The epistatic interaction between the dopamine D3 receptor and dysbindin-1 modulates higher-order cognitive functions in mice and humans.

The dopamine D2 and D3 receptors are implicated in schizophrenia and its pharmacological treatments. These receptors undergo intracellular trafficking processes that are modulated by dysbindin-1 (Dys). Indeed, Dys variants alter cognitive responses to antipsychotic drugs through D2-mediated mechanisms. However, the mechanism by which Dys might selectively interfere with the D3 receptor subtype is unknown. Here, we revealed an interaction between functional genetic variants altering Dys and D3. Specifically, both in patients with schizophrenia and in genetically modified mice, concomitant reduction in D3 and Dys functionality was associated with improved executive and working memory abilities. This D3/Dys interaction produced a D2/D3 imbalance favoring increased D2 signaling in the prefrontal cortex (PFC) but not in the striatum. No epistatic effects on the clinical positive and negative syndrome scale (PANSS) scores were evident, while only marginal effects on sensorimotor gating, locomotor functions, and social behavior were observed in mice. This genetic interaction between D3 and Dys suggests the D2/D3 imbalance in the PFC as a target for patient stratification and procognitive treatments in schizophrenia.

Relationship between urodynamic findings and sexual function in multiple sclerosis patients with lower urinary tract dysfunction.

BACKGROUND AND PURPOSE: Sexual dysfunction (SD) is prevalent in multiple sclerosis (MS) patients and affects quality of life. Furthermore, lower urinary tract dysfunction (LUTD) is common in MS patients. Our aim was to determine the relationship between urodynamic findings and SD in a cohort of MS patients with LUTD. METHODS: From January 2011 to September 2013, 135 consecutive patients with MS in remission phase and LUTD underwent a first urodynamic examination, according to the International Continence Society criteria. Neurological impairment was assessed using the Expanded Disability Status Scale and SD was investigated with the Female Sexual Function Index (FSFI) or the International Index of Erectile Function (IIEF-15). Multivariate logistic regression analysis was performed to identify predictors of female SD (FSFI < 26.55) or moderate-severe erectile dysfunction (ED) (IIEF-EF ≤ 16), after adjusting for confounding factors including urodynamic findings. RESULTS: Subjects with maximum detrusor pressure during involuntary detrusor contraction (PdetmaxIDC) ≥20.0 cmH2 O had lower IIEF-EF, IIEF overall satisfaction (IIEF-OS), FSFI-Arousal, FSFI-Lubrication and FSFI-Orgasm. Subjects with maximum cystometric capacity (MCC) ≥135 ml had higher IIEF-EF, intercourse satisfaction (IIEF-IS), orgasmic function (IIEF-OF), sexual desire (IIEF-SD), FSFI-Arousal, FSFI-Lubrication, FSFI-Orgasm, FSFI-Satisfaction and FSFI-Pain. On multivariate logistic regression analysis, PdetmaxIDC ≥20 cmH2 O [odds ratio (OR) 6.7; P < 0.05] and MCC <135 ml (OR 6.80; P < 0.05) were predictors of moderate-severe ED. In a model including all previous variables, compliance ≤3 ml/cmH2 O was an independent predictor of moderate-severe ED (OR 14.49; P < 0.01). No relationship was found between the previous variables and FSFI <26.55. CONCLUSIONS: Neurogenic bladder is associated with SD in MS patients. The presence of PdetmaxIDC ≥20 cmH2 O, MCC <135 ml and compliance ≤3 ml/cmH2 O may significantly predict the presence of moderate-severe ED.

Hydrophobic anions potently and uncompetitively antagonize GABA(A) receptor function in the absence of a conventional binding site.

BACKGROUND AND PURPOSE: A 'lock-and-key' binding site typically accounts for the effect of receptor antagonists. However, sulphated neurosteroids are potent non-competitive antagonists of GABA(A) receptors without a clear structure-activity relationship. To gain new insights, we tested two structurally unrelated hydrophobic anions with superficially similar properties to sulphated neurosteroids. EXPERIMENTAL APPROACH: We used voltage-clamp techniques in Xenopus oocytes and hippocampal neurons to characterize dipicrylamine (DPA) and tetraphenylborate (TPB), compounds previously used to probe membrane structure and voltage-gated ion channel function. KEY RESULTS: Both DPA and TPB potently antagonized GABA(A) receptors. DPA exhibited an IC50 near 60 nM at half-maximal GABA concentration and antagonism with features indistinguishable from pregnenolone sulphate antagonism, including sensitivity to a point mutation in transmembrane domain 2 of the α1 subunit. Bovine serum albumin, which scavenges free membrane-associated DPA, accelerated both capacitance offset and antagonism washout. Membrane interactions and antagonism were explored using the voltage-dependent movement of DPA between membrane leaflets. Washout of DPA antagonism was strongly voltage-dependent, paralleling DPA membrane loss, although steady-state antagonism lacked voltage dependence. At antagonist concentrations, DPA failed to affect inhibitory post-synaptic current (IPSC) amplitude or decay, but DPA accelerated pharmacologically prolonged IPSCs. CONCLUSIONS AND IMPLICATIONS: Neurosteroid-like GABA(A) receptor antagonism appears to lacks a conventional binding site. These features highlight key roles of membrane interactions in antagonism. Because its membrane mobility can be controlled, DPA may be a useful probe of GABA(A) receptors, but its effects on excitability via GABA(A) receptors raise caveats for its use in monitoring neuronal activity.

