@article {10520, title = {Puzzling Results from Germline Mutations Analysis in a Group of Asbestos-Exposed Patients in a High-risk Area of Northeast Italy.}, journal = {Anticancer Res}, volume = {37}, year = {2017}, month = {2017 06}, pages = {3073-3083}, abstract = {

BACKGROUND: Germline mutations of the oncosuppressor gene breast cancer 1-associated protein 1 (BAP1) were recently related to an autosomal-dominant tumor predisposition syndrome (BAP1-TPDS), characterized by uveal melanoma, malignant mesothelioma (MM), cutaneous melanoma, and other malignancies. The demonstration that BAP1 mutations are strongly associated with MM has provided a real breakthrough in the study of genetic predisposition in MM, that may explain why only a fraction of asbestos-exposed individuals go on to develop MM.

MATERIALS AND METHODS: To evaluate the possible role of BAP1 mutations in the epidemiology of sporadic MM, and their relationship with asbestos exposure, we determined the prevalence of germline BAP1 mutations by the Sanger method in a group of 29 asbestos-exposed patients, 21 of which were diagnosed with MM. They were residents of Trieste, a ship-building town in Northeast Italy with a very high incidence of mesothelioma.

RESULTS: We identified non-obviously pathogenetic germline sequence variants of BAP1 in 3/29 patients and in 2/21 MM cases (10\%).

CONCLUSION: Non obviously pathogenic germline sequence variants of BAP1 were found. Nevertheless, limitations of predictive web tools allowed us to comment on some interesting peculiarities of our findings.

}, keywords = {Aged, Aged, 80 and over, Asbestos, Environmental Exposure, Female, Germ-Line Mutation, Humans, Italy, Lung Neoplasms, Male, Mesothelioma, Middle Aged, Risk, Tumor Suppressor Proteins, Ubiquitin Thiolesterase}, issn = {1791-7530}, doi = {10.21873/anticanres.11663}, author = {Rizzardi, Clara and Athanasakis, Emmanouil and Cammisuli, Francesca and Monego, Simeone Dal and DE Spelorzi, Yeraldin Chiquinquira Castillo and Costantinides, Fulvio and Giudici, Fabiola and Pinamonti, Maurizio and Canzonieri, Vincenzo and Melato, Mauro and Pascolo, Lorella} } @article {8322, title = {Abnormal expression of leiomyoma cytoskeletal proteins involved in cell migration.}, journal = {Oncol Rep}, volume = {35}, year = {2016}, month = {2016 May}, pages = {3094-100}, abstract = {

Uterine leiomyomas are monoclonal tumors. Several factors are involved in the neoplastic transformation of the myometrium. In our study we focused on dysregulated cytoskeletal proteins in the leiomyoma as compared to the myometrium. Paired tissue samples of ten leiomyomas and adjacent myometria were obtained and analyzed by two-dimensional gel electrophoresis (2-DE). Mass spectrometry was used for protein identification, and western blotting for 2-DE data validation. The values of ten cytoskeletal proteins were found to be significantly different: eight proteins were upregulated in the leiomyoma and two proteins were downregulated. Three of the upregulated proteins (myosin regulatory light polypeptide 9, four and a half LIM domains protein 1 and LIM and SH3 domain protein 1) are involved in cell migration, while downregulated protein transgelin is involved in replicative senescence. Myosin regulatory light polypeptide 9 (MYL9) was further validated by western blotting because it is considered to be a cell migration marker in several cancers and could play a key role in leiomyoma development. Our data demonstrate significant alterations in the expression of cytoskeletal proteins involved in leiomyoma growth. A better understanding of the involvement of cytoskeletal proteins in leiomyoma pathogenesis may contribute to the identification of new therapeutic targets and the development of new pharmacological approaches.

}, issn = {1791-2431}, doi = {10.3892/or.2016.4688}, author = {Ura, Blendi and Scrimin, Federica and Arrigoni, Giorgio and Athanasakis, Emmanouil and Aloisio, Michelangelo and Monasta, Lorenzo and Ricci, Giuseppe} } @article {7774, title = {Genetic profiling of autoinflammatory disorders in patients with periodic fever: a prospective study.}, journal = {Pediatr Rheumatol Online J}, volume = {13}, year = {2015}, month = {2015}, pages = {11}, abstract = {

BACKGROUND: Periodic fever syndromes (PFS) are an emerging group of autoinflammatory disorders. Clinical overlap exists and multiple genetic analyses may be needed to assist diagnosis. We evaluated the diagnostic value of a 5-gene sequencing panel (5GP) in patients with undiagnosed PFS.

