Microglial expression of GAT-1 in the cerebral cortex.

Microglial cells are the immune cells of the brain that, by sensing the microenvironment, permit a correct brain development and function. They communicate with other glial cells and with neurons, releasing and responding to a number of molecules that exert effects on surrounding cells. Among these, neurotransmitters and, in particular, gamma-aminobutyric acid (GABA) has recently gained interest in this context. We demonstrated the expression of GABA transporter 1 (GAT-1) in microglial cells both in soma and cell processes. We show that microglial cell treatment with 1,2,5,6-tetrahydro-1-[2-[[(diphenylmethylene)amino]oxy]ethyl]-3-pyridinecarboxylic acid hydrochloride (NNC-711), a potent and selective GAT-1 inhibitor, significantly reduced Na+ -dependent GABA uptake. On the other hand, GABA uptake was significantly increased by cell treatment with (S)-1-[2-[tris(4-methoxyphenyl)methoxy]ethyl]-3-piperidinecarboxylic acid (SNAP-5114), a GAT-2/3 inhibitor, and this effect was completely blocked by the botulinum toxin BoNT/C1, that specifically cleaves and inactives syntaxin 1A (STX1A). Overall, these findings show that microglial cells express GAT-1 and indicate that STX1A plays an important role in the regulation of GAT-1-dependent GABA uptake in microglia.

Robot-assisted laparoscopic artificial urinary sphincter insertion in women with stress urinary incontinence: a pilot single-centre study.

OBJECTIVE: To report the functional outcomes of robot-assisted laparoscopic artificial urinary sphincter (AUS; AMS 800™, American Medical Systems, Minnetonka, MN, USA) implantation and revision in women with stress urinary incontinence (SUI). PATIENTS AND METHODS: A pilot prospective monocentric study included all consecutive female patients with SUI and undergoing an AUS surgery (implantation or revision) using a robotic approach between 2012 and 2018. The AUS was implanted through a transperitoneal robotic approach. The dissection of the bladder neck was initiated using a posterior approach with a permanent visual control. Intraoperative and early postoperative complications were reported (Clavien-Dindo classification). Continence was defined as no pad usage. RESULTS: A total of 41 patients, median (interquartile range [IQR]) age 67.5 (57-74.7) years were included. In the group of 27 patients undergoing an AUS implantation, one conversion to laparotomy, one bladder neck and one ureteric injury occurred. The AUS was not implanted in the two latest cases. Twelve minor (Clavien-Dindo Grade I-II) early postoperative complications occurring in 10 patients were reported. The median (IQR) follow-up was 19 (11-27) months. The continence rate was 84% (21/25 patients). In the group of 14 patients undergoing an AUS revision, no vaginal or bladder injury was reported. One patient had a bowel injury with conversion to laparotomy without AUS insertion. Two major (Clavien-Dindo Grade III-V) early postoperative complications occurred in two patients: one intraoperative bowel injury, which ultimately resulted in the death of the patient and one device infection requiring an explantation of the AUS. The median (IQR) follow-up was 18 (13.5-24.2) months. The continence rate was 83.3% (10/12 patients). CONCLUSION: The results obtained after robot-assisted laparoscopic AUS implantation among women are promising despite the significant morbidity due to previous pelvic surgeries. Longer follow-up studies are needed.

Pure stress urinary incontinence: analysis of prevalence, estimation of costs, and financial impact.

