765 Is acute response to calcium sensitizers drugs predictive of response to cardiac contractility modulation in NYHA IV patients?

Abstract A 53-years old man presented to our institution with a diagnosis of decompensated heart failure NYHA Class IV. He had a history of ischaemic heart disease with severe biventricular dysfunction, diabetes, hypertension, dyslipidaemia, advanced chronic kidney disease, previous explanation of dual-chamber implantable electronic device (ICD) due to endocarditis and subsequent implantation of subcutaneous ICD in primary prevention. Home therapy included uptitrated angiotensin-converting enzyme inhibitor, β-blocker, loop-diuretic, spironolactone, acetylsalicylic acid, and oral hypoglycemics. Clinical examination showed signs and symptoms of systemic and pulmonary congestion with pleural effusion and ascites. Echocardiography revealed diffuse left ventricular (LV) hypokinesis with an ejection fraction (EF) of 25%, severe right ventricular dysfunction and increased filling pressures. He was treated with high dose of i.v. diuretics with mild improvement of dyspnoea. However, haemodynamic stability was labile with worsening of symptoms as soon as mild down-titration of iv diuretics was attempted. Levosimendan, a calcium-sensitizer inodilator, indicated for short-term treatment of acutely decompensated severe chronic heart failure (HF), was administered with good clinical response. Thus, we thought that the patient could have benefited from contractility modulation therapy (CCM) which acts on intramyocardial calcium handling. CCM is a novel therapeutic option for patients with classes III–IV HF with EF ≥ 25% to ≤ 45% and narrow QRS complex that acts on intramyocardial calcium-handling. CCM proved effective in alleviating symptoms, improving exercise tolerance and quality of life, and reducing hospitalization rates in HF. It improves myocardial contractility, reverses the foetal myocyte gene program associated with HF and facilitates cardiac reverse remodelling. Therefore, an Optimizer Smart System (Impulse Dynamics) was implanted. Two pacing electrodes were placed on the interventricular septum in apical and mid-septal position, respectively. The leads were connected to a pulse-generator in a right pectoral pocket. In the following days, we observed a progressive improvement in clinical status, with gradual resolution of peripheral oedema, dyspnoea and fatigue and significant weight loss. Six-month echocardiography showed a stable value of EF and significant improvement in stroke volume (35.2 ml from 24.8 ml at baseline). The patient did not undergo further hospitalization for decompensated HF and was in stable ambulatory NYHA Class IV. We believe CCM is an option in patients with advanced HF in which avoiding recurrent hospitalizations, with their overt increase mortality, is often a challenging therapeutic goal. 765 Figur

Nextcast : A software suite to analyse and model toxicogenomics data

The recent advancements in toxicogenomics have led to the availability of large omics data sets, representing the starting point for studying the exposure mechanism of action and identifying candidate biomarkers for toxicity prediction. The current lack of standard methods in data generation and analysis hampers the full exploitation of toxicogenomics-based evidence in regulatory risk assessment. Moreover, the pipelines for the preprocessing and downstream analyses of toxicogenomic data sets can be quite challenging to implement. During the years, we have developed a number of software packages to address specific questions related to multiple steps of toxicogenomics data analysis and modelling. In this review we present the Nextcast software collection and discuss how its individual tools can be combined into efficient pipelines to answer specific biological questions. Nextcast components are of great support to the scientific community for analysing and interpreting large data sets for the toxicity evaluation of compounds in an unbiased, straightforward, and reliable manner. The Nextcast software suite is available at: ( https://github.com/fhaive/nextcast).(c) 2022 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).Peer reviewe

Effectiveness and safety of vedolizumab in a matched cohort of elderly and nonelderly patients with inflammatory bowel disease: the IG-IBD LIVE study

