Sham Subjects Research Articles

Global Cerebral Ischemia Induces Short-Term Changes to Estrous Cycle Regularity and Alters Hypothalamic-Pituitary-Gonadal Axis Function

Global cerebral ischemia (GCI), which mimics cardiac arrest in humans, induces numerous functional alterations throughout the brain, but its impact on the hypothalamic-pituitary-gonadal (HPG) axis remains largely unknown. Importantly, the HPG axis modulates sex hormones and neuropeptides, such as estrogen and kisspeptin, as well as estrous cycle regularity. The goals of this study were to determine if estrous cycle regularity is affected by GCI and to characterize HPG axis functionality post-ischemia. Adult female Wistar rats underwent vaginal lavage for 2 weeks to confirm estrous cycle regularity before being subjected to GCI for 10 minutes or sham surgery. Estrous cycle regularity was monitored for 4 weeks following surgery, after which rats were euthanized. Immunofluorescence was used to detect kisspeptin receptors (Kiss1) and estrogen receptors alpha (ERα) and beta (ERβ) expression in the arcuate nucleus (ARC) of the hypothalamus, as well as the CA1 and dentate gyrus (DG) of the hippocampus. Following surgery, all sham subjects maintained a regular estrous cycle, whereas two thirds of ischemic rats presented a disrupted cycle following GCI (persistent diestrus, average length 13 days), before resuming regular cycling. T-test revealed a significant increase in Kiss1 receptors in the ARC of GCI rodents, as well as a trend for higher ERα expression in the CA1 of ischemic rats. Overall, our results highlight that while estrous cycle disruptions resolve within 2 weeks after the ischemic event, HPG axis dysregulation persists up to one month after GCI, hinting at possible long-term impacts on the reproductive axis.

(488) Pooled Outcomes from over 300 Controlled Subjects Reveal Improvements in Sexual Function and a Majority Shift to Lower IPSS Symptom Severity after Treatment with PUL

Abstract Introduction The Prostatic Urethral Lift (PUL) is the most thoroughly studied minimally invasive surgical therapy (MIST) for BPH with over 300 patients studied in a controlled trial setting. Objective In this combined analysis from 5 PUL controlled studies we examine the meta-effect of the procedure on outcomes including sexual function and the proportion of subjects who move from higher to lower IPSS symptom severity grades. Methods Five PUL controlled studies were utilized for the analysis, including L.I.F.T. (RCT: PUL vs Sham [n = 140]), Crossover (crossover of L.I.F.T. sham subjects [n = 51]), MedLift (single arm, subjects with OML [n = 45]), BPH6 (RCT; PUL vs TURP [n=44]), and LOCAL (Single arm, PUL under local anesthesia in office [n=51]]. Sexual function was evaluated with SHIM and MSHQ-EjD, as well as IPSS, QoL, Qmax at baseline and through 12M post-PUL. Post-op AEs, catheterization, surgical retreatment, BPH medication usage, IPSS severity level shift, and IPSS responder rate (≥3pt improvement from baseline) were examined through 12M. Results 331 subjects were treated with PUL among the five studies and had largely similar baseline characteristics. 266 subjects were sexually active at baseline, and notable improvements were found in SHIM (13%, p<0.01) and MSHQ-EjD (30%, p<0.01) at 12mo. Post-PUL, mean IPSS improved at 1, 3, 6, and 12M at least 11.1 points from baseline (48.5%, p <0.0001). QoL improved up to 52.8% (p <0.0001) and Qmax was increased by at least 4.3 ml/sec (p <0.0001). Most AEs were mild to moderate and resolved within 2 weeks. 33% (73/121) of void trial-tested subjects required catheterization (mean duration 3.7d). 15 subjects underwent surgical retreatment and 7 needed BPH medication through 12M post-PUL. 79% (261/331) of subjects were IPSS responders and 66% (219/331) moved into a lower IPSS severity grade (Table 1). Subjects who underwent a shift in symptom severity on average had lower baseline QoL (1.8 vs 3.1, p<0.000), higher Qmax (13.3 vs 11.9, p<0.05) and were treated with more implants (median 5 vs 4 implants, p<0.01). Conclusions The largest analysis of PUL controlled subjects including over 300 men demonstrates the procedure provides significant improvement in IPSS symptom severity grades. Subjects who were sexually active experienced improvements in sexual function through 12 months. Disclosure Yes, this is sponsored by industry/sponsor: Neotract, Inc./Teleflex. Clarification Industry initiated, executed and funded study. Any of the authors act as a consultant, employee or shareholder of an industry for: Neotract, Inc./Teleflex.

(129) Interim Analysis of a Sham-Controlled Randomized, Prospective Study Using Low Intensity Shockwave Therapy (LiSWT) for Improvement of Erectile Function

