Detection of Chlamydia pneumoniae (Chlamydophila pneumoniae) DNA in atherosclerotic plaques and its molecular analysis in northern Greece

Objectives: C. pneumoniae responsible for respiratory tract infections has also been assocciated with chronic diseases such as atherosclerosis.The aim of the present study is the detection of C. pneumoniae DNA in various atherosclerotic arteries by a sensitive and specific PCR. In order to investigate whether there is a relation between a specific type and atherosclerosis, genotyping was performed. Methods: The study group consisted of 122 atherosclerotic plaques from patients (mean age 68.4, range 50-89 years old, 95 males and 25 females) with severe atherosclerosis. C. pneumoniae DNA was detected in atherosclerotic plaques by nested «Touchdown» PCR. A second PCR targeting the ygeD-urk intergenic region was performed and PCR products were sequenced.Results: 12.3% of the specimens were positive for C. pneumoniae. Detection rates in specimens of carotid, abdominal, and femoral arteries were 12%, 15.6%, and 10%, respectively. (p = NS). 14 strains were found to have 100% homology with J138, AR39 and TW-183, while one strain had a 23 bp invertible region and revealed 100% homology with the CWL029.Conclusion: Overall, 15/122 (12.3%) atherosclerotic specimens from patients were positive for C. pneumoniae. The strains detected belong to two different types designated as genotype I and II. Genotype I was the prevalent and only one strain had the reverse orientation of the 23bp region in northern Greece

Associations between total, free and bioavailable 25-hydroxyvitamin D forms with adiponectin and irisin in maternal-neonatal pairs at birth from Greece

Background: Apart from the well-established skeletal effects, vitamin D has been explored as a secretagogue influencing various adipokines, including adiponectin and irisin. Recent evidence suggests that specific forms of 25-Hydroxyvitamin D (25(OHD), such as free and bioavailable 25(OH)D, may provide more accurate measurements of vitamin D status. The relationship between vitamin D status and serum irisin and adiponectin concentrations remains largely unexplored, particularly during pregnancy. Methods: We analyzed data from 67 healthy maternal-neonatal pairs from Northern Greece at birth. Biochemical and hormonal tests were conducted on each maternal-neonatal pair. The vitamin D forms were estimated using validated mathematical models. Subsequently, regression analyses were conducted to determine the association between the vitamin D forms and adipokine levels. Results: Bioavailable maternal 25(OH)D was inversely associated with neonatal irisin concentrations [β=-73.46 (-140.573 to -6.341), p=0.034]. No other associations were observed between maternal vitamin D status and neonatal adipokine concentrations. Conclusion: In conclusion, maternal bioavailable vitamin D concentrations are inversely associated with neonatal serum irisin concentrations, warranting further studies to evaluate the underlying mechanisms for this finding

Salivary Markers for Oral Cancer Detection

Oral cancer refers to all malignancies that arise in the oral cavity, lips and pharynx, with 90% of all oral cancers being oral squamous cell carcinoma. Despite the recent treatment advances, oral cancer is reported as having one of the highest mortality ratios amongst other malignancies and this can much be attributed to the late diagnosis of the disease. Saliva has long been tested as a valuable tool for drug monitoring and the diagnosis systemic diseases among which oral cancer. The new emerging technologies in molecular biology have enabled the discovery of new molecular markers (DNA, RNA and protein markers) for oral cancer diagnosis and surveillance which are discussed in the current review

STRUCTURAL AND FUNCTIONAL ANALYSIS OF MOUSE AND HUMAN GLUTAMATE DEMYDROGENASE GENES. MOLECULAR ANALYSIS OF GLUD1 GENE IN CERTAIN HUMAN NEUROLOGICAL DISEASES

