Autonomous Signaling Research Articles

Effect of Signal Design of Autonomous Vehicle Intention Presentation on Pedestrians' Cognition.

In this study, a method is devised that allows the intentions of autonomous vehicles to be effectively communicated to pedestrians and passengers via an efficient interactive interface. Visual and auditory factors are used as variables to investigate the effects of different autonomous vehicle signal factors on the judgment of pedestrians and to determine the main factors such that the best combination can be proposed. Two visual dimensions (i.e., color and flashing) and three auditory dimensions (i.e., rhythm, frequency, and melody) are used as the experimental signal variables. In addition, deceleration and waiting-to-restart scenarios are investigated. Multiple-choice questions and a subjective cognition scale are used for evaluation. The results show that the combination of green and slow rhythm can be used for the road-user-first case, whereas the combination of red and fast rhythm can be used for the vehicle-first case. Under the same intention, factors of color, flashing, rhythm, and melody are highly similar in terms of the combination mode, except for the frequency. In the deceleration and waiting-to-restart scenarios, the frequencies of the best signal are high and low frequencies, respectively. The results of this study can be used as a reference for the signal design of autonomous vehicles in the future and provide ideas for the interactions between autonomous vehicles and pedestrians.

Antigen-Independent, Autonomous B-Cell Receptor Signaling in Diffuse Large B-Cell Lymphoma

Gene expression profiling of diffuse large B-cell lymphoma (DLBCL) defines the Cell-of-origin (CoO) classification with two major subtypes, germinal center B-cell (GCB) and the more aggressive activated B-cell (ABC) type. Chronic activation of B-cell receptor (BCR) signaling with constitutive activation of the NF-κB pathway is an essential oncogenic mechanism in ABC DLBCL. BCR pathway activation with increased NF-κB activation can be caused by gain-of-function mutations of the BCR signaling cascade. We investigated whether antigen-independent, i.e. autonomous BCR signaling, akin to its essential pathogenetic role in chronic lymphocytic leukemia, acts as a functional driver mechanism in DLBCL without such gain-of-function mutations. DNA and RNA were extracted from ABC DLBCL cell lines TMD8 and OCI-Ly3 and from fresh-frozen biopsies of 18 histologically confirmed DLBCL cases. Functional BCR transcripts were identified by ARTISAN PCR and transduced into murine triple knock out (TKO) cells. TKO cells lack the recombination activation gene 2 (Rag2) and lambda 5 gene (λ5) and are therefore incapable of expressing an endogenous BCR. In addition, the BCR signal transduction molecule SLP65 is replaced with a tamoxifen-inducible estrogen receptor (ERT2) - SLP65 fusion gene. Upon transduction of both heavy and light BCR chains and staining with Indo-1, background calcium flux can be measured as Indo-1 signal intensity ratio (SIR) by flow cytometry. After restoring functionality of the BCR signaling cascade by tamoxifen, antigen-independent BCR signaling is measured as SIR, followed by maximum BCR stimulation with an isotype-specific crosslinking antibody. Autonomous BCR signaling strength (SSBCR) is quantified by an arithmetic product based on the relative Indo-1 SIR and the relative number of cells with SIR above background in the presence of tamoxifen and calibrated on crosslinking-induced maximum SIR. According to the primary hypothesis, the BCR of CARD11 wild-type, IgM-expressing TMD8 cells demonstrated strong autonomous BCR signaling (SSBCR=0.243), whereas the BCR of CARD11 mutated, IgG-expressing OCI-Ly3 cells completely lacked autonomous signaling (SSBCR=0.022). BCR from 13 primary DLBCL signaled autonomously (SSBCR=0.125-0.619), where 5 samples were negative (SSBCR=0.004-0.023) Autonomous BCR signaling was absolutely dependent on IgM isotype (Fisher's exact test: p=0.0016). CoO classification by digital gene expression (Nanostring) analyses or immunohistochemistry demonstrated autonomous BCR signaling in 8/9 ABC (non-GCB) DLBCL, 2/6 GCB DLBCL, and 2 unclassifiable DLBCL, and a transformed follicular lymphoma DLBCL. Independent of CoO, DLBCL can be assigned to 6-7 clusters based on their genetic aberrations (Chapuy 2018, Wright 2020). Fifteen DLBCL were suited for WES and targeted panel sequencing. Four cases were assigned by a probabilistic calculation to Chapuy cluster C5, 2 cases to C2, and one case to C1, while 8 cases remained unclassifiable. Clustering by the LymphGen tool assigned 3 cases to cluster ST2, one case each into MCD, EZB, and N1, 4 cases into various clusters, and 5 cases unclassified. DLBCL with autonomous BCR signaling fell into clusters C1/unclassified, C2/unclassified, C5/unclassified (n=3), unclassified/EZB, unclassified/ST2, unclassified/N1 and unclassified/unclassified (n=2). Cases lacking autonomous BCR signaling were assigned to unclassified/ST2 (n=2), C5/MCD, and unclassified/unclassified. Eleven of 13 cases with an autonomous signaling BCR lacked activating CARD11 mutations. In conclusion, autonomous BCR signaling is a potential immunological oncogenic driver in DLBCL that cannot be detected by sequencing studies, is independent of external antigen stimulation, and originates from a self-organizing conformation of the surface BCR (Minici 2017). Autonomous BCR signaling is present predominantly in ABC DLBCL and dependent on IgM isotype. Autonomous BCR signaling offers a convincing explanation for oncogenic NF-κB activation in DLBCL. Limited data suggest that the majority of DLBCL with autonomous BCR signaling fall into the ABC-dominated genetic clusters C1/C5, and MCD/N1, respectively. In summary, genetic aberrations, BCR function, and gene expression profile form 3 independent layers of DLBCL pathophysiology that need to be analyzed separately and offer independent targets for therapy.

