AIMS:Underactive bladder and bowel function are common in the elderly. Neurokinin 2 receptor (NK2R) agonists induce voiding in young adult animals; however, these effects have not been sufficiently examined in aged animals. Most NK2R agonists also have activity at neurokinin 1 receptors, which produces off target effects, such as transient hypotension. Hypotension is problematic in the elderly due to increased injury risk when falling. Therefore, the aims of this study were to (1) test the ability of NK2R agonists to increase bladder and colorectal pressure without affecting blood pressure and (2) determine if daily administration of a NK2R agonist consistently induces urination and defecation in aged rats. METHODS:Voiding cystometry and manometry were used to evaluate effects of [Arg5, MeLeu9, Nle10]-NKA(4-10) (AMN-NKA) and GR64349 on bladder and colorectal physiology, respectively, while recording blood pressure in anesthetized adult and aged F344 rats. The rapid voiding detection assay was used to test the efficacy of repeated intramuscular GR64349 dosing to induce urination and defecation in aged rats. RESULTS:Intravenous AMN-NKA or GR64349 both increased bladder and colorectal pressure in anesthetized young adult and aged rats, but AMN-NKA produced transient hypotension. Intramuscular GR64349 produced rapid increases in bladder and colorectal pressures in aged and adult rats (10-300 µg/kg) and consistently induced defecation and urination in awake aged rats across a 2 week period of dosing (100 μg/kg/2 times/day). CONCLUSIONS:The results show that GR64349 could be a promising therapeutic for inducing urination and defecation in the elderly without side effects such as hypotension.

One cannot survive without regularly urinating and defecating. People with neurologic injury (spinal cord injury, traumatic brain injury, stroke) or disease (multiple sclerosis, Parkinson's disease, spina bifida) and many elderly are unable to voluntarily initiate voiding. The great majority of them require bladder catheters to void urine and "manual bowel programs" with digital rectal stimulation and manual extraction to void stool. Catheter-associated urinary tract infections frequently require hospitalization, whereas manual bowel programs are time consuming (1 to 2 hours) and stigmatizing and cause rectal pain and discomfort. Laxatives and enemas produce defecation, but onset and duration are unpredictable, prolonged, and difficult to control, which can produce involuntary defecation and fecal incontinence. Patients with spinal cord injury (SCI) consider recovery of bladder and bowel function a higher priority than recovery of walking. Bladder and bowel dysfunction are a top reason for institutionalization of elderly. Surveys indicate that convenience, rapid onset and short duration, reliability and predictability, and efficient voiding are priorities of SCI individuals. Despite the severe, unmet medical need, there is no literature regarding on- demand, rapid-onset, short-duration, drug-induced voiding therapies. This article provides in-depth discussion of recent discovery and development of two candidates for on-demand voiding therapies. The first, [Lys3,Gly8,-R-c-lactam-Leu9]-NKA(3-10) (DTI-117), a neurokinin2 receptor agonist, induces both urination and defecation after systemic administration. The second, capsaicin (DTI-301), is a transient receptor potential cation channel subfamily V member 1 (TRPV1) receptor agonist that induces defecation after intrarectal administration. The review also presents clinical studies of a combination drug therapy administered via iontophoresis and preclinical studies of neuromodulation devices that induce urination and defecation.

Substantial clinical and preclinical evidence indicates that transient receptor potential vanilloid 1 (TRPV1) receptors are expressed on terminals of colorectal chemoreceptors and mechanoreceptors and are involved in various rectal hypersensitivity disorders with common features of colorectal overactivity. These stimulatory properties of TRPV1 receptors on colorectal function suggested that brief stimulation of TRPV1 might provide a means of pharmacologically activating the colorectum to induce defecation in patients with an "unresponsive" colorectum. The current studies explored the basic features of TRPV1 receptor-induced contractions of the colorectum in anesthetized rats with and without acute spinal cord injury (aSCI). Cumulative concentration-response curves to intrarectal (IR) capsaicin (CAP) solutions (0.003%-3.0%) were performed in anesthetized aSCI and spinal intact rats. CAP produced an "inverted U," cumulative concentration-response curve with a threshold for inducing colorectal contractions at 0.01% and a peak response at 0.1% and slight decreases in responses up to 3%. Decreases in responses with concentrations >0.1% are due to a rapid desensitization (i.e., <= 30 minutes) of TRPV1 receptors to each successive dose. Desensitization appeared fully recovered within 24 hours in spinal intact rats. Colorectal contractions were completely blocked by atropine, indicating a reflexogenic activation of parasympathetic neurons, and responses were completely unaffected by a neurokinin 2 receptor antagonist, indicating that release of neurokinin A from afferent terminals and subsequent direct contractions of the smooth muscle was not involved. IR administration of three other TRPV1 receptor agonists produced similar results as CAP.

