Focused Ultrasound-Mediated Disruption of the Blood-Brain Barrier for AAV9 Delivery in a Mouse Model of Huntington's Disease.

Huntington's disease (HD) is a monogenic neurodegenerative disorder caused by a cytosine-adenine-guanine (CAG) trinucleotide repeat expansion in the HTT gene. There are no cures for HD, but the genetic basis of this disorder makes gene therapy a viable approach. Adeno-associated virus (AAV)-miRNA-based therapies have been demonstrated to be effective in lowering HTT mRNA; however, the blood-brain barrier (BBB) poses a significant challenge for gene delivery to the brain. Delivery strategies include direct injections into the central nervous system, which are invasive and can result in poor diffusion of viral particles through the brain parenchyma. Focused ultrasound (FUS) is an alternative approach that can be used to non-invasively deliver AAVs by temporarily disrupting the BBB. Here, we investigate FUS-mediated delivery of a single-stranded AAV9 bearing a cDNA for GFP in 2-month-old wild-type mice and the zQ175 HD mouse model at 2-, 6-, and 12-months. FUS treatment improved AAV9 delivery for all mouse groups. The delivery efficacy was similar for all WT and HD groups, with the exception of the zQ175 12-month cohort, where we observed decreased GFP expression. Astrocytosis did not increase after FUS treatment, even within the zQ175 12-month group exhibiting higher baseline levels of GFAP expression. These findings demonstrate that FUS can be used to non-invasively deliver an AAV9-based gene therapy to targeted brain regions in a mouse model of Huntington's disease.

Nacc1 Mutation in Mice Models Rare Neurodevelopmental Disorder with Underlying Synaptic Dysfunction.

A missense mutation in the transcription repressor Nucleus accumbens-associated 1 (NACC1) gene at c.892C>T (p.Arg298Trp) on chromosome 19 causes severe neurodevelopmental delay ( Schoch et al., 2017). To model this disorder, we engineered the first mouse model with the homologous mutation (Nacc1+/R284W ) and examined mice from E17.5 to 8 months. Both genders had delayed weight gain, epileptiform discharges and altered power spectral distribution in cortical electroencephalogram, behavioral seizures, and marked hindlimb clasping; females displayed thigmotaxis in an open field. In the cortex, NACC1 long isoform, which harbors the mutation, increased from 3 to 6 months, whereas the short isoform, which is not present in humans and lacks aaR284 in mice, rose steadily from postnatal day (P) 7. Nuclear NACC1 immunoreactivity increased in cortical pyramidal neurons and parvalbumin containing interneurons but not in nuclei of astrocytes or oligodendroglia. Glial fibrillary acidic protein staining in astrocytic processes was diminished. RNA-seq of P14 mutant mice cortex revealed over 1,000 differentially expressed genes (DEGs). Glial transcripts were downregulated and synaptic genes upregulated. Top gene ontology terms from upregulated DEGs relate to postsynapse and ion channel function, while downregulated DEGs enriched for terms relating to metabolic function, mitochondria, and ribosomes. Levels of synaptic proteins were changed, but number and length of synaptic contacts were unaltered at 3 months. Homozygosity worsened some phenotypes including postnatal survival, weight gain delay, and increase in nuclear NACC1. This mouse model simulates a rare form of autism and will be indispensable for assessing pathophysiology and targets for therapeutic intervention.

Early whole-body mutant huntingtin lowering averts changes in proteins and lipids important for synapse function and white matter maintenance in the LacQ140 mouse model.

