Publications by authors named "Anne Picard"

27 Publications

  • Page 1 of 1

Kinase inhibition of G2019S-LRRK2 enhances autolysosome formation and function to reduce endogenous alpha-synuclein intracellular inclusions.

Cell Death Discov 2020 8;6:45. Epub 2020 Jun 8.

Institute for Biomedicine, Eurac Research, Affiliated Institute of the University of Lübeck - Via Galvani 31, 39100 Bolzano, Italy.

The Parkinson's disease (PD)-associated kinase Leucine-Rich Repeat Kinase 2 (LRRK2) is a crucial modulator of the autophagy-lysosome pathway, but unclarity exists on the precise mechanics of its role and the direction of this modulation. In particular, LRRK2 is involved in the degradation of pathological alpha-synuclein, with pathogenic mutations precipitating neuropathology in cellular and animal models of PD, and a significant proportion of LRRK2 patients presenting Lewy neuropathology. Defects in autophagic processing and lysosomal degradation of alpha-synuclein have been postulated to underlie its accumulation and onset of neuropathology. Thus, it is critical to obtain a comprehensive knowledge on LRRK2-associated pathology. Here, we investigated a G2019S-LRRK2 recombinant cell line exhibiting accumulation of endogenous, phosphorylated alpha-synuclein. We found that G2019S-LRRK2 leads to accumulation of LC3 and abnormalities in lysosome morphology and proteolytic activity in a kinase-dependent fashion, but independent from constitutively active Rab10. Notably, LRRK2 inhibition was ineffective upon upstream blockade of autophagosome-lysosome fusion events, highlighting this step as critical for alpha-synuclein clearance.
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http://dx.doi.org/10.1038/s41420-020-0279-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7280235PMC
June 2020

Microbiota, type 2 diabetes and non-alcoholic fatty liver disease: protocol of an observational study.

J Transl Med 2019 12 4;17(1):408. Epub 2019 Dec 4.

Institute for Biomedicine (affiliated to the University of Lübeck), Eurac Research, 39100, Bolzano, Italy.

Background: Non-alcoholic fatty liver disease (NAFLD) is characterized by triglyceride accumulation in the hepatocytes in the absence of alcohol overconsumption, commonly associated with insulin resistance and obesity. Both NAFLD and type 2 diabetes (T2D) are characterized by an altered microbiota composition, however the role of the microbiota in NAFLD and T2D is not well understood. To assess the relationship between alteration in the microbiota and NAFLD while dissecting the role of T2D, we established a nested study on T2D and non-T2D individuals within the Cooperative Health Research In South Tyrol (CHRIS) study, called the CHRIS-NAFLD study. Here, we present the study protocol along with baseline and follow-up characteristics of study participants.

Methods: Among the first 4979 CHRIS study participants, 227 individuals with T2D were identified and recalled, along with 227 age- and sex-matched non-T2D individuals. Participants underwent ultrasound and transient elastography examination to evaluate the presence of hepatic steatosis and liver stiffness. Additionally, sampling of saliva and faeces, biochemical measurements and clinical interviews were carried out.

Results: We recruited 173 T2D and 183 non-T2D participants (78% overall response rate). Hepatic steatosis was more common in T2D (63.7%) than non-T2D (36.3%) participants. T2D participants also had higher levels of liver stiffness (median 4.8 kPa, interquartile range (IQR) 3.7, 5.9) than non-T2D participants (median 3.9 kPa, IQR 3.3, 5.1). The non-invasive scoring systems like the NAFLD fibrosis score (NFS) suggests an increased liver fibrosis in T2D (mean - 0.55, standard deviation, SD, 1.30) than non-T2D participants (mean - 1.30, SD, 1.17).

Discussion: Given the comprehensive biochemical and clinical characterization of study participants, once the bioinformatics classification of the microbiota will be completed, the CHRIS-NAFLD study will become a useful resource to further our understanding of the relationship between microbiota, T2D and NAFLD.
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http://dx.doi.org/10.1186/s12967-019-02130-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6891972PMC
December 2019

Application of CRISPR/Cas9 editing and digital droplet PCR in human iPSCs to generate novel knock-in reporter lines to visualize dopaminergic neurons.

Stem Cell Res 2019 12 9;41:101656. Epub 2019 Nov 9.