Abstract Introduction The Urogold 100 MTS™ electrohydraulic shockwave device is FDA cleared for improved blood flow and connective tissue activation; retrospective studies in ED patients have shown positive outcome. Objective To perform an interim analysis of sham and active treatment changes in primary outcome measures of grayscale ultrasound and DUS. Methods A single-blind, sham-controlled, randomized, prospective study in men with ED naïve to acoustic wave therapy was performed. Those meeting inclusion/exclusion were randomized to one of two treatment arms and assigned to active or sham, 2:1 within each arm. Arm 1 consisted of three treatments of 5000 shocks every three weeks, 4 Hz, 0.12 mJ/mm2; arm 2 consisted of 5000, 3000 and 3000 shocks, 4 Hz, 0.12 mJ/mm2; weeks one, two and three respectively, three weeks without treatment, then repeat treatments every three weeks. First follow-up was 20 weeks after initial treatment; DUS and grayscale imaging using a 15.4 MHz probe were repeated under pharmacologic erection 3-4/4 hardness. Post-treatment grayscale percent hypoechoic regions within the corpora cavernosa were assessed: none (0), mild (1), moderate (2) and severe (3) and compared to baseline. Post-treatment EDV and PSV were compared to baseline. Upon completion subjects were unblinded. Subjects assigned to sham were crossed over to the opposite arm for active treatment. Subjects initially in active treatment underwent a second follow-up 32 weeks after initial treatment. Data from each treatment arm were analyzed by two-way repeated measures ANOVA with Geisser-Greenhouse correction. Follow-up pairwise comparisons to baseline were performed using Dunnett’s multiple comparison test. In subjects with one on-treatment assessment, missing data due to early discontinuation from the study were imputed by the “last observation carried forward” method. Results Powered for 60 subjects, recruitment was stopped due to COVID after randomizing 36 subjects (22 active, 14 sham). The proximal penis exhibited greatest improvement (decreased heterogeneity score) on grayscale. The number of subjects with improved erectile tissue grayscale ratings in the proximal region was consistently higher in active treatment versus sham groups (Arm 1 = 88.9% vs. 11.1%; Arm 2 = 40.0% vs. 20.0%, respectively). Sham subjects rolled over to active LiSWT also had improved grayscale ratings (Arm 1 = 33.3% vs. 11.1%; Arm 2 = 40.0% vs. 20.0%). Change in heterogeneity was statistically significant for the proximal region in active treatment Arm 1 at both Week 20 (p=0.005) and Week 32 (p=0.001). (Fig 1) Mean IIEF-EF scores were nominally higher in subjects in active treatment with improved grayscale ratings versus those with no improvement on grayscale. Concerning penile blood flow, improvement after LiSWT greater numbers of patients had higher PSV or lower EDV relative to baseline; greater numbers of patients had no worsening in blood flow parameters. Decrease in EDV reached statistical significance in active treatment Arm 2 at Week 32 (p=0.003). Adverse events were transient. Conclusions Flaccid penile LiSWT appears to be safe and efficacious for treating ED based on statistically significant changes between sham and active treatments in primary outcome measures grayscale ultrasound and DUS. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: TRT, Cynosure.

Procedure-Related Complications in Sham Surgeries for Parkinson's Clinical Trials: A Meta-analysis.

Double-blind, sham-controlled neurosurgical trials for neurodegenerative disorders are debated as an ethical dilemma, particularly regarding subjects randomized to the sham surgery group with general anesthesia. The objective of this study was to examine the safety of sham surgeries in Parkinson's disease (PD) clinical trials through complications related to the procedure. A systematic review and meta-analysis were performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Rates and odds ratios (OR) were compared using random effects analysis. Seven studies, all randomized, double-blind, sham surgery-controlled trials, with 309 patients with PD, were qualitatively and quantitatively analyzed: 141 patients in sham groups and 168 patients in the experimental arms of gene or cell therapy trials. Sham subjects had lower rates of gastrointestinal, positioning, incision-site, respiratory (hypoxic or hypercapnic respiratory failure), cardiovascular, thromboembolism, postoperative cognitive decline, skull fracture, and intracranial or spinal complications when compared with active treatment subjects. Sham subjects, however, had a higher rate of perioperative respiratory infections, such as pneumonia or sinusitis. Further, sham subjects were less likely to experience postoperative cognitive decline (OR, 0.23; 95% confidence interval [CI]: 0.11-0.47), intracranial or spinal complications (OR, 0.10; 95% CI: 0.01-0.75), total major morbidity (OR, 0.30; 95% CI: 0.19-0.47), or overall complications (OR, 0.59; 95% CI: 0.47-0.75) when compared with patients receiving experimental therapy. Patients with PD in the sham surgery control arm of cell transplantation or gene therapy clinical trials have a low risk of procedure-related adverse events overall and fewer complications than patients in the experimental groups. There were no reported deaths attributed to sham surgery-controlled PD clinical trials. © 2023 International Parkinson and Movement Disorder Society.

