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Energy efficiency in buildings is a key issue in the current energy transition. In order to reduce building energy consumption, users’ behaviour and the perception of indoor environmental comfort must be taken into account; these aspects are inextricably linked to energy demand, consumption and related costs. In this paper, we present the methodological framework, technological solutions and outcomes of the ComfortSense project. ComfortSense aimed at decoupling energy demand from indoor comfort. We focused on Heating, Ventilating and Air Conditioning (HVAC) systems in buildings, on users’ behaviour and on comfort perception by treating buildings as socio-technical systems. Our approach - which was multidisciplinary and included the contribution of sociologists, physicists and computer scientists - was based on Internet of Things technologies, on a Living Lab design and testing process and on a Crowdsensing approach. Physical parameters (objective variables), such as temperature, CO2 concentration and relative humidity, were measured by a Wireless Sensor Network and by wearable devices, while the users’ perception of comfort (subjective variables) were recorded as real-time feedback through a Mobile App in three pilot buildings of the University of Turin, engaging about a thousand buildings’ users (professors, researchers, students and employees). Objective and subjective variables were correlated through an ad-hoc Direct Virtual Sensor. Thanks to the Direct Virtual Sensor forecasting we demonstrated that, adopting an adaptive indoor comfort management, users’ comfort can be remarkably improved while reducing the energy consumption of HVAC systems.

This contribution is the result of our progressive engagement to develop and to apply a top-down liquid chromatography (LC) matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) (LC-MALDI-TOF) analysis for the histone post-translational modifications (PTMs) and variants characterization, mainly in order to provide comprehensive and fast results. The histone post-translational modifications and the differential expression of the histone variants play an essential role both in the DNA packaging mechanism in chromosomes and in the regulation of gene expression in different cellular processes, also in response to molecular agents of environmental origin. This epigenetic mechanism is widely studied in different field such as cellular differentiation, development and in the understanding of mechanisms underlying diseases. The characterization of histone PTMs has traditionally performed by antibodies-based assay, but immunological methods have significant limits, and today systems that use mass spectrometry are increasingly employed. We evaluated an in-source decay (ISD) analysis for the histone investigation on human lymphoblastoid cells, and by this approach, we were able to identify and quantify several PTMs such as the di-methylation in the lysine 20 and the acetylation in the lysine 16 in H4 and the mono-methylation, di-methylation and trimethylations at K9 of the histone H3.1. Moreover, we detected and quantified in the same H2B spectrum the prevalent H2B 1C/2E type but also the minor H2B 1D, 1M and 1B/1L/1N, 1O/2F, 1J/1K variants. In this work, we show that MALDI-ISD represents an excellent methodology to obtain global information on histone PTMs and variants from cells in culture, with rapidity and simplicity of execution. Finally, this is a useful approach to get label-free relative quantitative data of histone variants and PTMs.

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Background: Neurological disorders are a highly heterogeneous group of pathological conditions that affect both the peripheral and the central nervous system. These pathologies are characterized by a complex and multifactorial etiology involving numerous environmental agents and genetic susceptibility factors. For this reason, the investigation of their pathogenetic basis by means of traditional methodological approaches is rather arduous. High-throughput genotyping technologies, including the microarray-based comparative genomic hybridization (aCGH), are currently replacing classical detection methods, providing powerful molecular tools to identify genomic unbalanced structural rearrangements and explore their role in the pathogenesis of many complex human diseases. Methods: In this report, we comprehensively describe the design method, the procedures, validation, and implementation of an exon-centric customized aCGH (NeuroArray 1.0), tailored to detect both single and multi-exon deletions or duplications in a large set of multi- and monogenic neurological diseases. This focused platform enables a targeted measurement of structural imbalances across the human genome, targeting the clinically relevant genes at exon-level resolution. Conclusion: An increasing use of the NeuroArray platform may offer new insights in investigating potential overlapping gene signatures among neurological conditions and defining genotype-phenotype relationships.