Autonomous BCR Signaling and Genetic Aberrations in CLL-Phenotype Monoclonal B Lymphocytosis in Siblings of CLL Patients

Clonal expansion of CD5-expressing B cells, termed monoclonal B lymphocytosis (MBL), is a precursor condition for chronic lymphocytic leukemia (CLL) with an increased incidence in CLL siblings. In app. 1% of MBL carriers, MBLprogresses eventually to clinically overt CLL. The mechanisms driving benign MBL expansion and subsequent malignant progression are unknown. Autonomous signaling of the clonotypic BCR of CLL cells, i.e. signaling without stimulation by cognate antigen, is an indispensable oncogenic signal in CLL. In order to investigate the early stages in CLL ontogeny, we sought to clarify whether MBL cells express BCR with autonomous signaling capacity and to compare the prevalence of inherited risk loci and acquired genetic aberrations in MBL-CLL sibling pairs. Peripheral blood of 191 siblings of CLL patients was screened with informed consent by six-color flow cytometry. A discrete CD19+CD5+CD20lowCD79low CLL-phenotype population of 0.2 to 1863 cells / µL blood was detected in 34 siblings (17.8%) of 26 CLL patients; 32 siblings (94%) had <500 clonal CLL-phenotype cells / µL. Clonal BCR were identified from highly purified MBL cells of 17 CLL siblings by ARTISAN PCR. Five MBL clones could be assigned to CLL stereotyped subsets, including two MBL BCR belonging to CLL subset #2 with expression of the IGLV3-21 gene with a G110R mutation and one IgG-expressing MBL belonging to CLL subset #4. Eleven MBL BCR, including all 5 assigned to a CLL stereotyped subset, were transduced into murine TKO cells arrested at the pro-B-cell stage by genetic deficiency of rag2, lambda5, and slp65. Slp65 function is reconstituted by a slp65-ERT2 fusion gene that enables BCR signaling in the presence of 4-hydroxytamoxifen. Calcium mobilization was measured in Indo-1 AM-loaded, live-gated TKO cells by flow cytometry as the ratio of signal intensities (SIR) at 485 and 405 nm prior and after tamoxifen supplementation and after BCR crosslinking with isotype-specific antibody. Robust calcium mobilization was observed in all MBL BCR-transduced TKO cells in the absence of external antigen and without experimental cross-linking, regardless of assignment to a CLL subtype. Calibrated quantification of this autonomous BCR signal, however, revealed lesser intensity in MBL BCR compared to the BCR of their CLL sibling (p=0.015, Wilcoxon's matched-pair signed rank test). BCR signaling strength was correlated to the degree of MBL clonal expansion (p=0.034). According to genotyping by single nucleotide polymorphism array, whole exome sequencing (WES), and targeted panel sequencing, a polygenic CLL risk score (PRS) of 24 risk loci was higher in both CLL and MBL siblings than in reference population data, but we found no evidence for a higher PRS in CLL cases in a matched-pair comparison to their MBL siblings. With respect to acquired genetic changes, quantitative analysis of SNP array data revealed recurrent CLL-associated copy number variations (CNV) at clonal levels in all of 11 informative CLL cases. In contrast, individual CNV were present in only a minor fraction of the highly purified MBL cells in six of ten samples (p=0.003; Fisher's exact test). Likewise, variant allele frequencies (VAF) of non-shared variants as detected by WES was significantly lower in MBL samples than in CLL cases (p<0.0001). In particular, subclonal variants defined by a VAF of 0.1-0.33 were more prevalent in MBL compared to CLL siblings (p=0.014), whereas clonal variants (VAF 0.43-0.55) were equally abundant (p=0.65) in both conditions. In this comprehensive comparison of MBL and CLL in siblings, CLL risk alleles were found with high and similar prevalence in CLL patients and MBL siblings, suggesting that CLL risk loci predispose to clonal expansion of CLL phenotype cells in both low-count MBL and CLL. Vice versa, we did not find evidence for a particular association of CLL risk alleles with malignant progression from MBL to clinical CLL. Autonomous BCR signaling operates in both MBL and CLL as predicted by BCR stereotype, albeit at lesser strength in MBL. Acquired CLL-genetic changes, both copy number and sequence variants, are frequently subclonal in MBL but not in CLL. These findings support a stepwise CLL pathogenetic model wherein autonomous BCR signaling causes a non-malignant expansion of clonal CD5+ B cells in genetically predisposed individuals, followed by malignant progression to CLL after gradual acquisition of pathogenic genetic variants.