MP79-11 A SHAM-CONTROLLED RANDOMIZED TRIAL OF LOW INTENSITY SHOCKWAVE THERAPY FOR ERECTILE DYSFUNCTION

You have accessJournal of UrologyCME1 Apr 2023MP79-11 A SHAM-CONTROLLED RANDOMIZED TRIAL OF LOW INTENSITY SHOCKWAVE THERAPY FOR ERECTILE DYSFUNCTION Irwin Goldstein, Sue W. Goldstein, and Noel N. Kim Irwin GoldsteinIrwin Goldstein More articles by this author , Sue W. GoldsteinSue W. Goldstein More articles by this author , and Noel N. KimNoel N. Kim More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003356.11AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: We performed a sham-controlled, randomized prospective trial in men with ED using an electrohydraulic shockwave device FDA cleared for connective tissue activation and improved blood flow. METHODS: This single-blind study was performed in men with ED naïve to acoustic wave and shockwave therapy. Patients were randomized to treatment and assigned to active low intensity shockwave therapy (LiSWT) (4 Hz, 0.12 mJ/mm2) or sham treatment, 2:1. Arm 1 consisted of 3 treatments of 5000 shocks every 3 weeks. Arm 2 consisted of 5000, 3000, and 3000 shocks during weeks 1, 2, and 3, respectively, followed by an identical cycle of treatment 3 weeks later. Doppler ultrasound and grayscale imaging with a 15.4 MHz probe were performed under pharmacologic erection at weeks 20 and 32. Subjects completing sham treatment were unblinded and crossed over to the opposite arm for active treatment. Post-treatment end diastolic velocity (EDV) and peak systolic velocity (PSV) were measured, and visual grading scores were used to assess extent of hypoechoic regions in the corpora cavernosa. Data were analyzed by 2-way repeated measures ANOVA with Geisser-Greenhouse correction. Pairwise comparisons were performed to baseline used Dunnett’s multiple comparison test. Missing data were imputed by “last observation carried forward”. RESULTS: Although powered for 60, recruitment was limited due to COVID and 36 subjects (22 active, 14 sham) were randomized. Sham treatments showed no significant changes in outcome measures. The number of subjects with improved visual grading scores in the proximal region was consistently higher in active LiSWT vs sham (Arm 1=88.9% vs. 11.1%; Arm 2=40.0% vs. 20.0%, respectively) with statistical significance in Arm 1 at weeks 20 (p=0.005) and 32 (p=0.001). Sham subjects rolled over to active LiSWT also had improved grayscale ratings (Arm 1=33.3% vs. 11.1%; Arm 2=40.0% vs. 20.0%). After LiSWT, greater numbers of patients had higher PSV, lower EDV, or no worsening of blood flow parameters relative to baseline. Decrease in EDV was statistically significant in active treatment Arm 2 at Week 32 (p=0.003). Mean IIEF-EF scores were nominally higher in subjects in active treatment who had improved visual grading scores vs those with no improvement. Adverse events were transient. CONCLUSIONS: Flaccid penile LiSWT appears to be safe and efficacious for treating ED based on statistically significant changes from baseline between sham and active treatments in primary outcome measures. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e1145 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Irwin Goldstein More articles by this author Sue W. Goldstein More articles by this author Noel N. Kim More articles by this author Expand All Advertisement PDF downloadLoading ...

High-Fat Diet Exacerbates Stress Responsivity in Juvenile Traumatic Brain Injury.

Abstract Traumatic brain injury (TBI) is one of the leading causes of death for children in the United States. Juveniles are more likely to sustain TBIs than most other age groups, and TBI has been shown to result in increased anxiety and stress behaviors. In addition, the hypothalamic-pituitary-adrenal (HPA) axis has previously been shown to become dysregulated after a TBI. Further, many children consume diets high in saturated fats and refined sugars, which are also connected to alterations in HPA axis function and behavior disorders. The goal of the current study was to identify a potential relationship between high-fat diet (HFD) consumption and TBI on HPA axis function in juvenile rats. In the present study, male juvenile Long-Evans rats were fed either a combination of an HFD with a high-fructose corn syrup solution or a standard chow diet. On post-natal Day 30, subjects sustained either a sham TBI or a TBI via the Closed-Head Injury Model of Engineered Rotational Acceleration (CHIMERA). Subjects participated in a trial of the open field test (OFT) following injury. In addition, some rats performed in an acute restraint stress test. All subjects were euthanized 7 days post-injury. Brain and blood plasma samples were collected for use in real-time polymerase chain reaction (RT-PCR), immunohistochemistry, and corticosterone or adrenocorticotropic hormone (ACTH) assays. Immediately following TBI, injured juveniles had increased time to righting and walking, with HFD-fed TBI rats having increased time to walking over Chow-fed TBI rats. HFD-fed TBI rats had a reduced number of entries to the center of the OFT, in addition to reduced time spent in the center compared with HFD Sham controls and Chow TBI rats. During the acute restraint stress test, HFD-fed TBI rats had elevated pre-stress ACTH and corticosterone and post-stress ACTH levels. Pre-stress ACTH levels were significantly elevated in HFD TBI compared with Chow TBI. Further, pre-stress ACTH:corticosterone ratios were elevated in HFD TBI compared with Chow TBI. cFos immunoreactivity in the paraventricular nucleus (PVN) of the hypothalamus following the acute restraint stress test was elevated in HFD-fed TBI rats. HFD TBI rats had greater activation of cFos in the PVN compared with Chow TBI. In addition, RT-PCR showed significantly reduced expression of relevant HPA axis genes, NR3C1, NR3C2, and CRHR2, in the hypothalamus of TBI subjects compared with Sham subjects. Further, AVP and CRHR2 in the hypothalamus were significantly reduced in HFD TBI compared with Chow TBI. These results offer evidence that TBI paired with high-fat diet consumption can cause HPA axis dysfunction, resulting in more anxiety-like behaviors.

P-TS005. Potential of transcranial direct current stimulation in modulating spinal reflex excitability in persons with chronic stroke

Introduction. Stroke is the main cause of disability in adults living in developing countries. Spasticity is a primary impairment of stroke with an effect on the activities of daily life. Neural changes of the paretic limb often occur following spasticity. The aim of this study was to investigate the effect of Transcranial Direct Current Stimulation on neural properties of the spastic muscle in patients with chronic stroke. Methods. In this study double-blinded trial 22 participants with >6 months after stroke were randomized into two groups: active anodal or sham tDCS. The subjects in the experimental group participated in active tDCS and balance training program of 5 sessions. Evaluation performed at baseline and at the end of the intervention. Hmax/Mmax ratio and H-reflex latency were assumed. Clinical assessment was conducted using the Modified Ashworth Scale (MAS), Timed Up and Go (TUG), as well as the Timed 10-Meter Walk Test (10MWT). Repeated measure analysis of variance was used in the analysis. Friedman test in SPSS version 22.0 was used to analysis the response of ordinal variable in different stages of evaluation. Results. The active tDCS group showed significant differences in the Hmax/Mmax (P<0.005), 10MWT (P<0.01) Scale from sham subjects. Also, the latency of the H response from the stimulation was increased for the experimental patients compared to sham. Time by group interaction in the 10MWT, Hmax/Mmax ratio and H-reflex latency was significant. Nevertheless, there were no significant differences in the TUG (P=0.30) scores. The results indicated that the H-reflex latency, Hmax/Mmax ratio were altered after a-tDCS intervention which is correlated with improvement in walking outcome. Conclusion. The findings of the study illustrate that tDCS treatment with balance training can improve reflexes, and walking capacity in chronic stroke and can be considered as a promising therapeutic tool in rehabilitation setting.