METHODS: Simultaneous double strand Sanger sequencing of MEFV, MVK, TNFRSF1A, NLRP3, NLRP12 genes was performed in 42 patients with unexplained PFS. Clinical features were correlated with genetic results.

RESULTS: None of 42 patients analyzed displayed a causative genotype. However, single or multiple genetic variants of uncertain significance were detected in 24 subjects. Only in 5 subjects a definite diagnosis was made by taking into account both genetic and clinical data (2 TRAPS syndrome; 2 FMF; 1 FCAS). Statistical analysis showed that patients carrying genetic variants in one or more of the five selected genes displayed a significantly lower response to glucocorticoids compared with subjects who had completely negative genetic results.

CONCLUSIONS: The sequencing of multiple genes is of little help in the diagnostics of PFS and can often lead to results of uncertain interpretation, thus the clinically driven sequencing of single genes should remain the recommended approach. However, the presence of single or multiple genetic variants of uncertain significance, even if not allowing any specific diagnosis, correlated with a poorer response to glucocorticoids, possibly indicating a multifactorial subgroup of PFS with differential response to pharmacological treatment.

}, keywords = {Adolescent, Carrier Proteins, Child, Cryopyrin-Associated Periodic Syndromes, Cytoskeletal Proteins, Familial Mediterranean Fever, Female, Fever, Gene Expression Profiling, Genotype, Hereditary Autoinflammatory Diseases, Humans, Intracellular Signaling Peptides and Proteins, Logistic Models, Male, Mutation, Phosphotransferases (Alcohol Group Acceptor), Prospective Studies, Receptors, Tumor Necrosis Factor, Type I, Syndrome}, issn = {1546-0096}, doi = {10.1186/s12969-015-0006-z}, author = {De Pieri, Carlo and Vuch, Josef and De Martino, Eleonora and Bianco, Anna M and Ronfani, Luca and Athanasakis, Emmanouil and Bortot, Barbara and Crovella, Sergio and Taddio, Andrea and Severini, Giovanni M and Tommasini, Alberto} } @article {3478, title = {Autosomal recessive Stickler syndrome due to a loss of function mutation in the COL9A3 gene.}, journal = {Am J Med Genet A}, volume = {164A}, year = {2014}, month = {2014 Jan}, pages = {42-7}, abstract = {

Stickler syndrome (STL) is a clinically variable and genetically heterogeneous syndrome characterized by ophthalmic, articular, orofacial, and auditory manifestations. STL has been described with both autosomal dominant and recessive inheritance. The dominant form is caused by mutations of COL2A1 (STL 1, OMIM 108300), COL11A1 (STL 2, OMIM 604841), and COL11A2 (STL 3, OMIM 184840) genes, while recessive forms have been associated with mutations of COL9A1 (OMIM 120210) and COL9A2 (OMIM 120260) genes. Type IX collagen is a heterotrimeric molecule formed by three genetically distinct chains: α1, α2, and α3 encoded by the COL9A1, COL9A2, and COL9A3 genes. Up to this time, only heterozygous mutations of COL9A3 gene have been reported in human and related to: (1) multiple epiphyseal dysplasia type 3, (2) susceptibility to an intervertebral disc disease, and (3) hearing loss. Here, we describe the first autosomal recessive Stickler family due to loss of function mutations (c.1176_1198del, p.Gln393Cysfs*25) of COL9A3 gene. These findings extend further the role of collagen genes family in the disease pathogenesis.

}, keywords = {Adolescent, Arthritis, Bone and Bones, Child, Child, Preschool, Collagen Diseases, Collagen Type IX, Connective Tissue Diseases, DNA Mutational Analysis, Facies, Female, Genes, Recessive, Hearing Loss, Hearing Loss, Sensorineural, Homozygote, Humans, Male, Mutation, Pedigree, Retinal Detachment}, issn = {1552-4833}, doi = {10.1002/ajmg.a.36165}, author = {Faletra, Flavio and d{\textquoteright}Adamo, Adamo P and Bruno, Irene and Athanasakis, Emmanouil and Biskup, Saskia and Esposito, Laura and Gasparini, Paolo} } @article {3610, title = {F402L variant in NLRP12 in subjects with undiagnosed periodic fevers and in healthy controls.}, journal = {Clin Exp Rheumatol}, volume = {32}, year = {2014}, month = {2014 Nov-Dec}, pages = {993-4}, keywords = {Cryopyrin-Associated Periodic Syndromes, Female, Humans, Intracellular Signaling Peptides and Proteins, Male, Mutation}, issn = {0392-856X}, author = {De Pieri, Carlo and Vuch, Josef and Athanasakis, Emmanouil and Severini, Giovanni Maria and Crovella, Sergio and Bianco, Anna Monica and Tommasini, Alberto} } @article {3499, title = {Next generation sequencing in nonsyndromic intellectual disability: from a negative molecular karyotype to a possible causative mutation detection.}, journal = {Am J Med Genet A}, volume = {164A}, year = {2014}, month = {2014 Jan}, pages = {170-6}, abstract = {

