Immunogenicity, reactogenicity, and safety of two-dose adjuvanted herpes zoster subunit vaccine in patients with systemic lupus erythematosus in South Korea: a single-centre, randomised, double-blind, placebo-controlled trial.

BACKGROUND: The adjuvanted herpes zoster subunit vaccine has shown good efficacy and safety in the general population. However, its effectiveness has not been comprehensively assessed in patients with systemic lupus erythematosus (SLE). This study aimed to evaluate the immunogenicity and safety of the adjuvanted herpes zoster subunit vaccine in patients with SLE. METHODS: This single-centre, randomised, double-blind, placebo-controlled, trial was done at the rheumatology outpatient clinic at Seoul National University Hospital, South Korea. Patients (aged ≥19 years) with clinically stable SLE and previous exposure (≥4 weeks) to immunosuppressive drugs were randomly assigned (4:1) via a central interactive web response system to receive herpes zoster subunit vaccine or placebo (0·5 mL intramuscular injection) at weeks 0 and 8. Investigators and participants were masked to intervention and group assignment. Anti-glycoprotein E antibody titres and glycoprotein E-specific cell-mediated vaccine responses were evaluated at baseline and at week 8 after the first dose, and at week 4, week 26, and week 52 after the second dose using enzyme-linked immunosorbent assay and flow cytometry, respectively. Reactogenicity, SLE disease activity, including Systemic Lupus Erythematosus Disease Activity Index 2000 and British Isles Lupus Assessment Group-flare rate, were examined. The primary outcome was the proportion of patients with a positive humoral vaccine response 4 weeks after the second dose. The primary and safety analyses were done in a modified intention-to-treat population. This study is registered with ClinicalTrials.gov, NCT06001606. FINDINGS: Between June 14, and July 19, 2023, 65 patients with SLE were enrolled, of whom 52 were randomly assigned to the herpes zoster subunit vaccine and 13 to placebo. 49 patients in the vaccine group and 11 patients in the placebo group were included in the modified intention-to-treat population. 56 (93%) of 60 patients were women and four (7%) were men. Mean age was 48·7 years (SD 11·4). The proportion of participants with a humoral vaccine response at 4 weeks after the second dose was significantly higher in the vaccine group (48 [98%] of 49 participants) than the placebo group (none [0%] of 11 patients; p<0·0001). More patients in the vaccine group than placebo group reported injection site reactions (42 patients vs two patients), fever (ten vs none), and fatigue (26 vs two). There were no differences in Systemic Lupus Erythematosus Disease Activity Index 2000 and British Isles Lupus Assessment Group-flare rates between the groups. There were no treatment-related deaths. INTERPRETATION: The herpes zoster subunit vaccine induces humoral and cellular immunity against herpes zoster with a good safety profile in patients with SLE. A larger study is warranted to assess the efficacy of vaccines to prevent herpes zoster in patients with SLE. FUNDING: Ministry of Science and ICT, The Government of the Republic of Korea.

Soluble expression of recombinant coagulation factor IX protein using Escherichia coli.

Hemophilia B is a congenital bleeding disorder caused by factor IX (FIX) deficiency. Generation of recombinant FIX (rFIX) is required for detecting a Hemophilia B indicator, anti-FIX antibody. In this study, we described a method for producing recombinant FIX (rFIX) using Escherichia coli. We constructed a FIX-expressing plasmid without a fusion tag protein-encoding gene and produced rFIX as a soluble form within five days. Dose-dependent curve was obtained from ELISA using anti-FIX antibody, indicating that the rFIX can be used as an antigen to detect anti-FIX antibody with high affinity and sensitivity.

Biosignal-integrated robotic systems with emerging trends in visual interfaces: A systematic review.