Effect of adipose graft transposition procedure (AGTP) on the ischemic arrhythmogenic substrate: an MRI study in a swine model of chronic myocardial infraction

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Instituto de Salud Carlos III BACKGROUND Cardiac regenerative therapy is a promising treatment for patients with myocardial infarction (MI) and heart failure. Nevertheless, previous ex-vivo studies have raised concern on the potential increased risk of arrhythmic events following certain cell therapies. Adipose graft transposition procedure (AGTP) is a cardiac reparative therapy consisting in transposing a vascularized adipose flap from the autologous pericardium and placing it over the epicardial scar area and has demonstrated to reduce infarct size and improve the left ventricular ejection fraction in preclinical and human studies. PURPOSE To assess the effect of the AGTP on the post-MI scar composition and image-based ventricular tachycardia (VT) corridors detection by means of late gadolinium enhanced cardiac magnetic resonance (LGE-CMR). METHODS A left circumflex artery (first marginal branch) MI was induced in 9 Landrace X Large White Pigs by delivering 1-3 coils. Two weeks post-MI, all subjects underwent a 3 Tesla LGE-CMR and randomized to the AGTP or sham group. LGE-CMR was repeated 30 days post-treatment (6 weeks post-MI). The arrhythmogenic substrate was characterized with an advanced image post-processing tool (ADAS 3D) and included quantification of dense scar and border zone (BZ) mass and detection of ventricular tachycardia (VT) corridors (including corridor scar mass). RESULTS The overall scar mass did not differ between scans in the overall population (7.6 ± 3.5 g vs 7.5 ± 2.2 g in the baseline and post-treatment scans, respectively; p = 0.9). Compared to the sham subjects, those receiving AGTP showed an absolute reduction of the total (-3.2 ± 1.4 g vs. +2.4 ± 1.7 g, p = 0.04) and dense scar (-0.9 ± 0.4 g vs. +0.7 ± 0.5 g, p = 0.03). BZ mass tended to decrease in the AGTP group (-2.2 vs 1.63 g; p = 0.06). The AGTP group showed a trend to reduce the number of VT corridors (-1 ± 0.7 vs. +0.4 ± 0.2, p = 0.078) and corridor scar mass (-0.3 ± 0.26 g vs. +0.1 ± 0.1 g, p = 0.11) (figure). CONCLUSIONS Cardiac reparative therapy of MI with AGTP reduced dense scar mass, compared to the increase observed in the sham group. The trend to reduce the BZ mass and the number/mass of VT corridors suggests a beneficial effect on the arrhythmic remodeling of the post-MI scar. Abstract Figure. Reduction in corridor"s number

Passive Heat Therapy Reduces Aortic and Carotid Artery Stiffness and Intima‐Media Thickness in Middle‐Aged and Older Adults

Aging is accompanied by stiffening of the large elastic arteries and adverse arterial remodeling, which increase risk for cardiovascular diseases (CVD). Passive heat therapy—chronic use of hot baths and saunas—is associated with lower CVD risk. However, it is unknown if a heat therapy intervention affects arterial stiffness and structure in middle-aged and older (MA/O) adults without overt clinical disorders. Purpose To determine if passive heat therapy, via hot water immersion, in healthy MA/O adults: 1) reduces arterial stiffness, as measured by carotid-femoral pulse wave velocity (c-f PWV; i.e., aortic stiffness) and common carotid artery (CCA) and superficial femoral artery (SFA) β-stiffness index; and 2) alters arterial structure, as measured by CCA and SFA intima-media thickness (IMT) and vessel diameter. Methods 23 healthy, minimally active MA/O men and postmenopausal women (57-79 years) were randomized to participate in 30, 60-minute sessions (3-4x/week over 8-10 weeks) of either hot water immersion (40.5°C, N=12, 5M/7F, 68±2 years) or thermoneutral water immersion (36°C, “sham”, N=11, 4M/7F, 66±2 years). Ultrasound- and tonometry-based measures of arterial stiffness and structure were measured at baseline and 24-72 hours after the final water immersion session. Data are mean ± SEM; P-values were determined by 2-way mixed-design ANOVA with Tukey's post-hoc tests. Results In the large elastic arteries (i.e., the arteries that stiffen with aging), heat therapy reduced c-f PWV from 8.7±0.6 to 8.0±0.7 m/s (p=0.03) and CCA β-stiffness index from 10.6±1.6 to 7.8±0.8 (p=0.02); however, heat therapy did not change SFA β-stiffness index (15.0±1.6 vs. 14.6±1.6, p=0.99), a more muscular artery. Heat therapy also improved selective measures of arterial structure. In the CCA, heat therapy decreased IMT from 0.70±0.01 to 0.66±0.02 mm (p<0.01) without affecting diameter (7.4±0.2 vs. 7.2±0.2 mm, p=0.90). Conversely, in the SFA, heat therapy did not alter IMT (0.56±0.02 vs. 0.57±0.02 mm, p>0.99), but tended to increase diameter from 7.6±0.2 to 7.9±0.3 mm (p=0.06), which likely occurred due to repeated increases in vascular shear stress to the lower body during hot water immersion sessions. No changes were observed in any outcome in sham subjects (c-f PWV: 8.5±0.5 vs. 8.4±0.4 m/s, p=0.85; CCA β-stiffness index: 10.0±2.2 vs. 9.9±2.6, p=0.96; CCA IMT: 0.70±0.02 vs. 0.70±0.02 mm, p=0.92; SFA β-stiffness index: 15.2±2.1 vs. 16.0±2.9, p=0.93; CCA diameter: 7.5±0.1 vs. 7.4±0.1 mm, p=0.42; SFA IMT: 0.57±0.03 vs. 0.56 ±0.03 mm, p=0.85; SFA diameter: 7.8±0.4 vs. 7.9±0.3 mm, p=0.98). Conclusion Heat therapy subjects demonstrated clinically relevant reductions in aortic and carotid artery stiffness, and in CCA IMT, all of which are independent predictors of future CVD risk. As such, heat therapy may be a promising lifestyle-based intervention for improving arterial health and thereby reducing CVD risk in MA/O adults.

