The Protein Disulfide Isomerase gene family in bread wheat (T. aestivum L.)

<p>Abstract</p> <p>Background</p> <p>The Protein Disulfide Isomerase (PDI) gene family encodes several PDI and PDI-like proteins containing thioredoxin domains and controlling diversified metabolic functions, including disulfide bond formation and isomerisation during protein folding. Genomic, cDNA and promoter sequences of the three homoeologous wheat genes encoding the "typical" PDI had been cloned and characterized in a previous work. The purpose of present research was the cloning and characterization of the complete set of genes encoding PDI and PDI like proteins in bread wheat (<it>Triticum aestivum </it>cv Chinese Spring) and the comparison of their sequence, structure and expression with homologous genes from other plant species.</p> <p>Results</p> <p>Eight new non-homoeologous wheat genes were cloned and characterized. The nine PDI and PDI-like sequences of wheat were located in chromosome regions syntenic to those in rice and assigned to eight plant phylogenetic groups. The nine wheat genes differed in their sequences, genomic organization as well as in the domain composition and architecture of their deduced proteins; conversely each of them showed high structural conservation with genes from other plant species in the same phylogenetic group. The extensive quantitative RT-PCR analysis of the nine genes in a set of 23 wheat samples, including tissues and developmental stages, showed their constitutive, even though highly variable expression.</p> <p>Conclusions</p> <p>The nine wheat genes showed high diversity, while the members of each phylogenetic group were highly conserved even between taxonomically distant plant species like the moss <it>Physcomitrella patens</it>. Although constitutively expressed the nine wheat genes were characterized by different expression profiles reflecting their different genomic organization, protein domain architecture and probably promoter sequences; the high conservation among species indicated the ancient origin and diversification of the still evolving gene family. The comprehensive structural and expression characterization of the complete set of <it>PDI </it>and <it>PDI</it>-like wheat genes represents a basis for the functional characterization of this gene family in the hexaploid context of bread wheat.</p

Stratification of the risk of developing severe or lethal Covid-19 using a new score from a large Italian population: A population-based cohort study

Objectives To develop a population-based risk stratification model (COVID-19 Vulnerability Score) for predicting severe/fatal clinical manifestations of SARS-CoV-2 infection, using the multiple source information provided by the healthcare utilisation databases of the Italian National Health Service. Design Retrospective observational cohort study. Setting Population-based study using the healthcare utilisation database from five Italian regions. Participants Beneficiaries of the National Health Service, aged 18-79 years, who had the residentship in the five participating regions. Residents in a nursing home were not included. The model was built from the 7 655 502 residents of Lombardy region. Main outcome measure The score included gender, age and 29 conditions/diseases selected from a list of 61 conditions which independently predicted the primary outcome, that is, severe (intensive care unit admission) or fatal manifestation of COVID-19 experienced during the first epidemic wave (until June 2020). The score performance was validated by applying the model to several validation sets, that is, Lombardy population (second epidemic wave), and the other four Italian regions (entire 2020) for a total of about 15.4 million individuals and 7031 outcomes. Predictive performance was assessed by discrimination (areas under the receiver operating characteristic curve) and calibration (plot of observed vs predicted outcomes). Results We observed a clear positive trend towards increasing outcome incidence as the score increased. The areas under the receiver operating characteristic curve of the COVID-19 Vulnerability Score ranged from 0.85 to 0.88, which compared favourably with the areas of generic scores such as the Charlson Comorbidity Score (0.60). A remarkable performance of the score on the calibration of observed and predicted outcome probability was also observed. Conclusions A score based on data used for public health management accurately predicted the occurrence of severe/fatal manifestations of COVID-19. Use of this score may help health decision-makers to more accurately identify high-risk citizens who need early preventive or treatment interventions

Effect of resveratrol on mitochondrial function: Implications in parkin-associated familiar Parkinson's disease

