In vivo expression of innate immunity markers in patients with mycobacterium tuberculosis infection

<p>Abstract</p> <p>Background</p> <p>Toll-like receptors (TLRs), Coronin-1 and Sp110 are essential factors for the containment of <it>Mycobacterium tuberculosis </it>infection. The purpose of this study was to investigate the <it>in vivo </it>expression of these molecules at different stages of the infection and uncover possible relationships between these markers and the state of the disease.</p> <p>Methods</p> <p>Twenty-two patients with active tuberculosis, 15 close contacts of subjects with latent disease, 17 close contacts of subjects negative for mycobacterium antigens and 10 healthy, unrelated to patients, subjects were studied. Quantitative mRNA expression of Coronin-1, Sp110, TLRs-1,-2,-4 and -6 was analysed in total blood cells <it>vs </it>an endogenous house-keeping gene.</p> <p>Results</p> <p>The mRNA expression of Coronin-1, Sp110 and TLR-2 was significantly higher in patients with active tuberculosis and subjects with latent disease compared to the uninfected ones. Positive linear correlation for the expression of those factors was only found in the infected populations.</p> <p>Conclusions</p> <p>Our results suggest that the up-regulation of Coronin-1 and Sp110, through a pathway that also includes TLR-2 up-regulation may be involved in the process of tuberculous infection in humans. However, further studies are needed, in order to elucidate whether the selective upregulation of these factors in the infected patients could serve as a specific molecular marker of tuberculosis.</p

