Following employers' criticisms and recent societal developments, policymakers and educators have called for students to develop a range of generic skills such as critical thinking ("twenty-first ...century skills"). So far, such skills have typically been assessed by student self-reports or with multiple-choice tests. An alternative approach is criterion-sampling measurement. This approach leads to developing performance assessments using "criterion" tasks, which are drawn from real-world situations in which students are being educated, both within and across academic or professional domains. One current project, iPAL (The international Performance Assessment of Learning), consolidates previous research and focuses on the next generation performance assessments. In this paper, we present iPAL's assessment framework and show how it guides the development of such performance assessments, exemplify these assessments with a concrete task, and provide preliminary evidence of its reliability and validity, which allows us to draw initial implications for further test design and development.
Abstract Background and aim There has been a surge of interest in the cardiovascular effects of vitamin D (25(OH)D), preventing the processes leading to vascular wall degeneration and coronary artery ...disease (CAD). Gender differences have been suggested for vitamin D status, with a higher rate of deficiency occurring especially in post-menopausal women, increasing the risk of bone fractures and osteoporosis. However, to date, few studies have evaluated the differences in 25(OH)D levels according to gender and their impact on the extent of CAD, which was therefore the aim of the present study. Methods and results In patients undergoing coronary angiography, fasting samples were collected for the assessment of 25(OH)D levels. Significant CAD was defined as at least one vessel stenosis >50%, while severe CAD was defined as left main and/or three-vessel disease. Of the 1811 patients included, 530 (29.3%) were females, who displayed older age ( p < 0.001), higher rate of renal failure ( p < 0.001), hypertension ( p = 0.05), treatment with angiotensin-receptor blockers ( p = 0.03) and diuretics ( p < 0.001), acute presentation ( p < 0.001), higher platelet count ( p < 0.001), glycosylated haemoglobin ( p = 0.02) and cholesterol ( p = 0.001), but an inverse relationship with smoking ( p < 0.001), previous cardiovascular events ( p < 0.001), treatment with statins and acetylsalicylic acid (ASA) ( p < 0.001), body mass index ( p = 0.002), haemoglobin ( p < 0.001), leucocytes ( p = 0.03) and triglycerides ( p < 0.001). Female gender was associated with lower vitamin D levels (14.5 ± 10.9 vs. 15.9 ± 9.5, p = 0.007) and independently associated with severe vitamin D deficiency (41.9% vs. 30.4%, p < 0.001; adjusted odds ratio (OR) (95% confidence interval (CI)) = 1.42 (1.08–1.87), p = 0.01). Lower tertiles of vitamin D were associated with an increased prevalence and severity of CAD in females (adjusted OR (95% CI = 1.26 (1.10–1.44), p = 0.001 for CAD; adjusted OR (95% CI) = 1.6 (1.39–1.87), p < 0.001 for severe CAD). In males, vitamin D status was independently related to the prevalence (adjusted OR (95% CI) = 1.28 (1.02–1.61), p = 0.03) of CAD, but not the extent of CAD (adjusted OR (95% CI) = 1.02 (0.86–1.2), p = 0.84). Conclusion Gender significantly affects vitamin D status. The lower 25(OH)D levels observed in females, as compared to males, play a more relevant role in conditioning the severity of CAD.
The clinical efficacy and safety of protein-based drugs such as monoclonal antibodies (mAbs) rely on the integrity of the protein higher order structure (HOS) during product development, ...manufacturing, storage, and patient administration. As mAb-based drugs are becoming more prevalent in the treatment of many illnesses, the need to establish metrics for quality attributes of mAb therapeutics through high-resolution techniques is also becoming evident. To this end, here we used a forced degradation method, time-dependent oxidation by hydrogen peroxide, on the model biotherapeutic NISTmAb and evaluated the effects on HOS with orthogonal analytical methods and a functional assay. To monitor the oxidation process, the experimental workflow involved incubation of NISTmAb with hydrogen peroxide in a benchtop nuclear magnetic resonance spectrometer (NMR) that followed the reaction kinetics, in real-time through the water proton transverse relaxation rate
(
H
O). Aliquots taken at defined time points were further analyzed by high-field 2D
H-
C methyl correlation fingerprint spectra in parallel with other analytical techniques, including thermal unfolding, size-exclusion chromatography, and surface plasmon resonance, to assess changes in stability, heterogeneity, and binding affinities. The complementary measurement outputs from the different techniques demonstrate the utility of combining NMR with other analytical tools to monitor oxidation kinetics and extract the resulting structural changes in mAbs that are functionally relevant, allowing rigorous assessment of HOS attributes relevant to the efficacy and safety of mAb-based drug products.
