Empowering SARS-CoV-2 variant neutralization with a bifunctional antibody engineered with tandem heptad repeat 2 peptides.

With the global pandemic and the continuous mutations of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the need for effective and broadly neutralizing treatments has become increasingly urgent. This study introduces a novel strategy that targets two aspects simultaneously, using bifunctional antibodies to inhibit both the attachment of SARS-CoV-2 to host cell membranes and viral fusion. We developed pioneering IgG4-(HR2)4 bifunctional antibodies by creating immunoglobulin G4-based and phage display-derived human monoclonal antibodies (mAbs) that specifically bind to the SARS-CoV-2 receptor-binding domain, engineered with four heptad repeat 2 (HR2) peptides. Our in vitro experiments demonstrate the superior neutralization efficacy of these engineered antibodies against various SARS-CoV-2 variants, ranging from original SARS-CoV-2 strain to the recently emerged Omicron variants, as well as SARS-CoV, outperforming the parental mAb. Notably, intravenous monotherapy with the bifunctional antibody neutralizes a SARS-CoV-2 variant in a murine model without causing significant toxicity. In summary, this study unveils the significant potential of HR2 peptide-driven bifunctional antibodies as a potent and versatile strategy for mitigating SARS-CoV-2 infections. This approach offers a promising avenue for rapid development and management in the face of the continuously evolving SARS-CoV-2 variants, holding substantial promise for pandemic control.

Anti-pollutant effect of oleic acid against urban particulate matter is mediated via regulation of AhR- and TRPV1-mediated signaling in vitro.

Urban Particulate Matter (UPM) induces skin aging and inflammatory responses by regulating skin cells through the transient receptor potential vanilloid 1 (TRPV1). Although oleic acid, an unsaturated free fatty acid (FFA), has some functional activities, its effect on UPM-induced skin damage has not been elucidated. Here, we investigated signaling pathways on how oleic acid is involved in attenuating UPM induced cell damage. UPM treatment increased XRE-promoter luciferase activity and increased translocation of AhR to the nucleus, resulting in the upregulation of CYP1A1 gene. However, oleic acid treatment attenuated the UPM effects on AhR signaling. Furthermore, while UPM induced activation of TRPV1 and MAPKs signaling which activated the downstream molecules NFκB and AP-1, these effects were reduced by cotreatment with oleic acid. UPM-dependent generation of reactive oxygen species (ROS) and reduction of cellular proliferation were also attenuated by the treatment of oleic acid. These data reveal that cell damage induced by UPM treatment occurs through AhR signaling and TRPV1 activation which in turn activates ERK and JNK, ultimately inducing NFκB and AP-1 activation. These effects were reduced by the cotreatment of oleic acid on HaCaT cells. These suggest that oleic acid reduces UPM-induced cell damage through inhibiting both the AhR signaling and activation of TRPV1 and its downstream molecules, leading to a reduction of pro-inflammatory cytokine and recovery of cell proliferation.

Development, High-Throughput Profiling, and Biopanning of a Large Phage Display Single-Domain Antibody Library.

Immunoglobulin G-based monoclonal antibodies (mAbs) have been effective in treating various diseases, but their large molecular size can limit their penetration of tissue and efficacy in multifactorial diseases, necessitating the exploration of alternative forms. In this study, we constructed a phage display library comprising single-domain antibodies (sdAbs; or "VHHs"), known for their small size and remarkable stability, using a total of 1.6 × 109 lymphocytes collected from 20 different alpacas, resulting in approximately 7.16 × 1010 colonies. To assess the quality of the constructed library, next-generation sequencing-based high-throughput profiling was performed, analyzing approximately 5.65 × 106 full-length VHH sequences, revealing 92% uniqueness and confirming the library's diverse composition. Systematic characterization of the library revealed multiple sdAbs with high affinity for three therapeutically relevant antigens. In conclusion, our alpaca sdAb phage display library provides a versatile resource for diagnostics and therapeutics. Furthermore, the library's vast natural VHH antibody repertoire offers insights for generating humanized synthetic sdAb libraries, further advancing sdAb-based therapeutics.

