Candidate biomarkers from the integration of methylation and gene expression in discordant autistic sibling pairs

While the genetics of autism spectrum disorders (ASD) has been intensively studied, resulting in the identification of over 100 putative risk genes, the epigenetics of ASD has received less attention, and results have been inconsistent across studies. We aimed to investigate the contribution of DNA methylation (DNAm) to the risk of ASD and identify candidate biomarkers arising from the interaction of epigenetic mechanisms with genotype, gene expression, and cellular proportions. We performed DNAm differential analysis using whole blood samples from 75 discordant sibling pairs of the Italian Autism Network collection and estimated their cellular composition. We studied the correlation between DNAm and gene expression accounting for the potential effects of different genotypes on DNAm. We showed that the proportion of NK cells was significantly reduced in ASD siblings suggesting an imbalance in their immune system. We identified differentially methylated regions (DMRs) involved in neurogenesis and synaptic organization. Among candidate loci for ASD, we detected a DMR mapping to CLEC11A (neighboring SHANK1) where DNAm and gene expression were significantly and negatively correlated, independently from genotype effects. As reported in previous studies, we confirmed the involvement of immune functions in the pathophysiology of ASD. Notwithstanding the complexity of the disorder, suitable biomarkers such as CLEC11A and its neighbor SHANK1 can be discovered using integrative analyses even with peripheral tissues

Insight in cognitive impairment assessed with the Cognitive Assessment Interview in a large sample of patients with schizophrenia

The Cognitive Assessment Interview (CAI) is an interview-based scale measuring cognitive impairment and its impact on functioning in subjects with schizophrenia (SCZ). The present study aimed at assessing, in a large sample of SCZ (n = 601), the agreement between patients and their informants on CAI ratings, to explore patients' insight in their cognitive deficits and its relationships with clinical and functional indices. Agreement between patient- and informant-based ratings was assessed by the Gwet's agreement coefficient. Predictors of insight in cognitive deficits were explored by stepwise multiple regression analyses. Patients reported lower severity of cognitive impairment vs. informants. A substantial to almost perfect agreement was observed between patients' and informants' ratings. Lower insight in cognitive deficits was associated to greater severity of neurocognitive impairment and positive symptoms, lower severity of depressive symptoms, and older age. Worse real-life functioning was associated to lower insight in cognitive deficit, worse neurocognitive performance, and worse functional capacity. Our findings indicate that the CAI is a valid co-primary measure with the interview to patients providing a reliable assessment of their cognitive deficits. In the absence of informants with good knowledge of the subject, the interview to the patient may represent a valid alternative

The interplay among psychopathology, personal resources, context-related factors and real-life functioning in schizophrenia: stability in relationships after 4 years and differences in network structure between recovered and non-recovered patients

Improving real-life functioning is the main goal of the most advanced integrated treatment programs in people with schizophrenia. The Italian Network for Research on Psychoses previously explored, by using network analysis, the interplay among illness-related variables, personal resources, context-related factors and real-life functioning in a large sample of patients with schizophrenia. The same research network has now completed a 4-year follow-up of the original sample. In the present study, we used network analysis to test whether the pattern of relationships among all variables investigated at baseline was similar at follow-up. In addition, we compared the network structure of patients who were classified as recovered at follow-up versus those who did not recover. Six hundred eighteen subjects recruited at baseline could be assessed in the follow-up study. The network structure did not change significantly from baseline to follow-up, and the overall strength of the connections among variables increased slightly, but not significantly. Functional capacity and everyday life skills had a high betweenness and closeness in the network at follow-up, as they had at baseline, while psychopathological variables remained more peripheral. The network structure and connectivity of non-recovered patients were similar to those observed in the whole sample, but very different from those in recovered subjects, in which we found few connections only. These data strongly suggest that tightly coupled symptoms/dysfunctions tend to maintain each other's activation, contributing to poor outcome in schizophrenia. Early and integrated treatment plans, targeting variables with high centrality, might prevent the emergence of self-reinforcing networks of symptoms and dysfunctions in people with schizophrenia

The interplay among psychopathology, personal resources, context-related factors and real-life functioning in schizophrenia: stability in relationships after 4 years and differences in network structure between recovered and non-recovered patients