BACKGROUND: The prevalence of pure stress urinary incontinence (P-SUI) and the role of urodynamic investigation (UDI) prior to surgery for stress urinary incontinence (SUI) is debated. Since the exact prevalence of P-SUI is not clear, its clinical and economic impact is not well defined. The aims of this study were to evaluate the prevalence of P-SUI in a population of women who underwent UDI for urinary incontinence (UI), also assessing: 1) the correspondence between clinical diagnosis of P-SUI and urodynamic findings; 2) the analysis of costs in terms of UDI and eventually post-UDI avoided surgical procedures. METHODS: A single cohort of women who underwent UDI for UI between January 2012 and July 2016 was prospectively collected and retrospectively analyzed. Clinical P-SUI was defined by the strict criteria of the International Continence Society. For each patient, history, physical examination and UDI were collected. The correspondence between clinical and urodynamic findings of P-SUI was analyzed. The rate of clinical P-SUI changed after performing UDI and the number of unnecessary intervention after UDI were reported. A wide cost analysis of UDIs, and the amount of surgical procedures that were believed unnecessary after UDI was reported. RESULTS: Stress urinary incontinence was present in 323/544 (59.4%) patients. The prevalence of clinical P-SUI was 20.7% (67/323), while the prevalence of complicated SUI (C-SUI) was 79.3% (256/323). After UDI, diagnosis of P-SUI decreased to 18.3% (59/232). In 10.2% of cases (6/59) the scheduled middle urethral sling (MUS) was suppressed after the UDI results because 3/6 cases had detrusor overactivity and urge incontinence, in 2/6 cases SUI was treated with a conservative management, in 1/6 case an important voiding dysfunction was detected. Considering the national reimbursement in our country, the cost of each UDI was 296.5 euros and the total amount was 17,493.5 euros. So far the surgery-related savings covered 61.7-105.0% of the costs of total number of UDIs performed in the uncomplicated patients. CONCLUSIONS: The prevalence of clinical P-SUI is relevant, involving about 20% of women with clinical SUI. Although the correspondence between clinical and urodynamic diagnosis was high, we demonstrated that UDI may help in some cases to avoid an inappropriate surgical treatment. Therefore, UDI prior to SUI surgery should be considered to achieve a correct diagnosis and a proper therapeutic strategy.

Health-related quality of life after BCG or MMC induction for non-muscle invasive bladder cancer.

INTRODUCTION: To evaluate health-related quality of life (HRQoL) in patients with non-muscle invasive bladder cancer (NMIBC) during the induction phase of intravesical instillations with BCG or MMC. MATERIALS AND METHODS: HRQoL was measured by two questionnaires from EORTC (QLQ-C30 and QLQ-BLS24), stratifying results by gender, age and therapy at the start of the therapy (T0), at last instillation (T1) and at 3 months after T1 (T2). The persistence of QoL-related side effects after 3 months from the end of the induction cycle was evaluated. RESULTS: We enrolled 108 naïve patients and 103 patients self-completed the questionnaires. Treatment was well tolerated in both groups. Side effects were reported by 46.6% of patients at T1 and 47.5% of patients at T2. QoL dropped at T1, returning to the baseline at T2. Drop of QoL was greater in the physical, role, emotional and social functioning domains and in some clinical domains as pain, fatigue and insomnia. Our stratified analysis showed that patients > 70 years have a worsening of QoL, a higher incidence of patient-reported side effects or symptoms in the BCG arm as compared to MMC arm. CONCLUSIONS: Our study shows that intravesical instillations of BCG or MMC during the induction phase might have a relevant effect on HRQoL.

GAT-1 mediated GABA uptake in rat oligodendrocytes.

Stimulated by the results of a recent paper on the effects of tiagabine, a selective inhibitor of the main GABA transporter GAT-1, on oligodendrogenesis, we verified the possibility that GAT-1 may be expressed in oligodendrocytes using immunocytochemical methods and functional assays. Light microscopic analysis of the subcortical white matter of all animals revealed the presence of numerous GAT-1+ cells of different size (from 3 to 29 µm) and morphology. An electron microscope analysis revealed that, besides fibrous astrocytes and interstitial neurons, GAT-1 immunoreactivity was present in immature and mature oligodendrocytes. Co-localization studies between GAT-1 and markers specific for oligodendrocytes (NG2 and RIP) showed that about 12% of GAT-1 positive cells in the white matter were immature oligodendrocytes, while about 15% were mature oligodendrocytes. In vitro functional assays showed that oligodendrocytes exhibit tiagabine-sensitive Na+ -dependent GABA uptake. Although relationships between GABA and oligodendrocytes have been known for many years, this is the first demonstration that GAT-1 is expressed in oligodendrocytes. The present results on the one hand definitely closes the era of "neuronal" and "glial" GABA transporters, on the other they suggest that oligodendrocytes may contribute to pathophysiology of the several diseases in which GAT-1 have been implicated to date. GLIA 2017;65:514-522.

Epigenetics of Huntington's Disease.

