INTRODUCTION Prostate cancer is the second most reported malig-nancy (after lung cancer) in men worldwide, with 1,276,106 new cases in 2018. Prostate cancer is respon- sible for 3.8% of all cancer deaths in men in men. 2018. (1) New cases of prostate cancer are estimated to emerge 2,293,818 cases by 2040, with projections of small var- iations in mortality in the form of a 1.05% increase.(2) About 40% of local prostate cancer patients decided to undergo radical prostatectomy.(3) Urinary incontinence is a typical and predictable post-radical prostatectomy episode and is triggered by activities such as sneezing, coughing, lifting, changing positions, and exercising. After prostatectomy, persis- tent and disturbing urinary incontinence is a common- ly reported side effect, with an incidence rate of 1% to 40% postoperatively.(4–6) However, this number might be much higher, depending on the definition used and the validity of the incontinence questionnaire used. Post-radical prostatectomy urinary incontinence sig- nificantly affects most men's quality of life undergoing surgical management of prostate cancer.(7) In assessing the quality of life of patients undergoing radical pros- tatectomy, incontinence was significantly associated with increased confusion, depression, and anger, and inversely related to physical and psychological health- iness.(8) As many as 28 and 18% of patients in the sur- gical group from the SPCG-4 study experienced mod- erate to severe discomfort due to urinary incontinence during the day or night.(9) Although improvements in surgical techniques have helped reduce the incidence of post-prostatectomy incontinence,(10) the overall rate continues to increase due to an increase in the total number of prostatectomies performed throughout the world. Several studies recommend delaying invasive urinary incontinence therapy at least one year postoperative- ly.(11,12) Therefore, behavioral therapy was chosen in several cases as an alternative.(13) This noninvasive be- havioral therapy consists of diet modification, bladder training, pelvic floor muscle training (PFMT ), biofeed- back, and functional electrical stimulation. In addition to being inexpensive and practical, they do not involve side effects.(14) However, some randomized controlled trials have investigated pelvic floor muscle training (PFMT) on postoperative urinary incontinence, provid- ing conflicting evidence. While some support PFMT exercise benefits,(15,16) a Cochrane review in 2015 does not recommend PFM training as a first-line rehabilita- tion performed post-prostatectomy because there are no significant improvements in UI symptoms over time.(17) The protocol related to PFM training initiated preop- eratively and continues postoperatively has not been established yet. The low level of evidence and the lack of systematic reviews that comprehensively review this technique's efficacy might be the contributing factor. On REVIEW The Effect of Preoperative Pelvic Floor Muscle Training on Incontinence Problems after Radical Prostatectomy: A Meta-Analysis Ervandy Rangganata1, Harrina Erlianti Rahardjo1* Purpose: To evaluate whether additional pelvic floow muscle training (PFMT), which began before radical prosta- tectomy and resumes immediately after catheter removal, will significantly improve urinary incontinence after RP. Materials and Methods: We reviewed articles obtained from MEDLINE, CENTRAL, EBSCOHost, CINAHL, and Elsevier from July – August 2020, which compared preoperative PFMT with postoperative PMFT or non-PF- MT, with continence incidence parameters. There were no restrictions on the definition of incontinence, treatment regimens, and radical prostatectomy surgical approach. The risk of bias was assessed using the Cochrane Risk of Bias Assessment Tool. A meta-analysis was also carried out to pool the effect estimates. Results: We included 12 eligible studies in this review, 11 of which we included in the meta-analysis. The PFMT initiated preoperatively significantly reduced the incidence of persistent urinary incontinence at 1, 3, and 6 months postoperatively with an OR of 0.58 (95% CI, 0.41–0.81), 0.57 (95% CI, 0.43–0.74), and 0.38 (95% CI, 0.17-0.83). There was no difference in improvement in patients' incontinence at 12 months postoperatively [OR = 1.31 (95% CI, 0.65-2.63)]. Conclusion: PFMT initiated before radical prostatectomy significantly reduced the incidence of urinary inconti- nence in the first, third, and sixth months postoperatively. At 12 months postoperatively, additional preoperative PFMT did not cause a significant difference in urinary incontinence incidence. Keywords: pelvic floor muscle training; urinary incontinence; radical prostatectomy 1Department of Urology, Cipto Mangunkusumo Hospital—Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia. *Correspondence: Department of Urology, Universitas Indonesia, Jakarta, Indonesia. Tel: +62 816 825226. E-mail: harrinaerlianti@gmail.com. Received September 2020 & Accepted April 2021 Urology Journal/Vol 18 No. 4/ July-August 2021/ pp. 380-388. [DOI: 10.22037/uj.v18i.6481] the other hand, several studies had shown a significant role for PFMT when it is initiated before surgery and/or early postoperatively (< 6 weeks postoperatively).