La Sclerosi Multipla è una malattia neurodegenerativa progressiva e infiammatoria, spesso disabilitante, caratterizzata da lesioni demielinizzanti e atrofia del sistema nervoso centrale. La maggior parte delle persone viene diagnosticata tra i 20 ed i 40 anni. La Sclerosi Multipla presenta un decorso variabile con disturbi sensoriali, visivi, alterazioni dell'equilibrio, affaticamento, spasticità, intorpidimento, linguaggio legato, disfunzioni sessuali, impedimento nella deambulazione, disfunzione cognitiva, depressione (Busillo B., 2014; Bronner G., 2010). La scoperta di essere affette da SM apre uno scenario di crisi esistenziale: la persona è costretta a separarsi dal suo stato di vita precedente per affrontare qualcosa di nuovo e sconosciuto, imprevedibile e bizzarro. Scopo del presente studio è stato quello di stimolare le singole persone a recuperare le proprie risorse e metterle a servizio di un progetto personale o di gruppo, come forma di terapia intesa a fronteggiare il senso di impotenza e inutilità che accompagna la SM.

Mutations in the Angiogenin gene (ANG) linked to 14q11.2 have been recently discovered to be associated with Amyotrophic Lateral Sclerosis (ALS) in Irish and Scottish populations. In our study we investigated the role of ANG gene in ALS patients from southern Italy. We found a novel mutation in the signal peptide of the ANG gene in a sporadic patient with ALS (SALS). The molecular analysis of the ANG gene also demonstrated an allelic association with the rs11701 single nucleotide polymorphism (SNP) in familial ALS (FALS) but not in SALS patients. Our finding supports the evidence that the ANG gene is involved in ALS.

Fused in sarcoma (FUS) or translocation in liposarcoma (TLS), a DNA/RNA-binding protein, causes a dominant autosomal inherited form of amyotrophic lateral sclerosis (ALS), ALS 6. Its main role in neurodegeneration is highlighted by the presence of cytoplasmic accumulation of its mutant protein form in ALS patients. To further define the frequency and spectrum of FUS gene mutations, we have performed a molecular screening of a cohort of 327 Italian patients from Southern Italy with sporadic ALS (SALS). We identified 4 patients carrying 3 different missense mutations and several polymorphisms. Two different substitutions occurring in the same amino acidic position have been observed in 2 patients: R521G and R521C respectively; P525L mutation has been found in 2 additional cases. Most of the patients with FUS mutations showed early symptom onset and had short disease survival. We also detected 4 different polymorphic variants (3=-untranslated region [UTR] variant, c.*41G.A; c.52313ins[GAGGTG]; c.335-15del[TTTT]; and rs13331793) in 9 patients from within our cohort. This study underlines the importance of population-based mutation screening of newly identified genes. © 2011 Elsevier Inc. All rights reserved.

Objective: Recent evidence suggests that intermediate-length polyglutamine (PolyQ) expansions in the ataxin-2 ( ATXN-2 ) gene are a risk factor for amyotrophic lateral sclerosis (ALS). This work was undertaken with the aim to investigate the frequency of ataxin-1 ( ATXN-1 ) and ATXN-2 PolyQ expansions in a cohort of patients with sporadic ALS (sALS) and patients with familial ALS (fALS) from southern Italy. Methods: We assessed the PolyQ lengths of ATXN-1 and ATXN-2 in 405 patients with sALS, 13 patients with fALS, and 296 unrelated controls without history of neurodegenerative disorders. Results: We found significantly higher intermediate PolyQ expansions ≥32 for ATXN-1 alleles and ≥28 for ATXN-2 alleles in the sALS cohort ( ATXN-1 : ALS, 7.07% vs controls, 2.38%; p = 0.0001; ATXN-2 : ALS, 2.72% vs controls, 0.5%; p = 0.001). ATXN-1 CAT and ATXN-2 CAA interruptions were detected in patients with ALS only. Age at onset, site of onset, and sex were not significantly related to the ATXN-1 or ATXN-2 PolyQ repeat length expansions. Conclusions: Both ATXN-1 and ATXN-2 PolyQ intermediate expansions are independently associated with an increased risk for ALS.

