The Engineered Lysin, CF-370, is Active Against Antibiotic-Resistant Gram-Negative Pathogens In vitro and Synergizes with Meropenem in Experimental Pseudomonas aeruginosa Pneumonia.

BACKGROUND: Lysins (cell wall hydrolases) targeting Gram-negative organisms require engineering to permeabilize the outer membrane and access subjacent peptidoglycan to facilitate killing. In the current study, the potential clinical utility for engineered lysin, CF-370, was examined in vitro and in vivo against Gram-negative pathogens important in human infections. METHODS: MICs and bactericidal activity were determined using standard methods. An in vivo proof-of-concept efficacy study was conducted using a rabbit acute pneumonia model caused by Pseudomonas aeruginosa. RESULTS: CF-370 exhibited potent antimicrobial activity, with MIC50/90 values (in µg/mL) for: P. aeruginosa, 1/2; Acinetobacter baumannii, 1/1; Escherichia coli, 0.25/1; Klebsiella pneumoniae, 2/4; Enterobacter cloacae 1/4; and Stenotrophomonas maltophilia 2/8. CF-370 furthermore demonstrated: i) bactericidal activity; (ii) activity in serum; iii) a low propensity for resistance; iv) anti-biofilm activity; and v) synergy with antibiotics. In the pneumonia model, CF-370 alone decreased bacterial densities in lungs, kidneys and spleen vs. vehicle control, and demonstrated significantly increased efficacy when combined with meropenem (vs either agent alone). CONCLUSIONS: CF-370 is the first engineered lysin described with potent broad spectrum in vitro activity against multiple clinically-relevant Gram-negative pathogens, as well as potent in vivo efficacy in an animal model of severe invasive multi-system infection.

The Incidence of Pelvic and Low Back Pain in Patients with Pelvic Organ Prolapse.

INTRODUCTION AND HYPOTHESIS: To define the prevalence and incidence of pelvic/low back pain in patients with pelvic organ prolapse (POP). METHODS: Patients presenting for POP to three urogynecology centers in the US, UK, and Chile were enrolled in an IRB-approved cross-sectional study assessing pain, GU, GI and sexual function symptoms. For prevalence, symptoms were noted as present if the participant recorded the symptom and reported the degree of bother as "somewhat," "a moderate amount," or "a lot." For incidence, participants were queried if the symptom's onset concurred with the POP. We also queried if they perceived the symptom was worsened by their POP. RESULTS: Two hundred five participants were recruited: 100 from the US, 46 from the UK, and 59 from Chile. One US participant was excluded due a missing examination. The prevalence of pelvic pain was 42%. Seventy-three percent of these participants reported the onset of pelvic pain coinciding with prolapse onset, and 81% endorsed worsening pelvic pain with POP. The prevalence of low back pain was 46%, with 30% reporting the onset coincided with the onset of POP and 44% responded that prolapse worsened their pain. CONCLUSION: A higher proportion of participants than expected endorsed pelvic/low back pain. Among patients with pelvic pain, the majority experienced symptom onset with POP onset and a worsening of pain with POP. While roughly half of participants reported low back pain; a minority correlated this to their POP. These findings highlight a high incidence of pelvic pain, challenging the perception of POP as a painless condition.

The effect of preoperative phenazopyridine on short-term urinary retention following urogynecologic surgery.

INTRODUCTION AND HYPOTHESIS: Previous studies have found that administration of phenazopyridine decreased short-term urinary retention following surgery but other more recent trials have shown mixed results. This study sought to investigate the potential benefit of preoperative administration of oral phenazopyridine in relation to the prevention of short-term urinary retention following urogynecologic surgery. METHODS: This is a retrospective cohort study of a convenience sample of women undergoing urogynecologic surgery from June 2016 to March 2019. Following surgery, subjects underwent a standardized retrograde voiding trial. The data had previously been gathered from a prior prospective trial at our institution (Kesty et al. Int Urogynecol J 31(9):1899-1905, 11). Chart review was performed to determine whether patients that received 200 mg of preoperative oral phenazopyridine to better visualize ureteral efflux during cystourethroscopy were more or less likely to pass their postoperative voiding trial. Bivariate statistical analysis was performed as well as a multivariate logistic regression model. RESULTS: A total of 165 subjects were included in the final analysis; 100 who did not receive preoperative phenazopyridine and 65 who did receive phenazopyridine. There was no statistical difference between voiding trial pass rates following urogynecologic surgery between those who did not receive preoperative phenazopyridine compared to those who did [77% (77/100) and 82% (53/65), respectively, p = 0.37)]. The multivariate logistic regression model demonstrated no difference in postoperative voiding trial pass rates among those who received preoperative phenazopyridine compared to those who did not (OR 1.7, 95% CI: 0.53, 5.8). CONCLUSIONS: Preoperative administration of oral phenazopyridine does not decrease short-term urinary retention following urogynecologic surgery.

