The aim of this study was to investigate the role of a synthetic dermal matrix, Biodegradable Temporising Matrix (BTM), for coverage of complex wounds. The authors defined complex wounds as wounds not amenable to reconstruction with skin grafting alone due to an inherent avascularity of the wound bed, such as the presence of exposed bone, tendinous or neural structures.A retrospective review of a prospectively maintained database of complex wounds as defined above was carried out. Clinical and operative notes were reviewed along with review of an extensive photographic database demonstrating wound healing progress using staged dermal matrix (BTM) and autologous skin graft reconstruction.55 patients were identified who underwent staged dermal matrix and autologous skin graft reconstruction for complex wounds affecting a wide variety of patient demographics, treatment indications and body sites. Wound aetiology varied between burn injury, non-burn related trauma including degloving injury and infective complications. We discuss caveats relating to successful application of a dermal matrix, technique tips, prevention and management of complications.Dermal substitutes play an integral role in providing biological wound cover for avascular wound beds which may otherwise require complex distant flap or microsurgical free flap reconstruction. BTM is a completely synthetic dermal matrix comprised of a 2-mm-thick sheet of biodegradable polyurethane foam bonded to a non-biodegradable polyurethane sealing membrane. Our department has developed significant expertise in the use of BTM throughout its development from initial animal studies through to recent human clinical trials. The synthetic composition of BTM does not require rapid neo-vascularisation for its integrity or survival. As such, two-stage BTM reconstruction has proven robustness in the face of unfavourable wounds compared with other popular dermal matrices, physiologically covering avascular structures, allowing for early graft take, expediting rehabilitation and mobilisation with excellent scar cosmesis and limited contracture formation.Dermal matrices such as BTM play an important role in complex wound healing, frequently achieving excellent results with a low complication profile. BTM has been used successfully in cases where biological matrices would not routinely be considered as demonstrated by this clinical series. It has provided a valuable alternative to free-tissue transfer in patients with significant co-morbidities, vascular insufficiency and/or those for whom long operations are undesirable.


The COVID-19 pandemic in NYC, the epicenter of the US crisis, revealed indisputable evidence that social determinants of health  and disparities in comorbid health risk factors produce higher burdens of disease and death among racial and ethnic populations. We conducted a needs assessment of SDoH among 1400 patients in several ambulatory care clinics to explore the impact among older adults, across different clinical populations. Among older adults with HIV (OAH), we found lower rates of food and housing insecurity compared to older adults without HIV. Despite higher levels of COVID knowledge and prevention adherence, we also found significantly higher levels of isolation, loneliness, depressive symptoms, and anxiety among OAHs compared to those without HIV. Access to Ryan White entitlements did buffer some impacts but preexisting high burdens of mental health issues were exacerbated, perhaps due to heightened perceptions of increased vulnerability to COVID-19.


There is an abundance of evidence for the widespread presence of this spinal reflex arch originating from virtually every visceral organ and thus having a substantial role in blood pressure control. Additionally, this review highlights specific endogenous eicosanoid species, which modulate the activity of afferent fibers involved in this reflex, through their interactions with transient receptor potential (TRP) cation channels.The spinal cord is an important integrative center for blood pressure control. Spinal sensory fibers send projections to sympathetic preganglionic neurons of the thoracic spinal cord and drive sympathetically-mediated increases in blood pressure. While these reflexes responses occur in able-bodied individuals, they are exaggerated following interruption of descending control – such as occurs following spinal cord injury. Similar reflex control of blood pressure may exist in disease states, other than spinal cord injury, where there is altered input to sympathetic preganglionic neurons. This review primarily focuses on mechanisms wherein visceral afferent information traveling  Physiological mechanisms of blood pressure control involve complex interactions between neural and humoral factors, which work hand-in-hand to ensure optimal delivery of nutrient rich blood to all the cells of the body. In regulating blood pressure, particularly important are the effects of the sympathetic nervous system. Sympathetic postganglionic neurons innervate blood vessels to regulate vascular smooth muscle tone and peripheral resistance. The rostral ventrolateral medulla is arguably one of the most critical cites for establishing baseline blood pressure as it sends tonic impulses to sympathetic preganglionic neurons located within the intermediolateral cell column of the thoracic spinal cord. Although the spinal cord is often described as a mere relay station between the brainstem and the periphery, clinical and experimental evidence highlights the important role of intraspinal reflex circuits in blood pressure regulation.via spinal nerves influences sympathetic nerve activity and blood pressure. Special emphasis is placed on descending inhibitory control of sympathetic preganglionic neurons and how interruption of this inhibition, as occurs following spinal cord injury, leads to aberrant spinal reflex control of blood pressure. Additionally, we propose that the spinal sensory input may contribute to tonic control of blood pressure in able bodied individuals. Finally, we highlight endogenous modulators of this neurocircuitry and their potential roles in regulation of blood pressure in healthy and diseased states.In this review we summarize available evidence that suggests intraspinal reflex circuits can directly and independently orchestrate profound reflex-mediated changes in blood pressure, especially when descending inhibitory pathways are interrupted, as in patients with spinal cord injuries. We primarily focus on mechanisms wherein visceral afferent information traveling via a spinal reflex arc – independent of central processing within the brain. The effect of visceral stimulation on systemic blood pressure was first empirically demonstrated in seminal works by Sir Charles Sherrington. He showed that distension of hollow visceral organs such as the urethra, bile duct or rectum in spinalized or decerebrate animals led to moderate increases in blood pressure, which were associated with a series of involuntary motor responses  and spinal pathways. Stimulation of cranial afferent nerve fibers is generally associated with cardiovascular depressor responses, while spinal afferent stimulation generally evokes pressor responses  within and surrounding visceral organs, are relayed to the central nervous system esponses . First oesponses . Within nal cord  , the lower visceral organs are arguably less innervated by vagal afferent fibers and are densely innervated by thoracolumbar and/or sacral (pelvic) afferent fibers, whose cells bodies are located within dorsal root ganglia. The nerve bundles of these spinal sensory neurons are composed of thinly myelinated Aδ and unmyelinated C fibers ,b. TheseVisceral mechano sensation has been extensively studied and mechanosensitive afferent fibers arising from the viscera are important drivers of sympathetic reflexes .Low threshold mechanosensitive afferent fibers innervate mucosal membranes and muscle layers of the visceral organs and are tonically active . They arHigh threshold mechanosensitive afferent fibers innervate blood vessels in the mesentery and mucosal membranes and muscle layers of visceral organs. These fibers are involved in detecting urgency, changes in mesenteric arterial pressure and mechanically induced pain.Silent mechanosensitive afferent fibers are mostly involved in signaling pathological visceral changes . These fIt is well established that mechanosensitive Aδ and C-fibers drive sympathetic responses to mechanical stimulation of the viscera . HoweverThe above described functional organization for processing of afferent information exists within most spinal segments and highlights potential of intraspinal circuitry to influence spinal visceral reflexes originating from various spinal segments.As described above, the viscero-sympathetic reflex, similar to spinal motor reflexes, is integrated at the level of the spinal cord. Various visceral reflex arcs are contained within the spinal cord and are involved in normal gastrointestinal and urogenital physiology. As indicated below, activation of most spinal visceral reflexes is preserved following upper thoracic spinal cord injury and is associated with increases in systemic blood pressure.Micturition reflex: During the storage phase of the micturition reflex, pelvic sensory afferents send slow impulses into the spinal cord, which drive involuntary contraction of the internal urethral sphincter and relaxation of the detrusor muscle, both of which are needed to facilitate urine retention and are mediated via sympathetic efferent fibers. Importantly, bladder filling is associated with a progressive increase in activity of other regional sympathetic efferent fibers and in systemic blood pressure  results in reflex increases in sympathetic nerve activity and blood pressure . Sympathetic fibers innervating the bladder neck, prostate, seminal vesicles, and vas deferens are activated and drive the emission phase of ejaculation  across the plasma membrane. TRP channels are activated by a wide range of physiological and pathophysiological signals and respond to both chemical ligands and physical stimuli. In addition, many of the mammalian TRP channels are thermo-sensitive and can be directly activated by cold or hot temperatures  family. To date, 28 mammalian TRP channels have been identified  salt-sensitive hypertension  or can be indirectly activated  via three major enzyme systems: the cyclooxygenase (COX), lipoxygenase (LOX) and cytochrome P450 (CYP) pathways. Eicosanoid molecules play a critical role in inflammatory signaling (2 (PGE2), clearly indicate that binding of PGE2 to its G protein coupled receptors (EP1-4) on the surface of TRP channel expressing neurons promotes hyperalgesia by driving phosphorylation of TRP channels thereby increasing excitability to mechanical, thermal and chemical stimuli (reviewed in via direct interactions (Eicosanoids are bioactive metabolites derived from polyunsaturated fatty acids (PUFAs) and generated ignaling , and theractions . Such inractions .Interestingly, many of the eicosanoids that promote afferent hyperexcitability through interactions with TRP channels are derived from ω-6 PUFAs, which are abundant in Western diets. This has led us and others to hypothesize that dietary PUFA content may directly influence the excitability of TPR expressing C and Aδ fibers. Evidence to support this hypothesis was recently published by the laboratory of K.M. Hargeaves . Specifivia intraspinal circuitry. However, it is important to highlight that similar sympathetic responses can be generated by spinal afferent information arising from somatic afferents   . As in t reflex) . While i reflex) . However reflex) . This lein vivo electrophysiology recording will be an important tool for understanding the role of eicosanoids in altering spinal afferent excitability. Furthermore, additional studies into the role of dietary PUFA content and eicosanoid-TRP interactions in regulating baseline sympathetic nerve activity are needed.We have briefly highlighted the potential role of eicosanoid-TRP interactions in modulating the afferent limb of the viscero-sympathetic reflex circuitry. However, many mechanistic issues remain to be resolved and via chronic activation of the viscero-sympathetic reflex circuitry. Consistent with this notion, a large Danish cohort study found the risk for developing hypertension was significantly reduced among patients undergoing surgical colectomy when compared to patients undergoing other surgical procedures (It is notable that inflammatory conditions involving the viscera, such as inflammatory bowel disease, lead to the development of visceral afferent hyperexcitability . Interesocedures . Future ZM and CR: idea, writing, and editing. DO’L: idea and editing. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Kefir is an easy to administer per feeding tube probiotic yogurt that does not contain the risk of powdered probiotics, which may contaminate patient wounds or intravenous lines. Previous studies show patients taking probiotics may decrease hospital-acquired infections (HAI) although kefir has not been well studied. We hypothesized that kefir would be well tolerated and prevent infections among critically injured patients including patients with burn injury on enteral nutrition (EN).We performed a retrospective review of adult critically injured patients at a level 1 trauma and burn center from January 2018 to March 2021 who received EN. Patients with a history of clostridium difficile (C. diff) were excluded. Patients who received kefir were given 120ml twice daily. The kefir protocol was improved with input from clinical stakeholders. The rate of C. diff, catheter-associated urinary tract infection (CAUTI), and central line-associated blood stream infection (CLABSI) were compared between patients who received kefir and those who did not. Incidence rate ratios (IRR) and corresponding 95% confidence intervals were calculated to assess differences in these rates.3,814 patients met criteria, 545 of whom received kefir (14%). Suggested improvements to the kefir protocol by stakeholders were changing flavored to plain kefir to decrease the amount of carbohydrate, change to lactose-free kefir to improve usage in lactose intolerant patients, and educate nurses on flushing feeding tubes to avoid clogs.None of the incidence rates of HAI were significantly different between patients who received kefir and those who did not (Table 1). Crude IRRs suggest that C. diff infections may have occurred less frequently among patients who received kefir while the reverse occurred for CLABSI infections, though these results are not significant.The kefir implementation was refined by stakeholder feedback. Although no clear benefit of kefir was observed with HAI reduction, future research should investigate the potential association between kefir use and C. diff.