Introduction.The post-translational (PTMs) modifications of nuclear histones, as well known, play an essential role both in the DNA packaging mechanism in chromosomes that in the regulation of gene expression in different cellular processes, especially in response to molecular agents of environmental origin (epigenetic regulation). Aim.The characterization of histone PTMs is traditionally performed by the use of antibodies. However, these methods are often not sufficiently specific and today systems that use mass spectrometry are increasingly used. In this context, our research aims to improve the methodologies based on mass spectrometry mainly in relation to the comprehensivity, simplicity and rapidity of the characterization. Methods.For this purpose, a top-down analysis of MALDI- In Source Decay (MALDI-ISD) analysis, on HPLC fraction of nuclear extracts obtained from immortalized EBV lymphocytic lines, was applied. The cells grown in RPMI 1640 medium were lysed and the collected nuclear fraction was subjected to acid extraction with 0.8M HCl. A preliminary MALDI-TOF profiling analysis was performed on the whole crude extract, and successively this was subjected to HPLC separation from which homogenous fractions of histones (H2A, H2B, H3.1, H3.2, H3.3 and H4) were obtained. Fractions of interest were analyzed using MALDI-ISD using 1.4 DAN as matrix, in order to obtain sequencing and PTMs data. In some cases, a PSD analysis was also performed on fragments generated by the ISD (MS3) to confirm data. Results.The MALDI-TOF profiling of the crude nuclear extract revealed the presence of the principal types of histones, allowing also a relative quantitative evaluation. Subsequently, the MALDI-ISD analysis of the HPLC fractions provided information on both the amino acid sequence and the PTMs present. In particular, we were able to define, in the same experiment the presence in H4 of a dimethylation (above 90%) in the lysine at position 20 (H4K20Me2) Conclusions.In conclusion, the MALDI-ISD methodology allows to obtain global information on histone PTMs with specificity, rapidity and simplicity, effectively contributing to reveal the influence of environmental molecular factors

Autosomal recessive spastic paraplegia form 35 (SPG35) is a rare form of Hereditary Spastic Paraplegia (HSP, MIM 18260) characterized by childhood onset of spasticity, cognitive decline and leukodystrophy. Additional clinical features such as seizures, dysphagia, dysarthria, dystonia, neuropathy and brain iron accumulation were also observed. Mutations in the fatty acid 2-hydroxylase (FA2H) gene have been associated to the SPG35 form. FA2H encodes a nicotinamide adenine dinucleotide phosphate (NADPH)-dependent monooxygenase, involved in the synthesis of 2-hydroxy fatty acid galactolipids, that are the major component of myelin sheath . In the current study we report two siblings with a complicated form of hereditary spastic paraparesis carrying two deleterious FA2H compound heterozygous missense mutations identified by exome sequencing.

The routine molecular test for spinal muscular atrophy (SMA) diagnosis is based on the detection of a homozygous deletion of exons 7 and 8 of the telomeric copy of the survival motor neuron gene (SMN1). The presence of the centromeric copy of the SMN gene (SMN2) does not allow the detection of the hemizygous absence of the SMN1 gene, which characterizes the disease carriers. The demand for a quantitative SMN1 test is permanently growing because there is a high incidence of carriers. The disease is severe and to date there are no effective pharmacological treatments. Here, we present a non-radioactive assay based on real time quantitative polymerase chain reaction. We analyzed eight SMA patients, 14 SMA relatives and 50 health individuals from Southern Italy by real time quantitative method in order to identify haploid deletion occurring in SMA carriers. SMN1 copy number was determined by the comparative threshold cycle method (ΔΔCt). The results confirmed the deletion in all homozygous patients and permitted an evaluation of the number of alleles in the healthy carriers. This method is fast, reproducible, and enables us to discriminate carriers from healthy homozygous, which is impossible with normal techniques.

