoidosis. Lung quantification was able to identify patients with pulmonary abnormalities. We demonstrate that fully automated volumetric quantification of [18F]FDG PET for cardiac sarcoidosis based on CT attenuation map-derived volumetry is feasible, rapid, and has high prediction performance. This approach provides objective measurements of cardiac inflammation with consistent definition of myocardium and background region.

Changes in the fruits and seeds morphometric, germination, phytochemicals content, and antioxidant capacity in seed ripening of Echinocereus stramineus

Within the cacti family, most studies have focused on the physicochemical characterization of the stems, fruits, and seeds from Mammillaria, Opuntia, Hylocereus, and Stenocereus genera. However, few information is focused on the morphological and physicochemical characterization of the stems, fruits, and seeds of other genera widely distributed in the arid and semiarid zones of the Chihuahuan desert, such as the genus Echinocereus. The objective of this study was to analyze the fruit morphology, morphometry, germination process, and phytochemical content in four stages of seed in fruit ripening from E. stramineus. Morphometric parameters were measured by picking ten fruits and sixty seeds for each ripening stage. The characterization of the germination process included the germination percentage, mean germination time, germination speed, and mean germination speed which were computed 21 days after germination. The quantification of total phenols, flavonoids, tannins, reducing sugars, protein, and antioxidant activity of seeds was determined using colorimetric approaches under basal conditions. The morphometric results revealed a negative correlation between the fruit ripening stage and total mass (r= -0.980, p = 0.020), shell mass (r= -0.986, p = 0.014), pulp mass (r = -0.979, p = 0.021), fruit length (r = -0.978, p = 0.022), fruit width (r = -0.968, p = 0.032) and fruit area (r = -0.960, p = 0.04). The germination characterization process showed a negative association between the fruit ripening stage and the seed germination percentage (r = -0.979, p = 0.021) and between the seed mean germination time and the mean germination speed (r= -0.986, p = 0.014). The content of flavonoids, reducing sugars, proteins, and antioxidant capacity showed significant differences among the four stages of fruit ripening; however, no association was found between seed phytochemical content and the ripening stage. This study provides the first data on seed phytochemicals and information on the germination process of E. stramineus seeds.

Imaging Neural Activity in Intact, Semirestrained Drosophila Larvae.

The Drosophila larval nerve cord, which is the equivalent of the vertebrate spinal cord, houses the circuits required to process somatosensory stimuli (e.g., tactile, temperature, vibration, and self-movement) and generate the patterned muscle contractions underlying movement and behavior. Within this complex structure reside many cell types and cellular processes, making it difficult to experimentally access, when compared to peripheral parts of the nervous system (i.e., primary sensory neuron dendrites, motor neuron axons and synapses, and muscles). Additionally, the neurons in the larval nerve cord have small cell bodies, precluding traditional electrophysiological approaches. As such, the function of neurons in the nerve cord is less well studied than other parts of the nervous system, severely limiting our understanding of how larvae process sensory information and generate movement. Ca2+-sensitive fluorescent proteins enable the study of neuronal activity in live, genetically tractable animals, even those with small neuronal cell bodies. In addition, live imaging of neurons within the nerve cord in whole, intact animals is possible because larvae are translucent, and the use of intact animals allows for the peripheral sensory neuron circuits to remain intact. Ca2+-sensitive fluorescent proteins increase their fluorescence when voltage-gated Ca2+ channels are opened in depolarized neurons. Here, we describe an assay where a Ca2+-sensitive fluorescent protein (GCaMP6m) is expressed under the control of a GAL4 driver in a subset of neurons that reside in a circuit for vibration sensation. External vibration (sound) stimulates sensory neurons that activate the cells expressing the Ca2+-sensitive fluorescent protein. Visualization of the calcium-induced fluorescent signal with microscopy allows for quantification of neuronal activity.

Development of Surface EMG for Gait Analysis and Rehabilitation of Hemiparetic Patients.

