Quantification Of Activity Research Articles

Increased neutrophil elastase in affected lobes of bronchiectasis and correlation of its levels between sputum and bronchial lavage fluid.

Neutrophil elastase (NE) has been proposed as a potential biomarker for evaluating the severity and prognosis of bronchiectasis. This study aimed to compare bronchial lavage quantification of NE levels and activities with those of sputum. A cross-sectional study was conducted in which 24 Vietnamese adults with bronchiectasis were enrolled from June 2023 to August 2023. All participants underwent bronchoscopy to collect bronchial lavage fluid (BLF) from two bronchial locations: one in the region with the greatest bronchial dilatation and one in the normal bronchi or in patients with all lobes affected, the least abnormal lobe (ABLF and NBLF, respectively). Spontaneously expectorated sputum was also collected. Out of 24 cases, the prevalence of mild, moderate and severe bronchiectasis was 14/24 (58.4%), 5/24 (20.8%), and 5/24 (20.8%), respectively. NE concentration and activity were significantly higher in sputum and ABLF than in NBLF (p < 0.001). Sputum and ABLF were highly correlated (r = 0.841, p < 0.001) with no significant difference in NE activity between sputum and ABLF. Higher levels of NE activity were seen in more severe bronchiectasis than in mild bronchiectasis in all samples but were only statistically significant for NE activity in sputum (r = 0.418, p = 0.042). NE activity and concentration are elevated in areas of the lung most affected by bronchiectasis. Sputum is a valid surrogate of pulmonary NE levels, as they correlate strongly with ABLF and confirm in a Vietnamese population the relationship between NE activity and disease severity.

Prymnesium parvum: Correlation between lytic activity and prymnesins content, both increasing with phosphorus-depletion and lower cell density

The haptophyte Prymnesium parvum is a widespread microalgae associated with fish kill events. It is a mixotrophic alga, which combines different trophic modes of nutrient and energy acquisition, in part relying on the consumption of prey, a process in which its toxins, the prymnesins (PRMs) are suspected to be involved. Investigating toxicity, and more specifically quantifying PRMs, is challenging because of a lack of standards. In this study, two experiments were conducted to better understand the effects of phosphorus sufficiency and deficiency, growth phase and cell density on lytic activity and PRMs content (intra- and extracellular) of one strain of P. parvum (CCAP 946/6). In the first experiment, P. parvum cultures were grown with or without addition of phosphorus as either inorganic nutrient and/or an algal prey (i.e. the cryptophyte Teleaulax amphioxeia). In the second experiment, a phosphorus-starved culture of P. parvum was inoculated at three initial cell densities in phosphorus-deficient medium. Detection and quantification of phagotrophic activity and toxin content were performed using flow cytometry and liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS), respectively. In addition, to assess the lytic activity of P. parvum, microalgal bioassays were performed on the supernatant towards a competitor, T. amphioxeia. Our results show for the first time a positive correlation between PRMs content and lytic effect on T. amphioxeia. Interestingly, predation had no effect on the PRMs, neither intra- nor extracellular forms, and the toxin content was mainly influenced by the depletion of inorganic phosphorus. Secondly and unexpectedly, lytic effect and PRMs content were higher at lower cell densities in phosphorus-deficient medium. These data provide new insights into the factors promoting the toxicity of P. parvum which may support the development of standards that could help to understand the role and mode of action of these toxins, and their impact on ecosystems.

On-site visual quantification of alkaline phosphatase activity in cells using a smartphone-based approach.

