Morphological adaptation and variation of gene expression after pneumonectomy: observations on experimental animal model

Introduzione La pneumonectomia su modello animale potrebbe essere un’utile piattaforma di studio per approfondire i meccanismi della risposta compensatoria al danno polmonare. Scopo dello studio è determinare la presenza di variazioni morfologiche e di espressione del trascrittoma dopo pneumonectomia. Materiali e metodi Undici suini sono stati sottoposti a pneumonectomia sinistra. Sono stati eseguiti prelievi sito-specifici intraoperatori su polmone sinistro e successivamente confrontati con prelievi sito-specifici su polmone destro dopo eutanasia a 60 giorni. I prelievi degli animali con decorso regolare sono stati sottoposti a RNA-sequencing e successiva analisi computazionale per valutare il peso funzionale del singolo gene o di clusters di geni. Risultati Un animale è stato escluso per insorgenza di ernia diaframmatica. In 7/10 è stata riscontrata apertura della pleura mediastinica con parziale erniazione del polmone controlaterale e shift mediastinico. L’istologia ha mostrato dilatazione degli spazi aerei, rottura dei setti interalveolari, lieve infiammazione, assenza di fibrosi, stiramento radiale dei bronchi e riduzione del letto capillare. L’analisi di bulk RNA-sequencing ha identificato 553 geni espressi in modo differenziale (DEG)(P<0,001) tra pre e post-pneumonectomia. I primi 10 DEG up-regolati: Edn1, Areg, Havcr2, Gadd45g, Depp1, Cldn4, Atf3, Myc, Gadd45b, Socs3; i primi 10 geni down-regolati: Obscn, Cdkn2b, ENSSSCG00000015738, Prrt2, Amer1, Flrt3, Efnb2, Tox3, Znf793, Znf365. Tra i DEG è stata riscontrata una predominanza di geni specifici dei macrofagi. L’analisi di gene ontology basata su DAVID ha mostrato un significativo arricchimento della "via di segnalazione apoptotica estrinseca"(FDR q=7,60x10 -3), della via di “Risposta all’insulina”(FDR q=7,60x10 -3) ed un arricchimento di geni “Regolatori negativi del segnale DDX58/IFIH1”(FDR q=7.50x10 -4). Conclusioni Il presente studio conferma la presenza di variazioni macroscopiche e microscopiche fenotipiche dopo pneumonectomia. L’RNA sequencing e lo studio di genomica traslazionale hanno mostrato l’esistenza di geni singoli e di network di geni disregolati dopo pneumonectomia, prevalentemente in determinate popolazioni cellulari.Introduction Pneumonectomy on animal model could be useful to investigate mechanisms of the compensatory response to lung damage. Aim of the study is to determine the presence of morphological and transcriptome expression changes after pneumonectomy. Materials and methods Eleven pigs underwent left pneumonectomy. Intraoperative site-specific samples were performed on the left lung and subsequently compared with site-specific samples on the right lung after euthanasia at 60 days. The samples of the animals with a uneventful course were subjected to RNA-sequencing and subsequent computational analysis to evaluate the functional weight of the single gene or of clusters of genes. Results One animal was excluded for diaphragmatic hernia. In 7/10 mediastinal pleura was interrupted with partial herniation of the remaining lung and mediastinal shift. Histology showed dilation of the air spaces, rupture of the interalveolar septa, mild inflammation, absence of fibrosis, radial stretching of the bronchi and reduction of the capillary bed. Bulk RNA-sequencing analysis identified 553 differentially expressed genes (DEG) (P <0.001) between pre and post-pneumonectomy state. The top 10 up-regulated DEGs: Edn1, Areg, Havcr2, Gadd45g, Depp1, Cldn4, Atf3, Myc, Gadd45b, Socs3; the top 10 down-regulated genes: Obscn, Cdkn2b, ENSSSCG00000015738, Prrt2, Amer1, Flrt3, Efnb2, Tox3, Znf793, Znf365. A predominance of macrophage-specific genes was found among the DEGs. The DAVID-based gene ontology analysis showed a significant enrichment of the "extrinsic apoptotic signaling pathway" (FDR q = 7.60x10 -3), of the "Insulin response" pathway (FDR q = 7.60x10 - 3) and of “DDX58 / IFIH1 negative regulators” genes (FDR q = 7.50x10 -4). Conclusions The present study confirms the presence of macroscopic and microscopic phenotypic variations after pneumonectomy. This translational genomics study showed the existence of individual genes and networks of genes which resulted disregulated after pneumonectomy, mainly in certain cell populations