Institute for Biomedicine, Eurac Research, Affiliated Institute of the University of Lübeck, Bolzano, Italy. Electronic address:

Human induced pluripotent stem cells (hiPSCs) have become indispensable for disease modelling. They are an important resource to access patient cells harbouring disease-causing mutations. Derivation of midbrain dopaminergic (DAergic) neurons from hiPSCs of PD patients represents the only option to model physiological processes in a cell type that is not otherwise accessible from human patients. However, differentiation does not produce a homogenous population of DA neurons and contaminant cell types may interfere with the readout of the in vitro system. Here, we use CRISPR/Cas9 to generate novel knock-in reporter lines for DA neurons, engineered with an endogenous fluorescent tyrosine hydroxylase - enhanced green fluorescent protein (TH-eGFP) reporter. We present a reproducible knock-in strategy combined with a highly specific homologous directed repair (HDR) screening approach using digital droplet PCR (ddPCR). The knock-in cell lines that we created show a functioning fluorescent reporter system for DA neurons that are identifiable by flow cytometry.
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http://dx.doi.org/10.1016/j.scr.2019.101656DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7322529PMC
December 2019

Generation of an induced pluripotent stem cell line (EURACi005-A) from a Parkinson's disease patient carrying a homozygous exon 3 deletion in the PRKNgene.

Stem Cell Res 2019 12 25;41:101624. Epub 2019 Oct 25.

Institute for Biomedicine, Eurac Research, via Galvani 31, Bolzano 39100, Italy.

Mutations in the PRKN gene, encoding parkin, are the most frequent known cause of recessive Parkinson's disease (PD). We report the generation of an induced pluripotent stem cell (iPSC) line of a patient carrying a homozygous deletion of exon 3 in the PRKN gene. Skin fibroblasts were reprogrammed using non-integrating episomal plasmids. The generated cell line (EURACi005-A; iPS-2011) exhibits expression of pluripotency markers, the potential to differentiate into all three germ layers, and a stable karyotype. This iPSC line provides a valuable resource for further research on the pathomechanism and drug testing for PRKN-linked PD.
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http://dx.doi.org/10.1016/j.scr.2019.101624DOI Listing
December 2019

Compound heterozygous SZT2 mutations in two siblings with early-onset epilepsy, intellectual disability and macrocephaly.

Seizure 2019 Mar 23;66:81-85. Epub 2018 Dec 23.

Institute for Biomedicine, Eurac Research, Affiliated Institute of the University of Lübeck, Bolzano, Italy; Department of Neurology, University of Lübeck, Lübeck, Germany.

Purpose: Mutations in SZT2 have been previously reported in several cases of early onset epilepsy and intellectual disability. In this study we investigate potential causal mutations in two male siblings affected by early onset epilepsy, intellectual disability and macrocephaly.

Methods: We use family-based whole-exome sequencing to identify candidate variants.

Results: We report the identification of two potential causal SZT2 mutations in compound heterozygous state. We observe considerable differences in the clinical phenotype severity of the two affected individuals. The cerebral MRI revealed no abnormalities in the older affected brother, while in the youngest one it revealed a right frontal polymicrogiria. Moreover, while good seizure control was achieved in the older affected individual the younger brother is affected by pharmacoresistant epilepsy, progressive spastic paraplegia, cortical myoclonus and a more severe intellectual disability. We also analyzed the relative location of the reported pathogenic mutations in the SZT2 protein.

Conclusion: Variable phenotypic expressivity is observed for this condition, while the location and type of mutations in SZT2 also has a potential impact on epilepsy severity. These findings extend our knowledge of epileptogenic conditions related to SZT2 and mTOR signaling.
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http://dx.doi.org/10.1016/j.seizure.2018.12.021DOI Listing
March 2019

Exploring digenic inheritance in arrhythmogenic cardiomyopathy.

BMC Med Genet 2017 12 8;18(1):145. Epub 2017 Dec 8.

Institute for Biomedicine, Eurac Research, Affiliated Institute of the University of Lübeck, Bolzano, Italy.

Background: Arrhythmogenic cardiomyopathy (ACM) is an inherited genetic disorder, characterized by the substitution of heart muscle with fibro-fatty tissue and severe ventricular arrhythmias, often leading to heart failure and sudden cardiac death. ACM is considered a monogenic disorder, but the low penetrance of mutations identified in patients suggests the involvement of additional genetic or environmental factors.

Methods: We used whole exome sequencing to investigate digenic inheritance in two ACM families where previous diagnostic tests have revealed a PKP2 mutation in all affected and some healthy individuals. In family members with PKP2 mutations we determined all genes that harbor variants in affected but not in healthy carriers or vice versa. We computationally prioritized the most likely candidates, focusing on known ACM genes and genes related to PKP2 through protein interactions, functional relationships, or shared biological processes.

