Evidence for increased tone or overactivity of pelvic floor muscles in pelvic health conditions: a systematic review.

OBJECTIVE: Pelvic floor muscle tone, which includes active and passive components, is argued to be increased in many pelvic health conditions, including those involving pain. This study systematically reviewed evidence for increased pelvic floor muscle tone in pelvic health conditions. DATA SOURCES: Electronic databases (PubMed, CINAHL, and Embase) were searched up to May 31, 2021. The search strategy included variants of pelvic and/or floor, muscle, and tone using keywords and Medical Subject Headings (MeSH) terms. STUDY ELIGIBILITY CRITERIA: Studies were included if they investigated increased tone of the pelvic floor muscle and reported measures of active or mechanical properties of the pelvic floor muscle in humans with any pelvic health condition, including pain, bowel, urogenital, or sexual dysfunctions. Studies of any design were included, except systematic and narrative reviews. Reference lists of studies, reviews, and book chapters were searched for additional studies. METHODS: Data were extracted using a standardized form, including measurement tool and outcome measure. Risk of bias was analyzed using a modified ROBINS-I (Risk of Bias In Non-randomized Studies - of Interventions) tool, and a score was allocated to determine whether the study provided "convincing" interpretation (comparison with condition-free control group, valid measure, no application issues). RESULTS: In total, 151 studies were included, reporting 8 different tools (electromyography, dynamometry, manometry, digital palpation, defecography, ultrasound, magnetic resonance imaging, other). The most common pelvic health condition was pelvic pain (n=16 conditions), followed by bowel and urogenital conditions. Most studies (57%) were cross-sectional. A healthy control group was infrequently included for comparison (27%). Unvalidated methods or methods applied in a manner that precluded convincing interpretation were common (94%). Of the 15 measurement tools that provided convincing evidence, 10 demonstrated greater tone in a pelvic health condition (all pain) compared with controls, and 5 showed no difference. CONCLUSION: Despite the large literature, few studies provide convincing evidence for increased tone/overactivity of pelvic floor muscles in pelvic health conditions. Interpretation is hampered by design and measurement issues. Terminology was often inaccurate. Few studies investigate male, transgender, and pediatric groups.

The influence of prostatectomy and body position on location and displacement of pelvic landmarks with pelvic floor muscle contraction.

AIMS: To compare pelvic floor muscle (PFM) anatomy and function (i) between pre- and post-prostatectomy in standing, and (ii) between sitting and standing postprostatectomy. METHODS: Thirty-two men scheduled to undergo a prostatectomy volunteered to participate. Transperineal ultrasound imaging was used to visualize five anatomical pelvic landmarks that have been validated to reflex anatomy and activity of PFMs (pubic symphysis, anorectal junction [ARJ], mid-urethra [MU], bulb of penis [BP], and urethrovesical junction [UVJ]). Both before and after prostatectomy, participants performed three submaximal PFM contractions in sitting and/or standing positions while ultrasound data were recorded. RESULTS: Postprostatectomy the UVJ location was more caudal and dorsal, the ARJ (puborectalis) vector was longer, the BP was more ventral than preprostatectomy, and these landmarks moved less ventrally with contraction. After prostatectomy, the MU, BP, and ARJ were more ventral in standing than sitting. The UVJ was more caudal and elevated more with contraction in standing than sitting after prostatectomy. CONCLUSION: These data demonstrate differences in the anatomy and mechanics of PFMs post- versus pre-prostatectomy, and between sitting and standing positions postprostatectomy. Findings are consistent with surgical changes to the bladder and urethral anatomy. Reduced passive support for the urethra and bladder are likely to may contribute to differences between standing and sitting postprostatectomy.

The relationship between pre- and postprostatectomy measures of pelvic floor muscle function and development of early incontinence after surgery.

