Ultrasensitive Detection of Interleukin 6 by Using Silicon Nanowire Field-Effect Transistors.

Interleukin 6 (IL-6) has been regarded as a biomarker that can be applied as a predictor for the severity of COVID-19-infected patients. The IL-6 level also correlates well with respiratory dysfunction and mortality risk. In this work, three silanization approaches and two types of biorecognition elements were used on the silicon nanowire field-effect transistors (SiNW-FETs) to investigate and compare the sensing performance on the detection of IL-6. Experimental data revealed that the mixed-SAMs-modified silica surface could have superior surface morphology to APTES-modified and APS-modified silica surfaces. According to the data on detecting various concentrations of IL-6, the detection range of the aptamer-functionalized SiNW-FET was broader than that of the antibody-functionalized SiNW-FET. In addition, the lowest concentration of valid detection for the aptamer-functionalized SiNW-FET was 2.1 pg/mL, two orders of magnitude lower than the antibody-functionalized SiNW-FET. The detection range of the aptamer-functionalized SiNW-FET covered the concentration of IL-6, which could be used to predict fatal outcomes of COVID-19. The detection results in the buffer showed that the anti-IL-6 aptamer could produce better detection results on the SiNW-FETs, indicating its great opportunity in applications for sensing clinical samples.

Benefits of applying virtual reality in pelvic movement training through a Wii Fit: a randomized controlled trial.

BACKGROUND: Pelvic movement training has become compulsory for part of medical students. An increasing amount of research has focused on the influence of virtual reality (VR) on learning effectiveness. However, its application to pelvic floor muscles or pelvic movement training is still in its infancy. We compared the effectiveness of conventional pelvic movement training with or without VR-assisted pelvic movement training for student learning. METHODS: We recruited 44 university students (16 male and 28 female participants; average age = 19.7 ± 0.31 years) who had not previously received pelvic movement education or training. The participants were randomly assigned into traditional and experimental groups to acquire pelvic movements and relevant knowledge. The traditional group received conventional classes (about 15 min), whereas the experimental group received both conventional classes and VR-assisted teaching (additional VR session took approximately 25-45 min depending on the speed of movement of each participant). The participants were asked to control the trajectory of the centre of pressure on the Wii Fit balance board and build-in games to learn pelvic movements. We conducted evaluations before, immediately after, and 2 weeks after the experiment, based on the scores of written and practical examinations. The experimental group was also asked to complete a questionnaire during the posttest. RESULTS: We carried out two-way repeated measures ANOVA and discovered that the written examination scores indicated a significant Time × Group interaction (p=0.015). In each group, the written and practical examinations in the posttest and follow-up test exhibited significantly improved results compared with the baseline value (p <0.001, except for traditional group of written exam in follow up test vs. baseline p=0.001). The written examination in the follow-up test did not decline significantly compared with those in the posttest, but the practical examination in the follow-up test was decline significantly compared with those in the posttest (p=0.033). The experimental group had superior overall performance in the practical examinations than the traditional group (experimental group: mean = 76.27, 95% confidence level [CI] = 70.84-81.71; traditional group: mean = 64.21, 95% CI = 58.78-69.65). No significant difference in the written examination between two groups. The percentage for agreement ratio on the usefulness, ease of use, users' intention to continue using the VR-assisted teaching is high (95.5-100%). CONCLUSIONS: The results of this study suggested that conventional and conventional + VR teaching were both effective. However, the incorporation of VR stimulated learning motivation and facilitated precise performance of pelvic movements. It is recommended that pelvic floor muscles training could be supplemented with VR or games to increase students' motivation and understanding how to perform pelvic movements.

A Study on the Effects of Lateral-Wedge Insoles on Plantar-Pressure Pattern for Medial Knee Osteoarthritis Using the Wearable Sensing Insole.