Background Vedolizumab registration trials were the first to include elderly patients with moderate-to-severe ulcerative colitis (UC) or Crohn's disease (CD), but few real-life data have been reported in this population. Aims We investigated the effectiveness and safety of vedolizumab in matched cohorts of elderly and nonelderly UC and CD patients. Methods The Long-term Italian Vedolizumab Effectiveness (LIVE) study is a retrospective-prospective study including UC and CD patients who started vedolizumab from April 2016 to June 2017. Elderly patients (>= 65 years) were matched clinically 1:2 to nonelderly patients (18-64 years); the 2 groups were followed until drug discontinuation or June 2019. Results The study included 198 elderly (108 UC, 90 CD) and 396 matched nonelderly patients (205 UC, 191 CD). Nonelderly UC patients had a significantly higher persistence on vedolizumab compared to elderly patients (67.6% vs. 51.4%, p = 0.02). No significant difference in effectiveness was observed between elderly and nonelderly CD patients (59.4% vs. 52.4%, p = 0.32). Age >= 65 years was associated with lower persistence in UC; for CD, previous exposure to anti-TNF-alpha agents, Charlson comorbidity index >2 and moderate-to-severe clinical activity at baseline were associated with lower persistence. There were recorded 130 adverse events, with comparable rates between the two groups. A Charlson comorbidity index >2 was associated with an increased risk of adverse events. Conclusion Vedolizumab can be considered a safe option in elderly IBD patients. Its effectiveness in elderly UC patients may be reduced, while no age-dependent effect on effectiveness was observed in CD

Molecular pathways shared between host-parasitoid interaction in insect and other animals: The case of teratocyte extracellular enolase

Molecular pathways shared between host-parasitoid interaction in insect and other animals: The case of teratocyte extracellular enolase Introduction: Enolase is an ubiquitous metalloenzyme involved in the glycolytic pathway. Interestingly it appears also as a multifunctional protein. Enolases anchored on the outer surface of the plasma membrane are involved in tissue invasion mechanisms by the activation of Plasminogen. Methods: To determine the Enolase localization and its capacity to bind and activate the plasminogen, experiments of immunogold labeling, indirect immunofluorescence staining, co-localization, binding and activation in vitro assay, Duolink in situ Proximity Ligation Assay (PLA) and dot blotting assay were carried out. Furthermore the Enolase ability to degrade extracellular matrix was in vitro evaluated by a Matrix digestion assay. Results/Conclusion: We have identified an extracellular Enolase (Ae-ENO) produced by teratocytes, embryonic cells of the insect parasitoid Aphidius ervi (Hymenoptera, Braconidae). For the first time we demonstrate that Ae-ENO, although lacking a signal peptide, accumulates in cytoplasmic vesicles oriented towards the cell membrane. Ae-ENO binds and activates a plasminogen-like molecule inducing digestion of the host tissue and thereby ensuring successful parasitism. The results support the hypothesis of the existence of plasminogen-like proteins in invertebrates and their activation through mechanisms involving the surface enolase/fibrinolytic system that until now seemed to be exclusively found in vertebrates, and that is conserved across species. These findings also suggest that Ae-ENO is a novel and unique example of gene sharing in insects

XXV CONGRESSO NAZIONALE ITALIANO DI ENTOMOLOGIA

L’enolasi extracellulare dei teratociti di Aphidius ervi (Ae-ENO) lega e attiva una proteina Plasminogen-like inducendo la degradazione della matrice extracellulare. Lo studio delle basi molecolari delle interazioni ospite-parassitoide nel sistema biologico Acyrthosiphon pisum / Aphidius ervi ha consentito di identificare i fattori parassitari di origine materna ed embrionale, che svolgono un ruolo importante nella regolazione dell’ospite. I teratociti, cellule derivanti dalla dissociazione della serosa embrionale del parassitoide, sono responsabili di una digestione extra-orale dei tessuti dell’ospite al fine di trasformarli in un substrato immediatamente disponibile per la progenie. Sono cellule che crescono rapidamente in dimensione, diventando altamente poliploidi, e il loro ruolo di supporto nutrizionale alla larva è sostenuto dalla capacità di sintetizzare e rilasciare nell’emolinfa in particolare due proteine: una Fatty Acid Binding Protein (Ae-FABP), che media il trasporto di acidi grassi nell’emolinfa dell’ospite per renderli disponibili alla larva, e una Enolasi (Ae-ENO). Quest’ultima, oltre alla normale attività di enzima glicolitico, è anche espressa sulla superficie esterna dei teratociti, sebbene la sua sequenza amminoacidica manchi del peptide segnale. Esperimenti di Immunogold labeling e Microscopia Elettronica a Trasmissione (TEM) hanno dimostrato che il trasporto dell’Ae-ENO verso la superficie dei teratociti potrebbe essere mediato da strutture esosoma-like, analogamente a quanto osservato in molti altri organismi eucarioti e procarioti. L’Ae-ENO localizzata sulla superficie cellulare dei teratociti funge da recettore di una proteina plasminogen-like presente nell’emolinfa dell’ospite A. pisum, trasformandola nella forma attiva, plasmina, responsabile della degradazione della matrice extracellulare (ECM) dell’ospite stesso. Esperimenti condotti in vitro hanno mostrato, infatti, che Ae-ENO presente sulla superficie dei teratociti interagisce con il plasminogeno umano attivandolo in plasmina in presenza del corrispondente attivatore uPA (Urokinase Plasminogen Activator). Inoltre, teratociti incubati con plasminogeno umano e uPA seminati su un supporto ECM-like, hanno dimostrato la loro capacità in vitro di degradazione della matrice extracellulare. Questi risultati supportano l'ipotesi dell’esistenza di proteine plasminogen-like negli invertebrati, la cui attivazione è mediata da un meccanismo che coinvolge una enolase extracellulare fino ad ora considerato esclusivo dei vertebrati, e che invece risulta essere conservato tra le specie. Si tratta quindi della prima dimostrazione di un processo di degradazione della ECM mediato dalla attivazione di una proteina Plasminogen-like presente nell’insetto ospite