Human-machine interfaces (HMI) are currently a trendy and rapidly expanding area of research. Interestingly, the human user does not readily observe the interface between humans and machines. Instead, interactions between the machine and electrical signals from the user's body are obscured by complex control algorithms. The result is effectively a one-way street, wherein data is only transmitted from human to machine. Thus, a gap remains in the literature: how can information be effectively conveyed to the user to enable mutual understanding between humans and machines? Here, this paper reviews recent advancements in biosignal-integrated wearable robotics, with a particular emphasis on "visualization"-the presentation of relevant data, statistics, and visual feedback to the user. This review article covers various signals of interest, such as electroencephalograms and electromyograms, and explores novel sensor architectures and key materials. Recent developments in wearable robotics are examined from control and mechanical design perspectives. Additionally, we discuss current visualization methods and outline the field's future direction. While much of the HMI field focuses on biomedical and healthcare applications, such as rehabilitation of spinal cord injury and stroke patients, this paper also covers less common applications in manufacturing, defense, and other domains.

Evaluation of PDL1 positive cancer cell-specific binding activity of recombinant anti-PDL1 scFv.

Programmed cell death-ligand 1 (PDL1) is a transmembrane protein that is characterized as an immune regulatory molecule. We recently developed a recombinant single-chain fragment of variable domain (scFv) against PDL1, which showed high binding efficiency to purified recombinant PDL1 protein. However, at that time, proof-of-concept data for the effect of scFv using PDL1-expressing cells was lacking. In this study, we conducted two kinds of cell-based immunoassays, western blotting and enzyme-linked immunosorbent assay, using anti-PDL1 scFv. The results indicate that scFv can selectively and sensitively detect PDL1 from PDL1 positive human cancer cell lines. Our findings suggest that scFv could be used as a potential PDL1 inhibitor agent and probe for cell-based immunoassays to detect PDL1.

Fluorogenic enzyme-linked immunosorbent assay with a dual color variation.

The development of accurate and high-throughput biomarker detection tools is crucial for the diagnosis, monitoring, and treatment of various diseases. In this study, a sensitive fluorogenic enzyme-linked immunosorbent assay (FELISA) using Amplex Red or QuantaBlu fluorescent substrate was developed for the detection of tumor necrosis factor alpha and programmed cell death-ligand 1. The limit of detection of FELISA was in the nanogram order and multiple samples were conveniently assayed within 20 h using FELISA, demonstrating its applicability as a powerful immunoassay tool. FELISA can be widely used for rapid and accurate TNFα and PDL1 detection and applied to various fluorogenic immunoassays against other antigens of interest.

Development of fluorescence-linked immunosorbent assay for rapid detection of Staphylococcus aureus.

Staphylococcus aureus is a major pathogen that causes infections and life-threatening diseases. Although antibiotics, such as methicillin, have been used, methicillin-resistant S. aureus (MRSA) causes high morbidity and mortality rates, and conventional detection methods are difficult to be used because of time-consuming process. To control the spread of S. aureus, a development of a rapid and simple detection method is required. In this study, we generated a fluorescent anti-S. aureus antibody, and established a novel fluorescence-linked immunosorbent assay (FLISA)-based S. aureus detection method. The method showed high sensitivity and low limit of detection toward MRSA detection. The assay time for FLISA was 5 h, which was faster than that of conventional enzyme-linked immunosorbent assay (ELISA) or rapid ELISA. Moreover, the FLISA-based detection method was applied to diagnose clinically isolated MRSA samples that required only 5.3 h of preincubation. The FLISA method developed in this study can be widely applied as a useful tool for convenient S. aureus detection. KEY POINTS: • A fluorescence-linked immunosorbent assay-based S. aureus detection method • Simultaneous quantification of a maximum of 96 samples within 5 h • Application of the novel system to diagnosis clinical isolates.

Physiological Indicators of Fluency and Engagement during Sequential and Simultaneous Modes of Human-Robot Collaboration.

