Publications by authors named "Marco Nardini"

131 Publications

Truncated (2/2) hemoglobin: Unconventional structures and functional roles in vivo and in human pathogenesis.

Mol Aspects Med 2021 Nov 11:101049. Epub 2021 Nov 11.

Department of Biosciences, University of Milano, Milano, Italy. Electronic address:

Truncated hemoglobins (trHbs) build a sub-class of the globin family, found in eubacteria, cyanobacteria, unicellular eukaryotes, and in higher plants; among these, selected human pathogens are found. The trHb fold is based on a 2/2 α-helical sandwich, consisting of a simplified and reduced-size version of the classical 3/3 α-helical sandwich of vertebrate and invertebrate globins. Phylogenetic analysis indicates that trHbs further branch into three groups: group I (or trHbN), group II (or trHbO), and group III (or trHbP), each group being characterized by specific structural features. Among these, a protein matrix tunnel, or a cavity system implicated in diatomic ligand diffusion through the protein matrix, is typical of group I and group II, respectively. In general, a highly intertwined network of hydrogen bonds stabilizes the heme bound ligand, despite variability of the heme distal residues in the different trHb groups. Notably, some organisms display genes from more than one trHb group, suggesting that trHbN, trHbO, and trHbP may support different functions in vivo, such as detoxification of reactive nitrogen and oxygen species, respiration, oxygen storage/sensoring, thus aiding survival of an invading microorganism. Here, structural features and proposed functions of trHbs from human pathogens are reviewed.
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http://dx.doi.org/10.1016/j.mam.2021.101049DOI Listing
November 2021

The USR domain of USF1 mediates NF-Y interactions and cooperative DNA binding.

Int J Biol Macromol 2021 Dec 18;193(Pt A):401-413. Epub 2021 Oct 18.

Dipartimento di Bioscienze, Università degli Studi di Milano, Milano 20133, Italy. Electronic address:

The trimeric CCAAT-binding NF-Y is a "pioneer" Transcription Factor -TF- known to cooperate with neighboring TFs to regulate gene expression. Genome-wide analyses detected a precise stereo-alignment -10/12 bp- of CCAAT with E-box elements and corresponding colocalization of NF-Y with basic-Helix-Loop-Helix (bHLH) TFs. We dissected here NF-Y interactions with USF1 and MAX. USF1, but not MAX, cooperates in DNA binding with NF-Y. NF-Y and USF1 synergize to activate target promoters. Reconstruction of complexes by structural means shows independent DNA binding of MAX, whereas USF1 has extended contacts with NF-Y, involving the USR, a USF-specific amino acid sequence stretch required for trans-activation. The USR is an intrinsically disordered domain and adopts different conformations based on E-box-CCAAT distances. Deletion of the USR abolishes cooperative DNA binding with NF-Y. Our data indicate that the functionality of certain unstructured domains involves adapting to small variation in stereo-alignments of the multimeric TFs sites.
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http://dx.doi.org/10.1016/j.ijbiomac.2021.10.056DOI Listing
December 2021

High Conformational Flexibility of the E2F1/DP1/DNA Complex.

J Mol Biol 2021 09 25;433(18):167119. Epub 2021 Jun 25.

Dipartimento di Bioscienze, Università degli Studi di Milano, Via Celoria 26, 20133 Milano, Italy. Electronic address:

The E2F1 transcription factor is a master regulator of cell-cycle progression whose uncontrolled activation contributes to tumor cells growth. E2F1 binds DNA as a heterodimer with DP partners, resulting in a multi-domain quaternary-structure complex composed of DNA binding domains, a coiled coil domain and a marked box domain separated by short linkers. Building on the 3D knowledge of the single domains of E2F and DPs, we characterized the structure and dynamics of the complete E2F1/DP1/DNA complex by a combination of small-angle X-ray scattering and molecular dynamics simulations. It shows an asymmetric contribution of the dynamics of the two proteins. Namely, the coiled-coil domain leans toward the DP1 side of the complex; the DP1 loop between α2 and α3 of the DBD partially populates a helical structure leaning far from the DNA and in the same direction of the coiled-coil domain; and the N-terminal disordered region of DP1, rich in basic residues, contributes to DNA binding stabilization. Intriguingly, tumor mutations in the flexible regions of the complex suggest that perturbation of protein dynamics could affect protein function in a context-dependent way. Our data suggest fundamental contributions of DP proteins in distinct aspects of E2F biology.
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http://dx.doi.org/10.1016/j.jmb.2021.167119DOI Listing
September 2021

Minimally Invasive Plication of the Diaphragm: A Single-Center Prospective Study.

Innovations (Phila) 2021 Jul-Aug;16(4):343-349. Epub 2021 Jun 15.

156705 Department of Thoracic Surgery, James Cook University Hospital, Middlesbrough, UK.

Objective: Plication of the diaphragm is a life-changing procedure for patients affected by diaphragm paralysis. Traditionally, this procedure is performed through a thoracotomy. Access to the diaphragm via this incision is poor and the indications for surgery are limited to patients who can actually sustain such an invasive approach and associated morbidities. A minimally invasive approach was developed to improve the surgical management of diaphragm paralysis.

Methods: Patients underwent minimally invasive diaphragm plication either by video-assisted or robotic surgery through a 3-port technique with CO insufflation. Patients were followed at the routine 6-week clinic and also by telephone consultation 6 to 12 months postoperatively. Data were collected on postoperative complications, postoperative pain or numbness, symptomatic improvement, and change to quality of life following surgery.