Results: We identified four candidate genes in family 1, namely DAG1, DAB2IP, CTBP2 and TCF25, and eleven candidate genes in family 2. The most promising gene in the second family is TTN, a gene previously associated with ACM, in which the affected individual harbors two rare deleterious-predicted missense variants, one of which is located in the protein's only serine kinase domain.

Conclusions: In this study we report genes that might act as digenic players in ACM pathogenesis, on the basis of co-segregation with PKP2 mutations. Validation in larger cohorts is still required to prove the utility of this model.
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http://dx.doi.org/10.1186/s12881-017-0503-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5723071PMC
December 2017

Plasma and White Blood Cells Show Different miRNA Expression Profiles in Parkinson's Disease.

J Mol Neurosci 2017 Jun 24;62(2):244-254. Epub 2017 May 24.

Institute for Biomedicine, Eurac Research, Affiliated Institute of the University of Lübeck, 39100, Bolzano, Italy.

Parkinson's disease (PD) diagnosis is based on the assessment of motor symptoms, which manifest when more than 50% of dopaminergic neurons are degenerated. To date, no validated biomarkers are available for the diagnosis of PD. The aims of the present study are to evaluate whether plasma and white blood cells (WBCs) are interchangeable biomarker sources and to identify circulating plasma-based microRNA (miRNA) biomarkers for an early detection of PD. We profiled plasma miRNA levels in 99 L-dopa-treated PD patients from two independent data collections, in ten drug-naïve PD patients, and in unaffected controls matched by sex and age. We evaluated expression levels by reverse transcription and quantitative real-time PCR (RT-qPCR) and combined the results from treated PD patients using a fixed effect inverse-variance weighted meta-analysis. We revealed different expression profiles comparing plasma and WBCs and drug-naïve and L-dopa-treated PD patients. We observed an upregulation trend for miR-30a-5p in L-dopa-treated PD patients and investigated candidate target genes by integrated in silico analyses. We could not analyse miR-29b-3p, normally expressed in WBCs, due to the very low expression in plasma. We observed different expression profiles in WBCs and plasma, suggesting that they are both suitable but not interchangeable peripheral sources for biomarkers. We revealed miR-30a-5p as a potential biomarker for PD in plasma. In silico analyses suggest that miR-30a-5p might have a regulatory role in mitochondrial dynamics and autophagy. Further investigations are needed to confirm miR-30a-5p deregulation and targets and to investigate the influence of L-dopa treatment on miRNA expression levels.
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http://dx.doi.org/10.1007/s12031-017-0926-9DOI Listing
June 2017

Elevated levels of alpha-synuclein blunt cellular signal transduction downstream of Gq protein-coupled receptors.

Cell Signal 2017 01 18;30:82-91. Epub 2016 Nov 18.

Center for Biomedicine, EURAC Research, Via Galvani 31, Bolzano 39100, Italy(1). Electronic address:

Alpha-synuclein is central to Parkinson's disease pathogenesis and pathology, however its precise functions are still unclear. It has been shown to bind both PLCβ1 and MAPKs, but how this property influences the downstream signaling of Gq protein-coupled receptors has not been elucidated. Here we show that recombinant expression of alpha-synuclein in human neuroblastoma cells enhances cellular levels of PLCβ1 but blunts its signaling pathway, preventing the agonist-dependent rise of cytoplasmic Ca. In addition, overexpressing alpha-synuclein abolishes the activation of ERK1/2 upon agonist stimulation, indicating an upstream action in the signal transduction pathway. This data demonstrates that alpha-synuclein, when recombinantly expressed, interferes with the normal signaling of Gq-protein coupled receptors, which are then dysfunctional. Since many neurotransmitter systems utilize these receptor signaling pathways to mediate different abilities affected in Parkinson's disease, we argue this novel perspective might be helpful in designing treatment strategies for some of the non-motor symptoms in Parkinson's disease and synucleinopathies.
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http://dx.doi.org/10.1016/j.cellsig.2016.11.012DOI Listing
January 2017

Design and Synthesis of Selurampanel, a Novel Orally Active and Competitive AMPA Receptor Antagonist.

ChemMedChem 2017 02 15;12(3):197-201. Epub 2016 Nov 15.

Global Discovery Chemistry, Novartis Institute for Biomedical Research, 4002, Basel, Switzerland.

A series of potent quinazolinedione sulfonamide antagonists of the α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptor were designed and synthesized. The structure-activity relationships (SAR) and in vivo activity of the series were investigated. In particular, compound 1 S (selurampanel; N-[7-isopropyl-6-(2-methylpyrazol-3-yl)-2,4-dioxo-1H-quinazolin-3-yl]methanesulfonamide) has shown excellent oral potency against maximal electroshock seizure (MES)-induced generalized tonic-clonic seizures in rodents as well as significant activity in patients suffering from various forms of epilepsy. The X-ray crystal structure of selurampanel bound to the AMPA receptor hGluA was also obtained.
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http://dx.doi.org/10.1002/cmdc.201600467DOI Listing
February 2017

Overexpression of blood microRNAs 103a, 30b, and 29a in L-dopa-treated patients with PD.