Identification of major Toxoneuron nigriceps venom proteins using an integrated transcriptomic/proteomic approach

Endoparasitoids in the order Hymenoptera are natural enemies of several herbivorous insect pest species. During oviposition they inject a mixture of factors, which include venom, into the host, ensuring the successful parasitism and the development of their progeny. Although these parasitoid factors are known to be responsible for host manipulation, such as immune system suppression, little is known about both identity and function of the majority of their venom components. To identify the major proteins of Toxoneuron nigriceps (Hymenoptera: Braconidae) venom, we used an integrated transcriptomic and proteomic approach. The tandem-mass spectrometric (LC-MS/MS) data combined with T. nigriceps venom gland transcriptome used as a reference database resulted in the identification of a total of thirty one different proteins. While some of the identified proteins have been described in venom from several parasitoids, others were identified for the first time. Among the identified proteins, hydrolases constituted the most abundant family followed by transferases, oxidoreductases, ligases, lyases and isomerases. The hydrolases identified in the T. nigriceps venom glands included proteases, peptidases and glycosidases, reported as common components of venom from several parasitoid species. Taken together, the identified proteins included factors that could potentially inhibit the host immune system, manipulate host physiological processes and host development, as well as provide nutrients to the parasitoid progeny, degrading host tissues by specific hydrolytic enzymes. The venom decoding provides us with information about the identity of candidate venom factors which could contribute to the success of parasitism, together with other maternal and embryonic factors

Lentinus edodes Exposure before and after Fetus Implantation: Materno-Fetal Development in Rats with Gestational Diabetes Mellitus

Background: The presence of β-glucans and phenolic compounds in Lentinus edodes suggests this mushroom can be used as a nutritional supplement. Two gestational conditions (before and after fetus implantation) were evaluated, and Lentinus edodes exposure was performed in diabetes mellitus rat model induced by streptozotocin in pre-clinical tests. Methods: On the 20th day of pregnancy, cesarean sections were performed. Blood was collected for biochemical, hematologic parameters and oxidative stress biomarkers. Placenta and amniotic fluid were collected, and fetuses were analyzed through morphological evaluation. Results: The mushroom did not reduce the severe hyperglycemia of the mother-concept but promoted an increase in maternal insulin levels; reduced the levels of alanine aminotransferase, and aspartate aminotransferase, triglyceride and total cholesterol; protected the animals from post-implantation losses. Liver damage induced by streptozotocin was reversed in experimental groups. Conclusions: Lentinus edodes mushroom has antioxidant properties that can minimize the damage caused by gestational diabetes mellitus

