Reciprocal Activation Research Articles

Unveiling the Underlying Motor Control Mechanism of Arm-trunk-leg Coordinated Humanoid Exoskeletal Interlimb Locomotor Robotic Neurorehabilitation

Background Arm-leg coordinated humanoid exoskeletal robots offer a solution to the ‘unnatural’ gait patterns seen in current leg-only exoskeletons, but their neuromechanical control mechanisms require further exploration. Objective This study investigates the neuromechanical properties and modulation of cervical and lumbar central pattern generators (CPGs) across different gait training conditions. Methods Eleven healthy adults (mean age: 25.27 ± 1.95 years; 5 females, 6 males) participated in gait analysis involving kinematics, kinetics, and electromyography during treadmill-based arm-leg interlimb coordination (TIT), robot-assisted arm-leg coordination (RIT), and robot-assisted leg-only coordination (RLT). Results Trunk counterrotation and shoulder, hip, and knee kinematics in RIT closely resembled those in TIT (5–10% variance) but significantly differed from RLT (30–40% variance). Electromyography revealed normalized reciprocal activation between the anterior and posterior deltoids (r = -0.48), tibialis anterior, and gastrocnemius (r = -0.68) during RIT, while RLT showed altered agonist-antagonist co-activation (r = 0.84). Conclusions These findings highlight the importance of cervical and lumbar CPG interaction in natural locomotion, suggesting that arm-leg coordinated exoskeletons could improve neurorehabilitation for individuals recovering from stroke or spinal cord injury.

Low Parental Empathy Predicts Callous‐Unemotional Traits in Children With Enhanced Autonomic Sensitivity to the Environment

ABSTRACTIntroductionRecent evidence suggests that autonomic nervous system functioning can reflect individual differences in sensitivity to the environment, which in turn moderates the effects of family context on psychopathic and antisocial behaviour. Although some preliminary research suggests that the coordination of the sympathetic (SNS) and parasympathetic (PNS) branches of the autonomic nervous system may reflect sensitivity to environmental influences, the majority of research to date has been limited in its focus on just one branch, and little is known about the effects of parental empathy on a child's psychopathic‐like traits. This study aimed to address this gap by examining if the SNS and PNS jointly moderate the prospective contribution of parental empathy to youth psychopathic‐like traits.MethodsMale and female children (n = 340; age = 8–10 years) from the community completed an emotion regulation task during which their skin conductance level (SCL) and respiratory sinus arrhythmia (RSA) activity were recorded. Parental cognitive, affective, and somatic empathy were reported by the caregivers. The child's psychopathic‐like traits were reported by the caregiver and the child and were reassessed 1 year later.ResultsHierarchical regression indicated that after controlling for child sex, age, and concurrent CU traits, low affective empathy in parents predicted higher CU traits 1 year later in youths who exhibited reciprocal sympathetic activation (i.e., SNS activation along with PNS withdrawal), reflecting their enhanced sensitivity to the environment. No such effects were found for other psychopathic‐like traits.ConclusionsFindings suggest that reciprocal SNS reactivity may be a biological indicator for sensitivity to environmental influence and highlight the importance of examining the interactions among multiple systems to better understand the aetiology of psychopathic traits.

Pengaruh Kompetensi dan Komunikasi Terhadap Kinerja Karyawan PT Beta Pharmacon

Performance is the work result that can be achieved by individuals or work teams within an organization to achieve organizational goals within a certain period of time. Competence is defined as an individual's ability to carry out a job correctly and with excellence. Reciprocal activities in conveying messages, using message delivery media or without media, are defined as communication processes. This research aims to determine and analyze the influence of competency and communication variables on the performance of PT Beta Pharmacon. The quantitative descriptive research design used multiple linear regression verification a population of 388 employees, and used a proportional starting random sampling technique with the Slovin formula and a sample of 80 was determined. The results of the descriptive analysis of the competency are in good criteria, the communication is in good criteria and the performance is in good criteria. The results of the verification analysis of the competency variable (X1) partially have a very significant influence on employee performance, the communication variable (X2) partially has a very significant influence on employee performance, while the competency variable (X1) and communication variable (X2) have a simultaneous influence on employee performance (Y).