Occupational ApplicationsAn understanding of fluency in human-robot teaming from a physiological standpoint is still incomplete. In our experimental study involving 24 participants, we designed a scenario for shared-space human-robot collaboration (HRC) for a material sorting task. When compared to a sequential mode of interaction, the simultaneous mode resulted in significantly higher perceptions of fluency and engagement, primarily by reducing human idle time. These observations were complemented by significant changes in physiological responses, such as ECG entropy and low frequency power. These responses could predict fluency and engagement with accuracies of 90 and 97%, respectively. Notably, the perception of fluency and preferred mode of interaction were influenced by individual preferences. Hence, it is crucial to consider both physiological responses and user preferences when designing HRC systems, to ensure a positive experience with the robot teammate and to foster engagement in long-term teamwork. Furthermore, these signals can be obtained using a single robust, low-cost, and comfortable sensor.

Background In the current industry, a key enabler of flexible manufacturing is human-robot collaboration (HRC), a scenario wherein a human and a robot interact and work together in a shared space to accomplish a common task. In HRC, the timing and coordination between the human and robot are crucial factors that impact the fluency, efficacy, and acceptance of human-robot teams.Purpose Experimental research on the physiological indicators of fluency in human-robot collaborative tasks in a shared workspace is still in its infancy. We posit that by relating the mental perceptions of fluency to features from physiological signals, we could bring more clarity to the complex mapping between subjective and objective measures of fluency.Methods Twenty-four participants (12 males and 12 females), with mean (SD) age = 25.7 (2.9) years, completed an experimental study. We investigated the effects of interaction mode (sequential, simultaneous) and level of human involvement (low, medium, high) on perceived fluency, engagement, performance, and physiological response (heart rate variability = HRV) in a collaborative item sorting task.Results The simultaneous mode of interaction and a higher level of human involvement led to higher ratings for fluency and engagement, along with ECG changes, specifically an 11.6% increase in low-frequency power and a 3% reduction in information entropy. Using machine learning, these HRV features could predict perceived fluency and engagement with 90 and 97% accuracy, respectively.Conclusion Our results indicate that a human operator's perceived fluency in human-robot collaborative tasks can be measured using HRV metrics. Our findings expand the current fluency framework from a physiological perspective and offer additional objective measures derived from HRV, which could be practically applied to improve the design and optimization of HRC systems.

Quenchbodies That Enable One-Pot Detection of Antigens: A Structural Perspective.

Quenchbody (Q-body) is a unique, reagentless, fluorescent antibody whose fluorescent intensity increases in an antigen-concentration-dependent manner. Q-body-based homogeneous immunoassay is superior to conventional immunoassays as it does not require multiple immobilization, reaction, and washing steps. In fact, simply mixing the Q-body and the sample containing the antigen enables the detection of the target antigen. To date, various Q-bodies have been developed to detect biomarkers of interest, including haptens, peptides, proteins, and cells. This review sought to describe the principle of Q-body-based immunoassay and the use of Q-body for various immunoassays. In particular, the Q-bodies were classified from a structural perspective to provide useful information for designing Q-bodies with an appropriate objective.

Generation of a Recombinant scFv against Deoxycholic Acid and Its Conversion to a Quenchbody for One-Step Immunoassay.

Development of a rapid detection method for deoxycholic acid (DCA) is crucial for its diagnosis in the early stages of inflammation and cancer. In this study, we expressed a soluble recombinant anti-DCA single-chain variable fragment (scFv) in Escherichia coli. To convert scFv into a Quenchbody (Q-body), we labeled scFv using commercially available maleimide-linked fluorophores. The TAMRA-C5-maleimide-conjugated Q-body showed the highest response within a few minutes of DCA addition, indicating its applicability as a wash-free immunoassay probe for onsite DCA detection.

Production and engineering of nanobody-based quenchbody sensors for detecting recombinant human growth hormone and its isoforms.

