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Up-regulated microRNAs in blastocoel fluid of human implanted embryos could control circuits of pluripotency and be related to embryo competence

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JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 2025年 1-15页

作者： Battaglia, Rosalia Caponnetto, Angela Ferrara, Carmen Fazzio, Anna Barbagallo, Cristina Stella, Michele Barbagallo, Davide Ragusa, Marco Vento, Maria Elena Borzi, Placido Scollo, Paolo carli, luca Feichtinger, Michael Kasapi, Evangelia Tsakos, Elias Palini, Simone Sierka, Wojciech Pecorino, Basilio Campitiello, Maria Rosaria Ronsini, Carlo Purrello, Michele Valerio, Domenico Longobardi, Salvatore D'Hooghe, Thomas Di Pietro, Cinzia Univ Catania Dept Biomed & Biotechnol Sci Sect Biol & Genet G Sichel I-95123 Catania Italy Univ Enna Kore Dept Med & Surg I-94100 Enna Italy Cannizzaro Hosp IVF Unit Catania Italy Wunschbaby Inst Feichtinger Vienna Austria EmbryoClin IVF Thessaloniki Greece Cervesi Hosp IVF Unit Cattolica Italy Gyncentrum Sp Zelazna 1 PL-40851 Katowice Poland ASL Salerno Dept Obstet & Gynecol & Physiopathol Human Reprod Salerno Italy Univ Campania Luigi Vanvitelli Dept Woman Child & Gen & Specialized Surg Naples Italy Inst Genet Res IRG I-80143 Naples Italy Merck Serono SpA Global Clin Dev Rome Italy Merck KGaA Darmstadt Germany KU Leuven Univ Leuven Dept Dev & Regenerat Biomed Sci Grp Leuven Belgium

Purpose The paper aims to investigate the biological role of microRNAs secreted by preimplantation embryo into the blastocoel fluid and to detect a distinctive molecular signature for identifying embryos with the highest implantation potential. Methods We carried on a multicenter retrospective study involving five European IVF centers. We collected 112 blastocoel fluid samples from embryos on day 5 post-fertilization, cultured individually, along with data on blastocyst grade and embryo transfer outcomes. Using a custom TLDA Array, we compared the expression levels of 89 miRNAs between 33 fluids from high-quality implanted embryos and 30 fluids from high-quality not-implanted embryos. Expression differences were assessed using SAM and t-test. Additionally, correlation and function enrichment analysis and network construction were conducted to identify the biological roles of deregulated microRNAs. Results We identified six up-regulated microRNAs in the blastocoel fluid from implanted embryos, significantly and positively correlated across all samples (r >= 0.7;P <= 0.05). They could take part in pluripotency circuits, regulating and being regulated by transcription factors associated with stemness, cell growth, and embryo development. The ROC curve analysis confirmed the potential of these miRNAs as implantation classifiers. Conclusion The six miRNAs up-regulated in blastocoel fluid from implanted embryos may represent a functional molecular signature for evaluating blastocyst quality and identifying the most competent embryos. Their evaluation associated with non-invasive preimplantation genetic testing, integrating epigenetic and genomic analyses, could enhance implantation grade and allow for identification of the euploid embryo not able to implant.

关键词： Blastocoel fluid Embryo development Embryo implantation Human blastocyst MicroRNAs

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ASSAY OF DIFFUSION OF DIFFERENT BOTULINUM NEUROTOXIN TYPE A FORMULATIONS INJECTED IN THE MOUSE LEG

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MUSCLE & NERVE 2009年第3期40卷 374-380页

作者： carli, luca Montecucco, Cesare Rossetto, Ornella Univ Padua Dept Biomed Sci Padua Italy

Botulinum neurotoxin type-A (BoNT/A) is very effective in the therapy of a wide range of human syndromes characterized by hyperactivity of peripheral cholinergic nerve terminals. Little diffusion of this toxin from the site of injection is commonly observed, but even minor changes in this property would greatly affect the validity of the treatment. Different pharmacological formulations of BoNT/A are available, and they may have different diffusion characteristics due to protein complex size, product format, and pharmacological properties. Here we assessed the extent of diffusion of three commercial preparations of BoNT/A: Botox (Allergan), Dysport (Ipsen), and Xeomin (Merz Pharmaceuticals) using a novel and highly sensitive test based on neural cell adhesion molecule (N-CAM) expression in muscle. N-CAM is a membrane glycoprotein that accumulates on muscle fibers after denervation and is not expressed in untreated adult muscle. This allows fine monitoring of the functional diffusion of this toxin, and the sensitivity of this assay is emphasized by the use of the mouse model because of the small muscle dimensions. The results presented here indicate that there is no significant difference between Botox, Dysport, and Xeomin with respect to diffusion into adjacent muscles in the mouse leg. Muscle Nerve 40: 374-380, 2009