Patients with knee osteoarthritis have a unique plantar-pressure pattern during walking, and lateral-wedge insoles are one of the treatment options. Participants were randomly assigned to either the lateral-wedge insole group or the ordinary insole group. The Visual Analog Scale (VAS), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), and plantar-pressure test scores were evaluated at the baseline and at 20 weeks. Plantar pressure data were collected using a pressure insole with 89 sensing locations. In the ordinary insole group, the function and total WOMAC scores decreased significantly (function score, 24.8 (baseline) to 16.5 (week 20); total score, 34.9 (baseline) to 24.6 (week 20)). During walking, the transverse width of the center of pressure as a percentage of foot width (%Trans) significantly increased in the ordinary insole group (baseline, 6.3%; week 20, 14.8%). In addition, the values of partial foot pressure as a percentage of body weight (%PFP) on the forefoot (baseline, 30.3%; week 20, 39.2%) and heel (baseline, 28.1%; week 20, 16.9%) also increased significantly in the ordinary insole group. Significant group-by-time interaction effects were observed for partial foot pressure per body weight in the forefoot (p = 0.031) and heel (p = 0.024). In the ordinary insole group, the plantar pressure on the heel significantly decreased (p = 0.011) and that on the forefoot significantly increased (p = 0.023). In contrast, plantar pressure remained stable in all regions in the lateral-wedge insole group. Thus, lateral-wedge insoles may protect against plantar pressure deterioration in patients with knee osteoarthritis.

In-Silico Selection of Aptamer Targeting SARS-CoV-2 Spike Protein.

Aptamers are single-stranded, short DNA or RNA oligonucleotides that can specifically bind to various target molecules. To diagnose the infected cases of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in time, numerous conventional methods are applied for viral detection via the amplification and quantification of DNA or antibodies specific to antigens on the virus. Herein, we generated a large number of mutated aptamer sequences, derived from a known sequence of receptor-binding domain (RBD)-1C aptamer, specific to the RBD of SARS-CoV-2 spike protein (S protein). Structural similarity, molecular docking, and molecular dynamics (MD) were utilized to screen aptamers and characterize the detailed interactions between the selected aptamers and the S protein. We identified two mutated aptamers, namely, RBD-1CM1 and RBD-1CM2, which presented better docking results against the S protein compared with the RBD-1C aptamer. Through the MD simulation, we further confirmed that the RBD-1CM1 aptamer can form the most stable complex with the S protein based on the number of hydrogen bonds formed between the two biomolecules. Based on the experimental data of quartz crystal microbalance (QCM), the RBD-1CM1 aptamer could produce larger signals in mass change and exhibit an improved binding affinity to the S protein. Therefore, the RBD-1CM1 aptamer, which was selected from 1431 mutants, was the best potential candidate for the detection of SARS-CoV-2. The RBD-1CM1 aptamer can be an alternative biological element for the development of SARS-CoV-2 diagnostic testing.

Comparing solution-gate and bottom-gate nanowire field-effect transistors on pH sensing with different salt concentrations and surface modifications.

Field-effect transistors (FETs) have been developed as pH sensors by using various device structures, fabrication technologies, and sensing film materials. Different transistor structures, like extended-gate (EG) FETs, floating-gate FET sensors, and dual-gate (DG) FETs, can enhance the sensor performance. In this article, we report the effects of using solution-gate and bottom-gate FET configurations on pH sensing and investigate the influence of different ionic concentrations of buffers in the measured signals. The surface charge of hafnium dioxide (HfO2) affected by the buffer pH, with/without the modification of polyethylene glycol (PEG) terminated with hydroxyl groups, and the location of applied gate voltage are vital factors to the sensor performance in pH sensing. Based on the results, the solution-gate FET exhibits good pH sensitivity even in the high ionic strength solutions of bis-tris propane (BTP), and these values of pH sensitivity are close to the Nernst limit (59.2 mV/pH). In general, silane-PEG-OH modification can reduce the deviations of measured signals in pH sensing. The performance of bottom-gate FET is inferior in the BTP buffers with high ionic solutions but suitable to be operated in low ionic concentrations, such as 0.1, 1, and 10 mM BTP buffers. The size of the ions was also studied and discussed. The solution-gate FET demonstrates excellent performance under high ionic strengths, meaning a more significant potential for detecting biological molecules under physiological conditions.

