James W. Larrick

Background: Excessive accumulation of fibrillar collagen causes pathological scarring and fibrosis. A promising anti-fibrotic strategy targets the extracellular assembly of collagen fibrils rather than intracellular synthesis pathways. We previously developed a chimeric monoclonal anti-collagen antibody (ACA) that targets the C-terminal telopeptide of the α2(I) chain (α2Ct) of human collagen I and effectively disrupts fibrillogenesis. This study details the engineering of a humanized antibody variant optimized for therapeutic application, augmented with a collagen-binding peptide (CBP) to enhance targeted retention in fibrotic tissues. Methods: A humanized ACA was engineered using in silico homology modeling, complementarity-determining region grafting, and sequence optimization to eliminate chemical liabilities. Variants were expressed in mammalian cells and evaluated for binding kinetics and specificity. To improve spatial localization, the CBP was fused to the antibody. The lead variant was assessed for in vitro cytotoxicity and matrix retention. In addition, small-group pilot studies were performed in a rabbit model of post-traumatic knee arthrofibrosis to provide basic information on the efficacy of the selected antibody construct. Results: The humanized ACA variants maintained high specificity and affinity for the α2Ct target. Fusing the CBP to the C-terminus of the light chain generated a construct (C-cbpACA) that enhanced matrix retention in cell culture conditions without compromising target engagement or causing cytotoxicity. In the rabbit arthrofibrosis model, intra-articular C-cbpACA delivery was associated with reduced flexion contracture and decreased total collagen deposition in the joint capsule relative to historical cohorts. Conclusions: These findings characterize a humanized, matrix-targeted ACA variant and provide preliminary support for its further evaluation in post-traumatic arthrofibrosis, though this small pilot study does not establish definitive superiority in efficacy or safety.

Background: Periorbital measurements such as margin to reflex distances, palpebral fissure height, and scleral show are critical in diagnosing and managing conditions like ptosis and disorders of the eyelid. However, deployment of automated periorbital measurement algorithms in structured research workflows remains limited by the lack of integrated capture and data management infrastructure. Objective: We developed and evaluated Glorbit, a lightweight, browser-based application for automated periorbital distance measurement using artificial intelligence (AI). The objective was to evaluate end-to-end workflow feasibility of the platform under simulated, operator-run conditions. Methods: The application integrates a DeepLabV3 segmentation model into a modular image processing pipeline with secure, site-specific Google Cloud storage, supporting local preprocessing and cloud upload through Firebase-authenticated logins. The full workflow—metadata entry, facial image capture, segmentation, and upload—was tested. After the session, the participants completed a Likert-style survey. Results: Glorbit successfully ran on all tested platforms, including laptops, tablets, and mobile phones across major browsers. A total of 15 volunteers were enrolled in this study in which the app completed predefined workflow steps in all simulated, operator-run sessions. The segmentation model produced outputs on all images, and the average session duration was 101.7 (SD 17.5) seconds. Simulated experience scores on a 5-point Likert scale were uniformly high. Conclusions: Glorbit is a cross-platform application that supports structured periorbital image capture and automated inference within a unified workflow. In simulated, operator-run testing, the platform demonstrated successful execution of predefined workflow steps across devices. These findings support the technical feasibility of the system as a research-oriented data collection framework and may inform future evaluations in broader research settings.

BACKGROUND Periorbital measurements such as margin to reflex distances, palpebral fissure height, and scleral show are critical in diagnosing and managing conditions like ptosis and disorders of the eyelid. However, deployment of automated periorbital measurement algorithms in structured research workflows remains limited by the lack of integrated capture and data management infrastructure. OBJECTIVE We developed and evaluated Glorbit, a lightweight, browser-based application for automated periorbital distance measurement using artificial intelligence (AI). The objective was to evaluate end-to-end workflow feasibility of the platform under simulated, operator-run conditions. METHODS The application integrates a DeepLabV3 segmentation model into a modular image processing pipeline with secure, site-specific Google Cloud storage, supporting local preprocessing and cloud upload through Firebase-authenticated logins. The full workflow—metadata entry, facial image capture, segmentation, and upload—was tested. After the session, the participants completed a Likert-style survey. RESULTS Glorbit successfully ran on all tested platforms, including laptops, tablets, and mobile phones across major browsers. A total of 15 volunteers were enrolled in this study in which the app completed predefined workflow steps in all simulated, operator-run sessions. The segmentation model produced outputs on all images, and the average session duration was 101.7 (SD 17.5) seconds. Simulated experience scores on a 5-point Likert scale were uniformly high. CONCLUSIONS Glorbit is a cross-platform application that supports structured periorbital image capture and automated inference within a unified workflow. In simulated, operator-run testing, the platform demonstrated successful execution of predefined workflow steps across devices. These findings support the technical feasibility of the system as a research-oriented data collection framework and may inform future evaluations in broader research settings.