The feasibility of eliciting defecation and urination after intranasal (IN) or sublingual (SL) delivery of a small peptide NK2 receptor agonist, [Lys5, MeLeu9, Nle10]-NKA(4-10), was examined using prototype formulations in dogs. In anesthetized ani-mals, administration of 100 or 300 a.g/kg IN or 2.0-6.7 mg/kg SL increased colorectal peak pressure and area under the curve. Peak bladder pressure was also increased at the same doses, and this was accompanied by highly efficient voiding at normal physiological bladder pressure. The onset of these effects was rapid (<2.5 min), and the primary contractions lasted similar to 25 min, returning to baseline in <60 min. Slight hypotension lasting a few minutes and causing <10% change from baseline was de-tected after higher doses and was statistically significant after only 100 a.g/kg IN. In conscious dogs, there was a dose-related increase in voiding responses and reduction in the latency to urinate and defecate after 300 and 1000 a.g/kg IN; emesis was also observed at these doses. SL administration of 6.7 mg/kg induced urination within 10 min, but not defecation or emesis. These findings support the feasibility of developing a convenient dosage form of small peptide NK2 receptor agonists as on-demand defecation or urination therapies.

Study design Preclinical pharmacology. Objectives To determine whether blocking substance P signaling attenuates the hypertension and bradycardia evoked by colorectal distension (CRD) in spinal cord injured (SCI) rats. Setting University laboratory in Pennsylvania, U.S.A. Methods Tachykinin NK1 receptor antagonists were administered 30 min prior to CRD three weeks after complete spinal cord transection at the 4 th thoracic (T4) level. The dose range, route of administration, and pretreatment time was based on published data demonstrating occupancy of brain NK1 receptors in rodents. Results Subcutaneous (SC) administration of 10–30 mg/kg GR205171 ((2 S ,3 S )- N -[[2-methoxy-5-[5-(trifluoromethyl)tetrazol-1-yl]phenyl]methyl]-2-phenylpiperidin-3-amine dihydrochloride) reduced CRD-induced hypertension and bradycardia by 55 and 49%, respectively, compared with pretreatment values. There was no effect of GR205171 on resting blood pressure or heart rate. In contrast, the same dose range of CP-99,994 ((2 S ,3 S )- N -[(2-methoxyphenyl)methyl]-2-phenyl-3-piperidinamine dihydrochloride) had no effect on CRD-induced cardiovascular responses. Conclusions The effective dose range of GR205171 to alleviate autonomic dysreflexia is consistent with the blockade of NK1 receptors on pelvic sensory afferents in the lumbosacral spinal cord, which may in turn prevent the over-excitation of sympathetic preganglionic neurons (SPNs) that regulate blood pressure and heart rate. The findings provide preclinical support for the utility of NK1 receptor antagonists to treat autonomic dysreflexia in people with SCI. The difference in the effects of the two NK1 receptor antagonists may reflect the ~200-fold lower affinity of CP-99,994 than GR205171 for the rat NK1 receptor.

The European Society of Organ Transplantation (ESOT) strives to promote equity, diversity, and inclusion (EDI) across all its activities. We surveyed the transplant community's experiences and perspectives regarding EDI within ESOT as an organization and its educational activities, and research in general. A total of 299 respondents completed the questionnaire. About half agreed that ESOT's Executive Committee, Council, and Sections/Committees are diverse and inclusive (51%) and that ESOT promotes EDI in its live and digital educational activities (54%). Forty percent of respondents agreed that scientific and clinical trials in the field of transplantation are diverse and inclusive. Despite the wide distribution of the survey, most of the respondents self-identified as White and were either physician or surgeon. However, the results contribute a unique insight into the experiences and perspectives of the transplantation community regarding EDI. Whilst ESOT is committed to the principles of EDI, perceptions and the high number of proposals show the apparent need to prioritize efforts to embed EDI across ESOT and transplantation science. These data should constitute a starting point for change and provide guidance for future efforts to promote EDI within the transplantation community.

