Thrombophilia and abdominal vessel thrombosis in a Greek University hospital: A five year experience

Thrombophilia, either congenital or acquired, has foremost consequences in the abdominal vessels. We review here the cases of patients admitted with abdominal vessel thrombosis over a five-year period. Our data focused on gender and age at diagnosis, site of thrombosis, previous thromboembolic events, underlying conditions, and family history. Investigations included measurement of protein C, protein S, activated PC resistance, and antithrombin, and screening for factor V Leiden, prothrombin G20210A, the C677T variant of the methylenetetrahydrofolate reductase gene, and V617F JAK2 mutation, r lupus anticoagulant, antiphospholipid antibodies and paroxysmal nocturnal hemospherinouria, and also serum folate, vitamin 12, and total homocysteine concentrations. Twenty-nine patients were admitted and 18 of their family members also underwent the same thrombophilia investigations. Eighteen patients (62.1%) presented with portal vein thrombosis (PVT), five patients (17.2%) with mesenteric vein thrombosis (MVT), four patients (13.8%) with splenic vein thrombosis (SVT), and two (6.9%) patients with hepatic vein thrombosis (HVT). There was a high incidence of congenital thrombophilia (37.9%), acquired thrombophilic conditions (27.6%), or both (20.7%). Sixteen of 18 patients with PVT, four of five patients with MVT, all four patients with SVT, and one of two patients with HVT had one or more thrombophilic risk factors. In 13.8% of the patients no underlying condition was identified. We concluded that thrombophilia may have major implications in the pathogenesis of abdominal vessel thrombosis in adult life, and a thorough thrombophilia investigation should be performed in all these patients

Molecular analysis of TP53 mutations and determination of p53 protein in patients with SCD receiving hydroxyurea over long time periods