(18,19) However, some studies also report controversial results. Therefore, the authors would like to evaluate whether a PFMT which begins before radical prostatectomy and resumes immediately after catheter removal, will sig- nificantly improve urinary incontinence. PATIENTS AND METHODS Description of condition and intervention This study was compiled based on the preferred re- porting items protocol for systematic reviews and me- ta-analysis (PRISMA) statements. This study attempted to explore the effectivity of PMFT initiated preoper- atively in improving urinary incontinence that occurs in patients after radical prostatectomy. Radical prosta- tectomy techniques were not limited to one particular approach. The use of laparoscopic and robot-assisted technology was also not a criterion for exclusion, as several studies have found there was no significant dif- ference between the type of prostatectomy technique to urinary incontinence.20,21 Improvement of urinary incontinence was not restricted to one definition or parameter. Besides, we also did not limit the types of interventions (PMFT) given to patients, other than the timeline of intervention that must be initiated before radical prostatectomy. A follow-up duration of at least three months was a requirement that must be fulfilled by each study. Database searching and literature screening We conducted literature searches using five search en- gines (Pubmed, Cochrane, EBSCOHost, ProQuest, and Scopus) based on four electronic databases (MEDLINE, CENTRAL, CINAHL, and EMBASE). The search was conducted from July 14 to August 1, 2020. We used PICOS in the literature screening process to assess the suitability of each study for this meta-analysis, as de- scribed on supp.Table 1. We used specific keywords, which were tailored to each search engine specification. We also looked at a reference list of several reviews to broaden the scope of study searches. Study selection Each author selected the study independently according to inclusion and exclusion criteria. Inclusion criteria in this study included: 1. RCT or quasi-RCT studies comparing PFMT (with or without biofeedback) before and after surgery with PMFT only after radical prostatectomy; 2. English/Indonesian written articles; 3. Full-text articles available; 4. Outcomes are the percentage of patients re- covering from incontinence; and 5. Were published in the last 20 years. We included all types of surgical techniques (open radi- cal prostatectomy, robot-assisted radical prostatectomy, and laparoscopic radical prostatectomy). The definition of incontinence of each study was also not a criterion for study selection. Exclusion criteria of this study in- cluded review articles, case reports, case series, edi- torial letters, studies on animals, and/or studies in the process of peer review (has not been published yet). The determination of study eligibility was determined by each author independently. After that, a full-text analysis was performed on the remaining article. Any disagreement was resolved by discussion. Data extraction and outcome of interest Each author conducted data extraction. We extracted the study's primary characteristics, including the first author, location, sample size, year of publication, and Figure 1. PRISMA flow chart describes the identification process of included articles. PFMT on Incontinence After RP-Rangganata et al. Review 381 patient's demography. We also extracted patient base- line data, including the degree of prostate malignan- cy and incontinence scores before the intervention (if available). We also extracted patterns or regimens of intervention (PMFT), but there were no restrictions on the pattern of PMFT. Any discrepancies have been re- solved by discussion. This meta-analysis explored the effectiveness of PMFT initiated before radical prostatectomy compared to PFMT initiated following radical prostatectomy or no PFMT at all, in improving the recovery rate of inconti- nence. The output of this study was the continence rate for each independent variable. We used 2x2 contingen- cy tabs to obtain the ORs for each study and pooled the overall OR using the Review Manager 5 application. Heterogeneity was measured by looking at the I2 value. The I2 value greater than 50% indicated a moderate