Structural Elements of the BCR Determine Permissiveness to Oncogenic RAS-Driven B-Cell Transformation

Background: RAS-pathway activating mutations (e.g. NRAS, KRAS, BRAF) invariably promote constitutive ERK-activation and are oncogenic drivers in B-cell acute lymphoblastic leukemia (B-ALL; Irving et al., 2014). Conversely, persistent ERK-activation can induce plasma cell differentiation (Yasuda et al., 2011), negative B-cell selection and cell death (Limnander et al., 2011) during B-cell development. In RAS-associated autoimmune leukoproliferative disorder (RALD), RAS-activating germline mutations cause lymphoproliferative disorder but not leukemia. The molecular determinants driving these divergent outcomes of RAS-activation have yet to be characterized. For these reasons, we here examined the mechanisms that enable permissiveness to RAS-driven transformation of B-cells. Results: Studying RAS-pathway activating mutations, we found that these genetic lesions are very common in B-cell malignancies derived from the earliest (e.g. B-ALL) and the latest (e.g. multiple myeloma) stages of B-cell development. In contrast, RAS-pathway lesions are exceedingly rare in B-cell receptor (BCR)-expressing B-cell malignancies, including mantle cell lymphoma, chronic lymphocytic leukemia, follicular lymphoma, Burkitt's lymphoma and diffuse large B-cell lymphoma (Figure 1). We also studied the consequence of oncogenic RAS-activation in mature B-cells, which express BCR. Consistent with previous findings that ERK-activation results in negative B-cell selection and cell death of B-cells (Limnander al., 2011), we found that expression of NRASG12D in mature B-cells resulted in upregulation of Prdm1, a tumor suppressor and mediator of negative B-cell selection and cell death (Hug et al., 2014; Setz et al., 2018). Altogether, our results suggest that mature B-cells are not permissive to RAS-driven transformation and prompted us to investigate whether expression of BCR counteracts constitutive ERK-activation resulting from oncogenic RAS signaling in B-cells. Reflecting incompatibility between oncogenic RAS-ERK activation and BCR-expressing malignant B-cells, expression of NRASG12D in BCR-positive mantle cell lymphoma cells transiently decreased surface expression of BCR but eventually led to depletion of cells from culture in growth competition assays. The non-receptor protein tyrosine kinase SYK is a key mediator of BCR signaling. Notably, treatment with small molecule inhibitor of SYK (entospletinib) largely reversed cell death caused by oncogenic RAS-signaling. Taken together, our results suggest that expression of an active BCR prevents permissiveness to constitutive ERK-activation. BCR consists of µ heavy chain (µHC) and conventional light chains (LCs; either kappa LC or lambda LC). Given that expression of a functional BCR opposes constitutive ERK-activation, this raised the question of whether structural elements of BCR are involved in determining outcome of RAS-activation. To this end, we performed CRISPR/Cas9-based gene editing to disrupt the constant region of immunoglobulin heavy chain (IGHM) or the constant region of the immunoglobulin kappa LC (IGKC) in kappa LC-expressing mantle cell lymphoma cells. While CRISPR-mediated deletion of IGHM reduced surface expression of IgM (μHC), this did not rescue cells from call death caused by oncogenic RAS-ERK activation. On the contrary, genetic ablation of IGKC enabled permissiveness to oncogenic RAS-signaling and prevented cell death resulting from constitutive ERK-activation. Hence, these findings showed that expression of kappa LC opposes constitutive RAS-ERK signaling in B-cell lymphoma cells. Conclusions: Surface expression of BCR is conserved in most mature B-cell lymphomas. Recent studies suggest that constitutive RAS-ERK signaling can override BCR requirement in B-cell lymphoma and that B-cell lymphoma cells acquiring RAS mutations may lose selective pressure to express a functional BCR (Varano et al., 2017). Here, our results demonstrated that conventional LCs of the BCR determine permissiveness to RAS-driven malignant transformation of B-cells. Conventional LCs were previously shown to inhibit the autonomous signaling capacity of BCR (Meixlsperger et al., 2007). We here showed that conventional LCs of BCR counteract oncogenic RAS signaling, providing a mechanistic basis of the low frequency of RAS-pathway activating mutations observed in BCR-expressing B-cell malignancies. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Subjective-Physiological Coherence during Food Consumption in Older Adults.

