Intracellular flow cytometry staining of antibody-secreting cells using phycoerythrin-conjugated antibodies: pitfalls and solutions

Antibody-secreting cells are terminally differentiated B cells that play a critical role in humoral immunity through immunoglobulin secretion along with possessing the potential to be long-lived. It is now appreciated that antibody-secreting cells regulate multiple aspects of biology through the secretion of various cytokines. In this regard, intracellular flow cytometry is a key tool used to assess the presence of intracellular proteins such as cytokines and transcription factors. Here, we showed that the use of phycoerythrin-containing antibody conjugates led to a false interpretation of antibody-secreting cell intracellular protein expression compared to other cell types. This was mainly due to the inappropriate retention of these antibodies specifically within antibody-secreting cells. Furthermore, we demonstrated how to reduce this retention which allowed for a more accurate comparison of intracellular protein expression between antibody-secreting cells and other cell types such as B lymphocytes. Using this methodology, our data revealed that spleen antibody-secreting cells expressed Toll-like receptor 7 as well as the pro-form of the inflammatory cytokine interleukin-1β.

Piezoelectric A15B16C17 Compounds and Their Nanocomposites for Energy Harvesting and Sensors: A Review

Interest in pyroelectrics and piezoelectrics has increased worldwide on account of their unique properties. Applications based on these phenomena include piezo- and pyroelectric nanogenerators, piezoelectric sensors, and piezocatalysis. One of the most interesting materials used in this growing field are A15B16C17 nanowires, an example of which is SbSI. The latter has an electromechanical coupling coefficient of 0.8, a piezoelectric module of 2000 pC/N, and a pyroelectric coefficient of 12 × 10−3 C/m2K. In this review, we examine the production and properties of these nanowires and their composites, such as PAN/SbSI and PVDF/SbSI. The generated electrical response from 11 different structures under various excitations, such as an impact or a pressure shock, are presented. It is shown, for example, that the PVDF/SbSI and PAN/SbSI composites have well-arranged nanowires, the orientation of which greatly affects the value of its output power. The power density for all the nanogenerators based upon A15B16C17 nanowires (and their composites) are recalculated by use of the same key equation. This enables an accurate comparison of the efficiency of all the configurations. The piezo- and photocatalytic properties of SbSI nanowires are also presented; their excellent ability is shown by the high reaction kinetic rate constant (7.6 min−1).

Seakeeping index as generalized indicator of ship seakeeping performance

Object and purpose of research. This paper discusses the possibility of a ship design process that would consider seakeeping performance to the greatest extent possible. The purpose of this study was to work out a numerical indicator, an index, reflecting all the seakeeping properties relevant for suitability of given ship to its intended operational conditions. Materials and methods. The study was based on the data about various operational parameters of the ship under investigation. These data were further synthesized so as to obtain the most comprehensive picture of ship seakeeping behaviour in different operational conditions. Main results. The study yielded the method and the algorithm for the “seakeeping index” as an average annual probability that seakeeping performance of given ship will be adequate to the conditions of given water area. The method sug-gested in this paper for a generalized comparison of seakeeping properties can handle whatever variety of target parameters and whatever seakeeping standards for any kind of ship intended to operate in given water area, and the result of this comparison is given in form of a single number that can be further used to improve seakeeping parameters of given ship, as well as to estimate possible time of its fully-featured operation in given conditions, including cost efficiency analysis. Conclusion. For more accurate comparison, it is recommended to analyse target parameters as functions of both ship speed and wave heading angle keeping in mind that the assumption introduced, i.e. that these curves as functions of wave heading angle are cosines, is not necessarily true. In other words, it is recommended to rely on more accurate data, experimental or analytical, so as to take into account the effect of apparent frequencies upon these curves.

Greig Cephalopolysyndactyly Contiguous Gene Syndrome: Case Report and Literature Review

Greig cephalopolysyndactyly syndrome (GCPS) is a rare genetic disorder (about 200 cases reported), characterized by macrocephaly, hypertelorism, and polysyndactyly. Most of the reported GCPS cases are the results of heterozygous loss of function mutations affecting the GLI3 gene (OMIM# 175700), while a small proportion of cases arise from large deletions on chromosome 7p14 encompassing the GLI3 gene. To our knowledge, only 6 patients have been reported to have a deletion with an exact size (given by genomic coordinates) and a gene content larger than 1 Mb involving the GLI3 gene. This report presents a patient with Greig cephalopolysyndactyly contiguous gene syndrome (GCP-CGS) diagnosed with a large, 18 Mb deletion on chromosome 7p14.2-p11.2. Similar cases are reviewed in the literature for a more accurate comparison between genotype and phenotype.

