Chemical modulation of SQSTM1/p62-mediated xenophagy that targets a broad range of pathogenic bacteria.

The N-degron pathway is a proteolytic system in which the N-terminal degrons (N-degrons) of proteins, such as arginine (Nt-Arg), induce the degradation of proteins and subcellular organelles via the ubiquitin-proteasome system (UPS) or macroautophagy/autophagy-lysosome system (hereafter autophagy). Here, we developed the chemical mimics of the N-degron Nt-Arg as a pharmaceutical means to induce targeted degradation of intracellular bacteria via autophagy, such as Salmonella enterica serovar Typhimurium (S. Typhimurium), Escherichia coli, and Streptococcus pyogenes as well as Mycobacterium tuberculosis (Mtb). Upon binding the ZZ domain of the autophagic cargo receptor SQSTM1/p62 (sequestosome 1), these chemicals induced the biogenesis and recruitment of autophagic membranes to intracellular bacteria via SQSTM1, leading to lysosomal degradation. The antimicrobial efficacy was independent of rapamycin-modulated core autophagic pathways and synergistic with the reduced production of inflammatory cytokines. In mice, these drugs exhibited antimicrobial efficacy for S. Typhimurium, Bacillus Calmette-Guérin (BCG), and Mtb as well as multidrug-resistant Mtb and inhibited the production of inflammatory cytokines. This dual mode of action in xenophagy and inflammation significantly protected mice from inflammatory lesions in the lungs and other tissues caused by all the tested bacterial strains. Our results suggest that the N-degron pathway provides a therapeutic target in host-directed therapeutics for a broad range of drug-resistant intracellular pathogens.Abbreviations: ATG: autophagy-related gene; BCG: Bacillus Calmette-Guérin; BMDMs: bone marrow-derived macrophages; CALCOCO2/NDP52: calcium binding and coiled-coil domain 2; CFUs: colony-forming units; CXCL: C-X-C motif chemokine ligand; EGFP: enhanced green fluorescent protein; IL1B/IL-1ß: interleukin 1 beta; IL6: interleukin 6; LIR: MAP1LC3/LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; Mtb: Mycobacterium tuberculosis; MTOR: mechanistic target of rapamycin kinase; NBR1: NBR1 autophagy cargo receptor; OPTN: optineurin; PB1: Phox and Bem1; SQSTM1/p62: sequestosome 1; S. Typhimurium: Salmonella enterica serovar Typhimurium; TAX1BP1: Tax1 binding protein 1; TNF: tumor necrosis factor; UBA: ubiquitin-associated.

Rg6, a rare ginsenoside, inhibits systemic inflammation through the induction of interleukin-10 and microRNA-146a.

The immunobiological functions of Rg6, a rare ginsenoside from ginseng, have been largely unreported. In this paper, we demonstrate that Rg6 has a significant immunosuppressive function on Toll-like receptor (TLR) 4-induced systemic inflammatory responses. Rg6 was found to negatively regulate pro-inflammatory responses and severity in vivo, and thus induced recovery in mice with lipopolysaccharide (LPS)-induced septic shock and cecal ligation and puncture (CLP)-induced sepsis. Rg6 treatment also facilitated recovery in mice with LPS-induced lung damage via reduced neutrophil infiltration and tumor necrosis factor-α expression in lung tissues. Rg6 injection also downregulated pro-inflammatory cytokines and increased the levels of interleukin (IL)-10 in the serum of septic mice. Mechanistically, Rg6 did not induce TLR negative regulators, such as A20 and IRAK-M, in bone marrow-derived macrophages (BMDMs). Instead, addition of Rg6 to LPS-activated BMDMs augmented IL-10 expression, whereas it inhibited inflammatory signaling, such as by nuclear factor κB activation and mitogen-activated protein kinases. Furthermore, Rg6 significantly induced miR-146a, an operator miRNA for anti-inflammation, in BMDMs. Collectively, these data indicate that Rg6 inhibits inflammatory responses through the induction of IL-10 and miR-146a.