Background: Numerous studies have shown that trauma-focused psychological treatments for PTSD are effective .Results: Were preceded by changes in for most measures, changes in cognitive factors preceded changes in PTSD symptoms, but not vice versa.Study 2 Methods: Patient and therapist ratings of working alliance were taken repeatedly. Autoregressive, cross-lagged models were used to examine whether working alliance predicted PTSD symptom severity at the next assessment point and vice versa.Results: Higher patient and therapist working alliance was associated with lower PTSD symptoms. Crossed-lagged associations were found for therapist-rated alliance, but not for patient-rated alliance. A better working alliance at the start of treatment predicted treatment outcome. For most measures, changes in cognitive factors preceded changes in PTSD symptoms, but not vice versa.Conclusions: Study 1 extended previous findings that that cognitive and behavioural processes suggested by theoretical models of PTSD play a key role in driving symptom improvement during trauma-focused CBT. Changes in cognitive factors preceded symptom change. This result suggests that monitoring these processes during therapy may help therapist in maximising treatment outcomes in individual patients, and may also point to further ways of optimising treatments in general. Study 2 suggested that working alliance may be an important factor in setting the necessary conditions for effective treatment, but did not find evidence of a temporal precedence of changes in working alliance driving symptom change.


The ultimate goal for vaccination is the generation of a safe and effective immune response that protects against diseases. Adjuvants are included to enhance and direct vaccine responses. Precisely how adjuvants and indeed vaccine formulations influence the innate immune response and the subsequent cell- and antibody-mediated arms of the adaptive immune system are being characterized and clarified. Identification of safe but potent vaccine adjuvants, particularly those directed at mucosal sites, which generate long-lasting immunological memory and are effective against intracellular and extracellular pathogens is a high priority for public health and the health of veterinary species. After they are identified, tremendous amounts of work must be done to fine-tune adjuvant production, stabilization, and purification before commercialization. We launched this special issue to highlight recent basic research in vaccine formulation for infectious diseases and assess the adjuvant mechanism of action.Alcaligenes spp. that inhabit Peyer’s patches in the gut [Alcaligenes lipid A may be a safe and effective Th17-type adjuvant.The following is a synopsis of the results reported in seven original articles. Systemic or mucosal adjuvants that promote T helper 17 (Th17)-type immunity are being investigated. Chaffey et al., 2021) assessed whether an intramuscular injection of adjuvants with ovalbumin in mice could induce an antigen-specific Th17-type immune response [021 asses the gut . Ex vivoMucosal vaccines need potent mucosal adjuvants to be effective. Razim et al. developed and characterized nanoadjuvant candidates (NACs) composed of silicon oil and cationic detergents and organic solvents for intranasal application . They idAssessing whether FDA-approved drugs can be repurposed as vaccine adjuvants is an important way to potentially fast-track adjuvant approval. Miltefosine (MTF) is an FDA-approved anti-leishmaniasis drug. Lu et al. assessed its adjuvant potential by immunizing mice via an intraperitoneal injection with hemagglutinin from influenza followed by intranasal challenge . The micAvian metapneumovirus (aMPV) is a virus that is the causative agent of turkey rhinotracheitis in turkey flocks and swollen head syndrome in chickens leading to significant morbidity and mortality throughout China. Bao et al. performed intramuscular injections of inactivated aMPV/B strain LN16 formulated with commercially available immune-stimulating complexes  and measured the resultant immune response . They deFour review articles in this issue highlight the production, purification, and formulation of interferon-based biopharmaceuticals  , the effTaken together, this special issue contains 11 relevant research and review articles focused on various aspects of the adjuvant mechanism of action and novel vaccine formulations that should be explored further for use in human and veterinary vaccines.


FGFR inhibitors evolved as therapeutic options in cholangiocarcinoma and urothelial malignancies. Given the implications of FGFR pathway in various physiological functions, FGFR inhibitors are known to cause unique toxicities. In this review, we summarized the physiology of FGF/FGFR signaling and briefly discussed the possible mechanisms that could lead to FGFR inhibitor resistance and side effects. In addition, we proposed treatment guidelines for the management of FGFR-inhibitor-associated toxicities.FGFR2 fusions or rearrangement, and erdafitinib in metastatic urothelial carcinoma with FGFR2 and FGFR3 genetic aberrations, lead to their accelerated approval by the United States (USA) FDA. Along with these agents, many phase II/III clinical trials are currently evaluating the use of derazantinib, infigratinib, and futibatinib either alone or in combination with immunotherapy. Despite the encouraging results seen with FGFR inhibitors, resistance mechanisms and side effect profile may limit their clinical utility. A better understanding of the unique FGFR-inhibitor-related toxicities would invariably help us in the prevention and effective management of FGFR-inhibitor-induced adverse events thereby enhancing their clinical benefit. Herein, we summarized the physiology of FGF/FGFR signaling and briefly discussed the possible mechanisms that could lead to FGFR inhibitor resistance and side effects. In addition, we proposed treatment guidelines for the management of FGFR-inhibitor-associated toxicities. This work would invariably help practicing oncologists to effectively manage the unique toxicities of FGFR inhibitors.Fibroblast Growth Factor receptor (FGFR) pathway aberrations have been implicated in approximately 7% of the malignancies. As our knowledge of FGFR aberrations in cancer continues to evolve, FGFR inhibitors emerged as potential targeted therapeutic agents. The promising results of pemigatinib and infigratinib in advanced unresectable cholangiocarcinoma harboring  FGFR gene aberrations have been implicated in human malignancies . In . In 8]. TP53 gene thereby activating the cancer-associated fibroblasts promoting the tumorigenesis in FGFR aberrations . Given this expanding role of FGFR inhibitors in cancer care, a thorough knowledge on their unique side effects will aid in preventing unnecessary dose reductions and interruptions. It is important that patients be educated about these FGFR-inhibitor-related unique side effects and possible preventive mechanisms. Along with effective management of drug-related toxicities, a special focus is to be made on expanding our knowledge on resistance to FGFR-inhibitors. As determined in preclinical models, combination regimens such as synergistic inhibition of FGFR inhibitors and mTOR or MAPK pathway inhibitors may further be evaluated to bypass the resistance mechanisms.FGFR inhibitors are a unique class of emerging targeted therapies that have shown promising results in tumors harboring


