Pharmacokinetics and safety of LEAD-452, an EGFR-specific 4-1BB-agonistic trimerbody in non-human primates.

LEAD-452 is a humanized bispecific EGFR-targeted 4-1BB-agonistic trimerbody with a unique trimeric configuration compared to other 4-1BB-specific antibodies that are currently in development. Indeed, enhanced tumor-specific costimulation and very remarkable safety and efficacy profiles have been observed in mouse models. Here, we conducted for the first time a preclinical pharmacokinetic and toxicity study in non-human primates (NHP) (Macaca fascicularis). LEAD-452 exhibits comparable binding affinity for human and macaque targets, indicating its pharmacological significance for safety testing across species. The NHP were administered LEAD-452 in a series of ascending doses, ranging from 0.1 mg/kg to 10 mg/kg, and repeated doses up to 20 mg/kg. The administration of LEAD-452 was found to be clinically well tolerated, with no major related adverse effects observed. Furthermore, there have been no reported cases of liver toxicity, thrombocytopenia, and neutropenia, which are commonly associated with treatments using conventional anti-4-1BB IgG-based antibodies. In addition, neither IgM nor IgG-based anti-drug antibodies were detected in serum samples from NHP during the study, regardless of the dose of LEAD-452 administered. These results support the clinical development of LEAD-452 for the treatment of solid tumors.

Adaptación y desempeño psicométrico de la Escala de tormento físico y emocional hacia animales.

Introducción: El tormento físico y emocional hacia los animales durante la infancia y la adultez emergente ha sido relacionado con el diagnóstico de diversos trastornos de la conducta. El objetivo de este estudio fue adaptar y validar la Escala de tormento físico y emocional hacia animales (PET) en una muestra de universitarios del Caribe colombiano. Materiales y método: Estudio cuantitativo de carácter instrumental, con enfoque transver sal. Se evaluaron exploratoriamente tres modelos en 100 universitarios y se confirmaron dos soluciones del instrumento en 140 universitarios. Resultados: En el análisis factorial exploratorio (AFE), dos modelos cumplieron con la asignación de factores; el primero produjo una estructura factorial bidimensional que explicó el 68.39% de la varianza de los datos, con esfericidad de Bartlett significativa (331,099, gl=36, Sig.=.002), indicador de adecuación del tamaño de muestra Kaiser-Meyer-Olkin aceptable (KMO=.73) y consistencia interna total y dimensional adecuada ( ɑ ordinal total = .825; ɑ ordina subescala 1 = .93 y ordinal subescala 2 = .75). El segundo reportó una estructura factorial unidimensional que explicó el 73.16% e indicadores de esfericidad,tamaño de muestra y consistencia adecuados (216.369, gl=6, Sig.=.000; KMO=.73 y ɑ ordinal total = .877). Las soluciones exploradas fueron confirmadas encontrando para ambos modelos indicadores de bondad del ajuste satisfactorios (modelo bidimensional: X 2 ɑ /gl=2.33; GFI=.991; AGFI=.969; CFI=.944; NFI=.970; RMSEA=.070 y SRMR=.070. modelo unidimensional: X /gl=1.98; GFI=.998; AGFI=.987; CFI=.968; NFI=.994; RMSEA=.068 y SRMR=.040). Discusión y conclusiones: La escala PET en su versión original y reducida es un instrumento válido y confiable para evaluar este constructo en el contexto sociocultural colombiano.

MEIoT 2D-CACSET: IoT Two-Dimensional Cartesian Coordinate System Educational Toolkit Align with Educational Mechatronics Framework.

The educational sector has made extraordinary efforts to neutralize the impact of the pandemic caused by COVID-19, forcing teachers, scholars, and academic personnel to change the way education is delivered by developing creative and technological solutions to improve the landscape for education. The Internet of Things (IoT) is crucial for the educational transition to digital and virtual environments. This paper presents the integration of IoT technology in the Two-Dimensional Cartesian Coordinate System Educational Toolkit (2D-CACSET), to transform it into MEIoT 2D-CACSET; which includes educational mechatronics and the IoT. The Educational Mechatronics Conceptual Framework (EMCF) is extended to consider the virtual environment, enabling knowledge construction in virtual concrete, virtual graphic, and virtual abstract levels. Hence, the students acquire this knowledge from a remote location to apply it further down their career path. Three instructional designs are designed for this work using the MEIoT 2D-CACSET to learn about coordinate axes, quadrants, and a point in the 2D Coordinate Cartesian System. This work is intended to provide an IoT educational technology to offer an adequate response to the educational system's current context.