SIRT3 promotes antimycobacterial defenses by coordinating mitochondrial and autophagic functions.

SIRT3 (sirtuin 3), a mitochondrial protein deacetylase, maintains respiratory function, but its role in the regulation of innate immune defense is largely unknown. Herein, we show that SIRT3 coordinates mitochondrial function and macroautophagy/autophagy activation to promote anti-mycobacterial responses through PPARA (peroxisome proliferator activated receptor alpha). SIRT3 deficiency enhanced inflammatory responses and mitochondrial dysfunction, leading to defective host defense and pathological inflammation during mycobacterial infection. Antibody-mediated depletion of polymorphonuclear neutrophils significantly increased protection against mycobacterial infection in sirt3-/- mice. In addition, mitochondrial oxidative stress promoted excessive inflammation induced by Mycobacterium tuberculosis infection in sirt3-/- macrophages. Notably, SIRT3 was essential for the enhancement of PPARA, a key regulator of mitochondrial homeostasis and autophagy activation in the context of infection. Importantly, overexpression of either PPARA or TFEB (transcription factor EB) in sirt3-/- macrophages recovered antimicrobial activity through autophagy activation. Furthermore, pharmacological activation of SIRT3 enhanced antibacterial autophagy and functional mitochondrial pools during mycobacterial infection. Finally, the levels of SIRT3 and PPARA were downregulated and inversely correlated with TNF (tumor necrosis factor) levels in peripheral blood mononuclear cells from tuberculosis patients. Collectively, these data demonstrate a previously unappreciated function of SIRT3 in orchestrating mitochondrial and autophagic functions to promote antimycobacterial responses. Abbreviations: Ab: antibody; BCG: M. bovis Bacillus Calmette-Guérin; Baf-A1: bafilomycin A1; BMDMs: bone marrow-derived macrophages; CFU: colony forming unit; CXCL5: C-X-C motif chemokine ligand 5; EGFP: enhanced green fluorescent protein; ERFP: enhanced red fluorescent protein; FOXO3: forkhead box O3; HC: healthy controls; H&E: haematoxylin and eosin; HKL: honokiol; IHC: immunohistochemistry; IL1B: interleukin 1 beta; IL6: interleukin 6; IL12B: interleukin 12B; MDMs: monocyte-derived macrophages; MMP: mitochondrial membrane potential; Mtb: Mycobacterium tuberculosis; PBMC: peripheral blood mononuclear cells; PBS: phosphate buffered saline; PMN: polymorphonuclear neutrophil; PPARA: peroxisome proliferator activated receptor alpha; ROS: reactive oxygen species; SIRT3: sirtuin 3; TB: tuberculosis; TEM: transmission electron microscopy; TFEB: transcription factor EB; TNF: tumor necrosis factor.

Rufomycin Targets ClpC1 Proteolysis in Mycobacterium tuberculosis and M. abscessus.

ClpC1 is an emerging new target for the treatment of Mycobacterium tuberculosis infections, and several cyclic peptides (ecumicin, cyclomarin A, and lassomycin) are known to act on this target. This study identified another group of peptides, the rufomycins (RUFs), as bactericidal to M. tuberculosis through the inhibition of ClpC1 and subsequent modulation of protein degradation of intracellular proteins. Rufomycin I (RUFI) was found to be a potent and selective lead compound for both M. tuberculosis (MIC, 0.02 µM) and Mycobacterium abscessus (MIC, 0.4 µM). Spontaneously generated mutants resistant to RUFI involved seven unique single nucleotide polymorphism (SNP) mutations at three distinct codons within the N-terminal domain of clpC1 (V13, H77, and F80). RUFI also significantly decreased the proteolytic capabilities of the ClpC1/P1/P2 complex to degrade casein, while having no significant effect on the ATPase activity of ClpC1. This represents a marked difference from ecumicin, which inhibits ClpC1 proteolysis but stimulates the ATPase activity, thereby providing evidence that although these peptides share ClpC1 as a macromolecular target, their downstream effects are distinct, likely due to differences in binding.