Due to athletes' misuse of recombinant human growth hormone (rhGH) for performance improvement, the World Anti-Doping Agency has designated rhGH as a prohibited substance. This study focuses on the development and improvement of a simple and fast rhGH detection method using a fluorescence-incorporated antibody sensor "Quenchbody (Q-body)" that activates upon antigen binding. Camelid-derived nanobodies were used to produce stable Q-bodies that withstand high temperatures and pH levels. Notably, pituitary human growth hormone (phGH) comprises two major isoforms, namely 22 and 20 kDa GH, which exist in a specific ratio, and the rhGH variant shares the same sequence as the 22 kDa GH isoform. Therefore, we aimed to discriminate rhGH abuse by analyzing its specific isoform ratio. Two nanobodies, NbPit (recognizing phGH) and NbRec (preferentially recognizing 22 kDa rhGH), were used to develop the Q-bodies. Nanobody production in Escherichia coli involved the utilization of a vector containing 6xHis-tag, and Q-bodies were obtained using a maleimide-thiol reaction between the N-terminal of the cysteine tag and a fluorescent dye. The addition of tryptophan residue through antibody engineering resulted in increased fluorescence intensity (FI) (from 2.58-fold to 3.04-fold). The limit of detection (LOD) was determined using a fluorescence response, with TAMRA-labeled NbRec successfully detecting 6.38 ng/ml of 22 kDa rhGH while unable to detect 20 kDa GH. However, ATTO520-labeled NbPit detected 7.00 ng/ml of 20 kDa GH and 2.20 ng/ml 22 kDa rhGH. Q-bodies successfully detected changes in the GH concentration ratio from 10 to 40 ng/ml in human serum within 10 min without requiring specialized equipment and kits. Overall, these findings have potential applications in the field of anti-doping measures and can contribute to improved monitoring and enforcement of rhGH misuse, ultimately enhancing fairness and integrity in competitive sports.

Predicting the metabolic cost of exoskeleton-assisted squatting using foot pressure features and machine learning.

Introduction: Recent studies found that wearable exoskeletons can reduce physical effort and fatigue during squatting. In particular, subject-specific assistance helped to significantly reduce physical effort, shown by reduced metabolic cost, using human-in-the-loop optimization of the exoskeleton parameters. However, measuring metabolic cost using respiratory data has limitations, such as long estimation times, presence of noise, and user discomfort. A recent study suggests that foot contact forces can address those challenges and be used as an alternative metric to the metabolic cost to personalize wearable robot assistance during walking. Methods: In this study, we propose that foot center of pressure (CoP) features can be used to estimate the metabolic cost of squatting using a machine learning method. Five subjects' foot pressure and metabolic cost data were collected as they performed squats with an ankle exoskeleton at different assistance conditions in our prior study. In this study, we extracted statistical features from the CoP squat trajectories and fed them as input to a random forest model, with the metabolic cost as the output. Results: The model predicted the metabolic cost with a mean error of 0.55 W/kg on unseen test data, with a high correlation (r = 0.89, p < 0.01) between the true and predicted cost. The features of the CoP trajectory in the medial-lateral direction of the foot (xCoP), which relate to ankle eversion-inversion, were found to be important and highly correlated with metabolic cost. Conclusion: Our findings indicate that increased ankle eversion (outward roll of the ankle), which reflects a suboptimal squatting strategy, results in higher metabolic cost. Higher ankle eversion has been linked with the etiology of chronic lower limb injuries. Hence, a CoP-based cost function in human-in-the-loop optimization could offer several advantages, such as reduced estimation time, injury risk mitigation, and better user comfort.

Bacterial Production of Recombinant Coagulation Factor VIII Domains.

Factor VIII (F8) is a blood coagulation protein prearranged in six domains, and its deficiency causes hemophilia A. To fashion functional F8 therapeutics, development of a recombinant F8 (rF8) domain is essential not only for F8 substitution, but also to decipher the F8-related mechanisms. In this study, we generated Glutathione S-transferase (GST)-conjugated recombinant A2 and A3 domains of F8 using Escherichia coli. The high growth rate and economically advantageous protein production system in terms of inexpensive reagents and materials in E. coli cells facilitated the completion of entire process from protein expression to purification in 3-4 days with low production cost. Subsequent assessment of these purified proteins using enzyme-linked immunosorbent assay (ELISA) and antibodies against F8 revealed enhanced detection of rF8-A2 or rF8-A3 in a concentration dependent manner, indicating the presence of the antibody-binding epitopes in these proteins. Furthermore, these proteins are suitable for generating novel antibodies against the F8 domain and F8 domain-capturing affinity columns by enabling their conjugation to GST-capturing beads. Additionally, the recombinant F8 domains produced herein can be used for various studies, which include investigating the explicit roles of the F8 domain in the coagulation process, with domain-specific binding partners, and antibodies.