Using Theory and Technology to Build an Interprofessional Community of Human Trafficking Educators.

Human trafficking, a global health crisis, requires interprofessional responses. Skilled teachers are needed to train health care providers about human trafficking. To promote nonhierarchical interprofessional collaborative learning, we applied social cognitive and experiential learning theories within a dialectical constructivist program design to scaffold participants' knowledge and leveraged technology to build and sustain the program's community. WhatsApp and Flipgrid connected participants and faculty prior to the program. Participants' reflections on experiences were used to inform confidential and respectful information sharing. Live case presentations were interwoven with prerecorded didactics, Zoom break-out case analyses, and Q&A sessions with trafficking survivors. Participants used learning theories to cocreate and teach about labor and sex trafficking, disclosure, and the law. A reciprocal teaching activity facilitated participants' integration of new knowledge with authentic work responsibilities. Constructive peer feedback on the content, clarity, and engagement of their teaching reinforced participants' self-efficacy in expanding their education work in their home organizations. As of 2021, 156 physicians, nurses, social workers, advanced practice providers, psychologists, and public health workers, from the United States, United Kingdom, Canada, and Trinidad/Tobago, have graduated from the program. Three-month postprogram surveys indicated lasting knowledge and skills changes in use of the Stop, Observe, Ask, Refer framework, teaching with adult learning principles, and creating organizational trafficking protocols. The strategic application of learning theory and technology has enabled us to foster a nonhierarchical community of interprofessional learners, cultivating a dynamic network of educators who continue to make international impacts on people with an experience of human trafficking.

Abstract A001: Reciprocal activation of breast cancer metastases and the lung epithelium during metastatic outgrowth

Abstract Introduction. During metastasis, tumor cells interact extensively with endogenous cells in distant organs. These interactions change the behavior of these surrounding cell populations, often inducing a more pro-tumor microenvironment. The lung is one of the most common sites of breast cancer (BC) metastasis, and in the lung, alveolar epithelial cells are the most common cell type. My data suggest that the lung alveolar epithelium contributes to metastatic outgrowth, and I hypothesize that BC lung micrometastases activate surrounding lung epithelial cells which, in turn, support the outgrowth of BC metastases within the lung. The overall purpose of these studies is to identify factors secreted by resident lung epithelial cells that could be used as metastasis-specific therapeutic targets and/or agents. Methods. To test this hypothesis, I utilized immunocompetent preclinical BC metastasis models to study lung metastatic outgrowth, the stage at which most patients are diagnosed with metastatic disease. A custom imaging panel was developed to quantify lung wound repair, and single-cell RNA-sequencing (scRNAseq) was performed on mouse lungs with high or low metastatic burden to identify genes in the lung epithelium that produce potentially targetable pro-metastatic secreted factors. No-contact co-culture experiments were used to directly study reciprocal paracrine interactions between lung type II alveolar epithelial (AT2) and BC cells. Results. A wound repair-related phenotype, characterized by chronic inflammation, developed within the lung microenvironment during metastatic outgrowth, including an increase in the number and activation of AT2 cells surrounding metastases as they grow. Single-cell RNA-sequencing of mouse lungs with a high vs. low metastatic burden showed that metastatic outgrowth significantly changed AT2 gene expression resulting in a modified secretome. No-contact co-culture experiments indicated that TNBC cells directly alter AT2 gene expression, while AT2 secreted factors promoted TNBC growth. I investigated the possible mechanism(s) responsible for these effects and discovered that AT2 pulmonary surfactant protein and lipid levels are altered by BC-derived secreted factors. Interestingly, treatment with the naturally derived calf surfactant Infasurf, which contains native surfactant proteins/lipids and is FDA-approved for use in premature infants, inhibited BC proliferation. Conclusion. Low levels of surfactant are commonly associated with pulmonary disease and lung cancer. My BC lung metastasis data suggest that something similar may be occurring in the lungs during metastatic outgrowth. Overall, my studies demonstrate the potential for targeting lung epithelial cells in the metastatic microenvironment, in addition to directly targeting malignant cells, as an effective way to treat and manage BC lung metastases. Citation Format: Jessica L. Christenson, Nicole S. Spoelstra, Jennifer K. Richer. Reciprocal activation of breast cancer metastases and the lung epithelium during metastatic outgrowth [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr A001.

