Translabial Ultrasound: An Effective Modality for Evaluation of Midurethral Sling Revision.

ABSTRACT: Midurethral sling (MUS) surgical procedures, in which a polypropylene synthetic mesh graft is transvaginally placed to support the urethra and manage stress urinary incontinence (SUI), is commonly performed ( Urology. 2013;82(1):38-41; Radiology. 2018;289(3):728-729). Within 10 years of sling placement, about 1 in 20 women undergo subsequent surgery to revise their mesh, in which the mesh is cut or partially excised ( Obstet Gynecol . 2019;133:1099-1108). Translabial ultrasound (TLUS) has been described as a sensitive technique for viewing MUS, although few studies have evaluated its ability to view surgically revised MUS ( Radiology . 2018;289(3):721-727). Understanding the anatomy of the MUS is critical to urologists and urogynecologists striving to optimize management of patients presenting with lower urinary tract symptoms and history of MUS. To assess the clinical utility and reliability of TLUS as a diagnostic tool in its detection of MUS discontinuity, we conducted a retrospective analysis on patients who underwent TLUS at a tertiary care center between September 2017 and May 2020 for indication of lower urinary tract symptoms and history of MUS placement. Performance of TLUS was evaluated by comparing findings with operative or clinical records. Among the 81 women included, detection of MUS revision, which was defined as a discontinuity in sling material, had a sensitivity of 84.6% and specificity of 97.1%. Translabial ultrasound is an inexpensive, nonirradiating, and noninvasive modality that is effective at visualizing MUSs. It is a reliable identifier of previous MUS revision, in which it detects a midline discontinuity of the hyperechoic mesh with an average 10 mm gap.

A small-molecule screen reveals novel modulators of MeCP2 and X-chromosome inactivation maintenance.

BACKGROUND: Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in the X-linked methyl-CpG binding protein 2 (MeCP2) gene. While MeCP2 mutations are lethal in most males, females survive birth but show severe neurological defects. Because X-chromosome inactivation (XCI) is a random process, approximately 50% of the cells silence the wild-type (WT) copy of the MeCP2 gene. Thus, reactivating the silent WT copy of MeCP2 could provide therapeutic intervention for RTT. METHODS: Toward this goal, we screened ~ 28,000 small-molecule compounds from several libraries using a MeCP2-luciferase reporter cell line and cortical neurons from a MeCP2-EGFP mouse model. We used gain/increase of luminescence or fluorescence as a readout of MeCP2 reactivation and tested the efficacy of these drugs under different drug regimens, conditions, and cellular contexts. RESULTS: We identified inhibitors of the JAK/STAT pathway as XCI-reactivating agents, both by in vitro and ex vivo assays. In particular, we show that AG-490, a Janus Kinase 2 (JAK2) kinase inhibitor, and Jaki, a pan JAK/STAT inhibitor, are capable of reactivating MeCP2 from the inactive X chromosome, in different cellular contexts. CONCLUSIONS: Our results suggest that inhibition of the JAK/STAT pathway is a new potential pathway to reinstate MeCP2 gene expression as an efficient RTT treatment.

A Comparison of Three Child OHRQoL Measures.

Comparing oral health-related quality of life (OHRQoL) measures can facilitate selecting the most appropriate one for a particular research question/setting. Three child OHRQoL measures Child Perceptions Questionnaire (CPQ11⁻14), the Child Oral Health Impact Profile (COHIP) and the Caries Impacts and Experiences Questionnaire for Children (CARIES-QC) were used with 335 10- to 13-year-old participants in a supervised tooth-brushing programme in New Zealand. The use of global questions enabled their validity to be examined. Assessments were conducted at baseline and after 12 months. All three measures had acceptable internal consistency reliability. There were moderate, positive correlations among their scores, and all showed differences in the impact of dental caries on OHRQoL, with children with the highest caries experience having the highest scale scores. Effect sizes were used to assess meaningful change. The CPQ11⁻14 and the CARIES-QC showed meaningful change. The COHIP-SF score showed no meaningful change. Among children reporting improved OHRQoL, baseline and follow-up scores differed significantly for the CPQ11⁻14 and CARIES-QC measures, although not for the COHIP-SF. The three scales were broadly similar in their conceptual basis, reliability and validity, but responsiveness of the COHIP-SF was questionable, and the need to compute two different scores for the CARIES-QC meant that its administrative burden was considerably greater than for the other two measures. Replication and use of alternative approaches to measuring meaningful change are suggested.