Methodology for the Development of Augmented Reality Applications: MeDARA. Drone Flight Case Study.

Industry 4.0 involves various areas of engineering such as advanced robotics, Internet of Things, simulation, and augmented reality, which are focused on the development of smart factories. The present work presents the design and application of the methodology for the development of augmented reality applications (MeDARA) using a concrete, pictorial, and abstract approach with the intention of promoting the knowledge, skills, and attitudes of the students within the conceptual framework of educational mechatronics (EMCF). The flight of a drone is presented as a case study, where the concrete level involves the manipulation of the drone in a simulation; the graphic level requires the elaboration of an experiential storyboard that shows the scenes of the student's interaction with the drone in the concrete level; and finally, the abstract level involves the planning of user stories and acceptance criteria, the computer design of the drone, the mock-ups of the application, the coding in Unity and Android Studio, and its integration to perform unit and acceptance tests. Finally, evidence of the tests is shown to demonstrate the results of the application of the MeDARA.

UAV-Based Smart Educational Mechatronics System Using a MoCap Laboratory and Hardware-in-the-Loop.

Within Industry 4.0, drones appear as intelligent devices that have brought a new range of innovative applications to the industrial sector. The required knowledge and skills to manage and appropriate these technological devices are not being developed in most universities. This paper presents an unmanned aerial vehicle (UAV)-based smart educational mechatronics system that makes use of a motion capture (MoCap) laboratory and hardware-in-the-loop (HIL) to teach UAV knowledge and skills, within the Educational Mechatronics Conceptual Framework (EMCF). The macro-process learning construction of the EMCF includes concrete, graphic, and abstract levels. The system comprises a DJI Phantom 4, a MoCap laboratory giving the drone location, a Simulink drone model, and an embedded system for performing the HIL simulation. The smart educational mechatronics system strengthens the assimilation of the UAV waypoint navigation concept and the capacity for drone flight since it permits the validation of the physical drone model and testing of the trajectory tracking control. Moreover, it opens up a new range of possibilities in terms of knowledge construction through best practices, activities, and tasks, enriching the university courses.

Extracorporeal CO2 removal and renal replacement therapy in acute severe respiratory failure in COVID-19 pneumonia: Case report.

The COVID-19 pandemic significates an enormous number of patients with pneumonia that get complicated with severe acute respiratory distress syndrome (ARDS), some of them with refractory hypercapnia and hypoxemia that need mechanical ventilation (MV). Those patients who are not candidate to extracorporeal membrane oxygenation (ECMO), the extracorporeal removal of CO2 (ECCO2 R) can allow ultra protective MV to limit the transpulmonary pressures and avoid ventilatory induced lung injury (VILI). We report a first case of prolonged ECCO2 R support in 38 year male with severe COVID-19 pneumonia refractory to conventional support. He was admitted tachypneic and oxygen saturation 71% without supplementary oxygen. The patient's clinical condition worsens with severe respiratory failure, increasing the oxygen requirement and initiating MV in the prone position. After 21 days of protective MV, PaCO2 rise to 96.8 mmHg, making it necessary to connect to an ECCO2 R system coupled continuous veno-venous hemodialysis (CVVHD). However, due to the lack of availability of equipment in the context of the pandemic, a pediatric gas exchange membrane adapted to CVVHD allowed to maintain the removal of CO2 until completing 27 days, being finally disconnected from the system without complications and with a satisfactory evolution.

Implementation of a MEIoT Weather Station with Exogenous Disturbance Input.