Abstract Objectives Neurokinin 2 receptor (NK2R) agonists may be useful for treating bladder and bowel dysfunction via direct contraction of detrusor and gastrointestinal smooth muscle. The NK2R agonist [Lys5, MeLeu9, Nle10]-NKA(4–10) (LMN-NKA) induces urination and defecation, but also produces the potential side effect of dermal flushing in rats. Although LMN-NKA is a NK2R agonist, it also has affinity for neurokinin 1 receptors (NK1R). Therefore, the goal of this study was to determine the neurokinin receptor (NKR) subtypes responsible for LMN-NKA-induced urination, defecation, and flushing by blocking either NK2Rs or NK1Rs before LMN-NKA administration. Methods To accomplish this goal, we developed a simple high-throughput ‘rapid detection voiding assay’ to detect rapid-onset drug-induced urination and defecation in rats. In LMN-NKA dose-response experiments, LMN-NKA (10–100 μg/kg, subcutaneous) was injected and urination, defecation, and flushing were monitored for 30 min. For NKR antagonist experiments, vehicle, the NK2R antagonist GR159897, or the NK1R antagonist CP-99,994 were injected before an acclimation period. Following acclimation, saline or 100 μg/kg LMN-NKA were injected, and behavior was observed for 30 min. Results LMN-NKA produced dose-related increases in urination, defecation, and flushing. Blocking NK2Rs reduced urination and blocked defecation, without affecting flushing. Blocking NK1Rs did not change LMN-NKA-induced urination or defecation but reduced LMN-NKA-induced flushing. Conclusions Using the rapid detection voiding assay we show that LMN-NKA-induced urination and defecation are mediated by NK2Rs, while flushing is mediated by NK1Rs. Therefore, drugs that are more selective for NK2 vs. NK1Rs should produce rapid-onset urination and defecation without producing the potential side effect of flushing.

Study design Animal proof of principle study. Objectives Bladder and bowel dysfunction are common after spinal cord injury (SCI) and in the elderly. Neurokinin 2 receptor agonists are known to produce on-demand urination and defecation in adult SCI rats. This study compared the ability of a neurokinin 2 receptor (NK2R) agonist to produce bladder and colorectal contractions in both young adult and aged SCI rats. Setting Dignify Therapeutics and Integrated Laboratory Systems, Durham, NC USA. Methods Bladder and colorectal pressure and voiding efficiency were measured in response to the NK2R agonist, [Lys5,Me,Leu9,Nle10]-NKA (4-10) (LMN-NKA), in anesthetized animals. The potency and efficacy of LMN-NKA was examined in young adult and aged SCI (T3 or T9 transected) rats, with young adult and aged spinal intact rats included as controls. Results LMN-NKA (3–300 μg/kg i.v.) produced dose-dependent increases in bladder and colorectal pressure in all anesthetized rats. No differences in the bladder or colorectal pressure responses or voiding efficiency were observed with age or after SCI. The level of SCI did not change the pharmacodynamic responses to the agonist. Conclusions An NK2R agonist produced similar responses in young adult and aged SCI rats, suggesting this class of agonists could be used as a potential therapy to induce on-demand urination and defecation in aged populations, with or without SCI.

Abstract Introduction MicroRNAs are promising biomarkers of renal disease, however the cellular origin of their expression is usually unclear limiting their interpretation when measured in renal biopsies and urine. We hypothesised that by first defining renal cell-enriched microRNAs, we could select biomarkers based on the expected histopathological profile. Method Small RNA-sequencing of cortical, proximal tubular (LTL), macrophage (F480), endothelial (CD31) and fibroblast (PDGFRb) populations from the reversible unilateral ureteric obstruction (rUUO) murine model was performed. Hierarchical clustering was used to identify clusters. Findings were translated into an ischaemia reperfusion injury (IRI) model and then into urine samples from renal transplant recipients (n=16) with delayed graft function (DGF) vs. those with primary function. Result Kidney injury resulted in significant macrophage infiltration and tubular injury which improved upon reversal. We characterised novel microRNA clusters enriched for each cell type. With injury there was a significant increase in macrophage (p<0.0001), fibroblast (p<0.01) and decrease in proximal tubule (p<0.0001) enriched microRNAs vs. non-enriched microRNAs. We validated macrophage enriched miR-18a, miR-16 and tubular enriched miR-194 in the IRI model, demonstrating that microRNA expression reflected the histological profile. In humans, urinary miR-16 (FC 16.9; p<0.05) and miR-18a (FC 10: p=0.06) were upregulated at day 2 in patients with DGF; outperforming the traditional injury marker KIM1. Conclusion This is the first study to characterise cell-enriched microRNAs during renal injury and repair. By defining the source of microRNA expression we were able to rationally select miR-16 and miR-18a as promising urinary biomarkers of renal injury. Take-home message We have found that microRNAs have differences in expression between cell types and renal injury states which is important when considering microRNA expression in samples composed of varying cellular composition. By defining the cellular origins of microRNA expression we were able to rationally select microRNA biomarkers of human renal injury.