Neurology 2015 Feb 16;84(7):645-53. Epub 2015 Jan 16.

From the Center for Biomedicine (A.S., L.F., S.Z., H.B., A.P., A.Z., G.G., I.P., M.F.F., P.P.P., A.A.H., F.S.D., C.S.), European Academy Bozen/Bolzano (EURAC), Bolzano, Italy, affiliated institute of the University of Lübeck, Germany; Department of Neurology (S.Z., P.P.P.), General Central Hospital, Bolzano; IRCCS Institute of Neurological Sciences of Bologna (P.C.); Department of Biomedical and NeuroMotor Sciences (P.C.), Alma Mater Studiorum-University of Bologna, Italy; Department of Neurology (P.P.P.), University of Lübeck, Germany.

Objective: The aims of the present study were to profile the expression of several candidate microRNAs (miRNAs) in blood from L-dopa-treated and drug-naive patients with Parkinson disease (PD) vs unaffected controls and to interpret the miRNA expression data in a biological context.

Methods: We analyzed RNAs from peripheral blood of 36 L-dopa-treated, 10 drug-naive patients with PD and unaffected controls matched 1:1 by sex and age. We evaluated expression by reverse transcription-quantitative real-time PCR, and we analyzed data using a 2-tailed paired t test. To detect miRNA targets, several miRNA resources were combined to generate an overall score for each candidate gene using weighted rank aggregation.

Results: Significant overexpression of miR-103a-3p (p < 0.0001), miR-30b-5p (p = 0.002), and miR-29a-3p (p = 0.005) in treated patients with PD was observed, and promising candidate target genes for these were revealed by an integrated in silico analysis.

Conclusions: We revealed 3 candidate biomarkers for PD. miRNAs 30b-5p and 29a-3p replicated a documented deregulation in PD albeit opposite to published data, while for miR-103a-3p, we demonstrated for the first time an overexpression in treated patients with PD. Expression studies in patients and/or in isolated peripheral blood mononuclear cells before and after L-dopa administration are necessary to define the involvement of L-dopa treatment in the observed overexpression. Our in silico analysis to prioritize targets of deregulated miRNAs identified candidate target genes, including genes related to neurodegeneration and PD. Despite the preliminary character of our study, the results provide a rationale for further clarifying the role of the identified miRNAs in the pathogenesis of PD and for validating their diagnostic potential.
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http://dx.doi.org/10.1212/WNL.0000000000001258DOI Listing
February 2015

Identification of a set of endogenous reference genes for miRNA expression studies in Parkinson's disease blood samples.

BMC Res Notes 2014 Oct 10;7:715. Epub 2014 Oct 10.

Center for Biomedicine, European Academy Bozen/Bolzano (EURAC), 39100 Bolzano, Italy, Affiliated Institute of the University of Lübeck, Lübeck, Germany.

Background: Research on microRNAs (miRNAs) is becoming an increasingly attractive field, as these small RNA molecules are involved in several physiological functions and diseases. To date, only few studies have assessed the expression of blood miRNAs related to Parkinson's disease (PD) using microarray and quantitative real-time PCR (qRT-PCR). Measuring miRNA expression involves normalization of qRT-PCR data using endogenous reference genes for calibration, but their choice remains a delicate problem with serious impact on the resulting expression levels. The aim of the present study was to evaluate the suitability of a set of commonly used small RNAs as normalizers and to identify which of these miRNAs might be considered reliable reference genes in qRT-PCR expression analyses on PD blood samples.

Results: Commonly used reference genes snoRNA RNU24, snRNA RNU6B, snoRNA Z30 and miR-103a-3p were selected from the literature. We then analyzed the effect of using these genes as reference, alone or in any possible combination, on the measured expression levels of the target genes miR-30b-5p and miR-29a-3p, which have been previously reported to be deregulated in PD blood samples.

Conclusions: We identified RNU24 and Z30 as a reliable and stable pair of reference genes in PD blood samples.
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http://dx.doi.org/10.1186/1756-0500-7-715DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4209045PMC
October 2014

The mitochondrial calcium uniporter is a multimer that can include a dominant-negative pore-forming subunit.

EMBO J 2013 Aug 30;32(17):2362-76. Epub 2013 Jul 30.