Due to the emergence of the coronavirus disease (COVID 19), education systems in most countries have adapted and quickly changed their teaching strategy to online teaching. This paper presents the design and implementation of a novel Internet of Things (IoT) device, called MEIoT weather station, which incorporates an exogenous disturbance input, within the National Digital Observatory of Smart Environments (OBNiSE) architecture. The exogenous disturbance input involves a wind blower based on a DC brushless motor. It can be controlled, via Node-RED platform, manually through a sliding bar, or automatically via different predefined profile functions, modifying the wind speed and the wind vane sensor variables. An application to Engineering Education is presented with a case study that includes the instructional design for the least-squares regression topic for linear, quadratic, and cubic approximations within the Educational Mechatronics Conceptual Framework (EMCF) to show the relevance of this proposal. This work's main contribution to the state-of-the-art is to turn a weather monitoring system into a hybrid hands-on learning approach thanks to the integrated exogenous disturbance input.

Design and Implementation of the E-Switch for a Smart Home.

As the development of systems in smart homes is increasing, it is of ever-increasing importance to have data, which artificial intelligence methods and techniques can apply to recognize activities and patterns or to detect anomalies, with the aim of reducing energy consumption in the main home domestic services, and to offer users an alternative in the management of these resources. This paper describes the design and implementation of a platform based on the internet of things and a cloud environment that allows the user to remotely control and monitor Wi-Fi wireless e-switch in a home through a mobile application. This platform is intended to represent the first step in transforming a home into a smart home, and it allows the collection and storage of the e-switch information, which can be used for further processing and analysis.

Role of nucleotide-binding oligomerization domain 1 (NOD1) in pericyte-mediated vascular inflammation.

We have recently described the response of human brain pericytes to lipopolysaccharide (LPS) through toll-like receptor 4 (TLR4). However, Gram-negative pathogen-associated molecular patterns include not only LPS but also peptidoglycan (PGN). Given that the presence of co-purified PGN in the LPS preparation previously used could not be ruled out, we decided to analyse the expression of the intracellular PGN receptors NOD1 and NOD2 in HBP and compare the responses to their cognate agonists and ultrapure LPS. Our findings show for the first time that NOD1 is expressed in pericytes, whereas NOD2 expression is barely detectable. The NOD1 agonist C12-iE-DAP induced IL6 and IL8 gene expression by pericytes as well as release of cytokines into culture supernatant. Moreover, we demonstrated the synergistic effects of NOD1 and TLR4 agonists on the induction of IL8. Using NOD1 silencing in HBP, we showed a requirement for C12-iE-DAP-dependent signalling. Finally, we could discriminate NOD1 and TLR4 pathways in pericytes by pharmacological targeting of RIPK2, a kinase involved in NOD1 but not in TLR4 signalling cascade. p38 MAPK and NF-κB appear to be downstream mediators in the NOD1 pathway. In summary, these results indicate that pericytes can sense Gram-negative bacterial products by both NOD1 and TLR4 receptors, acting through distinct pathways. This provides new insight about how brain pericytes participate in the inflammatory response and may have implications for disease management.

Hybrid Cardiac Rehabilitation Program in a Low-Resource Setting: A Randomized Clinical Trial.