Huntington's disease (HD) is a genetic, fatal autosomal dominant neurodegenerative disorder typically occurring in midlife with symptoms ranging from chorea, to dementia, to personality disturbances (Philos Trans R Soc Lond Ser B Biol Sci 354:957-961, 1999). HD is inherited in a dominant fashion, and the underlying mutation in all cases is a CAG trinucleotide repeat expansion within exon 1 of the HD gene (Cell 72:971-983, 1993). The expanded CAG repeat, translated into a lengthened glutamine tract at the amino terminus of the huntingtin protein, affects its structural properties and functional activities. The effects are pleiotropic, as huntingtin is broadly expressed in different cellular compartments (i.e., cytosol, nucleus, mitochondria) as well as in all cell types of the body at all developmental stages, such that HD pathogenesis likely starts at conception and is a lifelong process (Front Neurosci 9:509, 2015). The rate-limiting mechanism(s) of neurodegeneration in HD still remains elusive: many different processes are commonly disrupted in HD cell lines and animal models, as well as in HD patient cells (Eur J Neurosci 27:2803-2820, 2008); however, epigenetic-chromatin deregulation, as determined by the analysis of DNA methylation, histone modifications, and noncoding RNAs, has now become a prevailing feature. Thus, the overarching goal of this chapter is to discuss the current status of the literature, reviewing how an aberrant epigenetic landscape can contribute to altered gene expression and neuronal dysfunction in HD.

Consulting "Dr. Google" for Prostate Cancer Treatment Options: A Contemporary Worldwide Trend Analysis.

BACKGROUND: In the era of digital data, the Internet has become the primary source from which individuals draw healthcare information. OBJECTIVE: The aim of the present study is to determine worldwide public interest in prostate cancer (PCa) treatments, their penetrance and variation, and how they compare over time. DESIGN, SETTING, AND PARTICIPANTS: An analysis of worldwide search-engine trends included electronic Google queries from people who searched PCa treatment options from January 2004 to August 2018, worldwide. Join-point regression was performed. Comparisons of annual relative search volume (ARSV), average annual percentage change (AAPC), and temporal patterns were analysed to assess loss or gain of interest. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Evaluations were made regarding (1) interest in PCa treatments, (2) comparison of people's interest, and (3) impact of the US Preventive Service Task Force (USPSTF) screening recommendation and National Comprehensive Cancer Network (NCCN) guideline endorsements on Internet searching for PCa treatments. RESULTS AND LIMITATIONS: The mean ARSV for "prostatectomy" was 73% in 2004 and decreased thereafter, reaching a nadir of 36% in 2014 (APC: -7.2%; 95% confidence interval [CI] -7.8, -6.7; p < 0.01). Similarly, decreased interest was recorded for radiation therapy (AAPC: -3.2%; p = 0.1), high-intensity focused ultrasound (AAPC: -2.3%; p = 0.1), hormonal therapy (AAPC: -11.6%; p < 0.01), ablation therapy (AAPC: -4.1%; p < 0.01), cryotherapy (AAPC: -9.9%; p < 0.01), and brachytherapy (AAPC: -8.3%; p < 0.01). A steep interest was found in active surveillance (AS) (AAPC: +14.2%; p < 0.01) and focal therapy (AAPC: +27.5%; p < 0.01). When trends were compared before and after NCCN and USPSTF recommendations, a consistent decrease of all the treatment options was found, while interest in focal therapy and AS showed an augmented mean ARSV (+19.6 and +31.6, respectively). CONCLUSIONS: People are increasingly searching the Internet for PCa treatment options. A parallel decrease of interest was found for the nonmonitoring treatments, except for focal therapy, while an important growth of appeal has been recorded for AS. Understanding people inquisitiveness together with their degree of knowledge could be supportive to guiding counselling in the decision-making process and putting effort in certifying patient information. PATIENT SUMMARY: In the era of digital data, patients are increasingly searching the Internet for prostate cancer (PCa) treatment options. To safeguard patients' knowledge, it is mandatory to understand how people seek healthcare information, guaranteeing certified and evidence-based information pertaining to PCa treatments options.

Multiple stones in neobladder: Case report and literature review.

INTRODUCTION: Neobladder urolithiasis is a rare but important long-term complication of orthotopic urinary diversion. It may be asymptomatic and can be discovered as an incidental finding on a radiological investigation. However, when symptoms occur, they may include lower abdominal pain, dysuria, hematuria, and lower urinary tract symptoms. CASE DESCRIPTION: We report the case of a 63-year-old male patient with irritative lower urinary tract symptoms, lower abdominal fullness, urinary incontinence, fecaluria, and urinary loss from the left inguinal fold 12 years after a radical cystoprostatectomy with a orthotopic neobladder. Computed tomography scan and urethrocystography showed a distended pouch with multiple large stones, an enterovesical fistula, and neovesicocutaneous fistula. The fistulae were successfully managed conservatively with the placement of a Foley catheter. After 3 months, open cystolithotomy was performed and approximately 50 stones with dimensions varying from 5 mm to 5 cm, with a total weight of 890 g, were removed. After a 1-year follow-up, the patient did not report pain, urinary tract infections, or symptoms suggestive of fistula and imaging evaluation confirmed no recurrence of neobladder stones. CONCLUSION: Neobladder stones may present with various symptoms. Our patient had irritative lower urinary tract symptoms, lower abdominal fullness, urinary incontinence, fecaluria, and urinary loss from the left inguinal fold 12 years after a radical cystoprostatectomy with a orthotopic neobladder. Our experience demonstrates that open cystolithotomy is an effective intervention for the removal of large stones in neobladder.

