Towards elucidating carnosic acid biosynthesis in Lamiaceae: Functional characterization of the three first steps of the pathway in Salvia fruticosa and Rosmarinus officinalis

Carnosic acid (CA) is a phenolic diterpene with anti-tumour, anti-diabetic, antibacterial and neuroprotective properties that is produced by a number of species from several genera of the Lamiaceae family, including Salvia fruticosa (Cretan sage) and Rosmarinus officinalis (Rosemary). To elucidate CA biosynthesis, glandular trichome transcriptome data of S. fruticosa were mined for terpene synthase genes. Two putative diterpene synthase genes, namely SfCPSand SfKSL, showing similarities to copalyl diphosphate synthase and kaurene synthase-like genes, respectively, were isolated and functionally characterized. Recombinant expression in Escherichia coli followed by in vitro enzyme activity assays confirmed that SfCPS is a copalyl diphosphate synthase. Coupling of SfCPS with SfKSL, both in vitro and in yeast, resulted in the synthesis miltiradiene, as confirmed by 1D and 2D NMR analyses (1H, 13C, DEPT, COSY H-H, HMQC and HMBC). Coupled transient in vivo assays of SfCPS and SfKSL in Nicotiana benthamiana further confirmed production of miltiradiene in planta. To elucidate the subsequent biosynthetic step, RNA-Seq data of S. fruticosa and R. officinalis were searched for cytochrome P450 (CYP) encoding genes potentially involved in the synthesis of the first phenolic compound in the CA pathway, ferruginol. Three candidate genes were selected, SfFS, RoFS1 and RoFS2. Using yeast and N. benthamiana expression systems, all three where confirmed to be coding for ferruginol synthases, thus revealing the enzymatic activities responsible for the first three steps leading to CA in two Lamiaceae genera

Genetic prediction of ICU hospitalization and mortality in COVID-19 patients using artificial neural networks

There is an unmet need of models for early prediction of morbidity and mortality of Coronavirus disease-19 (COVID-19). We aimed to a) identify complement-related genetic variants associated with the clinical outcomes of ICU hospitalization and death, b) develop an artificial neural network (ANN) predicting these outcomes and c) validate whether complement-related variants are associated with an impaired complement phenotype. We prospectively recruited consecutive adult patients of Caucasian origin, hospitalized due to COVID-19. Through targeted next-generation sequencing, we identified variants in complement factor H/CFH, CFB, CFH-related, CFD, CD55, C3, C5, CFI, CD46, thrombomodulin/THBD, and A Disintegrin and Metalloproteinase with Thrombospondin motifs (ADAMTS13). Among 381 variants in 133 patients, we identified 5 critical variants associated with severe COVID-19: rs2547438 (C3), rs2250656 (C3), rs1042580 (THBD), rs800292 (CFH) and rs414628 (CFHR1). Using age, gender and presence or absence of each variant, we developed an ANN predicting morbidity and mortality in 89.47% of the examined population. Furthermore, THBD and C3a levels were significantly increased in severe COVID-19 patients and those harbouring relevant variants. Thus, we reveal for the first time an ANN accurately predicting ICU hospitalization and death in COVID-19 patients, based on genetic variants in complement genes, age and gender. Importantly, we confirm that genetic dysregulation is associated with impaired complement phenotype.- Pfizer Pharmaceuticals(undefined

Detection of host genetic polymorphisms - biomarkers associated with viral respiratory infections severity