Improving real-life functioning is the main goal of the most advanced integrated treatment programs in people with schizophrenia. The Italian Network for Research on Psychoses previously explored, by using network analysis, the interplay among illness-related variables, personal resources, context-related factors and real-life functioning in a large sample of patients with schizophrenia. The same research network has now completed a 4-year follow-up of the original sample. In the present study, we used network analysis to test whether the pattern of relationships among all variables investigated at baseline was similar at follow-up. In addition, we compared the network structure of patients who were classified as recovered at follow-up versus those who did not recover. Six hundred eighteen subjects recruited at baseline could be assessed in the follow-up study. The network structure did not change significantly from baseline to follow-up, and the overall strength of the connections among variables increased slightly, but not significantly. Functional capacity and everyday life skills had a high betweenness and closeness in the network at follow-up, as they had at baseline, while psychopathological variables remained more peripheral. The network structure and connectivity of non-recovered patients were similar to those observed in the whole sample, but very different from those in recovered subjects, in which we found few connections only. These data strongly suggest that tightly coupled symptoms/dysfunctions tend to maintain each other's activation, contributing to poor outcome in schizophrenia. Early and integrated treatment plans, targeting variables with high centrality, might prevent the emergence of self-reinforcing networks of symptoms and dysfunctions in people with schizophrenia

The association between insight and depressive symptoms in schizophrenia: Undirected and Bayesian network analyses

Background. Greater levels of insight may be linked with depressive symptoms among patients with schizophrenia, however, it would be useful to characterize this association at symptom-level, in order to inform research on interventions. Methods. Data on depressive symptoms (Calgary Depression Scale for Schizophrenia) and insight (G12 item from the Positive and Negative Syndrome Scale) were obtained from 921 community-dwelling, clinically-stable individuals with a DSM-IV diagnosis of schizophrenia, recruited in a nationwide multicenter study. Network analysis was used to explore the most relevant connections between insight and depressive symptoms, including potential confounders in the model (neurocognitive and social-cognitive functioning, positive, negative and disorganization symptoms, extrapyramidal symptoms, hostility, internalized stigma, and perceived discrimination). Bayesian network analysis was used to estimate a directed acyclic graph (DAG) while investigating the most likely direction of the putative causal association between insight and depression. Results. After adjusting for confounders, better levels of insight were associated with greater self-depreciation, pathological guilt, morning depression and suicidal ideation. No difference in global network structure was detected for socioeconomic status, service engagement or illness severity. The DAG confirmed the presence of an association between greater insight and self-depreciation, suggesting the more probable causal direction was from insight to depressive symptoms. Conclusions. In schizophrenia, better levels of insight may cause self-depreciation and, possibly, other depressive symptoms. Person-centered and narrative psychotherapeutic approaches may be particularly fit to improve patient insight without dampening self-esteem

Rationale and design of an independent randomised controlled trial evaluating the effectiveness of aripiprazole or haloperidol in combination with clozapine for treatment-resistant schizophrenia

<p>Abstract</p> <p>Background</p> <p>One third to two thirds of people with schizophrenia have persistent psychotic symptoms despite clozapine treatment. Under real-world circumstances, the need to provide effective therapeutic interventions to patients who do not have an optimal response to clozapine has been cited as the most common reason for simultaneously prescribing a second antipsychotic drug in combination treatment strategies. In a clinical area where the pressing need of providing therapeutic answers has progressively increased the occurrence of antipsychotic polypharmacy, despite the lack of robust evidence of its efficacy, we sought to implement a pre-planned protocol where two alternative therapeutic answers are systematically provided and evaluated within the context of a pragmatic, multicentre, independent randomised study.</p> <p>Methods/Design</p> <p>The principal clinical question to be answered by the present project is the relative efficacy and tolerability of combination treatment with clozapine plus aripiprazole compared with combination treatment with clozapine plus haloperidol in patients with an incomplete response to treatment with clozapine over an appropriate period of time. This project is a prospective, multicentre, randomized, parallel-group, superiority trial that follow patients over a period of 12 months. Withdrawal from allocated treatment within 3 months is the primary outcome.</p> <p>Discussion</p> <p>The implementation of the protocol presented here shows that it is possible to create a network of community psychiatric services that accept the idea of using their everyday clinical practice to produce randomised knowledge. The employed pragmatic attitude allowed to randomly allocate more than 100 individuals, which means that this study is the largest antipsychotic combination trial conducted so far in Western countries. We expect that the current project, by generating evidence on whether it is clinically useful to combine clozapine with aripiprazole rather than with haloperidol, provides physicians with a solid evidence base to be directly applied in the routine care of patients with schizophrenia.</p> <p>Trial Registration</p> <p><b>Clincaltrials.gov Identifier</b>: NCT00395915</p