A Cost-Effectiveness Analysis of Post-Void Residual Bladder Scan Thresholds in the Postoperative Setting.

INTRODUCTION AND HYPOTHESIS: To identify the optimal cost-effectiveness threshold of post-void residual (PVR) by bladder scan in postoperative urogynecologic patients. METHODS: A cost-effectiveness analysis was performed as a secondary analysis of a previously published study of patients undergoing urogynecologic procedures with planned voiding trials, setting thresholds for postoperative PVR bladder scan volumes at 100 ml, 150 ml, and 200 ml. Patient-based scenarios were modeled for ambulatory office or emergency department (ED) resource utilization and to determine the cost-effectiveness of each threshold. Costs were obtained from a southeastern academic medical center, only utilizing direct medical costs and hospital costs, not including societal costs. Quality-adjusted life years (QALY's) were used as health outcomes determining the incremental cost-effectiveness ratio (ICER). RESULTS: A total of 151 patients from the original study were included. A willingness to pay threshold of $100,000 per QALY was assumed. A PVR of 100 ml exceeded this at $373,824. A PVR threshold of 150 ml was dominant (-$1,211,716), while minimizing ED visits for postoperative urinary retention (POUR) and unnecessary clinic appointments. While a PVR of 200 ml appeared a cost-effective strategy (-$488,389), there was increased ED utilization and under-detection of postoperative urinary retention (POUR). CONCLUSION: A PVR threshold of 100 ml created a healthcare system burden due to increased office voiding trials. Both PVR thresholds of 150 ml and 200 ml were cost-effective strategies; however, ED utilization for POUR increased with 200 ml. Utilizing 150 ml as the PVR cut-off proved the most cost-effective strategy, avoiding POUR under-detection and undue health costs.

Exebacase Is Active In Vitro in Pulmonary Surfactant and Is Efficacious Alone and Synergistic with Daptomycin in a Mouse Model of Lethal Staphylococcus aureus Lung Infection.

Exebacase (CF-301) is a novel antistaphylococcal lysin (cell wall hydrolase) in phase 3 of clinical development for the treatment of Staphylococcus aureus bacteremia, including right-sided endocarditis, used in addition to standard-of-care antibiotics. In the current study, the potential for exebacase to treat S. aureus pneumonia was explored in vitro using bovine pulmonary surfactant (Survanta) and in vivo using a lethal murine pneumonia model. Exebacase was active against a set of methicillin-sensitive S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA) strains, with an MIC90 of 2 µg/ml (n = 18 strains), in the presence of a surfactant concentration (7.5%) inhibitory to the antistaphylococcal antibiotic daptomycin, which is inactive in pulmonary environments due to specific inhibition by surfactant. In a rigorous test of the ability of exebacase to synergize with antistaphylococcal antibiotics, exebacase synergized with daptomycin in the presence of surfactant in vitro, resulting in daptomycin MIC reductions of up to 64-fold against 9 MRSA and 9 MSSA strains. Exebacase was also observed to facilitate the binding of daptomycin to S. aureus and the elimination of biofilm-like structures formed in the presence of surfactant. Exebacase (5 mg/kg of body weight 1 time every 24 h [q24h], administered intravenously for 3 days) was efficacious in a murine model of staphylococcal pneumonia, resulting in 50% survival, compared to 0% survival with the vehicle control; exebacase in addition to daptomycin (50 mg/kg q24h for 3 days) resulted in 70% survival, compared to 0% survival in the daptomycin-alone control group. Overall, exebacase is active in pulmonary environments and may be appropriate for development as a treatment for staphylococcal pneumonia.

Mental health among clinicians: what do we know and what can we do?