Mitochondrial dysfunction and oxidative stress occur in Parkinson's disease (PD), but the molecular mechanisms controlling these events are not completely understood. Peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) is a transcriptional coactivator known as master regulator of mitochondrial functions and oxidative metabolism. Recent studies, including one from our group, have highlighted altered PGC-1α activity and transcriptional deregulation of its target genes in PD pathogenesis suggesting it as a new potential therapeutic target. Resveratrol, a natural polyphenolic compound proved to improve mitochondrial activity through the activation of several metabolic sensors resulting in PGC-1α activation. Here we have tested in vitro the effect of resveratrol treatment on primary fibroblast cultures from two patients with early-onset PD linked to different Park2 mutations. We show that resveratrol regulates energy homeostasis through activation of AMP-activated protein kinase (AMPK) and sirtuin 1 (SIRT1) and raise of mRNA expression of a number of PGC-1α's target genes resulting in enhanced mitochondrial oxidative function, likely related to a decrease of oxidative stress and to an increase of mitochondrial biogenesis. The functional impact of resveratrol treatment encompassed an increase of complex I and citrate synthase activities, basal oxygen consumption, and mitochondrial ATP production and a decrease in lactate content, thus supporting a switch from glycolytic to oxidative metabolism. Moreover, resveratrol treatment caused an enhanced macro-autophagic flux through activation of an LC3-independent pathway. Our results, obtained in early-onset PD fibroblasts, suggest that resveratrol may have potential clinical application in selected cases of PD-affected patients

Generation of tumour-specific cytotoxic T-cell clones from histocompatibility leucocyte antigen-identical siblings of patients with melanoma

Lymphodepletion and infusion of autologous expanded tumour-infiltrating lymphocytes is effective therapy for patients with malignant melanoma. Antitumour responses are likely to be mediated by HLA class I- and II-restricted immune responses directed at tumour antigens. We assessed whether the peripheral blood of normal HLA-matched siblings of patients with melanoma could be used to generate lymphocytes with antimelanoma activity for adoptive immunotherapy after allogeneic blood or marrow transplantation. Melanoma cell lines were derived from two donors and were used to stimulate the mononuclear cells of three HLA-identical siblings. CD4+ clones dominated cultures. Of these, approximately half were directly cytotoxic towards recipient melanoma cells and secreted interferon-γ in response to tumour stimulation. More than half of the noncytotoxic clones also secreted interferon-γ after melanoma stimulation. No CD4+ clones responded to stimulation with recipient haemopoietic cells. The majority of CD8+ clones directly lysed recipient melanoma, but did not persist in long-term culture in vitro. No crossreactivity with recipient haemopoietic cells was observed. The antigenic target of one CD4+ clone was determined to be an HLA-DR11-restricted MAGE-3 epitope. Antigenic targets of the remaining clones were not elucidated, but appeared to be restricted through a non-HLA-DR class II molecule. We conclude that the blood of allogeneic HLA-matched sibling donors contains melanoma-reactive lymphocyte precursors directed at tumour-associated antigens. Adoptive immunotherapy with unselected or ex vivo-stimulated donor lymphocytes after allogeneic stem cell transplantation has a rational basis for the treatment of malignant melanoma

Long-term outcome of COVID-19 patients treated with helmet noninvasive ventilation vs. high-flow nasal oxygen: a randomized trial

