Notes

Quantifying actions associated with day to day living impairment throughout Parkinson's ailment while using the Functional Activities List of questions.
The ability to synchronize a motor action to a rhythmic auditory stimulus is often considered an innate human skill. Protokylol However, some individuals lack the ability to synchronize speech to a perceived syllabic rate. Here, we describe a simple and fast protocol to classify a single native English speaker as being or not being a speech synchronizer. This protocol consists of four parts the pretest instructions and volume adjustment, the training procedure, the execution of the main task, and data analysis. For complete details on the use and execution of this protocol, please refer to Assaneo et al. (2019a).Identifying germline differentially methylated regions (DMRs) in outbred mammals remains a challenge because of difficulty in obtaining single-nucleotide polymorphisms (SNPs). To overcome this difficulty, we developed two computational approaches, TARSII and CARSII, which allow accurate prediction of germline DMRs from DNA methylomes independent of SNPs. Furthermore, we introduce an easy and quick way to validate the predicted germline DMRs with allelic DNA methylation using CGmapTools. Collectively, our strategy can greatly facilitate de novo identification of germline DMRs in outbred mammals. For complete details on the use and execution of this protocol, please refer to Chu et al. (2021).The present protocol describes the computational design of the SARS-CoV-2 receptor binding motif (RBD) to identify mutations that can potentially improve binding affinity for the human ACE2 (hACE2) receptor. We focus on four positions located at the interface with the hACE2 receptor in the RBDhACE2 complex. We conduct the design with a high-throughput computational protein design (CPD) program, Proteus, incorporating an adaptive Monte Carlo (MC) protocol that promotes the selection of sequences with good binding affinities. For complete details on the use and execution of this protocol, please refer to Polydorides and Archontis (2021).Two-electrode voltage clamp (TEVC) combined with the Xenopus laevis oocytes heterologous expression system is a powerful electrophysiological tool widely used to study the properties of many transmembrane proteins. Here, we describe a protocol using this combined approach to identify the ligands of odorant receptors that form ligand-gated ion channels. We detail the procedures for site-directed mutagenesis, oocyte microinjection, and TEVC recording. This protocol can also be used to identify the key residues and illustrate the structure-function relationships of these proteins. For complete details on the use and execution of this protocol, please refer to Cao et al. (2021).Classic approaches to characterizing cell cycle leverage chemicals or altered nucleotide pools, which could impact chromatin states at specific phases of the cell cycle. Such approaches could induce metabolic alterations and/or DNA damage, which could reshape protein recruitment and histone modifications. In this protocol, we describe ways to fix and sort cells across the cell cycle based on their DNA content. We further detail immunoprecipitation and library preparation, allowing analysis of the epigenome by chromatin immunoprecipitation sequencing (ChIP-seq) for small numbers of cells. For complete details on the use and execution of this protocol, please refer to Van Rechem et al. (2021).Quantifying differences in the amount of protein and mRNA caused by missense mutations in a gene of interest can be challenging, especially when using patient-derived primary cells, which are intrinsically variable. In this protocol, we describe how to culture patient-derived lymphoblast and fibroblast cell lines for later mRNA and protein quantification. We also describe the steps to examine variants of PUM1 in HEK293T cells, but the protocol can be applied to other proteins of interest. For complete details on the use and execution of this protocol, please refer to Gennarino et al. (2018).Genetic variants that affect neurological function will often produce changes visible at the level of gross morphology, either of the whole brain or of specific neuronal types. Here we describe how to perfuse and dissect the brain in preparation for Nissl staining. Then we outline steps for culturing mouse primary hippocampal neurons to evaluate dendritic arborization (Sholl analysis). For complete details on the use and execution of this protocol, please refer to Gennarino et al. (2018).The immunogenicity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) proteome is largely unknown. Here we describe a protocol for analyzing sera samples with SARS-CoV-2 proteome microarray. The proteins were expressed by either E. coli expression system or eukaryotic cell expression systems and obtained by affinity purification. The protocol includes microarray fabricating and sera profiling, which will be used to build an antibody response landscape for IgG and IgM. The protocol may help to facilitate a deeper understanding of immunity related to SARS-CoV-2. For complete details on the use and execution of this protocol, please refer to Li et al. (2021c).Laplace pressure is an important regulator of cell dynamics and behavior during cytokinesis. Here, we provide a protocol to measure Laplace pressure in cultured cells using a micropipette and describe the steps for imaging the actin cortex during cytokinesis. The quantification steps allow tracing dynamic change in Laplace pressure and displaying dynamic response of the actin cortex during cytokinesis in HeLa cells. This protocol can be applied to any cultured cell type during various stages of cell division. For complete details on the use and execution of this protocol, please refer to Wang et al. (2021).A ZnO/Ag/ZnO