Importance: While effective, cardiovascular rehabilitation (CR) as traditionally delivered is not well implemented in lower-resource settings. Objective: To test the noninferiority of hybrid CR compared with traditional CR in terms of cardiovascular events. Design, Setting, and Participants: This pragmatic, multicenter, parallel arm, open-label randomized clinical trial (the Hybrid Cardiac Rehabilitation Trial [HYCARET]) with blinded outcome assessment was conducted at 6 referral centers in Chile. Adults aged 18 years or older who had a cardiovascular event or procedure, no contraindications to exercise, and access to a mobile telephone were eligible and recruited between April 1, 2019, and March 15, 2020, with follow-up until July 29, 2021. Interventions: Participants were randomized 1:1 in permuted blocks to the experimental arm, which received 10 center-based supervised exercise sessions plus counseling in 4 to 6 weeks and then were supported at home via telephone calls and text messages through weeks 8 to 12, or the control arm, which received the standard CR of 18 to 22 sessions with exercises and education in 8 to 12 weeks. Main Outcomes and Measures: The primary outcome was cardiovascular events or mortality. Secondary outcomes were quality of life, return to work, and lifestyle behaviors measured with validated questionnaires; muscle strength and functional capacity, measured through physical tests; and program adherence and exercise-related adverse events, assessed using checklists. Results: A total of 191 participants were included (mean [SD] age, 58.74 [9.80] years; 145 [75.92%] male); 93 were assigned to hybrid CR and 98 to standard CR. At 1 year, events had occurred in 5 unique participants in the hybrid CR group (5.38%) and 9 in the standard CR group (9.18%). In the intention-to-treat analysis, the hybrid CR group had 3.80% (95% CI, -11.13% to 3.52%) fewer cardiovascular events than the standard CR group, and relative risk was 0.59 (95% CI, 0.20-1.68) for the primary outcome. In the per-protocol analysis at different levels of adherence to the intervention, all 95% CIs crossed the noninferiority boundary (eg, 20% adherence: absolute risk difference, -0.35% [95% CI, -7.56% to 6.85%]; 80% adherence: absolute risk difference, 3.30% [95% CI, -3.70% to 10.31%]). No between-group differences were found for secondary outcomes except adherence to supervised CR sessions (79.14% [736 of 930 supervised sessions] in the hybrid CR group vs 61.46% [1201 of 1954 sessions] in the standard CR group). Conclusions and Relevance: The results suggest that a hybrid CR program is noninferior to standard center-based CR in a low-resource setting, primarily in terms of recurrent cardiovascular events and potentially in terms of intermediate outcomes. Hybrid CR may induce superior adherence to supervised exercise. Clinical factors and patient preferences should inform CR model allocation. Trial Registration: ClinicalTrials.gov Identifier: NCT03881150.

Characterization of a Trispecific PD-L1 Blocking Antibody That Exhibits EGFR-Conditional 4-1BB Agonist Activity.

Immune checkpoint blockade has changed the treatment paradigm for advanced solid tumors, but the overall response rates are still limited. The combination of checkpoint blockade with anti-4-1BB antibodies to stimulate tumor-infiltrating T cells has shown anti-tumor activity in human trials. However, the further clinical development of these antibodies has been hampered by significant off-tumor toxicities. Here, we generated an anti-4-1BB/EGFR/PD-L1 trispecific antibody consisting of a triple-targeting tandem trimerbody (TT) fused to an engineered silent Fc region. This antibody (IgTT-4E1-S) was designed to combine the blockade of the PD-L1/PD-1 axis with conditional 4-1BB costimulation specifically confined to the tumor microenvironment (TME). The antibody demonstrated simultaneous binding to purified EGFR, PD-L1, and 4-1BB in solution, effective blockade of the PD-L1/PD1 interaction, and potent 4-1BB-mediated costimulation, but only in the presence of EGFR-expressing cells. These results demonstrate the feasibility of IgTT-4E1-S specifically blocking the PD-L1/PD-1 axis and inducing EGFR-conditional 4-1BB agonist activity.

Treatment of gaseous streams polluted with H2S: Comparison of electrolytic and electro-Fenton assisted absorption processes.

H2S is a gaseous compound that contributes to air pollution. In this work, the electrochemical oxidation treatment of gaseous streams polluted with H2S is evaluated using a jet mixer and electrochemical cell device, in which the performance of electrolytic and electro-Fenton assisted absorption processes are compared. Results demonstrate the feasibility of both processes to remove H2S, reaching coulombic efficiencies of nearly 100% in the electrolytic assisted absorption, and 70-80% in the electro-Fenton assisted absorption. Aqueous solutions containing phosphate salts as electrolyte were found to be suitable as absorbents for the process. Efficiency in the cathodic production of H2O2 in these solutions using the experimental device was found to be as high as 32.8% (1.184 mgH2O2/min) at 12 °C and atmospheric pressure. Sequential formation of SO2 and SO3 is obtained by the oxidation of H2S contained in the gas. These species are hydrolysed, and a part remained in the absorbent as SO32- and SO42-, while the rest is dragged in the outlet gas. SO3 production is promoted by electrolytic assisted absorption and polysulphides by the electro-Fenton technology. Low concentrations of elemental sulphur are detected in the solid suspensions formed during the process.

A PD-L1/EGFR bispecific antibody combines immune checkpoint blockade and direct anti-cancer action for an enhanced anti-tumor response.