Activity of LCB01-0371, a Novel Oxazolidinone, against Mycobacterium abscessus.

Mycobacterium abscessus is a highly pathogenic drug-resistant rapidly growing mycobacterium. In this study, we evaluated the in vitro, intracellular, and in vivo activities of LCB01-0371, a novel and safe oxazolidinone derivative, for the treatment of M. abscessus infection and compared its resistance to that of other oxazolidinone drugs. LCB01-0371 was effective against several M. abscessus strains in vitro and in a macrophage model of infection. In the murine model, a similar efficacy to linezolid was achieved, especially in the lungs. We induced laboratory-generated resistance to LCB01-0371; sequencing analysis revealed mutations in rplC of T424C and G419A and a nucleotide insertion at the 503 position. Furthermore, LCB01-0371 inhibited the growth of amikacin-, cefoxitin-, and clarithromycin-resistant strains. Collectively, our data indicate that LCB01-0371 might represent a promising new class of oxazolidinones with improved safety, which may replace linezolid for the treatment of M. abscessus.

Ohmyungsamycins promote antimicrobial responses through autophagy activation via AMP-activated protein kinase pathway.

The induction of host cell autophagy by various autophagy inducers contributes to the antimicrobial host defense against Mycobacterium tuberculosis (Mtb), a major pathogenic strain that causes human tuberculosis. In this study, we present a role for the newly identified cyclic peptides ohmyungsamycins (OMS) A and B in the antimicrobial responses against Mtb infections by activating autophagy in murine bone marrow-derived macrophages (BMDMs). OMS robustly activated autophagy, which was essentially required for the colocalization of LC3 autophagosomes with bacterial phagosomes and antimicrobial responses against Mtb in BMDMs. Using a Drosophila melanogaster-Mycobacterium marinum infection model, we showed that OMS-A-induced autophagy contributed to the increased survival of infected flies and the limitation of bacterial load. We further showed that OMS triggered AMP-activated protein kinase (AMPK) activation, which was required for OMS-mediated phagosome maturation and antimicrobial responses against Mtb. Moreover, treating BMDMs with OMS led to dose-dependent inhibition of macrophage inflammatory responses, which was also dependent on AMPK activation. Collectively, these data show that OMS is a promising candidate for new anti-mycobacterial therapeutics by activating antibacterial autophagy via AMPK-dependent signaling and suppressing excessive inflammation during Mtb infections.

Ionizing Radiation Induces Innate Immune Responses in Macrophages by Generation of Mitochondrial Reactive Oxygen Species.

During radiotherapy for tumors, the innate immune system also responds to ionizing radiation and induces immune modulation. However, little is known about the molecular mechanisms by which radiation modulates innate immune responses. In this study, we observed that radiation triggered the generation of mitochondrial reactive oxygen species (mROS), leading to innate immune responses in murine bone marrow-derived macrophages (BMDM). Radiation-induced mROS was essential for robust induction of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-12p40 mRNA and protein in BMDM. Exposure to radiation also led to rapid activation of the mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-κB pathways in BMDM. Notably, radiation-induced MAPK activation and NF-κB signaling were regulated by mROS in macrophages. Additionally, radiation-induced expression of TNF-α, IL-6 and IL-12p40 was dependent on JNK, p38 and NF-κB activation in BMDM. These data suggest a key role for radiation-induced pro-inflammatory responses and activation of the MAPK and NF-κB pathways through a triggering mechanism involving mROS generation.

Mycobacterium abscessus ESX-3 plays an important role in host inflammatory and pathological responses during infection.