The purpose of this study is the analysis of TP53 gene and the determination of p53 protein in patients with SCD, being under treatment with HU from 3-8 years. TP53 gene contributes to the reparation process of DNA as well as to apoptosis whereas its mutations consist one of the mayor and early events in the pathogenesis of malignant transformation. Its disorders are often accompanied by mutated protein production that is characterized by greater stability in relation to wild type protein. The identification of certain changes, occurring in the TP53 gene and the corresponding protein, could imply the possible mutagen or leukemogen action of HU. 51 patients with SCD are included in the study, who take HU since 1991 until today. Initially all 51 patients were studied following 0-62 months of treatment with median duration a 28,2-month period. 26 of them were re-examined during the term of 51-97 months of treatment with median duration an 68,1-month term. 10 of 51 patients were re-examined during the term of 31-43 months of treatment with median duration an 37,9-month term. The tracing of mutations in TP53 gene was based on the PCR- SSCP analysis. Following the extraction of RNA, and the production of cDNA, four DNA areas measuring a length of 257-302 Bp, in which exons 5-11 are included, were selectively enhanced with PCR. SSCP analysis followed, and the set off of single strand selected areas of DNA following the staining of gel with silver nitrate. The determination of p53 phosphoprotein in the serum of the patients was accomplished by means of the non-antagonistic Elisa, where the antigen was defined with the double antibody method. The SSCP analysis of TP53 in the 87 samples did not reveal disorders in the electrophoretic mobility, and consequently structural abnormalities of the selected single-strand regions. The quantitative determination of the p53 antigen in these patients with SCD and normal SSCP analysis of TP53, revealed antigen concentrations ranging within 1,5-20 pg/ml. On the contrary, the comparative quantitative determination of p53 patients with hematological malignancies of poor prognosis and pathological SSCP analysis of p53 gene, revealed p53 antigen concentrations ranging within 90-850 pg/ml. The non-identification of certain disorders of the TP53 gene and the p53 protein could characterize the term of 3-8 years of treatment with HU of the patients with SCD, as adequately safe as to its potentially mutagen or leukemogen action of HU.Το ΤΡ53 γονίδιο μετέχει στη διαδικασία επιδιόρθωσης του DNA και στην απόπτωση, ενώ οι μεταλλάξεις του αποτελούν ένα από τα κύρια και πρώιμα γεγονότα στη παθογένεση της κακοήθους εξαλλαγής. Οι διαταραχές του συνοδεύονται συχνά από την παραγωγή μεταλλαγμένης πρωτεΐνης που χαρακτηρίζεται από μεγαλύτερη σταθερότητα σε σχέση με τις φυσιολογικές. Η αναγνώριση συγκεκριμένων αλλαγών στο ΤΡ53 γονίδιο και την αντίστοιχη πρωτεΐνη θα μπορούσαν να υποδηλώνουν την πιθανή μεταλλαξιογόνο ή λευχαιμογόνο δράση της HU σε ασθενείς SCD. Στη μελέτη περιλαμβάνονται 51 ασθενείς με SCD που λαμβάνουν HU από το 1991 έως σήμερα. Αρχικά μελετήθηκαν και οι 51 ασθενείς μετά από 0-62 μήνες θεραπείας, και μέση διάρκεια τους 28,2 μήνες. Οι 26 από αυτούς επανελέγχθησαν κατά την περίοδο των 51-97 μηνών θεραπείας και μέση διάρκεια τους 68,1 μήνες. Οι 10 από τους 51 ασθενείς που είχαν ελεγχθεί πριν την έναρξη HU, επανελέγχθηκαν μετά από 31-43 μήνες αγωγής και μέση διάρκεια τους 37,9 μήνες. Η ανίχνευση των μεταλλάξεων στο ΤΡ53 γονίδιο βασίσθηκε στην SSCP ανάλυση. Μετά την απομόνωση του RNA και την παραγωγή cDNA, 4 περιοχές του cDNA μήκους 257-302 Bp στις οποίες περιέχονται τα εξόνια 5-11, εκλεκτικά ενισχύθηκαν με PCR. Ακολούθησε η SSCP ανάλυση και η ανάδειξη των μονόκλωνων επιλεγμένων περιοχών του DNA μετά από χρώση του πηκτώματος με νιτρικό άργυρο. Ο προσδιορισμός της p53 φωσφοπρωτεΐνης στον ορό των ασθενών έγινε με μη ανταγωνιστική ELISA όπου προσδιορίσθηκε το αντιγόνο με την μέθοδο του διπλού αντισώματος. Η SSCP ανάλυση του ΤΡ53 στα 77 δείγματα δεν ανέδειξε διαταραχές στην ηλεκτροφορητική κινητικότητα και συνεπώς στη στερεοδομή των επιλεγμένων μονόκλωνων περιοχών. Ο ποσοτικός προσδιορισμός του ΤΡ53 αντιγόνου σε 77 δείγματα από 51 ασθενείς με SCD και φυσιολογική SSCP ανάλυση του p53, ανέδειξε συγκεντρώσεις αντιγόνου από 1,5-20 pg/ml. Αντίθετα, ο συγκριτικός ποσοτικός προσδιορισμός του p53 αντιγόνου σε 8 ασθενείς με αιματολογικές κακοήθειες κακής πρόγνωσης και παθολογική SSCP ανάλυση του p53 γονιδίου, ανέδειξε συγκεντρώσεις αντιγόνου από 90-850 pg/ml. Η μη αναγνώριση συγκεκριμένων διαταραχών του ΤΡ53 γονιδίου και της p53 πρωτεΐνης θα μπορούσε να χαρακτηρίσει την περίοδο των 3-8 ετών αγωγής με HU των ασθενών με SCD, ως επαρκώς ασφαλή ως προς τη δυνητικά μεταλλαξιογόνο και λευχαιμογόνο δράση της

Valorizing Traditional Greek Wheat Varieties: Phylogenetic Profile and Biochemical Analysis of Their Nutritional Value

Research has highlighted the nutritional benefits of ancient grains, especially emmer (Triticum turgidum ssp. dicoccum) and einkorn (Triticum monococcum), compared to modern varieties of common and durum wheat, focusing on their higher levels of antioxidants and phytochemicals. In this study, grains from old Greek landraces of einkorn, emmer, durum and common wheat were compared to modern wheat cultivars, as well as barley, oats and rye grains, to investigate their unique genetic profile and nutritional properties. Genotyping of Triticum species was performed using SSR markers, which allowed differentiation up to cultivar level. Nutritional factors like the total content of bound and free polyphenols, flavonoids, antioxidant activity and fatty acid profile were assessed. The results showed that emmer and einkorn had the highest total polyphenol, flavonoid and mono-unsaturated fatty acids content, as well as higher antioxidant activity than common and durum wheat. Local landraces of common and durum wheat also exhibited higher values than commercial cultivars. The results of this study demonstrate the high nutritional value of ancient wheat varieties. Many of these cultivars have been put aside by more productive, yet with lower qualitative characteristics, commercial cultivars, underlining the importance of maintaining sustainable agricultural practices to ensure their continued cultivation