Department of Biomedical Sciences, University of Padua and CNR Neuroscience Institute, Padua, Italy.

Mitochondrial calcium uniporter (MCU) channel is responsible for Ruthenium Red-sensitive mitochondrial calcium uptake. Here, we demonstrate MCU oligomerization by immunoprecipitation and Förster resonance energy transfer (FRET) and characterize a novel protein (MCUb) with two predicted transmembrane domains, 50% sequence similarity and a different expression profile from MCU. Based on computational modelling, MCUb includes critical amino-acid substitutions in the pore region and indeed MCUb does not form a calcium-permeable channel in planar lipid bilayers. In HeLa cells, MCUb is inserted into the oligomer and exerts a dominant-negative effect, reducing the [Ca(2+)]mt increases evoked by agonist stimulation. Accordingly, in vitro co-expression of MCUb with MCU drastically reduces the probability of observing channel activity in planar lipid bilayer experiments. These data unveil the structural complexity of MCU and demonstrate a novel regulatory mechanism, based on the inclusion of dominant-negative subunits in a multimeric channel, that underlies the fine control of the physiologically and pathologically relevant process of mitochondrial calcium homeostasis.
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http://dx.doi.org/10.1038/emboj.2013.157DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3771344PMC
August 2013

Reg3G gene expression in regenerating skeletal muscle and corresponding nerve.

Muscle Nerve 2014 Jan 5;49(1):61-8. Epub 2013 Dec 5.

Department of Anatomy, School of Medicine, University of Rijeka, Brace Branchetta 20, 51000, Rijeka, Croatia.

Introduction: The Reg genes play a major role in the regeneration of various tissues; however, no reports have been published regarding expression of the Reg3G gene in skeletal muscle. In this study we investigated the expression of the Reg3G gene in regeneration of rat skeletal muscle and injured nerves.

Methods: We used 3 experimental models of muscle and nerve injury. RT-PCR and Western blot analysis were performed for detection of Reg3G in regenerating muscle and nerve.

Results: We found transcriptional activation of the Reg3G gene in the soleus and extensor digitorum longus muscles and in their corresponding nerves after both muscle and nerve injury in different time periods, respectively.

Conclusions: The results suggest that the Reg3G gene plays a major role in communication between injured axons and muscle and may play a significant role in skeletal muscle and peripheral nerve regeneration.
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http://dx.doi.org/10.1002/mus.23877DOI Listing
January 2014

Adaptation of mouse skeletal muscle to long-term microgravity in the MDS mission.

PLoS One 2012 28;7(3):e33232. Epub 2012 Mar 28.

Department of Biomedical Sciences, University of Padova, Padova, Italy.

The effect of microgravity on skeletal muscles has so far been examined in rat and mice only after short-term (5-20 day) spaceflights. The mice drawer system (MDS) program, sponsored by Italian Space Agency, for the first time aimed to investigate the consequences of long-term (91 days) exposure to microgravity in mice within the International Space Station. Muscle atrophy was present indistinctly in all fiber types of the slow-twitch soleus muscle, but was only slightly greater than that observed after 20 days of spaceflight. Myosin heavy chain analysis indicated a concomitant slow-to-fast transition of soleus. In addition, spaceflight induced translocation of sarcolemmal nitric oxide synthase-1 (NOS1) into the cytosol in soleus but not in the fast-twitch extensor digitorum longus (EDL) muscle. Most of the sarcolemmal ion channel subunits were up-regulated, more in soleus than EDL, whereas Ca(2+)-activated K(+) channels were down-regulated, consistent with the phenotype transition. Gene expression of the atrophy-related ubiquitin-ligases was up-regulated in both spaceflown soleus and EDL muscles, whereas autophagy genes were in the control range. Muscle-specific IGF-1 and interleukin-6 were down-regulated in soleus but up-regulated in EDL. Also, various stress-related genes were up-regulated in spaceflown EDL, not in soleus. Altogether, these results suggest that EDL muscle may resist to microgravity-induced atrophy by activating compensatory and protective pathways. Our study shows the extended sensitivity of antigravity soleus muscle after prolonged exposition to microgravity, suggests possible mechanisms accounting for the resistance of EDL, and individuates some molecular targets for the development of countermeasures.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0033232PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3314659PMC
August 2012

Reg IV protein and mRNA expression in different rat organs.

Acta Histochem 2011 Dec 17;113(8):793-7. Epub 2010 Dec 17.

Department of Anesthesiology and Intensive Care, University Hospital Rijeka, Kresimirova 42, Rijeka, Croatia.