to high heterogeneity. When the high heterogeneity was found between studies, an effect estimate analysis was performed with the DerSimonian and Laird random-ef- fects model. If heterogeneity between studies was low, fixed effect model analysis was performed using the Mantel-Haenszel methods. Assessment of methodologic quality This meta-analysis included only RCTs and qua- si-RCTs. RCT studies' quality assessment was carried out using the Cochrane risk-of-bias tool for randomized trials (RoB 2). Studies with, at least, moderate quality were included in the overall effect estimate calculation (meta-analysis). Red dot indicated a high risk of bias of each bias criterion, while yellow meant moderate and green meant low risk of bias. Selection bias criterion was not applicable for quasi-RCT study as no random sequence was generated in patient selection. Blinding of participant was also impossible in these settings. RESULTS Literature search A literature search on five electronic databases found 883 articles (first hit), and we found 285 similar arti- cles. For 598 remaining articles, we screened and found that only 17 articles were eligible. We independently conducted a full-text analysis of the remaining 17 ar- ticles and found five articles that did not fit the PICOS that we specified in this meta-analysis. We included 12 articles in this review and 11 articles in quantitative synthesis (meta-analysis)—flowchart of this literature search described in Figure 1. Study characteristics We found 12 eligible studies based on the suitability of patient characteristics, types of interventions in the experimental and control groups, and outcomes meas- ured in each study (Table 1). Eleven studies were ran- domized controlled trials and only 1 study with a qua- si-RCT design, which did not implement randomization and patient’s data was fully based on the patient's medi- cal record. Five studies lasted for six months, four stud- ies lasted for 12 months, and only three studies lasted for three months. Overall, the total number of patients involved in this study was 1348 patients. 3 of the 12 studies applied the same treatment regimen between the control and experimental groups,(22–24) two studies did not apply any treatment to the control group,(25,26) and the rest applied different treatment regimens. (18,19,27–31) Risk of bias of included studies Each author assessed each study's quality using the Cochrane Risk of Bias Assessment (RoB) tool for RCT independently. Overall, studies had a high risk of per- formance bias caused by the impossibility of partici- pants and personnel blinding (Figure 2). Therefore, we did not exclude studies just because of the high risk of Figure 2. Risk of bias assessment of RCTs using Cochrane Risk of Bias Assessment PFMT on Incontinence After RP-Rangganata et al. Vol 18 No 4 July-August 2021 382 performance bias. In general, the studies included in this meta-analysis were of good quality. The selection bias from the study of Patel et al. 2013 could not be as- sessed because this study was a quasi-RCT study. Intervention regimen We found a large variety of interventions given to pa- tients in each study, which was generally based on each health center's protocol, where the study was conducted. PFMT accompanied and guided by a physiotherapist, followed by a biofeedback session, was the most widely Review 383 Figure 3. Forest plot shows odds ratio of getting incontinence after radical prostatectomy at several time points: (A) 1 month; (B) 3 months; (C) 6 months; and (D) 12 months. Figure 4. Funnel plot shows the distribution of studies based on the effect estimate and standard error: (A) 1 month; (B) 3 months; (C) 6 months; and (D) 12 months. PFMT on Incontinence After RP-Rangganata et al. used treatment regimen.(18,22,24,25) Only the Parekh et al. (2003) study did not include biofeedback in the treat- ment regimen.(29) Most studies conducted training and biofeedback sessions at least once a week for 30–60 minutes per session, four weeks before radical prosta- tectomy. Only the Tienforti et al.(30) and Sayilan et al.(26) studies conducted therapy less than two weeks before radical prostatectomy, and two studies did not report the specific time of the PFMT therapy initiation.