Expansion of a triplet repeat tract in exon 1 of the HTT gene causes Huntington's disease (HD). The mutant HTT protein (mHTT) has numerous aberrant interactions with diverse, pleiomorphic effects. Lowering mHTT is a promising approach to treat HD, but it is unclear when lowering should be initiated, how much is necessary, and what duration should occur to achieve benefits. Furthermore, the effects of mHTT lowering on brain lipids have not been assessed. Using a mHtt-inducible mouse model, we analyzed mHtt lowering initiated at different ages and sustained for different time-periods. mHTT protein in cytoplasmic and synaptic compartments of the striatum was reduced 38-52%; however, there was minimal lowering of mHTT in nuclear and perinuclear regions where aggregates formed at 12 months of age. Total striatal lipids were reduced in 9-month-old LacQ140 mice and preserved by mHtt lowering. Subclasses important for white matter structure and function including ceramide (Cer), sphingomyelin (SM), and monogalactosyldiacylglycerol (MGDG), contributed to the reduction in total lipids. Phosphatidylinositol (PI), phosphatidylserine (PS), and bismethyl phosphatidic acid (BisMePA) were also changed in LacQ140 mice. Levels of all subclasses except ceramide were preserved by mHtt lowering. mRNA expression profiling indicated that a transcriptional mechanism contributes to changes in myelin lipids, and some but not all changes can be prevented by mHtt lowering. Our findings suggest that early and sustained reduction in mHtt can prevent changes in levels of select striatal proteins and most lipids, but a misfolded, degradation-resistant form of mHTT hampers some benefits in the long term.

On the solution arising in two-cylinders electrostatics.

The electrostatics of two cylinders charged to the symmetrical or anti-symmetrical potential is investigated by using the null-field boundary integral equation (BIE) in conjunction with the degenerate kernel of the bipolar coordinates. The undetermined coefficient is obtained according to the Fredholm alternative theorem. The uniqueness of solution, infinite solution, and no solution are examined therein. A single cylinder (circle or ellipse) is also provided for comparison. The link to the general solution space is also done. The condition at infinity is also correspondingly examined. The flux equilibrium along circular boundaries and the infinite boundary is also checked as well as the contribution of the boundary integral (single and double layer potential) at infinity in the BIE is addressed. Ordinary and degenerate scales in the BIE are both discussed. Furthermore, the solution space represented by the BIE is explained after comparing it with the general solution. The present finding is compared to those of Darevski[2] and Lekner[4] for identity.

Early whole-body mutant huntingtin lowering averts changes in proteins and lipids important for synapse function and white matter maintenance in the LacQ140 mouse model.

Expansion of a triplet repeat tract in exon1 of the HTT gene causes Huntington's disease (HD). The mutant HTT protein (mHTT) has numerous aberrant interactions with diverse, pleiomorphic effects. No disease modifying treatments exist but lowering mutant huntingtin (mHTT) by gene therapy is a promising approach to treat Huntington's disease (HD). It is not clear when lowering should be initiated, how much lowering is necessary and for what duration lowering should occur to achieve benefits. Furthermore, the effects of mHTT lowering on brain lipids have not been assessed. Using a mHtt-inducible mouse model we analyzed whole body mHtt lowering initiated at different ages and sustained for different time-periods. Subcellular fractionation (density gradient ultracentrifugation), protein chemistry (gel filtration, western blot, and capillary electrophoresis immunoassay), liquid chromatography and mass spectrometry of lipids, and bioinformatic approaches were used to test effects of mHTT transcriptional lowering. mHTT protein in cytoplasmic and synaptic compartments of the caudate putamen, which is most affected in HD, was reduced 38-52%. Little or no lowering of mHTT occurred in nuclear and perinuclear regions where aggregates formed at 12 months of age. mHtt transcript repression partially or fully preserved select striatal proteins (SCN4B, PDE10A). Total lipids in striatum were reduced in LacQ140 mice at 9 months and preserved by early partial mHtt lowering. The reduction in total lipids was due in part to reductions in subclasses of ceramide (Cer), sphingomyelin (SM), and monogalactosyldiacylglycerol (MGDG), which are known to be important for white matter structure and function. Lipid subclasses phosphatidylinositol (PI), phosphatidylserine (PS), and bismethyl phosphatidic acid (BisMePA) were also changed in LacQ140 mice. Levels of all subclasses other than ceramide were preserved by early mHtt lowering. Pathway enrichment analysis of RNAseq data imply a transcriptional mechanism is responsible in part for changes in myelin lipids, and some but not all changes can be rescued by mHTT lowering. Our findings suggest that early and sustained reduction in mHtt can prevent changes in levels of select striatal proteins and most lipids but a misfolded, degradation-resistant form of mHTT hampers some benefits in the long term.