Paper/PMMA hybrid device with a microvalve-controlled design for exosome isolation and analysis.

This study proposes a paper/PMMA hybrid device designed to isolate exosomes and extract exosomal miRNA, followed by quantitative analysis. It aims to provide simplified and convenient sample preparation for potential point-of-care testing (POCT) processes. In contrast to previous work conducted by our research team, which focused on isolating exosomes and exosomal nucleic acids, this study introduces a novel approach by integrating paper and a PMMA mold with a microvalve controlled design. This innovative method enables the entire process to be performed on paper. The pressure on the paper could be adjusted by turning the screw upon the valve to change the pore size and permeability of the paper, which achieved the effect of controlling the flow rate of fluids. The paper was designed to have an immunoaffinity area for capturing exosomes and a sol-gel silica coating area for extracting miRNA. The paper-based ELISA (p-ELISA) exhibited a limit of detection and a limit of quantitation of 6 × 107 and 5.4 × 108 particles/mL, respectively, for exosome measurement. The reverse transcription quantitative polymerase chain reaction (RT-qPCR) revealed that the Ct (threshold cycle) value for quantifying the miR-21 in the miRNAs extracted by the proposed paper/PMMA hybrid device was comparable to the Ct value of the commercial extraction kit. The developed paper/PMMA hybrid device with a microvalve-controlled design should be incorporated into the POCT system to extract exosomal miRNAs.

Optimization of DNA-directed immobilization on mixed oligo(ethylene glycol) monolayers for immunodetection.

The development of protein chips has suffered from problems regarding long-term protein stability and activity. We present a protein sensor surface for immunodetection that is prepared by a DNA-directed protein immobilization method on a mixed self-assembled monolayer (SAM). By this approach, an immobilized single-stranded DNA (ssDNA) surface can be transferred/modified into a protein chip by flowing in ssDNA-conjugated protein when the protein chip measurement is needed. Therefore, the long-term stability of the protein chip will not be a problem for various applications. We tried various compositions for the SAM layer, the length of the ssDNA spacer, the end-point nucleotide composition, and the processes of ssDNA immobilization of the SAM for an optimized condition for shifting the DNA chip to a protein chip. The evaluations were made by using surface plasmon resonance. Our results indicated that a 50:1 ratio of oligo(ethylene glycol) (OEG)/COOH-terminated OEG and DNA sequences with 20mer are the best conditions found here for making a protein chip via a DNA-directed immobilization (DDI) method. The designed end-point nucleotide composition contains a few guanines or cytosines, and ssDNA immobilization of the SAM by dehybridizing immobilized double-stranded DNA (dsDNA) can improve the hybridization efficiency.

Improve sample preparation process for miRNA isolation from the culture cells by using silica fiber membrane.

In clinical applications of miRNAs, the purity and quality of the testing samples are very critical, especially the obtained tissue sample volume is limited. If the extracted miRNAs are contaminated or different in quality before analysis, it will increase the variance of the analysis result and make the medical information judgment incorrect and cannot be portable. Herein, we improved the commercially extraction kit by realizing the fundamental mechanism and hoped to serve finding optimal procedures for increasing the recovery of miRNAs extracted from cultured cells. In the adsorption process, the factors, like increasing the ethanol concentration or adding Ca2+, could influence the RNA adsorption were investigated. For the elution process, the effect caused by raising the elution temperature and raising the pH value of elution buffer was studied. Finally, the conditions for miRNA extraction are optimal modified by using a 65% (v/v) solution of ethanol in the adsorption process, and using TE buffer with the pH value of 8.0 and raising the temperature to 55 °C in the elution. According to the quantified results, the improved extraction kit can promote the recovery of endogenous miR-21 by about 6 times by using the optimal extraction conditions comparing with the miRNeasy Mini Kit.

Signal Enhancement of Silicon Nanowire Field-Effect Transistor Immunosensors by RNA Aptamer.