The identification of causes underlying intellectual disability (ID) is one of the most demanding challenges for clinical Geneticists and Researchers. Despite molecular diagnostics improvements, the vast majority of patients still remain without genetic diagnosis. Here, we report the results obtained using Whole Exome and Target Sequencing on nine patients affected by isolated ID without pathological copy number variations, which were accurately selected from an initial cohort of 236 patients. Three patterns of inheritance were used to search for: (1) de novo, (2) X-linked, and (3) autosomal recessive variants. In three of the nine proband-parent trios analyzed, we identified and validated two de novo and one X-linked potentially causative mutations located in three ID-related genes. We proposed three genes as ID candidate, carrying one de novo and three X-linked mutations. Overall, this systematic proband-parent trio approach using next generation sequencing could explain a consistent percentage of patients with isolated ID, thus increasing our knowledge on the molecular bases of this disease and opening new perspectives for a better diagnosis, counseling, and treatment.

}, keywords = {Computational Biology, Exome, Female, Genes, Recessive, Genes, X-Linked, Genome-Wide Association Study, High-Throughput Nucleotide Sequencing, Humans, Intellectual Disability, Karyotype, Male, Mutation, Workflow}, issn = {1552-4833}, doi = {10.1002/ajmg.a.36274}, author = {Athanasakis, Emmanouil and Licastro, Danilo and Faletra, Flavio and Fabretto, Antonella and Dipresa, Savina and Vozzi, Diego and Morgan, Anna and d{\textquoteright}Adamo, Adamo P and Pecile, Vanna and Biarn{\'e}s, Xevi and Gasparini, Paolo} } @article {8098, title = {The p53 transcriptional pathway is preserved in ATMmutated and NOTCH1mutated chronic lymphocytic leukemias.}, journal = {Oncotarget}, volume = {5}, year = {2014}, month = {2014 Dec 30}, pages = {12635-45}, abstract = {

By using next generation sequencing, we have analyzed 108 B chronic lymphocytic leukemia (B-CLL) patients. Among genes involved in the TP53 pathway, we found frequent mutations in ATM (n=18), TP53 (n=10) and NOTCH1 (n=10) genes, rare mutations of NOTCH2 (n=2) and CDKN1A/p21 (n=1) and no mutations in BAX, MDM2, TNFRSF10A and TNFRSF10B genes. The in vitro treatment of primary B-CLL cells with the activator of p53 Nutlin-3 induced the transcription of p53 target genes, without significant differences between the B-CLL without mutations and those harboring either ATM or NOTCH1mutations. On the other hand, the subgroup of TP53mutated B-CLL exhibited a significantly lower induction of the p53 target genes in response to Nutlin-3 as compared to the other B-CLL samples. However, among the TP53mutated B-CLL, those showing mutations in the high hot spot region of the DNA binding domain [273-280 aa] maintained a significantly higher p53-dependent transcriptional activity as compared to the other TP53mutated B-CLL samples. Since the ability to elicit a p53-dependent transcriptional activity in vitro has a positive prognostic significance, our data suggest that ATMmutated, NOTCH1mutated and surprisingly, also a subset of TP53mutated B-CLL patients might benefit from therapeutic combinations including small molecule activator of the p53 pathway.