Mutations in the Cu/Zn superoxide dismutase 1 (SOD1) gene have been reported to cause adult-onset autosomal dominant amyotrophic lateral sclerosis (FALS). In sporadic cases (SALS), de novo mutations in the SOD1 gene have occasionally been observed. All the SOD1 mutations are autosomal dominantly inherited with the exception of D90A. To date, in Italy, only two sporadic ALS cases carrying the D90A mutation have been reported in a homozygous state. We investigated for the presence of this mutation in 169 unrelated ALS patients from southern Italy. The genetic analysis revealed three ALS patients (1.8%) with mild phenotype carrying the homozygous D90A mutation.

Mutations in the Cu/Zn superoxide dismutase (Sod1) gene have been reported to cause adult-onset autosomal dominant Amyotrophic Lateral Sclerosis (FALS). In sporadic cases (SALS) de novo mutations in the Sod1 gene have occasionally been observed. The recent finding of a mutation in the VAMP/synaptobrevin-associated membrane protein B (VAPB) gene as the cause of amyotrophic lateral sclerosis (ALS8), prompted us to investigate the entire coding region of this gene in SALS patients. One hundred twenty-five unrelated patients with adult-onset ALS and 150 healthy sex-age-matched subjects with the same genetic background were analyzed. Genetic analysis for all exons of the VAPB gene by DHPLC revealed 5 variant profiles in 83 out of 125 SALS patients. Direct sequencing of these PCR products revealed 3 nucleotide substitutions. Two of these were found within intron 3 of the gene, harbouring 4 variant DHPLC profiles. The third nucleotide variation (Asp130Glu) was the only substitution present in the coding region of the VAPB gene, and it occurred within exon 4. It was found in three patients out of 125. The frequency of the detected exon variation in the VAPB gene was not significantly different between patients and controls. In conclusion, our study suggests that VAPB mutations are not a common cause of adult-onset SALS.

Genetic variants within the serotonin transporter gene (5-HTTLPR) impact the neurobiology and risk for anxiety-related behaviours. There are also gender differences in the prevalence of anxiety-related behaviours. Although numerous studies have investigated the influence of 5-HTTLPR genotype on the neural systems involved in emotional regulation, none have investigated how these effects are modulated by gender and anxiety. We investigated this issue using two complementary region of interest-based structural neuroimaging approaches (voxel-based morphometry and Freesurfer) in 138 healthy individuals categorized into 'no anxiety' and 'subclinical anxiety' groups based on the Hamilton Rating Scale for Anxiety (HAM-A). Preliminarily, using anxiety as a continuous variable, we found a significant interaction effect of genotype by gender on anxiety. Females homozygous for the Short allele showed the highest HAM-A scores and males the lowest. In addition, a three-way significant interaction among genotype, gender and anxiety category was found for the right amygdala volume. Post hoc tests revealed that homozygous females carrying the Short variant with a subclinical anxiety condition had larger volume. The reported interaction effects demonstrate that gender strongly modulates the relationship between 5-HTTLPR genotype and subclinical expression of anxiety acting on amygdala, one region of the emotional neural network specifically involved in the anxiety-like behaviours.