(19,29) Incontinence rate at several time points We compiled incontinence rates between studies in the form of OR patients experience post-radical prosta- tectomy incontinence at several time points (Table 2 and Figure 3). In the first 1-month after radical pros- tatectomy, we found that the experimental group had a significantly lower risk for incontinence after radical prostatectomy. The significance of this risk difference is was marked by the low OR and the width of the con- fidence interval that does did not exceed one unit [OR = 0.58 (95% CI, 0.41–0.81)]. The heterogeneity between studies was also not significant (I2 = 44%, p = 0.09), so we performed calculations using the Mantel-Haenszel fixed-effect model. At three months postoperatively, the incontinence rate’s difference between the experimental and control groups also remained significant. The experimental group's odds ratio for incontinence compared to the control group at three months postoperatively was 0.57 (95% CI, 0.43–0.74), with no significant heterogeneity be- tween studies (I2 = 48%, p = 0.05). Only two studies reported that the control group reported a lower risk of persistent incontinence than the experimental group. (23,27) Six months postoperatively, the incidence of urinary incontinence was still significantly lower in the ex- perimental group. At this time point, the experimental group's odds ratio to the control group was 0.38 (95% CI, 0.17–0.83), with significant heterogeneity between studies (I2 = 57%, p = 0.03). Only seven studies com- pared incontinence in the experimental and control groups in the 6th-month post radical prostatectomy, and only two studies reported a significantly higher in- cidence of persistent urinary incontinence six months postoperatively in the control group. The minimum number of studies involved in the meta-analysis for this six-month time point and the considerable variation in odds ratios between studies might become the cause of high heterogeneity between studies. There were only three studies comparing incontinence Author Intervention* Timing Control† N Age (yr) Duration Bales et al. 2000.27 Nurse guided graded PFMT 4 times/day Oral and verbal advice 47/50 59.3/60.9 6 months with biofeedback 2–4 weeks to surgery to perform PFMT without biofeedback Burgio et al. 2006.19 Instruction of PFMT with single NR Verbal advice to perform 57/55 60.7 ± 6.6/ 6 months session of biofeedback PFMT 60.7 ± 6.6 Centemero et al. Physiotherapist guided PFMT Twice per week Verbal instruction of 59/59 60.5 (48–68) 3 months 2010.18 with visual feedback (30 minutes), 2–4 PFMT with visual 57.5 (46–67) weeks preoperatively feedback / Collado et al. Written instruction of PFMT 3 weeks preoperatively Verbal instruction of 87/92 NR 12 months 2013.28 with weekly assisted-biofeedback Kegel exercise session and TVA Dijkstra-Eshuis et al. Physiotherapist guided PFMT with 30 minutes weekly, Physiotherapist guided 65/56 63.7 ± 5.3 12 months 2015.22 biofeedback and ES 4 weeks preoperatively PFMT with biofeedback and ES Geraerts et al. Guided PFMT with digital/EMG 30 minutes weekly, Guided PFMT with 85/85 61.88 (44–73)/12 months 2013.23 biofeedback 3 weeks preoperatively digital/EMG biofeedback 62.04 (41–76) Lira et al. 2019.25 Physiotherapist guided PFMT with 3 times/day 3 weeks Postoperative care 16/15 63.53 ± 7.62/ 3 months EMG biofeedback preoperatively without PFMT 67.3 ± 5.63 Parekh et al. 2003.29 Formal instruction with 2 sessions of 2 sessions preoperatively Non-formal PFMT 19/19 55.5/61.6 12 months guided PFMT instruction Patel et al. 2013.24 Physiotherapist guided PFMT with 1–4 sessions (1 hour), Physiotherapist guided 152/132 60 (41–76)/ 3 months visual feedback 4 weeks preoperatively PFMT with visual 62 (44–76) feedback Sayilan et al. 2018.26 Guided PFMT with visual feedback 1–4 sessions (1 hour), No exercise instruction 30/30 63.00 ± 8.61/ 6 months 1 week preoperatively 59.93 ± 6.98 Tienforti et al. Supervised PFMT with biofeedback 1 day preoperatively Oral and written 16/16 67 (60–74)/ 6 months 2012.30 instructions of PFMT 64 (52–74) Yoshida et al. Physician guided PMFT with single 1 month preoperatively Verbal instruction of 36/80 66.5 ± 6.2/ 6 months 2018.31 session US-biofeedback PFMT 66.5 ± 5.8 Table 1. Characteristic of the study included in this systematic review. * Interventions were given both before and after surgery; † Interventions were given postoperatively only; NR: not reported; TVA: trans- versus abdominis activation; ES: electrical stimulation; PFMT: pelvic floor muscle training; EMG: electromyographic; US: ultrasound PFMT on Incontinence After RP-Rangganata et al. Vol 18 No 4 July-August 2021 384 events at 12 months post radical prostatectomy.