Ube3a reinstatement mitigates epileptogenesis in Angelman syndrome model mice.

Angelman syndrome (AS) is a neurodevelopmental disorder in which epilepsy is common (~90%) and often refractory to antiepileptics. AS is caused by mutation of the maternal allele encoding the ubiquitin protein ligase E3A (UBE3A), but it is unclear how this genetic insult confers vulnerability to seizure development and progression (i.e., epileptogenesis). Here, we implemented the flurothyl kindling and retest paradigm in AS model mice to assess epileptogenesis and to gain mechanistic insights owed to loss of maternal Ube3a. AS model mice kindled similarly to wild-type mice, but they displayed a markedly increased sensitivity to flurothyl-, kainic acid-, and hyperthermia-induced seizures measured a month later during retest. Pathological characterization revealed enhanced deposition of perineuronal nets in the dentate gyrus of the hippocampus of AS mice in the absence of overt neuronal loss or mossy fiber sprouting. This pro-epileptogenic phenotype resulted from Ube3a deletion in GABAergic but not glutamatergic neurons, and it was rescued by pancellular reinstatement of Ube3a at postnatal day 21 (P21), but not during adulthood. Our results suggest that epileptogenic susceptibility in AS patients is a consequence of the dysfunctional development of GABAergic circuits, which may be amenable to therapies leveraging juvenile reinstatement of UBE3A.

Characterization and structure-activity relationships of indenoisoquinoline-derived topoisomerase I inhibitors in unsilencing the dormant Ube3a gene associated with Angelman syndrome.

Background: Angelman syndrome (AS) is a severe neurodevelopmental disorder lacking effective therapies. AS is caused by mutations in ubiquitin protein ligase E3A (UBE3A), which is genomically imprinted such that only the maternally inherited copy is expressed in neurons. We previously demonstrated that topoisomerase I (Top1) inhibitors could successfully reactivate the dormant paternal allele of Ube3a in neurons of a mouse model of AS. We also previously showed that one such Top1 inhibitor, topotecan, could unsilence paternal UBE3A in induced pluripotent stem cell-derived neurons from individuals with AS. Although topotecan has been well-studied and is FDA-approved for cancer therapy, its limited CNS bioavailability will likely restrict the therapeutic use of topotecan in AS. The goal of this study was to identify additional Top1 inhibitors with similar efficacy as topotecan, with the expectation that these could be tested in the future for safety and CNS bioavailability to assess their potential as AS therapeutics. Methods: We tested 13 indenoisoquinoline-derived Top1 inhibitors to identify compounds that unsilence the paternal allele of Ube3a in mouse neurons. Primary cortical neurons were isolated from embryonic day 14.5 (E14.5) mice with a Ube3a-YFP fluorescent tag on the paternal allele (Ube3am+/pYFP mice) or mice that lack the maternal Ube3a allele and hence model AS (Ube3am-/p+ mice). Neurons were cultured for 7 days, treated with drug for 72 h, and examined for paternal UBE3A protein expression by Western blot or fluorescence immunostaining. Dose responses of the compounds were determined across a log range of drug treatments, and cytotoxicity was tested using a luciferase-based assay. Results: All 13 indenoisoquinoline-derived Top1 inhibitors unsilenced paternal Ube3a. Several compounds exhibited favorable paternal Ube3a unsilencing properties, similar to topotecan, and of these, indotecan (LMP400) was the most effective based on estimated Emax (maximum response of unsilencing paternal Ube3a) and EC50 (half maximal effective concentration). Conclusions: We provide pharmacological profiles of indenoisoquinoline-derived Top1 inhibitors as paternal Ube3a unsilencers. All 13 tested compounds were effective at unsilencing paternal Ube3a, although with variable efficacy and potency. Indotecan (LMP400) demonstrated a better pharmacological profile of Ube3a unsilencing compared to our previous lead compound, topotecan. Taken together, indotecan and its structural analogues are potential AS therapeutics whose translational potential in AS treatment should be further assessed.