Results: Forty-eight patients underwent 49 minimally invasive diaphragm plication. Median postoperative length of hospital stay was 4 days (range: 2 to 34 days) and there were no cases of mortality. Mean reduction in Medical Research Council dyspnea score per patient was 2.2 points (mode: 3 points). Twenty-eight patients (77.8%) reported a significant symptomatic improvement enabling improvements in quality of life, and 97.2% ( = 35) were satisfied with the surgical outcome.

Conclusions: Minimally invasive diaphragm plication is a safe procedure associated with prompt postoperative recovery. It is effective at reducing debilitating dyspnea and improving quality of life.
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http://dx.doi.org/10.1177/15569845211011583DOI Listing
November 2021

The activity and stability of a cold-active acylaminoacyl peptidase rely on its dimerization by domain swapping.

Int J Biol Macromol 2021 Jun 26;181:263-274. Epub 2021 Mar 26.

Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, 20126 Milan, Italy. Electronic address:

The study of enzymes from extremophiles arouses interest in Protein Science because of the amazing solutions these proteins adopt to cope with extreme conditions. Recently solved, the structure of the psychrophilic acyl aminoacyl peptidase from Sporosarcina psychrophila (SpAAP) pinpoints a mechanism of dimerization unusual for this class of enzymes. The quaternary structure of SpAAP relies on a domain-swapping mechanism involving the N-terminal A1 helix. The A1 helix is conserved among homologous mesophilic and psychrophilic proteins and its deletion causes the formation of a monomeric enzyme, which is inactive and prone to aggregate. Here, we investigate the dimerization mechanism of SpAAP through the analysis of chimeric heterodimers where a protomer lacking the A1 helix combines with a protomer carrying the inactivated catalytic site. Our results indicate that the two active sites are independent, and that a single A1 helix is sufficient to partially recover the quaternary structure and the activity of chimeric heterodimers. Since catalytically competent protomers are unstable and inactive unless they dimerize, SpAAP reveals as an "obligomer" for both structural and functional reasons.
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http://dx.doi.org/10.1016/j.ijbiomac.2021.03.150DOI Listing
June 2021

Producing natural vanilla extract from green vanilla beans using a β-glucosidase from Alicyclobacillus acidiphilus.

J Biotechnol 2021 Mar 27;329:21-28. Epub 2021 Jan 27.

University of Nottingham, School of Chemistry, University Park, Nottingham, NG7 2RD, United Kingdom; University of Bern, Department of Chemistry, Biochemistry, Pharmaceutical Sciences, Freiestrasse 3, Bern, CH-3012, Switzerland. Electronic address:

Current methods for the production of natural vanilla extract are long and tedious, and the efficiency of the vanillin extraction is usually conditioned by different factors during the traditional curing process (temperatures and weather conditions). As an important fraction of vanillin is present in the form of glucovanillin in green beans, endogenous β-glucosidases contribute to its hydrolysis; however, these enzymes lose efficiency during the curing process. The use of extremophilic organisms as a source of an appropriate exogenous enzyme can offer a valid alternative when producing natural vanillin. Here, a β-glucosidase from the thermo-acidophilic organism Alicyclobacillus acidiphilus (AacGH1) was cloned, expressed in E. coli BL21, and fully characterized in respect to both function and crystal structure. Notably, AacGH1 was stable at a temperature up to 50 °C and exhibited good tolerance to glucose, fructose and organic solvents, in particular it maintained full activity in the presence of up to 20 % (v/v) ethanol. The enzyme was then successfully applied to an ethanol-water (20 % (v/v)) extract of green vanilla beans and the complete hydrolysis of glucovanillin (1.7 mM) to vanillin, and other flavour compounds commonly found in vanilla, was achieved using 0.5 mg/mL of enzyme in just 15 min at 30 °C.
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http://dx.doi.org/10.1016/j.jbiotec.2021.01.017DOI Listing
March 2021

Structural Basis of Inhibition of the Pioneer Transcription Factor NF-Y by Suramin.

Cells 2020 10 29;9(11). Epub 2020 Oct 29.

Department of Biosciences, University of Milano, Via Celoria 26, 20133 Milano, Italy.

NF-Y is a transcription factor (TF) comprising three subunits (NF-YA, NF-YB, NF-YC) that binds with high specificity to the CCAAT sequence, a widespread regulatory element in gene promoters of prosurvival, cell-cycle-promoting, and metabolic genes. Tumor cells undergo "metabolic rewiring" through overexpression of genes involved in such pathways, many of which are under NF-Y control. In addition, NF-YA appears to be overexpressed in many tumor types. Thus, limiting NF-Y activity may represent a desirable anti-cancer strategy, which is an ongoing field of research. With virtual-screening docking simulations on a library of pharmacologically active compounds, we identified suramin as a potential NF-Y inhibitor. We focused on suramin given its high water-solubility that is an important factor for in vitro testing, since NF-Y is sensitive to DMSO. By electrophoretic mobility shift assays (EMSA), isothermal titration calorimetry (ITC), STD NMR, X-ray crystallography, and molecular dynamics (MD) simulations, we showed that suramin binds to the histone fold domains (HFDs) of NF-Y, preventing DNA-binding. Our analyses, provide atomic-level detail on the interaction between suramin and NF-Y and reveal a region of the protein, nearby the suramin-binding site and poorly conserved in other HFD-containing TFs, that may represent a promising starting point for rational design of more specific and potent inhibitors with potential therapeutic applications.
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http://dx.doi.org/10.3390/cells9112370DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7692634PMC
October 2020

Structural determinants for NF-Y subunit organization and NF-Y/DNA association in plants.

Plant J 2021 01 27;105(1):49-61. Epub 2020 Nov 27.