THE AIM OF THIS STUDY WAS THE STRUCTURAL AND FUNCTIONAL ANALYSIS OF GLUTAMATEDEHYDROGENASE GENE FAMILY BOTH IN MOUSE AND HUMAN AND ALSO THE ELUCIDATION OF THE MOLECULAR BASIS OF FAMILIAR AND SPORADIC FORMS OF CERTAIN HUMAN DEGENERATIVE DISORDERS ASSOCIATED WITH A DEFICIENCY OF GLUTAMATE DEHYDROGENASE. IN THIS CONTEST WE ISOLATED AND CHARACTERAZED A FULL LENGTH MOUSE BRAIN GLUDCDNA. MAPPING STUDIES BY USING A WELL-CHARACTERIZED PANEL OF SOMATIC CELL HYBRIDS REVEALED THE EXISTENCE OF TWO MOUSE GLUTAMATE DEHYDROGENASE GENES LOCATED ON CHROMOSONE 14 (GLUD) AND 7 (GLUD-2). PERLIMINARY ANALYSISOF THESE LOCI REVEALED THAT THE GLUD LOWER IS THE FUNCTIONAL GENE, WHILE GLUD-2 RAPRESENTS A FRUNCAFED PSEUDOGENE. THE STUCTURE OF THE PROMOTOR OF GLUD GENE HAS THE CHARACTERISTICS OF THE HOUSEKEE PING PROMOTERS. NORTHERN BLOT ANALYSIS REVEALED THE PRESENCE OF TWO GLUD MRNAS SPECIES DIFFERING BOTH IN SIZE AND THE RELATIVE ABUNDANCES. PRELIMINARY SOUTHERN BLOT ANALYSIS SHOWED A COMPLEX HYBRIDIZATION PATTERN IN AGGREEMENT WITH THE PRESENCE OF A SMALL GLUDGENE FAMILY INHUMANS. THE DECREASED GLUD MRNA LEVELS IN PATIENTS WITH OLIVOPONTOCEREBELLAR ATROPHY ARE NOT DUE TO MUTATIONS IN GLUD1 LOCUR.ΣΚΟΠΟΣ ΑΥΤΗΣ ΤΗΣ ΔΙΑΤΡΙΒΗΣ ΗΤΑΝ Η ΜΕΛΕΤΗ ΤΗΣ ΔΟΜΗΣ ΚΑΙ ΛΕΙΤΟΥΡΓΙΑΣ ΤΩΝ ΓΟΝΙΔΙΩΝ ΤΗΣ ΓΛΟΥΤΑΜΙΚΗΣ ΑΦΥΔΡΟΓΟΝΑΣΗΣ (GLUD)ΣΕ ΔΥΟ ΠΟΛΥ ΟΜΟΛΟΓΑ ΣΥΣΤΗΜΑΤΑ ΤΟΥ ΠΟΝΤΙΚΟΥ ΚΑΙ ΤΟΥ ΑΝΘΡΩΠΟΥ ΚΑΙ Η ΔΙΕΡΕΥΝΗΣΗ ΤΗΣ ΜΟΡΙΑΚΗΣ ΒΑΣΗΣ ΟΡΙΣΜΕΝΩΝ ΟΙΚΟΓΕΝΩΝ ΚΑΙ ΣΠΟΡΑΔΙΚΩΝ ΝΕΥΡΟΕΚΦΥΛΙΣΤΙΚΩΝ ΤΟΥ ΑΝΘΡΩΠΟΥ ΠΟΥ ΣΥΝΔΕΟΝΤΑΙ ΜΕ ΑΝΕΠΑΡΚΕΙΑ ΤΟΥ ΕΝΖΥΜΟΥ ΚΑΙ ΑΝΩΜΑΛΟ ΜΕΤΑΒΟΛΙΣΜΟΤΟΥ ΓΛΟΥΤΑΜΙΚΟΥ. ΣΤΑ ΠΛΑΙΣΙΑ ΑΥΤΗΣ ΤΗΣ ΠΡΟΣΠΑΘΕΙΑΣ ΕΓΙΝΕ Η ΑΠΟΜΟΝΩΣΗ ΚΑΙ Ο ΠΡΟΣΔΙΟΡΙΣΜΟΣ ΤΗΣ ΝΟΥΚΛΕΟΤΙΔΙΚΗΣ ΑΛΛΗΛΟΥΧΙΑΣ ΜΟΡΙΩΝ GLUD CDUA. ΠΕΙΡΑΜΑΤΑ ΧΡΩΜΟΣΩΜΑΤΙΚΗΣ ΧΑΡΤΟΓΡΑΦΗΣΗΣ ΕΔΕΙΞΑΝ ΤΗΝ ΠΑΡΟΥΣΙΑ ΓΕΝΕΤΙΚΩΝ ΤΟΠΩΝ ΓΙΑ ΤΗ GLUD ΣΤΟ ΓΟΝΙΔΙΩΜΑ ΤΟΥ ΠΟΝΤΙΚΟΥ ΠΟΥ ΕΔΡΑΖΟΝΤΑΙ ΣΤΟ ΧΡΩΜΟΣΩΜΑ 14 (GLUD) ΚΑΙ 7(GLUD-2). ΠΡΟΣΔΙΟΡΙΣΤΗΚΕ ΤΟ ΣΗΜΕΙΟ ΕΝΑΡΞΗΣΤΗΣ ΜΕΤΑΓΡΑΦΗΣ ΚΑΙ Η 5 ΠΛΕΥΡΙΚΗ ΠΕΡΙΟΧΗ ΤΟΥ ΛΕΙΤΟΥΡΓΙΚΟΥ ΓΟΝΙΔΙΟΥ GLUD. Ο ΠΟΙΟΤΙΚΟΣ ΚΑΙ ΠΟΣΟΤΙΚΟΣ ΕΛΕΓΧΟΣ ΤΩΝ ΕΠΙΠΕΔΩΝ GLUDMRNA ΑΠΟΚΑΛΥΨΕ ΤΗΝ ΠΑΡΟΥΣΙΑ ΔΥΟ ΜΕΤΑΓΡΑΦΩΝ ΠΟΥ ΔΙΑΦΕΡΟΥΝ ΤΟΣΟ ΣΤΟ ΜΕΓΕΘΟΣ ΟΣΟ ΚΑΙ ΣΤΗΝ ΣΧΕΤΙΚΗ ΑΦΘΟΝΙΑ ΤΟΥΣ. ΣΤΟΝ ΑΝΘΡΩΠΟ ΠΡΟΚΑΤΑΡΤΙΚΗ ΑΝΑΛΥΣΗ ΚΑΤΑ SOUTHERN ΑΠΟΚΑΛΥΨΕ ΕΝΑ ΠΟΛΥΠΛΟΚΟΠΡΟΤΥΠΟ ΥΒΡΙΔΟΠΟΙΗΣΗΣ ΥΠΟΔΕΙΚΛΥΟΝΤΑΣ ΕΤΣΙ ΤΗΝ ΠΑΡΟΥΣΙΑ ΜΙΑΣ ΜΙΚΡΗΣ ΓΟΝΙΔΙΑΚΗΣ ΟΙΚΟΓΕΝΕΙΑΣ ΓΙΑ ΤΗΝ GLUD. ΤΑ ΜΕΙΩΜΕΝΑ ΕΠΙΠΕΔΑ GLUD MRNAΣΕ ΑΣΘΕΝΕΙΣ ΜΕ ΕΛΑΙΟΓΕΦΥΡΟΠΑΡΕΓΚΕΦΑΛΙΔΙΚΗ ΑΤΡΟΦΙΑ ΔΕΝ ΦΑΙΝΕΤΑΙ ΝΑ ΟΦΕΙΛΟΝΤΑΙ ΣΕ ΚΑΠΟΙΑ ΕΝΔΕΧΟΜΕΝΗ ΜΟΡΙΑΚΗ ΒΛΑΒΗ ΤΟΥ ΓΟΝΙΔΙΟΥ GLUD1