The quantification of electromyographic activity using surface electrodes is invaluable for understanding gait disorders in patients with central nervous system lesions. We propose to evaluate a commercially available low-cost system compared to a reference system in participants with stroke-related movement disorders in functional situations. Three hemiparetic participants performed three functional tasks: two treadmill walks at different speeds and a sit-to-stand test. The vastus lateralis and gastrocnemius medialis muscles were equipped with two EMG sensors. The comparison between the two EMG systems was based on 883 identified cycles. Spearman's correlation coefficients (SCs), linear correlation coefficients (LCCs), and cross-correlation coefficients (CCCs) were calculated. The main results indicate good to very good similarity of the EMG signals collected from the two tested sEMG systems. In the comfortable-walking condition, an SC of 0.894 ± 0.091 and an LCC of 0.909 ± 0.094 were noted. In the fast-walking condition, an SC of 0.918 ± 0.064 and an LCC of 0.935 ± 0.056 were observed. For the 1 min sit-to-stand test, an SC of 0.880 ± 0.058 and an LCC of 0.881 ± 0.065 were noted. This study demonstrates good to very good similarity between the two sEMG systems, enabling the analysis of muscle activity during functional tasks.

Quantification and prediction of furnace cylinder activity based on LASSO-GWO-CNN hybrid model

Blast furnace smelting environments are complex and characterised by multi-scale, non-linear and strong coupling, and it is often difficult for operators to detect changes in cylinder activity in a timely manner. Quantifying the cylinder activity is the basis for monitoring and improving the cylinder activity. Aiming at the traditional method of quantifying the cylinder activity, which relies heavily on a single empirical formula and is difficult to realise the prediction effect of the cylinder activity, this paper proposes a new method of quantitatively characterising the cylinder activity by combining the core dead material column temperature index and the dead material pile cleanliness index, which comprehensively takes into account the thermodynamic and kinetic characteristics of the cylinder, and is able to more accurately and comprehensively assess the state of the cylinder activity. Firstly, a new quantitative cylinder activity formula is set according to the combined index, in which the weights of the indexes will be gradually adjusted in the iterative process of the model, and the entropy weighting method is adopted for its operation; a hierarchical affiliation function is established, and the calculation of the affiliation is used to judge the degree of contribution of the new indexes to the active state of the furnace cylinder with different weights in order to assist in finalising the final weights of the indexes, and the final weights of the indexes are then validated through the analysis of the calculation examples. Afterwards, the rationality of the final weights and the effectiveness of the proposed quantisation method are verified through case analysis. Finally, the LASSO algorithm is used to filter the feature variables, and the Grey Wolf Optimisation (GWO) optimised Convolutional Neural Network (CNN) based cylinder activity prediction model is proposed. According to the results of the comparative validation between five different prediction models, the prediction accuracy of the GWO-CNN model reaches more than 90%, which indicates that the model has a higher prediction performance in predicting the furnace cylinder activity.

A Gaussia luciferase reporter assay for the evaluation of coronavirus Nsp5/3CLpro activity

Human coronaviruses (hCoVs) infect millions of people every year. Among these, MERS, SARS-CoV-1, and SARS-CoV-2 caused significant morbidity and mortality and their emergence highlights the risk of possible future coronavirus outbreaks. Therefore, broadly-active anti-coronavirus drugs are needed. Pharmacological inhibition of the hCoV protease Nsp5 (3CLpro) is clinically beneficial as shown by the wide and effective use of Paxlovid (nirmatrelvir, ritonavir). However, further treatment options are required due to the risk of drug resistance. To facilitate the assessment of coronavirus protease function and its pharmacological inhibition, we developed an assay allowing rapid and reliable quantification of Nsp5 activity under biosafety level 1 conditions. It is based on an ACE2-Gal4 transcription factor fusion protein separated by a Nsp5 recognition site. Cleavage by Nsp5 releases the Gal4 transcription factor, which then induces the expression of Gaussia luciferase. Our assay is compatible with Nsp5 proteases from all hCoVs and allows simultaneous measurement of inhibitory and cytotoxic effects of the tested compounds. Proof-of-concept measurements confirmed that nirmatrelvir, GC376 and lopinavir inhibit SARS-CoV-2 Nsp5 function. Furthermore, the assay accurately predicted the impact of Nsp5 mutations on catalytic activity and inhibitor sensitivity. Overall, the reporter assay is suitable for evaluating viral protease activity.

An Automated pre-Dilution Setup for Von Willebrand Factor Activity Assays.