Alkaline phosphatase (ALP) is a critical biomarker associated with various physiological and pathological processes, making its detection essential for disease diagnosis and biomedical research. In this study, we developed a novel, simple, and portable visual quantification method for ALP activity in cells using an efficient Cu0.9Zn0.1S nanomaterial with peroxidase-like properties, integrated into a smartphone-based platform for enhanced usability. The Cu0.9Zn0.1S nanomaterial catalyzes the breakdown of H₂O₂, generating ·OH radicals that oxidize the colorless substrate TMB into blue oxTMB, which is subsequently reduced back to TMB by ascorbic acid (AA). We employed an indirect colorimetric approach for ALP detection, leveraging ALP's ability to catalyze the dephosphorylation of l-ascorbic acid-2-phosphate (AAP) to produce AA. This method enabled highly sensitive and precise quantification of ALP with a detection limit of 0.47 mU/L and a linear range from 0.001 to 100 U/L. The established smartphone platform not only facilitated real-time data processing but also transformed the detection system into a portable and accessible laboratory. The method was successfully applied to detect ALP in various cancer cell lines, with the highest levels found in HepG2 cells. Moreover, the results were consistent with those obtained using traditional kit-based methods, highlighting the accuracy and reliability of our approach. The system was also applied to evaluate ALP inhibitors, demonstrating its versatility in biomedical applications. This innovative, cost-effective, and highly sensitive colorimetric sensing strategy offers immense potential for clinical diagnostics, cancer research, and inhibitor screening, particularly in on-site, resource-limited, or emergency scenarios.

Single-molecule quantification of photoredox activities and dynamics at the nanoscale on multi-faceted 2D materials

Single-molecule quantification of photoredox activities and dynamics at the nanoscale on multi-faceted 2D materials

Explainable Machine Learning-Based Approach to Identify People at Risk of Diabetes Using Physical Activity Monitoring

Objective: This study aimed to investigate the utilization of patterns derived from physical activity monitoring (PAM) for the identification of individuals at risk of type 2 diabetes mellitus (T2DM) through an at-home screening approach employing machine learning techniques. Methods: Data from the 2011–2014 National Health and Nutrition Examination Survey (NHANES) were scrutinized, focusing on the PAM component. The primary objective involved the identification of diabetes, characterized by an HbA1c ≥ 6.5% (48 mmol/mol), while the secondary objective included individuals with prediabetes, defined by an HbA1c ≥ 5.7% (39 mmol/mol). Features derived from PAM, along with age, were utilized as inputs for an XGBoost classification model. SHapley Additive exPlanations (SHAP) was employed to enhance the interpretability of the models. Results: The study included 7532 subjects with both PAM and HbA1c data. The model, which solely included PAM features, had a test dataset ROC-AUC of 0.74 (95% CI = 0.72–0.76). When integrating the PAM features with age, the model’s ROC-AUC increased to 0.79 (95% CI = 0.78–0.80) in the test dataset. When addressing the secondary target of prediabetes, the XGBoost model exhibited a test dataset ROC-AUC of 0.80 [95% CI; 0.79–0.81]. Conclusions: The objective quantification of physical activity through PAM yields valuable information that can be employed in the identification of individuals with undiagnosed diabetes and prediabetes.

Selection of GABA-Producing Lactic Acid Bacteria Strains by Polymerase Chain Reaction Using Novel gadB and gadC Multispecies Primers for the Development of New Functional Foods.

Gamma-aminobutyric acid (GABA) has been attributed to health-promoting properties and has received attention from the food industry as an attractive bioactive compound for the development of functional foods. Some lactic acid bacteria (LAB) produce GABA through a glutamate decarboxylase encoded by gadB and a glutamate/GABA antiporter encoded by gadC. In this study, we develop a molecular screening method based on a polymerase chain reaction able to detect those genes in different LAB species through the use of novel multispecies primers. PCR was performed in 92 LAB strains of six different species. The primer pair designed for gadB allowed its identification in Lactiplantibacillus plantarum, Lactococcus cremoris, Lactococcus lactis, Levilactobacillus brevis, Limosilactobacillus fermentum, and Limosilactobacillus reuteri strains. For gadC, two different primer pairs were designed for its detection in different species. Glutamate decarboxylase activity (GAD assay) and GABase enzymatic quantification were also assessed. Among those strains showing glutamate decarboxylase activity, 93.2% harbored the gadB gene, and those showing GABA production had the gadB gene and exhibited glutamate decarboxylase activity. PCR detection of gadB correlates strongly with GABA production and constitutes a good strategy for the selection of LAB with high yields (>18 mM) that could be used for the development of GABA-enriched functional foods.

Electrochemiluminescence Detecting and Imaging of Yeast Metabolism Indicated by Endogenetic Co-reactant.