Myocardial infarction leads to pathological changes in the heart, including loss of cardiomyocytes and degradation of the cardiac extracellular matrix (ECM). These changes result in wall thinning, infarct enlargement, and development of scar tissue. Such remodeling ultimately leads to end-stage heart failure ,2. To adIn order to address these limitations, Silveira-Filho et al.  investigMany studies in the field of cardiac tissue engineering address remodeling timepoints immediately after ischemic wounding. However, this acute approach is not realistic because most of the remodeling in clinical ischemic heart disease occurs undetected over long periods of time, leading to extensive ventricular thinning, scarring, and progressive heart failure. The experimental time course followed in this study attempted to recapitulate extensive remodeling in the validation of PECUU-ECM patches by addressing ischemic wounds 8 weeks after myocardial infarction. This study found PECUU-ECM patches were efficacious in reversing the negative remodeling seen in advanced stages of ischemia, offering great potential for clinical translation. Additionally, the authors’ rationale for incorporating PECUU patches within the biohybrid patch was its slower degradation rate relative to that of poly(ester urethane)urea (PEUU). This slower degradation rate ultimately led to better remodeling outcomes in post-infarcted rats given a PECUU patch compared to those given a PEUU patch over their infarcted hearts. Despite the slower rate of degradation, PECUU patches favorably did not form encapsulations typically caused by nondegradable material. These findings point to the importance of polymer selection in the development of cardiac patches. Notably, there are still some challenges this study needs to address in the future. The biohybrid patch used is mechanically compatible with rat myocardium, yet how these patches may influence the contractility and electrophysiology of human myocardium and whether the patches will withstand the pressures of a full adult human cardiac load upon implantation remain to be investigated . If implRecent advances in the field of cardiac tissue engineering include the use of human induced pluripotent stem cell (iPSC)-derived cardiomyocytes to produce engineered heart tissue (EHT) . In the The PECUU-ECM technology engineered by Silveira-Filho et al.  shows prThe authors have reported that they have no relationships relevant to the contents of this paper to disclose.


In mucosal barrier interfaces, flexible responses of gene expression to long-term environmental changes allow adaptation and fine-tuning for the balance of host defense and uncontrolled not-resolving inflammation. Epigenetic modifications of the chromatin confer plasticity to the genetic information and give insight into how tissues use the genetic information to adapt to environmental factors. The oral mucosa is particularly exposed to environmental stressors such as a variable microbiota. Likewise, persistent oral inflammation is the most important intrinsic risk factor for the oral inflammatory disease periodontitis and has strong potential to alter DNA-methylation patterns. The aim of the current study was to identify epigenetic changes of the oral masticatory mucosa in response to long-term inflammation that resulted in periodontitis.ROBO2, PTP4A3), cell adhesion (LPXN) and innate immune response . Enrichment analyses implied a role of epigenetic changes for vesicle trafficking gene sets.Genome-wide CpG methylation of both inflamed and clinically uninflamed solid gingival tissue biopsies of 60 periodontitis cases was analyzed using the Infinium MethylationEPIC BeadChip. We validated and performed cell-type deconvolution for infiltrated immune cells using the EpiDish algorithm. Effect sizes of DMPs in gingival epithelial and fibroblast cells were estimated and adjusted for confounding factors using our recently developed “intercept-method”. In the current EWAS, we identified various genes that showed significantly different methylation between periodontitis-inflamed and uninflamed oral mucosa in periodontitis patients. The strongest differences were observed for genes with roles in wound healing (Our results imply specific adaptations of the oral mucosa to a persistent inflammatory environment that involve wound repair, barrier integrity, and innate immune defense.The online version contains supplementary material available at 10.1186/s13148-021-01190-7. Genetic studies of chronic inflammatory diseases focused primarily on identifying DNA variants  that confer disease risk through genome-wide association studies . MorThe oral masticatory mucosa locates to the alveolus/tooth complex and the hard palate and comprises all parts of the oral mucosa that are involved in chewing. Its position at the entrance to the gastrointestinal tract and respiratory systems entails its direct exposition to a diverse microbiota and various other environmental stressors such as tobacco smoke. If the mucosal barrier is impaired, pathogens can invade into subjacent tissue layers and promote periodontal inflammation. Persisting inflammation causes gingival bleeding and leads to the loss of connective tissue and alveolar bone with subsequent tooth loss, which determines the clinical characteristics of the common oral inflammatory disease periodontitis. In the etiology of periodontitis, persistent gingival inflammation is the most important risk factor , followeCYP1B1 (cytochrome P450 family 1 subfamily B member 1) and AHRR (aryl-hydrocarbon receptor repressor). Several EWAS have identified these associations for cells of the alveolar  for the top 10 differentially methylated genes in the GO term “synaptic vesicle cycle”.


Traumatic brain injury (TBI) is common among older adults, with significant public health costs, and advanced age is a risk factor for poor outcomes after TBI. Older Veterans with TBI-related cognitive and emotional dysfunction without dementia may benefit from cognitive rehabilitation, particularly executive function training, and technology may promote optimal functioning for these patients by increasing access to such treatments. Dr. Kornblith will present pilot data on one such promising group intervention, Goal-Oriented Attentional Self-Regulation (GOALS), administered via in-home video telehealth. Themes gleaned from qualitative feedback collected throughout the intervention and post-treatment feedback questionnaires include the importance of communication and a smooth process with clear instructions for joining study sessions. Preliminary data suggest that only minor adaptions to the existing GOALS protocol are required for telehealth delivery and that delivering group-based executive function training to older TBI-exposed older Veterans with cognitive complaints via telehealth is feasible and acceptable.


Long-lasting changes in neural pain circuits precipitate the transition from acute to chronic pain in patients living with inflammatory bowel diseases (IBDs). While significant improvement in IBD therapy has been made to reduce inflammation, a large subset of patients continues to suffer throughout quiescent phases of the disease. Peripheral and central mechanisms contribute to the transition from acute to chronic pain during active disease and clinical remission. Lower mechanical threshold and hyperexcitability of visceral afferents induce gliosis in central pain circuits, leading to persistent visceral hypersensitivity (VHS). In the spinal cord, microglia, the immune sentinels of the central nervous system, undergo activation in multiple models of VHS. Using the Dextran Sodium Sulfate (DSS) model of colitis, we found that microglial G-CSF was able to sensitize colonic nociceptors that express the pain receptor TRPV1. While TRPV1+ nociceptors have been implicated in peripheral sensitization, their contribution to central sensitization via microglia remains unknown.Here we investigated the mechanisms of microglia activation to identify centrally acting analgesics for chronic IBD pain.Using Designer Receptors Exclusively Activated by Designer Drugs (DREADD) expressed in TRPV1-expressing visceral neurons that sense colonic inflammation, we tested whether neuronal activity was indispensable to control microglia activation and VHS. We then investigated the neuron-microglia signaling system involved in visceral pain chronification.We found that chemogenetic inhibition of TRPV1+ visceral afferents prevents microglial activation in the spinal cord and subsequent VHS in colitis mice. In contrast, chemogenetic activation, in the absence of colitis, enhanced microglial activation associated with VHS. We identified a purinergic signaling mechanism mediated by neuronal ATP and microglial P2RY12 receptor, triggering VHS in colitis. Inhibition of P2RY12 prevented microglial reactivity and chronic VHS post-colitis.Overall, these data provide novel insights into the central mechanisms of chronic visceral pain and suggest that targeting microglial P2RY12 signaling could be harnessed to relieve pain in IBD patients who are in remission.CCC


Retaining older adults in clinical trials has often been a challenge for researchers. Clinical trial procedures, aimed at improving fidelity, often offer barriers to frail older adults who have challenges traveling to medical centers and enduring long clinical assessment visits. During the COVID-19 pandemic, we modified the procedures of two randomized controlled trials. COMPASS: A novel transition program to reduce disability after stroke is a clinical trial examining the efficacy of a transition home program that provides home modifications and self-management strategies compared to stroke education. HARP: Removing home hazards for older adults living in affordable housing is a pragmatic trial examining the effectiveness of a home hazard removal program for residents of low-income housing. Modifications to the trials were designed to reduce human contact but in some cases reduced the burden on trial participants. Modified procedures addressed retention, assessment of endpoints and intervention methods.


Tumor heterogeneity is one of the hallmarks of glioblastoma multiforme (GBM). Morphology within a given GBM tumor can be extremely variable where some regions of the tumor have a soft, gel-like structure while other areas are dense and fibrous. Abnormal mechanical stress and tissue stiffening caused by cancer proliferation are believed to affect vascularity by compressing structurally weak blood vessels and restricting the supply of nutrients and oxygen to the tissue. These effects contribute to a hypoxic microenvironment that promotes disease progression and chemoresistance. The genetic and molecular mechanisms that govern tissue stiffness within GBM tumors, however, are largely unknown. Magnetic Resonance Elastography (MRE) is an emerging technique for quantifying tissue stiffness non-invasively.We have evaluated 10 GBM patients by MRE imaging obtained prior to surgical resection. During surgery, 2–7 stereotactically navigated biopsies were collected from locations within the tumor with varying degrees of measured stiffness. Biopsies were processed to extract RNA, proteins, polar metabolites and lipids. Biomolecules were analyzed on relevant -omics platforms .Differential expression and gene set enrichment analysis of patient paired biopsies indicate an overall increase in macrophage infiltration and extracellular matrix re-organization associated with increased tumor stiffness. Among the most highly upregulated genes in stiff tumor tissue were lymphatic endothelial hyaluronic acid receptor 1 (LYVE-1) and macrophage receptor with collagenous structure (MARCO), both of which have been associated with immune cell infiltration and tissue stiffness.Our preliminary findings offer novel insights into tumor morphology in GBM that can be inferred from imaging prior to surgery. This can be used to identify tumor regions with high risk of progression and infiltration, thereby informing and guiding surgical strategy and may ultimately lead to novel treatment strategies.