Chinese validation of Urogenital Distress Inventory and Incontinence Impact Questionnaire short form.

INTRODUCTION AND HYPOTHESIS: UDI-6 and IIQ-7 are useful disease-specific questionnaires evaluating the impact of urinary incontinence on the QOL of women. We aim at validating them in Chinese language. METHODS: Both instruments were translated; 207 urinary incontinent women completed UDI-6 and IIQ-7, SF-36, bladder diary and urodynamic evaluation. The reliability and validity were assessed. RESULTS: There were high internal consistency (Cronbach's alpha for UDI-6 and IIQ-7 was 0.80 and 0.93) and test-retest reliability (Intraclass correlation coefficient was 0.72 and 0.75, P < 0.001). Scoring of UDI-6 and IIQ-7 was negatively correlated with SF-36 (P < 0.001); positively correlated with daytime urinary frequency and incontinent episodes (P < 0.001), and women's VAS (P < 0.001). Subscales of UDI-6 and IIQ-7 could discriminate women with different urodynamic diagnoses. CONCLUSIONS: The Chinese UDI-6 and IIQ-7 is reliable and valid. Study on the responsiveness to treatment is in progress. They are useful in assessing impact of the urinary incontinence in Chinese women.

A 1-year longitudinal study of psychological morbidity after miscarriage.

OBJECTIVE: To examine the 1-year longitudinal course of psychological outcomes after miscarriage. DESIGN: Longitudinal observational study. SETTING: University-affiliated teaching hospital. PATIENT(S): 280 miscarrying women and 150 nonpregnant women. INTERVENTION(S): Psychological outcomes were assessed using the 12-item General Health Questionnaire (GHQ-12) and Beck Depression Inventory (BDI) immediately, 3 months, 6 months, and 12 months after miscarriage. MAIN OUTCOME MEASURE(S): Scores on GHQ-12 and BDI. RESULT(S): Half (55%) of the miscarrying women scored high (>or=4) on the GHQ-12 immediately, 25% at 3 months; 17.8% at 6 months, and 10.8% at 1 year after miscarriage; 26.8% of the patients scored high on the BDI (>or=12) immediately, 18.4% at 3 months, 16.4% at 6 months, and 9.3% at 1 year after miscarriage. Patients who were initially more distressed continued to score higher on both the GHQ-12 and the BDI along the 1-year course when compared with those who were initially less distressed. When compared with the nonpregnant controls, the miscarrying women scored statistically significantly higher on the GHQ-12 and BDI; the differences became not statistically significant only 1 year after miscarriage. CONCLUSION(S): A statistically significant proportion of patients reported psychological morbidity shortly after miscarriage, but their level of distress reduced over time until they were comparable with the controls 1 year later. Patients who were initially more distressed continued to be throughout the 1-year course.

Prevalence and associated risk factors of retention of urine after caesarean section.

Caesarean section poses higher risk of postpartum urinary retention (PUR) than vaginal delivery. The aim of this study was to identify the prevalence and the associated risk factors of PUR after caesarean section. Two hundred seven parturients (mean age = 31.26 years old, median parity = 0) who delivered by caesarean section were recruited from the postnatal unit of a tertiary hospital in Hong Kong. Voiding was encouraged 6 h after removal of Foley catheters. Transvaginal ultrasound scan was performed immediately after voiding to estimate the postvoid residual bladder volume (PVR). PUR after caesarean section was defined as PVR of more than 150 ml. The results indicated a prevalence of 3.38%. Logistic regression analysis indicating 'lack of progress of labor' was the only significant associated factor (p < 0.001). The findings of this study provide information for further exploration on how to reduce the morbidity caused by PUR in the postoperative period of caesarean section.