Background: Long-term outcomes of patients treated with helmet noninvasive ventilation (NIV) are unknown: safety concerns regarding the risk of patient self-inflicted lung injury and delayed intubation exist when NIV is applied in hypoxemic patients. We assessed the 6-month outcome of patients who received helmet NIV or high-flow nasal oxygen for COVID-19 hypoxemic respiratory failure. Methods: In this prespecified analysis of a randomized trial of helmet NIV versus high-flow nasal oxygen (HENIVOT), clinical status, physical performance (6-min-walking-test and 30-s chair stand test), respiratory function and quality of life (EuroQoL five dimensions five levels questionnaire, EuroQoL VAS, SF36 and Post-Traumatic Stress Disorder Checklist for the DSM) were evaluated 6&nbsp;months after the enrollment. Results: Among 80 patients who were alive, 71 (89%) completed the follow-up: 35 had received helmet NIV, 36 high-flow oxygen. There was no inter-group difference in any item concerning vital signs (N = 4), physical performance (N = 18), respiratory function (N = 27), quality of life (N = 21) and laboratory tests (N = 15). Arthralgia was significantly lower in the helmet group (16% vs. 55%, p = 0.002). Fifty-two percent of patients in helmet group vs. 63% of patients in high-flow group had diffusing capacity of the lungs for carbon monoxide &lt; 80% of predicted (p = 0.44); 13% vs. 22% had forced vital capacity &lt; 80% of predicted (p = 0.51). Both groups reported similar degree of pain (p = 0.81) and anxiety (p = 0.81) at the EQ-5D-5L test; the EQ-VAS score was similar in the two groups (p = 0.27). Compared to patients who successfully avoided invasive mechanical ventilation (54/71, 76%), intubated patients (17/71, 24%) had significantly worse pulmonary function (median diffusing capacity of the lungs for carbon monoxide 66% [Interquartile range: 47–77] of predicted vs. 80% [71–88], p = 0.005) and decreased quality of life (EQ-VAS: 70 [53–70] vs. 80 [70–83], p = 0.01). Conclusions: In patients with COVID-19 hypoxemic respiratory failure, treatment with helmet NIV or high-flow oxygen yielded similar quality of life and functional outcome at 6&nbsp;months. The need for invasive mechanical ventilation was associated with worse outcomes. These data indicate that helmet NIV, as applied in the HENIVOT trial, can be safely used in hypoxemic patients. Trial registration Registered on clinicaltrials.gov NCT04502576 on August 6, 202

Analisi di alcuni parametri nutrizionali in turioni di asparagi verdi (Asparagus officinalis)

Analysis of some nutritional parameters in green asparagus spears (asparagus officinalis) - lack of chemical and physical treatments post-harvest in green asparagus causes physiological and morphological changes after packaging and during storage. our aim was to monitor some nutritional parameters in the apical and basal portions of spear of the edible part (corresponding to 15 cm). after 9 days in the dark at 4 °c, there is an huge and slight decrease in ascorbate total content and soluble sugars, respectively. total chlorophyll and carotenoid levels decrease but the anthocyanins content not changes. Higher lignin level is mostly present in apical than basal portion

Argania spinosa (L.) Skeels: una risorsa per l’ambiente!