Silicon nanowire field-effect transistors (SiNW-FETs) have been demonstrated as a highly sensitive platform for label-free detection of a variety of biological and chemical entities. However, detecting signal from immunoassays by nano-FETs is severely hindered by the distribution of different charged groups of targeted entities, their binding orientation, and distances to the surface of the FET. Aptamers have been widely applied as a recognition element for plentiful biosensors because of small molecular sizes and moderate to high specific binding affinity with different types of molecules. In this study, we propose an effective approach to enhance the electrical responses of both direct (6×-histidine) and sandwich (amyloid ß 1-42) immunoassays in SiNW-FETs with R18, a highly negative charged RNA aptamer against rabbit immunoglobulin G (IgG). Empirical results presented that the immunosensors targeted with R18 expressed a significantly stabilized and amplified signal compared to the ones without this aptamer. The research outcome provides applicability of the highly negative charged aptamer as a bioamplifier for immunoassays by FETs.

Neutralized chimeric DNA probe for the improvement of GC-rich RNA detection specificity on the nanowire field-effect transistor.

Silicon nanowire (SiNW) field-effect transistors (FETs) is a powerful tool in genetic molecule analysis because of their high sensitivity, short detection time, and label-free detection. In nucleic acid detection, GC-rich nucleic acid sequences form self- and cross-dimers and stem-loop structures, which can easily obtain data containing signals from nonspecific DNA binding. The features of GC-rich nucleic acid sequences cause inaccuracies in nucleic acid detection and hinder the development of precision medicine. To improve the inaccurate detection results, we used phosphate-methylated (neutral) nucleotides to synthesize the neutralized chimeric DNA oligomer probe. The probe fragment originated from a primer for the detection of hepatitis C virus (HCV) genotype 3b, and single-mismatched and perfect-matched targets were designed for single nucleotide polymorphisms (SNP) detection on the SiNW FET device. Experimental results revealed that the HCV-3b chimeric neutralized DNA (nDNA) probe exhibited better performance for SNP discrimination in 10 mM bis-tris propane buffer at 25 °C than a regular DNA probe. The SNP discrimination of the nDNA probe could be further improved at 40 °C on the FET device. Consequently, the neutralized chimeric DNA probe could successfully distinguish SNP in the detection of GC-rich target sequences under optimal operating conditions on the SiNW FET device.

A Scoping Review of The Efficacy of Virtual Reality and Exergaming on Patients of Musculoskeletal System Disorder.

To assess the effects of virtual reality on patients with musculoskeletal disorders by means of a scoping review of randomized controlled trials (RCTs). The databases included PubMed, IEEE, and the MEDLINE database. Articles involving RCTs with higher than five points on the Physiotherapy Evidence Database (PEDro) scale were reviewed for suitability and inclusion. The methodological quality of the included RCT was evaluated using the PEDro scale. The three reviewers extracted relevant information from the included studies. Fourteen RCT articles were included. When compared with simple usual care or other forms of treatment, there was significant pain relief, increased functional capacity, reduced symptoms of the disorder, and increased joint angles for the virtual reality treatment of chronic musculoskeletal disorders. Furthermore, burn patients with acute pain were able to experience a significant therapeutic effect on pain relief. However, virtual reality treatment of patients with non-chronic pain such as total knee replacement, ankle sprains, as well as those who went through very short virtual reality treatments, did not show a significant difference in parameters, as compared with simple usual care and other forms of treatment. Current evidence supports VR treatment as having a significant effect on pain relief, increased joint mobility, or motor function of patients with chronic musculoskeletal disorders. VR seems quite effective in relieving the pain of patients with acute burns as well.

Synergetic improvements of sensitivity and specificity of nanowire field effect transistor gene chip by designing neutralized DNA as probe.