Roney et al. review recent advances in how axonal endolysosomal trafficking, distribution, and functionality maintain distal degradation capacity in neuronal health and become disrupted in several neurodegenerative diseases. Lysosomes serve as degradation hubs for the turnover of endocytic and autophagic cargos, which is essential for neuron function and survival. Deficits in lysosome function result in progressive neurodegeneration in most lysosomal storage disorders and contribute to the pathogenesis of aging-related neurodegenerative diseases. Given their size and highly polarized morphology, neurons face exceptional challenges in maintaining cellular homeostasis in regions far removed from the cell body where mature lysosomes are enriched. Neurons therefore require coordinated bidirectional intracellular transport to sustain efficient clearance capacity in distal axonal regions. Emerging lines of evidence have started to uncover mechanisms and signaling pathways regulating endolysosome transport and maturation to maintain axonal homeostasis, or “axonostasis,” that is relevant to a range of neurologic disorders. In this review, we discuss recent advances in how axonal endolysosomal trafficking, distribution, and lysosomal functionality support neuronal health and become disrupted in several neurodegenerative diseases. Lysosomes are dynamic, membrane-bound, acidic organelles that play a central role in the degradation of intracellular and extracellular cargos. More than 60 hydrolytic enzymes that are active under acidic environments within the lysosomal lumen facilitate degradation of complex biological macromolecules, including proteins, lipids, nucleic acids, and carbohydrates . LysosomNeurons are highly polarized cells consisting of a cell body, complex dendritic arbors, and a single long axon with extensive branches and terminals that can span several tens to hundreds of centimeters in length. Given this extended morphology, neurons require efficient bidirectional transport mechanisms to coordinate clearance of endocytic and autophagic cargos generated distally by lysosomes that are relatively enriched in the cell body. Efficient transport is critically important in maintaining neuron growth, survival, and function but particularly challenging in distal axons and terminal branching regions. Accumulation of autophagic vacuoles (AVs) and lysosome-like organelles characterizes axonal pathology in several neurodegenerative diseases associated with lysosome dysfunction, reflecting disruptions at various steps in the maturation and trafficking of endolysosomal and autophagic organelles . In thisLysosomes serve as terminal degradation hubs for endocytic and autophagic components. Extracellular materials internalized by endocytosis, intracellular components sequestered by autophagy, and newly synthesized lysosomal proteins reach endolysosomes through highly regulated trafficking routes. Endolysosomal trafficking from early endosomes (EEs) to late endosomes (LEs) and finally to mature lysosomes is essential for delivering target endosomal proteins for degradation . TransitThe unique compartmental features of neurons add yet another layer of complexity to lysosome maturation. As in other cell types, neuronal lysosomes receive cargos through the endocytic, autophagic, and biosynthetic pathways. Because of their extreme morphological features, neurons require coordinated bidirectional transport mechanisms to maintain a steady-state distribution of the endolysosomal system in the axon, dendrites, and cell body. Degradative lysosomes are predominantly located in the cell body , represeEarly work measuring the pH of axonal endocytic cargos demonstrated an increasing frequency of acidic organelles as cargos moved from distal to proximal , suggestL-phenylalanine 2-naphthylamide  in neurons showed enriched hCTSD puncta in somatodendritic regions of layer 5 cortical neurons and hippocampal CA1 neurons . AlthougNeurons require efficient transport mechanisms to maintain a steady-state distribution of endolysosomes throughout the cell. Kinesin and dynein motors drive long-distance transport along microtubule (MT) tracks in axons and dendrites, whereas myosin motors mediate short-range movement along actin filaments enriched in growth cones and synaptic regions . BidirecBorcs7 (encoding a BORC subunit) develop progressive axonal dystrophy and motor dysfunction, indicating a critical role of BORC-dependent axonal lysosome transport for maintaining axon integrity and neuron function in vivo  mouse dystrophic axons contain immature lysosomes with low levels of proteases, representing a population of lysosomal intermediates distinct from mature lysosomes in the cell body  and thus facilitates retrograde LE transport from axonal terminals toward the soma . Deletinradation  and enharadation . Interesradation  that is radation , althougGlued  transport and positioning responds to alterations in cytoplasmic pH. Acidification of the cytosol causes lysosome dispersal into neuronal processes, whereas cytosolic alkalization shifts LE/LY distribution to the cell body , suggestDegradation of autophagic cargos depends on their dynamic interactions with endolysosomes, which supply the hydrolytic enzymes needed for substrate degradation . Both thIn axons, autophagosomes predominantly form at distal terminals , then suThus, the maturation of these distal AVs into autolysosomes is tightly linked to their retrograde transport. Scaffolding proteins, including JIP1, HTT-associated protein 1 (HAP1), and JIP3 , regulatAlthough dynein-driven retrograde transport of autophagosomes was suggested, a fundamental question remained as to how autophagosomes generated at distal axons acquire dynein motors for retrograde transport toward the soma. Using live rat dorsal root ganglion neurons combined with molecular disruption of autophagosome fusion with LEs or impairment of dynein–snapin (motor–adaptor) coupling, a study revealed a motor-adapter sharing model in which LE-loaded dynein–snapin complexes are shared by autophagosomes upon their fusion . BlockinRecent evidence further revealed the role of lysosomes in axonal autophagosome maturation and clearance. Lysosomes that transport from the soma into axons target autophagic cargo to facilitate local AV maturation and cargo degradation. These lysosomes contain active lysosomal enzymes and are continuously delivered to distal axons from the cell body . DisruptLysosome dysfunction and impaired trafficking are linked to the pathogenesis of rare and common neurodegenerative diseases, including frontotemporal lobar degeneration (FTLD), Alzheimer’s disease (AD), amyotrophic lateral sclerosis (ALS), Huntington’s disease (HD), Parkinson’s disease (PD), and lysosomal storage disorders (LSDs) such as Niemann-Pick disease type C NPC; . In thesTMEM106B, encoding a transmembrane protein primarily localized on LEs and lysosomes, have been linked to several neurodegenerative diseases, including FTLD  interacts with DIC  leads to enhanced recruitment of JIP4 to AV membranes, inducing abnormal activation of the anterograde motor kinesin  be differentially regulated in response to certain stimuli and signaling pathways; (2) associate with unique endolysosome subpopulations at different maturation stages or with specific functions; (3) selectively target to organelles within different segments of long axons; (4) play cell type–specific roles depending on their expression levels within a given neuronal subtype; or (5) serve overlapping or partially redundant functions. Therefore, further work is needed to examine these possibilities, which will conceptually advance our understanding of how these motor–adaptor transport complexes are regulated and coordinated to sustain efficient clearance capacity in distal regions under physiological conditions and become disrupted under pathological conditions.Moreover, many other important issues also need to be addressed in future studies. For example, do neuronal lysosomes in the periphery or distal regions serve as environmental or metabolic sensors to activate transcriptional pathways in the soma and promote lysosomal adaptation to stress? What are the upstream signaling events that regulate neuronal lysosome trafficking within axons and positioning at sites of high axonostatic demand or under autophagic stress? What role do cytoskeletal modifications play in modulating axonal lysosomal movement? How do interactions with other organelles such as the ER or mitochondria contribute to lysosome positioning and local degradation capacity in axons? Do these dynamic events change throughout development and aging? Addressing these questions will help advance our understanding of neuronal endolysosomal transport and lysosomal functionality under physiological conditions and how impaired trafficking and lysosomal dysfunction contribute to neurodegenerative diseases. Understanding the disease-linked mechanisms underlying axonostatic failure is critical for designing potential therapeutic strategies that target axonostatic restoration to support neuronal survival and function.


Theories suggest that with increasing age, adults more effectively regulate their emotions and seek to limit high physiological arousal. Prior research indicates physical activity attenuates negative affect reactivity to stress, but also increases physiological arousal. The present study extends prior work by examining age-related differences and changes over time in the extent of attenuation. Participants , from the National Study of Daily Experiences completed 8 end-of-day assessments of their negative emotions, stress, and physical activity across 3 measurement bursts spaced approximately 10 years apart. Results from three-level multilevel models suggest that when full random effects are specified, physical activity does not attenuate negative affect reactivity to stress. Additionally, extent of attenuation did not differ with age or change over time. Discussion pertains to how these findings advance theoretical understanding of socioemotional development and to methodological nuances of random effects and non-normally distributed data.