Optogenetically transduced human ES cell-derived neural progenitors and their neuronal progenies: Phenotypic characterization and responses to optical stimulation.

Optogenetically engineered human neural progenitors (hNPs) are viewed as promising tools in regenerative neuroscience because they allow the testing of the ability of hNPs to integrate within nervous system of an appropriate host not only structurally, but also functionally based on the responses of their differentiated progenies to light. Here, we transduced H9 embryonic stem cell-derived hNPs with a lentivirus harboring human channelrhodopsin (hChR2) and differentiated them into a forebrain lineage. We extensively characterized the fate and optogenetic functionality of hChR2-hNPs in vitro with electrophysiology and immunocytochemistry. We also explored whether the in vivo phenotype of ChR2-hNPs conforms to in vitro observations by grafting them into the frontal neocortex of rodents and analyzing their survival and neuronal differentiation. Human ChR2-hNPs acquired neuronal phenotypes (TUJ1, MAP2, SMI-312, and synapsin 1 immunoreactivity) in vitro after an average of 70 days of coculturing with CD1 astrocytes and progressively displayed both inhibitory and excitatory neurotransmitter signatures by immunocytochemistry and whole-cell patch clamp recording. Three months after transplantation into motor cortex of naïve or injured mice, 60–70% of hChR2-hNPs at the transplantation site expressed TUJ1 and had neuronal cytologies, whereas 60% of cells also expressed ChR2. Transplant-derived neurons extended axons through major commissural and descending tracts and issued synaptophysin+ terminals in the claustrum, endopiriform area, and corresponding insular and piriform cortices. There was no apparent difference in engraftment, differentiation, or connectivity patterns between injured and sham subjects. Same trends were observed in a second rodent host, i.e. rat, where we employed longer survival times and found that the majority of grafted hChR2-hNPs differentiated into GABAergic neurons that established dense terminal fields and innervated mostly dendritic profiles in host cortical neurons. In physiological experiments, human ChR2+ neurons in culture generated spontaneous action potentials (APs) 100–170 days into differentiation and their firing activity was consistently driven by optical stimulation. Stimulation generated glutamatergic and GABAergic postsynaptic activity in neighboring ChR2- cells, evidence that hChR2-hNP-derived neurons had established functional synaptic connections with other neurons in culture. Light stimulation of hChR2-hNP transplants in vivo generated complicated results, in part because of the variable response of the transplants themselves. Our findings show that we can successfully derive hNPs with optogenetic properties that are fully transferrable to their differentiated neuronal progenies. We also show that these progenies have substantial neurotransmitter plasticity in vitro, whereas in vivo they mostly differentiate into inhibitory GABAergic neurons. Furthermore, neurons derived from hNPs have the capacity of establishing functional synapses with postsynaptic neurons in vitro, but this outcome is technically challenging to explore in vivo. We propose that optogenetically endowed hNPs hold great promise as tools to explore de novo circuit formation in the brain and, in the future, perhaps launch a new generation of neuromodulatory therapies.

Brain Death Enhances Activation of the Innate Immune System and Leads to Reduced Renal Metabolic Gene Expression.

Brain death (BD)-associated inflammation has been implicated in decreased kidney allograft function and survival, but the underlying mechanisms have not been well distinguished from the conditions of critical care itself. We have developed a clinically translatable model to separate and investigate strategies to improve donor management and critical care. Brain-dead (n = 12) and sham (n = 5) rhesus macaques were maintained for 20 hours under intensive care unit-level conditions. Samples were collected for immunophenotyping, analysis of plasma proteins, coagulation studies, and gene analysis for changes in immune and metabolic profile with comparison to naive samples (n = 10). We observed an increase in circulating leukocytes and cytokines, activation of complement and coagulation pathways, and upregulation of genes associated with inflammation in both brain-dead and sham subjects relative to naïve controls. Sham demonstrated an intermediate phenotype of inflammation compared to BD. Analysis of gene expression in kidneys from BD kidneys revealed a similar upregulation of inflammatory profile in both BD and sham subjects, but BD presented a distinct reduction in metabolic and respiratory processes compared to sham and naïve kidneys. BD is associated with activation of specific pathways of the innate immune system and changes to metabolic gene expression in renal tissue itself; however, sham donors presented an intermediate inflammatory response attributable to the critical care environment. The early onset and penetrating impact of this inflammatory response underscores the need for early intervention to prevent perioperative tissue injury to transplantable organs.