Antithrombotic coating with sheltered positive charges prevents contact activation by controlling factor XII-biointerface binding.

Antithrombotic surfaces that prevent coagulation activation without interfering with haemostasis are required for blood-contacting devices. Such materials would restrain device-induced thrombogenesis and decrease the need for anticoagulant use, thereby reducing unwanted bleeding. Here, by optimizing the interactions with coagulation factor XII rather than preventing its surface adsorption, we develop a substrate-independent antithrombotic polymeric coating with sheltered positive charges. The antithrombic properties of the coating were demonstrated in vitro with human blood and in vivo using a carotid artery-jugular vein shunt model in rabbits. The coating exhibits a strong interaction with factor XII, but results in a low reciprocal activation of the contact pathway that triggers clot formation. These findings contradict the prevailing strategy of designing antithrombotic materials through protein-repelling surfaces. Overall, the polymeric coating we describe can benefit most blood-contacting devices and is a useful engineering guideline for designing surfaces with improved antithrombotic properties.

Dynamic reciprocal interactions between activated T cells and tumor associated macrophages drive macrophage reprogramming and proinflammatory T cell migration within prostate tumor models

Tumor-associated macrophages (TAMs) have been implicated as a tumor microenvironment (TME) cell population, which may be playing a vital role in the inhibition of effective T cell responses in the prostate TME. In this manuscript, we leverage a novel microscale cell culture platform, known as Stacks, to investigate mono-, co-, and tri-culture TME models comprised of prostate tumor cell lines, primary macrophages, and autologous T cells from patients with prostate cancer. Through multiplexed analysis of these multi-cellular prostate tumor models, we capture a dynamic interaction between primary TAMs and activated T cells that resulted in reciprocal proinflammatory activation of both cell populations upon interaction. These findings suggest that activated T cells are capable of reprogramming immunosuppressive TAMs in the context of prostate tumor models and that TAM reprogramming may play a key supportive role in restoring proinflammatory T cell tumor responses in the prostate TME.

An autocatalytic hybridization circuit-based FRET aptasensor for detection of low-abundant sulfameter in human serum

Exposure to antibiotics is considered a potential risk factor for human health. Yet, the extensive and cost-effective detection of low-abundant antibiotics in complex matrices remains a significant challenge. Herein, an aptamer and an autocatalytic hybridization circuit (AHC) were used to fabricate a fluorescence resonance energy transfer (FRET) platform to detect sulfameter (SME) in human serum. The AHC system comprised two mutually motivated hybridization chain reactions (HCR) modules, ultimately producing long-branched DNA copolymeric nanowires. This mutually reciprocal activation of two HCR modules enables continuous signal amplification, providing the AHC system with wide linear range and high sensitivity for the SME detection. Compared to the HCR-based aptasensor, the AHC-based aptasensor exhibited a wider linear range and improved sensitivity (3.3 times greater). Under optimal conditions, the fluorescent AHC-based aptasensor demonstrated a linear range (R2 was 0.996) from 0.5 to 2000 nM, with a low detection limit of 0.301 nM (S/N = 3). The fluorescent aptasensor was also validated by SME-spiked human serum samples, showing average recoveries ranging from 96.40 % to 109.30 %, with a relative standard deviation below 10.45 %. Furthermore, when tested on six human serum samples, the aptasensor results were consistent with those obtained from the commercial ELISA method. These findings demonstrate that the proposed aptasensor provides a promising approach for the practical monitoring of low-abundant SME in human serum.

The Role of Social Capital on Consumer Behavior and Sustainability of Religious Business Units (Case Study of Waringin Store, Mataram City)

This research is aim to get a analyze social capital's impact on the sustainability of a business unit in the trade sector for ceremonial or religious activity products for Hindu communities. The data used is primary data obtained through in-depth interviews with a data triangulation method. The findings of the research indicate that there is a relationship between social capital and business sustainability, through Bourdieu's theory on how social capital can generate economic resources. The expression of socio-economic relationships is marked by activities in behaviors or actions that support social interaction, such as partnerships, activities characterized by networking participation, reciprocity (exchange or reciprocal activities between individuals or groups), trust, values, social norms, and proactive actions. The application of social capital in the case of Toko Waringin facilitates customers through access to a very strategic location, availability of products, and a variety of Hindu religious ceremonial supplies, with a good inventory system and flexible service times.