Subjective-physiological emotional coherence is thought to be associated with enhanced well-being, and a relationship between subjective-physiological emotional coherence and superior nutritional status has been suggested in older populations. However, no study has examined subjective-physiological emotional coherence among older adults while tasting food. Accordingly, the present study compared subjective-physiological emotional coherence during food consumption among older and younger adults. Participants consumed bite-sized gel-type foods with different flavors and provided their subjective ratings of the foods while their physiological responses (facial electromyography (EMG) of the corrugator supercilia, masseter, and suprahyoid, and other autonomic nervous system signals) were simultaneously measured. Our primary findings were that (1) the ratings of liking, wanting, and valence were negatively correlated with corrugator EMG activity in older and young adult participants; (2) the positive association between masseter EMG activity and ratings of wanting/valence was weaker in the older than in the young adult group; and (3) arousal ratings were negatively correlated with corrugator EMG activity in the older group only. These results demonstrate commonalities and differences in subjective-physiological emotional coherence during food intake between older and young adults.

An orderly sequence of autonomic and neural events at transient arousal changes

Resting-state functional magnetic resonance imaging (rsfMRI) allows the study of functional brain connectivity based on spatially structured variations in neuronal activity. Proper evaluation of connectivity requires removal of non-neural contributions to the fMRI signal, in particular hemodynamic changes associated with autonomic variability. Regression analysis based on autonomic indicator signals has been used for this purpose, but may be inadequate if neuronal and autonomic activities covary. To investigate this potential co-variation, we performed rsfMRI experiments while concurrently acquiring electroencephalography (EEG) and autonomic indicator signals, including heart rate, respiratory depth, and peripheral vascular tone. We identified a recurrent and systematic spatiotemporal pattern of fMRI (named as fMRI cascade), which features brief signal reductions in salience and default-mode networks and the thalamus, followed by a biphasic global change with a sensory-motor dominance. This fMRI cascade, which was mostly observed during eyes-closed condition, was accompanied by large EEG and autonomic changes indicative of arousal modulations. Importantly, the removal of the fMRI cascade dynamics from rsfMRI diminished its correlations with various signals. These results suggest that the rsfMRI correlations with various physiological and neural signals are not independent but arise, at least partly, from the fMRI cascades and associated neural and physiological changes at arousal modulations.

PMON255 Inflammation and Oxidative Stress in Seminal Plasma: Role of Free Light Chains of Immunoglobulins and Antioxidants as Biomarkers in Male Infertility

Abstract Oxidative and inflammatory damage are underlying mechanisms in various conditions related to male infertility, including varicocele. Free light chains of immunoglobulins (FLCs) are considered markers of low-grade inflammation in various conditions. Nowadays it is known that they may play enzymatic and proteolytic activities; they may activate complement cascade and mast cells, with crucial role in the development of contact sensibility; they may also inhibit the autonomous signaling ability of the B-cell receptor. Their detection in biological fluids may be useful as biomarker of inflammation or altered immune response. Coenzyme Q10 (CoQ10), a lipidic antioxidant and component of the mitochondrial respiratory chain, is involved in spermatozoa energy metabolism and motility. Therefore, we aimed to evaluate FLCs seminal levels in patients with varicocele in comparison to control subjects and to correlate them with CoQ10 and Total Antioxidant Capacity (TAC) in human semen. A total of 65 patients were enrolled, aged 18-50 years. Semen analysis was performed; patients were divided into three groups: controls (CTRL), 12 normozoospermic patients aged 34 (33-41) years; varicocele (VAR), 29 patients aged 33 (26-37) years; idiopathic (IDIO), 24 oligo-, astheno- and oligoasthenozoospermic patients aged 37 (33.5-40.5) years. FLCs were assayed by turbidimetric method; CoQ10 by HPLC; TAC by spectrophotometric method. Λ FLCs showed a trend towards higher levels in VAR vs CTRL and IDIO. VAR showed a trend towards lower κ FLCs levels vs the other two groups. Comparing κ/λ ratio VAR showed significantly lower levels vs CTRL and IDIO (Mean±SEM 3.18±0.75; 8.28±1.71; 8.90±2.94, respectively; p&amp;lt;0.05). CoQ10 seminal levels showed higher levels in VAR and IDIO compared to CTRL (57.7±4.13; 65.50±6.95 and 38.30±3.35 mcg/ml, respectively; p&amp;lt;0.05) No different TAC values were found between groups. Our data confirm lower levels of κ/λ ratio in VAR as previously reported and suggest a possible application as clinical biomarkers for male infertility. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Phosphorylation, compartmentalization, and cardiac function.