Dipartimento di Bioscienze, Università degli Studi di Milano, Via Celoria 26, Milano, 20133, Italy.

NF-Y transcription factor comprises three subunits: NF-YA, NF-YB and NF-YC. NF-YB and NF-YC dimerize through their histone fold domain (HFD), which can bind DNA in a non-sequence-specific fashion while serving as a scaffold for NF-YA trimerization. Upon trimerization, NF-YA specifically recognizes the CCAAT box sequence on promoters and enhancers. In plants, each NF-Y subunit is encoded by several genes giving rise to hundreds of potential heterotrimeric combinations. In addition, plant NF-YBs and NF-YCs interact with other protein partners to recognize a plethora of genomic motifs, as the CCT protein family that binds CORE sites. The NF-Y subunit organization and its DNA-binding properties, together with the NF-Y HFD capacity to adapt different protein modules, represent plant-specific features that play a key role in development, growth and reproduction. Despite their relevance, these features are still poorly understood at the molecular level. Here, we present the structures of Arabidopsis and rice NF-YB/NF-YC dimers, and of an Arabidopsis NF-Y trimer in complex with the FT CCAAT box, together with biochemical data on NF-Y mutants. The dimeric structures identify the key residues for NF-Y HFD stabilization. The NF-Y/DNA structure and the mutation experiments shed light on HFD trimerization interface properties and the NF-YA sequence appetite for the bases flanking the CCAAT motif. These data explain the logic of plant NF-Y gene expansion: the trimerization adaptability and the flexible DNA-binding rules serve the scopes of accommodating the large number of NF-YAs, CCTs and possibly other NF-Y HFD binding partners and a diverse audience of genomic motifs.
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http://dx.doi.org/10.1111/tpj.15038DOI Listing
January 2021

Structural and functional properties of Antarctic fish cytoglobins-1: Cold-reactivity in multi-ligand reactions.

Comput Struct Biotechnol J 2020 12;18:2132-2144. Epub 2020 Aug 12.

Institute of Biosciences and BioResources (IBBR), CNR, Via Pietro Castellino 111 80131 Napoli, Italy.

While the functions of the recently discovered cytoglobin, ubiquitously expressed in vertebrate tissues, remain uncertain, Antarctic fish provide unparalleled models to study novel protein traits that may arise from cold adaptation. We report here the spectral, ligand-binding and enzymatic properties (peroxynitrite isomerization, nitrite-reductase activity) of cytoglobin-1 from two Antarctic fish, and and present the crystal structure of cytoglobin-1. The Antarctic cytoglobins-1 display high O affinity, scarcely compatible with an O-supply role, a slow rate constant for nitrite-reductase activity, and do not catalyze peroxynitrite isomerization. Compared with mesophilic orthologues, the cold-adapted cytoglobins favor binding of exogenous ligands to the -coordinated bis-histidyl species, a trait related to their higher rate constant for distal-His/heme-Fe dissociation relative to human cytoglobin. At the light of a remarkable 3D-structure conservation, the observed differences in ligand-binding kinetics may reflect Antarctic fish cytoglobin-1 specific features in the dynamics of the heme distal region and of protein matrix cavities, suggesting adaptation to functional requirements posed by the cold environment. Taken together, the biochemical and biophysical data presented suggest that in Antarctic fish, as in humans, cytoglobin-1 unlikely plays a role in O transport, rather it may be involved in processes such as NO detoxification.
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http://dx.doi.org/10.1016/j.csbj.2020.08.007DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7451756PMC
August 2020

Imaging patterns of early stage lung cancer for the thoracic surgeon.

J Thorac Dis 2020 Jun;12(6):3349-3356

1Section of Thoracic Surgery, Department of General Surgery and Medical Specialities, 2Section of Radiology, Department of Medical Surgical Sciences and Advanced Technologies "GF Ingrassia", Policlinico University Hospital, Catania, Italy.

In the modern era, thoracic surgeons are experiencing an increase interest in imaging patterns of early stage lung cancer due to the introduction of the ground glass opacity in clinical practice, and for the necessity to an accurate cancer localization to perform the appropriate type of resection. In this brief review we analyze the latest news regarding imaging patterns of early pulmonary nodules with special emphasis to ground glass opacity.
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http://dx.doi.org/10.21037/jtd.2020.02.61DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7330749PMC
June 2020

Pulmonary nodules precision localization techniques.

Future Oncol 2020 Jun 21;16(16s):15-19. Epub 2020 May 21.

Department of Cardiothoracic Surgery, James Cook University Hospital, Middlesbrough, United Kingdom.

The advent of helical high-resolution CT scanners, the application of screening programs and the follow-up of patient with oncological history, led to an increasing number of diagnosis of small pulmonary nodule (less than 10 mm in maximum diameter), partially solid nodule or completely ground glass ones. Their management is controversial. Excisional biopsy by mean of video-assisted thoracic surgery is often a viable choice but to locate these lesions intraoperatively can be impossible without the aid of preoperative or intraoperative localization techniques. In this brief review we will analyze the benefit of adopting localization techniques prior to pulmonary resection for small pulmonary lesions and face the advantages and problems with the main techniques described in the literatures.
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http://dx.doi.org/10.2217/fon-2019-0069DOI Listing
June 2020

The co-existence of cold activity and thermal stability in an Antarctic GH42 β-galactosidase relies on its hexameric quaternary arrangement.

FEBS J 2021 01 4;288(2):546-565. Epub 2020 Jun 4.

Department of Biosciences, University of Milano, Italy.