La Neurofibromatosi di tipo I (NF1) rappresenta la forma più comune di sindrome neurocutanea. Si tratta di un disordine autosomico dominante, causato da mutazioni in eterozigosi a carico dell'oncosoppressore NF1 sul cromosoma 17q11.2,costituito da 57 esoni costitutivi e 3 che vanno incontro a splicing alternativo. Tra le mutazioni patogenetiche riscontrate vi sono delezioni parziali o dell'intero gene. NF1, pur essendo un disordine mendeliano a penetranza completa, si caratterizza per un'alta variabilità fenotipica; infatti sono stati descritti fenotipi clinici più severi in pazienti con delezioni che coinvolgono interamente il gene NF1 e quelli fiancheggianti, rispetto a pazienti con mutazioni intrageniche. In questo lavoro, presentiamo lo sviluppo di un array custom, esone-centrico e ad alta risoluzione, per ibridazione genomica comparativa (aCGH). NeuroArray 1.0. è stato progettato per l'identificazione di varianti del numero di copie (CNV) in 1.632 geni di interesse in differenti disordini neurologici, tra i quali alcune sindromi neurocutanee come le Neurofibromatosi di tipo 1 e 2, e le Sclerosi tuberose di tipo 1 e 2. Tale array è stato sviluppato su piattaforma Agilent Technologies 8x60K. In particolare, descriviamo la validazione della regione del cromosoma 17 che circonda e include NF1, coprendo un intervallo di ~2.7 Mb con una distanza mediana tra le sonde di 173.3 bp. Tale regione è stata validata utilizzando 5 campioni di DNA di pazienti con diagnosi clinica di NF1, risultati negativi ad uno screening mutazionale del gene. Con il NeuroArray 1.0 sono state identificate due delezioni, una intragenica ed una atipica che si estende a valle del gene. Tali delezioni erano state precedentemente identificate mediante MLPA in due dei cinque pazienti esaminati. Paragonando il NeuroArray 1.0 con altre piattaforme aCGH custom per NF1, abbiamo ottenuto una singola piattaforma in grado di analizzare a) un vasto numero di geni correlati a disordini neurologici, b) il gene NF1 ed i geni fiacheggianti con risoluzione esonica, c) 8 campioni per volta. La piattaforma NeuroArray 1.0 può essere dunque utilizzata sia per la ricerca di mutazioni strutturali sbilanciate in pazienti affetti da NF1 che come test di conferma di dati ottenuti mediante NGS.

Hereditary spastic paraplegia (HSP, MIM 18260) comprises a group of inherited neurodegenerative disorders that are characterized by progressive spasticity and weakness of lower extremities, due to a length dependent dying back degeneration of corticospinal tracts. Clinically, HSP can be classified as pure if spastic paraparesis occurs in isolation, and complex if other neurological or non-neurological features are present. Autosomal dominant (AD), autosomal recessive (AR) and X linked modes of inheritance have been described).Among these, the autosomal recessive spastic paraplegia form 35 (SPG35) is characterized by childhood onset of spasticity, cognitive decline and leukodystrophy). Additional clinical features such as seizures, dysphagia, dysarthria, dystonia, neuropathy and brain iron accumulation were also observed (2-3). Several patients were described with different combination of these features. Mutations in the fatty acid 2-hydroxylase (FA2H) gene have been associated to the SPG35 form. FA2H encodes a nicotinamide adenine dinucleotide phosphate (NADPH)-dependent monooxygenase, involved in the synthesis of 2-hydroxy fatty acid galactolipids, that are the major component of myelin sheath).

Hereditary spastic paraplegias are neurodegenerative disorders characterized clinically by progressive spasticity of the lower limbs; they are inherited as autosomal dominant, autosomal recessive and X-linked traits. We have analyzed four autosomal recessive HSP families gathered from southern Italy. We performed genetic analysis using microsatellite markers associated with SPG5, SPG7, SPG11 and SPG14. Positive lod scores were obtained with markers located on chromosome 8. The lod scores for the four combined families were significantly higher than 3 for D8S509, D8S1102, D8S1723 and D8S260 with a maximum two-point lod score at θ = 0 of 3.99 for the marker D8S260. In one of the examined families, the haplotype analysis suggests two key recombination events demonstrating that the gene is localized in the 11 cM region flanked by markers D8S285 and D8S544, refining the ARHSP region by approximately 22 cm. We also analyzed five candidate genes localized within the HSP region: TOX, syndecan-binding-protein (SDCBP), RAB2, CA8 and PENK, but we did not find disease causing mutations.