Argania spinosa (l.) Skeels, pianta spontanea del sudovest del marocco, è definita dai popoli berberi ‘albero della vita’ poiché ha un ruolo socio-economico ed ecologico importantissimo in questo ecosistema arido. il seme, da cui si estrae l’olio di argan, contenuto in una drupa dal colore giallastro a maturità, è ricco di saponine, polifenoli e acidi grassi insaturi (chaRRouf et al., 2007), sostanze dalle provate proprietà terapeutiche (dRiSSi et al., 2004; amzal et al., 2008). il legno della pianta è utilizzato come combustibile, i germogli dell’albero e i frutti rappresentano alimento per capre e cammelli. le piante di argania presentano, nel loro habitat naturale, una rilevante variabilità intraspecifica che si manifesta sia nella morfologia dei frutti che nella diversa produttività (bani-aameuR, 2002; alouani, bani-aameuR, 2004). Per questo motivo numerosi studi sono condotti con l’intento di selezionare genotipi altamente produttivi. nel campus dell’università di bari è presente un esemplare di argania che da diversi anni fiorisce e fruttifica abbondantemente e sui cui frutti sono state analizzate alcune caratteristiche della polpa. l’esame del colore, dimensioni, peso secco e fresco delle drupe ha permesso d’individuare, inizialmente, cinque stadi di maturazione su ognuno dei quali sono stati approfonditi alcuni aspetti fisiologici. il contenuto in zuccheri solubili quali glucosio, fruttosio e saccarosio, aumenta nel corso della maturazione; i contenuti di saccarosio e glucosio risultano più elevati in percentuale rispetto a quello in fruttosio. l’amido, dopo un picco nel secondo stadio di maturazione, diminuisce e il contenuto totale in carboidrati raggiunge il massimo nel terzo stadio di maturazione. il colore della parte carnosa delle drupe, è dovuto alla presenza dei pigmenti il cui rapporto quantitativo varia nel corso della maturazione. clorofilla a e clorofilla b diminuiscono con l’evolvere della maturazione mentre i carotenoidi aumentano nel terzo stadio di maturazione per poi declinare leggermente nel quarto e quinto stadio. con l’intento di individuare il momento della maturazione in cui si ha la massima espressione delle qualità nutrizionali della polpa di questi frutti, si è dosato il contenuto in acido ascorbico, un noto antiossidante largamente diffuso nelle piante. la forma ossidata dell’ acido ascorbico (acido deidroascorbico) non è stata mai riscontrata, mentre la forma ridotta è sempre presente, in maniera rilevante, pari a 0,2 mg g-1 di peso fresco, a partire dal terzo stadio di maturazione. dall’analisi globale dei dati, quello individuato come terzo stadio sembra quindi corrispondere alla fase di maturazione in cui vengono espresse al massimo alcune caratteristiche della parte carnosa del frutto quali il contenuto in zuccheri solubili, carotenoidi e acido ascorbico. gli ultimi due stadi, quarto e quinto, possono essere interpretati come un unico stadio in quanto privi di differenze significative morfo-fisiologiche. i dati ottenuti, anche se limitati ad un solo esemplare, possono rappresentare la base di partenza per individuare le peculiarità della pianta utili ad una migliore utilizzazione di questa risorsa ambientale. la polpa, sinora impiegata solo come materia residuale per alimentare capre e cammelli, nello stadio di maturazione più opportuno, quello definito come “terzo stadio”, potrebbe essere sfruttata come risorsa energetica (biofuel) e rappresentare così una ulteriore fattore di ricchezza della pianta

Le Corti regionali tra Stati e diritti. I sistemi di protezione dei diritti fondamentali europeo, americano e africano a confronto

I tre sistemi regionali di protezione dei diritti umani, europeo, americano ed africano, rappresentano, al contempo, una progressiva trascrizione della Dichiarazione Universale dei diritti dell\u2019uomo del 1948 ed una risposta al problema dell\u2019applicazione concreta dei diritti sulla base della prossimit\ue0 territoriale tra Stati che condividono una medesima area geopolitica. Tale processo ha raggiunto un alto grado di maturit\ue0 solamente nel sistema europeo; il sistema inter-americano risulta in fase di sviluppo; mentre il sistema africano, per la sua pi\uf9 recente affermazione, \ue8 chiaramente in una fase che si pu\uf2 definire embrionale. Cionondimeno il framework giuridico, le strutture istituzionali e il diritto sostanziale si presentano sotto molti profili omogenei. Al fine di mettere in evidenza differenze e similitudini tra i tre sistemi regionali, il volume propone una loro trattazione separata, muovendo dall\u2019analisi della declinazione del diritto alla vita. La vita, diritto a fondamento di tutti gli altri diritti, diventa un prisma attraverso cui analizzare l\u2019evoluzione di ciascuno dei tre sistemi, consentendo agli autori di interrogarsi ancora sul rapporto tra universalit\ue0 e tradizioni culturali. L\u2019idea di uomo che scaturisce dalla lettura delle norme convenzionali, e dall\u2019interpretazione fornitane dalle Corti regionali, mette alla prova sia l\u2019aspirazione ad un nucleo condiviso di tutela, sulla scia della Dichiarazione Universale, sia la ricerca delle specificit\ue0 regionali. Infine, Attraverso l\u2019analisi dei meccanismi di enformcement diretto ed indiretto presenti nei tre sistemi di tutela viene indagato il rapporto tra ordinamenti nazionali e Corti sovranazionali, per metterne in evidenza i successi e le tensioni interne ai tre modelli