Prior studies of caregiving characteristics by type of caregivers are inconsistent, particularly those who are spouses and adult children. This study examined caregiving characteristics between spouses and adult children of cognitively impaired older adults. We analyzed phone-screening data from an ongoing trial of a dyadic sleep intervention program for persons with dementia and their caregivers. Data included spouse caregivers (n=52) and adult child caregivers (n=24). Nearly all participants (95%) lived with their care recipients (91% with dementia). Types of caregiving activities were measured by activities of daily living [ADLs] and instrumental ADLs [IADLs] with their levels of intensity . Care recipients’ sleep was measured by the Neuropsychiatric Inventory-Nighttime Behavioral Subscale (8 items). Analyses included Pearson correlations and t-tests. Adult child caregivers helped their care recipients at significantly higher levels as indicated by their measure of dependence in dressing , continence , laundry , and transportation  than spouse caregivers. Adult child caregivers also reported their care recipients having more difficulty falling asleep  and having more numbers of sleep problems than spouse caregivers . The findings suggest that adult child caregivers may involve higher levels of caregiving responsibilities during daytime and nighttime, compared to spouse caregivers. Further research needs to explore complimentary ways to involve spouse and adult child caregivers in the care of this vulnerable population.


ABSTRACT IMPACT: This study advances our understanding of potentially key drivers in the early formation of pancreatic cancer, a disease with few treatment options and poor patient outcomes. OBJECTIVES/GOALS: Patients diagnosed with pancreatic ductal adenocarcinoma (PDAC) have a 5-year survival rate of ˜9%. A key driver of poor patient outcomes is late-stage diagnosis. A better understanding of PDAC onset is needed. This study was developed to understand how extracellular vesicles may be involved in the early formation of PDAC. METHODS/STUDY POPULATION: Extracellular vesicles (EVs) were isolated from several human PDAC and normal pancreatic cell lines, using ultracentrifugation with filtration or size exclusion chromatography. We next treated normal pancreatic cell lines with cancer cell EVs (cEVs). Next generation sequencing was used to measure global gene expression changes after treatment. Validations were performed using qPCR and luciferase activity assays. Multi-omics characterization of EVs was accomplished using mass spectrometry based proteomics, metabolomics and lipidomics analysis. RESULTS/ANTICIPATED RESULTS: We found that normal cells upregulated a variety of stress response pathways in response to cEVs. Lipid synthesis was also severely downregulated in these cells. We further validated activation of the unfolded protein response (UPR) in normal cells treated with cEVs. Multi-omics characterization of cEVs identified several enriched proteins, lipids and metabolites which may play a role in the activation of the UPR. DISCUSSION/SIGNIFICANCE OF FINDINGS: Our results indicate that cEVs induce stress, and in particular the UPR, in normal pancreatic cells. Long-term UPR can impact a variety of cancer hallmarks. The UPR can mediate progression of pancreatic intraepithelial neoplasia (PanIN) to PDAC. Our results highlight a potential role for cEVs to alter the function of normal cells, aiding disease onset.


Few studies have explored the underlying pathways between environment cognition  and mental health in older adults. We tested the mediation effects of physical activity and place attachment in the relationship between environmental cognition and mental health, based on a survey study of 1,553 older adults in Hong Kong using structural equation model. The results showed that significant relationship between negative environmental cognition  on access to convenient stores, leisure facilities, clinics, community centers, religious places and lower mental health can be explained by lower daily average physical activity time. Place attachment can significantly mediate the positive effect of positive environmental cognition  towards all types of services on mental health. Findings from this study have policy implication for urban planning and age-friendly community design


Studies have demonstrated a link between sensory impairment and dementia risk, but little is known about the presence of beta-amyloid plaques in individuals with single and multisensory impairments. Sensory function  and amyloid PET imaging were measured in 170 BLSA participants  from 2012 to 2019. Log-binomial regression models were used to examine the prevalence ratios (PR) of amyloid positivity for individual sensory impairments and across categories of impairments. While crude associations indicate associations of vision impairment  and impairments in all four senses  with amyloid positivity, these associations were insignificant after adjusting for age, sex, race, and education. There were no other crude and adjusted associations. These results suggest sensory impairments may be related to dementia independent of AD pathology. Future studies with larger sample sizes are warranted.


