Genomic variants in the FTO gene are associated with sporadic amyotrophic lateral sclerosis in Greek patients

Background: Amyotrophic lateral sclerosis (ALS) is a devastating disease whose complex pathology has been associated with a strong genetic component in the context of both familial and sporadic disease. Herein, we adopted a next-generation sequencing approach to Greek patients suffering from sporadic ALS (together with their healthy counterparts) in order to explore further the genetic basis of sporadic ALS (sALS). Results: Whole-genome sequencing analysis of Greek sALS patients revealed a positive association between FTO and TBC1D1 gene variants and sALS. Further, linkage disequilibrium analyses were suggestive of a specific diseaseassociated haplotype for FTO gene variants. Genotyping for these variants was performed in Greek, Sardinian, and Turkish sALS patients. A lack of association between FTO and TBC1D1 variants and sALS in patients of Sardinian and Turkish descent may suggest a founder effect in the Greek population. FTO was found to be highly expressed in motor neurons, while in silico analyses predicted an impact on FTO and TBC1D1 mRNA splicing for the genomic variants in question. Conclusions: To our knowledge, this is the first study to present a possible association between FTO gene variants and the genetic etiology of sALS. In addition, the next-generation sequencing-based genomics approach coupled with the two-step validation strategy described herein has the potential to be applied to other types of human complex genetic disorders in order to identify variants of clinical significance

Molecular and serological study of despersion of borrelia burgdorferi to Macedonia and Thrace

In the present study, a molecular and seroepidemiological examination was performed to investigate the incidence of Lyme disease, or Borreliosis, in Northern Greece. The poor information from the past has not been enough to support whether this infection is present in Greece, and in which frequency, so that the clinical doctors include this infection in the variety of diagnoses. In the present study 2113 ticks were screened for the presence of B. burgdorferi s.l. D.N.A..Τicks were collected from the bodies of freely grazing animals, mainly sheep, goats and cattle, from regions of Northern Greece during the period of April-June and September-December of 2003-2006. The identification of the gender and the stage of development of each tick were determined during observation in a high resolution stereoscope, by the use of dichotomous key, a systematic taxonomy of tick as performed on level genus and species. Two wich amplify partial segment of the gene of rsRNA were performed. The presence of the DNA of the bacteria was also analyzed with the same methodology on 70 samples of renal tissue from different species of rodents that originated from two regions of Northern Greece. Furthermore, the same analysis was investigated in 34 patients (16 men and 18 women) with Bell’s palsy.Στην παρούσα μελέτη έγινε μοριακή και οροεπιδημιολογική διερεύνηση της Lyme μπορρελίωσης στη Βόρειο Ελλάδα. Τα ελάχιστα δεδομένα που υπήρχαν τα προηγούμενα χρόνια δεν ήταν σε θέση να υποστηρίξουν ότι η λοίμωξη αυτή εμφανίζεται στον ελλαδικό χώρο και με ποια συχνότητα, ώστε οι κλινικοί γιατροί να την συμπεριλαμβάνουν στη διαφοροδιάγνωση. Στην παρούσα μελέτη αναζητήθηκε το DNA του μικροοργανισμού σε 2.113 κρότωνες που συλλέχθηκαν από το σώμα ζώων ελευθέρας βοσκής, κυρίως αιγοπρόβατα και βοοειδή, από περιοχές της Β. Ελλάδας κατά το χρονικό διάστημα Απρίλιος-Ιούνιος και Σεπτέμβριος-Δεκέμβριος των ετών 2003-2006. Ο καθορισμός του φύλου και του σταδίου ανάπτυξης κάθε κρότωνα έγινε κατόπιν παρατήρησής του σε στερεοσκόπιο υψηλής ευκρίνειας, ενώ με τη βοήθεια διχοτομικών κλειδών γινόταν παράλληλα και η συστηματική ταξινόμηση αυτού σε επίπεδο γένους και είδους. Όλοι οι συλλεχθέντες κρότωνες εξετάσθηκαν με PCR δοκιμασίες για την ανίχνευση τμήματος δύο διαφορετικών γονιδίων του βακτηρίου, του γονιδίου της βλεφαρίδας (fla gene) και του rsRNA γονιδίου. DNA του βακτηρίου αναζητήθηκε με την ίδια μεθοδολογία και σε δείγματα νεφρικού ιστού από 70 τρωκτικά διαφόρων ειδών που προέρχονταν από δύο περιοχές της Β. Ελλάδας. Αναζητήθηκε επίσης σε 34 ασθενείς (16 άνδρες, 18 γυναίκες) με παράλυση του Bell. Στη συνέχεια ΙgG και IgM αντισώματα έναντι του μικροοργανισμού αναζητήθηκαν σε αντιπροσωπευτικό δείγμα γενικού πληθυσμού ελληνικής εθνικότητας από τους 16 νομούς της Β. Ελλάδας, ηλικίας 0-79 ετών. Συνολικά εξετάσθηκαν 1590 οροί αίματος, που συλλέγησαν κατά την χρονική περίοδο 2003-2005 από ασυμπτωματικά άτομα (750 άνδρες και 840 γυναίκες) που διέμεναν σε αστικές και αγροτικές περιοχές. Αρχικά όλοι οι οροί ελέγχθηκαν για την παρουσία ειδικών IgG και IgΜ αντισωμάτων με δύο ανοσοενζυμικές προκαταρκτικές δοκιμασίες, τη μικροσωματιδιακή δοκιμασία ΜΕΙΑ και τη δοκιμασία ELISA. Κατόπιν οι οροί με θετικό αποτέλεσμα εξετάσθηκαν με την επιβεβαιωτική δοκιμασία ανοσοαποτύπωσης με αντιγόνα την B. burgdorferi sensu stricto και B. afzelii, για τον καθορισμό της ειδικότητας των ανοσοενζυμικών δοκιμασιών. Επιπλέον, οι θετικοί και ύποπτοι οροί με τις ανωτέρω δοκιμασίες εξετάσθηκαν με PCR για την ανίχνευση τμήματος του DNA του βακτηρίου. Η ίδια ορολογική και μοριακή προσέγγιση έγινε και σε 2024 ορούς και 216 ENY ασθενών με συμπτωματολογία ύποπτη για Lyme μπορρελίωση (1045 με νευρολογικά, 894 με ρευματολογικά και 85 με ποικίλα συμπτώματα)