}, keywords = {Aged, Aged, 80 and over, Ataxia Telangiectasia Mutated Proteins, Base Sequence, Female, Genes, p53, Humans, Leukemia, Lymphocytic, Chronic, B-Cell, Male, Models, Molecular, Molecular Sequence Data, Mutation, Receptor, Notch1, Signal Transduction, Tumor Suppressor Protein p53}, issn = {1949-2553}, doi = {10.18632/oncotarget.2211}, author = {Athanasakis, Emmanouil and Melloni, Elisabetta and Rigolin, Gian Matteo and Agnoletto, Chiara and Voltan, Rebecca and Vozzi, Diego and Piscianz, Elisa and Segat, Ludovica and dal Monego, Simeone and Cuneo, Antonio and Secchiero, Paola and Zauli, Giorgio} } @article {3524, title = {TNF-α SNP rs1800629 and risk of relapse in childhood acute lymphoblastic leukemia: relation to immunophenotype.}, journal = {Pharmacogenomics}, volume = {15}, year = {2014}, month = {2014 Apr}, pages = {619-27}, abstract = {

AIM: In the AIEOP-BFM ALL (Associazione Italiana Ematologia Oncologia Pediatrica-Berlin Frankfurt M{\"u}nster acute lymphoblastic leukemia) 2000 protocol, 70\% of relapsed patients had favorable prognostic features and fell within less intensive polychemotherapeutic regimens, suggesting the need for better assessing lower risk stratification.

MATERIALS \& METHODS: A novel two-phase study design selected 614 children to be genotyped for TNF-α SNP rs1800629 (-308G>A). A weighted Cox model was applied to evaluate the SNP effect on hazard of relapse, adjusting for immunophenotype, risk group, age and gender and including interaction terms.

RESULTS: Significant interaction was found with immunophenotypes (p = 0.0007, with minor allele genotypes being adverse genetic markers in B-cell acute lymphoblastic leukemia and protective ones in T-cell acute lymphoblastic leukemia), and also with risk protocols (p = 0.0041, with minor allele genotypes as prognostic factor of relapse for standard risk patients [only one T-cell acute lymphoblastic leukemia in the subgroup analyzed]).

CONCLUSION: The presence of at least one A allele in TNF-α SNP rs1800629 should suggest a closer monitoring in B-cell acute lymphoblastic leukemia standard risk patients.

}, keywords = {Adolescent, Antineoplastic Agents, Child, Child, Preschool, Drug Resistance, Neoplasm, Female, Genotype, Humans, Infant, Leukemia, Lymphocytic, Chronic, B-Cell, Male, Phenotype, Precursor Cell Lymphoblastic Leukemia-Lymphoma, Precursor T-Cell Lymphoblastic Leukemia-Lymphoma, Recurrence, Risk Assessment, Steroids, Tumor Necrosis Factor-alpha}, issn = {1744-8042}, doi = {10.2217/pgs.13.249}, author = {Franca, Raffaella and Rebora, Paola and Athanasakis, Emmanouil and Favretto, Diego and Verzegnassi, Federico and Basso, Giuseppe and Tommasini, Alberto and Valsecchi, Maria Grazia and Decorti, Giuliana and Rabusin, Marco} } @article {1956, title = {A novel CRYBB2 missense mutation causing congenital autosomal dominant cataract in an Italian family.}, journal = {Ophthalmic Genet}, volume = {34}, year = {2013}, month = {2013 Mar-Jun}, pages = {115-7}, abstract = {

Congenital cataract is a leading cause of visual impairment in children and brings approximately 10\% of childhood blindness worldwide. Molecular analysis revealed ~60 loci to be associated with several phenotypes of childhood cataracts. Until now, more than 30 loci and 18 genes on different chromosomes have been associated with autosomal dominant congenital cataract (ADCC). Here, we present a three-generation Italian family with a non syndromic ADCC. A linkage analysis carried out using HumanCytoSNP-12 DNA Analysis BeadChip led us to identify ten genomic regions virtually involved in the disease. All the genes located in these regions were scored for possible relationship with ADCC and, according to a strict clinical and genetic selection, 4 genes have been analyzed. A novel sequence variant was found in the CRYBB2 gene (p.Ser143Phe). This variant affects a conserved aminoacid in the third Greek key motif of the protein, cosegregates with the disease phenotype in all affected individuals and is not present both in the unaffected family members and 100 healthy control subjects. Finally, we identified the first CRYBB2 mutation in an Italian family causing a clinical picture of ADCC.