Mycobacterial ESX systems are often related to pathogenesis during infection. However, little is known about the function of ESX systems of Mycobacterium abscessus (Mab). This study focuses on the Mab ESX-3 cluster, which contains major genes such as esxH (Rv0288, low molecular weight protein antigen 7; CFP-7) and esxG (Rv0287, ESAT-6 like protein). An esx-3 (MAB 2224c-2234c)-deletional mutant of Mab (Δesx) was constructed and used to infect murine and human macrophages. We then investigated whether Mab Δesx modulated innate host immune responses in macrophages. Mab Δesx infection resulted in less pathological and inflammatory responses. Additionally, Δesx resulted in significantly decreased activation of inflammatory signaling and cytokine production in macrophages compared to WT. Moreover, recombinant EsxG·EsxH (rEsxGH) proteins encoded by the ESX-3 region showed synergistic enhancement of inflammatory cytokine generation in macrophages infected with Δesx. Taken together, our data suggest that Mab ESX-3 plays an important role in inflammatory and pathological responses during Mab infection.

Is Varicocelectomy Useful for Subfertile Men with Isolated Teratozoospermia?

OBJECTIVE: To evaluate the effect of microsurgical varicocelectomy on sperm morphology in subfertile men with isolated teratozoospermia and a clinically palpable varicocele. MATERIALS AND METHODS: A retrospective review was performed of men with isolated teratozoospermia who had undergone microsurgical varicocelectomy. Semen analyses following varicocelectomy were performed at 2- to 3-month intervals. Responders to the varicocelectomy were defined as those who fulfilled the following criteria in semen analysis: (1) an improvement in percentage of normal forms to ≥4% and (2) normal results for all other semen parameters. RESULTS: A total of 80 patients underwent unilateral varicocelectomy (n = 49) or bilateral varicocelectomy (n = 31) (mean age of 36 ± 0.4 years, range 24-44). After a mean postoperative follow-up period of 6.8 months, the mean percentage of normal sperm forms increased from 0.9% to 3.5% (P < .001), and the mean sperm motility improved from 50.5% to 56.6% (P = .004). The number of responders to the surgery was 16 (20%), and their mean percentage of normal forms increased from 1.3% to 9.1%. The mean preoperative and postoperative semen volume and sperm concentration were not different (P > .05). No differences were observed in age, current smoking status, body mass index, or grade of varicocele between responders and nonresponders. CONCLUSION: Varicocelectomy might be an option for treating subfertile patients with isolated teratozoospermia and a clinically apparent varicocele. However, patients should be advised that the surgery helps in limited patients only because more patients did not benefit from the surgery.

Success rate of microsurgical multiple testicular sperm extraction and sperm presence in the ejaculate in korean men with y chromosome microdeletions.

PURPOSE: We assessed the frequency of azoospermia factor a (AZFa), AZFb, and AZFc deletions and examined correlations between the deletion sites and the success rates of sperm presence within the ejaculate and surgical sperm retrieval in Korean men. MATERIALS AND METHODS: A total of 1,919 azoospermic and severely oligozoospermic men were assessed for Y chromosome microdeletions. Among them, 168 men with AZF deletions were identified and their medical records were reviewed. RESULTS: Of the total 168 men with AZF deletions, there were 13 with AZFa, 10 with AZFb, 95 with AZFc, 37 with AZFbc, and 13 with AZFabc deletions. Of the 95 men with isolated AZFc deletion, 51 had the presence of sperm in the ejaculate. Of the infertile men with any other deletion, however, only two patients (one man with AZFb deletion and another with AZFbc deletion) showed the presence of sperm in the ejaculate. The success rates for surgical sperm retrieval were 7.1% (1/14) in men with AZFbc deletion and 54.8% (17/31) in the isolated AZFc deletion group. No sperm was obtained from the patients with AZFa or AZFb deletions who underwent microsurgical sperm retrieval. In the isolated AZFc deletion group, there were significant differences between azoospermic and severely oligozoospermic patients in terms of testicular volume and serum levels of follicle-stimulating hormone and luteinizing hormone, whereas no significant differences were found when the group was divided by surgical sperm retrieval outcomes. CONCLUSIONS: Deletions of the AZFa and AZFb regions are associated with severe spermatogenetic impairment. However, more than half of men with an AZFc deletion had sperm within the ejaculate or testis for in vitro fertilization with intracytoplasmic sperm injection.