AIMS: The aim of this study is to investigate (i) whether pelvic floor muscle (PFM) shortening can be enhanced by provision of training focused on striated urethral sphincter (SUS) with feedback before prostatectomy, (ii) whether PFM shortening during voluntary efforts and coughing before and after prostatectomy differs between men who do and do not report symptoms of urinary incontinence 1 month after prostatectomy, and (iii) the relationship between severity of incontinence after prostatectomy and features of pelvic floor function (muscle shortening) and urethral length before and after prostatectomy. METHODS: Sixty men referred for preoperative PFM training before radical prostatectomy participated. The International Continence Society Male Short Form questionnaire was used to quantify continence status. Transperineal ultrasound (US) imaging was used to record pelvic displacements related to activation of striated urethral sphincter, bulbocavernosus (BC) and puborectalis muscles during cough, "natural" voluntary contraction following pamphlet instruction, and trained voluntary contraction after formal physiotherapist instruction including US feedback. RESULTS: Pelvic floor displacements following training differed between continent and incontinent men; continent participants demonstrated increased SUS shortening after training (compared with "natural"), but no difference was observed between trained and "natural" contractions for incontinent participants. Motion at ano-rectal junction during cough was reduced following surgery, but voluntary and involuntary activation of SUS or BC was not consistently affected by surgery. CONCLUSIONS: Participants' capacity to improve function of the SUS with training appears related to postprostatectomy continence outcome.

Validation of shear wave elastography as a noninvasive measure of pelvic floor muscle stiffness.

OBJECTIVES: To investigate the validity of shear wave elastography (SWE) as a measure of stiffness of the puborectalis muscle by examining: (1) the relationship between puborectalis muscle stiffness and pelvic floor muscle (PFM) activation at different intensities; and (2) the relationship between puborectalis stiffness and pelvic floor morphometry during contractions at different intensities. METHODS: Fifteen healthy asymptomatic women performed 6-s isometric PFM contractions at different intensities (0, 10%, 20%, 30%, 50%, 75%, and 100% of maximal voluntary contraction) guided by intravaginal electromyography (EMG). Stiffness of the puborectalis muscle was measured using SWE by calculating the average shear modulus in regions of interest that contained puborectalis muscle fibers parallel to the transducer. Pelvic floor morphometry was assessed in the mid-sagittal plane using transperineal B-mode ultrasound imaging. Shear modulus, EMG (root mean square amplitude) and pelvic floor morphometry parameters were normalized to the value recorded during maximal voluntary contraction. To assess the relationship between stiffness and pelvic floor activation/morphometry, coefficient of determination (r2 ) was calculated for each participant and a group average was computed. RESULTS: Shear modulus and EMG were highly correlated (average r2 ; left 0.90 ± 0.08, right 0.87 ± 0.15). Shear modulus also strongly correlated with bladder neck position (x-axis horizontal coordinates relative to the pubic symphysis), anorectal rectal angle and position, levator plate angle, and antero-posterior diameter of the levator hiatus (average r2 : range 0.62-0.78). CONCLUSIONS: These findings support the validity of SWE to assess puborectalis muscle stiffness in females. Stiffness measures were strongly associated with PFM EMG and pelvic floor morphometry and may be used to indirectly assess the level of activation of the puborectalis muscle without the use of more invasive techniques. By overcoming limitations of current assessment tools, this promising noninvasive and real-time technique could enable important breakthrough in the pathophysiology and management of pelvic floor disorders.

The repeatability of measurements of male pelvic floor anatomy and function made from transperineal ultrasound images of healthy men and those before and after prostatectomy.

AIMS: To investigate the inter- and intratester repeatability of measurement of the location and displacement of five pelvic landmarks related to pelvic floor muscles with transperineal ultrasound (TPUS) imaging recorded from healthy men and men before and after prostatectomy. METHODS: TPUS images were selected from four different participant groups: healthy men, men awaiting prostatectomy, men 2 weeks after prostatectomy, and men 12 months after prostatectomy. On two separate occasions, two assessors with different levels of experience performed analysis of location and displacement of five pelvic landmarks in images made at rest and during voluntary contraction. A two-way mixed effects, single measurement, absolute agreement intraclass correlation coefficient (ICC) was used to investigate the repeatability. RESULTS: Intertester reliability of all locations at rest for all groups was excellent (ICCs > 0.8) except for the craniocaudal coordinate of the ventral urethrovesical junction for men 2 weeks postprostatectomy and the anorectal junction for men with a cancerous prostate. Intertester reliability of the measurement of landmark displacement was acceptable (>0.5) for the dorsoventral axis of motion but not for the craniocaudal axis of motion for all landmarks across all groups. The more experienced assessor was consistently more repeatable. More deeply placed landmarks were more often excluded from analysis and had poorer reliability. CONCLUSIONS: Analysis of TPUS images across clinical groups is repeatable for both location and displacement of pelvic landmarks related to pelvic floor muscles when measures are made twice. Analysis experience, landmark depth and optimization of ultrasound settings appear to be important factors in reliability.