Immune checkpoint blockade (ICB) with antibodies has shown durable clinical responses in a wide range of cancer types, but the overall response rate is still limited. Other effective therapeutic modalities to increase the ICB response rates are urgently needed. New bispecific antibody (bsAb) formats combining the ICB effect and a direct action on cancer cells could improve the efficacy of current immunotherapies. Here, we report the development of a PD-L1/EGFR symmetric bsAb by fusing a dual-targeting tandem trimmer body with the human IgG1 hinge and Fc regions. The bsAb was characterized in vitro and the antitumor efficacy was evaluated in humanized mice bearing xenografts of aggressive triple-negative breast cancer and lung cancer. The IgG-like hexavalent bsAb, designated IgTT-1E, was able to simultaneously bind both EGFR and PD-L1 antigens, inhibit EGF-mediated proliferation, effectively block PD-1/PD-L1 interaction, and induce strong antigen-specific antibody-dependent cellular cytotoxicity activity in vitro. Potent therapeutic efficacies of IgTT-1E in two different humanized mouse models were observed, where tumor growth control was associated with a significantly increased proportion of CD8+ T cells. These results support the development of IgTT-1E for the treatment of EGFR+ cancers.

Dendritic Cell-Mediated Cross-Priming by a Bispecific Neutralizing Antibody Boosts Cytotoxic T Cell Responses and Protects Mice against SARS-CoV-2.

Administration of neutralizing antibodies (nAbs) has proved to be effective by providing immediate protection against SARS-CoV-2. However, dual strategies combining virus neutralization and immune response stimulation to enhance specific cytotoxic T cell responses, such as dendritic cell (DC) cross-priming, represent a promising field but have not yet been explored. Here, a broadly nAb, TNT , are first generated by grafting an anti-RBD biparatopic tandem nanobody onto a trimerbody scaffold. Cryo-EM data show that the TNT structure allows simultaneous binding to all six RBD epitopes, demonstrating a high-avidity neutralizing interaction. Then, by C-terminal fusion of an anti-DNGR-1 scFv to TNT , the bispecific trimerbody TNT DNGR-1 is generated to target neutralized virions to type 1 conventional DCs (cDC1s) and promote T cell cross-priming. Therapeutic administration of TNT DNGR-1, but not TNT , protects K18-hACE2 mice from a lethal SARS-CoV-2 infection, boosting virus-specific humoral responses and CD8+ T cell responses. These results further strengthen the central role of interactions with immune cells in the virus-neutralizing antibody activity and demonstrate the therapeutic potential of the Fc-free strategy that can be used advantageously to provide both immediate and long-term protection against SARS-CoV-2 and other viral infections.

Voluntary self-initiation of the stimuli onset improves working memory and accelerates visual and attentional processing.

The ability of an organism to voluntarily control the stimuli onset modulates perceptual and attentional functions. Since stimulus encoding is an essential component of working memory (WM), we conjectured that controlling the initiation of the perceptual process would positively modulate WM. To corroborate this proposition, we tested twenty-five healthy subjects in a modified-Sternberg WM task under three stimuli presentation conditions: an automatic presentation of the stimuli, a self-initiated presentation of the stimuli (through a button press), and a self-initiated presentation with random-delay stimuli onset. Concurrently, we recorded the subjects' electroencephalographic signals during WM encoding. We found that the self-initiated condition was associated with better WM accuracy, and earlier latencies of N1, P2 and P3 evoked potential components representing visual, attentional and mental review of the stimuli processes, respectively. Our work demonstrates that self-initiated stimuli enhance WM performance and accelerate early visual and attentional processes deployed during WM encoding. We also found that self-initiated stimuli correlate with an increased attentional state compared to the other two conditions, suggesting a role for temporal stimuli predictability. Our study remarks on the relevance of self-control of the stimuli onset in sensory, attentional and memory updating processing for WM.

A hybrid exercise-based cardiac rehabilitation program is an effective strategy to improve muscle strength and functional exercise capacity in adults and older people with coronary artery disease.