Concordance of Ureaplasma urealyticum and Mycoplasma hominis in infertile couples: impact on semen parameters.

OBJECTIVE: To assess the relationship between mycoplasma infection and human infertility, we determined the concordance of Ureaplasma urealyticum (UU) and Mycoplasma hominis (MH) detection in infertile and fertile couples, and assessed semen parameters in both groups. METHODS: Fifty infertile couples without a female factor attending a fertility clinic and 48 fertile couples were randomly screened for UU and MH. The concordance between partners was compared between the fertile and infertile groups. Semen and endocervical specimens were evaluated using the commercially available Mycofast Evolution2 test. RESULTS: UU was detected in 24 semen specimens (48%) from the infertile men, in 12 specimens from fertile men (25%), in 20 endocervical specimens from infertile women (40%), and 11 from fertile women (22.9%). UU was detected higher in infertile men than in fertile men (P = .022). The concordance of UU was higher in infertile couples (32%) than in fertile couples (12.5%, P = .022). The concordance of MH between male and female partners in the 2 groups did not differ significantly. The mean values of total motility, progressive motility, normal morphology, vitality, and total motile sperm count were significantly lower in sperm from infertile men than from fertile men. Progressive motility and vitality were significantly lower in UU-positive men than in men without UU, and low total motility and total motile sperm count were significantly related to the presence of MH. CONCLUSION: Clinicians should consider the roles of UU and MH in infertility and routinely screen infertile couples for the presence of these mycoplasma species.

Urologic complications following obstetric and gynecologic surgery.

PURPOSE: Urologic injuries occur frequently during surgery in the pelvic cavity. Inadequate diagnosis and treatment may lead to severe complications and side effects. This investigation examined the clinical features of urologic complications following obstetric and gynecologic surgery. MATERIALS AND METHODS: We accumulated 47,318 obstetric and gynecologic surgery cases from 2007 to 2011. Ninety-seven patients with urological complications were enrolled. This study assessed the causative disease and surgical approach, type, and treatment method of the urologic injury. RESULTS: Of these 97 patients, 69 had bladder injury, 23 had ureteral injury, 2 had vesicovaginal fistula, 2 had ureterovaginal fistula, and 1 had renal injury. With respect to injury rate by specific surgery, laparoscopic-assisted radical vaginal hysterectomy was the highest with 3 of 98 cases, followed by radical abdominal hysterectomy with 15 of 539 cases. All 69 cases of bladder injury underwent primary suturing during surgery without complications. Of 14 cases with an early diagnosis of ureteral injury, 7 had a ureteral catheter inserted, 5 underwent ureteroureterostomy, and 2 underwent ureteroneocystostomy. Of nine cases with a delayed diagnosis of ureteral injury, ureteral catheter insertion was carried out in three cases, four cases underwent ureteroureterostomy, and two cases underwent ureteroneocystostomy. CONCLUSIONS: Bladder injury was the most common urological injury during obstetric and gynecologic surgery, followed by ureteral injury. The variety of injured states, difficulty of diagnosis, and time to complete cure were much greater among patients with ureteral injuries. Early diagnosis and urologic intervention is important for better outcomes.

Causes and classification of male infertility in Korea.

OBJECTIVE: The aim of this study is to investigate the various causes of male infertility using multiple approaches. METHODS: Nine-hundred-twenty infertile male patients were analyzed at their first visit with one physician between January 1 and December 31, 2009. All patients were subjected to physical examination, semen analysis and azoospermic patients underwent hormonal testing, chromosomal tests, and testicular biopsy. Semen analysis was based on the definition of the World Health Organization. RESULTS: Among the 920 patients, 555 patients (60.3%) had semen results within the normal range, 269 patients (29.2%) within the abnormal range, and 96 (10.5%) were diagnosed with azoospermia. Varicoceles were diagnosed in 84 of the 555 normal-range patients (15.1%) and in 113 of the 269 abnormal-range patients (42.0%). Of the 96 patients with azoospermia, 24 patients (25%) were diagnosed with obstructive azoospermia, 68 patients (71%) with non-obstructive azoospermia, and 4 patients (4%) with retrograde ejaculation. CONCLUSION: Various causes of male infertility have been reported and diverse treatment methods can be adopted for each cause. In this regard, research must be conducted on a larger number of patients to accurately assess the various causes of infertility in Korean patients and to investigate various infertility treatment methods.