To survive in cold environments, psychrophilic organisms produce enzymes endowed with high specific activity at low temperature. The structure of these enzymes is usually flexible and mostly thermolabile. In this work, we investigate the structural basis of cold adaptation of a GH42 β-galactosidase from the psychrophilic Marinomonas ef1. This enzyme couples cold activity with astonishing robustness for a psychrophilic protein, for it retains 23% of its highest activity at 5 °C and it is stable for several days at 37 °C and even 50 °C. Phylogenetic analyses indicate a close relationship with thermophilic β-galactosidases, suggesting that the present-day enzyme evolved from a thermostable scaffold modeled by environmental selective pressure. The crystallographic structure reveals the overall similarity with GH42 enzymes, along with a hexameric arrangement (dimer of trimers) not found in psychrophilic, mesophilic, and thermophilic homologues. In the quaternary structure, protomers form a large central cavity, whose accessibility to the substrate is promoted by the dynamic behavior of surface loops, even at low temperature. A peculiar cooperative behavior of the enzyme is likely related to the increase of the internal cavity permeability triggered by heating. Overall, our results highlight a novel strategy of enzyme cold adaptation, based on the oligomerization state of the enzyme, which effectively challenges the paradigm of cold activity coupled with intrinsic thermolability. DATABASE: Structural data are available in the Protein Data Bank database under the accession number 6Y2K.
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http://dx.doi.org/10.1111/febs.15354DOI Listing
January 2021

Mycobacterial and Human Nitrobindins: Structure and Function.

Antioxid Redox Signal 2020 08;33(4):229-246

Dipartimento di Scienze, Università Roma Tre, Roma, Italy.

Nitrobindins (Nbs) are evolutionary conserved all-β-barrel heme-proteins displaying a highly solvent-exposed heme-Fe(III) atom. The physiological role(s) of Nbs is almost unknown. Here, the structural and functional properties of ferric Nb (-Nb(III)) and ferric Nb (-Nb(III)) have been investigated and compared with those of ferric Nb (-Nb(III), nitrophorins (-NP(III)s), and mammalian myoglobins. Data here reported demonstrate that -Nb(III), -Nb(III), and -Nb(III) share with -NP(III)s the capability to bind selectively nitric oxide, but display a very low reactivity, if any, toward histamine. Data obtained overexpressing -Nb in human embryonic kidney 293 cells indicate that -Nb localizes mainly in the cytoplasm and partially in the nucleus, thanks to a nuclear localization sequence encompassing residues Glu124-Leu154. Human -Nb corresponds to the -terminal domain of the human nuclear protein THAP4 suggesting that Nb may act as a sensor possibly modulating the THAP4 transcriptional activity residing in the -terminal region. Finally, we provide strong evidence that both -Nb(III) and -Nb(III) are able to scavenge peroxynitrite and to protect free l-tyrosine against peroxynitrite-mediated nitration. Data here reported suggest an evolutionarily conserved function of Nbs related to their role as nitric oxide sensors and components of antioxidant systems. Human THAP4 may act as a sensing protein that couples the heme-based Nb(III) reactivity with gene transcription. -Nb(III) seems to be part of the pool of proteins required to scavenge reactive nitrogen and oxygen species produced by the host during the immunity response.
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http://dx.doi.org/10.1089/ars.2019.7874DOI Listing
August 2020

Structure and kinetic properties of human d-aspartate oxidase, the enzyme-controlling d-aspartate levels in brain.

FASEB J 2020 01 29;34(1):1182-1197. Epub 2019 Nov 29.

Dipartimento di Biotecnologie e Scienze della Vita, Università degli studi dell'Insubria, Varese, Italy.

d-Amino acids are the "wrong" enantiomers of amino acids as they are not used in proteins synthesis but evolved in selected functions. On this side, d-aspartate (d-Asp) plays several significant roles in mammals, especially as an agonist of N-methyl-d-aspartate receptors (NMDAR), and is involved in relevant diseases, such as schizophrenia and Alzheimer's disease. In vivo modulation of d-Asp levels represents an intriguing task to cope with such pathological states. As little is known about d-Asp synthesis, the only option for modulating the levels is via degradation, which is due to the flavoenzyme d-aspartate oxidase (DASPO). Here we present the first three-dimensional structure of a DASPO enzyme (from human) which belongs to the d-amino acid oxidase family. Notably, human DASPO differs from human d-amino acid oxidase (attributed to d-serine degradation, the main coagonist of NMDAR) showing peculiar structural features (a specific active site charge distribution), oligomeric state and kinetic mechanism, and a higher FAD affinity and activity. These results provide useful insights into the structure-function relationships of human DASPO: modulating its activity represents now a feasible novel therapeutic target.
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http://dx.doi.org/10.1096/fj.201901703RDOI Listing
January 2020

The structural bases for agonist diversity in an glutamate receptor-like channel.

Proc Natl Acad Sci U S A 2020 01 23;117(1):752-760. Epub 2019 Dec 23.

Department of Biosciences, University of Milan, 20133 Milan, Italy;

glutamate receptor-like (GLR) channels are amino acid-gated ion channels involved in physiological processes including wound signaling, stomatal regulation, and pollen tube growth. Here, fluorescence microscopy and genetics were used to confirm the central role of GLR3.3 in the amino acid-elicited cytosolic Ca increase in seedling roots. To elucidate the binding properties of the receptor, we biochemically reconstituted the GLR3.3 ligand-binding domain (LBD) and analyzed its selectivity profile; our binding experiments revealed the LBD preference for l-Glu but also for sulfur-containing amino acids. Furthermore, we solved the crystal structures of the GLR3.3 LBD in complex with 4 different amino acid ligands, providing a rationale for how the LBD binding site evolved to accommodate diverse amino acids, thus laying the grounds for rational mutagenesis. Last, we inspected the structures of LBDs from nonplant species and generated homology models for other GLR isoforms. Our results establish that GLR3.3 is a receptor endowed with a unique amino acid ligand profile and provide a structural framework for engineering this and other GLR isoforms to investigate their physiology.
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http://dx.doi.org/10.1073/pnas.1905142117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6955363PMC
January 2020

Precision surgery in lung metastasectomy.