The Reg IV gene has been documented in the human colon, small intestine, stomach and pancreas. Expression of the Reg IV in different cell types has been associated with regeneration, cell growth and cell survival, cell adhesion and resistance to apoptosis. Since the distribution of the Reg IV protein in normal rat tissues is unknown, the aim of this study was to reveal the expression of the Reg IV protein in structurally and functionally different rat organs. The expression of Reg IV gene was analyzed by Western blot and reverse transcription-polymerase chain reaction. Immunohistochemistry was used to localize Reg IV protein. Reg IV protein was expressed in pancreas, stomach, small intestine, colon, brain, spleen, kidney and urinary bladder in two-month-old male Wistar rats. In addition, the expression of Reg IV mRNA by reverse transcription-polymerase chain reaction was confirmed. Our study provides detailed information about the expression and localization of Reg IV protein in different rat organs. These findings provide an evidence of Reg IV expression in different rat organs, which may help elucidate a potential role in growth and proliferation of different cells like other members of the Reg family genes which act as growth factors in the different organs.
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http://dx.doi.org/10.1016/j.acthis.2010.11.008DOI Listing
December 2011

Long-term recurrence of secretory breast carcinoma with metastatic sentinel lymph nodes.

Arch Gynecol Obstet 2011 Mar 14;283 Suppl 1:77-8. Epub 2010 Sep 14.

Department of Oncological Surgery, Georges François Leclerc Anticancer Centre, 1, rue du Professeur Marion, 21000, Dijon, France.

Purpose: Secretory carcinoma is a rare form of breast cancer most frequently encountered in children or young adults. Several cases have been described in adults with increased aggressiveness and a risk of metastases.

Case Report: We report here, in a 30-year-old woman, a case of local relapse and lymph node metastases occurring 16 years after the initial diagnosis of secretory carcinoma.

Conclusion: We present the clinical, radiological and pathological findings that led to the diagnosis.
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http://dx.doi.org/10.1007/s00404-010-1669-9DOI Listing
March 2011

Three-dimensional sonographic diagnosis of abdominal wall endometriosis: a useful tool?

Fertil Steril 2011 Jan 20;95(1):289.e1-4. Epub 2010 Jun 20.

Department of Obstetric, Gynecology, and Reproductive Medicine, University of Saint Etienne, Jean Monnet, Saint-Etienne, France.

Objective: To report the usefulness of three-dimensional (3D) ultrasonography for the assessment of parietal endometriosis.

Design: Case report.

Setting: Academic research hospital.

Patient(s): A 35-year-old woman with a noncyclic, painful abdominal nodule near a caesarean delivery scar.

Intervention(s): 3D ultrasonography and wide surgical resection with healthy margins.

Main Outcome Measure(s): 3D ultrasonographic assessment of the endometriotic nodule.

Result(s): We found that 3D ultrasonography offered a more specific description of parietal endometriosis with irregular and spiculated margins and depth infiltration as well as provided preoperative evaluation of volume measurements.

Conclusion(s): Three-dimensional ultrasonography is a useful, noninvasive tool in extrapelvic endometriosis.
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http://dx.doi.org/10.1016/j.fertnstert.2010.05.027DOI Listing
January 2011

NFAT isoforms control activity-dependent muscle fiber type specification.

Proc Natl Acad Sci U S A 2009 Aug 24;106(32):13335-40. Epub 2009 Jul 24.

Department of Biomedical Sciences, University of Padova, 35131 Padova, Italy.

The intracellular signals that convert fast and slow motor neuron activity into muscle fiber type specific transcriptional programs have only been partially defined. The calcium/calmodulin-dependent phosphatase calcineurin (Cn) has been shown to mediate the transcriptional effects of motor neuron activity, but precisely how 4 distinct muscle fiber types are composed and maintained in response to activity is largely unknown. Here, we show that 4 nuclear factor of activated T cell (NFAT) family members act coordinately downstream of Cn in the specification of muscle fiber types. We analyzed the role of NFAT family members in vivo by transient transfection in skeletal muscle using a loss-of-function approach by RNAi. Our results show that, depending on the applied activity pattern, different combinations of NFAT family members translocate to the nucleus contributing to the transcription of fiber type specific genes. We provide evidence that the transcription of slow and fast myosin heavy chain (MyHC) genes uses different combinations of NFAT family members, ranging from MyHC-slow, which uses all 4 NFAT isoforms, to MyHC-2B, which only uses NFATc4. Our data contribute to the elucidation of the mechanisms whereby activity can modulate the phenotype and performance of skeletal muscle.
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http://dx.doi.org/10.1073/pnas.0812911106DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2726382PMC
August 2009

Comparative transcriptional and biochemical studies in muscle of myotonic dystrophies (DM1 and DM2).