关键词： botulinum toxin type A neuromuscular junction N-CAM diffusion

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Random spray Retinex: A new Retinex implementation to investigate the local properties of the model

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IEEE TRANSACTIONS ON IMAGE PROCESSING 2007年第1期16卷 162-171页

作者： Provenzi, Edoardo Fierro, Massimo Rizzi, Alessandro De carli, luca Gadia, Davide Marini, Daniele Univ Milan Dipartimento Tecnol Informaz I-26013 Crema Italy Univ Milan Dipartimento Informat & Comunicaz I-20135 Milan Italy

In order to investigate the local filtering behavior of the Retinex model, we propose a new implementation in which paths are replaced by 2-D pixel sprays, hence the name "random spray Retinex." A peculiar feature of this implementation is the way its parameters can be controlled to perform spatial investigation. The parameters' tuning is accomplished by an unsupervised method based on quantitative measures. This procedure has been validated via user panel tests. Furthermore, the spray approach has faster performances than the path-wise one. Tests and results are presented and discussed.

关键词： locality of color perception pixel sprays Retinex

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Benefits of Selecting PMASR Machines as Traction Motors in Battery Electric Forklift Trucks

Benefits of Selecting PMASR Machines as Traction Motors in B...

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IEEE International Conference on Electrical Systems for Aircraft, Railway, Ship Propulsion and Road Vehicles / International Transportation Electrification Conference (ESARS-ITEC)

作者： Olivo, Matteo Tessarolo, Alberto De carli, luca Pozzo, Riccardo Univ Trieste Dept Engn & Architecture Trieste Italy PMP Ind Grp Elect Vehicle Driveline Div Coseano UD Italy

ISBN: (纸本)9798350346893

The advantages of using PM-assisted synchronous reluctance motors as traction devices for electric forklift vehicles are investigated in this paper. Motor energy consumption and drive overall performance are analytically estimated through a simulation of a standard VDI-based driving cycle and compared with the corresponding results from a reference system driven using induction motors.

关键词： electric vehicles traction motors energy efficiency brushless machines industry applications

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Tetanus, Botulinum and Snake Presynaptic Neurotoxins

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RENDICONTI LINCEI-SCIENZE FISICHE E NATURALI 2008年第2期19卷 173-188页

作者： Rossetto, Ornella Morbiato, Laura Caccin, Paola Rigoni, Michela carli, luca Paoli, Marco Cintra-Francischelli, Mariana Montecucco, Cesare Univ Padua Dipartimento Sci Biomed I-35121 Padua Italy Univ Padua Ist CNR Neurosci I-35121 Padua Italy

Tetanus and botulinum neurotoxins, produced by anaerobic bacteria of the genus Clostridium, are the most toxic proteins known and are solely responsible for the pathogenesis of tetanus and botulism. They are metalloproteases that enter nerve terminals and cleave proteins of the neuroexocytosis apparatus causing a persistent, but reversible, inhibition of neurotransmitter release. Botulinum neurotoxins are used in the therapy of many human syndromes caused by hyperactive nerve terminals. Snake presynaptic PLA2 neurotoxins block nerve terminals by binding to the nervemembrane and catalyzing phospholipid hydrolysis with production of lysophospholipids and fatty acids. These compounds change the membrane conformation causing enhanced fusion of synaptic vesicle via hemifusion intermediate with release of neurotransmitter and, at the same time, inhibition of vesicle fission and recycling. It is possible to envisage clinical applications of the lysophospholipid/fatty acid mixture to inhibit hyperactive superficial nerve terminals.

关键词： Tetanus and botulinum neurotoxins snake neurotoxins neuroexocytosis muscle paralysis dystonia

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Non-invasive preimplantation genetic screening using array comparative genomic hybridization on spent culture media: a proof-of-concept pilot study

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REPRODUCTIVE BIOMEDICINE ONLINE 2017年第6期34卷 583-589页

作者： Feichtinger, Michael Vaccari, Enrico carli, luca Wallner, Elisabeth Maedel, Ulrike Figl, Katharina Palini, Simone Feichtinger, Wilfried Wunschbaby Inst Feichtinger Lainzerstr 6 A-1130 Vienna Austria Med Univ Vienna Div Gynecol Endocrinol & Reprod Med Dept Obstet & Gynaecol Waehringer Guertel 18-20 A-1090 Vienna Austria Cervesi Hosp Cattolica IVF Unit I-47841 Cattolica Rimini Italy