Neutral DNA analogs as probes for the detection of target oligomers on the biosensors based on the field-effect transistor (FET) configuration feature advantages in the enhancement of sensitivity and signal-to-noise ratio. Herein, we used phosphate-methylated nucleotides to synthesize two partially neutralized chimeric DNA products and a fully neutralized DNA sequence and adopted a regular DNA oligomer as probes on the polycrystalline silicon nanowire (NW) FET devices. The sequences of two neutralized chimeric DNAs close to the 5' end were alternately modified with the phosphate-methylated nucleotides, and all probes were immobilized via their 5' end on the NW surface. The non-specific-to-specific binding ratio indicated that the two 5'-end partially neutralized chimeric DNAs featured better performance than the regular and fully neutralized DNA oligomers. The partially neutralized probe design reduces the ionic strength needed for hybridization and increases the Debye length of detection, thus promoting the detection sensitivity of FET and achieving the limit of detection of 0.1 fM. By using an appropriate probe design, applying DNA oligomers with embedded phosphate-methylated nucleotides in the FET biosensors is a promising way for gene detection with high sensitivity and specificity.

The Combination of Computational and Biosensing Technologies for Selecting Aptamer against Prostate Specific Antigen.

Herein, we report a method of combining bioinformatics and biosensing technologies to select aptamers against prostate specific antigen (PSA). The main objective of this study is to select DNA aptamers with higher binding affinity for PSA by using the proposed method. Based on the five known sequences of PSA-binding aptamers, we adopted the functions of reproduction and crossover in the genetic algorithm to produce next-generation sequences for the computational and experimental analysis. RNAfold web server was utilized to analyze the secondary structures, and the 3-dimensional molecular models of aptamer sequences were generated by using RNAComposer web server. ZRANK scoring function was used to rerank the docking predictions from ZDOCK. The biosensors, the quartz crystal microbalance (QCM) and a surface plasmon resonance (SPR) instrument, were used to verify the binding ability of selected aptamer for PSA. By carrying out the simulations and experiments after two generations, we obtain one aptamer that can have the highest binding affinity with PSA, which generates almost 2-fold and 3-fold greater measured signals than the responses produced by the best known DNA sequence in the QCM and SPR experiments, respectively.

Kinetic analysis of beta-amyloid peptide aggregation induced by metal ions based on surface plasmon resonance biosensing.

Recent studies suggest that beta-amyloid (Abeta) aggregation and toxicity are facilitated by metal ions. This study aims to evaluate the kinetics of Abeta aggregation/dissociation in the presence of metal ions and to investigate the efficacy of a metal chelator to disrupt the metal ion-induced Abeta aggregates. Soluble Abeta(1-40) peptide was immobilized on a surface plasmon resonance biosensing surface and aggregation induced by contact with soluble Abeta with or without metal ions. Our study revealed that all the tested metal ions promoted Abeta aggregation but with different kinetics. Among them, Cu(II) ions had the highest association constant, and reached the maximum binding in 10 min. However, the Cu(II)-induced Abeta aggregates were unstable. Other ions attained the maximum Abeta binding at much longer times: 45 min for Ca(II), 60 min for Fe(II), Fe(III), and Zn(II) ions. The Abeta aggregates induced by Fe(III) ions had the greatest stability. The metal ion-induced Abeta(1-40) aggregates could be disrupted by the metal chelator, EDTA, suggesting a metal chelator may serve as a pharmacological agent to interfere with Abeta aggregation. Finally, this study demonstrates that the SPR biosensor can be an effective and efficient setup to investigate the mechanism of Abeta aggregation.

Computational selection of RNA aptamer against angiopoietin-2 and experimental evaluation.

Angiogenesis plays a decisive role in the growth and spread of cancer and angiopoietin-2 (Ang2) is in the spotlight of studies for its unique role in modulating angiogenesis. The aim of this study was to introduce a computational simulation approach to screen aptamers with high binding ability for Ang2. We carried out computational simulations of aptamer-protein interactions by using ZDOCK and ZRANK functions in Discovery Studio 3.5 starting from the available information of aptamers generated through the systematic evolution of ligands by exponential enrichment (SELEX) in the literature. From the best of three aptamers on the basis of ZRANK scores, 189 sequences with two-point mutations were created and simulated with Ang2. Then, we used a surface plasmon resonance (SPR) biosensor to test 3 mutant sequences of high ZRANK scores along with a high and a low affinity binding sequence as reported in the literature. We found a selected RNA aptamer has a higher binding affinity and SPR response than a reported sequence with the highest affinity. This is the first study of in silico selection of aptamers against Ang2 by using the ZRANK scoring function, which should help to increase the efficiency of selecting aptamers with high target-binding ability.