Glycolysis, a pivotal step in yeast metabolism, plays an indispensable role as a carbohydrate utilization process crucial for cellular survival. Developing advanced technologies to elucidate this fundamental physiological process holds significant scientific implications. Electrochemiluminescence (ECL) imaging exhibits the advantage of negligible background interference and facilitates straightforward visualization, thereby conferring significant value in biomolecular observation. In this study, we present an ECL imaging method for investigating yeast metabolism by utilizing the endogenetic NADH as an efficient coreactant for ECL generation. The yeast glycolysis process drives the conversion of NAD+ to NADH, resulting in enhanced ECL response as well as the increased brightness of ECL images that can be used for quantification of yeast activity. There was a linear correlation between the reciprocal of both the gray value of ECL image and yeast concentration within the range of 6.25 × 106 - 6.25 × 108 CFU/mL. Due to the highly efficient coreactant behavior of NADH, our method demonstrated excellent selectivity with minimal interference. Furthermore, we employed this approach to investigate some toxic inhibitors on yeast metabolism, yielding reliable results. This ECL imaging method not only avoids the use of additional coreactants but also provides a sensitive and intuitive approach for monitoring yeast metabolism, demonstrating great potential in revealing various complex biological processes.

Oxygen Reduction Reaction on Pyridinic Nitrogen-Functionalized Carbon: Active Site Quantification and Effects of Lewis Basicity

Oxygen Reduction Reaction on Pyridinic Nitrogen-Functionalized Carbon: Active Site Quantification and Effects of Lewis Basicity

Quantification of gait characteristics and muscle activation in patients with chronic ankle instability during walking on sand: A randomized crossover trial.

Limited evidence exists regarding the movement control patterns of individuals with chronic ankle instability (CAI) during sand walking. This study aimed to analyze gait characteristics and muscle activation in patients with CAI while walking on sand. This study recruited 30 participants, including 15 patients with CAI and 15 healthy controls. Patients with CAI were selected based on having experienced at least a lateral ankle sprain within 12 months before recruitment. They also had to scoring ≤ 27 on the Cumberland ankle instability tool. This study was conducted using a randomized crossover trial. All participants were asked to walk three times each at their natural speed on both sand and paving blocks for a distance of 50m. There was at least a week between each trial. The lower limb gait kinematics, spatiotemporal, and muscle activity parameters while walking on sand and paving blocks were compared between both groups. Significant differences in knee angle were observed between 60.83% and 75.34% of the gait cycle during walking on sand in the patients with CAI. Regarding spatiotemporal parameters, cadence (P = .001) and stride length (P = .049) showed significant differences as the main effect. In the CAI group, the recruitment threshold for the peroneus longus muscle was significantly lower during walking on sand than on paving blocks (P < .001). In contrast, the motor unit action potential (MUAP) was significantly higher (P < .001). For the tibialis anterior muscle, the recruitment threshold and number of motor units were significantly lower during walking on sand than on paving blocks (P = .006 and P = .003, respectively), and the MUAP was significantly higher (P < .001). During walking on sand, patients with CAI exhibited increased knee flexion angle, stride length, and MUAP, along with decreased cadence and recruitment thresholds. These results suggest that sand can influence gait characteristics and muscle activation in patients with CAI. Our findings indicate that gait characteristics and muscle activation changes in patients with CAI could potentially benefit CAI rehabilitation.

Kinetic Profiling of Oxidoreductase-Mimicking Nanozymes: Impact of Multiple Activities, Chemical Transformations, and Colloidal Stability.