Oxytocin has been shown to be important for social behavior and emotional attachments in early life and may also mediate effects of early experiences on social motivation in adulthood. In animal models, early maternal separation results in alterations in the oxytocin system, with effects on sexual, maternal, and stress reactivity behaviors in adulthood. Studies of children experiencing parental divorce find effects on mood disorders, substance abuse, and other behaviors in adulthood. Here, we examine the effect of divorce on adult urine oxytocin levels. To stimulate oxytocin release, participants, aged 18 to 62, were asked to complete a set of questionnaires on attachment style, parental history of divorce (age at parental divorce ranged from 0 to 20), and other measures. A sample of urine was then collected for the oxytocin assay. Urine oxytocin concentrations were substantially lower (p = .016) in subjects who experienced parental divorce (M = 3.70, Standard Error of the Mean = 0.73), compared to those who did not (M = 8.00, Standard Error of the Mean = 1.21), and correlated with responses on several attachment instruments. These results suggest that oxytocin levels are adversely affected by parental divorce in humans and may be related to attachment measures in adulthood. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Acute administration of [Lys5,Me,Leu9,Nle10]-NKA(4-10) (LMN-NKA) produces contractions of the detrusor and rectum with voiding in intact and acutely spinal cord injured (SCI) rats. In the current study, the ability of LMN-NKA (10 mu g/kg or 100 mu g/kg, subcutaneous [SC], twice a day [bid]) or vehicle to induce voiding and defecation in chronic SCI rats was examined across 30 days. After the last day of administration, voiding response rates and bladder pressure (BP) responses to LMN-NKA (intravenous [IV] and SC) were evaluated under anesthesia. In conscious rats, LMN-NKA (100 mu g/kg) produced dose-dependent micturition within 5 min, with response rates >90%, and voiding efficiency >80% in males and >60% in females, which remained stable across the 1-month test period. Similarly, LMN-NKA administration rapidly induced defecation, which also remained stable. Under anesthesia, LMN-NKA increased BP, voiding efficiency, and voiding response rates, which reached 100% at 3 and 10 mu g/kg IV in males and females, respectively. SC administration produced 100% response rates in males (30 mu g/kg) but only 71% in females (100 mu g/kg). Efficacy in rats chronically treated with LMN-NKA was similar to naive and vehicle-treated rats, except for reduced voiding efficiency in chronically dosed female rats (100 mu g/kg). No differences in bladder weights or collagen-to-smooth muscle ratios in histological sections were seen between the groups. Thus neither tolerance, nor sensitization, to LMN-NKA-induced micturition and defecation occurs with chronic administration in rats with chronic SCI. Efficacy was higher in male than in female rats.

The inability to initiate voluntary defecation affects many individuals with spinal cord injury (SCI).Current standard of care includes regular use of digital stimulation and manual extraction, laxatives, and/or enemas to prevent fecal impaction and/or incontinence.These bowel programs are time-consuming and stigmatizing, and fecal incontinence severely restricts individual productivity and quality-of-life.Restoration of voluntary defecation via a drug that produced a rapid, powerful, and brief contraction of the distal colonic and rectal smooth muscle, i.e. mimicked physiological contractions during defecation, would define a novel treatment.Neurokinin-2 receptor (NK2R) agonists were previously shown to produce highly potent and powerful contractions of sigmoid colon and rectal circular smooth, in vitro, in various species including humans.In vivo studies in spinal intact or chronic SCI rats indicated that the prototype NK2R agonist, LMN-NKA, can produce increased colorectal pressure and subsequent defecation within minutes of SC administration, with recovery to baseline pressure within 5-30 min, and no obvious adverse events at efficacious doses.LMN-NKA (30-100 ug/kg, SC) also increased colorectal pressure and induced defecation in dogs, but efficacious doses also produced a high rate of emesis in conscious dogs.Since dogs are extremely sensitive to emesis, compared to humans and other species, the current study examined efficacy and tolerability of LMN-NKA in minipigs with various routes of administration.In isoflurane-anesthetized minipigs instrumented to record rectal and arterial pressures, IV (0.3 ug/kg), SC (3-100 c ˇg/kg), IN (100 c ˇg/kg) and SL (5 mg/kg) administration (Table 1) produced similar increases in peak colorectal pressure (~35 mmHg) but with differing onset times.The effect was inhibited by a 15 min pretreatment with the NK2R antagonist GR 159897 (1 mg/kg; IV).Mean arterial pressure (MAP) was transiently decreased by 16% and 12% after SC and IV administration, respectively, which is NK1R-mediated in rats, dogs, and monkeys.MAP was not affected by IN or SL administration.In conscious minipigs (Table 2), 10-100 c ˇg/kg SC produced a dose-related increase in defecation responder rates with a 77% responder rate and no emesis at 30 ug/kg, while emesis was observed after 100 ug/kg.Both IN (100 c ˇg/kg) and SL (5 mg/kg) administration resulted in 80% defecation responder rates with no emesis observed.All routes of administration produced defecation in < 15 min.Repeated dosing at 30 ug/kg (SC), 3 times/day, for 5 consecutive days, in 6 pigs produced an 82% responder rate (41 events/50 trials) with no emesis (0 events/50 trials).These findings indicate that LMN-NKA causes less emesis in minipigs than dogs at doses that produce increases in colorectal pressure and rapid-onset, short-duration, ondemand defecation.