Influence of body position on dynamics of the pelvic floor measured with transperineal ultrasound imaging in men.

AIMS: This paper aims to evaluate the feasibility of transperineal ultrasound imaging (TPUS) for visualizing the motion of pelvic landmarks associated with striated pelvic floor muscle contraction in men in standing; to compare the locations of pelvic landmarks between sitting and standing; and to compare the effects of different body positions on measures of pelvic floor muscle contraction. METHODS: Thirty-five men awaiting prostatectomy volunteered to participate. Participants performed three repetitions of submaximal pelvic floor contraction in sitting and again in standing. Movement of pelvic landmarks with contraction was recorded using an ultrasound imaging transducer placed on the perineum. RESULTS: The feasibility of TPUS in men in standing was demonstrated through the visualization of three out of four pelvic landmarks in more than 95% of images in the standing position. Analysis of pelvic landmarks and their respective relationships with muscle shortening demonstrated that the anorectal junction and urethrovesical junction were lower and the estimated length of puborectalis was shorter in standing than sitting. The mid-urethra (striated urethral sphincter) and anorectal junction (puborectalis) landmark displaced further cranially in standing than sitting. CONCLUSIONS: TPUS can be used to visualize three pelvic landmarks in men with cancerous prostates. Puborectalis is shorter at rest in standing than sitting, and elevation of the mid-urethra and the anorectal junction is more in standing than sitting. Together these findings indicate that feedback for pelvic floor muscle training is possible in both positions, but the position needs to be standardized for a comparative assessment.

Comparison of dynamic features of pelvic floor muscle contraction between men with and without incontinence after prostatectomy and men with no history of prostate cancer.

AIM: To compare features of pelvic floor muscle function between men with and without incontinence after prostatectomy and men with no history of prostate cancer. METHODS: The study included men with incontinence postprostatectomy (PPI; n = 20), continent men postprostatectomy (PPC; n = 23) and a control group (CC; n = 20). Transperineal ultrasound imaging recorded motion associated with contraction of the striated urethral sphincter (SUS), puborectalis (PR) and bulbocavernosus (BC) muscles during maximal voluntary contraction (MVC), submaximal efforts, evoked coughing and bearing down. Anatomical landmark displacements were compared between groups and receiver operating characteristics were calculated to determine the threshold displacements that best differentiated PPI and PPC. RESULTS: PPC demonstrated greater SUS, PR, and BC displacement than PPI during MVC (All: P < .01). During cough, PPC had less bladder neck descent (PR lengthening), and greater BC shortening (P = .003) than both PPI and CC. PPC also achieved greater SUS displacement (P = .025) than PPI during cough. The best discrimination between PPI and PPC was achieved when men exceeded threshold displacement for both SUS (≥4.1 mm) and PR (≥2.4 mm) during MVC. The urethral length was not different between PPC and PPI. CONCLUSIONS: Men who were continent postprostatectomy achieved greater shortening of the SUS, PR, and BC muscles than incontinent men during voluntary contractions and demonstrated better PR and BC function than control participants during coughing. The capacity to shorten the SUS ≥4.1 mm and the PR ≥2.4 mm best distinguished between PPI and PPC and might be a useful clinical target for conservative treatment programs.

Activity of Deep and Superficial Pelvic Floor Muscles in Women in Response to Different Verbal Instructions: A Preliminary Investigation Using a Novel Electromyography Electrode.