The aim of this pilot study was to assess if array comparative genomic hybridization (aCGH) non-invasive preimplantation genetic screening (PGS) on blastocyst culture media is feasible. Therefore, aCGH analysis was carried out on 22 spent blastocyst culture media samples after polar body PGS because of advanced maternal age. All oocytes were fertilized by intracytoplasmic sperm injection and all embryos underwent assisted hatching. Concordance of polar body analysis and culture media genetic results was assessed. Thirteen out of 18 samples (72.2%) revealed general concordance of ploidy status (euploid or aneuploid). At least one chromosomal aberration was found concordant in 10 out of 15 embryos found to be aneuploid by both polar body and culture media analysis. Overall, 17 out of 35 (48.6%) single chromosomal aneuploidies were concordant between the culture media and polar body analysis. By analysing negative controls (oocytes with fertilization failure], notable maternal contamination was observed. Therefore, non-invasive PGS could serve as a second matrix after polar body or cleavage stage PGS;however, in euploid results, maternal contamination needs to be considered and results interpreted with caution. (C) 2017 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

关键词： NIPGS PGS Non-invasive PGD Culture-medium

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Evaluation of Nanopore Sequencing on Polar Bodies for Routine Pre-Implantation Genetic Testing for Aneuploidy

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CLINICAL CHEMISTRY 2024年第5期70卷 747-758页

作者： Oberle, Anna Hanzer, Franziska Kokocinski, Felix Ennemoser, Anna carli, luca Vaccari, Enrico Hengstschlaeger, Markus Feichtinger, Michael Wunschbaby Inst Feichtinger Lainzer Str 6 A-1130 Vienna Austria Gene Test Bioinformat Solut GmbH Jakob Muller Str 16 D-68623 Lampertheim Germany

Background Preimplantation genetic testing for aneuploidy (PGT-A) using polar body (PB) biopsy offers a clinical benefit by reducing the number of embryo transfers and miscarriage rates but is currently not cost-efficient. Nanopore sequencing technology opens possibilities by providing cost-efficient and fast sequencing results with uncomplicated sample preparation work *** In this comparative experimental study, 102 pooled PB samples (99 passing QC) from 20 patients were analyzed for aneuploidy using nanopore sequencing technology and compared with array comparative genomic hybridization (aCGH) results generated as part of the clinical routine. Samples were sequenced on a Nanopore MinION machine. Whole-chromosome copy-numbers were called by custom bioinformatic analysis software. Automatically called results were compared to aCGH *** Overall, 96/99 samples were consistently detected as euploid or aneuploid in both methods (concordance = 97.0%, sensitivity = 0.957, specificity = 1.0, positive predictive value = 1.0, negative predictive value = 0.906). On the chromosomal level, concordance reached 98.7%. Chromosomal aneuploidies analyzed in this trial covered all 23 chromosomes with 98 trisomies, and 97 monosomies in 70 aCGH samples. The whole nanopore work flow is feasible in under 5 h (for one sample) with a maximum time of 16 h (for 12 samples), enabling fresh PB-euploid embryo transfer. A material cost of US$ 165 (EUR 150)/sample possibly enables cost-efficient aneuploidy *** Overall, 96/99 samples were consistently detected as euploid or aneuploid in both methods (concordance = 97.0%, sensitivity = 0.957, specificity = 1.0, positive predictive value = 1.0, negative predictive value = 0.906). On the chromosomal level, concordance reached 98.7%. Chromosomal aneuploidies analyzed in this trial covered all 23 chromosomes with 98 trisomies, and 97 monosomies in 70 aCGH samples. The whole nanopore work flow is feasible in under 5 h

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Different mechanisms of inhibition of nerve terminals by botulinum and snake presynaptic neurotoxins

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TOXICON 2009年第5期54卷 561-564页

作者： Montecucco, Cesare Rossetto, Ornella Caccin, Paola Rigoni, Michela carli, luca Morbiato, Laura Muraro, Lucia Paoli, Marco Univ Padua Dipartimento Sci Biomed I-35131 Padua Italy Univ Padua Ist CNR Neurosci I-35131 Padua Italy

The different mode of action on peripheral nerve terminals of the botulinum neurotoxins and of the snake presynaptic phospholipase A2 neurotoxins is reviewed here. These two groups of toxins are highly toxic because they are neurospecific and at the same time are enzymes that can modify many substrate molecules before being inactivated. The similarity of symptoms they cause in humans derives from the fact that both botulinum neurotoxins (seven serotypes named A-G) and snake presynaptic PLA2 neurotoxins block the nerve terminals and that peripheral cholinergic terminals are major targets. Given this general similarity of targets and clinical symptoms, the specific molecular and cellular mechanisms at the basis of their action are very different. This difference appears evident from the beginning of intoxication, i.e. neurotoxins binding to peripheral nerve terminals and proceeds with the different site of actions and molecular targets. (C) 2008 Elsevier Ltd. All rights reserved.