Thrombopoietin receptor agonist antibody for treating chemotherapy-induced thrombocytopenia.

BACKGROUND: Thrombocytopenia is a common complication in cancer patients undergoing chemotherapy. Chemotherapy-induced thrombocytopenia (CIT) leads to dose reduction and treatment delays, lowering chemotherapy efficacy and survival rate. Thus, rapid recovery and continuous maintenance of platelet count during chemotherapy cycles are crucial in patients with CIT. Thrombopoietin (TPO) and its receptor, myeloid proliferative leukemia (MPL) protein, play a major role in platelet production. Although several MPL agonists have been developed to regulate thrombopoiesis, none have been approved for the management of CIT due to concerns regarding efficacy or safety. Therefore, the development of effective MPL agonists for treating CIT needs to be further expanded. METHODS: Anti-MPL antibodies were selected from the human combinatorial antibody phage libraries using phage display. We identified 2R13 as the most active clone among the binding antibodies via cell proliferation assay using BaF3/MPL cells. The effect of 2R13 on megakaryocyte differentiation was evaluated in peripheral blood CD34+ cells by analyzing megakaryocyte-specific differentiation markers (CD41a+ and CD42b+) and DNA ploidy using flow cytometry. The 2R13-induced platelet production was examined in 8- to 10-week-old wild-type BALB/c female mice and a thrombocytopenia mouse model established by intraperitoneal injection of 5-fluorouracil (150 mg/kg). The platelet counts were monitored twice a week over 14 days post-initiation of treatment with a single injection of 2R13, or recombinant human TPO (rhTPO) for seven consecutive days. RESULTS: We found that 2R13 specifically interacted with MPL and activated its signaling pathways. 2R13 stimulated megakaryocyte differentiation, evidenced by increasing the proportion of high-ploidy (≥ 8N) megakaryocytes in peripheral blood-CD34+ cells. The platelet count was increased by a single injection of 2R13 for up to 14 days. Injection of 5-fluorouracil considerably reduced the platelet count by day 4, which was recovered by 2R13. The platelets produced by 2R13 sustained a higher count than that achieved using seven consecutive injections of rhTPO. CONCLUSIONS: Our findings suggest that 2R13 is a promising therapeutic agent for CIT treatment.

Generation and Next-Generation Sequencing-Based Characterization of a Large Human Combinatorial Antibody Library.

Antibody phage display is a key technology for the discovery and development of target-specific monoclonal antibodies (mAbs) for use in research, diagnostics, and therapy. The construction of a high-quality antibody library, with larger and more diverse antibody repertoires, is essential for the successful development of phage display-derived mAbs. In this study, a large human combinatorial single-chain variable fragment library (1.5 × 1011 colonies) was constructed from Epstein-Barr virus-infected human peripheral blood mononuclear cells stimulated with a combination of two of the activators of human B cells, the Toll-like receptor 7/8 agonist R848 and interleukin-2. Next-generation sequencing analysis with approximately 1.9 × 106 and 2.7 × 106 full-length sequences of heavy chain variable (VH) and κ light chain variable (Vκ) domains, respectively, revealed that the library consists of unique VH (approximately 94%) and Vκ (approximately 91%) sequences with greater diversity than germline sequences. Lastly, multiple unique mAbs with high affinity and broad cross-species reactivity could be isolated from the library against two therapeutically relevant target antigens, validating the library quality. These findings suggest that the novel antibody library we have developed may be useful for the rapid development of target-specific phage display-derived recombinant human mAbs for use in therapeutic and diagnostic applications.

A Fully-Human Antibody Specifically Targeting a Membrane-Bound Fragment of CADM1 Potentiates the T Cell-Mediated Death of Human Small-Cell Lung Cancer Cells.

