Biotechnology Research Laboratory

Pavia, Italy

Biotechnology Research Laboratory

Pavia, Italy
SEARCH FILTERS
Time filter
Source Type

Desbats M.A.,University of Padua | Vetro A.,Biotechnology Research Laboratory | Limongelli I.,University of Pavia | Lunardi G.,University of Padua | And 15 more authors.
European Journal of Human Genetics | Year: 2015

Coenzyme Q 10 deficiency is a clinically and genetically heterogeneous disorder, with manifestations that may range from fatal neonatal multisystem failure, to adult-onset encephalopathy. We report a patient who presented at birth with severe lactic acidosis, proteinuria, dicarboxylic aciduria, and hepatic insufficiency. She also had dilation of left ventricle on echocardiography. Her neurological condition rapidly worsened and despite aggressive care she died at 23 h of life. Muscle histology displayed lipid accumulation. Electron microscopy showed markedly swollen mitochondria with fragmented cristae. Respiratory-chain enzymatic assays showed a reduction of combined activities of complex I+III and II+III with normal activities of isolated complexes. The defect was confirmed in fibroblasts, where it could be rescued by supplementing the culture medium with 10 μM coenzyme Q 10. Coenzyme Q 10 levels were reduced (28% of controls) in these cells. We performed exome sequencing and focused the analysis on genes involved in coenzyme Q 10 biosynthesis. The patient harbored a homozygous c.545T>G, p.(Met182Arg) alteration in COQ2, which was validated by functional complementation in yeast. In this case the biochemical and morphological features were essential to direct the genetic diagnosis. The parents had another pregnancy after the biochemical diagnosis was established, but before the identification of the genetic defect. Because of the potentially high recurrence risk, and given the importance of early CoQ 10 supplementation, we decided to treat with CoQ 10 the newborn child pending the results of the biochemical assays. Clinicians should consider a similar management in siblings of patients with CoQ10 deficiency without a genetic diagnosis. © 2015 Macmillan Publishers Limited All rights reserved.


Vetro A.,Biotechnology Research Laboratory | Pagani S.,University of Pavia | Silengo M.,University of Turin | Severino M.,Instituto Giannina Gaslini | And 4 more authors.
Molecular Cytogenetics | Year: 2014

We report on a male child ascertained at 4.8 years of age with severe growth failure, growth hormone (GH) deficiency, psychomotor delay with prevalent speech impairment, and a distinct phenotype. An evaluation of his hypothalamic-pituitary region by Magnetic Resonance Imaging (MRI) revealed pituitary hypoplasia with pituitary stalk interruption and ectopic posterior pituitary lobe, which are considered prognostic markers of permanent GH deficiency. Prenatal chromosome analysis because of increased nuchal translucency revealed a normal male karyotype, whereas postnatal high resolution banding raised the suspicion of a 2q abnormality. Subsequently, array Comparative Genomic Hybridization (array-CGH) revealed a de novo complex genomic rearrangement consisting of a 2p25 duplication and a 2q37 deletion: arr[hg19] 2p25.3p25.1(30,341-9,588,369)x3,2q37.2q37.3(235,744,424-243,041,305)x1. FISH analysis showed that the abnormal chromosome 2 mimicked the derivative of an inversion with the duplicated 2p region located distally at 2q. This is, to the best of our knowledge, the first case with distal 2p25 duplication and 2q37 deletion and pituitary malformation leading to GH deficiency. © 2014Vetro et al.; licensee BioMed Central Ltd.