Prevalence of painful bladder syndrome/interstitial cystitis-like symptoms in women: a population-based study in Korea.

OBJECTIVE: Despite growing clinical interest in painful bladder syndrome/interstitial cystitis (PBS/IC, also known as bladder pain syndrome), estimating its prevalence is difficult because of its variable presentation and the lack of clear diagnostic criteria. In this study, we estimated the prevalence of PBS/IC-like urinary symptoms in adult women in the general population of South Korea. METHODS: A population-based cross-sectional telephone survey was conducted among 2,323 women (18-71 years of age), selected by geographically stratified random sampling, based on Korean census data. The survey was performed by trained interviewers between September 22, 2008, and October 6, 2008. All participants were interviewed by telephone using a validated questionnaire, the O'Leary-Sant IC Symptom and Problem (OLS) index. Women with high symptom and problem index scores of 12 or greater and scores of two or greater for pain and nocturia symptoms were considered to have "probable PBS/IC," according to previously suggested criteria. RESULTS: After exclusions, a total of 2,300 respondents were included. The severity of symptoms increased with age. Eight respondents (0.35%) reported severe symptoms and problems (OLS survey scores of ≥12). Of these, six (261/100,000 or 0.26%, 95% CI 242-278) met previously suggested criteria for probable PBS/IC. CONCLUSION: The prevalence of PBS/IC-like urinary symptoms in South Korean women appeared to be lower than in Europe and the United States, and similar to that of Japan, according to common criteria. Screening for symptoms that are consistent with the disease may improve our understanding of its true prevalence.

One-Year Surgical Outcomes and Quality of Life after Minimally Invasive Sling Procedures for the Treatment of Female Stress Urinary Incontinence: TVT SECUR(R) vs. CureMesh(R).

PURPOSE: We compared the efficacy and safety of two minimally invasive sling procedures used to treat female stress urinary incontinence (SUI), tension-free vaginal tape (TVT) SECUR(R) and CureMesh(R), and assessed the 1-year surgical outcomes. MATERIALS AND METHODS: Sixty women with SUI were assigned to undergo either the TVT SECUR (n=38) or CureMesh (n=22) procedures between April 2007 and June 2008. Patients were monitored via outpatient visits at 1 month, 3 months, and 1 year after surgery. The efficacy of these procedures was evaluated by the cough test or by a urodynamic study. At these postoperative visits, the patients also completed several questionnaires, including incontinence quality of life, patient's perception of urgency severity, the scored form of the Bristol Female Lower Urinary Tract Symptoms, visual analog scale, and questions about perceived benefit, satisfaction, and willingness to undergo the same operation again. The objective cure rate was defined as no leakage during the cough test with a full bladder. The subjective cure rate was evaluated by self-assessment of goal achievement performed 1 year postoperatively. RESULTS: The two groups were similar in preoperative characteristics and urodynamic parameters. The objective cure rates were similar between TVT SECUR and CureMesh (68.4% vs. 77.3%). All respondents reported improvement after surgery. There were no intra-operative complications. CONCLUSIONS: Our results showed that the TVT SECUR and CureMesh procedures are both safe and simple to perform and have no significant differences in efficacy. Comparative studies with long-term follow-up are warranted to determine the true efficacy of these procedures.

Efficacy of lidocaine spray as topical anesthesia for outpatient rigid cystoscopy in women: a prospective, randomized, double-blind trial.