Small-cell lung cancer (SCLC) is the most aggressive form of lung cancer and the leading cause of global cancer-related mortality. Despite the earlier identification of membrane-proximal cleavage of cell adhesion molecule 1 (CADM1) in cancers, the role of the membrane-bound fragment of CAMD1 (MF-CADM1) is yet to be clearly identified. In this study, we first isolated MF-CADM1-specific fully human single-chain variable fragments (scFvs) from the human synthetic scFv antibody library using the phage display technology. Following the selected scFv conversion to human immunoglobulin G1 (IgG1) scFv-Fc antibodies (K103.1-4), multiple characterization studies, including antibody cross-species reactivity, purity, production yield, and binding affinity, were verified. Finally, via intensive in vitro efficacy and toxicity evaluation studies, we identified K103.3 as a lead antibody that potently promotes the death of human SCLC cell lines, including NCI-H69, NCI-H146, and NCI-H187, by activated Jurkat T cells without severe endothelial toxicity. Taken together, these findings suggest that antibody-based targeting of MF-CADM1 may be an effective strategy to potentiate T cell-mediated SCLC death, and MF-CADM1 may be a novel potential therapeutic target in SCLC for antibody therapy.

Developing a Time-Adaptive Prediction Model for Out-of-Hospital Cardiac Arrest: Nationwide Cohort Study in Korea.

BACKGROUND: Out-of-hospital cardiac arrest (OHCA) is a serious public health issue, and predicting the prognosis of OHCA patients can assist clinicians in making decisions about the treatment of patients, use of hospital resources, or termination of resuscitation. OBJECTIVE: This study aimed to develop a time-adaptive conditional prediction model (TACOM) to predict clinical outcomes every minute. METHODS: We performed a retrospective observational study using data from the Korea OHCA Registry in South Korea. In this study, we excluded patients with trauma, those who experienced return of spontaneous circulation before arriving in the emergency department (ED), and those who did not receive cardiopulmonary resuscitation (CPR) in the ED. We selected patients who received CPR in the ED. To develop the time-adaptive prediction model, we organized the training data set as ongoing CPR patients by the minute. A total of 49,669 patients were divided into 39,602 subjects for training and 10,067 subjects for validation. We compared random forest, LightGBM, and artificial neural networks as the prediction model methods. Model performance was quantified using the prediction probability of the model, area under the receiver operating characteristic curve (AUROC), and area under the precision recall curve. RESULTS: Among the three algorithms, LightGBM showed the best performance. From 0 to 30 min, the AUROC of the TACOM for predicting good neurological outcomes ranged from 0.910 (95% CI 0.910-0.911) to 0.869 (95% CI 0.865-0.871), whereas that for survival to hospital discharge ranged from 0.800 (95% CI 0.797-0.800) to 0.734 (95% CI 0.736-0.740). The prediction probability of the TACOM showed similar flow with cohort data based on a comparison with the conventional model's prediction probability. CONCLUSIONS: The TACOM predicted the clinical outcome of OHCA patients per minute. This model for predicting patient outcomes by the minute can assist clinicians in making rational decisions for OHCA patients.

Second-generation antipsychotics activate platelets in antipsychotic-naive and antipsychotic-free patients with schizophrenia: A retrospective study.

Objective: Among the risk factors for cerebrovascular/cardiovascular disease or thromboembolic events caused by the administration of second-generation antipsychotics, clinicians have mainly focused on metabolic side effects, with little interest in the effects on platelet activity. Because excessive platelet activity can increase the risk for cerebrovascular/cardiovascular disease, the aim of this study was to investigate the effect of second-generation antipsychotics on platelet activity in patients with schizophrenia. Methods: The medical records of patients with schizophrenia who were treated with second-generation antipsychotics were retrospectively reviewed. The degree of platelet activation was assessed by measuring the mean platelet component. Results: Wilcoxon signed-rank test revealed that mean platelet component levels were significantly decreased by the administration of second-generation antipsychotics (V = 20; p < 0.05), suggesting that the administration of second-generation antipsychotics may increase platelet activation. Conclusion: Because platelet activation is an additional risk factor for the occurrence of cerebrovascular/cardiovascular disease, results of this study suggest that clinicians should carefully monitor the degree of platelet activation after the administration of second-generation antipsychotics.

Negative Cellular Effects of Urban Particulate Matter on Human Keratinocytes Are Mediated by P38 MAPK and NF-κB-dependent Expression of TRPV 1.