In contrast to homogeneous enzyme catalysis, nanozymes are nanosized heterogeneous catalysts that perform reactions on a rigid surface. This fundamental difference between enzymes and nanozymes is often overlooked in kinetic studies and practical applications. In this article, using 14 nanozymes of various compositions (core@shell, metal-organic frameworks, metal, and metal oxide nanoparticles), we systematically demonstrate that nontypical features of nanozymes, such as multiple catalytic activities, chemical transformations, and aggregation, need to be considered in nanozyme catalysis. Ignoring these features results in the inaccurate quantification of catalytic activity. Neglecting the multiple activities led to a six-time underestimation of Mn2O3 oxidation activity and mischaracterization of this material as a low-active peroxidase-mimicking nanozyme. Additionally, overlooking chemical stability during catalytic reactions led to the reporting of high peroxidase-mimicking activity for Au@Ag nanoparticles, which, in reality, exhibited no intrinsic activity and oxidized the substrate through the leakage of Ag+ ions. Ignoring the chemical stability of Au@Prussian Blue nanoparticles may lead to more than four times overestimation of peroxidase-mimicking activity after just 24 h of storage. Finally, disregarding the colloidal stability of nanozymes led to a five-time inaccuracy in catalytic activity. These findings underscore the necessity of optimizing procedures to account for these factors in nanozyme kinetic measurements, which will in turn ensure more reliable biosensors and the success of other practical applications.

Institutional constraints to higher education datafication: an English case study

AbstractStrong claims are made by both higher education policymakers and institutional leaders about the advantages that can be brought about through ‘datafication’ (i.e. the quantification of human activities through digital means). Nevertheless, we know relatively little about how university cultures and processes may impinge on the rolling out of data practices, particularly in relation to administrative data (rather than that collected through learning analytics and other forms of user-generated data). This article draws on a particular case study—of attempts to widen participation to ‘sandwich courses’ in English higher education institutions—to argue that, in some areas of activity at least, institutional constraints serve to limit datafication substantially, including insufficient time and infrastructure, the prioritisation of data required for other purposes (typically national assessment exercises) and the role of institutional cultures. Cognisant of various problems associated with quantification, the article nevertheless contends that the barriers to the effective use of metrics identified in this research may have a direct impact on institutions’ capacity to recognise and address social inequalities.

Design of a Bioluminescent Assay Platform for Quantitative Measurement of Histone Acetyltransferase Enzymatic Activity.

Protein acetylation and acylation are widespread post-translational modifications (PTMs) in eukaryotic and prokaryotic organisms. Histone acetyltransferase (HATs) enzymes catalyze the addition of short-chain acyl moieties to lysine residues on cellular proteins. Many HAT members are found to be dysregulated in human diseases, especially oncological processes. Screening potent and selective HAT inhibitors has promising application for therapeutic innovation. A biochemical assay for quantification of HAT activity utilizing luminescent output is highly desirable to improve upon limitations associated with the classic radiometric assay formats. Here we report the design of a bioluminescent technological platform for robust and sensitive quantification of HAT activity. This platform utilizes the metabolic enzyme acetyl-CoA synthetase 1 (ACS1) for a coupled reaction with firefly luciferase to generate luminescent signal relative to the HAT-catalyzed acetylation reaction. The biochemical assay was implemented in microtiter plate format and our results showed this assay sensitively detected catalytic activity of HAT enzyme p300, accurately measured its steady-state kinetic parameters of histone acetylation and measured the inhibitory potency of HAT inhibitor. This platform demonstrated excellent robustness, reproducibility, and signal-to-background ratios, with a screening window Z'=0.79. Our new bioluminescent design provides an alternative means for HAT enzymatic activity quantitation and HAT inhibitor screening.

Antioxidant activities of aqueous and 70% ethanol extracts of akar kuning (Arcangelisia flava (L.) Merr) stem using the DPPH method

Background: Numerous herbs can be used as natural exogenous antioxidants and clinically proven effective as antioxidants. Objective: The primary objective of this study is to assess the antioxidant activity of the stems of akar kuning (Arcangelisia flava (L.) Merr). Method: This plant was extracted with aqueous solution using the decoction method and 70 % ethanol using the maceration method. Antioxidant activity was evaluated using the TLC-Bioautography sprayed with DPPH reagent for the qualitative method and microplate assay with DPPH for the quantitative method. This research also identified the total phenol from each extract which was measured using the Folin-Ciocalteu method. Result: Based on phytochemical screening, each extract contained flavonoids, polyphenols, anthraquinones, terpenoids, and saponins. TLC-bioautography showed flavonoids, polyphenols, and anthraquinones were responsible for antioxidant activity. Quantification of antioxidant activity results of aqueous and 70% ethanol extracts of akar kuning (Arcangelisia flava (L.) Merr) showed IC50 value at 78.8 ± 1.2 µg/mL and 142.8± 2.9 µg/mL respectively. The content of total phenolic compounds was 80.544 ± 2.6 mg GAE/ g and 52.522 ± 1.9 mg GAE/g. Conclusion: As observed in the study, Arcangelisia flava (L.) Merr extracts from both extraction methods possess an antioxidant.