Generation of Q-bead against bone Gla protein with simplified preparation steps.

Quenchbody (Q-body)-based immunoassays enable the detection of antigen within a few minutes with high sensitivity and specificity, thereby revealing their applicability as biosensors for quantifying several biomolecules of interest; however, while producing a Q-body, it is necessary to eliminate the unconjugated dye after labeling to separate the Q-body from the capturing bead and to change the buffer using ultrafiltration, which is time-consuming and leads to yield reduction. In this study, we generated a recombinant single chain variable fragment against bone Gla protein as a model antibody. We labeled the antibody with a dye to generate a Q-body and subsequently added affinity beads to the Q-body mixture. After washing, we directly added antigen without extracting the Q-body from the bead and then measured the fluorescence intensity. As a result, the antigen-dependent fluorescence response was obtained from "Q-bead", which was almost the same as that of the Q-body generated according to the conventional method. The Q-bead was generated within only 2.5 h, thus requiring an hour and two steps less than those required for the generation of the traditional Q-body. No expensive Flag peptide was required to recover the total antibody from beads. Moreover, the ultra-filtration step was eliminated in this bead-based method, leading to improved convenience and cost- and time-saving attributes. The Q-bead-based assay can be used as a standard protocol for simple and rapid analysis of antibody-based molecular detection.

Synthesis and biological evaluation of N-(3-fluorobenzyl)-4-(1-(methyl-d3)-1H-indazol-5-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-amine as a novel, potent ALK5 receptor inhibitor.

Specific inhibition of ALK5 provides a novel method for controlling the development of cancers and fibrotic diseases. In this work, a novel series of N-(3-fluorobenzyl)-4-(1-(methyl-d3)-1H-indazol-5-yl)-5-(6-methylpyridin-2-yl)-1H-imidazol-2-amine (11), a potential clinical candidate, was synthesized by strategic incorporation of deuterium at potential metabolic soft spots and identified as ALK5 inhibitors. This compound has a low potential for CYP-mediated drug-drug interactions as a CYP450 inhibitor (IC50 = >10 µM) and showed potent inhibitory effects in cellular assay (IC50 = 3.5 ± 0.4 nM). The pharmacokinetic evaluation of 11 in mice demonstrated moderate clearance (29.0 mL/min/kg) and also revealed high oral bioavailability in mice (F = 67.6%).

Generation of a novel monoclonal antibody against inflammatory biomarker S100A8 using hybridoma technology.

OBJECTIVES: S100A8 is highly expressed in several inflammatory and oncological conditions. To address the current lack of a reliable and sensitive detection method for S100A8, we generated a monoclonal antibody with a high binding affinity to human S100A8 to enable early disease diagnosis. RESULTS: A soluble recombinant S100A8 protein with a high yield and purity was produced using Escherichia coli. Next, mice were immunized with recombinant S100A8 to obtain anti-human S100A8 monoclonal antibodies using hybridoma technology. Lastly, the high binding activity of the antibody was confirmed and its sequence was identified. CONCLUSIONS: This method, including the production of antigens and antibodies, will be useful for the generation of hybridoma cell lines that produce anti-S100A8 monoclonal antibodies. Moreover, the sequence information of the antibody can be used to develop a recombinant antibody for use in various research and clinical applications.

Generation of a novel antibody against BxPrx, a diagnostic marker of pine wilt disease.