MULTIDIMENSIONAL ANALYSIS OF SELF-ESTEEM IN A GROUP OF CHILDREN WITH ADHD

Attention Deficit Hyperactivity Disorder (ADHD) is characterised by low levels of internal-izing symptoms and self-efficacy which causes low self-esteem, while externalizing be-haviours appear to be related to high levels of stress in the parents. The purpose of this research is to analyze the impact of ADHD on self-esteem and paren-tal stress. A multidimensional assessment of self-esteem was performed using the MSCS (Multidimensional Self-Concept Scale) in a group of 12 male patients with ADHD (age range 9-11 years). Parental stress was investigated using the Parenting Stress Index (PSI). These results were compared with a group of 12 healthy children (age 9-11 years), with both parametric statistics and correlation statistics. The comparison between ADHD children and control subjects, performed by a calculation to rank with the Mann-Whitney, showed a high significance in two dimensional compo-nents of self-esteem: social relationships (Z -2.028 p 0.045) and academic success (Z -2.166 p 0.028). The total self-esteem score differed significantly between the two groups (Z -2.227 p 0.024). Parental stress increaseed with the level of the child‟s oppositional symptoms (p 0.790) but it did not correlate with the other scores (cognitive problems / inattention p 0.381); hyperactivity p 0.414; ADHD index p 0.324). The present study shows that self-esteem is impaired among children with ADHD

Nanoassembly of an amphiphilic cyclodextrin and Zn(II)-phthalocyanine with the potential for photodynamic therapy of cancer

Due to their poor solubility and propensity to aggregate in aqueous media, therapeutic application of several photosensiting agents, such as phthalocyanines, in photodynamic therapy (PDT) of solid tumors is severely hampered. With the aim to propose a novel nanotechnological approach, in this paper biodegradable nanoassemblies based on heptakis (2-oligo(ethyleneoxide)-6-hexadecylthio-)-??-CD (SC16OH) and zinc-phthalocyanine (ZnPc) were developed and tested. Nanoassemblies, prepared by the emulsion???solvent evaporation technique, displayed a hydrodynamic diameter around 200 nm, a negative zeta potential and a satisfactory entrapment efficiency of ZnPc. Steady-state and time resolved fluorescence emission spectroscopy studies showed the entrapment of ZnPc as a monomer in the carrier, with a low tendency to self-aggregate and consequently a fairly good propensity to generate singlet oxygen after photoactivation. The interaction of ZnPc with SC16OH was elucidated by 1H-NMR, which suggested the formation of complexes between drug and both hydrophobic and hydrophilic moieties of the amphiphile. Finally, in vitro potential of the nanoassembly was evaluated in HeLa cells by following cellular uptake and photobiological activity. Overall, results suggest the suitability of the nanoassembly based on SC16OH for delivering ZnPc to cancer cells, thus inducing photodynamic anticancer effects

Nanoassemblies based on non-ionic amphiphilic cyclodextrin hosting Zn(II)-phthalocyanine and docetaxel: Design, physicochemical properties and intracellular effects

The combination of conventional anticancer therapy with other treatment modalities such as photodynamic therapy (PDT) is paving the way to novel more effective treatment of solid tumors via light exposure. With this idea in mind, in this paper, nanoparticles (NPs) based on Heptakis (2-oligo(ethyleneoxide)-6-hexadecylthio-)-β-CD (SC16OH) for dual delivery of Zinc-Phthalocyanine (ZnPc) and Docetaxel (DTX) were developed pointing to their potential application as nanomedicine for the combined photodynamic and chemo-therapy of solid tumors. NPs prepared by the emulsion-solvent evaporation technique displayed a hydrodynamic diameter of ≅ 200nm, a negative zeta potential (≅ -27mV) and a satisfactory entrapment efficiency of both drugs at a specific mass ratio. On these bases, NPs containing DTX and ZnPc with theoretical loading of 5% and 0.2% respectively (ZnPc/DTX5-NPs) were selected for further investigations. The allocation of ZnPc and DTX into the colloid was investigated by complementary spectroscopic techniques. In particular, fluorescence emission studies showed the entrapment of ZnPc as a monomer in the carrier, with a low tendency to self-aggregate and consequently a fairly high propensity to photogenerate singlet oxygen. The interaction of SC16OH with DTX, co-entrapped with ZnPc, was elucidated by (1)H NMR and 2D ROESY, which suggested the presence of the chemotherapeutic in the hydrophobic portion of SC16OH. ZnPc/DTX5-NPs were fairly stable in different biological relevant media within 24h. Finally, in vitro potential of the nanoassembly was evaluated in HeLa cancer cells by cell viability exploring both effects of DTX and ZnPc. Overall, results suggest the suitability of NPs based on SC16OH for delivering a combination of DTX with ZnPc to cancer cells, thus inducing photodynamic and antimitotic effects