Protein phosphorylation is a fundamental element of cell signaling. First discovered as a biochemical switch in glycogen metabolism, we now know that this posttranslational modification permeates all aspects of cellular behavior. In humans, over 540 protein kinases attach phosphate to acceptor amino acids, whereas around 160 phosphoprotein phosphatases remove phosphate to terminate signaling. Aberrant phosphorylation underlies disease, and kinase inhibitor drugs are increasingly used clinically as targeted therapies. Specificity in protein phosphorylation is achieved in part because kinases and phosphatases are spatially organized inside cells. A prototypic example is compartmentalization of the cyclic adenosine 3',5'-monophosphate (cAMP)-dependent protein kinase A through association with A-kinase anchoring proteins. This configuration creates autonomous signaling islands where the anchored kinase is constrained in proximity to activators, effectors, and selected substates. This article primarily focuses on A kinase anchoring protein (AKAP) signaling in the heart with an emphasis on anchoring proteins that spatiotemporally coordinate excitation-contraction coupling and hypertrophic responses.

Underwater Navigation of AUVs Using Uncorrelated Measurement Error Model of USBL

&lt;i&gt;This article presents a modeling method for the uncorrelated measurement error of the ultra-short baseline (USBL) acoustic positioning system for aiding navigation of underwater vehicles. The Mahalanobis distance (MD) and principal component analysis are applied to decorrelate the errors of USBL measurements, which are correlated in the x- and y-directions and vary according to the relative direction and distance between a reference station and the underwater vehicles. The proposed method can decouple the radial-direction error and angular direction error from each USBL measurement, where the former and latter are independent and dependent, respectively, of the distance between the reference station and the vehicle. With the decorrelation of the USBL errors along the trajectory of the vehicles in every time step, the proposed method can reduce the threshold of the outlier decision level. To demonstrate the effectiveness of the proposed method, simulation studies were performed with motion data obtained from a field experiment involving an autonomous underwater vehicle and USBL signals generated numerically by matching the specifications of a specific USBL with the data of a global positioning system. The simulations indicated that the navigation system is more robust in rejecting outliers of the USBL measurements than conventional ones. In addition, it was shown that the erroneous estimation of the navigation system after a long USBL blackout can converge to the true states using the MD of the USBL measurements. The navigation systems using the uncorrelated error model of the USBL, therefore, can effectively eliminate USBL outliers without loss of uncontaminated signals.&lt;/i&gt;

Prediction of Specific Anxiety Symptoms and Virtual Reality Sickness Using In Situ Autonomic Physiological Signals During Virtual Reality Treatment in Patients With Social Anxiety Disorder: Mixed Methods Study.

BackgroundSocial anxiety disorder (SAD) is the fear of social situations where a person anticipates being evaluated negatively. Changes in autonomic response patterns are related to the expression of anxiety symptoms. Virtual reality (VR) sickness can inhibit VR experiences.ObjectiveThis study aimed to predict the severity of specific anxiety symptoms and VR sickness in patients with SAD, using machine learning based on in situ autonomic physiological signals (heart rate and galvanic skin response) during VR treatment sessions.MethodsThis study included 32 participants with SAD taking part in 6 VR sessions. During each VR session, the heart rate and galvanic skin response of all participants were measured in real time. We assessed specific anxiety symptoms using the Internalized Shame Scale (ISS) and the Post-Event Rumination Scale (PERS), and VR sickness using the Simulator Sickness Questionnaire (SSQ) during 4 VR sessions (#1, #2, #4, and #6). Logistic regression, random forest, and naïve Bayes classification classified and predicted the severity groups in the ISS, PERS, and SSQ subdomains based on in situ autonomic physiological signal data.ResultsThe severity of SAD was predicted with 3 machine learning models. According to the F1 score, the highest prediction performance among each domain for severity was determined. The F1 score of the ISS mistake anxiety subdomain was 0.8421 using the logistic regression model, that of the PERS positive subdomain was 0.7619 using the naïve Bayes classifier, and that of total VR sickness was 0.7059 using the random forest model.ConclusionsThis study could predict specific anxiety symptoms and VR sickness during VR intervention by autonomic physiological signals alone in real time. Machine learning models can predict the severe and nonsevere psychological states of individuals based on in situ physiological signal data during VR interventions for real-time interactive services. These models can support the diagnosis of specific anxiety symptoms and VR sickness with minimal participant bias.Trial RegistrationClinical Research Information Service KCT0003854; https://cris.nih.go.kr/cris/search/detailSearch.do/13508

Pathogenic variants of sphingomyelin synthase SMS2 disrupt lipid landscapes in the secretory pathway.