Self-assembled PROTACs enable protein degradation to reprogram the tumor microenvironment for synergistically enhanced colorectal cancer immunotherapy

Both β-catenin and STAT3 drive colorectal cancer (CRC) growth, progression, and immune evasion, and their co-overexpression is strongly associated with a poor prognosis. However, current small molecule inhibitors have limited efficacy due to the reciprocal feedback activation between STAT3 and β-catenin. Inspired by the PROteolysis TArgeting Chimera (PROTAC), a promising pharmacological modality for the selective degradation of proteins, we developed a strategy of nanoengineered peptide PROTACs (NP-PROTACs) to degrade both β-catenin and STAT3 effectively. The NP-PROTACs were engineered by coupling the peptide PROTACs with DSPE-PEG via disulfide bonds and self-assembled into nanoparticles. Notably, the dual degradation of β-catenin and STAT3 mediated by NP-PROTACs led to a synergistic antitumor effect compared to single-target treatment. Moreover, NP-PROTACs treatment enhanced CD103+ dendritic cell infiltration and T-cell cytotoxicity, alleviating the immunosuppressive microenvironment induced by β-catenin/STAT3 in CRC. These results highlight the potential of NP-PROTACs in facilitating the simultaneous degradation of two pathogenic proteins, thereby providing a novel avenue for cancer therapy.

Single session of direction-specific training using auditory cues improves anticipatory postural adjustments to lateral perturbations.

When exposed to a predictable external perturbation, humans typically generate anticipatory postural adjustments (APAs) to minimize potential body disturbance. After a single session of training, individuals demonstrated the ability to rely solely on an auditory cue to elicit appropriate APAs in response to an external postural perturbation. However, whether the generation of APAs requires directional specific training remains unclear. The aim of this study was to assess whether directional-specific training with auditory cues is necessary for the generation of appropriate APA responses. Ten young adults were exposed to external perturbations targeting either their left or right shoulders, with or without an auditory cue prior to the physical impact. Electromyography (EMG) activities of sixteen trunk and leg muscles and center-of-pressure (COP) displacements were recorded and analyzed during the anticipatory and compensatory phases of postural control. Outcome measures included the latencies and integrals of muscle activities, COP displacements, and indices of co-contraction and reciprocal activation of muscles. The results revealed that, after training with right-side perturbations accompanied by an auditory cue, young adults exhibited earlier and more efficient APA responses to right-side perturbations relying only on the auditory cue. Additionally, they displayed earlier APA responses in some muscles to left-side perturbations, although these responses were less efficient. Our findings suggest that young adults could generate effective APAs to external perturbations relying on an auditory cue after a single training session; however, these responses were directional specific.

HOTAIR Promotes the Hyperactivation of PI3K/Akt and Wnt/β-Catenin Signaling Pathways via PTEN Hypermethylation in Cervical Cancer.

The mechanisms underlying the sustained activation of the PI3K/AKT and Wnt/β-catenin pathways mediated by HOTAIR in cervical cancer (CC) have not been extensively described. To address this knowledge gap in the literature, we explored the interactions between these pathways by driving HOTAIR expression levels in HeLa cells. Our findings reveal that HOTAIR is a key regulator in sustaining the activation of both signaling pathways. Specifically, altering HOTAIR expression-either by knockdown or overexpression-significantly influenced the transcriptional activity of the PI3K/AKT and Wnt/β-catenin pathways. Additionally, we discovered that HIF1α directly induces HOTAIR transcription, which in turn leads to the epigenetic silencing of the PTEN promoter via DNMT1. This process leads to the sustained activation of both pathways, highlighting a novel regulatory axis involving HOTAIR and HIF1α in cervical cancer. Our results suggest a new model in which HOTAIR sustains reciprocal activation of the PI3K/AKT and Wnt/β-catenin pathways through the HOTAIR/HIF1α axis, thereby contributing to the oncogenic phenotype of cervical cancer.