ON THE STIMULATION OF FETAL HEMOGLOBIN IN THE ADULT

DURING THE PERINATAL LIFE THERE IS A SWITCH FROM FETAL TO ADULT HEMOGLOBIN PRODUCTION, BUT TRACE AMOUNT OF FETAL GLOBIN EXPRESSION CAN BE DETECTED THROUGHOUT THE ADULT LIFE. KNOWLEDGE OF THE MECHANISMS OF REGULATION OF GLOBIN GENE EXPRESSION IS VERY INTERESTING BECAUSE OF THE ELACIDATION OF THE PHENOMENON AND THE PRACTICAL APPLICATION IN THE VARIOUS HEMOGLOBINOPATHIE IN THE PRESENT WORK WE DESCRIBE EXPERIMENTS (IN VIVO AND IN VITRO) AIMING A) THE UNDERSTANDING OF THE CELLULAR AND MOLECULAR MECHANISMS RESPONSIBLE FOR THE ACTIVATION OF HBF IN THE ADULT AND B) THE CHARACTERIZATION OF FACTORS THAT STIMULATE THE PRODUCTION OF HBFIN THE ADULT. TWO MAJOR PARTS OF EXPERIMENTS ARE DESCRIBED: 1) PHARMACOLOGIC STIMULATION OF HBF IN VIVO IN BABOONS AND MICE USING CYTOTOXIC DRUGS, ERYTHROPOIETIN AND BUTYVATE A 2) EXTRACELLULAR FACTORS IN FETAL CALF SERUM AND NEOPLASTICCELL LINES THAT CAUSE INCREASE OF HBF IN CULTURES.Η ΜΕΤΑΤΡΟΠΗ ΕΜΒΡΥΙΚΗΣ ΑΙΜΟΣΦΑΙΡΙΝΗΣ ΣΕ ΑΙΜΟΣΦΑΙΡΙΝΗ ΕΝΗΛΙΚΟΥ ΣΥΜΒΑΙΝΕΙ ΚΑΤΑ ΤΗΝΠΕΡΙΓΕΝΕΤΙΚΗ ΠΕΡΙΟΔΟ. ΟΜΩΣ, Η ΜΕΤΑΤΡΟΠΗ ΑΥΤΗ ΔΕΝ ΕΙΝΑΙ ΠΛΗΡΗΣ ΚΑΙ ΜΙΚΡΑ ΠΟΣΑ ΕΜΒΡΥΙΚΗΣ ΑΙΜΟΣΦΑΙΡΙΝΗΣ ΕΞΑΚΟΛΟΥΘΟΥΝ ΝΑ ΠΑΡΑΓΟΝΤΑΙ ΚΑΘ'ΟΛΗ ΤΗ ΔΙΑΡΚΕΙΑ ΤΗΕ ΕΝΗΛΙΚΟΥ ΖΩΗΣ. Η ΕΜΒΡΥΙΚΗ ΑΙΜΟΣΦΑΙΡΙΝΗ ΤΟΥ ΕΝΗΛΙΚΟΥ ΜΠΟΡΕΙ ΝΑ ΑΥΞΗΘΕΙ ΣΗΜΑΝΤΙΚΑ ΜΕ ΔΙΑΦΟΡΟΥΣ ΤΡΟΠΟΥΣ. Η ΚΑΤΑΝΟΗΣΗ ΤΟΥ ΜΗΧΑΝΙΣΜΟΥ ΡΥΘΜΙΣΕΩΣ ΤΗΕ ΕΜΒΡΥΙΚΗΣ ΑΙΜΟΣΦΑΙΡΙΝΗΣ ΕΙΣ ΤΟΝ ΕΝΗΛΙΚΟ, ΠΕΡΑ ΑΠΟ ΤΗ ΘΕΩΡΗΤΙΚΗ ΣΥΜΒΟΛΗ ΣΤΗΝ ΚΑΤΑΝΟΗΣΗ ΤΟΥ ΒΙΟΛΟΓΙΚΟΥ ΕΧΕΙ ΙΔΙΑΙΤΕΡΗ ΠΡΑΚΤΙΚΗ ΣΗΜΑΣΙΑ ΓΙΑ ΤΗ ΘΕΡΑΠΕΙΑ ΤΩΝ ΑΙΜΟΣΦΑΙΡΙΝΟΠΑΘΕΙΩΝ. ΣΤΗΝΕΡΓΑΣΙΑ ΑΥΤΗ ΠΕΡΙΓΡΑΦΟΥΜΕ IN VIVO ΚΑΙ IN VITRO ΠΕΙΡΑΜΑΤΑ ΜΕ ΣΚΟΠΟ Α) ΤΗΝ ΚΑΤΑΝΟΗΣΗ ΤΩΝ ΚΥΤΤΑΡΙΚΩΝ ΚΑΙ ΜΟΡΙΑΚΩΝ ΜΗΧΑΝΙΣΜΩΝ ΠΟΥ ΕΙΝΑΙ ΥΠΕΥΘΥΝΟΙ ΓΙΑ ΤΗΝ ΕΝΕΡΓΟΠΟΙΗΣΗ ΤΗΣ ΕΜΒΡΥΙΚΗΣ ΑΙΜΟΣΦΑΙΡΙΝΗΣ ΕΙΣ ΤΟΝ ΕΝΗΛΙΚΟ, ΚΑΙ Β) ΤΗΝ ΑΝΑΚΑΛΥΨΗ ΠΑΡΑΓΟΝΤΩΝ ΟΙ ΟΠΟΙΟΙ ΑΥΞΑΝΟΥΝ ΤΗΝ ΠΑΡΑΓΩΓΗ ΤΗΣ ΕΜΒΡΥΙΚΗΣ ΑΙΜΟΣΦΑΙΡΙΝΗΣ. ΔΥΟ ΚΑΤΗΓΟΡΙΕΣΠΕΙΡΑΜΑΤΩΝ ΠΕΡΙΓΡΑΦΟΝΤΑΙ: 1) ΦΑΡΜΑΚΟΛΟΓΙΚΗ ΕΝΕΡΓΟΠΟΙΗΣΙΣ ΤΗΣ ΕΜΒΡΥΙΚΗΣ ΑΙΜΟΣΦΑΙΡΙΝΗΣ ΜΕ ΧΡΗΣΗ ΚΥΤΤΑΡΟΤΟΞΙΚΩΝ ΦΑΡΜΑΚΩΝ, ΕΡΥΘΡΟΠΟΙΗΤΙΝΗΣ ΚΑΙ ΒΟΥΤΥΡΙΚΟΥ ΟΞΕΟΣ ΣΕ ΠΙΘΗΚΟΥΣ ΚΑΙ ΠΟΝΤΙΚΟΥΣ IN VIVO ΚΑΘΩΣ ΚΑΙ ΣΕ ΚΑΛΛΙΕΡΓΕΙΕΣ ΚΥΤΤΑΡΩΝ ΚΑΙ 2) ΠΕΡΙΒΑΝΤΟΛΟΓΙΚΕΣ ΕΠΙΔΡΑΣΕΙΣ ΣΤΗΝ ΕΚΦΡΑΣΗ ΤΗΣ ΕΜΒΡΥΙΚΗΣ ΑΙΜΟΣΦΑΙΡΙΝΗΣ ΑΠΟ ΠΑΡΑΓΟΝΤΕΣΠΟΥ ΠΕΡΙΕΧΟΝΤΑΙ ΣΤΟΝ ΟΡΟ ΕΜΒΡΥΟΥ ΒΟΥΣ 'Η ΠΑΡΑΓΟΝΤΕΣ ΠΟΥ ΕΚΚΡΙΝΟΝΤΑΙ ΑΠΟ ΝΕΟΠΛΑΣΤΙΚΕΣ ΚΥΤΤΑΡΙΚΕΣ ΣΕΙΡΕΣ

MIRU-VNTR typing of drug-resistant tuberculosis isolates in Greece

The increasing immigration rate in Greece from countries with a high prevalence of Mycobacterium tuberculosis (MTB) and multidrug-resistant tuberculosis (MDR-TB) may have an impact οn the number of MDR-TB cases in Greece. The aim of this study was to genotypically characterize the MTB isolates from patients with pulmonary drug-resistant tuberculosis (DR-TB) in Greece, and to determine whether there is any association between the prevalent genotypes and drug resistance. Fifty-three drug-resistant MTB strains isolated from culture specimens of clinical material from native Greeks and immigrant patients with pulmonary tuberculosis were genotyped using the mycobacterial interspersed repetitive units–variable number of tandem repeats (MIRU-VNTR) method. The phylogenetically distinct groups of isolates identified were: the Beijing (34%), the LAM (11%), the Haarlem (24.5%), the Uganda I (9.4%), the Ural (3.8%), the Delhi/CAS (9.4%) and the Cameroon (3.8%) families. Greek patients were more likely to have monoresistant and polyresistant TB with the most prevalent isolates belonging to the Haarlem family. Among foreign-born patients with MDR-TB, the most prevalent genotypes belonged to the Beijing family. MIRU-VNTR rapidly obtained clinically useful genotyping data, by characterizing clonal MTB heterogeneity in the isolated strains. Our results underline the need for more effective antituberculosis control programs in order to control the expansion of DR-TB in Greece