OBJECTIVES: To determine the efficacy and feasibility of urethral lidocaine spray for rigid cystoscopy in female outpatients, compared with lidocaine gel for patient tolerance of pain. METHODS: A total of 144 consecutive women scheduled to undergo rigid cystoscopy were randomized to receive either 10 mL of lidocaine gel (group 1, n = 48), five metered doses of 10% lidocaine spray (group 2, n = 48), or 10 mL of plain lubricating gel (group 3, n = 48). Patients recorded their level of pain perception during gel instillation or spray and immediately after the procedure using the visual analogue scale (VAS) and a 5-point verbal descriptor scale. RESULTS: The perception of pain was significantly decreased when lidocaine spray was used (mean VAS scores 1.9 +/- 1.3 for group 2 versus 3.9 +/- 2.2 for group 3; P <0.001). There was no difference in pain perception when lidocaine spray or gel was used (P = 0.113). Four patients in group 1 and 1 in group 2 but 10 in group 3 requested post-cystoscopic analgesics. There was a slight increase in pain perception on the VAS during the intraurethral application with lidocaine spray (2.2 +/- 1.5) compared with lidocaine gel (0.7 +/- 1.1) or plain gel (1.0 +/- 1.4). CONCLUSIONS: Lidocaine spray and lidocaine gel produce similar anesthetic effects in women undergoing rigid cystoscopy. However, concerns about the mild discomfort associated with delivery of the spray need to be resolved through future technical improvement before it gains widespread use.

The impact of tension-free vaginal tape on overactive bladder symptoms in women with stress urinary incontinence: significance of detrusor overactivity.

PURPOSE: We assessed the impact of the tension-free vaginal tape procedure on overactive bladder in women with stress urinary incontinence to determine the change in preoperative overactive bladder symptoms and the significance of detrusor overactivity in postoperative outcomes. MATERIALS AND METHODS: From January 2003 to December 2004 a total of 549 consecutive women with stress urinary incontinence underwent the tension-free vaginal tape procedure. Of these patients 180 (32.8%) had overactive bladder syndrome. Finally 132 women (mean age 53.2 years) with followup of at least 3 months were included in analysis. Patients with the overactive bladder syndrome with or without detrusor overactivity based on urodynamic study were subdivided into 94 (71.2%) patients with the overactive bladder syndrome without detrusor overactivity and 38 (28.8%) with the overactive bladder syndrome with detrusor overactivity. RESULTS: The complete resolution of all overactive bladder symptoms based on 3-day frequency volume chart and a validated questionnaire was demonstrated in 23.5% (31 of 132 patients). The detrusor overactivity present group showed significantly greater urinary leakage per 1-hour pad test, maximal detrusor pressure and detrusor pressure at maximal flow rate than the detrusor overactivity absent group. There was no significant difference in the 2 groups in terms of cure rate for stress urinary incontinence. The detrusor overactivity present group had a significantly higher resolution rate than the detrusor overactivity absent group for overactive bladder (36.8 vs 18.1 %, p = 0.021). CONCLUSIONS: The tension-free vaginal tape procedure can be performed in women with stress urinary incontinence and overactive bladder including urge incontinence even if the patient has detrusor overactivity on urodynamic study. However, patients should be fully advised of the possibility of persistent overactive bladder symptoms and treatment for those symptoms after tension-free vaginal tape should be considered.

Routine screening for classical azoospermia factor deletions of the Y chromosome in azoospermic patients with Klinefelter syndrome.