}, keywords = {Amino Acid Sequence, beta-Crystallin B Chain, Cataract, DNA Mutational Analysis, Female, Genes, Dominant, Genetic Linkage, Genotype, Humans, Italy, Male, Molecular Sequence Data, Mutation, Missense, Pedigree, Phenotype}, issn = {1744-5094}, doi = {10.3109/13816810.2012.707273}, author = {Faletra, Flavio and d{\textquoteright}Adamo, Adamo Pio and Pensiero, Stefano and Athanasakis, Emmanouil and Catalano, Dario and Bruno, Irene and Gasparini, Paolo} } @article {1960, title = {Genetics of food preferences: a first view from silk road populations.}, journal = {J Food Sci}, volume = {77}, year = {2012}, month = {2012 Dec}, pages = {S413-8}, abstract = {

Food preferences are the main factor driving food intake and choice. There are good reasons to suspect some genetic influence on food acceptance, not least because genetic factors are implicated in a number of factors that are likely to be related to food choice. In addition, some food dislikes show themselves early in life, before there is any evidence for aversive experiences. Although taste has been widely studied in regards of pure tastes such as bitter or sweet perception, the relationship between taste-related genes and food preferences has seldom been explored. In this work we investigated relationship of 37 taste-related genes with food preferences. The study was carried out during a scientific expedition through Caucasus and Central Asia (Silk Road) analyzing more than 400 samples from 5 different countries. A food preference questionnaire was administered to each participant and a DNA sample was obtained. Other information, such as age, sex, life style and anthropometrical measures, were also collected. We found significant associations with variants of: (1) TAS1R2 [Correction added after initial online publication on 27 Aug 2012. TAS1R3 was changed to TAS1R2.] gene and liking of Vodka (P= 1.6 {\texttimes} 10(-3)), white wine (P= 4.0 {\texttimes} 10(-4)) and lamb meat (P= 1.6 {\texttimes} 10(-3)); (2) PCLB2 gene and preference for Hot Tea (P= 8.0 {\texttimes} 10(-4)); (3) TPRV1 gene and beet liking (P= 3.8 {\texttimes} 10(-5)); and (4) ITPR3 gene and liking of both lamb meat (5.8 {\texttimes} 10(-4)) and sheep cheese (8.9{\texttimes}10(-4)). These findings give a new insight on a better understanding, of genetic factors influencing food preferences which is critical to the development of effective dietary interventions, especially for people that may be genetically not predisposed for liking specific nutrients.

}, keywords = {Adolescent, Adult, Aged, Aged, 80 and over, Azerbaijan, Child, Choice Behavior, Cohort Studies, Female, Food Habits, Food Preferences, Gene Frequency, Genotype, Georgia, Humans, Kazakhstan, Linear Models, Male, Middle Aged, Polymorphism, Single Nucleotide, Quantitative Trait Loci, Questionnaires, Tajikistan, Taste, Uzbekistan, Young Adult}, issn = {1750-3841}, doi = {10.1111/j.1750-3841.2012.02852.x}, author = {Pirastu, Nicola and Robino, Antonietta and Lanzara, Carmela and Athanasakis, Emmanouil and Esposito, Laura and Tepper, Beverly J and Gasparini, Paolo} } @article {1967, title = {Molecular diagnosis of Usher syndrome: application of two different next generation sequencing-based procedures.}, journal = {PLoS One}, volume = {7}, year = {2012}, month = {2012}, pages = {e43799}, abstract = {

Usher syndrome (USH) is a clinically and genetically heterogeneous disorder characterized by visual and hearing impairments. Clinically, it is subdivided into three subclasses with nine genes identified so far. In the present study, we investigated whether the currently available Next Generation Sequencing (NGS) technologies are already suitable for molecular diagnostics of USH. We analyzed a total of 12 patients, most of which were negative for previously described mutations in known USH genes upon primer extension-based microarray genotyping. We enriched the NGS template either by whole exome capture or by Long-PCR of the known USH genes. The main NGS sequencing platforms were used: SOLiD for whole exome sequencing, Illumina (Genome Analyzer II) and Roche 454 (GS FLX) for the Long-PCR sequencing. Long-PCR targeting was more efficient with up to 94\% of USH gene regions displaying an overall coverage higher than 25{\texttimes}, whereas whole exome sequencing yielded a similar coverage for only 50\% of those regions. Overall this integrated analysis led to the identification of 11 novel sequence variations in USH genes (2 homozygous and 9 heterozygous) out of 18 detected. However, at least two cases were not genetically solved. Our result highlights the current limitations in the diagnostic use of NGS for USH patients. The limit for whole exome sequencing is linked to the need of a strong coverage and to the correct interpretation of sequence variations with a non obvious, pathogenic role, whereas the targeted approach suffers from the high genetic heterogeneity of USH that may be also caused by the presence of additional causative genes yet to be identified.