Mental health and mental health disorders among clinicians remain a taboo, despite increasing evidence showing the direct impact on medical teams and patient care. This editorial is aimed at increasing awareness of mental issues amongst healthcare professionals, identifying perceived barriers to seeking help, and suggesting ways in which to seek help. Mental health disorders, including anxiety and depression, are prevalent from medical school, leading to increased burnout and suicide risks at later stages of a clinician's career. There is often a reluctance to seek help, particularly amongst the surgical specialties, caused by self-criticism, lack of convenient access and the potential negative impact on medical licensure. This editorial has been written in loving memory of our colleague, friend and board member Dr. Nikolaus Veit-Rubin, who sadly passed away at the beginning of the year. It is written in the hope of highlighting the importance of maintaining mental wellbeing amongst the medical team, supporting help-seeking behaviour and changing attitudes toward mental health disorders amongst clinicians.

Treatment outcomes of overactive bladder with combined therapies of botulinum toxin injections and oral agents.

OBJECTIVE: Treatment options for refractory overactive bladder (OAB) following behavioral modifications and oral OAB medications remain limited. Up to 33% of women fail botulinum toxin injections (Amundsen et al. Eur Urol. 74(1):66-73, 7). This study evaluates the effectiveness of combining oral OAB agents with botulinum toxin in subjects who failed botulinum toxin therapy alone. METHODS: This is a retrospective observational study. Eligible women were those who received botulinum toxin injections for OAB between 2013 and 2018 at one academic institution. Women were given the option of oral medications as add-back therapy following failed treatment with botulinum toxin alone. Treatment response was a subjective outcome, with subjects reporting being satisfied or unsatisfied. The primary outcome was the proportion of subjects who achieved satisfactory treatment response with the combination of oral OAB medications and botulinum toxin injections. A subanalysis was performed to further investigate any risk factors associated with poor response to combination treatment. Variables were analyzed using chi-squared or Fisher's exact test and Student's t-test or Mann-Whitney U when appropriate. RESULTS: A total of 107 charts were reviewed. Forty-five (48%) women failed botulinum toxin alone as a treatment; 26 (29%) elected to try one or more oral OAB medications. Of the 26 women who received the combination treatment, 17 (65%) reported satisfaction and 9 (35%) remained unsatisfied. Risk factors associated with treatment failure were not identified. CONCLUSION: This is an initial report on adding back oral OAB meds to patients who have failed botulinum toxin injections suggesting there may be a role for add-back oral OAB medications.

Factors affecting patient choice for continued observation versus intervention for pelvic organ prolapse.

INTRODUCTION AND HYPOTHESIS: To analyze the reasons for eventual choice of a therapeutic intervention in subjects who initially chose observation for bothersome pelvic organ prolapse (POP) over therapeutic intervention at their first urogynecology clinic visit. METHODS: This is a retrospective cohort study of women with bothersome POP who initially chose observation over therapeutic intervention at one institution from 2002 to 2015. Subjects were followed over time with sequential pelvic organ prolapse quantification examinations (POP-Q) and assessments of symptoms and bother utilizing non-validated standard questions. Subjects were divided into two groups: (1) those who chose continued observation versus (2) those who chose therapeutic intervention with either pessary or surgery. Demographic information, POP-Q examinations, and POP symptoms and bother were collected. We analyzed which clinical variables influenced patient decision to elect for therapeutic intervention. RESULTS: A total of 111 subjects were enrolled. The distribution of initial POP-Q stage was: stage 2 = 54%; stage 3 = 45%; stage 4 = 1%. Median follow-up was 24 months (range 6 and 110 months). At their last recorded visit, 73 subjects (66%) continued observation and 38 subjects (34%) chose pessary or surgical intervention. We investigated clinical factors for choosing intervention. Increase in POP symptom bother was the only variable that remained significant in determining patient choice of a therapeutic intervention (p < 0.001) after confounding factors were controlled for using multivariate regression analysis. CONCLUSION: In subjects with bothersome POP who initially choose observation and subsequently elect to pursue a therapeutic intervention, worsening symptom bother is the most important factor.

Novel Lytic Enzyme of Prophage Origin from Clostridium botulinum E3 Strain Alaska E43 with Bactericidal Activity against Clostridial Cells.