Therapeutic Potentials of Reducing Liver Fat in Non-Alcoholic Fatty Liver Disease: Close Association with Type 2 Diabetes

Nonalcoholic fatty liver disease (NAFLD), the most widespread chronic liver disease worldwide, confers a significant burden on health systems and leads to increased mortality and morbidity through several extrahepatic complications. NAFLD comprises a broad spectrum of liver-related disorders, including steatosis, cirrhosis, and hepatocellular carcinoma. It affects almost 30% of adults in the general population and up to 70% of people with type 2 diabetes (T2DM), sharing common pathogenetic pathways with the latter. In addition, NAFLD is closely related to obesity, which acts in synergy with other predisposing conditions, including alcohol consumption, provoking progressive and insidious liver damage. Among the most potent risk factors for accelerating the progression of NAFLD to fibrosis or cirrhosis, diabetes stands out. Despite the rapid rise in NAFLD rates, identifying the optimal treatment remains a challenge. Interestingly, NAFLD amelioration or remission appears to be associated with a lower risk of T2DM, indicating that liver-centric therapies could reduce the risk of developing T2DM and vice versa. Consequently, assessing NAFLD requires a multidisciplinary approach to identify and manage this multisystemic clinical entity early. With the continuously emerging new evidence, innovative therapeutic strategies are being developed for the treatment of NAFLD, prioritizing a combination of lifestyle changes and glucose-lowering medications. Based on recent evidence, this review scrutinizes all practical and sustainable interventions to achieve a resolution of NAFLD through a multimodal approach

Dabrafenib, a selective BRAFV600E inhibitor, is not able to reverse the established CpG island methylator phenotype in RKO colorectal cell line

Background: Colorectal cancer is considered as one of the most common death causes among cancer types in the developed countries. Treatment of colorectal cancer is a challenge due to its molecular heterogeneity originating from genetic mutations and epigenetic events such as hypermethylation of human genome. Methylation in the promoter of genes shows specific patterns, which define the molecular pathogenesis and prognosis of cancer. Therefore, reversal of DNA methylation constitutes a potential therapeutic target. Coexistence of B-RAF V600E mutation with hypermethylation in the promoter of specific genes and chromosomal instability characterize the serrated pathway of carcinogenesis in colorectal cancer and has been associated with poor prognosis. The purpose of this study was to investigate if inhibition of BRAF V600E mutation by the selective inhibitor dabrafenib in the RKO cell line has any effect on the methylation phenotype of the Weisenberger’s CIMP panel genes, CACNA1G, NEUROG1, RUNX3, IGF2 and SOCS1.Materials and methods: RKO cell line cells were cultured under various conditions of Dabrafenib concentrations, time of treatment, cell passage and culture medium provision. Cells from every condition were counted and the subsequently extracted DNA was modified using sodium bisulfate. The characterization of the methylation phenotype was performed by MS-PCR analysis. Modified genomic DNA from Caco2 was used as a control.Results: Dabrafenib treatment resulted in a 50% inhibition of cell growth rate, independent of the concentration used and has no effect on the methylation status of the genes tested under all conditions.Conclusions: Inhibition of the B-RAFV600E by dabrafenib was not able to alter (reverse) the methylation phenotype of CACNA1G, NEUROG1, RUNX3, IGF2 and SOCS1 genes in the RKO cell line