Coronary heart disease is the most common cause of death worldwide. Standard cardiac rehabilitation (face-to-face sessions) has shown benefits in increasing muscle strength and functional exercise capacity in adults and older people. However, it is unknown whether hybrid cardiac rehabilitation (a first face-to-face phase + a second remote monitoring phase) will have similar benefits in adults versus older subjects. The aim of this study was to compare the effects of a hybrid exercise-based cardiac rehabilitation program on muscle strength and functional exercise capacity in "adult" versus "older" people with coronary artery disease. We hypothesized that a hybrid exercise-based cardiac rehabilitation program would improve muscle strength and functional exercise capacity, but the impact would be smaller in the older group than the adult individuals. This study is part of a larger project (The Hybrid Cardiac Rehabilitation Trial-HYCARET). We subjected 22 adult (<60 y) females and males (ADULT; n = 5/17 (f/m); 52 ± 5 y; 28.9 ± 3.4 kg·m-2) and 20 older (≥60 y) females and males (OLDER; n = 6/14 (f/m); 66 ± 4 y; 27.4 ± 3.9 kg·m-2) with coronary artery disease to 12 weeks of hybrid exercise-based cardiac rehabilitation program. Prior to and after 12 weeks of a hybrid exercise-based cardiac rehabilitation program, grip strength (handgrip), leg strength (chair stand test), and functional exercise capacity (6-minute walk test, 6MWT) were assessed. The hybrid exercise-based cardiac rehabilitation program resulted in a 9.4 ± 14.6% and a 6.2 ± 12.1% grip strength increase, a 14.4 ± 39.4% and a 28.9 ± 48.1% legs strength increase, and a 14.6 ± 26.4% and a 6.8 ± 14.0% functional exercise capacity improvement in ADULT and OLDER, respectively (p < 0.05) with no differences between groups. In conclusion, a hybrid exercise-based cardiac rehabilitation program could increase muscle strength and improve functional exercise capacity in adults and older people with coronary artery disease. More future studies comparing effectiveness among these age groups are needed to strengthen this conclusion.

Two-Minute Step Test as a Complement to Six-Minute Walk Test in Subjects With Treated Coronary Artery Disease.

The 2-Minute Step Test (2MST) has been presented as an alternative to the 6-Minute Walk Test (6MWT) based on the association between the two tests in older adults; however, some authors propose that it should not be a substitute but rather a complement to the latter in the fitness evaluation. Specifically, in coronary disease, despite the potential and clinical utility of 2MST, the relationship of both tests in this population is unknown. This study aimed to determine the relationship between 6MWT and 2MST and to explore the relationship of biodemographic factors for both tests in subjects with treated coronary artery disease. For this, the 6MWT and the 2MST were applied to patients with coronary artery disease treated in 6 hospitals in Chile between May 2019 and February 2020. Additionally, lower limb strength was assessed by a chair-stand test, grip strength was assessed by a dynamometer, and physical measurements were applied. In total, 163 participants underwent both tests (average age = 58.7 ± 9.8 years; 73.6% men; 64.4% revascularized by angioplasty; 28.2% revascularized by surgery, and 7.4% treated by drugs or thrombolysis). Heart rate was higher at the end of the 6MWT, while the perception of effort was greater at the end of the 2MST. There was a weak positive correlation between the 6MWT and the 2MST in subjects with treated coronary disease (r = 0.28, p = 0.0003). While age (r = -0.27), weight (r = 0.25), height (r = 0.49), and strength of both lower limbs (r = 0.41) and grip strength (r = 0.53) correlated weakly or moderately to the covered distance in 6MWT, the number of steps by the 2MST correlated only weakly to height (r = 0.23), lower limb strength (r = 0.34), and grip strength (r = 0.34). Age, weight, height, lower limb strength, and grip strength would explain better the meters walked in the 6MWT than the steps achieved in the 2MST. With these findings, we can conclude that, in patients with treated coronary artery disease, it does not seem advisable to replace 6MWT with 2MST when it is possible to do so. Additionally, the 2MST may provide additional information in the fitness evaluation. However, the usefulness of 2MST in this population needs to be further studied.

Digital physiotherapy intervention in children in a low resource setting in Anantapur (India): Study protocol for a randomized controlled trial.