Sphingomyelin is a dominant sphingolipid in mammalian cells. Its production in the trans-Golgi traps cholesterol synthesized in the ER to promote formation of a sphingomyelin/sterol gradient along the secretory pathway. This gradient marks a fundamental transition in physical membrane properties that help specify organelle identify and function. We previously identified mutations in sphingomyelin synthase SMS2 that cause osteoporosis and skeletal dysplasia. Here, we show that SMS2 variants linked to the most severe bone phenotypes retain full enzymatic activity but fail to leave the ER owing to a defective autonomous ER export signal. Cells harboring pathogenic SMS2 variants accumulate sphingomyelin in the ER and display a disrupted transbilayer sphingomyelin asymmetry. These aberrant sphingomyelin distributions also occur in patient-derived fibroblasts and are accompanied by imbalances in cholesterol organization, glycerophospholipid profiles, and lipid order in the secretory pathway. We postulate that pathogenic SMS2 variants undermine the capacity of osteogenic cells to uphold nonrandom lipid distributions that are critical for their bone forming activity.

Abstract 053: Increased Arterial Pressure Precedes Renal Dysfunction And Elevated Renal Sympathetic And Afferent Nerve Activity In Deoxycorticosterone-Salt Rat

Hypertension (HTN) is a primary risk factor for cardiovascular disease - the leading cause of death worldwide. HTN is multifaceted but a key factor is aberrant autonomic signaling, including renal sympathetic (RSNA) and afferent nerve activity (ARNA). ARNA is elevated in several models of HTN and renal afferent nerve ablation can mitigate the HTN. It is unknown if ARNA rises before changes in blood pressure (BP) or renal function in HTN. This study aimed to determine when renal nerve activity changes vis a vis BP and renal function in a model of salt sensitive HTN. We hypothesized increased ARNA and RSNA would coincide with a rise in BP and decline in renal function. To test this hypothesis, 24 uninephrectomized male Sprague Dawley rats (300g) received a deoxycorticosterone acetate (DOCA; 100mg, SC) implant and 0.9% NaCl drinking water. Renal function (serum creatinine; Cr) and systolic BP (SBP; tail cuff) were measured weekly. Animals were randomly allocated to acute renal nerve recording on protocol day (D) 0, 7, 14, or 21. Multiunit RSNA and ARNA were recorded serially, as ARNA was isolated after sectioning proximal side. RSNA is expressed as bursts/heartbeat, ARNA and expressed as %ARNA max (response to intrapelvic capsaicin). Data were analyzed by one-way ANOVA with Bonferroni post hoc. Data presented as mean±SEM; *p&lt;.05 vs D0. SBP (mmHg) progressively increased on D7 (*152 ± 3), D14 (*171 ± 3), and D21 (*197 ± 7) vs D0 (138 ± 3). ARNA (%) increased at D14 (*23.0 ± 4.7) and D21 (*34.2 ± 4.4) vs D0 (10.1 ± 0.8), but no changes were detected at D7 (13.4 ± 2.1). No changes in RSNA from D0 were detected. Cr (mg/dL) increased at D14 (*1.2 ± 0.1) and D21 (*1.2 ± 0.1) vs D0 (0.9 ± 0.0), indicating a decline in kidney function. These data do not support the hypothesis that ARNA and RSNA increase in parallel to increasing BP. Rather, BP appears to initially increase independent of detectable changes in RSNA, ARNA or renal dysfunction. Interestingly, renal dysfunction and elevated ARNA were detected on D14, which coincides with our previous report that renal inflammation is first detected at D14. These data do not support a causal role of ARNA in this model. Rather, ARNA may subsequently drive disease progression. Further studies on the cause of increased ARNA and inflammation interaction are underway.