(22,23,29) None of these studies reported significant differences between incontinence events in the experimental and control groups. Therefore, we also found something similar in the meta-analysis at that time point. We found no significant difference between the control and experimental groups on experiencing incontinence 12-month after radical prostatectomy, with the experi- mental group's tendency to be more susceptible [OR = 1.31 (95% CI, 0.65-2.63), I2 = 30%, p = 0.44]. We used a funnel plot to predict the probability of publication bias in this meta-analysis (Figure 4). The risk of publication bias was relatively high in several outcomes: six months and 12 months after surgery. The low number of studies assessing these outcomes, especially 12 months postoperatively, variations in pa- tient characteristics and outcome measurement, and the inclusion of studies with low quality were thought to cause the high risk of publication bias. Subgroup analysis was not carried out as we did not find any relevant characteristic that might influence the outcome. The types of prostatectomy (ORP, LARP, RARP) did not significantly affect the incontinence rate after prostatectomy.(20,21) DISCUSSION Persistent and disturbing urinary incontinence after prostatectomy is a commonly reported side effect post- operatively, with an incidence rate of 1% to 40%.(4–6) Several studies recommend delaying invasive urinary incontinence therapy at least one year postoperatively. (11,12) Therefore, behavioral therapy was chosen in some cases as an alternative.(13) This noninvasive behavioral therapy consists of diet modification, bladder training, pelvic floor muscle training (PFMT), biofeedback, and functional electrical stimulation. Apart from being cheap and practical, these therapies have never been re- ported to cause any side effects.(14) Urinary continence depends on the smooth and striated muscle fibers' complex interactions that work sinergi- cally to form a continuity mechanism. Some authors are still debating about whether incontinence after prosta- tectomy is due to effects on the detrusor muscle (blad- der) or the sphincter. Detrusor overactivity and intrinsic sphincter insufficiency due to sphincteric injury are the most important causes of persistent incontinence after radical prostatectomy. Some reports mention that detru- sor overactivity is a significant cause of postprostatec- tomy incontinence,(32,33) others strongly argue that even if other factors play a role, intrinsic sphincter deficiency is the main cause of UI after radical prostatectomy.(34–36) Detrusor overactivity is an incontinence pathophysiol- ogy that is corrected by PFMT. This method includes exercising specific pelvic floor voluntary muscle con- tractions using biofeedback, as well as coordinating and determining the time of contraction for increased in- traabdominal pressure. Specific and repetitive contrac- tions of the pelvic floor muscles can increase strength and efficiency when there is an increase in intraabdom- inal pressure; thus, this would suppress detrusor over- activity. Berghmans and colleagues reported that pelvic floor muscle contractions effectively held the urethra by Review 149Review 385 Author n Continence definition Types of prostatectomy Incontinence (OR) Quality 1 mo 3 mo 6 mo 12 mo Bales et al. 2000 47/50 The use of ≤ 1 pad per day NR 1.33 1.21 1.64 NR Fair Burgio et al. 2006 57/55 No leakage (3 consecutive NR 0.19 0,37 0.48 NR Fair weekly 1-day diaries/7- day diary) Centemero et al. 2010 59/59 No urinary leakage in ORP 0.32 0.41 NR NR Fair bladder diary and a negative stress test Collado et al. 2013 87/92 Not reported NR NR NR NR NR Fair Dijkstra-Eshuis et al. 65/56 No leakage at all on a LARP NR NR NR 2.11 Fair 2015 24-hr pad test, PeLFIs, and KHQ Geraerts et al. 2013 85/85 3 days of 0 g of urine RARP and ORP 1.00 1.36 1.00 0.49 Fair loss on the 24-h pad test Lira et al. 2019 16/15 Patient’s perception of ORP NR 0.80 NR NR Good loss of at least a few drops of urine Parekh et al. 2003 19/19 The use of ≤ 1 pad per day NR 0.46 0.27 NR 0.79 Fair Patel et al. 2013 152/132 Patient-reported one pad ORP NR 0.61 NR NR Poor usage/day Sayilan et al. 2018 30/30 ICIQ-UI score of zero NR 0.29 0.11 0.03 NR Good Tienforti et al. 2012 16/16 ICIQ-UI score of zero ORP 0.05 0.07 0.04 NR Good Yoshida et al. 2018 36/80 The number of days NR 0.49 0.48 0.40 NR Good requiring a small pad (20 g)/day Table 2. Systematic review table of the experiment’s outcomes. NR: not reported, ORP: open radical prostatectomy, RARP: robot-assisted laparoscopic radical prostatectomy, LARP: laparoscopic rad- ical prostatectomy PFMT on Incontinence After RP-Rangganata et al. providing structural support to the pelvic organs; and PFMT triggers hypertrophy of the urethral muscular muscles thereby increasing mechanical pressure on the urethra, which in turn can prevent detrusor overactivity and prevent urinary incontinence.(37) Several clinical studies have proven that the strength of PFM correlates with incontinence and that PFMT in- creases the strength of PFM can effectively speed up the recovery of incontinence in patients post radical prostatectomy.