Trusted data spaces as a viable and sustainable solution for networks of population-based patient registries (Preprint)

BACKGROUND Population-based patient registries are entities that collect summary patient data from a well-defined population. Their main function is the monitoring and surveillance of a particular disease within their population catchment area, but they are also an important data source used in epidemiology. Comparing indicators across national boundaries brings considerable extra benefit to registries’ data, especially in regions where supranational initiatives are or could be coordinated to leverage good practices; this is particularly important for the European Union. Stricter data-protection laws however can unintentionally hamper the efforts of data harmonization to ensure the removal of statistical bias in the individual data sets, thereby compromising the integrated value of registries’ data. A new paradigm is required to ensure registries can operate in an environment that is not unnecessarily restrictive and allow accurate comparison of data for better ascertaining measures and practices most conducive to the public health of societies. OBJECTIVE To propose a solution towards a viable and sustainable model for the integration of registry data at supranational level. METHODS The pan-European organisational model of cancer registries, owing to its long and successful establishment, was taken as a good starting point from which to propose a sustainable, generic model for patient registries. Drawbacks to the model, particularly with respect to scalability and resourcing, were addressed in an adapted model. RESULTS An inter-registry organisational model based along the lines of the European Network of Cancer Registries was adapted to tackle the governance and resourcing aspects essential for a generic patient-registry model. The adapted model is a proposal for how patient registries can inter operate to ensure harmonisation and quality of data for accurate comparison at supranational level. CONCLUSIONS In view of the challenges relating to accurate and unbiased inter-comparison of population-based registry data across national boundaries for disease-surveillance purposes, a sustainable, generic patient-registry model is proposed. Integrating registry data is important for understanding progression and trends of the most prevalent diseases as well as for ascertaining effective control measures. The model promises a valuable data resource for epidemiological research, whilst providing a closely regulated environment for the processing of pseudonomised patient summary data on a broader scale than has hitherto been possible.

Resistive Switching Characteristics of ZnO-Based RRAM on Silicon Substrate

In this work, we conducted the following analysis of Ni/ZnO (20 nm)/n-type Si RRAM device with three different compliance currents (CCs). We compared I–V curves, including set, reset voltages, and resistance of LRS, HRS states for each CCs. For an accurate comparison of each case, statistical analysis is presented. In each case, the average value and the relative standard deviation (RSD) of resistance are calculated to analyze the characteristics of the distribution. The best variability is observed at higher CC (5 mA). In addition, we validated the non-volatile properties of the device using the retention data for each of the CCs. Based on this comparison, we proposed the most appropriate CC of the device operation. Also, a pulse was applied to measure the current waveform and demonstrate the regular operation of the device. Finally, the resistance of LRS and HRS states was measured by pulse. We statistically compared the measured pulse data with the DC data.

Enhancing Interrogation of Skeletal Muscle Samples for Informative Quantitative Data

Careful quantitative analysis of histological preparations of muscle samples is crucial to accurate investigation of myopathies in man and of interpretation of data from animals subjected to experimental or potentially therapeutic treatments. Protocols for measuring cell numbers are subject to problems arising from biases associated with preparative and analytical techniques. Prominent among these is the effect of polarized structure of skeletal muscle on sampling bias. It is also common in this tissue to collect data as ratios to convenient reference dominators, the fundamental bases of which are ill-defined, or unrecognized or not accurately assessable. Use of such ‘floating’ denominators raises a barrier to estimation of the absolute values that assume practical importance in medical research, where accurate comparison between different scenarios in different species is essential to the aim of translating preclinical research findings in animal models to clinical utility in Homo sapiens. This review identifies some of the underappreciated problems with current morphometric practice, some of which are exacerbated in skeletal muscle, and evaluates the extent of their intrusiveness into the of building an objective, accurate, picture of the structure of the muscle sample. It also contains recommendations for eliminating or at least minimizing these problems. Principal among these, would be the use of stereological procedures to avoid the substantial counting biases arising from inter-procedure differences in object size and section thickness. Attention is also drawn to the distortions of interpretation arising from use of undefined or inappropriate denominators.