BDNF-dependent modulation of axonal transport is selectively impaired in ALS

Axonal transport ensures long-range delivery of essential cargoes between proximal and distal compartments, and is needed for neuronal development, function, and survival. Deficits in axonal transport have been detected at pre-symptomatic stages in the SOD1G93A and TDP-43M337V mouse models of amyotrophic lateral sclerosis (ALS), suggesting that impairments in this critical process are fundamental for disease pathogenesis. Strikingly, in ALS, fast motor neurons (FMNs) degenerate first whereas slow motor neurons (SMNs) are more resistant, and this is a currently unexplained phenomenon. The main aim of this investigation was to determine the effects of brain-derived neurotrophic factor (BDNF) on in vivo axonal transport in different α-motor neuron (MN) subtypes in wild-type (WT) and SOD1G93A mice. We report that despite displaying similar basal transport speeds, stimulation of wild-type MNs with BDNF enhances in vivo trafficking of signalling endosomes specifically in FMNs. This BDNF-mediated enhancement of transport was also observed in primary ventral horn neuronal cultures. However, FMNs display selective impairment of axonal transport in vivo in symptomatic SOD1G93A mice, and are refractory to BDNF stimulation, a phenotype that was also observed in primary embryonic SOD1G93A neurons. Furthermore, symptomatic SOD1G93A mice display upregulation of the classical non-pro-survival truncated TrkB and p75NTR receptors in muscles, sciatic nerves, and Schwann cells. Altogether, these data indicate that cell- and non-cell autonomous BDNF signalling is impaired in SOD1G93A MNs, thus identifying a new key deficit in ALS.

Mutant KRAS modulates colorectal cancer cells invasive response to fibroblast-secreted factors through the HGF/C-MET axis.

Genetic alterations influencethe malignant potential of cancer cells, and so does the tumor microenvironment. Herein, we combined the study of KRAS oncogenic effects in colorectal cancer cells with the influence of fibroblast-derived factors. Results revealed that mutant KRAS regulates cell fate through both autonomous and nonautonomous signaling mechanisms. Specifically, processes such as proliferation and cell-cell aggregation were autonomously controlled by mutant KRAS independently of the stimulation with fibroblasts conditioned media. However, cancer cell invasion revealed to be a KRAS-dependent nonautonomous effect, resulting from the cooperation between fibroblast-derived HGF and mutant KRAS regulation of C-MET expression. C-MET downregulation upon KRAS silencing rendered cells less responsive to HGF and thus less invasive. Yet, in one cell line, KRAS inhibition triggered invasion upon stimulation with fibroblasts conditioned media. Inhibition of PIK3CA oncogene did not promote invasion, thus showing a KRAS-specific effect. Moreover, the invasive capacity also depended on the HGF-C-MET axis. Overall, our study awards oncogenic KRAS an important role in modulating the response to fibroblast-secreted factors either by promoting or impairing invasion, and depicts the HGF-C-MET axis as a putative therapeutic target to impair the invasive properties of mutant KRAS cancer cells.

A perinuclear calcium compartment regulates cardiac myocyte hypertrophy

The pleiotropic Ca2+/calmodulin-dependent phosphatase calcineurin is a key regulator of pathological cardiac myocyte hypertrophy. The selective activation of hypertrophic calcineurin signaling under stress conditions has been attributed to compartmentation of Ca2+ signaling in cardiac myocytes. Here, perinuclear signalosomes organized by the scaffold protein muscle A-Kinase Anchoring Protein β (mAKAPβ/AKAP6β) are shown to orchestrate local Ca2+ transients, inducing calcineurin-dependent NFATc nuclear localization and myocyte hypertrophy in response to β-adrenergic receptor activation. Fluorescent biosensors for Ca2+ and calcineurin and protein kinase A (PKA) activity, both diffusely expressed and localized by nesprin-1α to the nuclear envelope, are used to define an autonomous mAKAPβ signaling compartment in adult and neonatal rat ventricular myocytes. Notably, β-adrenergic-stimulated perinuclear Ca2+ and PKA and CaN activity transients depended upon mAKAPβ expression, while Ca2+ elevation and PKA and CaN activity in the cytosol were mAKAPβ independent. Buffering perinuclear cAMP and Ca2+ prevented calcineurin-dependent NFATc nuclear translocation and myocyte hypertrophy, without affecting cardiac myocyte contractility. Additional findings suggest that the perinuclear Ca2+ transients were mediated by signalosome-associated ryanodine receptors regulated by local PKA phosphorylation. These results demonstrate the existence of a functionally independent Ca2+ signaling compartment in the cardiac myocyte regulating hypertrophy and provide a premise for targeting mAKAPβ signalosomes to prevent selectively cardiac hypertrophy in disease.

Equivalent pupillary mimicry in younger and older adults.

Pupillary contagion is a form of autonomic mimicry in which faces with dilated pupils elicit larger pupils in observers whereas faces with constricted pupils elicit smaller pupils. Autonomic reactivity may be fundamental to higher order social processes, yet older adults may be less likely to register other's autonomic signals. We explored pupillary contagion in younger and older adult observers. We presented younger and older observers with partial-face photographs of women with the pupils manipulated to be small, medium, or large. The faces were either young (20s) or old (70s). There were two tasks: To judge the model's age and to judge which pupil was larger. In the pupil judgment task, the magnitude of response was lower in older adults than in younger adults, but both younger and older observers showed equivalent pupillary contagion. In the age judgment task, which did not draw attention to the pupils, we found no evidence of pupillary contagion in either age-group. Registration of the autonomic signal of pupil dilation does not appear to be impaired in older adults. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Tip60 acetylation of histone H3K4 temporally controls chromosome passenger complex localization.