Future Oncol 2020 Jun 20;16(16s):7-13. Epub 2019 Dec 20.

Service of Thoracic Surgery, University Hospital of Catania, Italy.

The value of pulmonary metastasis (PM) resection in the context of controlled primary tumor sites was shown to improve survival of patients if complete resection could be achieved. The surgeon's approach can be modulated by various parameters pertaining to safety margins including local growth properties, size, spread and location of PMs. Lymph node dissection and assessment is recommended although its impact on survival remains unclear. Thoracoscopic surgery combined to thin slice chest CT scans has shown results comparable to thoracotomy in patients with few PMs. The management of PMs should therefore be discussed on an individual, interdisciplinary basis to offer the best possible oncological and surgical results as well as to maximize long term patient survival rates.
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http://dx.doi.org/10.2217/fon-2018-0713DOI Listing
June 2020

Structural and Functional Characterization of the Globin-Coupled Sensors of and .

Antioxid Redox Signal 2020 02 3;32(6):378-395. Epub 2019 Dec 3.

Department of Biomedical Sciences, University of Antwerp, Wilrijk, Belgium.

Structural and functional characterization of the globin-coupled sensors (GCSs) from (GReg) and (GReg). Ultraviolet/visible and resonance Raman spectroscopies confirm the presence in GReg and GReg of a globin domain capable of reversible gaseous ligand binding. In GReg, an influence of the transmitter domain on the heme proximal region of the globin domain can be seen, and ' is higher than for other GCSs. The O binding kinetics suggests the presence of an open and a closed conformation. As for GReg, the fully oxygenated GReg show a very high diguanylate cyclase activity. The carbon monoxide rebinding to GReg indicates that intra- and intermolecular interactions influence the ligand binding. The globin domains of both proteins (GReg globin domain and GRegGb with cysteines (Cys16, 45, 114, 154) mutated to serines [GReg-Gb*]) share the same GCS fold, a similar proximal but a different distal side structure. They homodimerize through a G-H helical bundle as in other GCSs. However, GReg-Gb* shows also a second dimerization mode. This article extends our knowledge on the GCS proteins and contributes to a better understanding of the GCSs role in the formation of bacterial biofilms. GReg and GReg conform to the GCS family, share a similar overall structure, but they have different properties in terms of the ligand binding. In particular, GReg shows an open and a closed conformation that in the latter form will very tightly bind oxygen. GReg has only one closed conformation. In both proteins, it is the fully oxygenated GCS form that catalyzes the production of the second messenger.
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http://dx.doi.org/10.1089/ars.2018.7690DOI Listing
February 2020

Does cytoreduction surgery and hyperthermic intrathoracic chemotherapy prolong survival in patients with N0-N1 nonsmall cell lung cancer and malignant pleural effusion?

Eur Respir Rev 2019 Sep 31;28(153). Epub 2019 Jul 31.

Dipartimento di Specialita Medico-Chirurgiche, Universita degli Studi di Catania, Catania, Italy.

Introduction: Debulking surgery and hyperthermic intrathoracic chemotherapy (HITHOC) has been successfully used in the treatment of thoracic tumours. Few authors report on the feasibility of its use in patients with lung cancer and malignant pleural effusion. The aim of this study was to evaluate the efficacy and results of debulking surgery and HITHOC in the treatment of selected patients with nonsmall cell lung cancer (NSCLC) and malignant pleural effusion.

Methods: A systematic review was conducted in MEDLINE in accordance with PRISMA guidelines. The word search included: "hyperthermic intrathoracic chemotherapy and/or HITHOC or hyperthermic intrapleural". Inclusion criteria were only those studies reporting a sufficient amount of data on HITHOC and surgery for lung cancer. Single case reports and review articles were excluded.

Results: 20 articles were selected as they related to the topic of HITHOC and lung cancer. Most were from China (n=8) and Japan (n=6). Only four out of the 20 articles had sufficient data for this review. In total, data for 21 patients were collected. Debulking surgery ranged from wedge resection to pneumonectomy and pleurectomy. Mean survival was 27 months and median survival was 18 months (range 1-74 months). 13 patients out of 21 (62%) were alive at 1 year and six (28.5%) were alive at 2 years. 10 patients were still alive at the time of the respective publication in the 21 patients included. Systemic toxicity and treatment-related mortality were nil. There were insufficient data to perform a meta-analysis.

Conclusion: Although reported survival in this systematic review is encouraging, available evidence concerning debulking surgery and HITHOC in N0-N1 NSCLC with malignant pleural effusion is weak. Better evidence in the form of a randomised controlled trial is mandatory.
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http://dx.doi.org/10.1183/16000617.0018-2019DOI Listing
September 2019

Subxiphoid robotic-assisted right pneumonectomy.

J Thorac Dis 2019 Apr;11(4):1629-1631

Department of Cardiothoracic, James Cook University Hospital, Middlesbrough, UK.

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http://dx.doi.org/10.21037/jtd.2019.03.63DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6531753PMC
April 2019

Higher Number of Night Shifts Associates with Good Perception of Work Capacity and Optimal Lung Function but Correlates with Increased Oxidative Damage and Telomere Attrition.

Biomed Res Int 2019 11;2019:8327629. Epub 2019 Apr 11.