BACKGROUND: Bursaphelenchus xylophilus is a pathogenic nematode that causes pine wilt disease (PWD). To prevent the rapid spread of this pathogen, developing a method for rapid and accurate detection of B. xylophilus is required. METHODS AND RESULTS: In this study, we produced a B. xylophilus peroxiredoxin (BxPrx), which is a protein that is overexpressed in B. xylophilus. Using recombinant BxPrx as an antigen, we generated and selected a novel antibody that binds to BxPrx via phage display and biopanning. We subcloned the anti-BxPrx single-chain variable fragment-encoding phagemid DNA to mammalian expression vector. We transfected the plasmid into mammalian cells and produced a highly sensitive recombinant antibody that enabled nanogram order detection of BxPrx. CONCLUSION: The sequence of anti-BxPrx antibody as well as the rapid immunoassay system described here can be applied for rapid and accurate diagnosis of PWD.

Effects of an intelligent virtual assistant on office task performance and workload in a noisy environment.

This study examines the effects of noise and the use of an Intelligent Virtual Assistant (IVA) on the task performance and workload of office workers. Data were collected from forty-eight adults across varied office task scenarios (i.e., sending an email, setting up a timer/reminder, and searching for a phone number/address) and noise types (i.e., silence, non-verbal noise, and verbal noise). The baseline for this study is measured without the use of an IVA. Significant differences in performance and workload were found on both objective and subjective measures. In particular, verbal noise emerged as the primary factor affecting performance using an IVA. Task performance was dependent on the task scenario and noise type. Subjective ratings found that participants preferred to use IVA for less complex tasks. Future work can focus more on the effects of tasks, demographics, and learning curves. Furthermore, this work can help guide IVA system designers by highlighting factors affecting performance.

Muscle coordination and recruitment during squat assistance using a robotic ankle-foot exoskeleton.

Squatting is an intensive activity routinely performed in the workplace to lift and lower loads. The effort to perform a squat can decrease using an exoskeleton that considers individual worker's differences and assists them with a customized solution, namely, personalized assistance. Designing such an exoskeleton could be improved by understanding how the user's muscle activity changes when assistance is provided. This study investigated the change in the muscle recruitment and activation pattern when personalized assistance was provided. The personalized assistance was provided by an ankle-foot exoskeleton during squatting and we compared its effect with that of the no-device and unpowered exoskeleton conditions using previously collected data. We identified four main muscle recruitment strategies across ten participants. One of the strategies mainly used quadriceps muscles, and the activation level corresponding to the strategy was reduced under exoskeleton assistance compared to the no-device and unpowered conditions. These quadriceps dominant synergy and rectus femoris activations showed reasonable correlations (r = 0.65, 0.59) to the metabolic cost of squatting. These results indicate that the assistance helped reduce quadriceps activation, and thus, the metabolic cost of squatting. These outcomes suggest that the muscle recruitment and activation patterns could be used to design an exoskeleton and training methods.

Tiered human health risk assessment of antibacterial quaternary ammonium compounds (QACs) in dishwashing detergents.

Quaternary ammonium compounds (QACs) are widely used in consumer products because of their unique antibacterial properties, and dishwashing detergents are a major source of exposure through oral, inhalation, and dermal routes. The three classes of QACs, including benzalkonium chloride (BAC), n-alkyldimethylethylbenzylammonium chloride (ADEBAC), and di-n-alkyldimethylammonium chloride (DDAC), in spray and non-spray types of dishwashing detergents were quantified by high-performance liquid chromatography-mass spectrometry. A tiered risk assessment approach was also considered. In the Tier 1 assessment, the mean and worst-case exposure were estimated to screen for rough exposure and risk levels. In the Tier 2 assessment, mean and upper-tail exposure levels were calculated based on the exposure parameters of Korean consumers using Monte Carlo simulation. QACs had a low frequency of detection of up to 20% in dishwashing detergents, and the contents of detected QACs varied depending on the individual samples. Based on the results of the Tier 1 assessment, BACs and DDACs posed potential health risks via inhalation and dermal routes. Tier 2 assessment suggested that the current level of oral and dermal exposure of Korean consumers to QACs in dishwashing detergents is unlikely to pose a health risk, even for upper-tail exposure groups. However, the present results suggest that spray-type DDACs may pose a health risk in the upper-tail inhalation exposure group, and further investigation is required to clarify this risk.