Comparison of Electrical Characterization of AlGaN/GaN HEMT with Multi-finger Gate

This paper presents an exhaustive TCAD based comparison of the multi-gate and T-gate AlGaN/GaN HEMT. This paper simulates the DC and RF characteristics of the device and makes an accurate comparison. The important feature of the device such as threshold voltage, drain current output characteristics, transconductance, cut-off frequency, and maximum oscillation frequency were obtained. It is concluded that the shape optimization of the HEMT with multi-finger gates and the advantages over traditional T-gate devices. The device of two-finger gate with 50nm spacing has the best output characteristics, and its maximum saturation current is about 110% the size of the device of four-finger gate with 200nm at VGS=0V. And the gm and the gain of the device with 50nm spacing two-finger gate is 76mS/mm larger than the HEMT of three-finger gate with 200nm. In addition, we also conducted a simulation in the case of changing only refers to finger-gate length and cap-gate length. And it is concluded that the two-finger gate HEMT with 250nm finger-gate length and 2.0µm cap-gate length has the best output characteristics, which output current is 0.159 A/µm at VGS=-1.5V. The results show that AlGaN/GaN HEMT with multi-finger gate have great potential for high power and high frequency applications electronic devices.

The Need for Community Standards to Enable Accurate Comparison of Glycoproteomics Algorithm Performance

Protein glycosylation that mediates interactions among viral proteins, host receptors, and immune molecules is an important consideration for predicting viral antigenicity. Viral spike proteins, the proteins responsible for host cell invasion, are especially important to be examined. However, there is a lack of consensus within the field of glycoproteomics regarding identification strategy and false discovery rate (FDR) calculation that impedes our examinations. As a case study in the overlap between software, here as a case study, we examine recently published SARS-CoV-2 glycoprotein datasets with four glycoproteomics identification software with their recommended protocols: GlycReSoft, Byonic, pGlyco2, and MSFragger-Glyco. These software use different Target-Decoy Analysis (TDA) forms to estimate FDR and have different database-oriented search methods with varying degrees of quantification capabilities. Instead of an ideal overlap between software, we observed different sets of identifications with the intersection. When clustering by glycopeptide identifications, we see higher degrees of relatedness within software than within glycosites. Taking the consensus between results yields a conservative and non-informative conclusion as we lose identifications in the desire for caution; these non-consensus identifications are often lower abundance and, therefore, more susceptible to nuanced changes. We conclude that present glycoproteomics softwares are not directly comparable, and that methods are needed to assess their overall results and FDR estimation performance. Once such tools are developed, it will be possible to improve FDR methods and quantify complex glycoproteomes with acceptable confidence, rather than potentially misleading broad strokes.

SING AND MOVE A SONG FOR LEARNING VOCABULARY IN KINDERGARTEN

English language skills are expected to help children in the future to be able to interact globally and make it easier for them to establish relationships widely. One of the concerns in learning English is to build vocabulary in children through fun activities so that children do not feel that learning English is too boring and difficult. One of the methods used to introduce English to children is to use fun methods such as movements and songs which are usually done as a routine in class. This study aims to determine how the level of students' abilities and students' perceptions in improving English vocabulary after being given treatment using motion and song. This study used quantitative methods through pre-experimental design that utilizes pre-test and post-test to a group of students to determine their progress before and after being given treatment. Using the test would make an appropriate and accurate comparison to find out the improvement that occurs to students. The data collection technique was carried out by classroom observation at one of the Kindergartens that had compulsory English extracurricular. With the results in the post-test, the findings of this study were that early childhood children could have significant results after being given treatment. There were even children who can answer almost all the vocabulary in the ten designated objects. In addition, based on the responses of students who took part in the lesson they reveled this method with a thumbs up sign and a smile shown to the teacher. So it can be concluded that the motion and song method can be suitable methods for early childhood to introduce English.