Department of Medical Sciences, University of Ferrara, Ferrara, Italy.

Sleep deprivation and the consequent circadian clock disruption has become an emergent health question being associated with premature aging and earlier chronic diseases onset. Night-shift work leads to circadian clock misalignment, which is linked to several age-related diseases. However, mechanisms of this association are not well understood. Aim of this study is to explore in night-shift workers early indicators of oxidative stress response and biological aging [oxidized/methylated DNA bases and leukocytes telomere length (LTL)] and late indicators of functional aging [lung function measurements (FEV1 and FVC)] in relation to personal evaluation of work capacity, measured by work ability index (WAI). One hundred fifty-five hospital workers were studied within the framework of a cross-sectional study. We collected physiological, pathological, and occupational history including pack-years, alcohol consumption, physical activity, and night shifts, together with blood and urine samples. Relationships were appraised by univariate and multivariate ordered-logistic regression models. We found that workers with good and excellent WAI present higher FEV1 (p< 0.01) and number of night-work shifts (p<0.05), but they reveal higher urinary levels of 8-oxoGua (p<0.01) and shorter LTL (p<0.05). We confirmed that higher work ability was prevalent among chronological younger workers (p<0.05), who have also a significant reduced number of diseases, particularly chronic (p<0.01) and musculoskeletal diseases (p<0.01). The new findings which stem from our work are that subjects with the highest work ability perception may have more demanding and burdensome tasks; they in fact present the highest number of night-shift work and produce unbalanced oxidative stress response that might induce premature aging.
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http://dx.doi.org/10.1155/2019/8327629DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6487156PMC
November 2019

Medical student exposure to cardiothoracic surgery in the United Kingdom.

Interact Cardiovasc Thorac Surg 2019 Mar 15. Epub 2019 Mar 15.

Department of Cardiothoracic Surgery, James Cook University Hospital, Middlesbrough, UK.

Objectives: There has been declining interest in cardiothoracic surgery amongst medical graduates. This survey examines the exposure of British medical students to cardiothoracic surgery in various settings and its relationship with students' interest in the speciality.

Methods: A questionnaire composed of 14 quantitative and qualitative items was distributed amongst 162 medical students. The survey included questions on demographics, interest in cardiothoracic surgery, mechanisms of exposure to the speciality and desire to pursue a career in cardiothoracic surgery before and after exposure.

Results: Amongst the surveyed students, 71.0% reported exposure to cardiothoracic surgery as part of their medical school curricula and 24.7% reported extracurricular exposure. Of the students, 46.7% reported clinical exposure. Overall, 27.1% of students reported interest in a career in cardiothoracic surgery, which was higher amongst students who had curricular (29.6%), clinical (35.5%) or extracurricular exposure (50.0%). Amongst interested students, 43.2% engaged in extracurricular cardiothoracic activities compared with 16.1% of students not interested in pursuing the speciality. Confidence in career choice after exposure increased more in interested students (20.4%) than not interested students (1.6%). Students rated exposure and mentorship as the most important factor in promoting a career in cardiothoracic surgery.

Conclusions: Medical students with an interest in cardiothoracic surgery are more likely to organize independent attachments in the speciality and attend extracurricular events; however, many students might fail to identify cardiothoracic surgery as an area of interest because of the lack of exposure at medical school.
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http://dx.doi.org/10.1093/icvts/ivz038DOI Listing
March 2019

The Reamer-Irrigator-Aspirator technique for manubriosternal non-union repair†.

Interact Cardiovasc Thorac Surg 2019 Mar 8. Epub 2019 Mar 8.

Department of Cardiothoracic Surgery, James Cook University Hospital, Middlesbrough, UK.

Autologous bone graft is used in the treatment of fracture non-unions. A novel approach to treat painful manubriosternal non-unions is described with bone graft harvested from the femur and plating.
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http://dx.doi.org/10.1093/icvts/ivz001DOI Listing
March 2019

Abiraterone acetate exerts a cytotoxic effect in human prostate cancer cell lines.

Naunyn Schmiedebergs Arch Pharmacol 2019 06 15;392(6):729-742. Epub 2019 Feb 15.

Section of Pharmacology, Department of Molecular and Translational Medicine, University of Brescia, V.le Europa 11, 25123, Brescia, Italy.

To study the capability of the CYP17A1 inhibitor abiraterone acetate (AER) to antagonize the androgen receptor (AR) activation in human prostate cancer (PCa) cell lines. T877A-AR-LNCaP, WT-AR-VCaP, AR-negative DU145, and PC3 PCa cell lines were used by MTT and cell count to study the ability of AER and enzalutamide (ENZ) to modify cell viability. The role of ARs in LNCaP was demonstrated through a gene-silencing experiment. The mechanism of AER cytotoxicity in LNCaP cells was studied, as well as the ability of AER to modulate AR gene expression. The in silico docking approach was applied to study the interaction of AER and ENZ with T877A-AR. Through high-performance liquid chromatography, the production of the AER main metabolite Δ4A was studied. AER bound AR in an almost identical manner to that of dihydrotestosterone (DHT). The higher binding energy for AER in T877A-AR could explain the major cytotoxic effect observed in LNCaP cells. The capability of LNCaP cells to synthesize Δ4A could mediate, at least in part, this effect. AER cytotoxicity in LNCaP cells was mainly due to the activation of apoptosis. Further, AER induced modification of AR target gene expression, suggesting a direct effect on AR activity. AER-induced cytotoxicity on PCa cell lines seemed to be mediated by binding with AR. The higher affinity of AER for T877A-AR may suggest a potential role of AER in the management of CRPC carrying this mutation; however, T877A-AR expressing CRPC patients developed AER resistance, probably due to the increase of progesterone.
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http://dx.doi.org/10.1007/s00210-019-01622-5DOI Listing
June 2019

The phosphorylatable Ser320 of NF-YA is involved in DNA binding of the NF-Y trimer.