Neurol Sci 2009 Jun 27;30(3):185-92. Epub 2009 Mar 27.

Department of Biomedical Sciences, University of Padova, viale G. Colombo 3, Padua, Italy.

Myotonic dystrophy type 1 (DM1) and myotonic dystrophy type 2 (proximal muscular myopaty/DM2) are caused by similar dynamic mutations at two distinct genetic loci. The two diseases also lead to similar phenotypes but different clinical severity. Dysregulation of alternative splicing has been suggested as the common pathogenic mechanism. Here, we investigate the molecular differences between DM1 and DM2 using reverse transcriptase-polymerase chain reaction of troponin T (TnT) and the insulin receptor (IR), as well as immunoblotting of TnT in muscle biopsies from DM1 and DM2 patients. We found that: (a) slow TnT was encoded by two different transcripts in significantly different ratios in DM1 and DM2 muscles; (b) DM2 muscles exhibited a higher degree of alternative splicing dysregulation for fast TnT transcripts when compared to DM1 muscles; (c) the distribution of TnT proteins was significantly skewed towards higher molecular weight species in both diseases; (d) the RNA for the insulin-independent IR-A isoform was significantly increased and appeared related to the fibre-type composition in the majority of the cases examined. On the whole, these data should give a better insight on pathogenesis of DM1 and DM2.
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http://dx.doi.org/10.1007/s10072-009-0048-4DOI Listing
June 2009

Reg IV protein is expressed in normal rat tissue.

Coll Antropol 2008 Oct;32 Suppl 2:89-93

Department of Anatomy, School of Medicine, University of Rijeka, Rijeka, Croatia.

The Reg IV gene has been documented in the colon, small intestine, stomach and pancreas of the human. Expression of the Reg IV in different cell types has been associated with regeneration, cell growth and cell survival, cell adhesion and resistance to apoptosis. It is unknown whether the Reg IV protein is present in the normal rat tissue. The aim of this study was to reveal the expression of the Reg IV protein in the rat spleen and colon. Western blot analysis using antibody specific for Reg IV protein were performed on rat spleen and colon extracts. Low level of Reg IV expression was found in all examined colon samples. The expression of Reg IV protein in spleen tissue was significantly higher than in the colon. Reg IV protein was immunohistochemically stained in a few epithelial cells in the basal portion of colon crypts and in a large spleen cells which were scattered in the red pulp. Our results demonstrate for the first time the presence of the Reg IV protein expression in the healthy spleen and colon tissue of the rat. Other members of the Reg family, Reg I and Reg III proteins have been shown to act as a growth factors in gastrointestinal tract, but without further experiments we can only assume the potential role of the Reg IV protein in spleen and colon cell growth.
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October 2008

Tetraarylphosphonium-supported carbodiimide reagents: synthesis, structure optimization and applications.

J Org Chem 2008 Apr 4;73(7):2542-7. Epub 2008 Mar 4.

Département de Chimie, Université de Montréal, PO Box 6128, Station Downtown, Montréal, Canada QC H3C 3J7.

New tetraarylphosphonium (TAP)-supported alkyl- and arylcarbodiimides were synthesized and used as coupling reagents for esterification reactions, amidation reactions and dehydration reactions of hydroxyesters. Taking advantage of the solubility properties imparted by the tetraarylphosphonium unit, a simple precipitation and filtration allowed complete separation of the urea by-products. This paper describes the structure optimization study of the various TAP-supported carbodiimide reagents to obtain the desired reactivity and solubility profile. Furthermore, we have demonstrated that the diimide reagent can be regenerated from the urea to recycle the reagents.
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http://dx.doi.org/10.1021/jo702417vDOI Listing
April 2008

GATA elements control repression of cardiac troponin I promoter activity in skeletal muscle cells.

BMC Mol Biol 2007 Sep 17;8:78. Epub 2007 Sep 17.

Department of Biomedical Sciences, University of Padova, Padova, Italy.

Background: We reported previously that the cardiac troponin I (cTnI) promoter drives cardiac-specific expression of reporter genes in cardiac muscle cells and in transgenic mice, and that disruption of GATA elements inactivates the cTnI promoter in cultured cardiomyocytes. We have now examined the role of cTnI promoter GATA elements in skeletal muscle cells.

Results: Mutation or deletion of GATA elements induces a strong transcriptional activation of the cTnI promoter in regenerating skeletal muscle and in cultured skeletal muscle cells. Electrophoretic mobility shift assays show that proteins present in nuclear extracts of C2C12 muscle cells bind the GATA motifs present in the cTnI promoter. However, GATA protein complex formation is neither reduced nor supershifted by antibodies specific for GATA-2, -3 and -4, the only GATA transcripts present in muscle cells.