Protein therapeutics have numerous critical quality attributes (CQA) that must be evaluated to ensure safety and efficacy, including the requirement to adopt and retain the correct three-dimensional ...fold without forming unintended aggregates. Therefore, the ability to monitor protein higher order structure (HOS) can be valuable throughout the lifecycle of a protein therapeutic, from development to manufacture. 2D NMR has been introduced as a robust and precise tool to assess the HOS of a protein biotherapeutic. A common use case is to decide whether two groups of spectra are substantially different, as an indicator of difference in HOS. We demonstrate a quantitative use of principal component analysis (PCA) scores to perform this decision-making, and demonstrate the effect of acquisition and processing details on class separation using samples of NISTmAb monoclonal antibody Reference Material subjected to two different oxidative stress protocols. The work introduces an approach to computing similarity from PCA scores based upon the technique of histogram intersection, a method originally developed for retrieval of images from large databases. Results show that class separation can be robust with respect to random noise, reconstruction method, and analysis region selection. By contrast, details such as baseline distortion can have a pronounced effect, and so must be controlled carefully. Since the classification approach can be performed without the need to identify peaks, results suggest that it is possible to use even more efficient measurement strategies that do not produce spectra that can be analyzed visually, but nevertheless allow useful decision-making that is objective and automated.
Essentials
Dual antiplatelet therapy (DAPT) in elderly patients requires balancing bleedings and thrombosis.
Impact of age on high residual on‐treatment platelet reactivity (HRPR) on DAPT was ...studied.
A reduced effectiveness of adenosine diphosphate antagonists was observed over 70 years of age.
The occurrence of HRPR was increased among elderly patients with both clopidogrel and ticagrelor.
Summary
Background
The aim of the present study was to evaluate the impact of age on platelet function and the occurrence of high residual on‐treatment platelet reactivity (HRPR) in patients treated with dual antiplatelet therapy (DAPT) using acetylsalicilic acid (ASA) and clopidogrel or ticagrelor.
Methods
Patients treated with DAPT (ASA and clopidogrel or ticagrelor) were scheduled for platelet function assessment at 30–90 days post‐discharge. By whole blood impedance aggregometry, HRPR was considered for ASPI test values > 862 AU*min (for ASA) and adenosine diphosphate (ADP) test values > 417 AU*min (for ADP antagonists). Elderly patients were defined as those aged ≥ 70 years.
Results
Among 494 patients on DAPT, 224 (45.3%) were ≥ 70 years old. ADP‐mediated platelet aggregation increased with decades of age (279.3 ± 148.6 vs. 319.6 ± 171.1 vs. 347.3 ± 190.1 vs. 345.7 ± 169.2), whereas no difference was observed for ASA response.
A reduced effectiveness of ADP antagonists was observed among elderly patients; in fact, among the 117 patients displaying HRPR (23.7%), a higher prevalence was observed among patients over 70 years old (30.4% vs. 18.1%; adjusted odds ratio (OR) 95% confidence interval (CI) = 2.19 1.29–3.71). Similar results were obtained among the 266 clopidogrel‐treated patients (38.5% vs. 27.9%; adjusted OR 95% CI = 2.91 1.46–5.8) and in the 228 patients receiving ticagrelor (19.1% vs. 8.1%; adjusted OR 95% CI = 2.55 1.02–8.59).
Conclusion
In patients receiving dual antiplatelet therapy, advanced age is independently associated with a reduced effectiveness of ADP antagonists and a higher rate of HRPR with both clopidogrel and ticagrelor.
We present a theoretical study about the cation distribution and their effect on the physical properties of Ni-doped YIG system, based on a phenomenological model, the generalized Kapustinskii ...equation, and the single ion contribution to the saturation magnetization and cubic anisotropy constant. From the calculated reticular energy values (
U
ret
) it was possible to determine the more probable valence state configuration, corresponding with the coexistence of 3
+
and 2
+
oxidation states for Fe ions, the 2
+
oxidation state for Ni, and oxygen vacancies presence. In addition, from the occupation probability values, it was demonstrated that Ni
2+
and Fe
2+
cations prefer to occupy the octahedral sites, reflecting on the saturation magnetization (
M
s
) and cubic anisotropy constant (
K
1
) behaviors. The theoretical results of
M
s
and
K
1
were compared with the experimental values, showing a reasonable correspondence, considering the simplicity of the presented phenomenological model.