PubMed | University of Turin, University of Pavia, IRCCS Ospedale Pediatrico Bambino Gesu, Instituto Giannina Gaslini and Biotechnology Research Laboratory
Type: | Journal: Molecular cytogenetics | Year: 2014

We report on a male child ascertained at 4.8years of age with severe growth failure, growth hormone (GH) deficiency, psychomotor delay with prevalent speech impairment, and a distinct phenotype. An evaluation of his hypothalamic-pituitary region by Magnetic Resonance Imaging (MRI) revealed pituitary hypoplasia with pituitary stalk interruption and ectopic posterior pituitary lobe, which are considered prognostic markers of permanent GH deficiency. Prenatal chromosome analysis because of increased nuchal translucency revealed a normal male karyotype, whereas postnatal high resolution banding raised the suspicion of a 2q abnormality. Subsequently, array Comparative Genomic Hybridization (array-CGH) revealed a de novo complex genomic rearrangement consisting of a 2p25 duplication and a 2q37 deletion: arr[hg19] 2p25.3p25.1(30,341-9,588,369)x3,2q37.2q37.3(235,744,424-243,041,305)x1. FISH analysis showed that the abnormal chromosome 2 mimicked the derivative of an inversion with the duplicated 2p region located distally at 2q. This is, to the best of our knowledge, the first case with distal 2p25 duplication and 2q37 deletion and pituitary malformation leading to GH deficiency.


Bersano A.,Irccs C Mondino National Neurological Institute Foundation | Bersano A.,Irccs Foundation Neurological Institute Cbesta | Zuffardi O.,University of Pavia | Pantoni L.,Stroke Unit and Neurology | And 6 more authors.
Journal of Stroke and Cerebrovascular Diseases | Year: 2015

Background: The pathogenesis of cerebral small-vessel disease (SVD) is still incompletely understood, although evidence from family and twin studies supports the hypothesis that genetic factors may contribute to SVD pathogenesis. Identification of genetic susceptibility factors for SVD may improve our knowledge on SVD pathogenesis. SVE-LA (Small Vessel and Lacunar) project is a multicenter prospective Lombardia region study aimed at applying innovative genetic technologies and accurate patient phenotyping to discover the genetic basis of SVD. Methods: A continuous series of subjects (aged 15-80 years) with a clinically and radiologically defined lacunar stroke referring to the participating Lombardia region stroke centers and an adequate number of age- and sex-matched controls are being included into the study. For each patient, clinical, demographic, instrumental, and familial data are collected applying standardized forms. After informed consent, a DNA sample for genetic analysis from patients and controls has been collected. The next generation sequencing (NGS) technology was applied to systematically screen patients for the most important genetic factors both monogenic and polygenic associated with SVD. The study includes also a centralized quantitative and qualitative analysis of neuroimaging studies. Results: Between March 2011 and October 2013, 212 lacunar stroke patients and 78 controls have been collected. Mean age of cases was 65.8 ± 11.1 years and 67% were men. Conclusions: This is the first study applying systematically NGS technology on a wide series of lacunar stroke patients. A translational approach combining a systematic genetic screening with a detailed phenotyping may facilitate the discovery of genetic basis and improve our knowledge in the pathogenesis of SVD. © 2015 by National Stroke Association.


Quadri M.,University of Pavia | Quadri M.,Erasmus University Rotterdam | Vetro A.,Biotechnology Research Laboratory | Gismondi V.,Unit of Hereditary Cancer | And 6 more authors.
Familial Cancer | Year: 2015

Familial adenomatous polyposis (FAP) is a dominantly inherited syndrome leading to the development of multiple intestinal polyps and colorectal cancer. FAP is associated with germline defects of APC tumor suppressor gene; although truncating mutations account for the majority of cases, large APC deletions represent a common disease-causing defect. While a number of intragenic deletions have been well-characterized, sequencing data of breakpoints involved in large APC rearrangements are extremely scanty. We characterized six deletions identified by multiplex ligation-dependent probe amplification (three intragenic and three larger deletions encompassing the APC locus): in each case, we precisely mapped the breakpoints by array-comparative genomic hybridization and/or long-range PCR followed by sequencing. All rearrangements were novel and no rearrangements proved to be recurrent or clustered. The three intragenic deletions involved exons 4, 9 and 14, respectively; larger deletions (30,444, 265,471 and 921,295 bp in length) involved APC as well as adjacent genes. Nine out of 12 breakpoints fell within repetitive elements (5 Alu, 2 LINE, 1 Tigger and 1 MIR), while the remaining 3 fell within unique sequences. In five out of six patients, non-allelic homologous recombination or non-homologous end joining appear as the most likely mechanisms behind APC rearrangements. Although a certain variability of clinical features was detectable both between and within families with deletions, all deletion carriers were classifiable as FAP patients showing colonic and extracolonic manifestations that belong to the spectrum of the syndrome. Therefore, different sized deletions, variable breakpoint localizations and haploinsufficiency for other genes besides APC, resulted in the same FAP clinical phenotype. © 2014, Springer Science+Business Media Dordrecht.