Conclusion: These findings indicate that the cTnI gene promoter is repressed in skeletal muscle cells by GATA-like factors and open the way to further studies aimed at identifying these factors.
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http://dx.doi.org/10.1186/1471-2199-8-78DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2045674PMC
September 2007

Foxo transcription factors induce the atrophy-related ubiquitin ligase atrogin-1 and cause skeletal muscle atrophy.

Cell 2004 Apr;117(3):399-412

Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.

Skeletal muscle atrophy is a debilitating response to fasting, disuse, cancer, and other systemic diseases. In atrophying muscles, the ubiquitin ligase, atrogin-1 (MAFbx), is dramatically induced, and this response is necessary for rapid atrophy. Here, we show that in cultured myotubes undergoing atrophy, the activity of the PI3K/AKT pathway decreases, leading to activation of Foxo transcription factors and atrogin-1 induction. IGF-1 treatment or AKT overexpression inhibits Foxo and atrogin-1 expression. Moreover, constitutively active Foxo3 acts on the atrogin-1 promoter to cause atrogin-1 transcription and dramatic atrophy of myotubes and muscle fibers. When Foxo activation is blocked by a dominant-negative construct in myotubes or by RNAi in mouse muscles in vivo, atrogin-1 induction during starvation and atrophy of myotubes induced by glucocorticoids are prevented. Thus, forkhead factor(s) play a critical role in the development of muscle atrophy, and inhibition of Foxo factors is an attractive approach to combat muscle wasting.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3619734PMC
http://dx.doi.org/10.1016/s0092-8674(04)00400-3DOI Listing
April 2004

Heart morphogenesis is not affected by overexpression of the Sh3bgr gene mapping to the Down syndrome heart critical region.

Hum Genet 2004 Apr 7;114(5):517-9. Epub 2004 Feb 7.

Venetian Institute of Molecular Medicine, Via Orus 2, 35129 Padua, Italy.

Congenital heart disease (CHD) is the most common birth defect in humans and is present in 40% of newborns affected by Down syndrome (DS). The SH3BGR gene maps to the DS-CHD region and is a potential candidate for the pathogenesis of CHD, since it is selectively expressed in cardiac and skeletal muscle. To determine whether overexpression of Sh3bgr in the murine heart may cause abnormal cardiac development, we have generated transgenic mice using a cardiac- and skeletal-muscle-specific promoter to drive the expression of a Sh3bgr transgene. We report here that heart morphogenesis is not affected by overexpression of Sh3bgr.
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http://dx.doi.org/10.1007/s00439-004-1088-8DOI Listing
April 2004

Highly substituted pyrrolidinones and pyridones by 4-CR/2-CR sequence.

Org Lett 2004 Jan;6(1):39-42

Morphochem AG, Gmunderstr. 37-37a, 81379 Munich, Germany.

[reaction: see text] By combining a Ugi four-component reaction of isocyanides, phosphonoacetic acids, primary amines, and glyoxals or alternatively 3-keto aldehydes with a subsequent Wittig ring-closing reaction (using the Horner/Wadsworth/Emmons variant (HWE)), highly substituted 5-oxo-2,5-dihydro-1H-pyrrole-2-carboxylic acid amides and 6-oxo-1,2,3,6-tetrahydro-pyridine-2-carboxylic acid amides can be assembled, respectively. The corresponding tandem of a Passerini reaction on 3-keto aldehydes and subsequent Wittig ring closure does not afford the expected six-membered 6-oxo-3,6-dihydro-2H-pyran-2-carboxylic acid amides but instead leads to the formation of 4-oxo-pent-2-enoic acid amides via an elimination route.
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http://dx.doi.org/10.1021/ol035787nDOI Listing
January 2004

Short and diverse route toward complex natural product-like macrocycles.

Org Lett 2003 Apr;5(7):1047-50

Morphochem AG, Gmunderstr. 37-37a, 81379 München, Germany.

[reaction: see text] A general strategy toward macrocyclic compounds using multicomponent reaction (MCR) chemistry, e.g., Passerini and Ugi variants, and ring-closing metathesis (RCM) is introduced. The corresponding bifunctional isocyanides carboxylic acids bearing a terminal olefin are easy to prepare from the corresponding commercially available starting materials. Advantageously, this strategy allows fast access to a diverse conformational space of natural product-like macrocycles and could thus be of interest in the discovery of novel bioactive agents.
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http://dx.doi.org/10.1021/ol034077eDOI Listing
April 2003