OnabotulinumtoxinA detrusor injection improves female sexual function in women with overactive bladder wet syndrome.

OBJECTIVES: The correlation between changes in sexual function and improvements in LUTD in patients treated with OnabotulinumtoxinA (onaBoNT-A) detrusor injection is unclear and limited only to women with neurogenic OAB. The aim of this study was to evaluate the impact of OnabotulinumtoxinA (onaBoNT-A) injection on sexual function in women undergoing this treatment for idiopathic wet overactive bladder (OAB). STUDY DESIGN: This is a pilot three-center observational study including women affected by idiopathic wet overactive bladder refractory to standard conservative treatments and underwent onaBoNT-A injection. Sexuality was assessed using the Female Sexual Function Index (FSFI). A 3-day voiding diary, OAB screener questionnaire (OAB-S), and the international consultation on incontinence questionnaire short form (ICIQ-sf) were completed before and 3 months after onaBoNT-A injection to evaluate OAB symptoms. RESULTS: All the 32 enrolled patients were included for statistical analysis. These patients received 100U of onaBoNT-A. Significant improvement of many FSFI domains was found. Only desire and pain domains had no significant improvements. The FSFI total score showed a significant improvement (P 0.0008). Clinical efficacy has been documented by voiding diaries, OAB-S scores, and ICIQ-sf scores. Correlation between UUI episodes and FSFI total score was statistically significant (r = -0.73; p = 0.04) while no significant correlation was found between the number of micturition and FSFI total score. CONCLUSION: Women who underwent successful OnaBoNT-A detrusor injection to treat wet OAB, showed an improvement in sexual function due to the significant correlation between the improvement of urinary urge incontinence and a better gratification of sexuality.

Differential expression of metabotropic glutamate and GABA receptors at neocortical glutamatergic and GABAergic axon terminals.

Metabotropic glutamate (Glu) receptors (mGluRs) and GABAB receptors are highly expressed at presynaptic sites. To verify the possibility that the two classes of metabotropic receptors contribute to axon terminals heterogeneity, we studied the localization of mGluR1α, mGluR5, mGluR2/3, mGluR7, and GABAB1 in VGLUT1-, VGLUT2-, and VGAT- positive terminals in the cerebral cortex of adult rats. VGLUT1-positive puncta expressed mGluR1α (∼5%), mGluR5 (∼6%), mGluR2/3 (∼22%), mGluR7 (∼17%), and GABAB1 (∼40%); VGLUT2-positive terminals expressed mGluR1α (∼10%), mGluR5 (∼11%), mGluR2/3 (∼20%), mGluR7 (∼28%), and GABAB1 (∼25%); whereas VGAT-positive puncta expressed mGluR1α (∼27%), mGluR5 (∼24%), mGluR2/3 (∼38%), mGluR7 (∼31%), and GABAB1 (∼19%). Control experiments ruled out the possibility that postsynaptic mGluRs and GABAB1 might have significantly biased our results. We also performed functional assays in synaptosomal preparations, and showed that all agonists modify Glu and GABA levels, which return to baseline upon exposure to antagonists. Overall, these findings indicate that mGluR1α, mGluR5, mGluR2/3, mGluR7, and GABAB1 expression differ significantly between glutamatergic and GABAergic axon terminals, and that the robust expression of heteroreceptors may contribute to the homeostatic regulation of the balance between excitation and inhibition.

Relationship between hepatic haemodynamics assessed by Doppler ultrasound and liver stiffness.