INTRODUCTION: Verbal instructions are used clinically to encourage activation of the pelvic floor muscles (PFM). Whether separate layers of PFM activate differently in response to instructions remains unknown. AIM: To test the hypotheses that (i) instructions that aimed to bias activity of a specific muscle layer would increase activation of the targeted layer to a greater extent than the other layer, (ii) activity of individual PFM layers would differ between instructions, and (iii) PFM activity would be symmetrical for all instructions. METHOD: PFM electromyography (EMG) was recorded using custom-designed surface electrodes in 12 women without PFM dysfunction. The electrode included 4 pairs of recording surfaces orientated to measure EMG from deep and superficial PFM on each side. 3 submaximal contractions were performed for 5 seconds in response to 7 verbal instructions. Root-mean-squared EMG amplitude was calculated for 1 second during the period when participants most closely matched the target activation level. A repeated-measures anova was used to test whether PFM EMG differed between instructions and between regions. The EMG increase of individual muscles relative to that of the reference muscle [deep/right PFM] was compared to no change with t-tests for single samples. MAIN OUTCOME MEASURE: PFM EMG amplitude. RESULTS: Superficial PFM EMG was greater than deep PFM for all instructions (P = .039). 2 instructions induced the greatest amplitude of EMG for the superficial PFM: "squeeze the muscles around the vaginal opening as if to purse lips of your mouth" and "draw the clitoris in a posterior direction" (P = .036). Asymmetry was found in the deeper PFM in 3 instructions designed to bias the superficial PFM. STRENGTH & LIMITATIONS: This preliminary study recorded activation of deep and superficial PFM layers in females with a custom-designed novel electrode. Some cross-talk of recording between muscle layers is possible but unlikely to impact the major findings. CONCLUSION: Verbal instructions used to teach PFM contractions can influence their pattern of activity. This study provides preliminary evidence that, in a selection of verbal instructions, the superficial PFM activates more than the deep PFM, and that the deep PFM can have asymmetrical activation. Aljuraifani R, Stafford RE, Hall LM, et al. Activity of Deep and Superficial Pelvic Floor Muscles in Women in Response to Different Verbal Instructions: A Preliminary Investigation Using a Novel Electromyography Electrode J Sex Med 2019;16:673-679.

Female striated urogenital sphincter contraction measured by shear wave elastography during pelvic floor muscle activation: Proof of concept and validation.

AIMS: Investigation of the function of the striated urogenital sphincter (SUS) is challenging because it is difficult to access and requires invasive measures. Ultrasound shear wave elastography (SWE) is a non-invasive real-time technique used to estimate tissue stiffness. As muscle stiffness can be used as an estimate of muscle force, SWE provides an opportunity to study contraction of the peri-urethral musculature. Validation of SWE to study SUS during functional tasks, such as pelvic floor muscle contractions, is required prior to application in clinical populations. METHODS: Ten healthy females (34[5] years) participated. Stiffness in a region expected to contain the SUS was quantified using SWE at rest and during a pelvic floor muscle contractions performed at 10%, 25%, and 50% of maximal voluntary contraction (MVC). Two repetitions were performed for 10 s. RESULTS: During contraction, stiffness increased in the region of the SUS in all participants and at all contraction intensities. Multiple regions of increased stiffness were detected, with 95.8% of regions situated ventral to the mid-urethra within the anatomical area of the SUS. The increase in stiffness was greater for 50% MVC than both 10% and 25% MVC contraction intensities (P < 0.01). CONCLUSIONS: Stiffness increased within the anatomical region of the SUS during voluntary pelvic floor muscle contractions with predictable response to changes in contraction intensity. These observations support the potential for ultrasound SWE to study SUS function non-invasively.

Postprostatectomy incontinence is related to pelvic floor displacements observed with trans-perineal ultrasound imaging.

AIMS: To investigate the relationship between post-prostatectomy incontinence and dynamic features of activation of specific pelvic floor muscles in addition to anatomical parameters of the urethra. METHODS: Forty-two men aged 66 (7) years (incontinent [N = 19] and continent [N = 23]) who had undergone prostatectomy participated. Transperineal ultrasound imaging was used to record sagittal images of pelvic structures during involuntary coughing and sustained maximal voluntary contractions. Imaging data were analyzed to calculate displacements of pelvic floor landmarks associated with activation of the puborectalis, striated urethral sphincter, and bulbocavernosus muscles. Anatomical features of functional urethral length and the resting position of the ano-rectal and urethra-vesical junctions were calculated. A principal component analysis and multiple logistic regression were used to consider which combinations of variables best distinguish between men with and without incontinence. RESULTS: Five principal components were identified that together explained 72.0% of the data. Two principal components that represented (i) striated urethral sphincter activation and (ii) bulbocavernosus and puborectalis muscle activation were significantly different between participants with and without incontinence. Together these components correctly identified 88.1% of incontinent men, with a specificity and sensitivity of 91.3% and 84.2%, respectively. Poor function of the bulbocavernosus and puborectalis muscles could be compensated by good striated urethral sphincter function, but the bulbocavernosus and puborectalis muscles had less potential to compensate for poor striated urethral sphincter function. CONCLUSIONS: Dynamic features of pelvic floor muscle activation, particularly shortening of the striated urethral sphincter during cough and voluntary contraction, are related to continence status after prostatectomy.