Passive Heat Therapy Lowers Systolic Blood Pressure and Improves Vascular Endothelial Function in Healthy Older Adults

Aging is associated with elevated systolic blood pressure (SBP) and impaired vascular function, characterized by endothelial dysfunction and stiffening of the large elastic arteries, which all increase risk for cardiovascular diseases (CVD). Passive repeated heat exposure (“heat therapy”) is associated with lower CVD risk at the population level and improves CV function in animals, healthy young adults, and selective groups of patients with clinical disease. It is unknown, however, if heat therapy lowers SBP and/or improves vascular function in late middle‐aged to older (MA/O) adults free of clinical diseases/disorders.PurposeTo determine if heat therapy via hot water immersion in healthy MA/O adults lowers SBP and/or improves markers of vascular health that are independent risk factors for CVD.MethodsTen healthy MA/O men and post‐menopausal women (57–76 years) were randomly assigned to participate in thirty 60‐min sessions of either hot (40.5°C; N=6, 3M/3F, 67±3 years) or thermoneutral (36°C, “sham”; N=4, 1M/3F, 69±2 years) water immersion. Resting (casual) BP and ultrasound‐ and tonometry‐based measures of vascular function and structure were measured at baseline and 24–72 h after the last water immersion session. Data are mean±S.E. Changes in outcome variables were compared within groups by paired t‐test.ResultsHot water immersion increased rectal temperature (Tre) from 36.9±0.2 to 38.7±0.1°C (p&lt;0.001; data from first session), whereas no changes in Tre were observed in sham subjects (resting: 37.2±0.1 vs. peak: 37.3±0.1°C, p=0.23). Adherence was good, with subjects completing all sessions in 9.8±0.4 weeks (3.1±0.1 sessions/week). Heat therapy was well‐tolerated, with only 3% of sessions ending early due to occurrence of mild heat‐related symptoms (e.g., lightheadedness). Heat therapy decreased SBP by 10 mmHg on average (130±4 vs. 120±5 mmHg, p=0.04), increased endothelial function by ~50% (brachial artery flow‐mediated dilation, FMDBA: 4.4±0.7 vs. 6.5±0.9%, p&lt;0.01), and decreased carotid intima media thickness (cIMT) by ~10% (0.70±0.02 vs. 0.64±0.03 mm, p=0.02), but did not significantly alter arterial stiffness (carotid‐femoral pulse wave velocity, PWV: 8.6±0.4 vs. 8.2±0.7 m/s, p=0.48), or diastolic BP (DBP: 81±4 vs. 78±5 mmHg, p=0.36). No effects were observed in sham subjects (SBP: 133±11 vs. 131±12 mmHg, p=0.31; FMDBA: 4.9±0.9 vs. 4.8±0.7%, p=0.92; cIMT: 0.69±0.02 vs. 0.68±0.03 mm, p=0.77; PWV: 9.6±1.6 vs. 9.3±1.6 m/s, p=0.35; DBP: 79±3 vs. 74±2 mmHg, p=0.17).ConclusionOur preliminary results suggest that in healthy MA/O adults, heat therapy is safe, is associated with excellent adherence, and induces clinically‐significant reductions in casual SBP and improvements in endothelial function comparable to or greater than those typically observed with aerobic exercise training. Heat therapy also induces a unique reduction in cIMT typically found only with longer‐term pharmacological therapies. Heat therapy may be a promising lifestyle‐based strategy for reducing SBP, endothelial dysfunction and, possibly, CVD risk with aging.Support or Funding InformationNCT03264508This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Increased Cortical Interleukin‐1 Receptor Signaling Following Blast‐Induced Traumatic Brain Injury

Traumatic Brain Injury (TBI) is a leading cause of disability in the United States resulting in an estimated annual economic cost of over $75 billion. Furthermore, there is a lack of any FDA approved medications aimed at ameliorating the effects of acute injury or the chronic comorbidities of TBI. Those affected are often left with profound chronic neurologic deficits such as major depressive disorder (MDD), anxiety, sleep disturbances, and/or post‐traumatic epilepsy. Neuroinflammation is a hallmark of TBI, although whether neuroinflammatory signaling contributes to the etiology of neuropsychiatric complications following various forms of neurotrauma is not well characterized. We hypothesize that following TBI, interleukin‐1 alpha (IL‐1α) and interleukin‐1 beta (IL‐1β) signaling through functional type 1 interleukin‐1 receptors (IL‐1R1) acts to orchestrate molecular signaling that contributes to the generation of neuropsychiatric sequelae. To test this hypothesis, the current study aimed to characterize alterations in interleukin‐1 (IL‐1) signaling elicited by blast‐induced TBI in murine subjects. Moderate blast‐induced TBI (≈ 20 psi) resulted in an acute increase in righting reflex time (RRT) and a transient decrease in total locomotor activity 3 hrs post‐injury. Rapid increases in il‐1β, il‐1α and tnf‐α mRNA expression occurred in cortical and hippocampal samples ipsilateral to injury in TBI subjects as compared to their sham counterparts, indicating increased CNS production of proinflammatory cytokines. Elevated levels of il‐1r1 mRNA were found in the ipsilateral cortex following injury, however no changes were found in interleukin‐1 receptor associated kinase‐4 (irak‐4) mRNA expression, providing evidence of increased cortical IL‐1R1‐mediated signaling in TBI subjects. Further, TBI resulted in increased mRNA expression and immunoreactivity of the astrocytic marker glial fibrillary acidic protein (GFAP) in the ipsilateral cortex as compared to sham subjects 24 and 72 hrs post‐injury, respectively, indicative of significant reactive astrogliosis. One‐week post‐injury, TBI increased despair‐like behavior as compared to sham treatments as determined by the Porsolt forced swim test (FST). Importantly, no concomitant alterations in general locomotor activity were found in TBI subjects one‐week post‐injury. Future studies will utilize murine models, affording the conditional elimination and restoration of IL‐1R1 in conjunction with blast‐induced TBI to ascertain the cell‐specific contributions of IL‐1R1‐mediated signaling to injury‐induced neuroinflammation and functional outcomes post‐injury. Systematic characterization of IL‐1R1 signaling post‐injury will lead to a better understanding of how neuroinflammatory signaling contributes to the acute and chronic neurologic sequelae of TBI and may lead to viable pharmacotherapeutic targets aimed at ameliorating these deficits.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Peripheral Inflammation Accelerates the Onset of Mechanical Hypersensitivity after Spinal Cord Injury and Engages Tumor Necrosis Factor α Signaling Mechanisms.