Structurally diverse amphiphiles exhibit biphasic modulation of GABAA receptors: similarities and differences with neurosteroid actions.

BACKGROUND AND PURPOSE: Some neurosteroids, notably 3alpha-hydroxysteroids, positively modulate GABA(A) receptors, but sulphated steroids negatively modulate these receptors. Recently, other lipophilic amphiphiles have been suggested to positively modulate GABA receptors. We examined whether there was similarity among the actions of these agents and the mechanisms of neurosteroids. Significant similarity would affect theories about the specificity of steroid actions. EXPERIMENTAL APPROACH: Xenopus laevis oocytes were challenged with Triton X-100, octyl-beta-glucoside, capsaicin, docosahexaenoic acid and sodium dodecyl sulphate (SDS), along with different GABA concentrations. KEY RESULTS: These compounds have both positive and negative effects on GABA currents, which can be accentuated according to the degree of receptor activation. A low GABA concentration (1 microM) promoted potentiation and a high concentration (20 microM) promoted inhibition of current, except for SDS that inhibited function even at low GABA concentrations. Amphiphile inhibition was characterized by enhanced apparent desensitization and by weak voltage dependence, similar to pregnenolone sulphate antagonism. We then tested amphiphile effects on mutated receptor subunits that are insensitive to negative (alpha1V256S) and positive (alpha1Q241L or alpha1N407A/Y410F) steroid modulation. Negative regulation by amphiphiles was nearly abolished in alpha1V256S-mutated receptors, but potentiation was unaffected. In alpha1Q241L- or alpha1N407A/Y410F-mutated receptors, potentiation by amphiphiles remained intact. CONCLUSIONS AND IMPLICATIONS: Structurally diverse amphiphiles have antagonist actions at GABA(A) receptors very similar to those of sulphated neurosteroids, while the potentiating mechanisms of these amphiphiles are distinct from those of neurosteroid-positive modulators. Thus, such antagonism at GABA(A) receptors does not have a clear pharmacophore requirement.

Modulation of cerebral vascular tone by activated glia: involvement of nitric oxide.

The ability of activated glia to affect cerebral vascular tone has been evaluated using an in vitro experimental system in which basilar arteries were incubated with glial cultures activated by treatment with lipopolysaccharide (LPS). Vascular tone was measured with an isometric myograph. Contraction in response to high KCl and serotonin was reduced in arteries co-incubated for 24 h with LPS-activated glia, whereas the response to acetylcholine was not modified. The reduced contraction was prevented when the nitric oxide synthase (NOS) inhibitor L-N-nitro-arginine (L-NNA) was added throughout the whole incubation time (activation of glial cells with LPS + co-incubation of glial cells with cerebral arteries). Under these conditions, nitrite levels were drastically reduced. A reduced contraction to KCl was also observed after treatment of the cerebral vessel with sodium nitroprusside. In contrast, L-NNA added to the vessel did not modify the response to contracting stimuli and the expression of endothelial NOS was not modified in cerebral arteries pre-incubated with activated glia. These results suggest that activated glia, which finds an in vivo correlate in several neuropathological conditions, can contribute to changes of vascular tone by modifying the levels of nitric oxide (NO) to which the vessel is exposed.

Novel neuronal targets for the acetylcholinesterase inhibitor donepezil.

The effects of the acetylcholinesterase inhibitor donepezil on cell viability and proliferation events have been analysed in SH-SY5Y human neuroblastoma cells. Short- (48 h) or long-term (7 days) exposure of SH-SY5Y cells to donepezil (100 nM-10 microM) induced a concentration-dependent inhibition of cell proliferation that was not modified by muscarinic and nicotinic receptor antagonists, or mimicked by galantamine, and was not related to induction of apoptosis. By analysing the distribution profile of cell populations within the cell cycle following treatment with 10 microM donepezil, a reduction of cells in the S-G2/M phases of the cycle and a parallel increase of the G0/G1 population were observed. In addition, the expression of two cyclins of the G1/S and G2/M transitions, cyclin E and cyclin B, was significantly reduced in donepezil-treated cells. In contrast, the expression of the cell cycle inhibitor p21 rapidly (6 h) increased following exposure to the drug. Finally, donepezil increased the expression of the neuronal marker MAP-2 in selected subpopulations of SH-SY5Y cells, suggesting that the effect on cell proliferation by donepezil may correlate to a trend to neuronal differentiation.