Urban particulate matter (UPM) exerts negative effects on various human organs. Transient receptor potential vanilloid 1 (TRPV1) is a polymodal sensory transducer that can be activated by multiple noxious stimuli. This study aimed to explore the effects of the UPM 1648a on the expression of TRPV1, and its regulatory mechanisms in HaCaT cells. UPM enhanced TRPV 1 promoter-luciferase reporter activity. UPM also increased expression of the TRPV 1 gene as evidenced by increased mRNA and protein levels of TRPV 1. In addition, elucidation of the underlying mechanism behind the UPM-mediated effects on TRPV 1 expression revealed that UPM can upregulate expression of the TRPV1 gene by activating activator protein-1 (AP-1) and nuclear factor kappa B (NF-κB). The UPM treatment also altered Ca2+ influx and cell proliferation, as well as production of interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and interleukin-1ß (IL-1ß). In addition, these UPM-induced effects were attenuated by SB203580 and ammonium pyrrolidinedithiocarbamate (PDTC). However, SP600125 and PD98059 did not alter the UPM-induced effects. Taken together, these findings indicate that UPM upregulates expression of the TRPV 1 gene, which is mediated by the p38 mitogen-activated protein kinase (MAPK) and NF-κB signaling pathways and suggest that UPM is a potential irritant that can induce skin processes such as aging and inflammatory responses.

Open reduction and internal fixation for intraarticular fracture of metacarpal head.

Intraarticular fracture of the metacarpophalangeal (MP) joint presents complex problems related to the sophisticated functional aspects of the hand. Injury to the metacarpal head may have a severe effect on hand function but few studies have investigated the management of this condition. In this study, we applied open reduction and internal fixation for the displaced fracture of the metacarpal head and report the clinical and radiographic outcomes of our experience. Thirteen patients (12 men, 1 woman; mean age 21 years) were included in this study, and medical records and radiographs were reviewed retrospectively. The average follow-up period was 12.5 months. Range of motion (ROM) and Disabilities of the Arm, Shoulder, and Hand (DASH) scores were analyzed, and functional results and serial radiographs were investigated for the maintenance of articular congruity and fracture union. The injured fingers were 5 long, 4 small, 2 ring, and 2 index. Five cases were fixed with K­wires, 5 cases with headless screws, and 3 cases with screw and K­wire. The average range of injured MP joint motion was 89°, total active range of motion (TAM) was 265°, and the average DASH score was 3.8 at the last follow-up. All patients showed fracture union on the radiographs and no patient showed significant articular surface incongruence or degenerative change. Open reduction and internal fixation of the metacarpal head fracture had favorable outcomes in our study. The authors suggest accurate reduction and stable fixation for better functional results in metacarpal head fractures.

The result of surgical treatment of medial epicondylitis: analysis with more than a 5-year follow-up.

BACKGROUND: Angiofibroblastic changes of a musculotendinous origin at the medial epicondyle characterize medial epicondylitis of the elbow. Although nonsurgical treatment is the primary approach for medial epicondylitis, surgical treatment should be considered when conservative therapy fails. This study reports the results of surgical treatment of medial epicondylitis monitored for more than 5 years. METHODS: This study included 55 patients with 63 cases of medial epicondylitis between 2000 and 2010. The conservative treatment periods lasted for a minimum of 1 year, and steroid injections were administered more than twice before surgery. One surgeon conducted the surgical procedures. The Nirschl and Pettrone grades, visual analog scale (VAS) scores, Disabilities of the Arm, Shoulder and Hand (DASH) scores, Mayo Elbow Performance scores, and grip strengths were analyzed. Statistical analyses were performed using paired t tests. RESULTS: The mean VAS score improved from 8.5 to 2.4 (P <.001). Nirschl and Pettrone grades rated 43% (27 elbows) as excellent and 51% (32 elbows) as good. The Mayo Elbow Performance scores improved from 72 to 88 (P <.001) and DASH scores from 57 to 23 (P <.001). The mean grip strength of the affected side improved from 30 to 43 lb (P <.001). The mean time required to return to work and exercise was 2.8 months and 4.8 months, respectively. One case of heterotrophic ossification, which had no functional instability afterward, was seen. CONCLUSION: The results indicate that surgical treatment of medial epicondylitis could be an effective and safe treatment when conservative treatment fails.