The effects of the neurokinin NK2 receptor agonist [Lys(5),MeLeu(9),Nle(10)]-NKA((4-10)) (LMN-NKA) on bladder and colorectal function were examined in minipigs. In anesthetized animals, subcutaneous (SC) administration of 30-100 mu g/kg increased peak bladder and colorectal pressures. Increases in bladder and colorectal pressure were inhibited by a 15 min pretreatment with the NK2 receptor antagonist GR 159897 (1 mg/kg intravenously (IV)). Bladder and colorectal pressures were also increased after IV (0.3 mu g/kg), intranasal (IN; 100 mu g/kg) and sublingual administration (SL; 5 mg/kg). There was a nonsignificant trend for hypotension (16 or 12% decrease in mean arterial pressure) after 100 mu g/kg SC and 0.3 mu g/kg IV, respectively, but not after 100 mu g/kg IN or 5 mg/kg SL. In conscious minipigs, 30-300 mu g/kg SC caused a dose-related increase in defecation that was accompanied by emesis in 38% of subjects receiving 300 mu g/kg. Urination was increased after 100 mu g/kg SC but not lower or higher doses. The peak plasma exposure (Cmax) after 100 mu g/kg SC was 123 ng/mL, and area under the curve (AUC) was 1790 min * ng/mL. Defecation response rates (similar to 82%) were maintained after SC administration of LMN-NKA (30 mu g/kg) given 3 times daily over 5 consecutive days. Defecation rates were higher after a single dose of 100 mu g/kg IN compared with vehicle, but this did not reach significance. After 7-10 mg/kg SL, 83% of animals urinated and defecated, and none had emesis. The data support the feasibility of developing a convenient and well-tolerated route of administration of LMN-NKA for human use. Minipigs may be a suitable species for toxicology studies with LMN-NKA due to the relatively low rate of emesis in this species.

Objective: To examine the safety and efficacy of using a clitoral vacuum suction device (CVSD) versus vibratory stimulation (V) to treat orgasmic dysfunction in women with multiple sclerosis (MS) or spinal cord injury (SCI). Design: Randomized clinical trial. Setting: Two academic medical centers. Participants: Women (N=31) including 20 with MS and 11 with SCI. Intervention: A 12-week trial of the use of a CVSD versus V. Main Outcome Measures: Female Sexual Function Inventory (FSFI) and Female Sexual Distress Scale (FSDS). Results: Twenty-three women (18 MS, 5 SCI) completed the study including 13 of 16 randomized to CVSD and 10 of 15 randomized to V. There was a statistically significant increase in total FSFI score (P=.011), desire (P=.009), arousal (P=.009), lubrication (P=.008), orgasm (P=.012), and satisfaction (P=.049), and a significant decrease in distress as measured by FSDS (P=.020) in subjects using the CVSD. In subjects who used V, there was a statistically significant increase in the orgasm subscale of the FSFI (P=.028). Subjects using the CVSD maintained improvements 4 weeks after treatment. Conclusions: CVSD is safe and overall efficacious to treat female neurogenic sexual dysfunction related to MS and SCI. V is also safe and efficacious for female neurogenic orgasmic dysfunction; however, results were limited to the active treatment period. Because of ease of access and cost, clinicians can consider use of V for women with MS or SCI with orgasmic dysfunction. CVSD is recommended for women with multiple sexual dysfunctions or for whom V is ineffective. (C) 2017 by the American Congress of Rehabilitation Medicine