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A LIM Domain Protein from Tobacco Involved in Actin-Bundling and Histone Gene Transcription

Molecular Plant 2013年第2期6卷 483-502页

作者： Danièle Moes sabrina gatti CEline Hoffmann Monika Dieterle Flora Moreau Katrin Neumann Marc Schumacher Marc Diederich Erwin Grill Wen-Hui Shen André Steinmetz Cldment Thomas Centre de Recherche Public-SantE 84 Val Fleuri L-1526 Luxembourg Luxembourg Laboratoire de Biologie Moleculaire et Cellulaire du Cancer HSpital Kirchberg 9 rue Edward Steichen L-2540 Luxembourg Luxembourg Lehrstuhl for Botanik Technische Universitat Munchen EmiI-Ramann-StraBe 4 D-85354 Freising Germany Jnstitut de Biologie Moleculaire des Plantes du CNRS 12 rue du Général Zimmer F-67084 Strasbourg Ceédex France

The two LIM domain-containing proteins from plants （LIMs） typically exhibit a dual cytoplasmic-nuclear dis-tribution, suggesting that, in addition to their previously described roles in actin cytoskeleton organization, they partici-pate in nuclear processes. Using a south-western blot-based screen aimed at identifying factors that bind to plant histone gene promoters, we isolated a positive clone containing the tobacco LIM protein WLIM2 （NtWLIM2） cDNA. Using both green fluorescent protein （GFP） fusion-and immunology-based strategies, we provide clear evidence that NtWLIM2 local-izes to the actin cytoskeleton, the nucleus, and the nucleolus. Interestingly, the disruption of the actin cytoskeleton by latrunculin B significantly increases NtWLIM2 nuclear fraction, pinpointing a possible novel cytoskeletal-nuclear crosstalk. Biochemical and electron microscopy experiments reveal the ability of NtWLIM2 to directly bind to actin filaments and to crosslink the latter into thick actin bundles. Electrophoretic mobility shift assays show that NtWLIM2 specifically binds to the conserved octameric cis-elements （Oct） of the Arabidopsis histone H4A748 gene promoter and that this binding largely relies on both LIM domains. Importantly, reporter-based experiments conducted in Arabidopsis and tobacco proto-plasts confirm the ability of NtWLIM2 to bind to and activate the H4A748 gene promoter in live cells. Expression studies indicate the constitutive presence of NtWLIM2 mRNA and NtWLIM2 protein during tobacco BY-2 cell proliferation and cell cycle progression, suggesting a role of NtWLIM2 in the activation of basal histone gene expression. Interestingly, both live cell and in vitro data support NtWLIM2 di/oligomerization. We propose that NtWLIM2 functions as an actin-stabilizing protein, which, upon cytoskeleton remodeling, shuttles to the nucleus in order to modify gene expression.

关键词： actin BY-2 cytoskeleton DNA-binding histone genes LIM Nicotiana tabacum promoter regulation trans-acting factors.

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Three-dimensional automatic artificial intelligence driven augmented-reality selective biopsy during nerve-sparing robot-assisted radical prostatectomy:A feasibility and accuracy study

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Asian Journal of Urology 2023年第4期10卷 407-415页

作者： Enrico Checcucci Alberto Piana Gabriele Volpi Pietro Piazzolla Daniele Amparore sabrina De Cillis Federico Piramide Cecilia gatti Ilaria Stura Enrico Bollito Federica Massa Michele Di Dio Cristian Fiori Francesco Porpiglia Department of Surgery Candiolo Cancer InstituteFPO-IRCCSCandioloTurinItaly Department of Oncology Division of UrologyUniversity of TurinSan Luigi Gonzaga HospitalOrbassanoToItaly Department of Mechanical Engineering Politecnico di MilanoMilanItaly Department of Public Health and Pediatric Sciences School of MedicineUniversity of TurinItaly Department of Pathology San Luigi Gonzaga HospitalUniversity of TurinOrbassanoItaly SS Annunziata Hospital Department of SurgeryDivision of UrologyCosenzaItaly

Objective:To evaluate the accuracy of our new three-dimensional(3D)automatic augmented reality(AAR)system guided by artificial intelligence in the identification of tumour’s location at the level of the preserved neurovascular bundle(NVB)at the end of the extirpative phase of nerve-sparing robot-assisted radical ***:In this prospective study,we enrolled patients with prostate cancer(clinical stages cT1ce3,cN0,and cM0)with a positive index lesion at target biopsy,suspicious for capsular contact or extracapsular extension at preoperative multiparametric magnetic resonance *** underwent robot-assisted radical prostatectomy at San Luigi Gonzaga Hospital(Orbassano,Turin,Italy),from December 2020 to December *** the end of extirpative phase,thanks to our new AAR artificial intelligence driven system,the virtual prostate 3D model allowed to identify the tumour’s location at the level of the preserved NVB and to perform a selective excisional biopsy,sparing the remaining portion of the *** and postoperative data were evaluated,especially focusing on the positive surgical margin(PSM)rates,potency,continence recovery,and biochemical ***:Thirty-four patients were *** 15(44.1%)cases,the target lesion was in contact with the prostatic capsule at multiparametric magnetic resonance imaging(Wheeler grade L2)while in 19(55.9%)cases extracapsular extension was detected(Wheeler grade L3).3D AAR guided biopsies were negative in all pathological tumour stage 2(pT2)patients while they revealed the presence of cancer in 14 cases in the pT3 cohort(14/16;87.5%).PSM rates were 0%and 7.1%in the pathological stages pT2 and pT3(<3 mm,Gleason score 3),***:With the proposed 3D AAR system,it is possible to correctly identify the lesion’s location on the NVB in 87.5%of pT3 patients and perform a 3D-guided tailored nerve-sparing even in locally advanced diseases,without compromising the oncological safety

关键词： Prostate cancer Augmented reality Artificial intelligence Robotics Radical prostatectomy

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