}, keywords = {Child, Preschool, Exome, Genome, Human, High-Throughput Nucleotide Sequencing, Humans, Molecular Diagnostic Techniques, Pilot Projects, Sequence Analysis, DNA, Usher Syndromes}, issn = {1932-6203}, doi = {10.1371/journal.pone.0043799}, author = {Licastro, Danilo and Mutarelli, Margherita and Peluso, Ivana and Neveling, Kornelia and Wieskamp, Nienke and Rispoli, Rossella and Vozzi, Diego and Athanasakis, Emmanouil and D{\textquoteright}Eustacchio, Angela and Pizzo, Mariateresa and D{\textquoteright}Amico, Francesca and Ziviello, Carmela and Simonelli, Francesca and Fabretto, Antonella and Scheffer, Hans and Gasparini, Paolo and Banfi, Sandro and Nigro, Vincenzo} } @article {1703, title = {Association of a variant in the CHRNA5-A3-B4 gene cluster region to heavy smoking in the Italian population.}, journal = {Eur J Hum Genet}, volume = {19}, year = {2011}, month = {2011 May}, pages = {593-6}, abstract = {

Large-scale population studies have established that genetic factors contribute to individual differences in smoking behavior. Linkage and genome-wide association studies have shown many chromosomal regions and genes associated with different smoking behaviors. One study was the association of single-nucleotide polymorphisms (SNPs) in the CHRNA5-A3-B4 gene cluster to nicotine addiction. Here, we report a replication of this association in the Italian population represented by three genetically isolated populations. One, the Val Borbera, is a genetic isolate from North-Western Italy; the Cilento population, is located in South-Western Italy; and the Carlantino village is located in South-Eastern Italy. Owing to their position and their isolation, the three populations have a different environment, different history and genetic structure. The variant A of the rs1051730 SNP was significantly associated with smoking quantity in two populations, Val Borbera and Cilento, no association was found in Carlantino population probably because difference in LD pattern in the variant region.

}, keywords = {Genetic Predisposition to Disease, Genome-Wide Association Study, Humans, Italy, Multigene Family, Nerve Tissue Proteins, Polymorphism, Single Nucleotide, Receptors, Nicotinic, Smoking, Tobacco Use Disorder}, issn = {1476-5438}, doi = {10.1038/ejhg.2010.240}, author = {Sorice, Rossella and Bione, Silvia and Sansanelli, Serena and Ulivi, Sheila and Athanasakis, Emmanouil and Lanzara, Carmela and Nutile, Teresa and Sala, Cinzia and Camaschella, Clara and d{\textquoteright}Adamo, Pio and Gasparini, Paolo and Ciullo, Marina and Toniolo, Daniela} } @article {1779, title = {High-throughput genotyping robot-assisted method for mutation detection in patients with hypertrophic cardiomyopathy.}, journal = {Diagn Mol Pathol}, volume = {20}, year = {2011}, month = {2011 Sep}, pages = {175-9}, abstract = {

Hypertrophic cardiomyopathy (HCM) is the most frequent autosomal dominant genetic heart muscle disease and the most common cause of sudden cardiac death in young people (under 30 y of age), who are often unaware of their underlying condition. Genetic screening is now considered a fundamental tool for clinical management of HCM families. However, the high genetic heterogeneity of HCM makes genetic screening very expensive. Here, we propose a new high-throughput genotyping method based on a HCM 96-well sequencing plate for the analysis of 8 of the most frequent HCM-causing sarcomeric genes by automating several processes required for direct sequencing, using a commercially available robotic systems and routinely used instruments. To assess the efficiency of the robot-assisted method, we have analyzed the entire coding sequence and flanking intronic sequences of the 8 sarcomeric genes in samples from 18 patients affected by HCM and their relatives, which revealed 9 different mutations, 3 of which were novel. The automated, robot-assisted assembling of polymerase chain reaction, purification of polymerase chain reaction products, and assembly of sequencing reactions resulted in a substantial saving of time, reagent costs, and reduction of human errors, and can therefore be proposed as a primary strategy for mutation identification in HCM genetic screening in many medical genetic laboratories.