Assessment and role of physical activity as a factor affecting subjective perception of students' satisfaction

The paper aims to provide analysis of Slovak university students’ physical activity during state-ordered distance learning in the academic year 2021/2022 (the COVID-19 pandemic). The data were obtained in form of a questionnaire survey. In the analytical part, the focus is on weekly time that students devoted to regular exercise and quantification of physical activity versus subjective perception of selected physical and psychological satisfaction and health indicators. To meet the paper’s objectives, a two-sample t-test was used to test statistical significance of differences between men and women, and parametric and non-parametric correlation analysis methods. In terms of age structure majority of students falls into young adults’ category, in which physical activity has only a small positive effect on the subjective perception of satisfaction. Majority of students exercised on average up to 50 minutes per week, which is less than weekly physical activity during regular face-to-face lessons of Physical Education.

Unveiling the Biological Activities, Pharmacological Potential, and Health Benefits 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.

Nanozyme-Enzyme Cascade Reaction-Enhanced Ratiometric Fluorescence Immunosensing Platform for Sensitive and Accurate Detection of Ractopamine.

As the most widely used immunoassay, enzyme-linked immunosorbent assay (ELISA) relies on perishable enzymes and usually provides poor sensitivity and stability, limiting its application in detecting trace analytes in harsh environments. Herein, a new ratiometric fluorescence (RF) sensing platform enhanced by a nanozyme-enzyme cascade reaction composed of MnO2 nanosheets (MnO2 NSs) and alkaline phosphatase (ALP) was proposed. In this RF platform, the versatile MnO2 NSs worked as a robust oxidase-like nanozyme to catalyze nonfluorescent Amplex Red (AR) into fluorescent resorufin and as a quencher to quench the fluorescence of carbon dots (CDs). Given that ALP could catalyze ascorbic acid 2-phosphate (AAP) into ascorbic acid (AA), followed by AA reducing MnO2 NSs into Mn2+, resulting in the fluorescence of AR decrease but fluorescence of CD recovery simultaneously. The regulation of MnO2 NSs for the fluorescence intensity of AR (Em at 585 nm) and CDs (Em at 460 nm) could cascade with ALP-triggered reaction to construct an RF sensing platform based on RF signal (F585/F460) output. The proposed RF sensing platform could achieve a superior limit of detection (LOD) of 0.037 mU mL-1 for ALP activity quantification. Inspired by the excellent performance of the RF sensing platform, a sensitive and accurate RF ELISA for ractopamine (RAC) detection was established with an LOD of 0.029 ng mL-1, which was nearly 5.5 times more sensitive than traditional colorimetric ELISA. This RF sensing platform based on robust nanozyme-triggered enzymatic cascade signal amplification and excellent RF signal readout has offered a powerful and universal platform for RAC and other trace target detection.

Total antioxidant capacity status in non-obese adolescent females with PCOS: a cross-section study

BackgroundThe endocrine disease polycystic ovarian syndrome (PCOS) has a number of complications. Teenagers all over the globe are becoming increasingly concerned about the syndrome. It has been suggested that oxidative stress plays a significant role in the pathogenesis of PCOS.AimThe purpose of the present work was to estimate total antioxidant capacity (TAC) in non-obese adolescents and decide if the TAC could be a potential marker for the prediction and diagnosis of PCOS.MethodsSerum samples were collected to assess the levels of follicle stimulant hormone (FSH), luteinizing hormone (LH), serum lipids, and serum total antioxidant capacity (TAC) in 50 non-obese patients with PCOS and 50 controls; correlation analysis was made between serum lipid, hormonal parameters, and TAC.ResultsAdolescents with PCOS had significantly lower levels of TAC compared with controls and showed significant higher values of LH, FSH, and LH/FSH ratio. There were significant negative correlations between TAC levels with LH, FSH, and LH/FSH. Linear regression analysis revealed statistically significant correlations between TAC and LH and FSH levels in non-obese adolescents with PCOS.ConclusionSerum TAC level was diminished in non-obese PCOS patients and was related to increase of LH/FSH ratio. So, quantification of TAC activity in serum could be of value in assessment of cases at higher risk for development of PCOS, suggesting its possible role in regulating sex hormones and could have potential roles in the etiopathogenesis of PCOS in non-obese adolescent females.