Aging is the most prominent risk factor for cognitive decline, yet behavioral symptomology and underlying neurobiology can vary between individuals. Certain individuals exhibit significant age-related cognitive impairments, while others maintain intact cognitive functioning with only minimal decline. Recent developments in genomic, proteomic, and functional imaging approaches have provided insights into the molecular and cellular substrates of cognitive decline in age-related neuropathologies. Despite the emergence of novel tools, accurately and reliably predicting longitudinal cognitive trajectories and improving functional outcomes for the elderly remains a major challenge. One promising approach has been the use of exosomes, a subgroup of extracellular vesicles that regulate intercellular communication and are easily accessible compared to other approaches. In the current review, we highlight recent findings which illustrate how the analysis of exosomes can improve our understanding of the underlying neurobiological mechanisms that contribute to cognitive variation in aging. Specifically, we focus on exosome-mediated regulation of miRNAs, neuroinflammation, and aggregate-prone proteins. In addition, we discuss how exosomes might be used to enhance individual patient outcomes by serving as reliable biomarkers of cognitive decline and as nanocarriers to deliver therapeutic agents to the brain in neurodegenerative conditions. Age-related cognitive decline remains a prominent public health concern, with Alzheimer’s Disease (AD) accounting for the majority of dementia diagnoses  . DespiteAging is the most prominent risk factor for cognitive decline, but behavioral symptomology and underlying neurobiology can vary between individuals. Indeed, some individuals exhibit significant cognitive impairments in comparison to age-matched controls, while others maintain intact cognitive functioning or display only minimal decline . AlthougAPOE, APP, PSEN1 etc.) as well as rare  genetic variants  whose primary function is to translocate biological substrates  between cells . Exosomethe cell . While tthe cell . Of notethe cell . Regardithe cell .in vitro supernatant vs. clinical plasma/serum), processing generally involves isolation  and characterization  followed by biological interpretation of exosomal quantity and contents utilizing complementary biochemical techniques    . Renewedys etc.) . Given tys etc.) . The capys etc.) . It shouys etc.) ; howeverys etc.) .Attempts have been made to categorize exosomes according to their net advantages or disadvantages for CNS homeostasis, yet such a dichotomous distinction may be misleading. For instance, accumulating evidence illustrates exosomes facilitate symbiotic dynamism between neurons and oligodendrocytes, whereby activity dependent synaptic transmission triggers exosomal secretion from oligodendrocytes that contain essential myelin components ; subsequently, internalization of these exosomes amongst proximal neurons enhances cellular viability . ConversA large quantity of exosome research has focused on their capacity to incorporate and transport microRNAs (miRNAs). These short  non-coding RNAs function as post-transcriptional regulators of gene expression by binding to complementary pairing sites on mRNA  across various locations within a cell, including the nucleus, cytoplasm and subcellular compartments, such as stress granules . Such biin silico analysis linked these miRNAs to stress response and neurotrophic signaling pathways, findings suggested miRNA mediated regulation of these biological processes may increase risk for cognitive decline . While a large set of miRNAs showed significant positive correlations with age, another set maintained significant negative correlations with cognitive performance, including those miRNAs selectively expressed in the brain. In turn, functional annotation of miRNAs associated with poor performance revealed several biological pathways, including neurotrophin signaling, whose miRNA-mediated regulation may be important mediators of cognitive decline . Further decline . Interes decline .To gain more reliable insights into the neurobiological mechanisms of cognitive decline, additional investigations have interrogated the role of miRNAs from brain derived exosomes. After isolating neural exosomes from AD plasma samples, researchers recently identified a set of significantly upregulated  and downregulated  miRNAs which were predicted to regulate several homeostasis pathways in the CNS, including steroid biosynthesis and mTOR signaling . In anotin vivo delivery via exosomes was capable of ameliorating memory deficits in rodents  otherwise induced by intracerebral application of Aβ oligomers; furthermore, researchers inferred these benefits may be attributed to the increased synaptic phosphorylation of calmodulin-dependent kinase II   whose inase II . Similarpathway) .To expand our understanding for the role of exosomal miRNAs in mediating the underlying mechanisms of cognitive decline, several lines of inquiry require elucidation, some of which are addressed below. For one, we currently lack a comprehensive understanding of the mechanisms within recipient cells that are modulated by exosomal miRNAs. Previous evidence suggests miRNAs released by exosomes remain functional in their recipient cells, capable of directly downregulating the expression of target transcripts, similar to endogenous effects in parent cells . Interesinflamm-aging, is a common feature of normal aging. Higher levels of neuroinflammation are a risk factor for age-related cognitive decline and the accumulation of neuropathological hallmarks , which ultimately resulted in the disruption of plasma membrane integrity as well as elevated neurotoxicity ; increase anti-inflammatory cytokines ]; in turn, this is associated with the mitigation of pathological peptides  and the preservation memory capacities . Interestingly, such beneficial effects on cognition and neuroinflammation were comparable when exosomes were specifically targeted to the brain or administered systemically, highlighting the capacity of these EVs to exert their effects across the BBB , 2019. FTo enhance our understanding of exosomal-mediated neuroinflammation and associated risk for cognitive decline, several lines of evidence require further interrogation. Most prominently, further studies are needed to characterize the contents of exosomes which are responsible for alterations in CNS inflammation. Along with pathogens , a variety of molecules can induce immune responses in the brain, including reactive oxygen species (ROS), purine metabolites , calcium-binding proteins and lipotoxic ceramides . In addiEvidence indicating the association between aggregate-prone proteins  and age-related cognitive decline is abundant, with increasing data suggesting their spread between CNS cells can be facilitated through exosomes . It shou1–42, pTau-181 and pTau-S396 in neuronal exosomes isolated from blood samples could differentiate AD patients from controls, as well as those patients with MCI who remained cognitively stable from those who illustrated progressively declining MMSE scores. Furthermore, intracerebral injection of such exosomes into healthy adult mice induced a significant accumulation of pTau in CA1 pyramidal neurons, compared to those exosomes from stable MCI participants , individuals who illustrated higher exosomal Aβ1–42 levels maintain an 11.1- and 8.5-fold increased risk for significant cognitive deterioration at 2 and 3 year follow-up assessments, respectively, compared to MCI participants who remained cognitively stable  via exosomes is capable of increasing translation of these proteins in recipient cells; such exploitation of host cell gene expression machinery could facilitate the accumulation of protein aggregates in otherwise healthy CNS cells  may synergistically compromise CNS homeostasis . Such a NS cells . AnotherNS cells . FurtherPredicting individual cognitive trajectories in aging can be important for diagnostic purposes, and the adoption of exosomes as biomarkers may prove particularly useful in this respect. Clinical assessment alone can fail to discriminate those individuals who maintain cognitive resilience from those who develop MCI, as well as those MCI patients who go on to develop dementia . AlthougNumerous studies have used miRNAs isolated from blood-derived exosomes as biomarkers to classify individuals who are exhibiting late life cognitive dysfunction. Compared to blood samples, available evidence suggests measures of exosomal miRNAs obtained from CSF samples offer similar accuracies in distinguishing individuals with age-related cognitive impairment . HoweverAlthough there is no consensus on which individual miRNAs or groups of miRNAs from exosomes best predict cognitive impairment, some consistent evidence has emerged. For example, levels of miR-342-3p in plasma exosomes were significantly lower in AD participants compared to controls and correlated with poor cognitive performance  among elderly, cognitively unimpaired individuals in a separate study . SimilarIn addition to cross-sectional investigations, several studies have used exosomes as biomarkers to predict longitudinal variation in cognitive decline. Using neurally derived exosomal concentrations of aggregate-prone peptides, researchers can predict individuals who progress from MCI to AD-dementia within 36 months , as well as cognitively sound individuals who develop dementia up to 10 years later  . IncreasTo enhance the utilization of exosomes as reliable and valid biomarkers, several important limitations should be considered. When examining their associations with cognitive decline, researchers should take steps to differentiate the tissue-specific  as well as the cell-specific origin of exosomes. Indeed, data indicate exosomes from the CNS contain significantly different molecular contents depending on an individual’s cognitive capacities  as well as their cellular origin , while such differences in content can subsequently induce distinct functional consequences for recipient cells . InvestiDespite limitations, the implementation of exosomes as biomarkers for age-related cognitive decline offers several advantages. As mentioned previously, exosomal sampling is more cost effective and less invasive than currently available biomarkers . Furthermore, exosomes derived from the CSF and blood offer similar accuracy for identifying individuals exhibiting cognitive decline, while neuroimaging and blood derived exosomes exhibit similar specificity for distinguishing dementia cases from healthy controls . Meanwhiin vitro   . For inseatment) . Researceatment) . Similareatment) .Perhaps the greatest advantage offered by exosomes is their capacity to transverse the BBB due to their small size, endogenous biological properties, and low immunogenicity, which has otherwise been a major obstacle in treatment development for cognitive decline . This ha1–40, Aβ1–42, plaques)  can reliably improve performance on cognitive tasks  and inhibit biological processes associated with cognitive impairment, including the dysregulated expression of pro- and anti-inflammatory cytokines (IL1-β and IL-10), the activation of CNS immune cells  and the deposition of Aβ (Aβplaques) . Additioplaques) . Togetheplaques) .1.While the clinical development of therapeutic interventions faces a variety of hurdles, the consideration of exosomes as a novel delivery vehicle for future nanotherapeutics may prove to enhance outcomes for those individuals vulnerable to age-related cognitive pathologies. In this context, the use of un-modified exosomes isolated from adipose mesenchymal stem cells is currently being investigated for AD-dementia in clinics (NCT04388982). Although the study is only exploring the safety and efficacy of exosome treatment in patients, data from secondary outcome measures that include AD biomarkers based in Aβ plasma/CSF measures, PET scans, and neuropsychological tests, may further encourage the development of engineered vesicles for the treatment of age-related cognitive decline. The current composition proposes exosomes are uniquely suited to improve our mechanistic understanding as well as enhance patient outcomes for the growing public health challenge of age-related cognitive decline. We highlight how the analysis of exosomes can improve our understanding of varying mechanisms in age-related cognitive decline, including the role of miRNAs, neuroinflammation and aggregate-prone proteins . Despite these contributions, we suggest specific areas of inquiry that warrant further interrogation, including the elucidation mechanisms in recipient cells that are modulated by exosomal miRNAs, the characterization of exosomal contents that are responsible for alterations in neuroinflammation and the examination of other aggregate-prone peptides  in exosomes that may synergistically increase risk for late life cognitive impairment. Moreover, we discuss how exosomes might be used to enhance outcomes for elderly individuals by serving as reliable biomarkers and therapeutic agents.To improve the reliability of studies going forward, consistent nomenclature coupled with protocol modifications that enhance the discrimination of EVs could be particularly effective. Due to similar biophysical properties , isolation techniques developed for exosomes can inadvertently lead to results that are derived from samples containing a variety of different EVs . SimilarSeveral additional limitations hinder the further application of exosomes discussed herein. Notably, it remains unknown whether concurrent variation in exosomes and cognitive capacities is reflective of a causal or reverse-causal association. In other words, it is difficult to determine if exosomal differentiation contributes to divergent cognitive trajectories, or if such differences are secondary to other biological processes that otherwise increase risk for cognitive deterioration in aging. A lack of data measuring time-dependent variation in the profiles of neuronal exosomes contributes to this lack of understanding, and potentially inhibits researchers from adequately controlling for the effects of aging. Although a limited set of investigations have employed longitudinal designs , future MD and VP conceived and designed the study. MD and AL conducted the literature search. MD, AL, and VP wrote the manuscript. TF and MW edited the final manuscript. All authors have read and approved the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


One of these involve a de novo heterozygous variant in an X-linked gene (ARHGEF9) in a female individual. We hypothesize the skewed X-inactivation may result in primarily expression of the pathogenic variant. We anticipate identifying additional candidate variants in coding regions of genes previously not associated with EMAS or pediatric epilepsies as well as in noncoding regions of the genome. DISCUSSION/SIGNIFICANCE OF IMPACT: Despite the genetic heterogeneity of EMAS, our initial analysis identified de novo pathogenic or likely pathogenic variants in 15% (6/40) of our cohort. As the cost continues to decline, short read genome sequencing represents a promising diagnostic tool for EMAS and other pediatric onset epilepsy syndromes. CONFLICT OF INTEREST DESCRIPTION: The authors have no conflicts of interest to disclose. SD has consulted for Upsher-Smith, Biomarin and Neurogene on an unrelated subject matter. GLC holds a research collaborative grant with Stoke therapeutics on unrelated subject matter.OBJECTIVES/GOALS: Epilepsy with myoclonic-atonic seizures (EMAS) is a childhood onset epilepsy disorder characterized by seizures with sudden loss of posture, or drop seizures. Our objective was to use short-read genome sequencing in 40 EMAS trios to better understand variants contributing to the development of EMAS. METHODS/STUDY POPULATION: Eligibility for the cohort included a potential diagnosis of EMAS by child neurology faculty at Children’s Hospital Colorado. Exclusion criteria included lack of drop seizures upon chart review or structural abnormality on MRI. Some individuals had prior genetic testing and priority for genome sequencing was given to individuals without clear genetic diagnosis based on previous testing. We analyzed single nucleotide variants (SNVs), small insertions and deletions (INDELs), and larger structural variants (SVs) from trio genomes and determined those that were likely contributory based on standardized American College of Medical Genetics (ACMG) criteria. RESULTS/ANTICIPATED RESULTS: Our initial analysis focused on variants in coding regions of known epilepsy-associated genes. We identified pathogenic or likely pathogenic variants in 6 different individuals involving 6 unique genes. Of these, 5 are