Background Previous evidence has shown that genetic variations in the serotonergic system contribute to individual differences in personality traits germane to impulse control. The monoamine oxidase-A (MAO-A) gene, coding for an enzyme primarily involved in serotonin and noradrenaline catabolism, presents a well-characterized functional polymorphism consisting of a variable number of tandem repeats in the promoter region, with high-activity and low-activity variants. High-activity allele carriers have higher enzyme expression, lower amine concentration, and present higher scores on behavioral measures of impulsivity than low-activity allele carriers. Methods We studied the relationship of this polymorphism to brain activity elicited by a response inhibition task (Go/NoGo task), using blood oxygenation level–dependent (BOLD) functional magnetic resonance imaging in 24 healthy men. Results Direct comparison between groups revealed a greater BOLD response in the right ventrolateral prefrontal cortex (Brodmann’s area [BA] 45/47) in high-activity allele carriers, whereas a greater response in the right superior parietal cortex (BA 7) and bilateral extrastriate cortex (BA 18) was found in low-activity allele carriers. Conclusions These data suggest that a specific genetic variation involving serotonergic catabolism can modulate BOLD response associated with human impulsivity.

Charcot–Marie-Tooth type 1A is caused by a 1.5 Mb DNA duplication in the 17p12 chromosomal region encompassing the peripheral myelin protein 22 gene. In the present study, we compared the Real-Time PCR with the other methods currently used for the diagnosis of Charcot–Marie-Tooth. By using a combination of junction fragment PCR, analysis of microsatellite markers, and pulsed field gel electrophoresis, we identified 76 unrelated patients with 17p12 duplication. In these patients, junction fragment PCR detected 63% of cases of duplication, the microsatellite markers method revealed 74%, while the combined use of microsatellite markers and junction fragment PCR revealed 91% of cases of Charcot–Marie-Tooth type 1A. Pulsed field gel electrophoresis detected 100% of the cases with duplication, even in presence of atypical 17p12 duplication. Real-Time PCR detected 100% of the cases with Charcot–Marie-Tooth type 1A and was comparable to pulsed field gel electrophoresis. However, in contrast to pulsed field gel electrophoresis, Real-Time PCR does not need fresh blood, minimizes diagnosis time and cost, and thus can be easily used for the molecular diagnosis of Charcot–Marie-Tooth type 1A.

We describe the clinical, neuropathological and molecular findings from a patient affected with neuronal ceroid lipofuscinosis with a juvenile onset (JNCL). She was a 9-year-old right-handed girl with a normal birth and early developmental milestones. At the age of 4 the early symptoms began. Skin biopsy showed granular osmiophilic deposits (GRODs).Because JNCL with GRODs is caused by mutations in the CNL1 gene, we performed a molecular investigation by direct sequencing of nine exons of the CNL1 gene. This analysis revealed a novel mutation in homozygous form in the exon 7 that caused an aminoacid substitution at codon 222 (Leu --> Pro). Direct sequencing of the exon 7 in both parents showed the same substitution in heterozygous form.

Objective – To investigate the possible occurrence of a conversion event in three patients with adult-onset spinal muscular atrophy (SMA) type IV, which represents the mildest form within the spectrum of the SMA phenotype. Material and methods – We observed three patients with adult onset SMA and apparent isolated deletion of telomeric survival motor neuron (SMN1) exon 7. To distinguish between a deletion and a sequence conversion event of exon 7, these patients were analyzed in greater detail by a simple PCR-based assay. Results – Analysis by DdeI digestion showed products for both telomeric and centromeric copies of exon 8. These findings indicated a gene conversion event as the site for primer R111 was retained at least in one of two alleles. Conclusions – These results provide first evidence that a conversion event may be also associated with adult-onset SMA, and further support the notion that a gene conversion event is usually associated with a milder SMA phenotype and a later onset of disease.