A case report of recessive restrictive cardiomyopathy caused by a novel mutation in cardiac troponin I (TNNI3)

Abstract Background Restrictive cardiomyopathy is a rare cardiac disease, for which several genes including TNNT2, MYPN, FLNC and TNNI3 have been associated with its familial form. Case presentation Here we describe a female proband with a severely manifested restrictive phenotype leading to heart transplantation at the age of 41, who was found homozygous for the novel TNNI3 mutation: NM_000363.4:c.586G > C, p.(Asp196His). Her parents were third-degree cousins originating from a small village and although they were found heterozygous for the same variant they displayed no symptoms of the disease. Her older sister who was also found heterozygous was asymptomatic. Her twin sister and her brother who were homozygous for the same variant displayed a restrictive and a hypertrophic phenotype, respectively. Their children are all carriers of the mutation and remain asymptomatic until the age of 21. Conclusion These observations point to a recessive mode of inheritance reported for the first time for this combination of gene/disease

Concordance between Three Homologous Recombination Deficiency (HRD) Assays in Patients with High-Grade Epithelial Ovarian Cancer

Our aim was to evaluate the concordance between the Myriad MyChoice and two alternative homologous recombination deficiency (HRD) assays (AmoyDx HRD Focus NGS Panel and OncoScan™) in patients with epithelial ovarian cancer (EOC). Tissue samples from 50 patients with newly diagnosed EOC and known Myriad MyChoice HRD status were included. DNA aliquots from tumor samples, previously evaluated with Myriad MyChoice and centrally reassessed, were distributed to laboratories to assess their HRD status using the two platforms, after being blinded for the Myriad MyChoice CDx HRD status. The primary endpoint was the concordance between Myriad MyChoice and each alternative assay. Tumor samples were evaluated with an AmoyDx® HRD Focus Panel (n = 50) and with OncoScan™ (n = 43). Both platforms provided results for all tumors. Analysis showed that correlation was high for the Myriad MyChoice GI score and AmoyDx® HRD Focus Panel (r = 0.79) or OncoScan™ (r = 0.87) (continuous variable). The overall percent agreement (OPA) between Myriad MyChoice GI status (categorical variable) and each alternative assay was 83.3% (68.6–93.3%) with AmoyDx and 77.5% (61.5–89.2%) with OncoScan™. The OPA in HRD status between Myriad MyChoice and AmoyDx was 88.6% (75.4–96.2). False-positive rates were 31.6% (6/19) for AmoyDx GI status and 31.9% (7/22) for OncoScan™, while false-negative rates were 0% (0/28, AmoyDx) and 11.1% (2/18, OncoScan™) compared with the Myriad MyChoice GI status. While substantial concordance between Myriad MyChoice and alternative assays was demonstrated, prospective validation of the analytical performance and clinical relevance of these assays is warranted

Recommendations for accurate genotyping of SARS-CoV-2 using amplicon-based sequencing of clinical samples

Objectives: Genotyping of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been instrumental in monitoring viral evolution and transmission during the pandemic. The quality of the sequence data obtained from these genotyping efforts depends on several factors, including the quantity/ integrity of the input material, the technology, and laboratory-specific implementation. The current lack of guidelines for SARS-CoV-2 genotyping leads to inclusion of error-containing genome sequences in genomic epidemiology studies. We aimed to establish clear and broadly applicable recommendations for reliable virus genotyping. Methods: We established and used a sequencing data analysis workflow that reliably identifies and removes technical artefacts; such artefacts can result in miscalls when using alternative pipelines to process clinical samples and synthetic viral genomes with an amplicon-based genotyping approach. We evaluated the impact of experimental factors, including viral load and sequencing depth, on correct sequence determination. Results: We found that at least 1000 viral genomes are necessary to confidently detect variants in the SARS-CoV-2 genome at frequencies of &gt;= 10%. The broad applicability of our recommendations was validated in over 200 clinical samples from six independent laboratories. The genotypes we determined for clinical isolates with sufficient quality cluster by sampling location and period. Our analysis also supports the rise in frequencies of 20A.EU1 and 20A.EU2, two recently reported European strains whose dissemination was facilitated by travel during the summer of 2020. Conclusions: We present much-needed recommendations for the reliable determination of SARS-CoV-2 genome sequences and demonstrate their broad applicability in a large cohort of clinical samples. (C) 2021 The Author(s). Published by Elsevier Ltd on behalf of European Society of Clinical Microbiology and Infectious Diseases