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MP21-19 THE IMPACT OF AUTHOPHAGY IN NEUROPATHIC BLADDER REMODELING
THE JOURNAL OF UROLOGYâ
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MP21-19 THE IMPACT OF AUTHOPHAGY IN NEUROPATHIC BLADDER REMODELING Souzan Salemi, Ashkan Mortezavi, Maya Horst, Rita Gobet, € rich, Switzerland Tullio Sulser, Daniel Eberli*, Zu INTRODUCTION AND OBJECTIVES: Neurogenic bladder dysfunction is the result of disease or injury to the neural pathways and commonly occurs in patients with meningomyelocele or after spinal cord injury. During the pathogenesis the smooth muscle cells (SMC) shift from contractile SMC phenotype towards a synthetic type. In muscular disorders increased autophagy is known to protect cells from deterioration by compensating for defects in lysosome function. However, the accumulation of autophagosomes can also impair cell function over time. Autophagy may play an important role in remodeling of bladder SMC in children with neuropathic bladder. In this study we investigated the role of autophagy in neuropathic bladders in the pediatric population. METHODS: Full thickness bladder biopsies were taken from children with neuropathic disorder. Samples obtained from healthy donors without urological problems were used as control. A piece of bladder tissue was snap frozen for genetic analysis and another piece was fixed for immunostaining. Samples were stained with SMC lineage -SMA and autophagy proassociated markers calponin, smoothelin, a teins LC3, Atg5 and Beclinl. In addition the expression of autophagy genes and proteins were investigated by real time PCR and Western blot analysis. RESULTS: We found that the ATG5 gene, a key regulator of autophagy, is upregulated in neuropathic muscle tissue compared with normal bladder. At protein level increased ATG5 protein was repetitively shown in WB and immunostaining. Neuropathic bladder muscle exhibited a punctated immunostaining pattern for LC3 in subset of SM confirming accumulation of autophagosomes. Pronounced elevation of ATG5 in SM in neuropathic bladder tissue co-localized with a downregulation of the key contractile proteins smoothelin and calponin. CONCLUSIONS: Our study reveals that autophagy is important factor in the remodeling of SMC and functionality of bladder SM tissue in neuropathic bladder. Since autophagy can be influenced by oral medication this research might lead to novel strategies preventing the remodeling and deterioration of neuropathic bladder muscle tissues. Source of Funding: none
MP21-20 INTRAVESICAL INHIBITION OF P2X3 RECEPTORS IMPROVES BLADDER DYSFUNCTION DEPENDING ON UROTHELIAL EXPRESSION LEVELS FOR THE RECEPTOR IN SCI RATS Jorge Tovar-Perez, Broderick Sutton, Timothy Boone, Alvaro Munoz*, Houston, TX INTRODUCTION AND OBJECTIVES: P2X3-purinergic receptors (P2X3R) are expressed in afferent nerve terminals of the urinary bladder where ATP released from the urothelium during filling increase nerve activity to begin the micturition reflex. P2X3R are also expressed in the urothelium, however, its physiological role is less clear. Our objectives were 1) to determine the cystometric effects of AF-353, a specific P2X3R antagonist, in intact animals and spinal cord injured (SCI) rats with neurogenic bladder; and 2) to compare the differences on urothelial P2X3R expression between groups. METHODS: Neurogenic bladder conditions were induced in female Sprague-Dawley rats via a partial bilateral-transection of the dorsal spinal cord at the Th8/Th9 region. Intact rats were used as controls. At 2 (SCI2W) or 4 weeks (SCI4W) post-SCI, rats were anesthetized with urethane and implanted with a suprapubic catheter for cystometry. Saline infusion was started at a rate of 0.1 ml/min. After baseline recordings (60e90 minutes), infusion containing AF-353 (10 mM) was initiated for a similar period of time. Thereafter, rats were perfused with iced-cold paraformaldehyde (4%). The bladder was
Vol. 193, No. 4S, Supplement, Saturday, May 16, 2015
isolated for immunostaining with a P2X3R antibody (abcam; 1:1,000) and DAPI. Confocal images were obtained at 40X. The bladder peak pressure (BPP), intercontractile intervals (ICI), and the frequency of voiding (VC) and non-voiding (NVC) contractions were determined. Changes were normalized with the respect to the saline-infusion values. RESULTS: Inhibition of P2X3R had minor effects on increasing the ICI in SCI2W rats, but significantly increased the ICI value in control and SCI4W rats. Application of AF-353 also decreased the frequency of NVC in both the SCI2W and SCI4W animals. The qualitative evaluation using confocal microscopy suggests that in comparison with the control bladders, the expression of P2X3R is reduced in SCI2W but increased in SCI4W rats. CONCLUSIONS: Our results suggest that the reduced effect of AF-353 to increased ICI in SCI2W rats may be related to a reduced expression of urothelial P2X3R. It is also reasonable to suggest that urothelial P2X3R may play a role in regulating ICI and the frequency of NVC in SCI rats. Pharmacological inhibition of P2X3R to treat neurogenic bladder conditions may be a beneficial therapy but requires taking into consideration the duration of the dysfunction. Source of Funding: Houston Methodist Foundation, the Cullen Foundation and the Brown Foundation.