The roles of collapsing contexts and TikTok’s features in reciprocal trolling

ABSTRACT The collapsing of social contexts online and its impact on identify, privacy, and self-presentation have been widely documented. However, the increased levels of conflict and misunderstanding that those collapsing contexts trigger requires more scholarly attention. Informed by social informatics, this paper examines collapsing contexts and TikTok’s features that support reciprocal trolling. A comparative case analysis of three TikTok influencers reveals how they took advantage of the platform’s features (Duet, Stich, Share, and Sounds); how they developed their own trolling style and utilized a different repertoire of trolling tactics in initiating and in responding to trolling; and how they proactively utilized social, spatial, and temporal collapsing contexts in their reciprocal trolling activities. The study makes two significant contributions to research by (1) extending our understanding of collapsing contexts beyond the reactive notion of flattened contexts, contributing to social informatics a nuanced understanding of context; and by (2) demonstrating how TikTok’s features foster reciprocal trolling and the process by which it spills over into other social media platforms, raising a need for more cross-platform research.

Sciatic nerve stimulation alleviates neuropathic pain and associated neuroinflammation in the dorsal root ganglia in a rodent model

BackgroundSatellite glial cells (SGCs) in the dorsal root ganglia (DRG) play a pivotal role in the formation of neuropathic pain (NP). Sciatic nerve stimulation (SNS) neuromodulation was reported to alleviate NP and reduce neuroinflammation. However, the mechanisms underlying SNS in the DRG remain unclear. This study aimed to elucidate the mechanism of electric stimulation in reducing NP, focusing on the DRG.MethodsL5 nerve root ligation (NRL) NP rat model was studied. Ipsilateral SNS performed 1 day after NRL. Behavioral tests were performed to assess pain phenotypes. NanoString Ncounter technology was used to explore the differentially expressed genes and cellular pathways. Activated SGCs were characterized in vivo and in vitro. The histochemical alterations of SGCs, macrophages, and neurons in DRG were examined in vivo on post-injury day 8.ResultsNRL induced NP behaviors including decreased pain threshold and latency on von Frey and Hargreaves tests. We found that following nerve injury, SGCs were hyperactivated, neurotoxic and had increased expression of NP-related ion channels including TRPA1, Cx43, and SGC-neuron gap junctions. Mechanistically, nerve injury induced reciprocal activation of SGCs and M1 macrophages via cytokines including IL-6, CCL3, and TNF-α mediated by the HIF-1α-NF-κB pathways. SNS suppressed SGC hyperactivation, reduced the expression of NP-related ion channels, and induced M2 macrophage polarization, thereby alleviating NP and associated neuroinflammation in the DRG.ConclusionsNRL induced hyperactivation of SGCs, which had increased expression of NP-related ion channels. Reciprocal activation of SGCs and M1 macrophages surrounding the primary sensory neurons was mediated by the HIF-1α and NF-κB pathways. SNS suppressed SGC hyperactivation and skewed M1 macrophage towards M2. Our findings establish SGC activation as a crucial pathomechanism in the gliopathic alterations in NP, which can be modulated by SNS neuromodulation.

유아교사의 사회적 초인지 역량 함양을 위한 상보적 교육모형 및 프로그램 개발

Purpose: The purpose of this study was to develop a reciprocal education model and program to cultivate the social metacognitive competency of early childhood teachers. Methods: This educational model confirmed the validity of the contents of experts, and the appropriateness of the educational program was confirmed through pilot tests. Results: The procedure of the reciprocal education model consisted of Aware-Analysis and Monitoring (AM), Planning and Monitoring (PM), Reciprocal teaching Activities (RA), and Reflection and Evaluation (RE). The program was developed so that early childhood teachers could acquire and apply metacognitive strategies. Conclusion: This study will be helpful for cultivating professionalism of early childhood teachers.

The First Reciprocal Activities of Chiral Peptide Pharmaceuticals: Thymogen and Thymodepressin, as Examples.