Effects of Ganodermanondiol, a New Melanogenesis Inhibitor from the Medicinal Mushroom Ganoderma lucidum.

Ganoderma lucidum, a species of the Basidiomycetes class, has been attracting international attention owing to its wide variety of biological activities and great potential as an ingredient in skin care cosmetics including "skin-whitening" products. However, there is little information available on its inhibitory effect against tyrosinase activity. Therefore, the objectives of this study were to investigate the chemical composition of G. lucidum and its inhibitory effects on melanogenesis. We isolated the active compound from G. lucidum using ethanol extraction and ethyl acetate fractionation. In addition, we assayed its inhibitory effects on tyrosinase activity and melanin biosynthesis in B16F10 melanoma cells. In this study, we identified a bioactive compound, ganodermanondiol, which inhibits the activity and expression of cellular tyrosinase and the expression of tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor (MITF), thereby decreasing melanin production. Furthermore, ganodermanondiol also affected the mitogen-activated protein kinase (MAPK) cascade and cyclic adenosine monophosphate (cAMP)-dependent signaling pathway, which are involved in the melanogenesis of B16F10 melanoma cells. The finding that ganodermanondiol from G. lucidum exerts an inhibitory effect on tyrosinase will contribute to the use of this mushroom in the preparation of skin care products in the future.

Proximal Segment Changes After Bilateral Sagittal Split Ramus Osteotomy in Facial Asymmetry Patients.

PURPOSE: To investigate the 3-dimensional postoperative changes in the proximal segments in patients with facial asymmetry according to the anteroposterior skeletal patterns. MATERIALS AND METHODS: A total of 51 patients with facial asymmetry who had undergone Le Fort I osteotomy and sagittal split ramus osteotomy (SSRO) with rigid fixation were classified according to their anteroposterior skeletal patterns. Cone-beam computed tomography (CBCT) data were obtained before (T0) and 6 months (T1) after surgery. The position, angulation, and distance of the proximal segment were measured from the CBCT superimposition. RESULTS: Comparing the T0 and T1 CBCT data, almost no significant differences were found in the condylar head position in any of the groups (P > .05), except for the axial condylar head position on the deviated side in the skeletal Class I group (P < .05) and the sagittal condylar head position on the deviated side in the skeletal Class III group (P < .05). However, the changes in the coronoid process and ramus down varied (P < .05), and these movements were related to changes in the ramal plane. Comparing the deviated and nondeviated sides, significant differences were found only in the skeletal Class I group (P < .05). CONCLUSIONS: The most influential factors affecting differences between the deviated and nondeviated sides in patients with facial asymmetry after bilateral SSRO could be the direction of the surgical movement of the distal segment of the mandible rather than its extent.

The effect of word imagery on priming effect under a preconscious condition: an fMRI study.

Semantic priming is affected by the degree of association and how readily a word is imagined. In the association effect, activity in the perisylvian structures including the bilateral inferior frontal gyrus, the left middle temporal gyrus, and the supramarginal gyrus was correlated. However, little is known about the brain regions related to the effect of imagery word under the preconscious condition. Forty word pairs for high (HA)-, low (LA)-, and nonassociation (NA), nonword (NW) conditions were presented. Each 40 association word pairs (HA and LA) included 20 high (HI) and 20 low (LI) imagery prime stimuli, using a visually presented lexical decision task. A trial consisted of 30 ms prime, 30 ms mask, 500 ms probe, and 2-8 s stimulus onset asynchrony. Brain activation was measured using functional magnetic resonance imaging during word discrimination. Behavioral data indicated that the shortest response time (RT) was given for HA words, followed by LA and NA, and NW showed the longest RT (P < 0.01). RT was faster in HI than LI within HA, but not LA conditions (P < 0.01). Functional neuroimaging showed that differential brain regions for high imagery (HI) and low imagery (LI) words within low prime-target word association were observed in the left precuneus, left posterior cingulate gyrus, and right cuneal cortex. The present findings demonstrate that the effect of the degree of imagery on semantic priming occurs during the early stage of language processing, indicating an "automatic imagery priming effect." Our paradigm may be useful to explore semantic deficit related to imagery in various psychiatric disorders.