FASEB J 2019 04 27;33(4):4790-4801. Epub 2018 Dec 27.

Dipartimento di Bioscienze, Università degli Studi di Milano, Milan, Italy.

Nuclear factor Y (NF-Y) is a transcription factor trimer binding to the functionally important CCAAT box, present in promoters of growth-promoting and cell cycle-regulated genes. The regulatory nuclear factor YA (NF-YA) subunit confers sequence-specificity to the histone-like nuclear factor YB/YC dimer. NF-YA harbors 2 serines-Ser320 and Ser326-shown to be phosphorylated by cyclin-dependent kinase 2. High-throughput proteomics data indicate that they are phosphorylated in vivo. Specifically, Ser320 makes structural contacts with the DNA phosphate backbone; Ser320-P is the major NF-YA phosphorylation isoform following overexpression in HeLa cells, increasing upon mitotic arrest. EMSA with recombinant Ala and Glu mutants confirm a role of Ser320, but not Ser326, in stabilization of DNA binding. Transactivation assays of the CCAAT-dependent MDR1 and RHOB promoters show loss in transcription function for Ser320Glu and Ser320Ala NF-YA mutants. Phylogenetic analysis of NF-YA proteins indicates that Ser320 is indeed evolutionarily conserved. We conclude that phosphorylation of this residue belongs to the core mechanisms of DNA-binding control, possibly driven by the necessity to unfasten binding of or to evict NF-Y from CCAAT sites under specific conditions of growth regulation.-Bernardini, A., Lorenzo, M., Nardini, M., Mantovani, R., Gnesutta, N. The phosphorylatable Ser320 of NF-YA is involved in DNA binding of the NF-Y trimer.
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http://dx.doi.org/10.1096/fj.201801989RDOI Listing
April 2019

Predicting return to work after acute myocardial infarction: Socio-occupational factors overcome clinical conditions.

PLoS One 2018 13;13(12):e0208842. Epub 2018 Dec 13.

Department of Medical Sciences, University of Ferrara, Ferrara, Italy.

Objectives: Return to work after acute myocardial infarction (AMI), a leading cause of death globally, is a multidimensional process influenced by clinical, psychological, social and occupational factors, the single impact of which, however, is still not well defined. The objective of this study was to investigate these 4 factors on return to work (RTW) within 365 days after AMI in a homogeneous cohort of patients who had undergone an urgent coronary angioplasty.

Participants: We studied 102 patients, in employment at the time of AMI (88.24% of men), admitted to the Department of Cardiology of the University-Hospital of Ferrara between March 2015 to December 2016. Demographical and clinical characteristics were obtained from the cardiological records. After completing an interview on social and occupational variables and the Hospital Anxiety and Depression (HADS) questionnaire, patients underwent exercise capacity measurement and spirometry.

Results: Of the 102 patients, only 12 (12.76%) held a university degree, 68.63% were employees and 31.37% self-employed. The median number of sick-leave days was 44 (IQR 33-88). At day 30, 78.5% of all subjects had not returned to work, at day 60, 40.8% and at day 365 only 7.3% had not resumed working. At univariate analyses, educational degree (p = 0.026), self-employment status (p = 0.0005), white collar professional category (p = 0.020) and HADS depression score were significant for earlier return to work. The multivariate analysis confirms that having a university degree, being self-employed and presenting a lower value of HADS depression score increase the probability of a quicker return to work.

Conclusions: These findings suggest that the strongest predictors of returning to work within 1 year after discharge for an acute myocardial infarction are related more to socio-occupational than to clinical parameters.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0208842PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6292571PMC
May 2019

Extended pleurectomy decortication: the current role.

Transl Lung Cancer Res 2018 Oct;7(5):556-561

Department of Thoracic Surgery, St Bartholomew's Hospital, London, UK.

Extended pleurectomy/decortication (EPD) has been formally defined but there remain technical areas of debate between practitioners. This is partly attributable to the relative rarity of this operation which is largely confined to a small number of specialist centres. Nevertheless, there is a widespread acceptance that extended pleurectomy/decortication (P/D) is a realistic and favourable alternative to extrapleural pneumonectomy. There may, however, remain a small number of clinical cases where this more extensive operation may be indicated. Preservation of the lung has widened the selection criteria for this form of radical mesothelioma surgery but there remain important factors to consider when offering extended P/D. In many patients with poorer prognostic factors the less radical operation of video assisted partial pleurectomy may be preferable. However, a randomized trial showed no survival benefit for this operation over simple talc pleurodesis. The future for P/D may also lie in the outcome of the MARS2 randomized controlled trial which will report in the next few years. Meanwhile there is a clinical and ethical dilemma when asked to perform this operation outside of the context of a clinical trial in the face of the lack of high grade evidence. The role of P/D is in one respect expanding but this may be short lived pending the findings of its assessment against non-surgical treatment.
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http://dx.doi.org/10.21037/tlcr.2018.06.07DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6204414PMC
October 2018

Identification of Plasmodium berghei Oocyst Rupture Protein 2 (ORP2) domains involved in sporozoite egress from the oocyst.

Int J Parasitol 2018 12 2;48(14):1127-1136. Epub 2018 Nov 2.

