ABSTRACT
BACKGROUND: Hypertrophic cardiomyopathy is a common genetic cardiac disease. Prevention and early diagnosis of this disease are very important. Because of the large number of causative genes and the high rate of mutations involved in the pathogenesis of this disease, traditional methods of early diagnosis are ineffective. METHODS: We developed a custom AmpliSeq panel for NGS sequencing of the coding sequences of ACTC1, MYBPC3, MYH7, MYL2, MYL3, TNNI3, TNNT2, TPM1, and CASQ2. A genetic analysis of student cohorts (with and without cardiomyopathy risk in their medical histories) and patients with cardiomyopathies was performed. For the statistical and bioinformatics analysis, Polyphen2, SIFT, SnpSift and PLINK software were used. To select genetic markers in the patients with cardiomyopathy and in the students of the high risk group, four additive models were applied. RESULTS: Our AmpliSeq custom panel allowed us to efficiently explore targeted sequences. Based on the score analysis, we detected three substitutions in the MYBPC3 and CASQ2 genes and six combinations between loci in the MYBPC3, MYH7 and CASQ2 genes that were responsible for cardiomyopathy risk in our cohorts. We also detected substitutions in the TNNT2 gene that can be considered as protective against cardiomyopathy. CONCLUSION: We used NGS with AmpliSeq libraries and Ion PGM sequencing to develop improved predictive information for patients at risk of cardiomyopathy.
Subject(s)
Calsequestrin/genetics , Cardiac Myosins/genetics , Cardiomyopathy, Hypertrophic/diagnosis , Carrier Proteins/genetics , Chest Pain/diagnosis , Myosin Heavy Chains/genetics , Software , Troponin T/genetics , Adolescent , Adult , Aged , Calsequestrin/blood , Cardiac Myosins/blood , Cardiomyopathy, Hypertrophic/blood , Cardiomyopathy, Hypertrophic/genetics , Carrier Proteins/blood , Chest Pain/blood , Chest Pain/genetics , Cohort Studies , Early Diagnosis , Female , Gene Expression , Genetic Markers , High-Throughput Nucleotide Sequencing , Humans , Male , Middle Aged , Models, Genetic , Myosin Heavy Chains/blood , Open Reading Frames , Risk , Troponin T/bloodABSTRACT
BACKGROUND: Overt ophthalmopathy is presumed to be uncommon in patients with Hashimoto's thyroiditis compared to Graves' disease, where significant eye changes are found in approximately 40% of patients. On the other hand, when observing, more subtle eye changes, particularly upper eyelid retraction (UER) and mild inflammatory signs, may be common in patients with Hashimoto's thyroiditis. METHODS: We have determined the prevalence and characteristics of eye signs in recently diagnosed patients with Hashimoto's thyroiditis studied prospectively since 2004 till date in Sydney (Australia). We measured serum orbital antibodies in 20 of the patients in enzyme-linked immunosorbent assay. RESULTS: The overall prevalence of eye signs in patients with Hashimoto's thyroiditis was 34%, of whom about a quarter had chronic UER, determined as a margin-reflex distance of >5 mm, as the main sign. There was no correlation between eye signs and cigarette smoking. Overall, there was only a modest correlation between eye signs and positive antibody tests, and 40% of patients with no eye signs at the time of study were antibody positive. CONCLUSION: Eye changes, in particular UER, are common in patients with Hashimoto's thyroiditis. Since thyroid stimulating hormone-receptor antibodies are not usually associated with Hashimoto's thyroiditis, autoimmune mediated damage of the levator palpebrae superioris (eyelid) muscle cannot be due to these antibodies. Although eyelid abnormalities may be a minor problem for most patients, for some there are major cosmetic implications requiring surgical management.
Subject(s)
Eye Diseases/complications , Hashimoto Disease/complications , Adolescent , Adult , Aged , Aged, 80 and over , Autoantibodies/blood , Calsequestrin/blood , Collagen Type XIII/blood , Enzyme-Linked Immunosorbent Assay , Eye/immunology , Eye Diseases/blood , Eye Diseases/immunology , Eyelids/immunology , Female , Hashimoto Disease/blood , Hashimoto Disease/immunology , Humans , Male , Middle Aged , Statistics, NonparametricABSTRACT
BACKGROUND: In heart failure, alterations in the expression of proteins relevant to calcium homeostasis are involved in depressed contractility and diminished relaxation. However the regulation of genes expression is only partially known. The aim was to assess expression of calcium regulatory proteins in left ventricle (LV) myocardium characterised by a preserved global function in mitral valve stenosis (MVS) model but increased neurohumoral/cytokine (N/C) activation. METHODS AND RESULTS: Plasma N/C activation was evaluated in MVS-patients (n = 27), where expression of calcium regulatory proteins (L-type channel, sarcoplasmic reticulum Ca2+-ATPase type2 - SERCA2, Na+/Ca2+ exchanger -NCX, calsequestrin, phospholamban) in LV myocardium was assessed (Western Blot) in comparison with non-failing hearts (NFH). Out of all proteins assessed in MVS, only SERCA2 and NCX expression revealed highly variable changes between subjects, with significant reduction of SERCA2 (15%) level compared to NFH. Moreover, SERCA2 was negatively correlated with BNP (univariate/regression analysis r = -0.63, P = 0.005/r2 = 0.74, P <0.001, respectively), whereas NCX was positively correlated only with noradrenaline (univariate/stepwise analysis r = 0.59 P = 0.002/r2 = 0.59; P = 0.003). CONCLUSIONS: In MVS-patients LV becomes remodelled, although its global function is preserved. It seems that apart from alterations in LV load and wall stress, also such neurohumoral factors as BNP/noradrenaline may influence the Ca2+ handling proteins expression.