Background/Objectives: Marinobufagenin (MBG) is a biomarker that is found to be high in pre-eclampsia (preE), and thus is relevant in the pathogenesis of obstetric complications. MBG is thought to possibly be implicated in harmful signaling within cytotrophoblasts (CTBs) of the placenta. In this study, we evaluated how anti-MBG human monoclonal antibody can alter cellular signaling in CTBs and in a rat model of preE. Methods: CTB cell proliferation, migration, and invasion as a result of MBG, both with and without anti-MBG present, were monitored via cell-based studies. Pro-angiogenic and anti-angiogenic factors in response to MBG with and without antibody were measured. Finally, we evaluated the lead anti-MBG antibody in comparison with the parent murine antibody in a rat model of preE. Results: CTB cells exposed to ≥1 nM MBG showed decreased (p < 0.05) proliferation, migration, and invasion, decreased secretion of VEGF and PIGF, and increased secretion of sFlt-1 and sEng. Pretreatment with anti-MBG significantly (p < 0.05) attenuated MBG-induced CTB dysfunction and modulation of VEGF, PIGF, sFlt-1, and sEng expression. In the rat model, anti-MBG treatment normalized blood pressure, reduced proteinuria, and eliminated fetal effects. Conclusions: MBG is a potential causative agent for preE, as it causes dysfunction in CTBs due to anti-angiogenic milieu. Our study suggests that anti-MBG antibody binds to MBG, neutralizing it and preventing downstream signaling in vitro. In a rat model of preE, treatment with anti-MBG antibody was effective at normalizing blood pressure, kidney function, and fetal birth weights. These data suggest that a human monoclonal antibody with high specificity and affinity for MBG has potential as a therapeutic agent for preE.

Background/Objectives: Marinobufagenin (MBG), a cardiotonic steroid, is distinctly elevated in preeclampsia (preE) and directly contribute to pathogenesis, possibly through deleterious signaling in cytotrophoblasts (CTBs). In this study, we evaluated the effects of anti-MBG human monoclonal antibody on cellular signaling in CTBs and in a rat model of preE. Methods: CTB cell proliferation, migration, and invasion in response to MBG with and without anti-MBG were evaluated by cell-based studies. Pro-angiogenic and anti-angiogenic factors in response to MBG with and without antibody were measured. Finally, we evaluated the lead anti-MBG antibody in comparison with the parent murine antibody in a rat model of preE. Results: CTB cells exposed to ≥ 1 nM MBG showed decreased (p < 0.05) proliferation, migration and invasion and decreased secretion of VEGF and PIGF, and increased secretion of sFlt-1 and sEng. Pretreatment with anti-MBG significantly (p < 0.05) attenuated the MBG-induced CTBs dysfunction and modulation of VEGF, PIGF, sFlt-1, and sEng expression. In the rat model, anti-MBG treatment normalized blood pressure, reduced proteinuria, and eliminated fetal effects. Conclusions: MBG is a potential causative agent for preE as it causes dysfunction in CTBs due to anti-angiogenic milieu. Our study suggests that anti-MBG antibody binds to MBG, neutralizing it, and preventing downstream signaling in vitro. In a rat model of preE, treatment with anti-MBG antibody was effective at normalizing blood pressure, kidney function, and fetal birth weights. These data suggest that a human monoclonal antibody with high specificity and affinity for MBG has potential as a therapeutic for preE.

Sub-Saharan Africa has long been prone to widespread mosquito-borne diseases affecting both humans and animals. However, the presence and impact of West Nile virus (WNV) among livestock in Ethiopia have not been thoroughly investigated. The objective of this study was to investigate the seroprevalence of West Nile virus in livestock in the Afar region using serological methods. A total of 736 serum samples were collected from 224 cattle, 155 camels, 144 sheep, 121 goats, and 92 donkeys in the Amibara and Haruka districts of the Afar region selected using haphazard sampling. Among 736 tested livestock serum samples, 50.7% (373/736) showed anti-WNV IgG antibodies evaluated using the ID screen^® WNV competition multispecies ELISA kits (95% CI: 47–54.4%; p < 0.01). The seroprevalence was higher (p < 0.01) in donkeys (76.1%), followed by camels (69.1%), cattle (52.2%), goats (34.7%), and sheep (25.7%). The study showed a statistically significant difference of WNV seropositivity between species of animals AOR (1.5), 95% CI (1.038–2.212) (p < 0.01). Compared with sheep, donkeys had a seven-fold higher chance of being seropositive for WNV infection (OR: 6.447, 95% CI = 3.888–10.688) (p < 0.01). This study emphasizes how common WNV infection is in Ethiopia’s pastoral Afar region. It is imperative to consider consistent surveillance of WNV infection and prompt management of identified WNV disease in clinical practice. A clear need exists to build additional research capacity regarding WNV infections among both humans and animals.