Novel Pathogenic Variants Leading to Sporadic Amyotrophic Lateral Sclerosis in Greek Patients

Amyotrophic lateral sclerosis (ALS) is a rapidly progressive disease that affects motor neurons, leading to paralysis and death usually 3–5 years after the onset of symptoms. The investigation of both sporadic and familial ALS highlighted four main genes that contribute to the pathogenesis of the disease: SOD1, FUS, TARDBP and C9orf72. This study aims to provide a comprehensive investigation of genetic variants found in SOD1, FUS and TARDBP genes in Greek sporadic ALS (sALS) cases. Our sequencing analysis of the coding regions of the abovementioned genes that include the majority of the variants that lead to ALS in 32 sALS patients and 3 healthy relatives revealed 6 variants in SOD1, 19 variants in FUS and 37 variants in TARDBP, of which the SOD1 p.D90A and the FUS c.*356G&gt;A (rs886051940) variants have been previously associated with ALS, while two novel nonsense pathogenic variants were also identified, namely FUS p.R241* and TDP-43 p.Y214*. Our study contributes to the worldwide effort toward clarifying the genetic basis of sALS to better understand the disease’s molecular pathology.</p

Serum Fibroblast Growth Factor 21 Levels in Children and Adolescents with Hashimoto&rsquo;s Thyroiditis before and after l-Thyroxin Medication: A Prospective Study

Backgrounds and Objectives: Fibroblast growth factor 21 (FGF-21) is a complex hormone, sharing common sites of action with thyroid hormones. We investigated the association among FGF-21 levels, resting metabolic rate (RMR), and l-thyroxin (LT4) treatment in children and adolescents with Hashimoto&rsquo;s thyroiditis. Materials and Methods: A total of 60 youngsters with chronic autoimmune thyroiditis (AIT) (30 with subclinical hypothyroidism, 30 with euthyroidism) and 30 age and sex-matched healthy participants (5&ndash;18 years old) were enrolled in the study. Anthropometric, biochemical parameters, and RMR levels were assessed in all participants; serum FGF-21 levels were measured in the control group and the group with subclinical hypothyroidism before and six months after medication with LT4. Results: FGF-21 levels were lower in the treatment group compared with the healthy ones, but this difference was not statistically significant (p &gt; 0.05); despite the increase in FGF-21 levels after six months of LT4 treatment, this difference was not statistically significant (p &gt; 0.05). Free thyroxin (FT4) levels correlated well with FGF-21 levels (r = 0.399, p &lt; 0.01), but further analysis revealed no interaction between these two variables. Both patient groups presented elevated triglyceride (TG) levels compared to controls (p &lt; 0.05). LT4 treatment had no impact on RMR and lipid or liver or glycaemic parameters. An increase in fat mass and fat-free mass were reported, independently of FGF-21 levels. Conclusions: In youngsters with subclinical hypothyroidism due to Hashimoto&rsquo;s thyroiditis, the serum FGF-21 levels are not significantly lower than in healthy individuals and increase after treatment with LT4 without a statistical significance. Further studies with a large number of young patients and severe hypothyroidism are recommended to confirm our results

A Novel Mutation of VPS33B Gene Associated with Incomplete Arthrogryposis-Renal Dysfunction-Cholestasis Phenotype

Arthrogryposis-renal dysfunction-cholestasis (ARC) syndrome is an autosomal recessive disorder caused by mutations of the VPS33B encoding the vacuolar protein sorting 33B (VPS33B), which is involved in the intracellular protein sorting and vesicular trafficking. We report a rare case of ARC syndrome without arthrogryposis caused by a novel mutation of VPS33B. A female patient of Greek origin presented on the 14th day of life with renal tubular acidosis, Fanconi syndrome, nephrogenic diabetes insipidus, and cholestasis with normal gamma-glutamyl transpeptidase, without arthrogryposis and dysmorphic features. She was born to apparently healthy, nonconsanguineous parents. Additional features included dry and scaling skin, generalized hypotonia, hypoplastic corpus callosum, neurodevelopmental delay, failure to thrive, short stature, recurrent febrile episodes with and without infections, and gastrointestinal bleeding. DNA testing revealed that the patient was homozygous for the novel c.1098_1099delTG (p.Glu367Alafs∗17) mutation of exon 14 of VPS33B gene (NM_018668) on chromosome 15q26.1, leading to a nonsense frameshift variant of VPS33B with premature termination of translation. Her parents were heterozygous for the same VPS33B mutation. The prognosis was predictably poor in the context of the intractable polyuria necessitating long-term parenteral fluid administration via indwelling central catheter leading to catheter-related sepsis, to which she eventually succumbed at the age of 7 months. This is the first published VPS33B mutation in an ARC patient of Greek origin. The current case adds to the spectrum of ARC-associated VPS33B mutations and provides evidence supporting the existence of incomplete ARC phenotype. Increased awareness and early genetic testing for ARC are suggested in cases with isolated cholestasis and/or renal tubular dysfunction, even in the absence of arthrogryposis