Gender differences in microRNA expression in levodopa-naive PD patients.

Gender is an important factor influencing epidemiological and clinical features of Parkinson's disease (PD). We aimed to evaluate gender differences in the expression of a panel of miRNAs (miR-34a-5p, miR-146a, miR-155, miR-29a, miR-106a) possibly involved in the pathophysiology or progression of disease. Serum samples were obtained from 104 PD patients (58 men and 46 women) never treated with levodopa. We measured levels of miRNAs using quantitative PCR. Correlations between miRNA expression and clinical data were assessed using the Spearman's correlation test. We used STRING to evaluate co-expression relationship among target genes. MiR-34a-5p was significantly upregulated in PD male patients compared to PD female patients (fc: 1.62; p < 0.0001). No correlation was found with age, BMI, and disease severity, assessed by UPDRS III scale, in male and female patients. MiR-146a-5p was significantly upregulated in female as compared to male patients (fc: 3.44; p < 0.0001) and a significant correlation was also observed between disease duration and mir-146a-5p. No differences were found in the expression of miR-29a, miR-106a-5p and miR-155 between genders. Predicted target genes for miR-34a-5p and miR-146-5p and protein interactions in biological processes were reported. Our study supports the hypothesis that there are gender-specific differences in serum miRNAs expression in PD patients. Follow-up of this cohort is needed to understand if these differences may affect disease progression and response to treatment.

Evaluation of an optimized [18 F]fluoro-deoxy-glucose positron emission tomography voxel-wise method to early support differential diagnosis in atypical Parkinsonian disorders.

BACKGROUND AND PURPOSE: Atypical Parkinsonian disorders (APD) frequently overlap in clinical presentations, making the differential diagnosis challenging in the early stages. The present study aimed to evaluate the accuracy of the [18 F]fluoro-deoxy-glucose positron emission tomography Statistical Parametric Mapping (SPM) optimized procedure in supporting the early and differential diagnosis of APD. METHODS: Seventy patients with possible APD were retrospectively included from a large clinical cohort. The included patients underwent [18 F]fluoro-deoxy-glucose positron emission tomography within 3 months of the first clinical assessment and a diagnostic follow-up. An optimized SPM voxel-wise procedure was used to produce t-maps of brain hypometabolism in single subjects, which were classified by experts blinded to any clinical information. We compared the accuracy of both the first clinical diagnosis and the SPM t-map classifications with the diagnosis at follow-up as the reference standard. RESULTS: At first diagnosis, 60% of patients were classified as possible APD (progressive supranuclear palsy, corticobasal degeneration, dementia with Lewy bodies, multiple system atrophy) and about 40% as APD with uncertain diagnosis, providing 52% sensitivity, 97% specificity and 86% accuracy with respect to the reference standard. SPM t-map classification showed 98% sensitivity, 99% specificity and 99% accuracy, and a significant agreement with the diagnosis at follow-up (P < 0.001). CONCLUSIONS: The SPM t-map classification at entry predicted the second diagnosis at follow-up. This indicates its significantly superior role for an early identification of APD subtypes, particularly in cases of uncertain diagnosis. The use of a metabolic biomarker at entry in the instrumental work-up of APD may shorten the diagnostic time, producing benefits for treatment options and support to the patients.

Striatal hand in Parkinson's disease: the re-evaluation of an old clinical sign.

Among postural abnormalities in Parkinson's disease (PD), striatal hand (SH) is a particularly underexplored phenomenon. It leads to extreme abnormalities of hand posture, causing altered dexterity, pain and disfigurement. In our study, three blinded investigators examined several pictures of the hands of individuals with PD (N = 40) and controls (N = 15). The investigators quantified postural alterations using the Striatal Hand Score. Demographic and clinical data were also collected. As no differences were detected among investigators agreement, a final Hand Score (HS, range 0-4) was obtained for each hand. The Striatal Hand Score in both the left and right hand was significantly different in PD compared to controls (p < 0.001 for both left and right hand). Striatal hand was significantly worse on the side of PD onset, and on the side with greater PD symptomatology. The finding of a striatal hand was 100 % specific for a diagnosis of PD. Nine PD subjects were evaluated both on and off medication, and dopaminergic treatment did not significantly change the Striatal Hand Score. Our findings suggest that in patients without any explanation for hand deformities other than PD, striatal hand occurs very often, and is highly specific for the side of worst PD involvement. We recommend including an evaluation for SH as part of routine practice. This study emphasizes the importance of a careful observation of the patient in order to improve diagnostic accuracy.