The distribution, epidemiology, and socioeconomic impact of Rift Valley fever (RVF) and West Nile (WN) viruses are poorly known in areas of sub‐Saharan countries like Ethiopian pastoral region. The human and livestock density in the area has increased greatly in recent years, but little work has been done on arboviral diseases and their potential impact on human and livestock health. The aim of this study was to detect the circulation of zoonotic arboviruses such as Rift Valley fever virus and West Nile viruses in the livestock population and to estimate seroprevalence in Afar pastoral area northeast Ethiopia. Cross‐sectional serological survey was carried out in 736 serum samples from which cattle (224), camel (155), goats (121), sheep (144), and donkeys (92) were tested for the presence of anti‐RVFV and anti‐WNV IgG using a competitive enzyme‐linked immunosorbent assay (c‐ELISA) in two purposively selected districts of the Afar region. The present study revealed a 9.1% (95% CI = 8.86–9.29) seroprevalence of co‐circulation of RVF and WNV. High 32/155 (20.7%) seroprevalence of co‐circulation was seen in camels, followed by goat 14/121 (11.6%), cattle 16/224 (7.14%), and sheep 5/144 (3.5%), respectively, and higher 41/421 (9.7%) seroprevalence of co‐circulation was observed in Amibara district than Haruka district. Camels were seven times (OR: 7.016, 95% CI = 2.639–18.653) more likely to be seropositive for the co‐circulation than sheep (p ≤ 0.001). Livestock herds found in Amibara district were 1.2 times (OR: 1.165, 95% CI = 0.680–1.996) more likely to be seropositive for RVFV infection than those in Haruka areas. Given the co‐occurrence of RVFV and WNV circulations, along with often suboptimal human and animal health surveillance in many similar areas’ attention should be given. Investigation of the potential socioeconomic and health impacts of zoonotic arbovirus infections in such areas is crucial. Since both RVFV and WNFV are transmitted through a mosquito vector, avoiding mosquito bites is the primary method of prevention.

The BCR comprises a membrane-bound Ig that is noncovalently associated with a heterodimer of CD79A and CD79B. While the BCR Ig component functions to sense extracellular Ag, CD79 subunits contain cytoplasmic ITAMs that mediate intracellular propagation of BCR signals critical for B cell development, survival, and Ag-induced activation. CD79 is therefore an attractive target for Ab and chimeric Ag receptor T cell therapies for autoimmunity and B cell neoplasia. Although the mouse is an attractive model for preclinical testing, due to its well-defined immune system, an obstacle is the lack of cross-reactivity of candidate therapeutic anti-human mAbs with mouse CD79. To overcome this problem, we generated knockin mice in which the extracellular Ig-like domains of CD79A and CD79B were replaced with human equivalents. In this study, we describe the generation and characterization of mice expressing chimeric CD79 and report studies that demonstrate their utility in preclinical analysis of anti-human CD79 therapy. We demonstrate that human and mouse CD79 extracellular domains are functionally interchangeable, and that anti-human CD79 lacking Fc region effector function does not cause significant B cell depletion, but induces 1) decreased expression of plasma membrane-associated IgM and IgD, 2) uncoupling of BCR-induced tyrosine phosphorylation and calcium mobilization, and 3) increased expression of PTEN, consistent with the levels observed in anergic B cells. Finally, anti-human CD79 treatment prevents disease development in two mouse models of autoimmunity. We also present evidence that anti-human CD79 treatment may inhibit Ab secretion by terminally differentiated plasmablasts and plasma cells in vitro.

Inflammaging, the increase of proinflammatory processes with increasing age, has multiple mechanisms from increasing numbers of senescent cells secreting cytokines to changes in metabolic processes. Alterations of oxygen metabolism with aging, especially decreased levels of O ₂ with age resulting from endocrine and cardiovascular dysfunction as well as desensitization of cellular response to hypoxia, may exacerbate inflammaging, which in turn creates further oxygen metabolic dysfunction. During aging, decline in levels of erythrocyte 2,3-bisphosphoglycerate (2,3-BPG), BPG mutase, and adenosine A2B receptor, a key adenosine signaling receptor that can augment 2,3-BPG expression, may fail to protect sensitive brain tissue from subtly reduced O ₂ levels, in turn resulting in increased numbers of activated microglia and secretion of proinflammatory cytokines, ultimately promoting inflammaging and senescence of endothelial cells. Interventions to restore O ₂ levels directly or via increasing 2,3-BPG may help promote cognitive health in old age, but significant work to quantify the degree of reduced O ₂ during aging in mammals, and especially humans, needs to be pursued.

Intermittent fasting (IF) is the practice of restricting food intake for 12–48 hours per fasting cycle over a prolonged period of time. Previous study shows beneficial health effects such as weight loss and lower risk for cardiometabolic diseases. Although reduced calorie intake may account for some of the observed benefits of IF, exact mechanisms are still unclear. Recent evidence indicates that IF may lead to remodeling and increased taxonomic diversity in the human gut microbiome. In particular, the Lachnospiraceae family of anaerobic bacteria increased during fasting. This family, in the order Clostridiales, promotes butryogenesis in the gut, a process that is associated with healthful metabolic and prolongevity effects. IF-associated alterations to the microbiome may play a key role in the metabolic and potential healthspan-enhancing benefits of IF and dietary restriction.