AIM: We tested the relationship between hepatic haemodynamics assessed by Doppler ultrasonography and liver stiffness assessed by Transient Elastography in hepatitis C related chronic liver disease. METHODS: Three liver Doppler ultrasound parameters (hepatic artery resistance index, splenic artery resistance index and waveform pattern in hepatic veins) and liver stiffness measured by Transient Elastography were analysed in one hundred consecutive patients affected by hepatitis C related chronic liver disease. RESULTS: Hepatic and splenic arteries resistance indexes correlate significantly (p<0.0001 for both) with liver stiffness. A hepatic artery resistance index cut-off value of 0.64 provided sensitivity and specificity respectively of 84.4% and 69.1% for predicting liver stiffness ≤or >13 kPa, whereas a splenic artery resistance index cut-off value of 0.56 provided sensitivity and specificity respectively of 81.3% and 48.5%. The coincidental finding of both resistance indexes above the respective cut-off values showed a good accuracy in identifying patients with liver stiffness values >13 kPa (accuracy=78%, +LR=2.90, -LR=0.31). A significant difference in liver stiffness values was evident between patients with triphasic and bi- or monophasic waveform pattern (p=0.005). CONCLUSIONS: Hepatic and splenic arteries resistance indexes and the hepatic veins waveform pattern assessed by Doppler ultrasound may provide information similar to that of Transient Elastography in hepatitis C related chronic liver disease.

Systemic and regional hemodynamics in pre-ascitic cirrhosis: effects of posture.

BACKGROUND/AIMS: To clarify the hemodynamic pattern of pre-ascitic cirrhosis, we compared the impact of posture on systemic and regional hemodynamics of patients and healthy subjects without and with plasma volume expansion. METHODS: Cardiac index (CI), peripheral vascular resistance (PVRi), heart rate, mean arterial pressure, and the mean blood flow velocities of superior mesenteric (SMAV) and common femoral arteries were evaluated by duplex-Doppler techniques in 10 patients and 20 healthy controls after 2 h of standing and 2 h after lying down. Ten healthy controls received saline infusion (15 ml/kg body weight) when they changed their posture, and five were also evaluated after plasma volume expansion in the upright posture. RESULTS: Standing systemic and regional hemodynamics did not differ between patients and controls. After saline infusion, standing control subjects showed greater CI and SMAV than patients. Recumbency caused changes of CI, PVRi and SMAV greater in patients and controls with plasma expansion than in controls without expansion, so that supine patients and controls with expansion were indistinguishable, showing higher CI and SMAV and lower PVRi than controls without expansion. CONCLUSIONS: Systemic and regional hemodynamics of patients with pre-ascitic cirrhosis are mainly determined by blood volume expansion which is compartmentalized within the splanchnic venous bed during standing and translocates towards the central and arterial circulatory districts during recumbency.

Caracterización de sueros antirreceptores de insulina: su aplicabilidad experimental y clínica / Characterization of insulin antireceptors serum: his application experimental and clinic

Se prepararon receptores de hígado de rata por centrifugación diferencial y cromatografía en Sepharosa CL-68 (purificación aproximada 1.000veces) y se inocularon con aquellos 4 conejos. La calidad de los receptores fue controlada por marcación con ATP-Y-32P, electroforesis en poliacrilamida y autorradiografía. Se observó en curvas dosis/respuesta con insulina-125I e IGF-I125I, que el aislamiento usado no excluía el receptor de IGF-I (insulina: 36 fmoles/ml, IGF-I:3,2 fmoles/ml), por lo cual se continuó la purificación en una cromatografía de afinidad de insulina-Sepharosa y posterior marcación con 125I. El antisuero seleccionado C21 demostró una inmunoprecipitación elevada con insulina y baja con IGF-I(insulina: 62,5+4,71%, IGF-I: 24,7+3,16%). En hepatocitos aislados este antisuero no compite con la insulina por su receptor y a una dilución de 1/200 tiene poca capacidad de convertir glucosa-14C a glucógeno-14C, ya que a esa dilución asocia el 50% de los receptores presentes (0,76+0,16 ng/ml, expresado en respuesta biológica equivalente a insulina). para determinar el sitio de unión del antisuero C21 se dirigial receptor de insulina con colagenasa (que digiere específicamente la subunidad B) y se observan estas condiciones la inmunoprecipitibilidad del antisuero C21 cae dramáticamente (receptor de insulina 75,6+2,04%, receptor de insulina digerido: 14,9+1,28%). Se determinó, además, la inmonorreactividad cruzada con receptores humanos para la insulina, observándose un débil cruzamiento a alta concentración (1/75: 1,95+1,32%). El estudio demuestra que el antisueroC21 es predominantemente inmunorreactivo con el receptor de insulina, al que une por su subunidad B, induciendo una escasa respuesta biológica en hepatocitos aislados.