关键词： Botulinum neurotoxins Snake presynaptic PLA2 neurotoxins

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Improve adherence to the Mediterranean diet through an innovative app: a pilot study

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NUTRITION & FOOD SCIENCE 2023年第1期53卷 138-147页

作者： Devecchi, Andrea Bo, Simona De carli, luca Breda, Erik Ponzo, Valentina Pezzana, Andrea Univ Gastron Sci Pollenzo Bra Italy Univ Torino Dept Med Sci Turin Italy ASL Citta Torino SC Clin Nutr Turin Italy

Purpose The Mediterranean diet (MD) is a sustainable and healthy diet. However, compliance to the MD is still poor. Given this, the authors created a Web app to promote the MD. The purpose of this study was to evaluate the efficacy of the Web app YourPappa in terms of adherence to the MD. Design/methodology/approach The authors developed a Web app, YourPappa, with the aim of encouraging virtuous dietary habits through a reward mechanism. After that, a randomized controlled study was conducted. All participants were given written advice on correct nutrition. Moreover, the case group was trained on the use of the app. The MD adherence was evaluated by a validated questionnaire (Medi-Lite). Findings Cases found an average increase in the Medi-Lite scores of +7.1%, whereas controls showed an increase of +0.7% (p = 0.06;effect size 0.60). For most of the users, the Web app helped them to think about what they were buying and to promote the MD. Research limitations/implications Obesity and related diseases are a topical problem. New strategies are needed to counter it. This study showed interesting and encouraging results, which need further research and insight to be validated and supported. Originality/value To the best of the authors' knowledge, this is the first study that has evaluated the use of a Web app to stimulate the adoption of the MD through a reward mechanism.

关键词： Mediterranean diet Health Technology Innovation Web application Reward Nutrition Food

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Longitudinal study of quality of life in advanced cancer patients on home parenteral nutrition

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CANCER MEDICINE 2017年第7期6卷 1799-1806页

作者： Cotogni, Paolo De carli, luca Passera, Roberto Amerio, Maria Luisa Agnello, Elena Fadda, Maurizio Ossola, Marta Monge, Taira De Francesco, Antonella Bozzetti, Federico Univ Turin Dept Anesthesia & Intens Care Turin Italy S Giovanni Battista Hosp Unit Parenteral Nutr Oncol Turin Italy S Giovanni Battista Hosp Clin Nutr Turin Italy S Giovanni Battista Hosp Nucl Med Div Turin Italy Cardinal Massaia Hosp Clin Nutr Asti Italy Univ Milan Fac Med Milan Italy

Since there is little knowledge regarding the quality of life (QoL) of cancer patients on home parenteral nutrition (HPN), we planned a prospective, longitudinal, double-center study to investigate the changes of QoL in these patients. One hundred and eleven adult cancer patients who were candidates for HPN following the indications of the European guidelines were consecutively enrolled. For QoL analysis, EORTC QLQ-C30 questionnaires were filled at the HPN start and after 1, 2, 3, and 4months, and scores changes over time were analyzed according to the univariate mixed-effects linear model for repeated measures. Most patients had gastrointestinal cancers, were severely malnourished, and were in stage IV;two-thirds were still receiving oncologic treatments. Median weight loss over 3months and body mass index were 11.7% and 20.7, respectively. Median survival was 4.7 (1-42) months;67 and 34% of patients survived 3 and 6months, respectively. Global QoL, physical functioning, role functioning, emotional functioning, appetite loss, and fatigue scores had a statistically significant trend over time (P<0.001, P<0.001, P=0.007, P<0.001, P=0.004, P=0.022, respectively). At the univariate analyses, the determinants significantly associated with changes in trend over time for physical, role, and emotional functioning were oncologic treatments (P<0.001, P=0.014, P=0.040, respectively) and for appetite loss they were weight loss and Karnofsky performance status (P=0.003, P=0.023, respectively). Global QoL, physical, role, and emotional functioning improved during HPN even in advanced cancer patients on oncologic treatments.

关键词： EORTC QLQ-C30 questionnaire home care nutrition oncologic treatment palliative care quality of life supportive care

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