Clostridium botulinum is a Gram-positive, anaerobic, spore-forming bacterium capable of producing botulinum toxin and responsible for botulism of humans and animals. Phage-encoded enzymes called endolysins, which can lyse bacteria when exposed externally, have potential as agents to combat bacteria of the genus Clostridium. Bioinformatics analysis revealed in the genomes of several Clostridium species genes encoding putative N-acetylmuramoyl-l-alanine amidases with anti-clostridial potential. One such enzyme, designated as LysB (224-aa), from the prophage of C. botulinum E3 strain Alaska E43 was chosen for further analysis. The recombinant 27,726 Da protein was expressed and purified from E. coli Tuner(DE3) with a yield of 37.5 mg per 1 L of cell culture. Size-exclusion chromatography and analytical ultracentrifugation experiments showed that the protein is dimeric in solution. Bioinformatics analysis and results of site-directed mutagenesis studies imply that five residues, namely H25, Y54, H126, S132, and C134, form the catalytic center of the enzyme. Twelve other residues, namely M13, H43, N47, G48, W49, A50, L73, A75, H76, Q78, N81, and Y182, were predicted to be involved in anchoring the protein to the lipoteichoic acid, a significant component of the Gram-positive bacterial cell wall. The LysB enzyme demonstrated lytic activity against bacteria belonging to the genera Clostridium, Bacillus, Staphylococcus, and Deinococcus, but did not lyse Gram-negative bacteria. Optimal lytic activity of LysB occurred between pH 4.0 and 7.5 in the absence of NaCl. This work presents the first characterization of an endolysin derived from a C. botulinum Group II prophage, which can potentially be used to control this important pathogen.

Subjective versus objective determination of bladder emptying following urogynecological surgery: "do you feel that you completely emptied your bladder?"

INTRODUCTION AND HYPOTHESIS: We tested the hypothesis that women can subjectively determine if they have emptied their bladder after a spontaneous voiding attempt following urogynecological surgery to rule out post-operative urinary retention as determined by a voiding trial. METHODS: This is a prospective observational study of women undergoing urogynecological surgery at two academic institutions from June 2016 to March 2019. Following surgery, subjects underwent a voiding trial followed by measurement of the PVR via ultrasound bladder scan or straight catheterization. A successful voiding trial was defined as a PVR of ≤150 ml. Subjects were queried about their subjective sensation of bladder emptying; "Do you feel that you completely emptied your bladder?" Their responses were either "Yes", "No" or "I don't know". Their subjective responses were correlated with the voiding trial results using a Chi-squared analysis for sensitivity, specificity, and positive (PPV) and negative predictive values (NPV). RESULTS: A total of 266 subjects were included in the final evaluation. The screening subjective question had a sensitivity of 85.7% (CI 71.46 to 94.57%), a specificity of 91.5% (CI 87.01 to 94.79%), a PPV of 65.4% (CI 54.78 to 74.77%), and an NPV of 97.14% (CI 94.18 to 98.62%) to detect a failed voiding trial. CONCLUSIONS: The NPV of the subjective question regarding bladder emptying in the post-operative urogynecological setting is high at >97%, suggesting that it might be reasonable to forgo a formal voiding trial in patients who subjectively feel that they have emptied their bladder.

Optimized production of a biologically active Clostridium perfringens glycosyl hydrolase phage endolysin PlyCP41 in plants using virus-based systemic expression.

BACKGROUND: Clostridium perfringens, a gram-positive, anaerobic, rod-shaped bacterium, is the third leading cause of human foodborne bacterial disease and a cause of necrotic enteritis in poultry. It is controlled using antibiotics, widespread use of which may lead to development of drug-resistant bacteria. Bacteriophage-encoded endolysins that degrade peptidoglycans in the bacterial cell wall are potential replacements for antibiotics. Phage endolysins have been identified that exhibit antibacterial activities against several Clostridium strains. RESULTS: An Escherichia coli codon-optimized gene encoding the glycosyl hydrolase endolysin (PlyCP41) containing a polyhistidine tag was expressed in E. coli. In addition, The E. coli optimized endolysin gene was engineered for expression in plants (PlyCP41p) and a plant codon-optimized gene (PlyCP41pc), both containing a polyhistidine tag, were expressed in Nicotiana benthamiana plants using a potato virus X (PVX)-based transient expression vector. PlyCP41p accumulated to ~ 1% total soluble protein (100µg/gm f. wt. leaf tissue) without any obvious toxic effects on plant cells, and both the purified protein and plant sap containing the protein lysed C. perfringens strain Cp39 in a plate lysis assay. Optimal systemic expression of PlyCP41p was achieved at 2 weeks-post-infection. PlyCP41pc did not accumulate to higher levels than PlyCP41p in infected tissue. CONCLUSION: We demonstrated that functionally active bacteriophage PlyCP41 endolysin can be produced in systemically infected plant tissue with potential for use of crude plant sap as an effective antimicrobial agent against C. perfringens.