Psychological morbidity and female urinary incontinence.

Female urinary incontinence is a common yet distressing condition. It affects women of all ages, but is especially common in the elderly. Its prevalence ranges from 15 to 55%, depending on age and population studied. Despite the ubiquity of female urinary incontinence, many incontinent women do not voice their suffering, and urinary incontinence has therefore been dubbed 'the silent epidemic'. The physical impact and social isolation associated with urinary incontinence lead to impairment of quality of life and psychological well-being. It is the aim of this review to discuss the epidemiology and psychological impact of urinary incontinence. The recognition and management of incontinence-related psychological morbidity are also explored.

Measurement of tension-free vaginal tape trocar insertion and exit forces.

BACKGROUND/AIMS: This study aims to establish a model to measure the forces required for tension-free vaginal tape (TVT) trocar application using a pressure transducer. METHODS: A 3.5 x 3.5 cm pressure transducer was constructed from piezo-resistive material. The transducer digitally recorded the surface contact pressure (pressure = force/area) profile continuously during TVT trocar application. Recordings were successfully obtained from 24 subjects. RESULTS: The mean (standard error) peak contact pressures during application at the right and left side vaginal walls, and right and left side abdominal walls were 1.05 (0.12) pounds per square inch (psi), 1.08 (0.21) psi, 2.49 (0.3) psi and 2.49 (0.3) psi, respectively. On both the right and left side, pair-wise comparisons between trocar insertion and exit forces were significantly different (p < 0.001). CONCLUSION: A model has been developed by which TVT trocar application forces can be objectively measured. The TVT trocar exit force is substantially greater than the insertion force, regardless of side.

Changing prevalence and knowledge of urinary incontinence among Hong Kong Chinese women.

A territory-wide telephone survey was conducted in Hong Kong to assess the prevalence, knowledge, and treatment-seeking behaviour of Chinese women with urinary incontinence, using validated Chinese version of Urogenital Distress Inventory (UDI-6) and Incontinence Impact Questionnaire (IIQ-7). Women, 540, aged between 17 to 77 years were interviewed. Of the respondents, 40.8% reported stress urinary incontinence, 20.4% had urge incontinence and 15.9% had mixed incontinence. Among these, 16.0% reported quality of life impairment; 9.3% felt frustrated with low morale, and 15.2% had nervous and anxiety problems. However, as many as 78.3% of the respondents did not know that stress urinary incontinence is a disease entity, and 60.6% thought that leakage of urine was a normal aging process. For those respondents having stress urinary incontinence, the first treatment of choice was physiotherapy. The second choice was medication, and surgical treatment was the last option. Respondents with stress urinary incontinence showed higher education level.

Long-term reproductive outcome subsequent to medical versus surgical treatment for miscarriage.

BACKGROUND: When compared with the conventional surgical evacuation for the treatment of miscarriage, medical evacuation has been largely accepted as an effective and safe management. However, there is a lack of data on the long-term reproductive outcome of these two treatment modalities, which is crucial in patient counselling. The current study evaluates and compares the long-term fertility and pregnancy outcome following these two treatments. METHODS: A cohort of 604 women enrolled in a previous randomized controlled trial comparing medical and surgical evacuation for miscarriage were followed up prospectively by telephone interview at a median of 6 (range 4-9) years using a structured questionnaire. RESULTS: A total of 423 women were contacted and four declined to participate (response rate 69.4%). Of these, 261 women (131 medical and 130 surgical evacuations) had attempted to become pregnant since the miscarriage. There were no differences in their baseline characteristics including age, reproductive and contraceptive history. The natural conception rates were the same (97.7%, P = 0.99) and the cumulative pregnancy rates were similar between groups, being 60 and 80% at 12 and 24 months respectively. The median time-to-pregnancy was 8 months in both groups (P = 0.97) and the subsequent live birth rates (85.2 versus 88.2%, P = 0.72) resulting from the immediate pregnancy following previous treatment were similar. CONCLUSIONS: The long-term conception rate and pregnancy outcome are not different following medical or surgical evacuation for miscarriage. Women should be reassured that their long-term fertility potential will not be compromised after medical treatment.