film with 2D periodic apertures can have a high visible transmittance and low emittances in atmospheric windows for visible-infrared compatible camouflage and a high transmittance in 5-8 μm for heat dissipation by radiation. This following protocol details the fabrication process of such a photonic structure based on photolithography technology. Also described are the related challenges and troubleshooting steps. For complete details on the use and execution of this protocol, please refer to Dang and Ye (2021).Recurrent seizures are a common feature in many neurologic disorders. Seizure examination may help with diagnosis, preclinical study, and development of treatment strategies. Here we detail protocols to prepare and implant electrodes, as well as to record and analyze seizure events in freely moving mice. SCA47 mice exhibit both preclinical seizures (i.e., epileptiform discharges of EEG) starting from ∼14 weeks of age and behavioral seizures (i.e., spontaneous behavioral seizures) starting from ∼22 weeks of age. For complete details on the use and execution of this protocol, please refer to Gennarino et al. (2018).Here, we present a streamlined protocol for assessing intracellular Mycobacterium tuberculosis (Mtb) loads in macrophages. This protocol describes the simultaneous assessment of macrophage viability using automated microscopy. Further, we detail the quantification of mycobacterial loads using a rapid, inexpensive, and accurate approach for mycobacterial DNA isolation from paraformaldehyde-fixed macrophages. Simultaneous assessment of the bacterial loads using internal standard and macrophage viability allows for precise quantification of the effects of perturbations on Mtb and host cells while accounting for technical artifacts. For complete details on the use and execution of this protocol, please refer to Chatterjee et al. (2021).In this study, modeling the COVID-19 pandemic via a novel fractional-order SIDARTHE (FO-SIDARTHE) differential system is presented. The purpose of this research seemed to be to show the consequences and relevance of the fractional-order (FO) COVID-19 SIDARTHE differential system, as well as FO required conditions underlying four control measures, called SI, SD, SA, and SR. The FO-SIDARTHE system incorporates eight phases of infection susceptible (S), infected (I), diagnosed (D), ailing (A), recognized (R), threatening (T), healed (H), and extinct (E). Our objective of all these investigations is to use fractional derivatives to increase the accuracy of the SIDARTHE system. A FO-SIDARTHE system has yet to be disclosed, nor has it yet been treated using the strength of stochastic solvers. Stochastic solvers based on the Levenberg-Marquardt backpropagation methodology (L-MB) and neural networks (NNs), specifically L-MBNNs, are being used to analyze a FO-SIDARTHE problem. Three cases having varied values under the same fractional order are being presented to resolve the FO-SIDARTHE system. The statistics employed to provide numerical solutions toward the FO-SIDARTHE system are classified as obeys 72% toward training, 18% in testing, and 10% for authorization. To establish the accuracy of such L-MBNNs utilizing Adams-Bashforth-Moulton, the numerical findings were compared with the reference solutions.In this study, we investigated whether playing an escape room game after explicit instruction (instruction-first group, N = 20) is more effective to learn about copyright and media law than playing the game before explicit instruction (problem-solving-first group, N = 21). This is an important question as escape room games are complex, problem-based learning environments that can overwhelm learners and thus hinder the acquisition of knowledge and skills. According to cognitive load theory, preparing learners with explicit instruction before problem-solving activities can overcome this problem leading to better learning. However, in the productive failure paradigm problem-solving before explicit instruction is seen as more effective, especially when it comes to the application of newly acquired knowledge to solve novel problems. Based on these two theories, we conducted an experiment and found that playing the developed digital escape room game after explicit instruction was more effective for knowledge retention and domain-specific self-efficacy with at the same time lower cognitive load. However, we found no differences regarding the application of knowledge as both groups scored equally high on transfer tasks. The result is discussed considering previous productive failure studies mostly conducted in the science and engineering domain. In sum, the instruction first approach proved to be effective for both knowledge acquisition and knowledge application, leading to higher domain-specific self-efficacy and lower cognitive load. Therefore, we conclude that the implementation of escape room games after instruction is an effective instructional approach and better suited to promote learning than playing escape room games before instruction.
The online version contains supplementary material available at 10.1186/s41039-022-00187-x.
The online version contains supplementary material available at 10.1186/s41039-022-00187-x.With the increasing prevalence in recent years, myopia has become an essential global health concern. In most instances, an increased axial length of the eye is the structural cause of nearsightedness. The scleral remodeling, primarily dependent on the scleral extracellular matrix (ECM) changes, is significantly linked to eye lengthening. Scleral remodeling plays a critical function in the incidence and progression of myopia. This mini-review will focus on recent research progress of scleral remodeling in the hope of providing new ideas for the prophylaxis and treatment of myopia.
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