Application of shear-wave elastography to estimate the stiffness of the male striated urethral sphincter during voluntary contractions.

OBJECTIVES: To investigate whether increases in stiffness can be detected in the anatomical region associated with the striated urethral sphincter (SUS) during voluntary activation using shear-wave elastography (SWE); to identify the location and area of the stiffness increase relative to the point of greatest dorsal displacement of the mid urethra (i.e. SUS); and to determine the relationship between muscle stiffness and contraction intensity. SUBJECTS AND METHODS: In all, 10 healthy men participated. A linear ultrasound (US) transducer was placed mid-sagittal on the perineum adjacent to a pair of electromyography electrodes that recorded non-specific pelvic floor muscle activity. Stiffness in the area expected to contain the SUS was estimated via US SWE at rest and during voluntary pelvic floor muscles contractions to 5%, 10% and 15% maximum. Still image frames were exported for each repetition and analysed with software that detected increases in stiffness above 150% of the resting stiffness. RESULTS: Pelvic floor muscle contraction elicited an increase in stiffness above threshold within the region expected to contain the SUS for all participants and contraction intensities. The mean (SD) ventral-dorsal distance between the centre of the stiffness area and region of maximal motion of the mid-urethra (caused by SUS contraction) was 5.6 (1.8), 6.2 (0.8), and 5.8 (0.7) mm for 5%, 10% and 15% maximal voluntary contraction, respectively. Greater pelvic floor muscle contraction intensity resulted in a concomitant increase in stiffness, which differed between contraction intensities (5% vs 10%, P < 0.001; 5% vs 15%, P < 0.001; 10% vs 15%, P = 0.003). CONCLUSION: Voluntary contraction of the pelvic floor muscles in men is associated with an area of stiffness increase measured with SWE, which concurs with the expected location of the SUS. The increase in stiffness occurred in association with an increase in perineal surface electromyography activity, providing evidence that stiffness amplitude relates to general pelvic floor muscle contraction intensity. Future applications of SWE may include investigations of patient populations in which dysfunction of the SUS is thought to play an important role, or investigation of the effect of rehabilitation programmes that target this muscle.

Pattern of activation of pelvic floor muscles in men differs with verbal instructions.

AIMS: To investigate the effect of instruction on activation of pelvic floor muscles (PFM) in men as quantified by transperineal ultrasound imaging (US) and to validate these measures with invasive EMG recordings. METHODS: Displacement of pelvic floor landmarks on transperineal US, intra-abdominal pressure (IAP) recorded with a nasogastric transducer, and surface EMG of the abdominal muscles and anal sphincter were recorded in 15 healthy men during sub-maximal PFM contractions in response to different verbal instructions: "tighten around the anus," "elevate the bladder," "shorten the penis," and "stop the flow of urine." In three men, fine-wire EMG recordings were made from puborectalis and bulbocavernosus, and trans-urethral EMG recordings from the striated urethral sphincter (SUS). Displacement data were validated by analysis of relationship with invasive EMG. Displacement, IAP, and abdominal/anal EMG were compared between instructions. RESULTS: Displacement of pelvic landmarks correlated with the EMG of the muscles predicted anatomically to affect their locations. Greatest dorsal displacement of the mid-urethra and SUS activity was achieved with the instruction "shorten the penis." Instruction to "elevate the bladder" induced the greatest increase in abdominal EMG and IAP. "Tighten around the anus" induced greatest anal sphincter activity. CONCLUSIONS: The pattern of urethral movement measured from transperineal US is influenced by the instructions used to teach activation of the pelvic floor muscles in men. Efficacy of PFM training may depend on the instructions used to train activation. Instructions that optimize activation of muscles with a potential to increase urethral pressure without increasing abdominal EMG/IAP are likely ideal. Neurourol. Urodynam. 35:457-463, 2016. © 2015 Wiley Periodicals, Inc.

A general sequence processing and analysis program for protein engineering.