Accurate quantification of von Willebrand factor ristocetin cofactor activity (VWF:RCo) is critical for the diagnosis and classification of von Willebrand disease, the most common hereditary and acquired bleeding disorder in humans. Moreover, it is important to accurately assess the function of von Willebrand factor (VWF) concentrates within the pharmaceutical industry to provide consistent and high-quality biopharmaceuticals. Although the performance of VWF:RCo assay has been improved by using coagulation analyzers, which are specialized devices for blood and blood plasma samples, scientists still report a high degree of intra- and inter-assay variation in clinical laboratories. Moreover, high, manual sample dilutions are required for VWF:RCo determination of VWF concentrates within the pharmaceutical industry, which are a major source for assay imprecision. For the first time, we present a precise and accurate method to determine VWF:RCo, where all critical pipetting and mixing steps are automated. A pre-dilution setup was established on CyBio FeliX (Analytik-Jena) liquid handling system, and an adapted VWF:RCo method on BCS-XP analyzer (Siemens) is used. The automated pre-dilution method was executed on three different, most frequently used coagulation analyzers and compared to manual pre-dilutions performed by an experienced operator. Comparative sample testing revealed a similar assay precision (coefficient of variation = 5.9% automated, 3.1% manual pre-dilution) and no significant differences between the automated approach and manual dilutions of an expert in this method. While no outliers were generated with the automated procedure, the manual pre-dilution resulted in an error rate of 8.3%. Overall, this operator-independent protocol enables standardization and offers an efficient way of fully automating VWF activity assays, while maintaining the precision and accuracy of an expert analyst. Key features • Automated pre-dilution setup for von Willebrand factor concentrates of various natures. • Combination of a liquid handling system (CyBio FeliX) with a coagulation analyzer (BCS-XP). • Simplifies method transfer to other laboratories. • Basic training for CyBio FeliX and BCS-XP is required. Graphical overview VWF:RCo assay principle and measurement setup. Platelets (yellow ellipsoids) with negative surface charge (- - -) are treated with formaldehyde, which partly denatures the cell surface and thus stabilizes platelets for use as assay reagents. Stabilized platelets (dark-yellow-framed yellow ellipsoids) are then brought in contact with ristocetin A (chemical structure shown; black dots), which binds to the platelet surface and facilitates binding of VWF (green circles). The graphs show an example of quantitative determination of platelet agglutination by measurement of light transmission, where increasing amounts of VWF increase light transmission over time. The photo in the left-bottom corner shows the CyBio FeliX setup for VWF sample dilution and the photo in the right-bottom corner displays the BCS-XP system, which is used for VWF:RCo measurements.

Investigation of musculoskeletal symptoms, work postures, quantification of muscle activity, and estimation of grip/push forces among sonographers

AbstractDue to the physical nature of their work, sonographers are exposed to many musculoskeletal disorder risk factors, including awkward posture, repetitive movements, forceful manual exertion, and static muscle contractions, especially in the upper limbs. The current study is an investigation of musculoskeletal disorders among sonographers, caused by various occupational risk factors via different sonographic scan types. During the first phase of this study, the musculoskeletal symptoms and work postures of 29 subjects were investigated. During the second phase, muscle activity was quantified, and grip/push forces were estimated using the data obtained from 10 volunteer sonographers. 82% of sonographers experienced musculoskeletal symptoms. Based on the final scores and action levels obtained via rapid upper limb assessment, while performing scans of left regions; ergonomic changes and interventions were found necessary, to relieve stress on the sonographer's body. The results of muscular activity per muscle and scan type, showed that the mean muscle activity of the middle deltoid muscle was significantly higher during the right abdominal scan (17.64% maximum voluntary contraction [MVC]), compared to those of thyroid (12.54% MVC) and left abdominal (7.32% MVC) scans. Additionally, mean grip and push forces during both abdominal scans were significantly higher than those during the thyroid scan. Despite an injury risk during all scans, risk factor impact was different among scan types. This groundbreaking study represents the first that captures and measures both grip and push forces simultaneously, which may prove helpful while investigating corrective interventions or optimizing design of sonography robots and ergonomic probes in future studies.

Imaging of Ulcerative Colitis: The Role of Diffusion-Weighted Magnetic Resonance Imaging.