Peptides show high promise in the targeting and intracellular delivery of next-generation biotherapeutics. The main limitation is peptides' susceptibility to proteolysis in biological systems. Numerous strategies have been developed to overcome this challenge by chemically enhancing the resistance to proteolysis. In nature, amino acids, except glycine, are found in L- and D-enantiomers. The change from one form to the other will change the primary structure of polypeptides and proteins and may affect their function and biological activity. Given the inherent chiral nature of biological systems and their high enantiomeric selectivity, there is rising interest in manipulating the chirality of polypeptides to enhance their biomolecular interactions. In this review, we discuss the first examples of up-and-down homeostasis regulation by two enantiomeric drugs: immunostimulant Thymogen (L-Glu-L-Trp) and immunosuppressor Thymodepressin (D-Glu(D-Trp)). This study shows the perspective of exploring chirality to remove the chiral wall between L- and D-biomolecules. The selected clinical result will be discussed.

AS1411 binds to nucleolin via its parallel structure and disrupts the exos-miRNA-27a-mediated reciprocal activation loop between glioma and astrocytes

The interaction between glioma cells and astrocytes promotes the proliferation of gliomas. Micro-RNAs (miRNAs) carried by astrocyte exosomes (exos) may be involved in this process, but the mechanism remains unclear. The oligonucleotide AS1411, which consists of 26 bases and has a G-quadruplex structure, is an aptamer that targets nucleolin. In this study, we demonstrate exosome-miRNA-27a-mediated cross-activation between astrocytes and glioblastoma and show that AS1411 reduces astrocytes' pro-glioma activity. The enhanced affinity of AS1411 toward nucleolin is attributed to its G-quadruplex structure. After binding to nucleolin, AS1411 inhibits the entry of the NF-κB pathway transcription factor P65 into the nucleus, then downregulates the expression of miRNA-27a in astrocytes surrounding gliomas. Then, AS1411 downregulates astrocyte exosome-miRNA-27a and upregulates the expression of INPP4B, the target gene of miRNA-27a in gliomas, thereby inhibiting the PI3K/AKT pathway and inhibiting glioma proliferation. These results were verified in mouse orthotopic glioma xenografts and human glioma samples. In conclusion, the parallel structure of AS1411 allows it to bind to nucleolin and disrupt the exosome-miRNA-27a-mediated reciprocal activation loop between glioma cells and astrocytes. Our results may help in the development of a novel approach to therapeutic modulation of the glioma microenvironment.

Is autonomic functioning distinctly associated with anxiety and unsociability in preschoolers?

There are many benefits of peer interactions for children's social, emotional, and cognitive development, and isolation from peers may have negative consequences for children. Although biological processes may underlie social withdrawal broadly, distinct patterns may be associated with withdrawal behaviors depending on their underlying motivation (e.g., shy versus disinterested). This study investigated the role of autonomic nervous system activity, as assessed via skin conductance level (SCL) and respiratory sinus arrhythmia (RSA) in predicting changes in unsociability (e.g., lack of interest in peers) and anxious-fearfulness (e.g., discomfort among peers). Data were collected using a community sample of 92 US preschool children (45.7% female; Mage = 45.51 months, SDage = 3.81 months) at two time points one year apart. Gender differences were also explored. Baseline physiology was assessed while viewing a neutral video clip, and reactivity was assessed while viewing social exclusion and post-aggression discussion videos. For all children, coinhibition (i.e., SCL inhibition accompanied by RSA inhibition) to the post-aggression discussion video and blunted SCL activation to the exclusion video were prospectively associated with higher levels of anxious-fearfulness one year later. For boys only, baseline reciprocal sympathetic activation (i.e., SCL activation and RSA inhibition) was prospectively related to higher levels of unsociability one year later. For girls only, RSA inhibition in response to the post-aggression discussion video was prospectively related to higher levels of unsociability one year later. Findings contribute to a growing literature on autonomic reactivity in preschoolers' adjustment and suggest possible differences in the physiological processes underlying unsociability and anxious-fearfulness.