PubMed | University of Pavia, Irccs C Mondino National Neurological Institute Foundation, Irccs Foundation Neurological Institute Cbesta, Stroke Unit and Neurology and Biotechnology Research Laboratory
Type: Journal Article | Journal: Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association | Year: 2015

The pathogenesis of cerebral small-vessel disease (SVD) is still incompletely understood, although evidence from family and twin studies supports the hypothesis that genetic factors may contribute to SVD pathogenesis. Identification of genetic susceptibility factors for SVD may improve our knowledge on SVD pathogenesis. SVE-LA (Small Vessel and Lacunar) project is a multicenter prospective Lombardia region study aimed at applying innovative genetic technologies and accurate patient phenotyping to discover the genetic basis of SVD.A continuous series of subjects (aged 15-80years) with a clinically and radiologically defined lacunar stroke referring to the participating Lombardia region stroke centers and an adequate number of age- and sex-matched controls are being included into the study. For each patient, clinical, demographic, instrumental, and familial data are collected applying standardized forms. After informed consent, a DNA sample for genetic analysis from patients and controls has been collected. The next generation sequencing (NGS) technology was applied to systematically screen patients for the most important genetic factors both monogenic and polygenic associated with SVD. The study includes also a centralized quantitative and qualitative analysis of neuroimaging studies.Between March 2011 and October 2013, 212 lacunar stroke patients and 78 controls have been collected. Mean age of cases was 65.811.1 years and 67% were men.This is the first study applying systematically NGS technology on a wide series of lacunar stroke patients. A translational approach combining a systematic genetic screening with a detailed phenotyping may facilitate the discovery of genetic basis and improve our knowledge in the pathogenesis of SVD.


Negruk V.,Biotechnology Research Laboratory
Frontiers in Plant Science | Year: 2013

The number of plant mitochondrial genomes sequenced exceeds two dozen. However, for a detailed comparative study of different phylogenetic branches more plant mitochondrial genomes should be sequenced. This article presents sequencing data and comparative analysis of mitochondrial DNA (mtDNA of the legume Vicia faba. The size of the V. faba circular mitochondrial master chromosome of cultivar Broad Windsor was estimated as 588,000 bp with a genome complexity of 387,745 bp and 52 conservative mitochondrial genes; 32 of them encoding proteins, 3 rRNA, and 17 tRNA genes. Six tRNA genes were highly homologous to chloroplast genome sequences. In addition to the 52 conservative genes, 114 unique open reading frames (ORFs were found, 36 without significant homology to any known proteins and 29 with homology to the Medicago truncatula nuclear genome and to other plant mitochondrial ORFs, 49 ORFs were not homologous to M. truncatula but possessed sequences with significant homology to other plant mitochondrial or nuclear ORFs. In general, the unique ORFs revealed very low homology to known closely related legumes, but several sequence homologies were found between V. faba, Beta vulgaris, Nicotiana tabacum, Vitis vinifera, and even the monocots Oryza sativa and Zea mays. Most likely these ORFs arose independently during angiosperm evolution (Kubo and Mikami, 2007; Kubo and Newton, 2008. Computational analysis revealed in total about 45% of V. faba mtDNA sequence being homologous to the Medicago truncatula nuclear genome (more than to any sequenced plant mitochondrial genome, and 35% of this homology ranging from a few dozen to 12,806 bp are located on chromosome 1. Apparently, mitochondrial rrn5, rrn18, rps10, ATP synthase subunit alpha, cox2, and tRNA sequences are part of transcribed nuclear mosaic ORFs. © 2013 Negruk.