Introduction: In rural India the scarcity of physiotherapists and inequalities complicate the recovery of traumatized children. This study protocol will explore a digital physiotherapy intervention in children with ankle fracture in a low-resource setting to improve functional independence and quality of life. Methods and analysis: A randomized clinical trial with a mixed quantitative-qualitative design will be carried out. It is a single-blind study, where the evaluator does not know the nature of the intervention. Sixty subjects will be enrolled and randomly divided into two groups: the experimental group (EG) will receive a 4-week digital physiotherapy intervention through an app in a recycled mobile device after hospital discharge; the control group (CG) will receive the physiotherapy standard care recommended for patients discharged from the hospital. Subjects will receive a baseline (T0-pre) assessment of Functional Independence and Quality of Life. At the end of the 4-week intervention (T1-post) a new assessment of the outcome will be performed adding data on adherence, satisfaction (ad hoc questionnaire and TSQ), and barriers of use. Qualitative outcomes will also be explored. The author's hypothesized that the implementation of a digital physiotherapy intervention is feasible and effective to improve functional independence and quality of life. This study protocol is the first to explore the effect of digital physiotherapy intervention in children's patients in a low resource setting (Anantapur). Discussion: The successful delivery of the intervention, an optimal adherence records, the absence of significant adverse effects, user satisfaction level and the qualitative analysis of limitations, will demonstrate the effectiveness of these procedure. This study will add more evidence in support the use of digital physiotherapy practice as an effective tool. User particularities, provider's capacity, technological and cultural limitations, and considerations for vulnerable populations will be taken into account. Clinical trial registration: NCT04946695 (https://clinicaltrials.gov/).

Trispecific T-cell engagers for dual tumor-targeting of colorectal cancer.

Retargeting of T lymphocytes toward cancer cells by bispecific antibodies has demonstrated its therapeutic potential, with one such antibody approved for the treatment of acute lymphoblastic leukemia (blinatumomab) and several other in clinical trials. However, improvement of their efficacy and selectivity for solid tumors is still required. Here, we describe a novel tandem T-cell recruiting trispecific antibody for the treatment of colorectal cancer (CRC). This construct, termed trispecific T-cell engager (TriTE), consists of a CD3-specific single-chain Fv (scFv) flanked by anti-epidermal growth factor receptor (EGFR) and anti-epithelial cell adhesion molecule (EpCAM) single-domain VHH antibodies. The TriTE was well expressed in mammalian and yeast cells, bound the cognate antigens of the three parental antibodies, and enabled the specific cytolysis of EGFR- and/or EpCAM-expressing cancer cells, without inducing T cell activation and cytoxicity against double-negative (EGFR-EpCAM-) cancer cells. Bivalent bispecific targeting of double-positive HCT116 cells by TriTE improved in vitro potency up to 100-fold compared to single-positive cells and significantly prolonged survival in vivo. In addition, it was less efficient at killing single-positive target cells than the corresponding bispecific controls, leading to potentially enhanced tumor specificity. Moreover, dual targeting of two tumor-associated antigens may contribute toward preventing the tumor escape by antigen loss caused by selective pressures from conventional single-targeting T-cell engagers, and may help to overcome antigenic heterogeneity.

Tumor targeted 4-1BB agonist antibody-albumin fusions with high affinity to FcRn induce anti-tumor immunity without toxicity.

Costimulation of tumor-infiltrating T lymphocytes by anti-4-1BB monoclonal antibodies (mAbs) has shown anti-tumor activity in human trials, but can be associated with significant off-tumor toxicities involving FcγR interactions. Here, we introduce albumin-fused mouse and human bispecific antibodies with clinically favorable pharmacokinetics designed to confine 4-1BB costimulation to the tumor microenvironment. These Fc-free 4-1BB agonists consist of an EGFR-specific VHH antibody, a 4-1BB-specific scFv, and a human albumin sequence engineered for high FcRn binding connected in tandem (LiTCo-Albu). We demonstrate in vitro cognate target engagement, EGFR-specific costimulatory activity, and FcRn-driven cellular recycling similar to non-fused FcRn high-binding albumin. The mouse LiTCo-Albu exhibited a prolonged circulatory half-life and in vivo tumor inhibition, with no indication of 4-1BB mAb-associated toxicity. Furthermore, we show a greater therapeutic effect when used in combination with PD-1-blocking mAbs. These findings demonstrate the feasibility of tumor-specific LiTCo-Albu antibodies for safe and effective costimulatory strategies in cancer immunotherapy.