AIM: To evaluate the occurrence of classical azoospermia factor (AZF) deletions of the Y chromosome as a routine examination in azoospermic subjects with Klinefelter syndrome (KS). METHODS: Blood samples were collected from 95 azoospermic subjects with KS (91 subjects had a 47,XXY karyotype and four subjects had a mosaic 47,XXY/46,XY karyotype) and a control group of 93 fertile men. The values of testosterone, follicle stimulating hormone (FSH) and luteinizing hormone (LH) were measured. To determine the presence of Y chromosome microdeletions, polymerase chain reaction (PCR) of five sequence-tagged site primers (sY84, sY129, sY134, sY254, sY255) spanning the AZF region, was performed on isolated genomic DNA. RESULTS: Y chromosome microdeletions were not found in any of the 95 azoospermic subjects with KS. In addition, using similar conditions of PCR, no microdeletions were observed in the 93 fertile men evaluated. The level of FSH in KS subjects was higher than that in fertile men (38.2 +/- 10.3 mIU/mL vs. 5.4 +/- 2.9 mIU/mL, P < 0.001) and the testosterone level was lower than that in the control group (1.7 +/- 0.3 ng/mL vs. 4.3 +/- 1.3 ng/mL, P < 0.001). CONCLUSION: Our data and review of the published literature suggest that classical AZF deletions might not play a role in predisposing genetic background for the phenotype of azoospermic KS subjects with a 47,XXY karyotype. In addition, routine screening for the classical AZF deletions might not be required for these subjects. Further studies including partial AZFc deletions (e.g. gr/gr or b2/b3) are necessary to establish other mechanism underlying severe spermatogenesis impairment in KS.

Urodynamic studies in women with stress urinary incontinence: Significant bacteriuria and risk factors.

AIM: A prospective study was performed to determine the incidence of significant bacteriuria and to identify the risk factors for bacteriuria after urodynamic studies (UDSs) in women with urodynamic stress urinary incontinence (SUI). METHODS: A total of 225 women with urodynamic SUI were evaluated. All women were negative on double-screened urine cultures, in clean-catch midstream urine (MSU) specimens, before UDS. Another urine specimen was obtained for urinalysis and culture at 3-7 days after UDS. Urinary culture with 10(5) CFU/ml or more was regarded as significant bacteriuria. To identify the risk factors for significant bacteriuria, the clinical characteristics of all patients including age, BMI, parity, medical and operation history, degree of pelvic organ prolapse, results of urinalysis, and UDS were evaluated. RESULTS: The prevalence of significant bacteriuria was 6.2%. The most common identified microorganism was Escherichia coli (57.1%). Univariate analysis demonstrated that a history of recurrent urinary tract infection (UTI; P = 0.002) and urological surgery or procedure (P = 0.02) were significant predictors of significant bacteriuria. On multiple logistic regression analysis the past history of recurrent UTI was the only significant independent risk factor (OR = 28.5, 95% CI = 4.309-188.488, P = 0.009). CONCLUSIONS: This study suggests that for most women with SUI it may be unnecessary to use preventive prophylactic antibiotics in UDS. However, our results suggest that in patients with a previous history of recurrent UTI or urologic surgery the risk for significant bacteriuria is increased and use of prophylactic antibiotics should be considered.

Accuracy and precision of a new portable ultrasound scanner, the BME-150A, in residual urine volume measurement: a comparison with the BladderScan BVI 3000.

The objective of the study was to determine the relative accuracy of a new portable ultrasound unit, BME-150A, and the BladderScan BVI 3000, as assessed in comparison with the catheterized residual urine volume. We used both of these machines to prospectively measure the residual urine volumes of 89 patients (40 men and 49 women) who were undergoing urodynamic studies. The ultrasound measurements were compared with the post-scan bladder volumes obtained by catheterization in the same patients. The ultrasounds were followed immediately (within 5 min) by in-and-out catheterizations while the patients were in a supine position. There were a total of 116 paired measurements made. The BME-150A and the BVI 3000 demonstrated a correlation with the residual volume of 0.92 and 0.94, and a mean difference from the true residual volume of 7.8 and 3.6 ml, respectively. Intraclass correlation coefficients for the accuracy of the two bladder scans were 0.90 for BME-150A and 0.95 for BVI 3000. The difference of accuracy between the two models was not significant (p = 0.2421). There were six cases in which a follow-up evaluation of falsely elevated post-void residual urine volume measurements on the ultrasound studies resulted in comparatively low catheterized volumes, with a range of differences from 66 to 275.5 ml. These cases were diagnosed with an ovarian cyst, uterine myoma, or uterine adenomyosis on pelvic ultrasonography. The accuracy of the BME-150A is comparable to that of the BVI 3000 in estimating the true residual urine volumes and is sufficient enough for us to recommend its use as an alternative to catheterization.