(17,38) In this study, we found that PFMT carried out before radical prostatectomy significantly reduced the risk of persistent urinary incontinence one month after radical prostatectomy [OR = 0.58 (95% CI, 0.41–0.81)], compared to patients who underwent PFMT only after surgery or did not undergo PFMT at all. This reduction in incontinence risk is consistent for up to 6 months postoperatively, in which most studies agree that the experimental group has a much higher rate of continence than the control group. At 12 months postoperatively, the control group could achieve the same continence rate as the experimental group, indi- cating that almost all patients in both groups had re- gained continence at 12 months postoperatively [OR = 1.31 (95% CI, 0.65–2.63)]. This meta-analysis combined several radical prostatec- tomy approaches, such as open radical prostatectomy (ORP), laparoscopic radical prostatectomy (LARP), and robot-assisted (RARP). A combination of these approaches was undertaken to increase the heterogene- ity of studies extrapolated in various clinical settings. Moreover, several prospective comparative studies found no statistically significant difference in urinary incontinence between post-ORP, LRP, or RARP pa- tients.(20,21) Our findings are in line with Chang et al. (2016), who included six studies in their meta-analysis to determine the effect of PFMT in improving incontinence. The study found that the experimental group's odds were lower in 1, 3, and 6 months postoperatively and were significantly different in the three months postopera- tively.(39) Some previous meta-analyses reported the opposite of what we found. Wang et al. (2014) reported no significant difference in the incidence of the relative risk of persistent incontinence at 1, 3, 6, and 12 months post radical prostatectomy.(40) However, this study in- cluded no more than four studies for meta-analysis, and there were only two studies that have good quality. A meta-analysis by Wu et al. (2019) found that guided-PF- MT effectively reduced the risk of persistent inconti- nence at all time points, and preoperative guide-PFMT did not provide any benefit to patients. However, this study only included two studies and two-time points (3 and 6 months), and had a high heterogeneity (79%).(41) The impact of incontinence on a patient's quality of life is clearly visible. Geraerts et al. (2013) reported a smaller reduction in quality of life in the preoperative exercise group, and all patients in the experimental group expressed satisfaction in receiving PFMT before surgery.(23) Research by Centemero et al. (2010) showed that 75% of patients in the intervention group reported a high level of satisfaction by starting PFMT before sur- gery.(18) Considering that urinary incontinence is a com- plication that significantly reduces the quality of life, any intervention that can shorten its duration is worth a try. Moreover, patients who receive additional PFMT before surgery show a high level of satisfaction, so PMFT can be a noninvasive therapy option that should be recommended to patients before radical prostatecto- my. There are several limitations in this meta-analysis that may interfere with the interpretation of final results. We found considerable heterogeneity between studies. This heterogeneity arises due to the large variety of PFMT regimens in each study. For example, PFMT accompa- nied and guided by a physiotherapist, accompanied by a biofeedback session, is the most widely used treatment regimen, but only 4 of 12 studies used this treatment regimen. One included study did not even include bi- ofeedback in the treatment regimen. The definition of the "intervention group" in each study also varies. Three studies considered the intervention as an addi- tional PFMT so that the treatment regimen between the control and experimental groups was the same, with the only difference being the time the therapy was started. Moreover, the definition of incontinence and conti- nence of each study was also diverse, and it was impos- sible to establish a single definition of incontinence as an inclusion criterion. Some other things that cause high heterogeneity were surgery techniques, the frequency of PFMT intervention, and the time of initiation of pre- operative and postoperative PFMT. Our meta-analysis included quite a large number of studies, and most had a low risk of bias, considering that participant blinding was not possible in this study with PICO. CONCLUSIONS PFMT initiated before radical prostatectomy signifi- cantly reduced the incidence of urinary incontinence in the first, third, and sixth months postoperatively. At 12 months postoperatively, additional preoperative PFMT did not cause a significant difference in the incidence of urinary incontinence. CONFLICT OF INTEREST The authors report no conflict of interest. ACKNOWLEDGMENTS The first author of the paper (E. 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