Technology & Instruments: Surgical Education & Skills Assessment I Moderated Poster 22 Saturday, May 16, 2015
10:30 AM-12:30 PM
MP22-01 JOURNAL CLUBS VIA MICROBLOGGING ARE GLOBALIZING CONTINUING MEDICAL EDUCATION: A SYSTEMATIC REVIEW Matthew Roberts*, Brisbane, Australia; Marlon Perera, Mackay, Australia; Nathan Lawrentschuk, Diana Romanic, Nathan Papa, Damien Bolton, Melbourne, Australia INTRODUCTION AND OBJECTIVES: Journal clubs (JCs) are an essential tool in promoting clinical evidence-based medicine to all medical and allied health professionals. Twitter represents a public, microblogging forum that can facilitate traditional JC requirements, while also reaching a global audience and participation for discussion with study authors and colleagues. The objective of the current study was to evaluate the current state of JC facilitated by social media, specifically Twitterâ, as an example of CME and through a systematic review describe progress to date. METHODS: A systematic review of literature databases (Medline, Embase, CINAHL, Web of Science, ERIC via ProQuest) was performed according to PRISMA guidelines. All manuscripts related to Twitter-based JCs were included. A systematic search of Twitter, the followers of identified JCs, and Symplur.com, was also performed. Demographic and monthly tweet data were extracted from Twitter and Symplur. RESULTS: From a total of 469 citations, eleven manuscripts were included and referred to five Twitter-based JCs (#ALiEMJC, #BlueJC, #ebnjc, #iurojc, #meded). Hashtag and Twitter-based JC search yielded 34 potential hashtags/accounts were collated and 24 included in the final analysis, which had been active for a median of 11.75 (range 1e40, SD 10.9) months with seven now inactive. The median number of followers was 374 (range 72e3,446, SD 1,019) and 171 (range 42 to 1,954, SD 516), respectively. There is an overall increasing establishment of active Twitter-based JCs, resulting in an exponential increase in total cumulative tweets (R2 ¼ 0.98), and tweets per month (R2 ¼ 0.72). Cumulative tweets for specific JCs increased linearly, with @NephJC, @igsjc, @EBNursingBMJ, @iurojc and @ADC_JC showing greatest rate of change since establishment and total impressions per month. Tweets per month for individual JC’s
e240
MP21-19 THE IMPACT OF AUTHOPHAGY IN NEUROPATHIC BLADDER REMODELING Souzan Salemi, Ashkan Mortezavi, Maya Horst, Rita Gobet, € rich, Switzerland Tullio Sulser, Daniel Eberli*, Zu INTRODUCTION AND OBJECTIVES: Neurogenic bladder dysfunction is the result of disease or injury to the neural pathways and commonly occurs in patients with meningomyelocele or after spinal cord injury. During the pathogenesis the smooth muscle cells (SMC) shift from contractile SMC phenotype towards a synthetic type. In muscular disorders increased autophagy is known to protect cells from deterioration by compensating for defects in lysosome function. However, the accumulation of autophagosomes can also impair cell function over time. Autophagy may play an important role in remodeling of bladder SMC in children with neuropathic bladder. In this study we investigated the role of autophagy in neuropathic bladders in the pediatric population. METHODS: Full thickness bladder biopsies were taken from children with neuropathic disorder. Samples obtained from healthy donors without urological problems were used as control. A piece of bladder tissue was snap frozen for genetic analysis and another piece was fixed for immunostaining. Samples were stained with SMC lineage -SMA and autophagy proassociated markers calponin, smoothelin, a teins LC3, Atg5 and Beclinl. In addition the expression of autophagy genes and proteins were investigated by real time PCR and Western blot analysis. RESULTS: We found that the ATG5 gene, a key regulator of autophagy, is upregulated in neuropathic muscle tissue compared with normal bladder. At protein level increased ATG5 protein was repetitively shown in WB and immunostaining. Neuropathic bladder muscle exhibited a punctated immunostaining pattern for LC3 in subset of SM confirming accumulation of autophagosomes. Pronounced elevation of ATG5 in SM in neuropathic bladder tissue co-localized with a downregulation of the key contractile proteins smoothelin and calponin. CONCLUSIONS: Our study reveals that autophagy is important factor in the remodeling of SMC and functionality of bladder SM tissue in neuropathic bladder. Since autophagy can be influenced by oral medication this research might lead to novel strategies preventing the remodeling and deterioration of neuropathic bladder muscle tissues. Source of Funding: none