Teaching point: The probable mechanism of pseudoaneurysm formation related to metastatic neoplasm is a tumor embolus penetrating and destroying the vessel wall.Pulmonary artery pseudoaneurysm (PAP) related to metastatic neoplasm is rare. We describe a unique case of multiple PAPs secondary to metastatic uterine leiomyosarcoma and demonstrate the serial chest computed tomography to support the theory that the tumor begins as a tumor embolus, followed by infiltration and breakdown of the vessel wall, leading to aneurysmal dilatation and invading the perivascular tissue. Figure 1). Computed tomography pulmonary angiography (CTPA) was performed to rule out acute pulmonary thromboembolism and showed innumerable heterogeneously enhanced masses scattered in both lungs, surrounding segmental branches of pulmonary arteries. Multiple fusiform pulmonary artery pseudoaneurysms surrounded by soft tissue masses were also noted . A review of serial chest computed tomography (CT) revealed tumor emboli in the peripheral branches of pulmonary arteries , which finally developed into pseudoaneurysms with surrounding soft tissue masses .A 49-year-old female with known uterine leiomyosarcoma and lung metastasis with multiple tumor thrombi in pulmonary arteries was receiving palliative chemotherapy. Prior to her fourth chemotherapy session, the patient developed acute-onset dyspnea. Chest radiograph revealed a larger size of the multiple well-defined pulmonary masses scattered in both lungs; therefore, progressive pulmonary metastasis was suspected . The serial chest CT also showed gradual dilatation of peripheral branches of right pulmonary arteries ; this could be due to tumor growth and continued destruction of vessel walls.A similar mechanism of pathogenesis is thought to have occurred in this case. From the serial chest CT, the PAPs with surrounding soft tissue masses had previously shown intraluminal thrombus and dilated pulmonary artery, which resembled tumor thrombus (Abnormal dilatation of the pulmonary arteries and veins was also observed in the patient’s CTPA with similar findings previously described in a reported case of metastatic uterine sarcoma . The onlIn conclusion, PAPs caused by metastatic neoplasm may firstly begin as tumor emboli, later progressing into pseudoaneurysms as we elucidated in the serial chest CTs and CTPA findings in this case.


Previous studies have shown that engaging in musical activities throughout the lifespan may buffer age-related decline in auditory and motor function, as well as in general cognitive function. MRI studies have demonstrated that individuals with musical training and experience exhibited greater grey matter volume and functional connectivity in extensive brain regions, especially in auditory and motor systems, compared to matched controls with no particular musical training or experience. Therefore, musical activity is a potential protective factor for brain health across lifespan. However, how lifespan musical experience shapes functional connectivity in older adults is still unknown. The current analysis investigated whether general musical experience (Goldsmith Music Sophistication Index) is associated with functional connectivity in older adults , focusing on seed regions in primary motor areas  and primary auditory regions  and their functional connectivity towards other areas throughout the whole brain. We found that older adults with more musical experience showed greater functional connectivity between anterior superior temporal gyrus and insula , and between posterior superior temporal gyrus and cerebellum . However, musical experience and music-related functional connectivity was not significantly correlated with general cognitive functions in our sample. Overall, our findings suggest that older adults with more musical experience might be more efficient in some aspects of auditory processing and auditory-motor skills, but this may not transfer towards domain-general cognitive tests. Our results support the notion that even non-professional engagement in musical experiences may afford benefits to the aging brain.


Dear Editor,we read with great interest the study by Vetrugno et al.  where thWe propose a brief case to suggest taking Inferior Vena Cava Filter (IVCF) placement into account as a prevention tool for recurrent fat embolism (FE) in selected patients. A 16-years old male was involved into a motor-scooter accident reporting right femoral shaft fracture and left tibia shaft fracture. External fixation was provided as a bridge treatment to surgery. A few hours later he suddenly developed respiratory failure and coma and was admitted to our Intensive Care Unit (ICU) for supportive care. Cerebral Computed Tomography (CT) was normal; total body CT scan with contrast excluded pulmonary thromboembolism (PTE) and showed diffuse interstitial and alveolar edema; electroencephalogram identified status epilepticus for which antiepileptic drugs were started. Cerebral magnetic resonance imaging revealed a “starfield” pattern compatible with the diagnosis of cerebral fat embolism. Patent foramen ovale was excluded. He was treated conservatively with mechanical ventilation with gradual improvement of his respiratory status, whereas neurologic impairment needed prolonged sedation. During his stay in the ICU he underwent sudden unexplained respiratory distress episodes, suggesting that embolization was possibly recurring. When stable, on day 7 of hospitalization, the patient was scheduled for surgical treatment. An IVCF was placed the day prior to surgery with the aim of stopping more fat emboli from spreading during intramedullary nailing of femur and tibia (Fig. Although IVCF insertion is discouraged for primary prevention of PTE in both orthopaedic surgery  and majoDespite scarce evidence, we reckon that IVCF could improve prognosis in selected patients with FES, especially those presenting high risk factors, after accurately weighing risks and benefits. This measure may also help to earlier stabilize the patient so that definitive surgical treatment of the fractures is promptly provided.In conclusion, the study by Vetrugno et al. shed light on risk factors related to FES. However, further research is necessary to identify the best treatment and prevention strategies. Structured studies and prospective evaluation are needed to establish the effective benefits of IVC filter placement in similar cases.