Dynamic titanium prosthesis based on 3D-printed replica for chest wall resection and reconstruction

3D-printing technologies can assist the surgical planning and prosthesis engineering for the management of extended chest wall resection. Different types of prosthesis have been utilized over time, but some concerns remain about their impact on the respiratory function. Here we present a new kind of 3D-printed titanium prosthesis designed to be either strong and flexible. The prosthesis was created on a 1:1 3D-printed anatomic replica of the chest, used to delineate surgical margins and to define the reconstructive requirements

Characterization of a new variant DNA (cytosine-5)-methyltransferase unable to methylate double stranded DNA isolated from the marine annelid worm Chaetopterus variopedatus

AbstractThe enzyme S-adenosylmethionine-DNA (cytosine-5)-methyltransferase has been identified, first time for invertebrates, in embryos of the marine polychaete annelid worm Chaetopterus variopedatus. The molecule has been isolated from embryos at 15 h of development. It is a single peptide of about 200 kDa molecular weight, cross-reacting with antibodies against sea urchin DNA methyltransferase. The enzymatic properties of the molecule are similar to those of Dnmt1 methyltransferases isolated from other organisms, but with the peculiarity to be unable to make ‘de novo’ methylation on double stranded DNA

Concomitant Intubation with Minimal Cuffed Tube and Rigid Bronchoscopy for Severe Tracheo-Carinal Obstruction

Background: Our aim was to report on the use of an innovative technique for airway management utilizing a small diameter, short-cuffed, long orotracheal tube for assisting operative rigid bronchoscopy in critical airway obstruction. Methods: We retrospectively reviewed the clinical data of 36 patients with life-threatening critical airway stenosis submitted for rigid bronchoscopy between January 2008 and July 2021. The supporting ventilatory tube, part of the Translaryngeal Tracheostomy KIT (Fantoni method), was utilized in tandem with the rigid bronchoscope during endoscopic airway reopening. Results: Indications for collateral intubation were either tumors of the trachea with near-total airway obstruction (13), or tumors of the main carina with total obstruction of one main bronchus and possible contralateral involvement (23). Preliminary dilation was necessary before tube placement in only 2/13 patients with tracheal-obstructing tumors (15.4%). No postoperative complications were reported. There was one case of an intraoperative cuff tear, with no further technical problems. Conclusions: In our experience, this innovative method proved to be safe, allowing for continuous airway control. It enabled anesthesia inhalation, use of neuromuscular blockage and reliable end-tidal CO2 monitoring, along with protection of the distal airway from blood flooding. The shorter time of the procedure was due to the lack of need for pauses to ventilate the patient