Previously, we showed that noxious stimulation of the tail produces numerous detrimental effects after spinal cord injury (SCI), including an earlier onset and increased magnitude of mechanical hypersensitivity. Expanding on these observations, this study sought to determine whether localized peripheral inflammation similarly impacts the expression of mechanical hypersensitivity after SCI. Adult rats received a moderate contusion injury at the thoracic level (Tl0) or sham surgery, and were administered complete Freund's adjuvant (CFA) or vehicle in one hindpaw 24 hours later. Examination of locomotor recovery (Basso, Beattie, and Bresnahan [BBB] score) showed no adverse effect of CFA. Mechanical testing with von Frey hairs was done at time-points ranging from 1 h to 28 days after CFA or vehicle treatment, and rats were sacrificed at 1, 7, or 28 days for cellular assessment. Unlike vehicle-treated SCI rats where mechanical hypersensitivity emerged at 14 days, CFA-treated SCI rats showed mechanical hypersensitivity as early as 1 h after CFA administration, which lasted at least 28 days. CFA-treated sham subjects also showed an early onset of mechanical hypersensitivity, but this was maintained up to 7 days after treatment. Cellular assessments revealed congruent findings. Expression levels of c-fos, tumor necrosis factor α (TNFα), TNF receptors, and members of the TNFα signaling pathway such as caspase 8 and phosphorylated extracellular related kinase (pERK) were preferentially upregulated in the lumbar spinal cord of SCI-CFA rats. Meanwhile, c-jun was significantly increased in both CFA-treated groups. Overall, these results together with our previous reports, suggest that peripheral noxious input after SCI facilitates the development of pain by mechanisms that may require TNFα signaling.

Proteomic Analysis of Urothelium of Rats with Detrusor Overactivity Induced by Bladder Outlet Obstruction

Overactive bladder (OAB) syndrome is a condition that has four symptoms: urgency, urinary frequency, nocturia, and urge incontinence and negatively affects a patient's life. Recently, it is considered that the urinary bladder urothelium is closely linked to pathogenesis of OAB. However, the mechanisms of pathogenesis of OAB at the molecular level remain poorly understood, mainly because of lack of modern molecular analysis. The goal of this study is to identify a potential target protein that could act as a predictive factor for effective diagnosis and aid in the development of therapeutic strategies for the treatment of OAB syndrome. We produced OAB in a rat model and performed the first proteomic analysis on the mucosal layer (urothelium) of the bladders of sham control and OAB rats. The resulting data revealed the differential expression of 355 proteins in the bladder urothelium of OAB rats compared with sham subjects. Signaling pathway analysis revealed that the differentially expressed proteins were mainly involved in the inflammatory response and apoptosis. Our findings suggest a new target for accurate diagnosis of OAB that can provide essential information for the development of drug treatment strategies as well as establish criteria for screening patients in the clinical environment.

Remote Ischemic Preconditioning Does Not Prevent White Matter Injury in Neonates

Remote ischemic preconditioning (RIPC) is a mechanism to protect tissues from injury during ischemia and reperfusion. We investigated the neuroprotective effects of RIPC in neonates undergoing cardiac surgery. The outcome was white matter injury (WMI), assessed by the change in volume of WMI from preoperative to postoperative brain magnetic resonance imaging (MRI). Patients were randomized to RIPC or SHAM. RIPC was induced prior to cardiopulmonary bypass by four 5-minute cycles of blood pressure cuff inflation to produce ischemia of the lower extremity. For patients randomized to SHAM, the cuff was positioned, but not inflated. The study included 67 patients, with 33 randomized to RIPC and 34 randomized to SHAM. Preoperative and postoperative MRIs were available in 50 patients, including 26 of the 33 RIPC patients and 24 of the 34 SHAM patients. There were no differences in baseline and operative characteristics for either the overall study group or the group with evaluable MRIs. WMI was identified in 28% of the patients preoperatively and in 62% postoperatively. There was no difference in the prevalence of WMI by treatment group (p > 0.5). RIPC patients had an average change in WMI of 600 mL3, and SHAM subjects had an average WMI change of 107 mL3. There was no significant difference in the mean value of WMI change between patients who received RIPC and those who received SHAM treatments (p= 0.178). In this randomized, blinded clinical trial, there was no evidence that use of RIPC provides neuroprotection in neonates undergoing repair of congenital heart defects with cardiopulmonary bypass.

Effect of Single-Session, Cryogen-Cooled Monopolar Radiofrequency Therapy on Sexual Function in Women with Vaginal Laxity: The VIVEVE I Trial.

Objective: This subanalysis of the VIVEVE I trial aimed to evaluate the impact of cryogen-cooled monopolar radiofrequency (CMRF) therapy, for the treatment of vaginal laxity, on the domains of sexual function included in the Female Sexual Function Index (FSFI).Materials and Methods: The VIVEVE I clinical trial was prospective, randomized, single-blind, and Sham-controlled. Nine clinical study centers in Canada, Italy, Spain, and Japan were included. This subanalysis included premenopausal women with self-reported vaginal laxity who had ≥1 term vaginal delivery and a baseline FSFI total score ≤26.5, indicating sexual dysfunction. Enrolled subjects were randomized (2:1) to receive CMRF therapy [Active (90 J/cm2) vs. Sham (≤1 J/cm2)] delivered to the vaginal tissue. Independent analyses were conducted for each FSFI domain to evaluate both the mean change, as well as the clinically important change for Active- versus Sham-treated subjects at 6 months post-intervention.Results: Subjects randomized to Active treatment (n = 73) had greater improvement than Sham subjects (n = 35) on all FSFI domains of sexual function at 6 months postintervention. The analysis of covariance change from baseline analyses showed statistically significant improvements, in favor of Active treatment, for sexual arousal (p = 0.004), lubrication (p = 0.04), and orgasm (p = 0.007). In addition, Active treatment was associated with clinically important and statistically significant improvements in sexual desire [Odds ratio (OR) = 3.01 (1.11–8.17)], arousal [OR = 2.73 (1.06–7.04)], and orgasm [OR = 2.58 (1.08–6.18)].Conclusions: This subanalysis showed CMRF therapy is associated with statistically significant and clinically important improvements in sexual function in women with vaginal laxity. These findings provide the first randomized, placebo-controlled energy-based device evidence for functional improvements associated with a nonsurgical modality for a highly prevalent and undertreated condition.