Foundation for Research and Technology-Hellas, Institute of Molecular Biology and Biotechnology, N. Plastira 100, GR 700 13 Heraklion, Greece. Electronic address:

Sporozoites are the infective form of malaria parasites which are transmitted from the mosquito salivary glands to a new host in a mosquito blood meal. The sporozoites develop inside the sporogonic oocyst and it is crucial for the continuation of the life cycle that the oocyst ruptures to release sporozoites. We recently described two Plasmodium Oocyst Rupture Proteins (ORP1 and ORP2), localized at the oocyst capsule, that are each essential for rupture of the oocysts. Both ORPs contain a histone fold domain implicated in the mechanism of oocyst rupture, possibly through the formation of a heterodimer between the two histone fold domains. To gain an understanding of the function of the different regions of the ORP2 protein, we generated deletion mutants. We monitored oocyst formation and rupture as well as sporozoites in the salivary gland. Our results show that different regions of ORP2 play independent roles in sporozoite egress. Deleting the N-terminal histone fold domain of ORP2 blocked sporozoite egress from the oocyst. Progressive deletions from the C-terminal resulted in no or significantly impaired sporozoite egress.
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http://dx.doi.org/10.1016/j.ijpara.2018.09.004DOI Listing
December 2018

Saturn-Shaped Ice Burst Pattern and Fast Basal Binding of an Ice-Binding Protein from an Antarctic Bacterial Consortium.

Langmuir 2019 06 26;35(23):7337-7346. Epub 2018 Sep 26.

Institute of Biochemistry, Food Science and Nutrition, The Robert H. Smith Faculty of Agriculture, Food and Environment , The Hebrew University of Jerusalem , Rehovot 7610001 , Israel.

Ice-binding proteins (IBPs) bind to ice crystals and control their growth, enabling host organisms to adapt to subzero temperatures. By binding to ice, IBPs can affect the shape and recrystallization of ice crystals. The shapes of ice crystals produced by IBPs vary and are partially due to which ice planes the IBPs are bound to. Previously, we have described a bacterial IBP found in the metagenome of the symbionts of Euplotes focardii ( EfcIBP). EfcIBP shows remarkable ice recrystallization inhibition activity. As recrystallization inhibition of IBPs and other materials are important to the cryopreservation of cells and tissues, we speculate that the EfcIBP can play a future role as an ice recrystallization inhibitor in cryopreservation applications. Here we show that EfcIBP results in a Saturn-shaped ice burst pattern, which may be due to the unique ice-plane affinity of the protein that we elucidated using the fluorescent-based ice-plane affinity analysis. EfcIBP binds to ice at a speed similar to that of other moderate IBPs (5 ± 2 mM s); however, it is unique in that it binds to the basal and previously unobserved pyramidal near-basal planes, while other moderate IBPs typically bind to the prism and pyramidal planes and not basal or near-basal planes. These insights into EfcIBP allow a better understanding of the recrystallization inhibition for this unique protein.
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http://dx.doi.org/10.1021/acs.langmuir.8b01914DOI Listing
June 2019

Forced expiratory volume in one second: A novel predictor of work disability in subjects with suspected obstructive sleep apnea.

PLoS One 2018 19;13(7):e0201045. Epub 2018 Jul 19.

Department of Medical Sciences, University of Ferrara, Ferrara, Italy.

Whether the association of work disability with obstructive sleep apnea (OSA) is mainly due to the disease, i.e. the number and frequency of apneas-hypoapneas, or to coexisting factors independent from the disease, is not well-established. In this study, we aim to evaluate work ability in a group of subjects undergoing OSA workup and to identify the major contributors of impaired work ability. In a cross-sectional study, we enrolled 146 consecutive subjects who have been working for the last five years and referred to the sleep disorders outpatients' clinic of the University-Hospital of Ferrara, Italy, with suspected OSA. After completing an interview in which the Work Ability Index (WAI) and the Epworth Sleepiness Scale (ESS) questionnaires were administered to assess work ability and excessive daytime sleepiness, respectively, subjects underwent overnight polysomnography for OSA diagnosing and spirometry. Of the 146 subjects, 140 (96%) completed the tests and questionnaires and, of these, 66 exhibited work disability (WAI < 37). OSA was diagnosed (apnea-hypopnea index ≥ 5) in 45 (68%) of the 66 subjects. After controlling for confounders, a lower level of forced expiratory volume at 1 second (FEV1), [odds ratio 0.97 (95% CI 0.95-1.00)], older age [1.09 (95% CI 1.03-1.15)], excessive daytime sleepiness [3.16 (95% CI 1.20-8.34)] and a worse quality of life [0.96 (95% CI 0.94-1.00)], but not OSA [1.04 (95% CI 0.41-2.62)], were associated with work disability. Patients with a higher number of diseases, in which OSA was not included, and a lower quality of life had an increased probability of absenteeism in the previous 12 months. In subjects with suspected OSA, FEV1 can be an important predictor of work disability.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0201045PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6053206PMC
January 2019

Robotic resection of a middle mediastinal mass.

J Vis Surg 2018 25;4:113. Epub 2018 May 25.

Department of Cardiothoracic Surgery, New York University Langone Health, New York, USA.

Aorto-pulmonary paraganglioma is an exceptionally rare condition, and its diagnosis and treatment are a challenge for the general thoracic surgeon. We describe the case of a 35 years old man who was incidentally diagnosed with a visceral mediastinal mass, deeply encased in the aorto-pulmonary window. To our knowledge this is the first case of its kind to be successfully treated with the adoption of a minimally invasive technique. We conclude that the dissection was made easier by the robotic instrumentation and by the camera system, and a minimally invasive approach would have been more difficult by traditional thoracoscopy.
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http://dx.doi.org/10.21037/jovs.2018.05.13DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5994472PMC
May 2018
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