Release of DNA from Dermanyssus gallinae during the Biting Process

none10noDermanyssus gallinae is a hematophagous ectoparasitic mite that usually infests poultry, but is also known for occasionally attacking other animals and humans. It represents a major problem for poultry systems all over the world, with detrimental effects for both production and animal welfare. Despite the significance of D. gallinae, very little is known about the biting process to date. Therefore, this study has aimed to verify if mite DNA is injected into the host skin during the blood meal. Mite DNA has been detected by seminested PCR from infested chicken skin and quantified by real-time PCR. Furthermore, its localization within the host tissue has been checked by fluorescent in situ hybridization. Results showed that a very little amount of D. gallinae DNA can be released by mites, suggesting that the latter do not introduce whole or partially destroyed cells into the host, but rather it injects traces of nucleic acids, possibly together with merocrine secretions.Pugliese, Nicola; Raele, Donato Antonio; Schiavone, Antonella; Cafiero, Maria Assunta; Potenza, Lucia; Samarelli, Rossella; Circella, Elena; Vasco, Ilaria; Pennuzzi, Germana; Camarda, AntonioPugliese, Nicola; Raele, Donato Antonio; Schiavone, Antonella; Cafiero, Maria Assunta; Potenza, Lucia; Samarelli, Rossella; Circella, Elena; Vasco, Ilaria; Pennuzzi, Germana; Camarda, Antoni

Release of {DNA} from Dermanyssus gallinae during the Biting Process

: Dermanyssus gallinae is a hematophagous ectoparasitic mite that usually infests poultry, but is also known for occasionally attacking other animals and humans. It represents a major problem for poultry systems all over the world, with detrimental effects for both production and animal welfare. Despite the significance of D. gallinae, very little is known about the biting process to date. Therefore, this study has aimed to verify if mite DNA is injected into the host skin during the blood meal. Mite DNA has been detected by seminested PCR from infested chicken skin and quantified by real-time PCR. Furthermore, its localization within the host tissue has been checked by fluorescent in situ hybridization. Results showed that a very little amount of D. gallinae DNA can be released by mites, suggesting that the latter do not introduce whole or partially destroyed cells into the host, but rather it injects traces of nucleic acids, possibly together with merocrine secretions

Inhibition of ERK1/2 signaling prevents bone marrow fibrosis by reducing osteopontin plasma levels in a myelofibrosis mouse model

Clonal myeloproliferation and development of bone marrow (BM) fibrosis are the major pathogenetic events in myelofibrosis (MF). The identification of novel antifibrotic strategies is of utmost importance since the effectiveness of current therapies in reverting BM fibrosis is debated. We previously demonstrated that osteopontin (OPN) has a profibrotic role in MF by promoting mesenchymal stromal cells proliferation and collagen production. Moreover, increased plasma OPN correlated with higher BM fibrosis grade and inferior overall survival in MF patients. To understand whether OPN is a druggable target in MF, we assessed putative inhibitors of OPN expression in vitro and identified ERK1/2 as a major regulator of OPN production. Increased OPN plasma levels were associated with BM fibrosis development in the Romiplostim-induced MF mouse model. Moreover, ERK1/2 inhibition led to a remarkable reduction of OPN production and BM fibrosis in Romiplostim-treated mice. Strikingly, the antifibrotic effect of ERK1/2 inhibition can be mainly ascribed to the reduced OPN production since it could be recapitulated through the administration of anti-OPN neutralizing antibody. Our results demonstrate that OPN is a novel druggable target in MF and pave the way to antifibrotic therapies based on the inhibition of ERK1/2-driven OPN production or the neutralization of OPN activity

Prognostic Relevance of Multi-Antigenic Myeloma-Specific T-Cell Assay in Patients with Monoclonal Gammopathies

: Multiple Myeloma (MM) typically originates from underlying precursor conditions, known as Monoclonal Gammopathy of Undetermined Significance (MGUS) and Smoldering Multiple Myeloma (SMM). Validated risk factors, related to the main features of the clonal plasma cells, are employed in the current prognostic models to assess long-term probabilities of progression to MM. In addition, new prognostic immunologic parameters, measuring protective MM-specific T-cell responses, could help to identify patients with shorter time-to-progression. In this report, we described a novel Multi-antigenic Myeloma-specific (MaMs) T-cell assay, based on ELISpot technology, providing simultaneous evaluation of T-cell responses towards ten different MM-associated antigens. When performed during long-term follow-up (mean 28 months) of 33 patients with either MGUS or SMM, such deca-antigenic myeloma-specific immunoassay allowed to significantly distinguish between stable vs. progressive disease (p &lt; 0.001), independently from the Mayo Clinic risk category. Here, we report the first clinical experience showing that a wide (multi-antigen), standardized (irrespective to patients' HLA), MM-specific T-cell assay may routinely be applied, as a promising prognostic tool, during the follow-up of MGUS/SMM patients. Larger studies are needed to improve the antigenic panel and further explore the prognostic value of MaMs test in the risk assessment of patients with monoclonal gammopathies