Screening test model using duration of labor for the detection of postpartum urinary retention.

AIMS: To construct a screening test model for postpartum urinary retention (PUR), using the duration of labor, which was shown to be the risk factor for PUR. METHODS: A group of 691 consecutive patients were recruited at postpartum day 1. Of the 691 patients, 101 (14.6%) had PUR. The 691 patients were computer-randomized into two groups, 'A' and 'B.' A receiver operating characteristic (ROC) curve was constructed for Group A to determine the optimum cutoff value for screening PUR using the duration of labor. The cutoff value was then applied to Group B to determine the screening test characteristics of the duration of labor: sensitivity, specificity, predictive values, and likelihood ratios. RESULTS: An ROC curve with an area under curve (AUC) of 0.63 (95% CI 0.57-0.69, P < 0.0001) was constructed. A range of cutoff values (with specificity ranging from 0.60 to 0.99) for the duration of labor was determined and the test characteristics computed. According to the test characteristics the optimal cutoff value for the duration of labor was 700 min. This cutoff value for the duration of labor has a specificity of 0.95, negative predicative value of 0.86, and likelihood ratio for a positive test of 0.88. CONCLUSION: A screening test model using the duration of labor can be constructed to predict PUR.

Screening for post-miscarriage psychiatric morbidity.

OBJECTIVE: The purpose of this study was to evaluate 12-item General Health Questionnaire (GHQ-12) in screening for psychiatric morbidity after miscarriage. STUDY DESIGN: A prospective cohort study was carried out involving 222 patients. Six weeks after miscarriage, the GHQ-12 was applied. Psychiatric "case" or "non-case" was diagnosed by the psychiatrist with use of the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders-III-R. The patients were computer randomized into Groups A or B. A receiver operating characteristic (ROC) curve was constructed for Group A. The optimal cutoff value of GHQ-12 was determined, and this value was applied to Group B. The test characteristics were assessed. RESULTS: Twenty-seven patients were found to be psychiatric cases. An ROC with area under curve of 0.93 (95% CI 0.87-0.99, P<.001) was constructed. The best GHQ-12 cutoff score was > or =4 in detecting psychiatric caseness. A sensitivity of 83%, specificity of 90%, positive predictive value of 50%, and negative predictive value of 98% were obtained. CONCLUSION: GHQ-12 is an effective screening tool in detecting psychiatric morbidity after miscarriage.

A study of female urinary tract infection caused by urodynamic investigation.

OBJECTIVE: The purpose of this study was to assess the prevalence of female urinary tract infection before and after urodynamic investigation and to identify the risk factors for urinary tract infection after urodynamic investigation. STUDY DESIGN: Eight hundred twenty-two consecutive incontinent women were recruited. All women were "double-screened" and treated for urinary tract infection before urodynamic investigation: first by mid stream urine culture 4 to 6 weeks before investigation and then by reagent strips for urine leukocytes and nitrites at the time of investigation. The investigation was postponed until the urinary tract infection had been treated. All women then received a standard urodynamic investigation. RESULTS: The prevalence of urinary tract infection before urodynamic investigation was 5.1% (95% CI, 3.6-6.6), and the prevalence after the investigation was 8.4% (95% CI, 6.5-10.3). Three independent risk factors were identified: age >or=70 years (odds ratio, 1.99; 95% CI, 1.14-3.48), previous continence surgery (odds ratio, 1.90; 95% CI, 1.05-3.43), and urinary tract infection before urodynamic investigation (odds ratio, 3.13; 95% CI, 1.43-6.83). The 3 most common uropathogens in the urinary tract infections after the urodynamic investigation were Escherichia coli (46.3%), Enterococcus spp (16.4%), and Enterococcus faecalis (11.9%). CONCLUSION: Despite a stringent screen-and-treat protocol before urodynamic investigation, patients still experienced urinary tract infection.