Excitatory deep repetitive transcranial magnetic stimulation with H-coil as add-on treatment of motor symptoms in Parkinson's disease: an open label, pilot study.

BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) has been proposed as a potential treatment for Parkinson's disease (PD). H-coils, inducing deeper and wider magnetic fields compared to traditional coils, may be potentially useful in PD, characterized by widespread, bilateral involvement of cortico-subcortical circuits. OBJECTIVE: To evaluate the safety of repetitive deep TMS (rDTMS) with H-coil as add-on treatment of motor symptoms in PD. METHODS: Twenty-seven PD patients (aged 60.1 ± 6.8 y; PD-duration: 6.3 ± 2.8 y; motor-UPDRS: 39.6 ± 10.1) underwent 12 rDTMS sessions over 4 weeks at excitatory (10 Hz) frequency over primary motor (M1) and bilateral prefrontal (PF) regions. Motor UPDRS off therapy was assessed before and after the last rDTMS session, together with safety records at each treatment session. RESULTS: No drop-outs or adverse events were recorded. Motor UPDRS significantly improved after rDTMS (10.8 points average reduction; P < 0.0001). CONCLUSIONS: High-frequency rDTMS might be a safe treatment for PD motor symptoms. Further placebo-controlled, randomized studies are warranted.

Painful cervical dystonia triggered by the extension wire of a deep brain stimulator.

Deep brain stimulation (DBS) can be complicated by adverse events, which are generally classified as surgical-hardware or stimulation-related. Here we report the onset of a painful cervical dystonia probably triggered by the extension wire of a subthalamic nucleus (STN)-DBS device in a woman suffering from advanced Parkinson's disease (PD). Two months after implantation of the STN-DBS device, our patient developed a painful cervical dystonia, which was not responsive to neurostimulation or to medication. No sign of infections or fibrosis was detected. A patch test with the components of the device was performed, revealing no hypersensibility. The patient was referred back to surgery to reposition the pulse generator in the contralateral subclavian region. A deeper channeling of the wire extensions produced a complete remission of the painful dystonia.

Changes in brain glucose metabolism in subthalamic nucleus deep brain stimulation for advanced Parkinson's disease.

BACKGROUND: Despite its large clinical application, our understanding about the mechanisms of action of deep brain stimulation of the subthalamic nucleus is still limited. Aim of the present study was to explore cortical and subcortical metabolic modulations measured by Positron Emission Tomography associated with improved motor manifestations after deep brain stimulation in Parkinson disease, comparing the ON and OFF conditions. PATIENTS AND METHODS: Investigations were performed in the stimulator off- and on-conditions in 14 parkinsonian patients and results were compared with a group of matched healthy controls. The results were also used to correlate metabolic changes with the clinical effectiveness of the procedure. RESULTS: The comparisons using Statistical parametric mapping revealed a brain metabolic pattern typical of advanced Parkinson disease. The direct comparison in ON vs OFF condition showed mainly an increased metabolism in subthalamic regions, corresponding to the deep brain stimulation site. A positive correlation exists between neurostimulation clinical effectiveness and metabolic differences in ON and OFF state, including the primary sensorimotor, premotor and parietal cortices, anterior cingulate cortex. CONCLUSION: Deep brain stimulation seems to operate modulating the neuronal network rather than merely exciting or inhibiting basal ganglia nuclei. Correlations with Parkinson Disease cardinal features suggest that the improvement of specific motor signs associated with deep brain stimulation might be explained by the functional modulation, not only in the target region, but also in surrounding and remote connecting areas, resulting in clinically beneficial effects.

Neurodegeneration in the course of Langerhans cell histiocytosis.