Prognostic factors of lung cancer in lymphoma survivors (the LuCiLyS study)

Background Second cancer is the leading cause of death in lymphoma survivors, with lung cancer representing the most common solid tumor. Limited information exists about the treatment and prognosis of second lung cancer following lymphoma. Herein, we evaluated the outcome and prognostic factors of Lung Cancer in Lymphoma Survivors (the LuCiLyS study) to improve the patient selection for lung cancer treatment. Methods This is a retrospective multicentre study including consecutive patients treated for lymphoma disease that subsequently developed non-small cell lung cancer (NSCLC). Data regarding lymphoma including age, symptoms, histology, disease stage, treatment received and lymphoma status at the time of lung cancer diagnosis, and data on lung carcinoma as age, smoking history, latency from lymphoma, symptoms, histology, disease stage, treatment received, and survival were evaluated to identify the significant prognostic factors for overall survival. Results Our study population included 164 patients, 145 of which underwent lung cancer resection. The median overall survival was 63 (range, 58–85) months, and the 5-year survival rate 54%. At univariable analysis no-active lymphoma (HR: 2.19; P=0.0152); early lymphoma stage (HR: 1.95; P=0.01); adenocarcinoma histology (HR: 0.59; P=0.0421); early lung cancer stage (HR: 3.18; P&lt;0.0001); incidental diagnosis of lung cancer (HR: 1.71; P&lt;0.0001); and lung cancer resection (HR: 2.79; P&lt;0.0001) were favorable prognostic factors. At multivariable analysis, no-active lymphoma (HR: 2.68; P=0.004); early lung cancer stage (HR: 2.37; P&lt;0.0001); incidental diagnosis of lung cancer (HR: 2.00; P&lt;0.0001); and lung cancer resection (HR: 2.07; P&lt;0.0001) remained favorable prognostic factors. Patients with non-active lymphoma (n=146) versus those with active lymphoma (n=18) at lung cancer diagnosis presented better median survival (64 vs. 37 months; HR: 2.4; P=0.02), but median lung cancer specific survival showed no significant difference (27 vs. 19 months; HR: 0.3; P=0.17). Conclusions The presence and/or a history of lymphoma should not be a contraindication to resection of lung cancer. Inclusion of lymphoma survivors in a lung cancer-screening program may lead to early detection of lung cancer, and improve the survival

A Proposal for a Three Detector Short-Baseline Neutrino Oscillation Program in the Fermilab Booster Neutrino Beam

A Short-Baseline Neutrino (SBN) physics program of three LAr-TPC detectors located along the Booster Neutrino Beam (BNB) at Fermilab is presented. This new SBN Program will deliver a rich and compelling physics opportunity, including the ability to resolve a class of experimental anomalies in neutrino physics and to perform the most sensitive search to date for sterile neutrinos at the eV mass-scale through both appearance and disappearance oscillation channels. Using data sets of 6.6e20 protons on target (P.O.T.) in the LAr1-ND and ICARUS T600 detectors plus 13.2e20 P.O.T. in the MicroBooNE detector, we estimate that a search for muon neutrino to electron neutrino appearance can be performed with ~5 sigma sensitivity for the LSND allowed (99% C.L.) parameter region. In this proposal for the SBN Program, we describe the physics analysis, the conceptual design of the LAr1-ND detector, the design and refurbishment of the T600 detector, the necessary infrastructure required to execute the program, and a possible reconfiguration of the BNB target and horn system to improve its performance for oscillation searches.Comment: 209 pages, 129 figure