The Chromosome Passenger Complex (CPC) generates chromosome autonomous signals that regulate mitotic events critical for genome stability. Tip60 is a lysine acetyltransferase that is a tumor suppressor and is targeted for proteasomal degradation by oncogenic papilloma viruses. Mitotic regulation requires the localization of the CPC to inner centromeres, which is driven by the Haspin kinase phosphorylating histone H3 on threonine 3 (H3T3ph). Here we describe how Tip60 acetylates histone H3 at lysine 4 (H3K4ac) to block both the H3T3ph writer and the reader to ensure that this mitotic signaling cannot begin before prophase. Specifically, H3K4ac inhibits Haspin phosphorylation of H3T3 and prevents binding of the Survivin subunit to H3T3ph. Tip60 acetylates H3K4 during S/G2 at centromeres. Inhibition of Tip60 allows the CPC to bind centromeres in G2 cells, and targeting of Tip60 to centromeres prevents CPC localization in mitosis. The H3K4ac mark is removed in prophase by HDAC3 to initiate the CPC localization cascade. Together, our results suggest that Tip60 and HDAC3 temporally control H3K4 acetylation to precisely time the targeting of the CPC to inner centromeres.

An Unexplored Role for MMP-7 (Matrix Metalloproteinase-7) in Promoting Gut Permeability After Ischemic Stroke.

Poststroke infections are common complications of stroke and are highly associated with poor outcomes for patients. Stroke induces profound immunodepression coupled with alterations to autonomic signaling, which together render the body more susceptible to infection from without (nosocomial/community-acquired infection) and from within (commensal bacterial infection). Critical to the hypothesis of commensal infection is the phenomenon of poststroke gut permeability and gut dysbiosis. Few studies have provided adequate explanations for the mechanisms underlying the molecular alterations that produce a more permeable gut and perturbed gut microbiota after stroke. A dysregulation in the production of matrix MMP-7 (metalloproteinase-7) may play a critical role in the progression of gut permeability after stroke. By cleaving junctional and extracellular matrix proteins, MMP-7 is capable of compromising gut barrier integrity. Because of MMP-7's unique abundance in the small intestine and its capacity to be induced in states of bacterial invasion and inflammation, along with its unique degradative capability, MMP-7 may be crucially important to the progression of gut permeability after ischemic stroke.

Laser Acupuncture and Dynamic Pulse Reaction Quantified by Smartphone:Two Experimental Examples.

The dynamic pulse reaction (Nogier reflex), Reflex-Auriculo-Cardiac (RAC), or vascular autonomic signal is a physiologic phenomenon that is not fully accepted and widely understood in contemporary medical practice. In order to provide appropriate scientific evidence for better understanding, qualitative and-above all-quantitative research in this area is necessary. In this short report, 2 recordings of dynamic pulse reactions based on an analysis performed on a modified smartphone are demonstrated during laser acupuncture experiments using stimulation at ear acupuncture points.

Involvement of the liver-gut peripheral neural axis in nonalcoholic fatty liver disease pathologies via hepatic HTR2A.

ABSTRACTSerotonin (5-HT) is one of the key bioamines of nonalcoholic fatty liver disease (NAFLD). Its mechanism of action in autonomic neural signal pathways remains unexplained; hence, we evaluated the involvement of 5-HT and related signaling pathways via autonomic nerves in NAFLD. Diet-induced NAFLD animal models were developed using wild-type and melanocortin 4 receptor (MC4R) knockout (MC4RKO) mice, and the effects of the autonomic neural axis on NAFLD physiology, 5-HT and its receptors (HTRs), and lipid metabolism-related genes were assessed by applying hepatic nerve blockade. Hepatic neural blockade retarded the progression of NAFLD by reducing 5-HT in the small intestine, hepatic HTR2A and hepatic lipogenic gene expression, and treatment with an HTR2A antagonist reproduced these effects. The effects were milder in MC4RKO mice, and brain 5-HT and HTR2C expression did not correlate with peripheral neural blockade. Our study demonstrates that the autonomic liver-gut neural axis is involved in the etiology of diet-induced NAFLD and that 5-HT and HTR2A are key factors, implying that the modulation of the axis and use of HTR2A antagonists are potentially novel therapeutic strategies for NAFLD treatment. This article has an associated First Person interview with the first author of the paper.