Autonomous Needle Navigation in Subretinal Injections via iOCT.

Subretinal injection is an effective method for direct delivery of therapeutic agents to treat prevalent subretinal diseases. Among the challenges for surgeons are physiological hand tremor, difficulty resolving single-micron scale depth perception, and lack of tactile feedback. The recent introduction of intraoperative Optical Coherence Tomography (iOCT) enables precise depth information during subretinal surgery. However, even when relying on iOCT, achieving the required micron-scale precision remains a significant surgical challenge. This work presents a robot-assisted workflow for high-precision autonomous needle navigation for subretinal injection. The workflow includes online registration between robot and iOCT coordinates; tool-tip localization in iOCT coordinates using a Convolutional Neural Network (CNN); and tool-tip planning and tracking system using real-time Model Predictive Control (MPC). The proposed workflow is validated using a silicone eye phantom and ex vivo porcine eyes. The experimental results demonstrate that the mean error to reach the user-defined target and the mean procedure duration are within an acceptable precision range. The proposed workflow achieves a 100% success rate for subretinal injection, while maintaining scleral forces at the scleral insertion point below 15mN throughout the navigation procedures.

The Effect of Intermittent Pneumatic Compression on Hemodynamics and Regional Cerebral Oxygen Saturation in Laparoscopic Bariatric Surgery with Mild Hypercapnia in the Steep Reverse Trendelenburg Position.

Obesity negatively affects hemodynamics and cerebral physiology. We investigated the effect of the utilization of an intermittent pneumatic compression (IPC) device on hemodynamics and cerebral physiology in patients undergoing laparoscopic bariatric surgery under general anesthesia with lung-protective ventilation. Sixty-four patients (body mass index > 30 kg/m2) were randomly assigned to groups that received an IPC device (IPC group, n = 32) and did not (control group, n = 32). The mean arterial pressure (MAP), heart rate (HR), need for vasopressors, cerebral oxygen saturation (rSO2), and cerebral desaturation events were recorded. The incidence of intraoperative hypotension was not significantly different between groups (p = 0.153). Changes in MAP and HR over time were similar between groups (p = 0.196 and p = 0.705, respectively). The incidence of intraoperative cerebral desaturation was not significantly different between groups (p = 0.488). Changes in rSO2 over time were similar between the two groups (p = 0.190) during pneumoperitoneum. Applying IPC to patients with obesity in the steep reverse Trendelenburg position may not improve hemodynamic parameters, vasopressor requirements, or rSO2 values during pneumoperitoneum under lung-protective ventilation. During laparoscopic bariatric surgery, IPC alone has limitations in improving hemodynamics and cerebral physiology.

Targeting cell surface glucose-regulated protein 94 in gastric cancer with an anti-GRP94 human monoclonal antibody.

Gastric cancer (GC), a leading cause of cancer-related mortality, remains a significant challenge despite recent therapeutic advancements. In this study, we explore the potential of targeting cell surface glucose-regulated protein 94 (GRP94) with antibodies as a novel therapeutic approach for GC. Our comprehensive analysis of GRP94 expression across various cancer types, with a specific focus on GC, revealed a substantial overexpression of GRP94, highlighting its potential as a promising target. Through in vitro and in vivo efficacy assessments, as well as toxicological analyses, we found that K101.1, a fully human monoclonal antibody designed to specifically target cell surface GRP94, effectively inhibits GC growth and angiogenesis without causing in vivo toxicity. Furthermore, our findings indicate that K101.1 promotes the internalization and concurrent downregulation of cell surface GRP94 on GC cells. In conclusion, our study suggests that cell surface GRP94 may be a potential therapeutic target in GC, and that antibody-based targeting of cell surface GRP94 may be an effective strategy for inhibiting GRP94-mediated GC growth and angiogenesis. [BMB Reports 2024; 57(4): 188-193].