Protein engineering projects often amass numerous raw DNA sequences, but no readily available software combines sequence processing and activity correlation required for efficient lead identification. XLibraryDisplay is an open source program integrated into Microsoft Excel for Windows that automates batch sequence processing via a simple step-by-step, menu-driven graphical user interface. XLibraryDisplay accepts any DNA template which is used as a basis for trimming, filtering, translating, and aligning hundreds to thousands of sequences (raw, FASTA, or Phred PHD file formats). Key steps for library characterization through lead discovery are available including library composition analysis, filtering by experimental data, graphing and correlating to experimental data, alignment to structural data extracted from PDB files, and generation of PyMOL visualization scripts. Though larger data sets can be handled, the program is best suited for analyzing approximately 10 000 or fewer leads or naïve clones which have been characterized using Sanger sequencing and other experimental approaches. XLibraryDisplay can be downloaded for free from sourceforge.net/projects/xlibrarydisplay/ .

Vocoder simulations of highly focused cochlear stimulation with limited dynamic range and discriminable steps.

OBJECTIVES: There is recent interest in focused stimulation of the cochlea via modalities such as tripolar electrical and infrared neural stimulation to improve speech in noise comprehension and music perception. The purpose of this work was to use vocoder-based simulations to investigate speech recognition for broad stimulation (standard monopolar paradigm) versus more focused stimulation under a variety of signal-to-noise ratios, dynamic ranges, and numbers of discriminable loudness steps. DESIGN: Vocoder simulations were used to assess the intelligibility of sentences, consonants, and vowels that were noise vocoded and presented to 7 normal-hearing listeners for identification. A novel aspect of the simulations presented here was the use of nonuniform quantization steps within the dynamic range to more closely simulate the Weber functions observed in cochlear implant users. Intelligibility was assessed for the different filter slopes under a variety of signal-to-noise ratio levels, dynamic ranges, and numbers of discriminable steps. RESULTS: Speech processed via vocoder simulations representing focused stimulation was found to be substantially more intelligible than speech processed via a monopolar electric vocoder simulation, with differences of up to 60 percentage points. There were no significant differences, however, seen between the two focused approaches (signal attenuations of 10 and 17 dB/mm) for the conditions investigated. Speech processed via the highly focused vocoder (17 dB/mm) was robust to constraints on small envelope dynamic range and small number of discriminable steps within the dynamic range, as high performance was maintained with at least a 5 dB dynamic range and eight or more discriminable steps. Significant drops in intelligibility were noted when the number of steps fell below eight. CONCLUSIONS: Highly focused stimulation-tripolar electrical and infrared neural stimulation-has potential for increased performance in noise compared with monopolar stimulation, but much work remains to bear this potential out and to take full advantage of each modality's strengths.

Potential role of physical labor and cultural views of menstruation in high incidence of pelvic organ prolapse in Nepalese women: a comparative study across the menstrual cycle.

Introduction: Pelvic organ prolapse (POP) is a significant health concern for young Nepali women, with potential risk factors including pelvic floor trauma from vaginal delivery and heavy lifting. The prevalence of symptomatic POP (SPOP) among nulliparous women in Nepal is 6%, while the general population of Nepali women aged 15-49 years reports a prevalence of 7%. Surprisingly, the average age of SPOP onset in Nepal is 27 years, challenging the assumption that postmenopausal age and vaginal delivery are the sole risk factors. This study aims to investigate the influence of increased intra-abdominal pressure (IAP) during lifting tasks on pelvic organ descent in Nepali women across different menstrual cycle stages. Methods: The study included 22 asymptomatic Nepali women aged 18-30 years who regularly engage in heavy lifting. Intra-abdominal pressure was measured intra-vaginally during typical and simulated lifting tasks, which encompassed various scenarios such as ballistic lifting, ramped lifting, and pre-contraction of pelvic floor muscles, as well as coughing, Valsalva maneuver, and pelvic floor contractions. Pelvic floor displacement was recorded using transperineal ultrasound during menstruation, ovulation, and the mid-luteal phase. Results: Results indicated that pelvic floor displacement was greater during menstruation than ovulation when performing a simulated ballistic lifting task (6.0 ± 1.6 mm vs. 5.1 ± 1.5 mm, p = 0.03, d = 0.6). However, there was no significant difference in pelvic floor displacement during lifting when the pelvic muscles were pre-contracted. Conclusion: These findings suggest that lifting heavy loads during menstruation may increase the risk of stretching and injuring pelvic floor supportive tissues, potentially contributing to SPOP in young Nepali women. Pre-contracting pelvic floor muscles during lifting tasks may offer a protective effect. Understanding these factors could aid in developing targeted preventive measures and raising awareness about the impact of heavy lifting on pelvic floor health among Nepali women.