EFFECTIVENESS OF SACRAL NEUROMODULATION IN THE MANAGEMENT OF REFRACTORY BLADDER PAIN SYNDROME

INTRODUCTION AND AIM OF THE STUDY Bladder Pain Syndrome (BPS) is a complex, under-diagnosed invalidating pathology. It is difficult to estimate BPS prevalence, because this condition is underestimated. A survey estimated the prevalence of BPS symptoms among adult females in the USA to be 7% [1], whereas Chronic Pelvic Pain symptoms show an incidence overlapping to migraine, asthma and lumbago [2]. Treatment of BPS symptoms is an actual therapeutic challenge and needs a multi-disciplinary management. Paracetamol, NSAID's, antidepressant and anti-convulsivants are the first line treatment; in refractory patients, capsaicin, Transcutaneous Electrical Nerve Stimulation (TENS) and sacral neuromodulation (SNM) may be further options. SNM can reduce BPS symptoms and improves voids and over 90% of patients treated with neuromodulation would undergo the implant again [3]. The aim of this study was to evaluate the effectiveness of SNM for the treatment of symptoms in patients with refractory BPS. MATERIALS AND METHODS A retrospective chart review was performed of patients with BPS, refractory to medical/conservative therapy, who underwent InterStim\uae unilateral sacral neuromodulation implant between March 2012 and July 2014 in a single Italian centre. All patients were affected by BPS according to the diagnostic criteria (pelvic pain>6 months, with pressure or discomfort to the bladder, associated to at least one of lower urinary tract symptoms (LUTS). All patients were non responders to oral or endovesical therapy; all patients were>18 year-old and a complete work-up was conducted, in order to exclude organic causes of BPS. Patients with pelvic organ prolapse, stress urinary incontinence or neurologic disease were excluded from the treatment. Bladder pain was assessed with Visual Analogue Scale (VAS) before the treatment, one year after the treatment and at the last follow-up visit. LUTS were evaluated using a three-day bladder diary before the treatment, one year after and in the last follow-up visit. All complications were recorded. RESULTS Twenty-three patients (18 women, and 5 men) underwent first stage unilateral S3 stimulation; median age was 56 years (range 43\u201375 years) and mean follow-up was 32 months. Average symptoms duration was 2.5 years (range 1\u20136.5); 78% of the patients (only women) showed a significant improvement ( 6550% relief of pain and LUTS) and underwent a definitive implant, In all patients, the implantable pulse generator (IPG) was placed in a subcutaneous pouch in the gluteal region, on the right side in 85% of cases. Baseline and post-treatment VAS and parameters of the bladder diary were compared, showing significant improvements. Patients' reported an average pain score decreased from a mean of 8 (9\u201310) at baseline to 3 (2\u20135) at one-year follow-up (p\u2009<\u20090.001). Urgency decreased from 4.6\u2009\ub1\u20092.4 up to 1.3\u2009\ub1\u20091.9 (p\u2009<\u20090.001). Daily urinary frequency improved from 12.7\u2009\ub1\u20094.8 times up to 8.7\u2009\ub1\u20092.8 (p\u2009<\u20090.001) and nocturia decreased from 2.5\u2009\ub1\u20091.9 up to 0.7\u2009\ub1\u20091 (p\u2009<\u20090.001). Mean voided volume increased from 145.4\u2009\ub1\u200970.5\u2009ml to 208.2\u2009\ub1\u200973.4\u2009ml (p\u2009<\u20090.001). Clinical effectiveness was maintained in the last follow-up visit. There were no cases of: infection, pain at the site of implantation, replacement of IPG due to ended battery life. INTERPRETATION OF RESULTS Although literature studies are at high risk of bias and are difficult to compare, due to the different NMS techniques, evaluation of the outcomes and patient selection, our results are at least overlapping with the literature data. CONCLUSIONS Sacral neuromodulation appears to be effective and safe in treating refractory BPS with LUTS associated. Our study provides further evidence for the role of this therapy in BPS, an invalidating pathology with a major impact on patients' quality of life and that is difficult to diagnose and to treat