}, keywords = {Cardiomyopathy, Hypertrophic, DNA Mutational Analysis, Genetic Predisposition to Disease, Genetic Testing, Genotyping Techniques, High-Throughput Nucleotide Sequencing, Humans, Muscle Proteins, Mutation, Robotics}, issn = {1533-4066}, doi = {10.1097/PDM.0b013e31820b34fb}, author = {Bortot, Barbara and Athanasakis, Emmanouil and Brun, Francesca and Rizzotti, Diego and Mestroni, Luisa and Sinagra, Gianfranco and Severini, Giovanni Maria} } @article {1733, title = {Phospholipase C-β3 is a key modulator of IL-8 expression in cystic fibrosis bronchial epithelial cells.}, journal = {J Immunol}, volume = {186}, year = {2011}, month = {2011 Apr 15}, pages = {4946-58}, abstract = {

Respiratory insufficiency is the major cause of morbidity and mortality in patients affected by cystic fibrosis (CF). An excessive neutrophilic inflammation, mainly orchestrated by the release of IL-8 from bronchial epithelial cells and amplified by chronic bacterial infection with Pseudomonas aeruginosa, leads to progressive tissue destruction. The anti-inflammatory drugs presently used in CF patients have several limitations, indicating the need for identifying novel molecular targets. To address this issue, we preliminarily studied the association of 721 single nucleotide polymorphisms from 135 genes potentially involved in signal transduction implicated in neutrophil recruitment in a cohort of F508del homozygous CF patients with either severe or mild progression of lung disease. The top ranking association was found for a nonsynonymous polymorphism of the phospholipase C-β3 (PLCB3) gene. Studies in bronchial epithelial cells exposed to P. aeruginosa revealed that PLCB3 is implicated in extracellular nucleotide-dependent intracellular calcium signaling, leading to activation of the protein kinase Cα and Cβ and of the nuclear transcription factor NF-κB p65. The proinflammatory pathway regulated by PLCB3 acts by potentiating the Toll-like Receptors{\textquoteright} signaling cascade and represents an interesting molecular target to attenuate the excessive recruitment of neutrophils without completely abolishing the inflammatory response.

}, keywords = {Adenosine Triphosphate, Calcium, Cell Line, Transformed, Cystic Fibrosis, Enzyme Activation, Epithelial Cells, Gene Expression, Gene Frequency, Genotype, Green Fluorescent Proteins, Host-Pathogen Interactions, Humans, Interleukin-8, Isoenzymes, Lung Diseases, Microscopy, Fluorescence, Phospholipase C beta, Polymorphism, Single Nucleotide, Protein Kinase C, Protein Kinase C beta, Pseudomonas aeruginosa, RNA Interference, Toll-Like Receptors, Transcription Factor RelA}, issn = {1550-6606}, doi = {10.4049/jimmunol.1003535}, author = {Bezzerri, Valentino and d{\textquoteright}Adamo, Pio and Rimessi, Alessandro and Lanzara, Carmen and Crovella, Sergio and Nicolis, Elena and Tamanini, Anna and Athanasakis, Emmanouil and Tebon, Maela and Bisoffi, Giulia and Drumm, Mitchell L and Knowles, Michael R and Pinton, Paolo and Gasparini, Paolo and Berton, Giorgio and Cabrini, Giulio} } @article {1644, title = {A polymorphism in the 5{\textquoteright} UTR of the DEFB1 gene is associated with the lung phenotype in F508del homozygous Italian cystic fibrosis patients.}, journal = {Clin Chem Lab Med}, volume = {49}, year = {2011}, month = {2011 Jan}, pages = {49-54}, abstract = {

BACKGROUND: The identification of cystic fibrosis (CF) patients who are at greater risk of lung damage could be clinically valuable. Thus, we attempted to replicate previous findings and verify the possible association between three single nucleotide polymorphisms (SNPs c.-52G>A, c.-44C>G and c.-20G>A) in the 5{\textquoteright} untranslated region (5{\textquoteright} UTR) of the β defensin 1 (DEFB1) gene and the CF pulmonary phenotype.

METHODS: Genomic DNA from 92 Italian CF patients enrolled in different regional CF centres was extracted from peripheral blood and genotyped for DEFB1 SNPs using TaqMan({\textregistered}) allele specific probes. In order to avoid genetic confounding causes that can account for CF phenotype variability, all patients were homozygous for the F508del CFTR mutation, and were then classified on the basis of clinical and functional data as mild lung phenotype (Mp, n=50) or severe lung phenotype patients (Sp, n=42).

RESULTS: For the c.-20G>A SNP, the frequency of the A allele, as well as the AA genotype, were significantly more frequent in Mp than in Sp patients, and thus this was associated with a protective effect against severe pulmonary disease (OR=0.48 and 0.28, respectively). The effect of the c.-20G>A A allele is consistent with a recessive model, and the protective effect against Sp is exerted only when it is present in homozygosis. For the other two SNPs, no differences were observed as allelic and genotypic frequency in the two subgroups of CF patients.