Pelvic floor muscle length changes with breathing in males: A preliminary report.

The study aimed to identify whether pelvic floor muscles modulate length with breathing, and if any length changes induced by breathing relate to abdominal cavity displacement and intra-abdominal pressure. To investigate these relationships, displacement of pelvic landmarks that related to pelvic floor muscle length using transperineal ultrasound imaging, breath volume, intra-abdominal pressure, abdominal and ribcage displacement, and abdominal and anal sphincter muscle electromyography were measured during quiet breathing and breathing with increased dead-space in ten healthy men. Pelvic floor muscle landmark displacement modulated with ribcage motion during breathing. This relationship was stronger for: i) motion of the urethrovesical junction (puborectalis muscle length change) than the mid-urethra landmark (striated urethral sphincter muscle length change), and ii) dead-space breathing in standing than dead-space breathing in supine or quiet breathing in standing. In most (but not all) participants, the urethrovesical junction descended during inspiration and elevated during expiration. Striated urethral sphincter length changes during the respiratory cycle was independent of intra-abdominal pressure. In summary, breathing involves pelvic floor muscle length changes and is consistent with the role of these muscles during respiration to aid maintenance of continence, lung ventilation and/or provision of support to the abdominal cavity. Clinicians who train pelvic floor muscles need to be aware that length change of pelvic floor muscles is expected with breathing.

A comparison of the binding sites of antibodies and single-domain antibodies.

Antibodies are the largest class of biotherapeutics. However, in recent years, single-domain antibodies have gained traction due to their smaller size and comparable binding affinity. Antibodies (Abs) and single-domain antibodies (sdAbs) differ in the structures of their binding sites: most significantly, single-domain antibodies lack a light chain and so have just three CDR loops. Given this inherent structural difference, it is important to understand whether Abs and sdAbs are distinguishable in how they engage a binding partner and thus, whether they are suited to different types of epitopes. In this study, we use non-redundant sequence and structural datasets to compare the paratopes, epitopes and antigen interactions of Abs and sdAbs. We demonstrate that even though sdAbs have smaller paratopes, they target epitopes of equal size to those targeted by Abs. To achieve this, the paratopes of sdAbs contribute more interactions per residue than the paratopes of Abs. Additionally, we find that conserved framework residues are of increased importance in the paratopes of sdAbs, suggesting that they include non-specific interactions to achieve comparable affinity. Furthermore, the epitopes of sdAbs are only marginally less accessible than those of Abs: we posit that this may be explained by differences in the orientation and compaction of sdAb and Ab CDR-H3 loops. Overall, our results have important implications for the engineering and humanization of sdAbs, as well as the selection of the best modality for targeting a particular epitope.

Targeting CD74 in B-cell non-Hodgkin lymphoma with the antibody-drug conjugate STRO-001.

Overexpression of CD74, a type II transmembrane glycoprotein involved in MHC class II antigen presentation, has been reported in many B-cell non-Hodgkin lymphomas (NHLs) and in multiple myeloma (MM). STRO-001 is a site-specific, predominantly single-species antibody-drug conjugate (ADC) that targets CD74 and has demonstrated efficacy in xenograft models of MM and tolerability in non-human primates. Here we report results of preclinical studies designed to elucidate the potential role of STRO-001 in B-cell NHL. STRO-001 displayed nanomolar and sub-nanomolar cytotoxicity in 88% (15/17) of cancer cell lines tested. STRO-001 showed potent cytotoxicity on proliferating B cells while limited cytotoxicity was observed on naïve human B cells. A linear dose-response relationship was demonstrated in vivo for DLBCL models SU-DHL-6 and U2932. Tumor regression was induced at doses less than 5 mg/kg, while maximal activity with complete cures were observed starting at 10 mg/kg. In MCL Mino and Jeko-1 xenografts, STRO-001 starting at 3 mg/kg significantly prolonged survival or induced tumor regression, respectively, leading to tumor eradication in both models. In summary, high CD74 expression levels in tumors, nanomolar cellular potency, and significant anti-tumor in DLBCL and MCL xenograft models support the ongoing clinical study of STRO-001 in patients with B-cell NHL.