Glia mediates the neuroprotective action of estradiol on beta-amyloid-induced neuronal death.

17beta-Estradiol (17beta-E(2)) is known to exert neuroprotective activity against beta-amyloid, but its exact target and mechanism of action in this effect have not been elucidated. The involvement of astroglia in neuroprotection of 17beta-E(2) against the beta-amyloid fragment [betaAP((25-35))] has been evaluated using an experimental paradigm in which medium conditioned from rat astroglia pretreated with 17beta-E2 was transferred to pure rat cortical neurons challenged with 25 microm betaAP((25-35)) for 24 h. The toxicity of betaAP((25-35)) was assessed by flow cytometry, evaluating the ability of the peptide to induce an aberrant mitotic cell cycle in neurons. The results obtained indicate that conditioned medium from astrocytes preexposed to 17beta-E(2) for 4 h increased the viability of cortical neurons treated with betaAP((25-35)). This effect was not modified by treatment with the estrogen receptor antagonist ICI 182,780, added directly to neurons, nor was it mimicked by direct addition of 17beta-E(2) to neuronal cultures during exposure to betaAP((25-35)). A soluble factor stimulated by 17beta-E(2) seemed to be involved, and accordingly, the intracellular and released levels of TGF-beta1 were increased by 17beta-E(2) treatment, as established by Western blot analysis. In addition, the intracellular content of TGF-beta1 in immunopositive cells, as detected by flow cytometry, was reduced, suggesting that 17beta-E(2) stimulated mainly the release of the cytokine. In support of a role for TGF-beta1 in astrocyte-mediated 17beta-E(2) neuroprotective activity, incubation with a neutralizing anti-TGF-beta1 antibody significantly modified the reduction of neuronal death induced by 17beta-E(2)-treated astrocyte-conditioned medium.

[Laparoscopic surgery: from clinic to legal medicine]. / Chirurgia Laparoscopica: dalla clinica alla medicina legale.

The laparoscopic technique introduced a new way of operating but inevitably causing new problems for the surgeon. After a comprehensive review of the history and the evolution of laparoscopic surgery from its beginning, the technical aspects of minimally invasive surgery and its fields of application are described. The close dependence on instruments and technology is emphasized. A detailed analysis of the advantages and limitations of laparoscopy is made with emphasis on the importance of a risk-benefit evaluation by the health care provider. Of key importance is to obtain a detailed and clear informed consent. The medico legal aspects of intraoperative complications and the liability of the surgical team in case of patients' injury or death are examined. However, it is always necessary to consider if the potential complications are predictable and/or preventable in accordance to the parameters of negligence, imprudence and lack of knowledge. The same criteria have to be applied to assure compliance with the preventive sanitary rules and that the conversion to laparotomy has been promptly carried out.

Selective thromboxane synthetase inhibition by picotamide and effects on endotoxin-induced lethality.

The efficacy of N,N'-bis-(3-picolyl)-methoxyisophthalamide (picotamide) as an in vitro thromboxane synthetase inhibitor and its effect on endotoxin (LPS)-induced lethality in rats were assessed. Picotamide at 0.5 and 1.0 mM concentrations significantly (P less than 0.05) inhibited basal and LPS-stimulated synthesis of TxA2 measured by its stable immunoreactive (i) metabolite TxB2 in rat peritoneal macrophages. This compound did not inhibit synthesis of i6-keto-PGF1 alpha, the stable metabolite of PGI2, and produced significant shunting to i6-keto-PGF1 alpha. For lethality studies rats were pretreated, by gavage with picotamide, at either 75, 150, 300, or 600 mg/kg 2 hr prior to iv S. enteritidis (LPS, 20 mg/kg). Both 150 and 300 mg/kg doses of picotamide significantly (P less than 0.05) improved survival in endotoxin shock at 48 hr. These studies demonstrate that picotamide is a selective thromboxane synthetase inhibitor, and that it may be useful during disease states characterized by increased TxA2 synthesis.

Effects of different bacterial endotoxins on the drinking behaviour of the rat.

Endotoxins of Pasteurella multocida, Pseudomonas aeruginosa, Neisseria gonorrhoeae, and Shigella sonnei, when given in a single intravenous injection in the rat, showed antidipsogenic effects on drinking behaviour stimulated by 48 h water deprivation, or by intracerebroventricular injection of carbachol (250 ng). In water deprived rats, the antidipsogenic effect was dose related. When drinking was induced by carbachol, endotoxins showed a very long-lasting inhibition. The effect was neither a consequence of behavioural alterations, nor due endotoxin peripheral vasodilatating properties.