Given the frequent overlap between biological plant invasion and ecological restoration efforts it is important to investigate their interactions to sustain desirable plant communities and modify long-term legacies both above- and below-ground. To address this relationship, we used natural reference, invaded and created vernal pools in the Central Valley of California to examine potential changes in direct and indirect plant effects on soils associated with biological invasion and active restoration ecosystem disturbances. Our results showed that through a shift in vegetation composition and changes in the plant community tissue chemistry, invasion by non-native plant species has the potential to transform plant inputs to soils in vernal pool systems. In particular, we found that while invasive plant litter decomposition was driven by seasonal and interannual variability, associated with changes in precipitation, the overall decomposition rates for invasive litter was drastically lower than native species. This shift has important implications for long-term alterations in plant-based inputs to soils in an amplifying feedback to nutrient cycling. Moreover, these results were independent of historic active restoration efforts. Despite the consistent shift in plant litter decomposition rates and community composition, we did not detect associated shifts in below-ground function associated with invasion by non-native plants. Instead, soil C:N ratios and microbial biomass did not differ between invaded and naturally occurring reference pools but were reduced in the manipulated created pools independent of invasion levels. Our results suggest that while there is an observed invasive amplifying feedback above-ground this trajectory is not represented below-ground, and restoration legacies dominated 10 years after practices were applied. Restoration practices that limit invasive plant feedbacks and account for soil legacy recovery, therefore offer the best solution for disturbed ephemeral ecosystems. Applications of restoration practices in wetland systems frequently must contend with plant invasion by non-targeted species, and these invading plants can undermine restoration goals through impacts both above- and below-ground. In this study we examined the impacts of invasive plants among natural and created vernal pools in California and found that invasion had a large effect on the plants present in the community and especially influenced plant litter decomposition rates. These invasive species caused changes can have implications for long-term alterations in plant-based inputs to soils; however, we did not detect associated shifts in below-ground function associated with invasion by non-native plants. The vegetation community has a key role in influencing soil properties through increased soil physical stabilization , alterinThe impact of changes in plant–soil feedbacks associated with plant invasion on ecosystem processes is varied  as well In restoration ecology, a common response to increased threat from invasive species is to implement management strategies to reduce the prevalence and spread of the incoming plant species ; howeverseeVernal pools are shallow ephemeral wetlands found in flat to low slope grasslands with poorly draining soil that facilitates ponding. They are predominately precipitation-fed and defined by abrupt edges delimited by locations of ponding . HistoriTo examine the consequences of invasion in the context of restoration through plant–soil feedbacks on ecosystem processes, we examined a series of natural reference pools dominated by native species, naturally occurring pools invaded by non-native plants and created vernal pools dominated by non-native plants all located in a grassland site in central California. Among these contrasting restoration and invasion legacies we asked three primary questions: (i) How does invasion and restoration in vernal pool systems influence above-ground litter decomposition? (ii) How might shifts in above-ground community composition impact litter inputs to soils? (iii) Have shifts in above-ground community composition prompted changes in below-ground ecosystem function? Within this framework we hypothesize that ecosystem processes being driven by the above-ground presence of invasive plants will increase both carbon and nitrogen into the soil through altered plant litter and decomposition rates causing positive feedbacks, yet the plant material chemical ratios will favour increased nitrogen and thus increase microbial biomass in the invaded sites regardless of restoration status. Alternatively, active restoration, such as creating a disturbance of scraping the topsoil layer, has the potential to expose new soil layers lowering overall carbon and nitrogen content and increasing C:N ratios, pH and altering soil texture thereby producing lasting effects below-ground that could potentially overshadow the impacts of biotic invasion.seeseeWe conducted our experiment in the Central Valley of Solano County, CA, USA . This region experiences a strong seasonality, with most precipitation falling during the winter months between November and February, a mean annual temperature of 20.1 °C, and mean annual rainfall of 500 mm . In our study site located at the Travis Air Force Base, Solano County, CA, both naturally occurring ‘reference’ vernal pools, that have existed in a single grassland meadow for decades, as well as pools created for restoration purposes are present across the 15-ha study site. This meadow contains a network of pool basins and upland grassland vegetation representative of regional vernal pool and grassland systems see. To examseeOver time (approximately in years 2006–08), a subset of the naturally occurring pools in this landscape also became dominated by invasive non-native annual grasses (hereafter ‘invasive’) while other pools eluded invasion, maintaining native species see. Given tWe also assessed variation in the chemistry of plant tissues from 18 vernal pool species as well as upland dominant species common to the site and surrounding grassland communities, to examine community-scale changes in stoichiometry and tissue chemistry from individual plants distributed across the site. These metrics were selected to target potential plant inputs on nutrient and carbon cycling among the three different vernal pool types. Specifically, we measured plant tissue concentrations of nitrogen (N), cellulose, lignin and carbon (C), and calculated the associated ratios between these components carbon:nitrogen (C:N) and lignin:nitrogen (L:N). All plant species were collected during peak biomass in 2011 coinciding with species composition surveys as measurement of growing season standing pools of plant chemistry. Cellulose and lignin, which are important components in litter decomposition and photodegradation . Plant Cin situ above-ground plant decomposition rates through the utilization of litter bags. While using vegetation from peak biomass prevents the potential for nutrient translocation and resorption and consequently influences tissue chemistry, plant inputs in this system do not commonly undergo full senescence prior to mowing and deposition on the landscape, similar to N = 72 litter bags above field-collected invasive litter and N = 72 litter bags below). While 1 cm of litter has been shown to impact vernal pool vegetation structure ; thus, we used recently cut foliage, collected from both upland and pool areas across the larger research site, for key species during spring peak biomass the season prior to field implementation to determine tructure , placingTo examine environmental and biotic drivers of vernal pool litter decomposition we installed litter bags for two winter wet seasons during the period when ponding is expected and one summer dry season, as well as comparing species effects on decomposition for the first wet season. We installed ‘wet season’ litter bags in the fall prior to any substantial precipitation (2011–12 and 2012–13) and removed them in the spring after pools dried down and the annual vegetation had bolted. These field incubations for the winter wet decomposition studies lasted from September to April. We placed the ‘dry season’ litter bags (2013) in the field just after spring drydown and removed them before any substantial autumn rains (April–September).Lolium multiflorum, an non-native invasive annual grass species that is ubiquitous in both created and invaded reference pools at the study site , only the invasive grass L. multiflorum was used in the sequential litter bag deployments (wet season 2012–13 and dry season 2013) due to low site-level abundance of the native grass P. californicus.For the first year of the study, we were able to include litter bags for both the dominant native and invasive species for the wet season 2011–12 sampling period to test for species-level non-native invasive versus native decomposition differences. The invasive species litter was composed of pure udy site . The natN = 72 at 125 cm3 each) were placed on dry ice immediately upon sampling. Samples were kept cold at 4 °C until processed, with the majority of soil analyses  completed within 1 month of field collection .We collected bulk soil samples during peak flowering and peak above-ground plant biomass (April) in 2011 as the annual plants are dependent on soil processes while active during this brief period of highest productivity. Three soil samples were collected per pool, with one sample at each of three elevational plot locations within each pool . Soil samples (We obtained soil moisture through the gravimetric soil moisture method (% soil moisture = 100 * (fresh weight − dry weight)/dry weight). We measured soil pH using a 1:3 soil to water ratio  on soil subsamples, where the pH subsample was measured three times to calculate a mean value for each elevational location within each pool. We used a subset of the pool bottom soils to obtain soil texture  using a modified version of For all univariate data analysis, we used linear mixed-effects models in R version 4.0.0  in the lFor our multivariate analyses, we examined differences among pool types based on community-scale chemistry traits, including tissue C, N, lignin and cellulose. Community composition was used to scale chemistry traits based on abundance of dominant plant species present in vegetation plots during the same year as tissue sample collection for chemical analysis (2011). We created species traits matrices based on multiplying species counts by species-level chemical traits, thereby creating four independent species trait matrices scaled by species presence, in addition to the community composition matrix of species counts that included all species present in each plot. These trait and composition matrices were examined for differences among pool types using a permutational multivariate analysis of variance (perMANOVA) test based on a Bray–Curtis dissimilarity matrix (R package Adonis). Multiple comparisons among pool types were conducted using the function pairwiseAdonis, which accounts for multiple comparisons and uses a Bonferroni correction . We applF2, 69 = 4.9, P < 0.01) with different compositions in reference pools from pools created through creation  or affected by invasive species , N , lignin  and cellulose , suggesting differences in the drivers of decomposition linked with invasion.Finally, decomposition differed substantially associated with species selection  where the created pools (C:N-10.7) were significantly lower than the reference (C:N-11.7) and invaded pools (C:N-11.8). We found that soil microbial biomass C when compared across pool types also varied . The lowest average microbial biomass was found in the created pools and was significantly lower than the reference pools based on effect size.Many of the fundamental physical soil properties we measured did not differ among vernal pool types associated with invasion or creation ; howeverThe introduction of invasive species can alter the ecosystem through a variety of pathways creating amplifying feedbacks that exacerbate further ecosystem changes . To combseePhragmites australis, however, shows a similar decrease in decomposition rates driven by higher C:N ratios and higher concentrations of lignin ultimately promoting nutrient competition for native plant species (Invasive species can alter ecosystems through a wide variety of pathways, ranging from shifts in community interactions to changes in nutrient cycling and physical ecosystem properties  and this species .While we observed that invasive non-native decomposition was slower than native species in our study, an alternate positive feedback to soil nutrients associated with invasive effect on the ecosystem has been noted in this vernal pool system . Here thLong-term averages of litter decomposition are driven largely by climate and microbial biomass  and the Restoration ecology focused on evaluating restoration success in depressional wetlands more generally has suggested that the nutrient composition of vegetation is an important metric in evaluating long-term restoration success .Differences in plant community composition, tissue chemistry and decomposition rates can have a cascade of effects on the below-ground, such as speed of nutrient cycling, microbial community composition (While a direct effect of a restoration legacy was observed below-ground in soil C:N and generally in extracted microbial biomass C, not all soil metrics tested demonstrated this artefact of restoration nor demonstrated a change solely due to invasion, in contrast to studies comparing reference and restored pools with greater differences in vegetation community structure (i.e. woody plants; Similar interacting processes likely influenced our findings for differences in soil pH among pool types that were not connected to either invasion or restoration independently. Invasive plants have been shown to alter the soil pH, both increasing and decreasing depending on the invasion type and magnitude , and we In summary, despite strong changes in above-ground plant contributions the soil metrics directly linked to plant inputs did not show the same response to the plant invasion. Instead, organic soil components exhibited a legacy of vernal pool active restoration from the decade prior. This study demonstrates that while above-ground vegetation may vary strongly in its abundance and composition and produce conditions that facilitate further invasion, this signal does not necessarily translate below-ground even after multiple years of invasion. The success of a restoration project is often based solely on the above-ground vegetation, and in this regard, further restoration efforts to minimize invasive species presence is required for our vernal pools. However, this study illustrates an opportunity to limit long-term effects of altered above-ground plant–soil feedbacks on below-ground processes should additional restoration occur. The importance of this time lag in establishing altered community feedbacks within the ecosystem provides a window of opportunity for adaptive management practices aimed at ecosystem recovery.The following additional information is available in the online version of this article—plab042_suppl_Supplementary_Figure_S1Click here for additional data file.


ABSTRACT IMPACT: Identifying factors associated with opioid overdoses will enable better resource allocation in communities most impacted by the overdose epidemic. OBJECTIVES/GOALS: Opioid overdoses often occur in hotspots identified by geographic and temporal trends. This study uses principles of community engaged research to identify neighborhood and community-level factors associated with opioid overdose within overdose hotspots which can be targets for novel intervention design. METHODS/STUDY POPULATION: We conducted an environmental scan in three overdose hotspots’‘ two in an urban center and one in a small city’‘ identified by the Rhode Island Department of Health as having the highest opioid overdose burden in Rhode Island. We engaged hotspot community stakeholders to identify neighborhood factors to map within each hotspot. Locations of addiction treatment, public transportation, harm reduction programs, public facilities , first responders, and social services agencies were converted to latitude and longitude and mapped in ArcGIS. Using Esri Service Areas, we will evaluate the service areas of stationary services. We will overlay overdose events and use logistic regression identify neighborhood factors associated with overdose by comparing hotspot and non-hotspot neighborhoods. RESULTS/ANTICIPATED RESULTS: We anticipate that there will be differing neighborhood characteristics associated with overdose events in the densely populated urban area and those in the smaller city. The urban area hotspots will have overlapping social services, addiction treatment, and transportation service areas, while the small city will have fewer community resources without overlapping service areas and reduced public transportation access. We anticipate that overdoses will occur during times of the day when services are not available. Overall, overdose hotspots will be associated with increased census block level unemployment, homelessness, vacant housing, and low food security. DISCUSSION/SIGNIFICANCE OF FINDINGS: Identifying factors associated with opioid overdoses will enable better resource allocation in communities most impacted by the overdose epidemic. Study results will be used for novel intervention design to prevent opioid overdose deaths in communities with high burden of opioid overdose.