Optimization of a Glucagon-Like Peptide 1 Receptor Antagonist Antibody for Treatment of Hyperinsulinism.

Congenital hyperinsulinism (HI) is a genetic disorder in which pancreatic ß-cell insulin secretion is excessive and results in hypoglycemia that, without treatment, can cause brain damage or death. Most patients with loss-of-function mutations in ABCC8 and KCNJ11, the genes encoding the ß-cell ATP-sensitive potassium channel (KATP), are unresponsive to diazoxide, the only U.S. Food and Drug Administration-approved medical therapy and require pancreatectomy. The glucagon-like peptide 1 receptor (GLP-1R) antagonist exendin-(9-39) is an effective therapeutic agent that inhibits insulin secretion in both HI and acquired hyperinsulinism. Previously, we identified a highly potent antagonist antibody, TB-001-003, which was derived from our synthetic antibody libraries that were designed to target G protein-coupled receptors. Here, we designed a combinatorial variant antibody library to optimize the activity of TB-001-003 against GLP-1R and performed phage display on cells overexpressing GLP-1R. One antagonist, TB-222-023, is more potent than exendin-(9-39), also known as avexitide. TB-222-023 effectively decreased insulin secretion in primary isolated pancreatic islets from a mouse model of hyperinsulinism, Sur1-/- mice, and in islets from an infant with HI, and increased plasma glucose levels and decreased the insulin to glucose ratio in Sur1-/- mice. These findings demonstrate that targeting GLP-1R with an antibody antagonist is an effective and innovative strategy for treatment of hyperinsulinism. ARTICLE HIGHLIGHTS: Patients with the most common and severe form of diazoxide-unresponsive congenital hyperinsulinism (HI) require a pancreatectomy. Other second-line therapies are limited in their use because of severe side effects and short half-lives. Therefore, there is a critical need for better therapies. Studies with the glucagon-like peptide 1 receptor (GLP-1R) antagonist, avexitide (exendin-(9-39)), have demonstrated that GLP-1R antagonism is effective at lowering insulin secretion and increasing plasma glucose levels. We have optimized a GLP-1R antagonist antibody with more potent blocking of GLP-1R than avexitide. This antibody therapy is a potential novel and effective treatment for HI.

Discovery of STRO-002, a Novel Homogeneous ADC Targeting Folate Receptor Alpha, for the Treatment of Ovarian and Endometrial Cancers.

STRO-002 is a novel homogeneous folate receptor alpha (FolRα) targeting antibody-drug conjugate (ADC) currently being investigated in the clinic as a treatment for ovarian and endometrial cancers. Here, we describe the discovery, optimization, and antitumor properties of STRO-002. STRO-002 was generated by conjugation of a novel cleavable 3-aminophenyl hemiasterlin linker-warhead (SC239) to the nonnatural amino acid para-azidomethyl-L-phenylalanine incorporated at specific positions within a high affinity anti-FolRα antibody using Sutro's XpressCF+, which resulted in a homogeneous ADC with a drug-antibody ratio (DAR) of 4. STRO-002 binds to FolRα with high affinity, internalizes rapidly into target positive cells, and releases the tubulin-targeting cytotoxin 3-aminophenyl hemiasterlin (SC209). SC209 has reduced potential for drug efflux via P-glycoprotein 1 drug pump compared with other tubulin-targeting payloads. While STRO-002 lacks nonspecific cytotoxicity toward FolRα-negative cell lines, bystander killing of target negative cells was observed when cocultured with target positive cells. STRO-002 is stable in circulation with no change in DAR for up to 21 days and has a half-life of 6.4 days in mice. A single dose of STRO-002 induced significant tumor growth inhibition in FolRα-expressing xenograft models and patient-derived xenograft models. In addition, combination treatment with carboplatin or Avastin further increased STRO-002 efficacy in xenograft models. The potent and specific preclinical efficacy of STRO-002 supports clinical development of STRO-002 for treating patients with FolRα-expressing cancers, including ovarian, endometrial, and non-small cell lung cancer. Phase I dose escalation for STRO-002 is in progress in ovarian cancer and endometrial cancer patients (NCT03748186 and NCT05200364).