A narrative review of the epidemiology, diagnosis, and treatment of latent stress urinary incontinence.

AIMS: To review the current literature on the development of latent stress urinary incontinence (SUI) or SUI that occurs following pelvic organ prolapse therapy in patients who were continent prior to therapy. To make recommendations on how to best evaluate and manage patients with pelvic organ prolapse who are continent. METHODS: This is a narrative review using Google scholar and cross referencing identified manuscripts. We used the British Medical Journals grading of literature to assign letter grades to level of statements and recommendations. RESULTS: The literature suggest about 40% of women with advanced pelvic organ prolapse who are continent will develop SUI after surgery to correct their pelvic organ prolapse. Reduction cough stress testing can help identify those patients at risk to develop SUI with a PPV of 60%. Reduction cough stress testing is best done using large rectal swabs or scopettes. The literature on how to best manage patients with advanced pelvic organ prolapse and are continent to avoid latent SUI is mixed and the best recommendations are based on sound literature that is incomplete. CONCLUSIONS: Understanding that latent SUI is a potential complication in any subject with advanced pelvic organ prolapse is necessary to pre-operative counseling. Deciding on the best management strategy is not clear cut and therefore should involve patient input in the decision making process.

Defining normal apical vaginal support: a relook at the POSST study.

INTRODUCTION AND HYPOTHESIS: The purpose of this study is to reanalyze data from the original 2005 Pelvic Organ Support Study (POSST) data set to define normal values for apical Pelvic Organ Prolapse Quantification (POP-Q) points C and D and total vaginal length (TVL) in an asymptomatic population of women. METHODS: In this retrospective observational data-set review, patient were >18 years presenting for annual gynecologic exams to six centers in the United States. Data included demographics, questions about prolapse symptoms, and POP-Q points. Means and standard deviations were determined for each POP-Q point in the total population. RESULTS: The data set comprised 1011 women; 59 were excluded because they met criteria for having POP or were missing data. This left 948 for study. Mean age of our study population was 42 ± 14 years, and 45.6% were white, 25.1% black, and 25.2% Hispanic. One hundred fifty-six had a prior hysterectomy. Mean values with standard deviations (SD) for POP-Q values are as following: point C (vaginal cuff) -7.3 ± 1.5 cm, point C (cervix) -5.9 ± 1.5, point D -8.7 cm ± 1.5 cm, TVL (no hysterectomy) 9.8 cm ± 1.3 cm, and TVL (hysterectomy) 8.9 cm ± 1.5 cm. CONCLUSIONS: This data suggests normal values for POP-Q apical points in a population of patient with annual gynecological exams.

Transcultural translation and validation of the FIGO Assessment Scoring System (FASS) to Portuguese language.

INTRODUCTION AND HYPOTHESIS: Our aim was to validate the Brazilian Portuguese version of the International Federation of Gynecology and Obstetrics (FIGO) Assessment Scoring System (FASS) to identify and quantify signs and symptoms related to pelvic floor dysfunction. METHODS: One hundred and seventy-nine women aged 18-82 (47.68 ± 14.42) years participated in this validation study. Collected data included a sociodemographic and clinical questionnaire, application of the FASS, and physical examination using the Pelvic Prgan Prolapse Quantification (POP-Q) system. The translation and cross-cultural adaptation were performed following the international methodology. The psychometric properties tested were criterion validity, construct validity, stability, and reliability. For this purpose, the comparison with POP-Q findings and between symptomatic and asymptomatic patients, test-retest and internal consistency (Cronbach's alpha) were used, respectively. The intraclass correlation coefficient (ICC) was calculated to assess the level of agreement between evaluations (inter- and intraobservers). P < 0.05 was considered statistically significant. RESULTS: The calculated Cronbach's alpha coefficient was 0.76, indicating strong reliability for the validation sample. Symptomatic women had different scores on all FASS items as well as total score when compared with asymptomatic women (p < 0.001). Intraobserver coefficient ranged from 0.91 (urinary symptoms) to 0.98 (FASS total score), indicating excellent concordance level in all items. Interobserver coefficient ranged from 0.47 (intestinal symptoms) to 0.90 (FASS total score), indicating moderate to excellent correlation. CONCLUSIONS: The psychometric properties tested in the FASS Portuguese version proved to be a valid and reliable for evaluating signs and symptoms related to pelvic floor function in Brazilian women.