Abstract P395: A New Technique for Selective Denervation of the Aortic and Carotid Baroreceptors

The traditional sinoaortic denervation (SAD) technique in rats includes aortic and carotid baroreceptors denervation, but also leads to concomitant denervation of the carotid chemoreceptors. Here we report a new technique to denervate the carotid and aortic baroreceptors, selectively, preserving the carotid chemoreceptors intact. Wistar rats were subjected to selective aortic and carotid baroreceptor denervation sparing the carotid chemoreceptors intact (BAROS-X), or sham surgery (SHAM). The animals were also implanted with femoral arterial and venous catheter for arterial pressure recording and drug administration. In unanesthetized, freely moving rats, baroreflex or peripheral chemoreflex activation was elicited by intravenous injection of phenylephrine or potassium cyanide (KCN), respectively. Phenylephrine caused a significant hypertensive response in SHAM (Δ = 46 ± 3 mmHg, P&lt;0.001) and BAROS-X (Δ = 38 ± 3 mmHg, P&lt;0.001) combined with a reflex bradycardia in SHAM (Δ = -54 ± 11 bpm, P&lt;0.001) but no change in heart rate in BAROS-X subjects (Δ = -5 ± 2 bpm, ns); confirming the absence of aortic and carotid baroreceptors in BAROS-X rats. KCN elicited significant hypertensive response in SHAM (44 ± 6 mmHg, P&lt;0.001) and BAROS-X (39 ± 5 mmHg, P&lt;0.001) subjects followed by bradycardic response in SHAM (-88 ± 17 bpm, P&lt;0.001) and BAROX-S (-103 ± 23 bpm, P&lt;0.01) rats; confirming that the carotid chemoreceptors were intact in both groups. These data provide support to a new surgical technique that allows working with conscious rats exhibiting selective inactivation of the aortic and carotid baroreceptors, combined with intact carotid chemoreceptors.

Evidence of Altered Trigeminal Nociception in an Animal Model of Fibromyalgia.

Fibromyalgia (FM) is a debilitating chronic condition that significantly affects quality of life. A strong association has been demonstrated between FM and chronic pain in the trigeminal region in clinical studies. This study was performed to evaluate the response to acute and chronic noxious stimuli applied to the facial region. Adult male Wistar rats (250-270 g, N = 10 for each group) were used in the current study. A subchronic swim stress model was used as the animal model of FM. Anxiety-like behaviors and response to acute and chronic noxious stimuli were assayed using the elevated plus maze, eye wiping test, and orofacial formalin test, respectively. Balance and motor function were evaluated using rotarod and wire grip tests. An increased anxiety-like behavior was observed in swim stress rats in comparison with control and sham subjects. Response to acute and chronic noxious stimuli in the trigeminal region was increased in the stressed rats. Motor and balance function were not altered following stress. Results of the current study demonstrated a hyperalgesic state in the trigeminal region in a possible animal model of FM. This study provides a reliable animal model for further research on the possible mechanisms of orofacial pain in FM.

Post-infarct sleep disruption and its relation to cardiac remodeling in a rat model of myocardial infarction

ABSTRACTSleep disruption after myocardial infarction (MI) by affecting ubiquitin–proteasome system (UPS) is thought to contribute to myocardial remodeling and progressive worsening of cardiac function. The aim of current study was to test the hypothesis about the increased risk of developing heart failure due to experience of sleep restriction (SR) after MI. Male Wistar rats (n = 40) were randomly assigned to four experimental groups: (1) Sham, (2) MI, (3) MI and SR (MI + SR) (4) Sham and SR (Sham + SR). MI was induced by permanent ligation of left anterior descending coronary artery. Twenty-four hours after surgery, animals were subjected to chronic SR paradigm. Blood sampling was performed at days 1, 8 and 21 after MI for determination of serum levels of creatine kinase-MB (CK-MB), corticosterone, malondialdehyde (MDA) and nitric oxide (NO). Finally, at 21 days after MI, echocardiographic parameters and expression of MuRF1, MaFBx, A20, eNOS, iNOS and NF-kB in the heart were evaluated. We used H&E staining to detect myocardial hypertrophy. We found out that post infarct SR increased corticosterone levels. Our results highlighted deteriorating effects of post-MI SR on NO production, oxidative stress, and echocardiographic indexes (p < 0.05). Moreover, its detrimental effects on myocardial damage were confirmed by overexpression of MuRF1, MaFBx, iNOS and NF-kB (p < 0.001) in left ventricle and downregulation of A20 and eNOS (p < 0.05). Furthermore, histological examination revealed that experience of SR after MI increased myocardial diameter as compared to Sham subjects (p < 0.05). Our data suggest that SR after MI leads to an enlargement of the heart within 21 days, marked by an increase in oxidative stress and NO production as well as an imbalance in UPS that ultimately results in cardiac dysfunction and heart failure.