Failure of hydroxyapatite-coated acetabular cups. Ten-year follow-up of 85 Landos Atoll arthroplasties.

Between March 1990 and May 1991 we performed 85 primary total hip replacements in 74 patients using the Landos Atoll hydroxyapatite (HA)-coated cup and the Corail HA-coated stem. The patients were followed up for a mean of ten years. Of the 85 cups, 26 (31%) have already been revised and a further six are radiologically unstable and awaiting revision. Two femoral stems have been revised for infection without loosening. The retrieved acetabular cups were studied by SEM and image-processing techniques to quantify the amount of residual HA on the cup. This was correlated with the clinical variables and modes of failure. The residual HA (as a percentage of the surface) on the loose cups correlated negatively with the duration of implantation (r = -0.732, p < 0.001). Six cups were stable at revision and had more residual HA coating than those which were loose (p < 0.01). The rate of failure of the Landos Atoll HA-coated, smooth hemispherical cup with screw fixation is unacceptably high. Resorption of the HA coating is markedly increased in loose cups compared with stable cups. HA coating cannot substitute for stable mechanical fixation.

Geometric and morphological changes of the intervertebral disc under fatigue testing.

OBJECTIVE: To understand the injury mechanism of the intervertebral disc at different loading rates and to explore the anatomic and histological changes of intervertebral discs. DESIGN: Fresh porcine lumbar spines were used for fatigue testing to study the morphological changes of the intervertebral disc. BACKGROUND: Intervertebral disc problem is one of the most common causes that lead to low back pain. Slow repetitive loading was considered to be the critical factor of spine and disc injuries. METHODS: Twenty-four lumbar functional units were subjected to cyclic loading at three different loading rates. The geometric measurements and magnetic resonance image observations were conducted for the comprehension of morphological changes. The detail observation was taken through a stereomicroscope. RESULTS: There was no significance in geometric changes between different loading rates. For magnetic resonance imagings, morphological changes included the changes of nucleus pulposus shape, bulge of anterior and posterior longitudinal ligaments, and dehydration in annulus fibrosus. CONCLUSION: The morphological changes of intervertebral disc were revealed in certain kinds of lesions. The results imply that fatigue failure and degeneration or instability are strongly linked. The correlation of magnetic resonance imaging and anatomic observation showed a high correspondence in the comparison of shape and position of the nucleus pulpasus. RELEVANCE: The changes of geometric measurements and relationship between anatomic observation and magnetic resonance imaging finding had been analyzed. It could help in understanding the mechanism of triggering cause in the early stage of disc degeneration.

Improved DNA detection by utilizing electrically neutral DNA probe in field-effect transistor measurements as evidenced by surface plasmon resonance imaging.

Intensive efforts have been focused on the development of ultrasensitive DNA biosensors capable of quantitative gene expression analysis. Various neutralized nucleic acids have been demonstrated as alternative and attractive probe for the design of a DNA chip. However, the mechanism of the improvements has not been clearly revealed. In this investigation, we used a newly developed neutral ethylated DNA (E-DNA), a DNA analog with the "RO-P-O" backbone (wherein R could be methyl, ethyl, aryl, or alkyl group) obtained from synthetic procedures, and a silicon nanowire (SiNW) field-effect transistor (FET) to evaluate the difference in DNA detection performance while using E-DNA and DNA as probes. It is demonstrated that using the E-DNA probe in the FET measurement could have a significantly enhanced effect upon the detection sensitivity. Surface plasmon resonance imaging (SPRi) was used to evidence the mechanism of the improved detection sensitivity. SPRi analysis showed the amounts of probe immobilization on the sensor surface and the hybridization efficiency were both enhanced with the use of E-DNA. Consequently, neutral ethylated DNA probe hold a great promise for DNA sensing, especially in the electrical-based sensor.