Mutations in the SPG4 gene are the most common causes of hereditary spastic paraplegia (HSP) accounting for up to 40% of autosomal dominant (AD) forms and 12-18% of sporadic cases. The phenotype associated with HSP due to mutations in the SPG4 gene tends to be pure. There is increasing evidence, however, of patients with complicated forms of spastic paraplegia in which SPG4 mutations were identified. A cohort of 38 unrelated Italian patients with spastic paraplegia, of which 24 had a clear dominant inheritance and 14 were apparently sporadic, were screened for mutations in the SPG4 gene.We identified 11 different mutations, six of which were novel (p.Glu143GlyfsX8, p.Tyr415X, p.Asp548Asn, c.1656_1664delinsTGACCT, c.1688-3C>G and c.*2G>T) and two exon deletions previously reported. The overall rate of SPG4 gene mutation in our patients was 36.8% (14/38); in AD-HSP we observed a mutation frequency of 45.8% (11/24), in sporadic cases the frequency was 21.4% (3/14). Furthermore, we found a mutational rate of 22.2% (2/9) and 41.4% (12/29) in the complicated and pure forms, respectively. The results underlie the importance of genetic testing in all affected individuals.

TAR-DNA-binding protein 43 (TDP-43) has recently been identified as the major pathological protein in abnormal inclusions in neurons and glial cells in sporadic amyotrophic lateral sclerosis (SALS) and SOD1 negative familial cases with amyotrophic lateral sclerosis (FALS). TDP-43 is evolutionarily conserved, consisting of two RNA recognition motifs and a glycine-rich C-terminal domain. It is involved in the regulation of expression and splicing, and in other cellular processes such as microRNA biogenesis, apoptosis and cell division.1 2 Starting in early 2008, dominant mutations in TARDBP gene have been reported by several groups as a primary cause of ALS. To date, a total of 30 mutations of TARDBP have been reported not only in SOD1 -negative FALS cases (∼3%) but also in SALS cases (∼1.5%). All but one of the mutations identified (D169G) reside in exon 6 of the TARDBP gene, which encodes for the C-terminal glycine-rich domain of TDP-43. All of these mutations are dominantly inherited missense changes with the exception of a truncating mutation (Y374X) at the extreme C terminus of the protein.3 In this study, in order to investigate the presence and frequency of TARDBP mutations in a cohort of 310 south Italian patients affected by ALS, we performed a mutational screening of the exon 4 and exon 6 of the gene in SOD1 -negative FALS and SALS patients. …

To provide further insights into the distribution and spectrum of mutations at the MECP2 locus, 21 patients from Sicily were analyzed: 14 with classical RTT syndrome and seven with variant forms. Mutations in MECP2 were found in 10 of 14 classical Rett patients (71.4%), and in three of seven cases with a variant form (42.8%). Two novel mutations, not found in 100 control chromosomes, were detected.

We have identified a new neutral hemoglobin variant in a pregnant Italian woman, that resulted from a GTG--CTG replacement at codon 126 of the beta chain, corresponding to a Val--Leu amino acid change at position beta126(H4). Thermal and isopropanol stability tests were normal and there were no abnormal clinical features. Routine electrophoretic and ion exchange chromatographic methods for hemoglobin separation failed to show this variant, but reversed phase high performance liquid chromatography revealed an abnormal peak eluting near the normal beta chain. No abnormal tryptic peptide was revealed on the high performance liquid chromatographic elution pattern of the total globin digest. The mutation was determined at the DNA level by amplification of the three beta exons by polymerase chain reaction and direct sequencing of one exon that showed an abnormal migration on single strand conformational polymorphism analysis.