A dual-targeting approach using a human bispecific antibody against the receptor-binding domain of the Middle East Respiratory Syndrome Coronavirus.

The emergence of the Middle East Respiratory Syndrome Coronavirus (MERS-CoV) has posed a significant global health concern due to its severe respiratory illness and high fatality rate. Currently, despite the potential for resurgence, there are no specific treatments for MERS-CoV, and only supportive care is available. Our study aimed to address this therapeutic gap by developing a potent neutralizing bispecific antibody (bsAb) against MERS-CoV. Initially, we isolated four human monoclonal antibodies (mAbs) that specifically target the MERS-CoV receptor-binding domain (RBD) using phage display technology and an established human antibody library. Among these four selected mAbs, our intensive in vitro functional analyses showed that the MERS-CoV RBD-specific mAb K111.3 exhibited the most potent neutralizing activity against MERS-CoV pseudoviral infection and the molecular interaction between MERS-CoV RBD and human dipeptidyl peptidase 4. Consequently, we engineered a novel bsAb, K207.C, by utilizing K111.3 as the IgG base and fusing it with the single-chain variable fragment of its non-competing pair, K111.1. This engineered bsAb showed significantly enhanced neutralization potential against MERS-CoV compared to its parental mAb. These findings suggest that K207.C may serve as a potential candidate for effective MERS-CoV neutralization, further highlighting the promise of the bsAb dual-targeting approach in MERS-CoV neutralization.

Autonomous Needle Navigation in Retinal Microsurgery: Evaluation in ex vivo Porcine Eyes.

Important challenges in retinal microsurgery include prolonged operating time, inadequate force feedback, and poor depth perception due to a constrained top-down view of the surgery. The introduction of robot-assisted technology could potentially deal with such challenges and improve the surgeon's performance. Motivated by such challenges, this work develops a strategy for autonomous needle navigation in retinal microsurgery aiming to achieve precise manipulation, reduced end-to-end surgery time, and enhanced safety. This is accomplished through real-time geometry estimation and chance-constrained Model Predictive Control (MPC) resulting in high positional accuracy while keeping scleral forces within a safe level. The robotic system is validated using both open-sky and intact (with lens and partial vitreous removal) ex vivo porcine eyes. The experimental results demonstrate that the generation of safe control trajectories is robust to small motions associated with head drift. The mean navigation time and scleral force for MPC navigation experiments are 7.208 s and 11.97 mN, which can be considered efficient and well within acceptable safe limits. The resulting mean errors along lateral directions of the retina are below 0.06 mm, which is below the typical hand tremor amplitude in retinal microsurgery.

Development and Characterization of Phage Display-Derived Monoclonal Antibodies to the S2 Domain of Spike Proteins of Wild-Type SARS-CoV-2 and Multiple Variants.

The rapid emergence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants has resulted in the ongoing global coronavirus disease 2019 (COVID-19) pandemic. Thus, the rapid development of a platform to detect a broad range of SARS-CoV-2 variants is essential for successful COVID-19 management. In this study, four SARS-CoV-2 spike protein-specific single-chain variable fragments (scFvs) were isolated from a synthetic antibody library using phage display technology. Following the conversion of these scFvs into monoclonal antibodies (mAbs) (K104.1-K104.4) and production and purification of the mAbs, the antibody pair (K104.1 and K104.2) that exhibited the highest binding affinity (K104.1 and K104.2, 1.3 nM and 1.9 nM) was selected. Biochemical analyses revealed that this antibody pair specifically bound to different sites on the S2 subunit of the spike protein. Furthermore, we developed a highly sensitive sandwich immunoassay using this antibody pair that accurately and quantitatively detected the spike proteins of wild-type SARS-CoV-2 and multiple variants, including Alpha, Beta, Gamma, Delta, Kappa, and Omicron, in the picomolar range. Conclusively, the novel phage display-derived mAbs we have developed may be useful for the rapid and efficient detection of the fast-evolving SARS-CoV-2.