Influenza remains an important threat for human health, despite the extensive study of influenza viruses and the production of effective vaccines. In contrast to virus genetics determinants, host genetic factors with clinical impact remained unexplored until recently. The association between three single nucleotide polymorphisms (SNPs) and influenza outcome in a European population was investigated in the present study. All samples were collected during the influenza A(H1N1)pdm09 post-pandemic period 2010-11 and a sufficient number of severe and fatal cases was included. Host genomic DNA was isolated from pharyngeal samples of 110 patients from northern Greece with severe (n = 59) or mild (n = 51) influenza A(H1N1)pdm09 disease, at baseline, and the genotype of CD55 rs2564978, C1QBP rs3786054 and FCGR2A rs1801274 SNPs was investigated. Our findings suggest a relationship between the two complement-related SNPs, namely, the rare TT genotype of CD55 and the rare AA genotype of C1QBP with increased death risk. No significant differences were observed for FCGR2A genotypes neither with fatality nor disease severity. Additional large-scale genetic association studies are necessary for the identification of reliable host genetic risk factors associated with influenza A(H1N1)pdm09 outcome. Prophylactic intervention of additional high-risk populations, according to their genetic profile, will be a key achievement for the fight against influenza viruses.Οι ιογενείς αναπνευστικές λοιμώξεις, αποτέλεσαν και συνεχίζουν να αποτελούν σημαντικό παράγοντα νοσηρότητας και θνησιμότητας, παρά τα προγράμματα εντατικής επιτήρησης. Μετά και την πρόσφατη πανδημία γρίπης Α(Η1Ν1) 2009, έχει δοθεί ιδιαίτερη έμφαση στην κατανόηση των καθοριστικών παραγόντων και της παθογένεσης των σοβαρών αναπνευστικών λοιμώξεων. Το μεγαλύτερο μέρος της μέχρι τώρα βιβλιογραφίας έχει επικεντρωθεί στη μελέτη της γενετικής των ιών για την ανίχνευση των στοιχείων εκείνων που καθιστούν ορισμένα στελέχη ιδιαίτερα παθογονικά. Ωστόσο, απουσιάζουν επί του παρόντος βιβλιογραφικά δεδομένα που να περιγράφουν τους πιθανούς καθοριστικούς γενετικούς παράγοντες του ξενιστή που προκαλούν ευαισθησία σε λοίμωξη από αναπνευστικούς ιούς και μπορεί να οδηγήσουν σε σοβαρές αναπνευστικές παθολογικές καταστάσεις.Από τα 1847 δείγματα της μεταπανδημικής περιόδους 2010-11 που ελέγχθηκαν για τον ιό της γρίπης Α(Η1Ν1)pdm09 επιλέχθηκαν ύστερα από ορισμό κριτηρίων αποκλεισμού 110 δείγματα για γονοτυπικό έλεγχο και έλεγχο του λόγου των σχετικών πιθανοτήτων με την έκβαση της νόσου. Οι πολυμορφισμοί που μελετήθηκαν σχετίζονται με την ανοσιακή απάντηση και συγκεκριμένα είναι οι: CD55 rs2564978, C1QBP rs3786054 και FCGR2A rs1801274. Τα αποτελέσματά μας δείχνουν ότι οι πολυμορφισμοί CD55 rs2564978, C1QBP rs3786054 που σχετίζονται με το συμπλήρωμα, δηλαδή ο σπάνιος γονότυπος TT του CD55 καθώς και ο σπάνιος AA γονότυπος του C1QBP, συνδέονται με σημαντικά αυξημένο κίνδυνο θανάτου, αλλά όχι με τη σοβαρότητα της νόσου. Αντιθέτως δεν παρατηρήθηκαν σημαντικές διαφορές σε κανένα επίπεδο για τους γονοτύπους rs1801274 του FCGR2A γονιδίου.Με την αναγνώριση του ρόλου της γενετικής ποικιλότητας του ξενιστή, καθώς και με τον εντοπισμό των βασικών πολυμορφισμών που μειώνουν την ανοσιακή απάντηση ή σχετίζονται με την προστασία, θα είμαστε σε θέση να αναγνωρίζουμε υποπληθυσμούς/άτομα υψηλού κινδύνου οι οποίοι θα μπορούν να λάβουν προφυλακτικά μέτρα, αλλά θα οδηγήσουν και στη διασάφηση-αναγνώριση των κύριων πρωτεϊνών του ξενιστή που παίζουν σημαντικό ρόλο στην αλληλεπίδραση μεταξύ ιού και ξενιστή για τον καλύτερο σχεδιασμό φαρμακευτικών σκευασμάτων και εμβολίων

Retracted: Applications of Quantum Dots in Flow Cytometry

Non

CRISSPAC: A web-based platform for predicting the SYNTAX Score and severity of coronary artery disease

CRISSPAC is an open-source web-based platform for the prediction of the Syntax Score and the severity of coronary artery disease (CAD) providing a variety of data analytics and machine learning solutions presented via an-easy-to-use graphical interface environment. The aim of the software is two-fold: (1) to facilitate both retrospective and prospective objectives dedicated to the diagnosis of the complexity of CAD; (2) to support open science principles through the public availability of the source code. Our envision is to promote software extensibility and utilization towards accurate diagnosis, decision-making processes and personalized patient management and counseling activities