Lithium carbonate, a drug for the treatment of bipolar disorder, provides mood stability to mitigate recurrent episodes of mania and/or depression. Despite its long-term and widespread use, the ...mechanism by which lithium acts to elicit these psychological changes has remained unknown. Using nuclear magnetic resonance (NMR) methods, in this study we characterized the association of lithium with adenosine triphosphate (ATP) and identified a bimetallic (Mg·Li) ATP complex. Lithium’s affinity to form this complex was found to be relatively high (Kd ∼1.6 mM) compared with other monovalent cations and relevant, considering lithium dosing and physiological concentrations of Mg2+ and ATP. The ATP·Mg·Li complex reveals, for the first time, to the best of our knowledge, that lithium can associate with magnesium-bound phosphate sites and then act to modulate purine receptor activity in neuronal cells, suggesting a molecular mode for in vivo lithium action.
Toxoplasmosis affects one-third of the human population worldwide. Humans are accidental hosts and are infected after consumption of undercooked meat and water contaminated with Toxoplasma gondii ...cysts and oocysts, respectively. Neutrophils have been shown to participate in the control of T. gondii infection in mice through a variety of effector mechanisms, such as reactive oxygen species (ROS) and neutrophil extracellular trap (NET) formation. However, few studies have demonstrated the role of neutrophils in individuals naturally infected with T. gondii. In the current study, we evaluated the activation status of neutrophils in individuals with acute or chronic toxoplasmosis and determined the role of T. gondii-induced NET formation in the amplification of the innate and adaptive immune responses. We observed that neutrophils are highly activated during acute infection through increased expression of CD66b. Moreover, neutrophils from healthy donors (HDs) cocultured with tachyzoites produced ROS and formed NETs, with the latter being dependent on glycolysis, succinate dehydrogenase, gasdermin D, and neutrophil elastase. Furthermore, we observed elevated levels of the chemokines (CXC motif) CXCL8 and (CC motif) CCL4 ligands in plasma from patients with acute toxoplasmosis and production by neutrophils from HDs exposed to
. Finally, we showed that T. gondii-induced NETs activate neutrophils and promote the recruitment of autologous CD4
T cells and the production of interferon gamma (IFN-γ), tumor necrosis factor (TNF), interleukin 6 (IL-6), IL-17, and IL-10 by peripheral blood mononuclear cells. In conclusion, we demonstrated that T. gondii activates neutrophils and promotes the release of NETs, which amplify human innate and adaptive immune responses.
Approximately one-third of the human population is estimated to be chronically infected with the obligate intracellular parasite Toxoplasma gondii. Humans are accidental hosts that are infected with T. gondii after consumption of undercooked meat or contaminated water. Neutrophils have been shown to control T. gondii growth by different mechanisms, including neutrophil extracellular traps (NETs). In the current study, we observed that neutrophils are highly activated during acute toxoplasmosis. We also determined that T. gondii-induced NETs are dependent on the energetic profile of neutrophils as well as the production of ROS and gasdermin D (GSDMD) cleavage. In addition, we showed that T. gondii-induced NETs activate neutrophils, promote the recruitment of autologous CD4
T cells, and induce the production of cytokines by peripheral blood mononuclear cells, amplifying the innate and adaptive immune responses.
High impedance faults (HIFs) have been a major concern for protecting distribution systems and public safety hazards when involving downed conductors. The deployment of smarter grids brings new ...technologies for smart monitoring, automation, and protection of distribution networks. This paper presents a new method for a series of HIF detection and location in primary distribution feeders, using voltage unbalance measurements collected from smart meters (SMs) installed at low-voltage end-users. The methodology was tested in MATLAB and Simulink through steady-state simulations of a typical 13.8 kV distribution system, under load unbalance and different fault scenarios. Results show that the proposed method is robust and accurate for the detection of blown fuses and broken conductors, with or without ground faults, located either at the source or the load-side. The ease of implementation in SM design, formulation of parameters, and reliable simulation results show potential real-life applications.
Lentiviral (LV) vectors have emerged as powerful tools for treating genetic and acquired human diseases. As clinical studies and commercial demands have progressed, there has been a growing need for ...large amounts of purified LV vectors. To help meet this demand, we developed CRISPR library screening methods to identify genetic perturbations in human embryonic kidney 293 (HEK293) cells and their derivatives that may increase LV vector titers. Briefly, LV vector-based Human CRISPR Activation and Knockout libraries (Calabrese and Brunello) were used to modify HEK293 and HEK293T cells. These cell populations were then expanded, and integrated LV vector genomes were rescued by transfection. LV vectors were harvested, and the process of sequential transduction and rescue-transfection was iterated. Through this workflow, guide RNAs (gRNAs) that target genes that may suppress or enhance LV vector production were enriched and identified with Next-Generation Sequencing (NGS). Though more work is needed to test genes identified in this screen, we expect that perturbations of genes we identified here, such as
, which is an inhibitor of antiviral innate immunity may be introduced and multiplexed to yield cell lines with improved LV vector productivity.
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