Cerebral involvement in the course of Langerhans cell histiocytosis has been described especially in children. It is mainly characterized by hypothalamic-pituitary functional deficit, due to granuloma growth. Here we describe a rare case of adult-onset histiocytosis developing a neurodegenerative disease resembling multiple system atrophy. The patient we describe here started suffering from subtle personality changes which progressed to a severe neurological syndrome 2 years after the diagnosis of histiocytosis. Twenty years before she developed a diabetes insipidus, without any apparent cause. Brain MRI scans at the time of neurodegeneration revealed slight signal alterations at the cerebellum, especially involving the dentate nuclei and the white matter. Despite being rare, histiocytosis should be considered in adult patients with cerebellar abnormalities and/or with unexplained diabetes insipidus to rapidly discern and treat histiocytosis before the onset of its neurodegenerative, untreatable phase.

Is saliva suitable as a biological fluid in relative bioavailability studies? Analysis of its performance in a 4 x 2 replicate crossover design.

The aim of the present study was to evaluate the suitability of saliva as a biological fluid in relative bioavailability (RBA) studies, with the focus on the statistical design and data variability. A randomized, open-label, four periods and two sequences (4 × 2) crossover RBA study in saliva of two phenytoin (PHT) 100 mg immediate-release capsules was performed. PHT is a narrow therapeutic index drug that has been widely used for epilepsy treatment for many years. Published information regarding its bioavailability is available, but plasma assessed. This study was designed and performed using saliva as the biological fluid and the simplest conditions that produce coherent results with previously published plasma studies. Pharmacokinetic parameters (C (max), T (max), AUC(0-t ), AUC(0-inf), C (max)/AUC(0-t ), K (e), and t (1/2)) for each volunteer at each period were calculated. Four different BE calculations were performed: individual bioequivalence, by the method of moments, and three average bioequivalence with data averaged over the two administrations and with data of periods 1-2 and 3-4. ANOVA calculation showed no significant subject-by-formulation interaction, period and sequence effects. The intra-subject variabilities were at least 20-fold lower than the inter-subject ones for C (max), AUC(0-t ) and AUC(0-inf). In all four BE calculations, the 90% CIs for the T/R ratios of studied pharmacokinetics parameters fell within the 80-125% range proposed by most regulatory agencies.

The use of saliva as a biological fluid in relative bioavailability studies: comparison and correlation with plasma results.

The aim of the present study was to present new evidence supporting the use of saliva as a biological fluid in relative bioavailability studies. Carbamazepine was chosen as a model drug because of its suitability for salivary therapeutic drug monitoring and its well-documented plasma bioavailability. A relative bioavailability study of four different immediate release carbamazepine products was performed. Stimulated saliva samples were collected by chewing on parafilm wax and by the spitting method. In vitro dissolution testing of formulations, using 900 ml of 1% sodium lauryl sulphate in water, was also carried out. The in vitro-in vivo correlations obtained in this salivary study were consistent with previous correlations assessed using plasma. These results support the suitability of saliva as the biological fluid in relative bioavailability studies.

Therapeutic options and patterns of prescription in chronic venous disorders: results of a 3-year survey in Italy.

OBJECTIVES: To assess self-management of chronic venous disorders (CVDs) in a selected Italian population and the pattern of prescription by selected Italian phlebologists. DESIGN: Cross-sectional study carried out between 2003 and 2005. MATERIALS: Non-random, transverse sample of men and women recruited by advertising. METHODS: Assessment of therapeutic habits of respondents, treatment advice given by phlebologists related to socio-demographic variables and severity of the disease. Multivariate odds ratios for sex, age, class, region, family history and severity of the disease. RESULTS: Women undergo CVD therapy more than men (odds ratio (OR): 2.37 for medical treatment; 1.29 for surgical treatment and 5.72 for sclerotherapy). Young people prefer drug treatment to compression stockings. Drug therapy for CVD is 1.5 times more likely in southern Italian respondents, as is compression stockings (OR: 1.91). Surgical therapy is more frequent in Northern Italy (OR for Central Italy: 0.79; Southern Italy and Islands: 0.76). Family history of CVD leads people to early treatment of symptoms. CONCLUSIONS: This study provides insight into self-medication of CVD in Italy and the prescribing patterns of Italian phlebologists in the treatment of CVD. It shows that the population interviewed is able to practise responsible self-medication of their CVD problems.

Mitochondrial role in ischemia-reperfusion of rat hearts exposed to high-K+ cardioplegia and clonazepam: energetic and contractile consequences.