Magnetic resonance imaging (MRI) has emerged as a promising and appealing alternative to endoscopy in the objective assessment of patients with inflammatory bowel disease (IBD). Diffusion-weighted imaging (DWI) is a specialized imaging technique that enables the mapping of water molecule diffusion within biological tissues, eliminating the need for intravenous gadolinium contrast injection. It is expanding the capability of traditional MRI sequences in Ulcerative Colitis (UC). Recently, there has been growing interest in the application of intravoxel incoherent motion (IVIM) imaging in the field of IBD. This technique combines diffusion and perfusion information, making it a valuable tool for assessing IBD treatment response. Previous studies have extensively studied the use of DWI techniques for evaluating the severity of activity in IBD. However, the majority of these studies have primarily focused on Crohn's disease (CD), with only a limited number of reports specifically examining UC. Therefore, this review briefly introduces the basics of DWI and IVIM imaging and conducts a review of relevant studies that have investigated its application in UC to show whether these techniques are useful techniques for evaluating patients with UC in terms of detection, characterization, and quantification of disease activity. Through the extensive literature survey, most of these studies indicate that DWI proves valuable in the differential diagnosis of UC and could be used as an effective modality for staging UC.

Single G-quadruplex-based fluorescence method for the uracil-DNA glycosylase inhibitor screening

Uracil-DNA glycosylase (UDG) plays a pivotal role in the base repair system. Through bioinformatics, we found that the expression of the UDG enzyme in many cancer cells is increased, and its high expression is not conducive to the prognosis of lung cancer patients. The development of analytical techniques for the quantification of UDG activity and the identification of UDG inhibitors is of paramount importance. We found that when the T base in the G4 loop region mutated to uracil, the G4 structure was not disrupted and still retained the characteristics of a G4 structure (emitting strong fluorescence after binding with ThT (Thioflavin T). Inspired by this phenomenon, we developed a detection method for UDG and its inhibitors utilizing a single DNA strand engineered to form a G-quadruplex structure, containing uracil residues within the loop region, designated as G4-dU. The inclusion of uracil-DNA glycosylase (UDG) in the assay environment induces the removal of uracil, resulting in the formation of apurinic sites (AP) within the G4-dU sequence. Subsequent thermal denaturation leads to strand cleavage at AP sites, precluding the reformation of the G-quadruplex configuration and abrogating fluorescence emission. The detection process in this study can be completed in only 30 min to 1 h, offers a straightforward, expedient, and efficacious modality for assessing UDG activity and UDG inhibitor potency, thereby facilitating the discovery of novel therapeutic agents for cancer treatment.

Quantification of macrophage activity in knee synovial tissue using [18F]FEPPA positron emission tomography

ObjectiveInflammation in knee osteoarthritis (OA) is mediated primarily by synovial tissue macrophages, but non-invasive measurement of macrophage activity in vivo is a challenge. Activated macrophages markedly increase expression of the translocator protein (TSPO). In the context of other diseases TSPO-PET using the [18F]FEPPA tracer (binding to TSPO) has been reported to be an effective method for imaging macrophages in vivo. The goal of this study was to validate the use of [18F]FEPPA PET radiotracer to accurately measure macrophage activation in knee synovial tissue. DesignTen participants with late-stage OA scheduled for knee replacement surgery were recruited. Prior to surgery, 5 of the participants underwent a clinical [18F]FEPPA PET/MRI scan and the standard uptake value (SUV) in the suprapatellar synovial region was calculated. Suprapatellar synovial tissue samples were collected from all participants at the time of surgery and were imaged ex vivo with [18F]FEPPA autoradiography. Tracer uptake was compared to TSPO antibody immunostaining in serial tissue sections. The correlation between the [18F]FEPPA uptake and gold standard TSPO fluorescent intensity was measured. RESULTSThe autoradiography [18F]FEPPA signal in the synovial lining was correlated with the TSPO signal in the. lining macrophages measured by immunohistochemistry (r = 0.81, p = 0.0040, CI [0.37, 0.95]). Similarly, PET/MRI scans demonstrated similar correlation between SUV in synovial tissues in vivo and in situ TSPO signal in lining macrophages {r = 0.90, p = 0.083, CI [0.086, 0.99]) Conclusion[18F]FEPPA uptake in knee synovial tissue is strongly correlated to the expression of TSPO in synovial lining macrophages, suggesting that clinical [18F]FEPPA PET/MRI may be a valid measure of true macrophage activity in synovial tissues in knee OA. [18F]FEPPA may be an effective tool to assess macrophage activation both ex vivo and in vivo, and should be investigated further to assess its performance and measurement properties in different clinical contexts of knee OA including disease states and responses to treatment.