MP21-20 INTRAVESICAL INHIBITION OF P2X3 RECEPTORS IMPROVES BLADDER DYSFUNCTION DEPENDING ON UROTHELIAL EXPRESSION LEVELS FOR THE RECEPTOR IN SCI RATS Jorge Tovar-Perez, Broderick Sutton, Timothy Boone, Alvaro Munoz*, Houston, TX INTRODUCTION AND OBJECTIVES: P2X3-purinergic receptors (P2X3R) are expressed in afferent nerve terminals of the urinary bladder where ATP released from the urothelium during filling increase nerve activity to begin the micturition reflex. P2X3R are also expressed in the urothelium, however, its physiological role is less clear. Our objectives were 1) to determine the cystometric effects of AF-353, a specific P2X3R antagonist, in intact animals and spinal cord injured (SCI) rats with neurogenic bladder; and 2) to compare the differences on urothelial P2X3R expression between groups. METHODS: Neurogenic bladder conditions were induced in female Sprague-Dawley rats via a partial bilateral-transection of the dorsal spinal cord at the Th8/Th9 region. Intact rats were used as controls. At 2 (SCI2W) or 4 weeks (SCI4W) post-SCI, rats were anesthetized with urethane and implanted with a suprapubic catheter for cystometry. Saline infusion was started at a rate of 0.1 ml/min. After baseline recordings (60e90 minutes), infusion containing AF-353 (10 mM) was initiated for a similar period of time. Thereafter, rats were perfused with iced-cold paraformaldehyde (4%). The bladder was
Vol. 193, No. 4S, Supplement, Saturday, May 16, 2015
isolated for immunostaining with a P2X3R antibody (abcam; 1:1,000) and DAPI. Confocal images were obtained at 40X. The bladder peak pressure (BPP), intercontractile intervals (ICI), and the frequency of voiding (VC) and non-voiding (NVC) contractions were determined. Changes were normalized with the respect to the saline-infusion values. RESULTS: Inhibition of P2X3R had minor effects on increasing the ICI in SCI2W rats, but significantly increased the ICI value in control and SCI4W rats. Application of AF-353 also decreased the frequency of NVC in both the SCI2W and SCI4W animals. The qualitative evaluation using confocal microscopy suggests that in comparison with the control bladders, the expression of P2X3R is reduced in SCI2W but increased in SCI4W rats. CONCLUSIONS: Our results suggest that the reduced effect of AF-353 to increased ICI in SCI2W rats may be related to a reduced expression of urothelial P2X3R. It is also reasonable to suggest that urothelial P2X3R may play a role in regulating ICI and the frequency of NVC in SCI rats. Pharmacological inhibition of P2X3R to treat neurogenic bladder conditions may be a beneficial therapy but requires taking into consideration the duration of the dysfunction. Source of Funding: Houston Methodist Foundation, the Cullen Foundation and the Brown Foundation.
Technology & Instruments: Surgical Education & Skills Assessment I Moderated Poster 22 Saturday, May 16, 2015
10:30 AM-12:30 PM
MP22-01 JOURNAL CLUBS VIA MICROBLOGGING ARE GLOBALIZING CONTINUING MEDICAL EDUCATION: A SYSTEMATIC REVIEW Matthew Roberts*, Brisbane, Australia; Marlon Perera, Mackay, Australia; Nathan Lawrentschuk, Diana Romanic, Nathan Papa, Damien Bolton, Melbourne, Australia INTRODUCTION AND OBJECTIVES: Journal clubs (JCs) are an essential tool in promoting clinical evidence-based medicine to all medical and allied health professionals. Twitter represents a public, microblogging forum that can facilitate traditional JC requirements, while also reaching a global audience and participation for discussion with study authors and colleagues. The objective of the current study was to evaluate the current state of JC facilitated by social media, specifically Twitterâ, as an example of CME and through a systematic review describe progress to date. METHODS: A systematic review of literature databases (Medline, Embase, CINAHL, Web of Science, ERIC via ProQuest) was performed according to PRISMA guidelines. All manuscripts related to Twitter-based JCs were included. A systematic search of Twitter, the followers of identified JCs, and Symplur.com, was also performed. Demographic and monthly tweet data were extracted from Twitter and Symplur. RESULTS: From a total of 469 citations, eleven manuscripts were included and referred to five Twitter-based JCs (#ALiEMJC, #BlueJC, #ebnjc, #iurojc, #meded). Hashtag and Twitter-based JC search yielded 34 potential hashtags/accounts were collated and 24 included in the final analysis, which had been active for a median of 11.75 (range 1e40, SD 10.9) months with seven now inactive. The median number of followers was 374 (range 72e3,446, SD 1,019) and 171 (range 42 to 1,954, SD 516), respectively. There is an overall increasing establishment of active Twitter-based JCs, resulting in an exponential increase in total cumulative tweets (R2 ¼ 0.98), and tweets per month (R2 ¼ 0.72). Cumulative tweets for specific JCs increased linearly, with @NephJC, @igsjc, @EBNursingBMJ, @iurojc and @ADC_JC showing greatest rate of change since establishment and total impressions per month. Tweets per month for individual JC’s