CONCLUSIONS: Our results, although necessary to be confirmed in larger and multiethnic populations, reinforce DEFB1 as a candidate modifier gene of the CF pulmonary phenotype.

}, keywords = {5{\textquoteright} Untranslated Regions, Adult, beta-Defensins, Cystic Fibrosis, Cystic Fibrosis Transmembrane Conductance Regulator, Female, Genotype, Homozygote, Humans, Italy, Male, Mutation, Phenotype, Polymorphism, Genetic, Young Adult}, issn = {1437-4331}, doi = {10.1515/CCLM.2011.023}, author = {Crovella, Sergio and Segat, Ludovica and Amato, Annalisa and Athanasakis, Emmanouil and Bezzerri, Valentino and Braggion, Cesare and Casciaro, Rosaria and Castaldo, Giuseppe and Colombo, Carla and Covone, Angela Elvira and De Rose, Virginia and Gagliardini, Rolando and Lanzara, Carmen and Minicucci, Laura and Morgutti, Marcello and Nicolis, Elena and Pardo, Francesca and Quattrucci, Serena and Raia, Valeria and Ravazzolo, Roberto and Seia, Manuela and Stanzial, Valentino and Termini, Lisa and Zazzeron, Laura and Cabrini, Giulio and Gasparini, Paolo} } @article {1787, title = {Vertebral defects in patients with Peters plus syndrome and mutations in B3GALTL.}, journal = {Ophthalmic Genet}, volume = {32}, year = {2011}, month = {2011 Nov}, pages = {256-8}, keywords = {Abnormalities, Multiple, Alternative Splicing, Child, Preschool, Cleft Lip, Consanguinity, Cornea, Female, Galactosyltransferases, Glucosyltransferases, Growth Disorders, Humans, Limb Deformities, Congenital, Point Mutation, Spine}, issn = {1744-5094}, doi = {10.3109/13816810.2011.587082}, author = {Faletra, Flavio and Athanasakis, Emmanouil and Minen, Federico and Fornasier, Federico and Marchetti, Federico and Gasparini, Paolo} } @article {1689, title = {A 3{\textquoteright}UTR SNP in NLRP3 gene is associated with susceptibility to HIV-1 infection.}, journal = {J Acquir Immune Defic Syndr}, volume = {54}, year = {2010}, month = {2010 Jul}, pages = {236-40}, abstract = {

OBJECTIVES: Innate immunity genes polymorphisms are known to be involved in the multifactorial susceptibility to HIV-1 infection. Recently it has been hypothesized that inflammasomes could play an important role in the host response to viruses. The aim of our study is to verify if single-nucleotide polymorphisms (SNPs) in genes encoding for NALPs-innate immune receptors that form molecular complexes leading to the production of IL-1beta and the activation of immune response-could influence the individual susceptibility to HIV-1.

DESIGN: We performed an association study analyzing 2 NLRP1 and NLRP3 SNPs in HIV-1 vertically infected Brazilian children (n = 135), HIV-1-infected Brazilain adults (n = 192) and HIV-1-positive Italian seropositive subjects (n = 192).

RESULTS: The 3{\textquoteright}UTR NLRP3 rs10754558 SNP was associated with HIV-1 infection in all the studied groups. The frequency of rs10754558 G allele was differently distributed within seropositive subjects (HIV+) and controls, and in particular the GG genotype was less frequent in HIV+.

CONCLUSIONS: susceptibility to HIV-1 infection is associated with a 3{\textquoteright}UTR NLRP3 polymorphism. This is the first report linking SNPs in the NALPs with HIV-1 infection, and further epidemiologic and functional studies are needed to deeper investigate the role of inflammasome in the susceptibility to HIV-1 infection.

}, keywords = {Adult, Brazil, Carrier Proteins, Case-Control Studies, Child, Child, Preschool, Female, Genetic Predisposition to Disease, Genotype, HIV Infections, HIV-1, Humans, Infant, Infectious Disease Transmission, Vertical, Male, Middle Aged, Polymorphism, Single Nucleotide, Pregnancy, Young Adult}, issn = {1944-7884}, doi = {10.1097/QAI.0b013e3181dd17d4}, author = {Pontillo, Alessandra and Brand{\~a}o, Lucas A and Guimar{\~a}es, Rafael L and Segat, Ludovica and Athanasakis, Emmanouil and Crovella, Sergio} }