Partial volume correction for Lu-177-PSMA SPECT

BackgroundThe limited spatial resolution in SPECT images leads to partial volume effect (PVE), degrading the subsequent dosimetric accuracy. We aim to quantitatively evaluate PVE and partial volume corrections (PVC), i.e., recovery coefficient (RC)-PVC (RC-PVC), reblurred Van-Cittert (RVC) and iterative Yang (IY), in 177Lu-PSMA-617 SPECT images.MethodsWe employed a geometrical cylindrical phantom containing five spheres (diameters ranging from 20 to 40 mm) and 40 XCAT phantoms with various anatomical variations and activity distributions. SIMIND Monte Carlo code was used to generate realistic noisy projections. In the clinical study, sequential quantitative SPECT/CT imaging at 4 time-points post 177Lu-PSMA-617 injections were analyzed for 10 patients. Iterative statistical reconstruction methods were used for reconstruction with attenuation, scatter and geometrical collimator detector response corrections, followed by post-filters. The RC-curves were fit based on the geometrical phantom study and applied for XCAT phantom and clinical study in RC-PVC. Matched and 0.5-2.0 voxels (2.54–10.16 mm) mismatched sphere masks were deployed in IY. The coefficient of variation (CoV) was measured on a uniform background on the geometrical phantom. RCs of spheres and mean absolute activity error (MAE) of kidneys and tumors were evaluated in simulation data, while the activity difference was evaluated in clinical data before and after PVC.ResultsIn the simulation study, the spheres experienced significant PVE, i.e., 0.26 RC and 0.70 RC for the 20 mm and 40 mm spheres, respectively. RVC and IY improved the RC of the 20 mm sphere to 0.37 and 0.75 and RC of the 40 mm sphere to 0.96 and 1.04. Mismatch in mask increased the activity error for all spheres in IY. RVC increased noise and caused Gibbs ringing artifacts. For XCAT phantoms, both RVC and IY performed comparably and were superior to RC-PVC in reducing the MAE of the kidneys. However, IY and RC-PVC outperformed RVC for tumors. The XCAT phantom study and clinical study showed a similar trend in the kidney and tumor activity differences between non-PVC and PVC.ConclusionsPVE greatly impacts activity quantification, especially for small objects. All PVC methods improve the quantification accuracy in 177Lu-PSMA SPECT.

EXTH-63. FOCUSED ULTRASOUND HYPERTHERMIA MEDIATED CONTROL OF THERMAL RESPONSIVE CAR T CELL ACTIVITY IN BREAST CANCER BRAIN METASTASIS

Abstract HER2-targeted therapies are promising treatment options for metastatic breast cancer and have improved the median overall survival. However, breast cancer brain metastasis (BCBM), observed in up to 50% of HER2-positive breast cancer patients, remains a clinical challenge. While Chimeric Antigen Receptor (CAR) T cell therapy is promising therapeutic strategy to address the unmet need in treating BCBM, improving its effectiveness and safety is critical for clinical translation. We hypothesized that spatially and temporally controlled activity of CAR-T cells with a thermal gene switch (TS) under MRI-guided focused ultrasound (MRgFUS) would potentiate anti-tumor responses in difficult-to-treat BCBM. To test our hypothesis, we developed a closed-loop controlled MR-thermometry (MRTI) based FUS system (MRgFUS) and αHER2 CAR-T cells engineered with TS expressing the reporter gene firefly luciferase (TS.Fluc) or bi-specific engager on NKG2D ligands (TS.BTE) upon MRgFUS-hyperthermia (41.5°C). In vivo quantification of TS activity from intratumorally delivered TS.Fluc αHER2 CAR-T cells in HER2+ BCBM mice model (HER2+ MDA-MB-468 in NSG mice with primary human CAR-T cells) followed by pulsed MRgFUS-hyperthermia resulted in a ~10-fold increase in luminescent activity compared to unheated mice. To assess longitudinal activation, we quantified TS activity with a second sonication 4 days post the initial FUS treatment. Prior to the second sonication, TS.Fluc expression had returned to baseline levels, while following it robust TS.Fluc expression was observed again, demonstrating that MRgFUS can activate engineered T cells with TS in brain tumors with high spatiotemporal control. Subsequently we investigated the therapeutic efficacy of intratumorally delivered TS.BTE αHER2 CAR T cells in mixed tumor model of heterogeneous HER2 MDA-MB-468 (75% HER2+) and observed significant tumor regression of HER2- tumor cells in the treated group compared to controls, demonstrating the potential of the proposed strategy to noninvasively drive the local production of key transgenes and potentiate anti-tumor responses in difficult-to-treat BCBM.