Pawar H.P.,Biotechnology Research Laboratory | Lingojwar D.P.,Biotechnology Research Laboratory | Wadhai V.S.,Biotechnology Research Laboratory
Asian Journal of Microbiology, Biotechnology and Environmental Sciences | Year: 2012

Computer hardware may act as a reservoir for microorganisms and may contribute its transfer via user. This research seeks to investigate what kinds of microbes, especially aerobic bacteria that actually can contaminate computer hardware. Bacterial 16S rRNA genes generally contain nine "hypervariable regions" (V1 - V9) that demonstrate considerable sequence diversity among different bacterial species and can be used for species identification. We identified the unknown bacteria isolated from computer hardware by molecular detection and sequence analysis of 16S rRNA hypervariable region V4.The gene sequence obtained was studied in the region from V3-V6. Sequence similarity dendograms were created for hypervariable regions V3-V6. After complete analysis and using bioinformatics tools our investigation demonstrated that the V4 (nucleotide 576-682) contains diverse character and comparatively less distinguish than V3 and V6 for the Bacillus subtilis species molecular identification. © Global Science Publications.


Decio A.,University of Pavia | Tonduti D.,University of Pavia | Pichiecchio A.,C Mondino National Neurological Institute | Vetro A.,Biotechnology Research Laboratory | And 9 more authors.
American Journal of Medical Genetics, Part A | Year: 2015

COL4A1 is located in humans on chromosome13q34 and it encodes the alpha 1 chain of type IV collagen, a component of basal membrane. It is expressed mainly in the brain, muscles, kidneys and eyes. Different COL4A1 mutations have been reported in many patients who present a very wide spectrum of clinical symptoms. They typically show a multisystemic phenotype. Here we report on the case of a patient carrying a novel de novo splicing mutation of COL4A1 associated with a distinctive clinical picture characterized by onset in infancy and an unusual evolution of the neuroradiological features. At three months of age, the child was diagnosed with a congenital cataract, while his brain MRI was normal. Over the following years, the patient developed focal epilepsy, mild diplegia, asymptomatic microhematuria, raised creatine kinase levels, MRI white matter abnormalities and brain calcification on CT. During the neuroradiological follow-up the extension and intensity of the brain lesions progressively decreased. The significance of a second variant in COL4A1 carried by the child and inherited from his father remains to be clarified. In conclusion, our patient shows new aspects of this collagenopathy and possibly a COL4A1 compound heterozygosity. © 2015 Wiley Periodicals, Inc.


Birjandi N.,Tarbiat Modares University | Younesi H.,Tarbiat Modares University | Ghoreyshi A.A.,Biotechnology Research Laboratory | Rahimnejad M.,Biotechnology Research Laboratory
Journal of Chemical Technology and Biotechnology | Year: 2015

BACKGROUND: In the present study, the performance of ethanol fermentation by Saccharomyces cerevisiae and glucose degradation in a bio-electro-Fenton (BEF) system was investigated. This system contained a dual-chamber microbial fuel cell (MFC) with Nafion® membrane as a separator and is equipped with an Fe@Fe2O3/graphite electrode. The performance of MFCs under different operational conditions was evaluated in terms of glucose removal, ethanol production and electrical power output. RESULTS: The maximum ethanol production in an anaerobic condition was 11.52 g L-1. The glucose removal was 34.15 and 68.81% and the measured maximum power densities were 13.51 and 30.46 mW m-2 with graphite and Fe@Fe2O3/graphite systems, respectively. CONCLUSION: The energy production, glucose degradation and fermentation process for ethanol production from glucose by S. cerevisiae was successfully carried out in a BEF system. This study demonstrates that the BEF system, without the requirement for an external electrical energy supply and any addition of H2O2, is an effective and economical system for wastewater treatment. © 2015 Society of Chemical Industry

Loading Biotechnology Research Laboratory collaborators
Loading Biotechnology Research Laboratory collaborators