XXV CONGRESSO NAZIONALE ITALIANO DI ENTOMOLOGIA

Identificazione delle principali componenti del veleno di Toxoneuron nigriceps integrando un approccio trascrittomico e proteomico. Toxoneuron nigriceps (V.) (Hymenoptera: Braconidae) è un endoparassitoide di stadi larvali di Heliothis virescens (F.) (Lepidoptera: Noctuidae). Al momento dell’ovideposizione, la femmina inietta nel corpo dell'ospite il veleno e il fluido del calice ovarico, un secreto denso contenente proteine ovariche e un polydnavirus del genere bracovirus (TnBV). Queste secrezioni materne, insieme a fattori embrionali, i teratociti, cellule derivanti dalla dissociazione della serosa, sono responsabili delle principali alterazioni fisiologiche osservate a carico dell’ospite, al fine di garantire un ambiente adeguato per lo sviluppo del parassitoide. Uno dei fattori chiave coinvolti nella regolazione dell’ospite, da parte del parassitoide, sembrerebbe essere il veleno. Ad oggi il ruolo del veleno di T. nigriceps risulta poco conosciuto, sebbene le informazioni preliminari indichino un coinvolgimento nella soppressione della risposta immunitaria. Le principali componenti del veleno di T. nigriceps sono state identificate mediante un approccio combinato di trascrittomica e proteomica. Il trascrittoma delle ghiandole del veleno è stato realizzato de novo mediante RNAseq e risulta costituito da 17742 contigs. Le sequenze sono state analizzate con il software Blast2GO fornendo un quadro completo delle putative proteine presenti nel veleno e le relative informazioni sulle funzioni molecolari, sui processi biologici e sui putativi compartimenti cellulari. L’analisi proteomica è stata condotta sulle componenti del veleno, mediante separazione per elettroforesi bidimensionale, escissione e digestione triptica degli spots e successiva analisi di spettrometria di massa tandem (HPLC-MS/MS). Il software MASCOT, ha consentito di incrociare le peak-list sperimentali con i dati di trascrittomica, permettendo di identificare numerose putative proteine presenti nel veleno di T. nigriceps. L’identificazione molecolare e la successiva caratterizzazione di queste molecole saranno essenziali per comprendere il ruolo svolto da ciascun componente nell’induzione e nella regolazione della sindrome patologica osservata negli ospiti parassitizzati, contribuendo a chiarire l'azione e l’interazione di tutti i fattori parassitari sia di origine materna che embrionale

The Toxoneuron nigriceps polydnavirus ANK1 protein induces apoptosis in insect S2 cells and Heliothis virescens hemocytes by interacting with Alix protein

The Toxoneuron nigriceps polydnavirus ANK1 protein induces apoptosis in insect S2 cells and Heliothis virescens hemocytes by interacting with Alix protein Intoduction: Toxoneuron nigriceps is an endophagous parasitoid of larval stages of tobacco budworm, Heliothis virescens. The bracovirus associated with this wasp (TnBV) is currently being studied, and several genes expressed in parasitized host larvae have been isolated and their possible role partly elucidated. In this work, we studied the role of the viral gene TnBVank1 in the apoptosis. TnBVank1 encodes an ankyrin motif protein similar to insect and mammalian IkB, inhibitor of the transcription nuclear factor kB. Methods: The role of TnBVank1 was investigated into H. virescens hemocytes by in vivo transient expression and, in vitro, in Drosophila Schneider's S2 cells which stably expressed TnBVank1. The success of in vivo and in vitro transfections was detected by RT-PCR and western blot. Apoptosis was detected by caspase-3 activity. ANK1 interactor proteins were reveled by coimmunoprecipitation experiments which indicated that ANK1 bound ALG-2- interacting protein X (Alix), an interactor of apoptosis-linked gene protein 2 (ALG-2). Alix was silenced by RNAi to study the function of ANK1-Alix interaction. Results/Conclusion: We show that when ANK1 was stably expressed in insect S2 cells, it stimulated caspase-3 activity. Furthermore, we get the same results into H. virescens hemocytes by in vivo transient expression of TnBVank1. Silencing Alix, ANK1 was no longer able to cause apoptosis in S2 cells and into H. virescens hemocytes. Collectively, these results indicate that ANK1 induces apoptosis by interacting with Alix, suggesting that this role could be relevant in the suppression of host immune response observed in tobacco budworm larvae parasitized by T. nigriceps

Development of new atherosclerotic plaque in hypertensive patients: an observational registry study from the Campania-Salute network

Carotid atherosclerotic plaques (CAPs) can develop despite appropriate antihypertensive therapy. In this observational study, we assessed characteristics associated with risk of incident CAP in a large hypertensive registry