Microbial-derived products as potential new antimicrobials.

Due to the continuing global concerns involving antibiotic resistance, there is a need for scientific forums to assess advancements in the development of antimicrobials and their alternatives that might reduce development and spread of antibiotic resistance among bacterial pathogens. The objectives of the 2nd International Symposium on Alternatives to Antibiotics were to highlight promising research results and novel technologies that can provide alternatives to antibiotics for use in animal health and production, assess challenges associated with their authorization and commercialization for use, and provide actionable strategies to support their development. The session on microbial-derived products was directed at presenting novel technologies that included exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials, probiotics development via fecal microbiome transplants among monogastric production animals such as chickens and mining microbial sources such as bacteria or yeast to identify new antimicrobial compounds. Other research has included continuing development of antimicrobial peptides such as newly discovered bacteriocins as alternatives to antibiotics, use of bacteriophages accompanied by development of unique lytic proteins with specific cell-wall binding domains and novel approaches such as microbial-ecology guided discovery of anti-biofilm compounds discovered in marine environments. The symposium was held at the Headquarters of the World Organisation for Animal Health (OIE) in Paris, France during 12-15 December 2016.

How to use the Pelvic Organ Prolapse Quantification (POP-Q) system?

AIMS: To set out the basic description of pelvic organ prolapse (POP) using the International Continence Society/International Urogynecology Association Pelvic Organ Prolapse Quantification (POP-Q) system. METHODS: The basic approach to use of the POP-Q was identified and summarized. RESULTS: Six defined points in the vagina are identified; points Aa and Ba for the anterior vagina, Ap and Bp for the posterior vagina, and C and D for the cervix/vault. Point D is not used in women who previously had a hysterectomy. The patient is asked to strain, ideally when in the standing position, to elicit the POP to its maximum extent. The location of the defined points is then gauged relative to the hymenal ring and recorded on a grid. Three additional measurements are taken to achieve a full description; the genital hiatus length, perineal body length, and total vaginal length. Staging a POP relies on identifying the lowest extent of any part of the six defined points; if any point reaches close to the hymenal ring (at least stage 2), the prolapse is usually symptomatic. CONCLUSIONS: The POP-Q system is readily cataloged and offers detailed description of considerable benefit in clinical practice and research.

Informed consent checklists for midurethral slings: a common-sense approach.

INTRODUCTION AND HYPOTHESIS: Following the US Food and Drug Administration's (FDA's) warning about the use of transvaginal mesh to treat pelvic organ prolapse (POP) and the use of single-incision slings to treat incontinence, the number of lawsuits for medical negligence regarding the use of any polypropylene mesh in the vagina has increased tremendously. METHODS: This same FDA document did not question the use of polypropylene midurethral slings and polypropylene for sacrocolpopexies. Surprisingly, despite all the evidence and recommendations from respected international scientific societies, we are constantly being called upon by our patients to defend the use of midurethral slings. The most common reasons for the new rash of medicolegal proceedings involving midurethral slings has to do with "breach of duties" resulting from undisclosed postoperative complications on the consent form and/or the lack of information in the medical records confirming that all possible alternative treatment options were presented to and discussed with the patient. RESULTS: One response to these lawsuits involves the addition of preoperative checklists when performing informed consent with patients electing surgical correction of stress urinary incontinence (SUI). CONCLUSIONS: This clinical opinion provides an expert clinician's perspectives and legal point of view on this controversial topic and discusses the role of a preoperative checklist supplementary to the standard informed consent form.