The role of the mitochondrial Na/Ca-exchanger (mNCX) in hearts exposed to ischemia-reperfusion (I/R) and pretreated with cardioplegia (CPG) was studied from a mechano-calorimetric approach. No-flow ischemia (ISCH) and reperfusion (REP) were developed in isolated rat hearts pretreated with 10 micromol/L clonazepam (CLZP), an inhibitor of the mNCX, and (or) a high K+ - low Ca2+ solution (CPG). Left ventricular end diastolic pressure (LVEDP), pressure development during beats (P), and the steady heat release (Ht) were continuously measured and muscle contents of ATP and PCr were analyzed at the end of REP. During REP, Ht increased more than P, reducing muscle economy (P/Ht) and the ATP content. CPG induced an increase in P recovery during REP (to 90% +/- 10% of preISCH) with respect to nonpretreated hearts (control, C, to 64% +/- 10%, p < 0.05). In contrast, CLZP reduced P recovery of CPG-hearts (50% +/- 6.4%, p < 0.05) and increased LVEDP in C hearts. To evaluate effects on sarcoplasmic reticulum (SR) function, ischemic hearts were reperfused with 10 mmol/L caffeine -36 mmol/L Na (C - caff - low Na). It increased LVEDP, which afterwards slowly relaxed, whereas Ht increased (by about 6.5 mW/g). CLZP sped up the relaxation with higher DeltaHt, C - caff - low Na produced higher contracture and lower Ht in perfused than in ischemic hearts. Values of DeltaHt were compared with reported fluxes of Ca2+-transporters, suggesting that mitochondria may be in part responsible for the DeltaHt during C - caff - low Na REP. Results suggest that ISCH-REP reduced the SR store for the recovery of contractility, but induced Ca2+ movement from the mitochondria to the SR stores. Also, mitochondria and SR are able to remove cytosolic Ca2+ during overloads (as under caffeine), through the mNCX and the uniporter. CPG increases Ca2+ cycling from mitochondria to the SR, which contributes to the higher recovery of P. In contrast, CLZP produces a deleterious effect on ISCH-REP associated with higher heat release and reduced resynthesis of high energy phosphates, which suggests the induction of mitochondrial Ca cycling and uncoupling.

Demographic factors and their relationship with the presence of CVI signs in Italy: the 24-cities cohort study.

BACKGROUND: Chronic venous insufficiency (CVI) is a common progressive disease, the deterioration rate depending on specific risk factors. Demographic characteristics associated with the prevalence of objective signs of CVI were evaluated in a cross-sectional population study in Italy. METHODS: A sample of 5247 people selected by advertising in television and newspapers in 24 Italian cities underwent a clinical examination of the lower limbs, including colour-coded duplex ultrasonography, to determine the presence and severity of CVI. Of these, 4288 subjects provided demographic data that could be analysed by multiple logistic regression model using sex, age, region and family history as covariates. RESULTS: Women were four or more times as likely as men to develop telangiectasia, whilst males had a two-fold increased risk of trunk varices. Age was the main risk factor for vein varicosis in the female population, with women aged over 50 years being almost five times as likely as those aged 29 or less to show trunk varices. Females living in the southern regions had a two-fold increase in risk of developing CVI signs and the risk increased by at least 1.3 times in multiparous women. A positive familial history of disease increased the risk for varicose veins. CONCLUSIONS: Demographic characters such as sex, advanced age, living in the southern regions, number of pregnancies and a family history of CVI were all contributing risk factors for the development of CVI in Italy.

Chronic venous insufficiency in Italy: the 24-cities cohort study.

OBJECTIVE: To assess the relationship between sex, age, geographical region, lower limb symptoms and the presence of trunk varicose veins and venous incompetence. DESIGN: Cross-sectional population study in 24 cities in the North, centre and South of Italy. PARTICIPANTS: Five thousand two hundred and forty-seven people were selected during spring and summer 2003 by advertising on television, in newspapers, and by leaflets in 24 Italian cities. In all 5187 (4457 [85.9%] women and 730 men [14.1%]) volunteers were assessed. The median age was 54 (range: 18-90) years for the women and 61 (range: 18-89) years for the men. METHODS: Self-administered questionnaire on subjective symptoms of chronic venous insufficiency (CVI) in the lower limbs, and clinical examination, including colour duplex ultrasonography to assess the presence and severity of varicose veins. RESULTS: Overall only 22.7% of the subjects examined were free of visible signs of venous disease, with approximately 53% of the population over 50 years of age showing some venous reflux. People living in Southern Italy were more severely affected than those living in the North. Varicosities and telangiectases were the most frequent objective signs in both sexes. Trunk varicosities (27%) and saphenous reflux (41%) increased with age and were more common in men; in contrast, minor objective symptoms such as telangiectases (70%), as well as subjective symptoms such as heavy (79%) and tired legs (78%), were more common in women and were not age-related. CONCLUSIONS: Venous disease is very common in Italy, in particular in people living in the South. A correlation between varicose veins and venous incompetence is more marked in men, while minor objective and subjective symptoms prevail in women. The findings from this non-random sample closely match results from previous studies, in which random sampling was used.