Wnt5a/β-catenin-mediated epithelial-mesenchymal transition: a key driver of subretinal fibrosis in neovascular age-related macular degeneration

BackgroundNeovascular age-related macular degeneration (nAMD), accounts for up to 90% of AMD-associated vision loss, ultimately resulting in the formation of fibrotic scar in the macular region. The pathogenesis of subretinal fibrosis in nAMD involves the process of epithelial–mesenchymal transition (EMT) occurring in retinal pigment epithelium (RPE). Here, we aim to investigate the underlying mechanisms involved in the Wnt signaling during the EMT of RPE cells and in the pathological process of subretinal fibrosis secondary to nAMD.MethodsIn vivo, the induction of subretinal fibrosis was performed in male C57BL/6J mice through laser photocoagulation. Either FH535 (a β-catenin inhibitor) or Box5 (a Wnt5a inhibitor) was intravitreally administered on the same day or 14 days following laser induction. The RPE-Bruch's membrane-choriocapillaris complex (RBCC) tissues were collected and subjected to Western blot analysis and immunofluorescence to examine fibrovascular and Wnt-related markers. In vitro, transforming growth factor beta 1 (TGFβ1)-treated ARPE-19 cells were co-incubated with or without FH535, Foxy-5 (a Wnt5a-mimicking peptide), Box5, or Wnt5a shRNA, respectively. The changes in EMT- and Wnt-related signaling molecules, as well as cell functions were assessed using qRT-PCR, nuclear-cytoplasmic fractionation assay, Western blot, immunofluorescence, scratch assay or transwell migration assay. The cell viability of ARPE-19 cells was determined using Cell Counting Kit (CCK)-8.ResultsThe in vivo analysis demonstrated Wnt5a/ROR1, but not Wnt3a, was upregulated in the RBCCs of the laser-induced CNV mice compared to the normal control group. Intravitreal injection of FH535 effectively reduced Wnt5a protein expression. Both FH535 and Box5 effectively attenuated subretinal fibrosis and EMT, as well as the activation of β-catenin in laser-induced CNV mice, as evidenced by the significant reduction in areas positive for fibronectin, alpha-smooth muscle actin (α-SMA), collagen I, and active β-catenin labeling. In vitro, Wnt5a/ROR1, active β-catenin, and some other Wnt signaling molecules were upregulated in the TGFβ1-induced EMT cell model using ARPE-19 cells. Co-treatment with FH535, Box5, or Wnt5a shRNA markedly suppressed the activation of Wnt5a, nuclear translocation of active β-catenin, as well as the EMT in TGFβ1-treated ARPE-19 cells. Conversely, treatment with Foxy-5 independently resulted in the activation of abovementioned molecules and subsequent induction of EMT in ARPE-19 cells.ConclusionsOur study reveals a reciprocal activation between Wnt5a and β-catenin to mediate EMT as a pivotal driver of subretinal fibrosis in nAMD. This positive feedback loop provides valuable insights into potential therapeutic strategies to treat subretinal fibrosis in nAMD patients.

Effect of Hindlimb Unloading on Hamstring Muscle Activity in Rats

Introduction: The changes in knee axial rotation play an important role in traumatic and non-traumatic knee disorders. It is known that support afferentation can affect the axial rotator muscles. The condition of innervation of the semitendinosus (ST) and biceps femoris posterior (BFp) has changed in non-terrestrial and terrestrial vertebrates in evolution; thus, we hypothesized this situation might be replayed by hindlimb unloading (HU). Methods: In the present study, the EMG activity of two hamstring muscles, m. ST and m. BFp, which are antagonists in axial rotation of the tibia, was examined before and after 7 days of HU. Results: During locomotion and swimming, the ST flexor burst activity increased in the stance-to-swing transition and in the retraction-protraction transition, respectively, while that of BFp remained unchanged. Both ST and BFp non-burst extensor activity increased during stepping and decreased during swimming. Conclusions: Our results show that (1) the flexor burst activity of ST and BFp depends differently on the load-dependent sensory input in the step cycle; (2) shift of the activity gradient towards ST in the stance-to-swing transition could produce excessive internal tibia torque, which can be used as an experimental model of non-traumatic musculoskeletal disorders; and (3) the mechanisms of activity of ST and BFp may be based on reciprocal activity of homologous muscles in primary tetrapodomorph and depend on the increased role of supraspinal control.