CADASIL (OMIM 125310) is an increasingly recognised adult-onset autosomal-dominant vascular disease that is characterised by recurrent transient ischaemic attacks and strokes (43% of patients), vascular dementia (6%), migraine with aura (40% of patients) and psychiatric disturbances (9% of patients); epilepsy has been reported in 2–10% of subjects.1 All patients revealed prominent signal abnormalities on brain magnetic resonance imaging (MRI)—leukoencephalopathy on T2- and small subcortical infarcts on T1-weighted images.2 The pathological hallmark of CADASIL is a non-amyloid and non-arteriosclerotic angiopathy, which predominantly affects the small penetrating brain arteries. Vascular lesions are characterised by degeneration and loss of smooth-muscle cells and by the presence of granular osmiophilic material (GOM) accumulating within the smooth-muscle-cell basement membrane and the surrounding extracellular matrix. Examination of several peripheral organs revealed vessel changes, including the presence of GOM deposits, providing evidence that CADASIL is a systemic arteriopathy.3 It has been reported that CADASIL is caused by single missense mutations, small in-frame deletions or splice-site mutations in the NOTCH3 gene encoding a transmembrane receptor (http://www.hgmd.cf.ac.uk/ac/gene.php?gene = NOTCH3). Almost all previously reported mutations resulted in an odd number of cysteine residues within one of the 34 epidermal growth factor (EGF)-like repeats in the …

To investigate the possible occurrence of a conversion event in three patients with adult-onset spinal muscular atrophy (SMA) type IV, which represents the mildest form within the spectrum of the SMA phenotype.We observed three patients with adult onset SMA and apparent isolated deletion of telomeric survival motor neuron (SMN1) exon 7. To distinguish between a deletion and a sequence conversion event of exon 7, these patients were analyzed in greater detail by a simple PCR-based assay.Analysis by DdeI digestion showed products for both telomeric and centromeric copies of exon 8. These findings indicated a gene conversion event as the site for primer R111 was retained at least in one of two alleles.These results provide first evidence that a conversion event may be also associated with adult-onset SMA, and further support the notion that a gene conversion event is usually associated with a milder SMA phenotype and a later onset of disease.

Hereditary spastic paraplegia (HSP) includes a group of diseases characterized by progressive spastic weakness of the lower limbs (pure forms) with possible additional signs (complicated forms). The SPG10 form is due to alteration in the kinesin1A gene (KIF5A) that encodes the neuronal kinesin heavy chain, a protein required for the anterograde axonal transport. We performed clinical, neurophysiological and molecular studies in two siblings affected by AD-HSP complicated by deafness. The screening of the KIF5A gene revealed the novel mutation p.Leu259Gln in two affected siblings and in their father with a pure form of HSP. © 2014 Elsevier B.V.

A. Magariello*, L. Citrigno*, S. Zuchner, M. Gonzalez, A. Patitucci, V. Sofia, F. L. Conforti, I. Pappalardo, R. Mazzei, C. Ungaro, M. Zappia and M. Muglia Institute of Neurological Sciences, National Research Council, Mangone (CS), Italy; Department of Human Genetics and Hussman Institute for Human Genomics, Miller School of Medicine, University of Miami, Miami, FL, USA; and Dipartimento G.F. Ingrassia Sezione di Neuroscienze Universit a di Catania, Catania, Italy

In the current study, we report a novel mutation in a large Italian pedigree from southern Italy with autosomal dominant uncomplicated spastic paraplegia. We included 35 individuals from four generations. Significant linkage to the SPG3 locus on chromosome 14 was detected with a maximum LOD score of 4.58 at D14S255. Direct sequencing of the SPG3A gene revealed a G-->A mutation at position 818 in exon 7 of the gene. This mutation created an amino-acid change from Arg to Gln at codon 217. This is the first evidence of a mutation in the SPG3A gene that alters the RD motif of atlastin. These data also confirm that mutations in the SPG3A gene are causative of AD-HSP

A large Italian pedigree from southern Italy with autosomal dominant uncomplicated spastic paraplegia is reported. The clinical picture was uniform and characterized by insidiously progressive lower extremity weakness and spasticity. The mean age at onset of symptoms was 8.3 years. Significant linkage to the SPG3 locus on chromosome 14 was detected. The authors also report their search for mutations in a gene located in the region and its exclusion as a candidate for SPG3.