Common Pathophysiology in Multiple Mouse Models of Pitt-Hopkins Syndrome.

Mutations or deletions of the transcription factor TCF4 are linked to Pitt-Hopkins syndrome (PTHS) and schizophrenia, suggesting that the precise pathogenic mutations dictate cellular, synaptic, and behavioral consequences. Here, we generated two novel mouse models of PTHS, one that mimics the most common pathogenic TCF4 point mutation (human R580W, mouse R579W) and one that deletes three pathogenic arginines, and explored phenotypes of these lines alongside models of pan-cellular or CNS-specific heterozygous Tcf4 disruption. We used mice of both sexes to show that impaired Tcf4 function results in consistent microcephaly, hyperactivity, reduced anxiety, and deficient spatial learning. All four PTHS mouse models demonstrated exaggerated hippocampal long-term potentiation (LTP), consistent with deficits in hippocampus-mediated behaviors. We further examined R579W mutant mice and mice with pan-cellular Tcf4 heterozygosity and found that they exhibited hippocampal NMDA receptor hyperfunction, which likely drives the enhanced LTP. Together, our data pinpoint convergent neurobiological features in PTHS mouse models and provide a foundation for preclinical studies and a rationale for testing whether NMDAR antagonists might be used to treat PTHS.SIGNIFICANCE STATEMENT Pitt-Hopkins syndrome (PTHS) is a rare neurodevelopmental disorder associated with TCF4 mutations/deletions. Despite this genetic insight, there is a need to identify the function of TCF4 in the brain. Toward this goal, we developed two mouse lines, including one harboring the most prevalent pathogenic point mutation, and compared them with two existing models that conditionally delete Tcf4 Our data identify a set of overlapping phenotypes that may serve as outcome measures for preclinical studies of PTHS treatments. We also discovered penetrant enhanced synaptic plasticity across mouse models that may be linked to increased NMDA receptor function. These data reveal convergent neurobiological characteristics of PTHS mouse models and support the further investigation of NMDA receptor antagonists as a possible PTHS treatment.

Responsiveness of 2 scales to evaluate lateropulsion or pusher syndrome recovery after stroke.

OBJECTIVES: To analyze responsiveness to change of 2 outcome measures in the lateropulsion population after stroke. Lateropulsion describes an atypical balance problem after stroke where patients actively push themselves toward their paretic side. Secondary aims were to measure the incidence of lateropulsion and evaluate the site of the brain most commonly involved in lateropulsion. DESIGN: Stroke patients were screened for lateropulsion on admission to rehabilitation. Those demonstrating lateropulsion were assessed every 2 weeks using 2 scales to measure progress. Analysis of variance and the standardized response mean (SRM) were used to analyze change for each scale. SETTING: Rehabilitation and geriatric units. PARTICIPANTS: Prospective purposive sample of stroke patients (N=43) demonstrating lateropulsion on admission from a consecutive admission sample of 160. To allow comparison, data from 43 stroke patients who did not display lateropulsion were collected retrospectively, matched on age (±5y) and sex. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Incidence of lateropulsion was calculated as a percentage of stroke patients admitted. Responsiveness to change was measured by using the Burke Lateropulsion Scale (BLS), to quantify severity of lateropulsion, and the Postural Assessment Scale for Stroke (PASS), which measures postural abilities. RESULTS: Of 160 stroke patients, 26.9% displayed lateropulsion (mean age, 72y; 51% men). The BLS and PASS had high levels of measurement responsiveness (BLS SRM =1.48 and 2.24; PASS SRM =1.76 and 1.87) at 4 and 8 weeks, respectively. CONCLUSIONS: The BLS and PASS are responsive scales to monitor progress and recovery during rehabilitation. As more than 25% of stroke patients admitted to rehabilitation may exhibit lateropulsion, these 2 scales could be valuable in monitoring progress and designing future intervention studies.