Participation of Na/Ca-exchanger and sarcoplasmic reticulum in the high [K]-protection against ischaemia-reperfusion dysfunction in rat hearts.

AIM: Na/Ca-exchanger (NCX) and sarcoplasmic reticulum (SR) roles during the protection by a cardioplegic solution (25 mm K and 0.5 mm Ca, CPG) against ischaemia-reperfusion was studied. METHODS: Contractile performance (CP) and high energy phosphates contents (HEP) were evaluated in isolated ventricles from rats. They were pre-treated with Krebs (C) or CPG and submitted to no-flow ischaemia and reperfusion (I-R). KB-R7943 5 microm (inhibitor of NCX in reverse mode), 8 mm caffeine and ionic changes were used pre-ischaemically to evaluate each pathway role. RESULTS: During R, CP recovered to 77 +/- 8% of basal in CPG-hearts vs. 55 +/- 8% (P < 0.05) in C-ones. CPG avoided the increases in end diastolic pressure (LVEDP) and in PCr/ATP ratio during I-R. Low [Na]o (78 mm) under both, CPG-2 mm Ca and C, increased further the LVEDP during I-R. LVEDP was also transiently increased by caffeine-CPG, but not modified by KB-R7943. The recovery of CP during reperfusion of CPG-hearts was decreased either, by caffeine (to approximately 75%), low [Na]o-2 mm Ca-CPG (to approximately 40%) and KB-R7943 (to approximately 16%). CONCLUSIONS: CPG protected hearts from ischaemic contracture by attenuating the fall in ATP and removing diastolic Ca by means of NCX in forward mode. Moreover, CPG induces higher CP recovery during reperfusion by participation of SR and NCX in reverse mode. This work remarks the use of CPG based on the functional role of these Ca handling-mechanisms in a pathophysiological condition as ischaemia-reperfusion.

Development of an HPLC method for determination of metabolic compounds in myocardial tissue.

The determination of adenine nucleotides and creatine compounds has great importance in the characterization of ischemic myocardial injury and post-ischemic recovery. It was developed by an HPLC method for the quantification of creatine (Cr), creatine phosphate (CrP), hypoxanthine (HX), AMP, adenosine (Ad), ADP and ATP in isolated perfused rat hearts. The chromatographic conditions were: RP 18 column; mobile phase composed by KH(2)PO(4) (215 mM), tetrabutylammonium hydrogen sulfate (2.3mM), acetonitrile (4%) and KOH (1M 0.4%); flow rate 1 ml min(-1); temperature 25 degrees C; injection volume 20 microl; detection at 220 nm and height peak (HP) as the integration parameter. The method was validated by means of linearity and sensitivity evaluations, using calibration curves done with five concentration levels of each compound. The limits of quantification (LOQ) were also determined. The system precision was calculated as the coefficient of variation for five injections for each compound tested. The purity of the peaks was established using enzymatic peak shift analysis with hexokinase and creatine kinase and also comparing HP at various wavelengths. Frozen hearts were homogenized with a mechanical homogenizer for 3 min at 0 degrees C added with 5 ml of 0.4N HCLO(4). After precipitation with 0.8 ml of 2M KOH the extract was shaked for 2 min and later centrifuged at 0 degrees C for 10 min. The supernatant was kept on ice, filtrated and injected into the HPLC system. The results show that the method for the determination of Cr, CrP, HX, AMP, Ad, ADP and ATP by HPLC here described has good linearity, LOQ, precision, specificity and is simple and rapid to perform.