Quantitative evaluation of clinical pharmacy activities following implementation of an enhanced weekend service at a teaching hospital

Abstract Introduction Improving equity of staffing across seven-days is a strategic priority for the National Health Service (NHS).1 Development of enhanced weekend clinical pharmacy services presents opportunity to improve admission and discharge medicines reconciliation (MR), reduce medication error and increase patient counselling.2 At the study site, an enhanced weekend clinical pharmacy service was implemented in October 2021. The service was delivered 9am-5pm to all clinical specialities across 54 wards. Defined minimum staffing was 13 pharmacists and 2 medicines management pharmacy technicians (MMPT). Service specification included: clinical screen and order of medications; admission MR, review of high-risk patients (critical care patients and those prioritised by the weekday team); and discharge activities including patient counselling. Aim To evaluate provision of the enhanced weekend clinical pharmacy service post-implementation by quantification of shift fill and clinical pharmacy professional activities. Methods A quantitative study was conducted over two separate weekends one month and six months post-implementation (13th-14th November 2021 and 9-10th April 2022). Pharmacy professionals collected data using a paper activity tool validated for use at the study site. Interventions included rectification of medication error, medicines optimisation or consult. Data were collated in Microsoft Excel immediately after each study weekend. Simple descriptive statistics were performed in Microsoft Excel. The study was registered with KCH Pharmacy Research and Audit Group (07/2021), deemed service evaluation and ethics approval waived. Results Pharmacy professionals reviewed 1,056 patients over 3,645 occupied bed-days. Services were delivered by 13 pharmacists and 1 MMPT on weekend 1, 12 pharmacists and 1 MMPT on weekend 2. Data were collected for all wards on weekend 1; on weekend 2 activity data was not collected for five surgical wards (Saturday and Sunday), four post-acute medical wards (Saturday) and the acute medical unit (Sunday). Admission MR was conducted for 77% (317/412) of newly-admitted patients. Pharmacists recommended prescription amendment for 68% (217/317) of patients receiving admissions MR; affecting 517 medicines. The recorded intervention rate per patient reviewed was 0.64 (680/1056). Pharmacy professionals responded to 211 queries from healthcare professionals (HCPs), undertook 90 incidences of pro-active HCP education and counselled 92 patients/carers. Pharmacists prepared discharge medication lists or clinically screened discharge prescriptions for 205 patients. A total of 952 non-stock medicines were ordered and 313 patients own medicines checked for suitability for reuse. Discussion / Conclusion These findings add to existing evidence for seven-day clinical pharmacy services. Defined minimum staffing was not fulfilled with 90% shift fill rate. Clear benefit was demonstrated in terms of admission MR and resultant intervention, HCP education, discharge and counselling activities. The percentage of patients with a recommended prescription amendment following admission MR matched findings from a separate service evaluation during February 2021 of an enhanced weekend service introduced at our study site during the COVID-19 pandemic (68%).3 The intervention rate for reviewed patients was also comparable (0.64 compared with 0.57 during COVID-19).3 Limitations include the relatively short period of data collection at a single-site which limits generalisability of findings. There were some gaps in data collection and acceptance rate of interventions was not collected.