Effects of the SNP rs536714306 on <i>GPR126</i> signal transduction in HEK293T cells.

<p>(A) HEK293T cells were transfected with empty vector, wild-type GPR126, or mutant GPR126 containing SNP rs536714306 for 24 h, and expression of GPR126 and β-Actin was assessed by western blotting. Asterisk represents a non-specific band. (B) HEK293T cells were transfected with empty vector, wild-type GPR126, or mutant GPR126 for 48 h. Following a 1 h incubation with 3 μg/mL type IV collagen, cAMP expression was assessed by ELISA. Data represent the average and standard deviation of three independent experiments. *<i>p</i> <0.05. EV, empty vector; WT, wild-type GPR126; mut, mutant GPR126.</p

List of 10 filtered variants.

<p>List of 10 filtered variants.</p

Primer list for real-time PCR.

<p>Primer list for real-time PCR.</p

Expression of GPR126 during the cytodifferentiation of HPDL cells.

<p>HPDL cells were transfected with empty vector and incubated with mineralization medium for 4, 6, 9, and 15 days and the expression of BSP and GPR126 was assessed by real-time PCR. Data represent the average and standard deviation of four independent experiments. *<i>p</i> <0.05.</p

GWAS findings.

<p>GWAS findings.</p

Clinical characteristic of individual subjects in the GWAS.

<p>Clinical characteristic of individual subjects in the GWAS.</p

A Development of Nucleic Chromatin Measurements as a New Prognostic Marker for Severe Chronic Heart Failure

<div><p>Background</p><p>Accurate prediction of both mortality and morbidity is of significant importance, but it is challenging in patients with severe heart failure. It is especially difficult to detect the optimal time for implanting mechanical circulatory support devices in such patients. We aimed to analyze the morphometric ultrastructure of nuclear chromatin in cardiomyocytes by developing an original clinical histopathological method. Using this method, we developed a biomarker to predict poor outcome in patients with dilated cardiomyopathy (DCM).</p><p>Methods and Results</p><p>As a part of their diagnostic evaluation, 171 patients underwent endomyocardial biopsy (EMB). Of these, 63 patients diagnosed with DCM were included in this study. We used electron microscopic imaging of cardiomyocyte nuclei and an automated image analysis software program to assess whether it was possible to detect discontinuity of the nuclear periphery. Twelve months after EMB, all patients with a discontinuous nuclear periphery (Group A, n = 11) died from heart failure or underwent left ventricular assist device (VAD) implantation. In contrast, in patients with a continuous nuclear periphery (Group N, n = 52) only 7 patients (13%) underwent VAD implantation and there were no deaths (p<0.01). We then evaluated chromatin particle density (Nuc-CS) and chromatin thickness in the nuclear periphery (Per-CS) in Group N patients; these new parameters were able to identify patients with poor prognosis.</p><p>Conclusions</p><p>We developed novel morphometric methods based on cardiomyocyte nuclear chromatin that may provide pivotal information for early prediction of poor prognosis in patients with DCM.</p></div

Distribution of Chromatin Score Values (Nuc-CS and Per-CS) and Cardiac Events.

<p>(A) Relationship between Nuc-CS and cardiac events. A white circle represents a patient who did not experience a cardiac event and a dark circle represents a patient who experienced a cardiac event. (B) Relationship between Per-CS and cardiac events. A white circle indicates a patient who did not experience a cardiac event and a dark circle represents a patient who experienced a cardiac event. (C) Receiver operating characteristic (ROC) curve analysis for Nuc-CS in predicting ventricular assist device (VAD) implantation 1 year after endomyocardial biopsy (EMB). (D) ROC curve analysis for Per-CS predicting VAD implantation 1 year after EMB. (E) ROC curve analysis for BNP (black), LVEDD (light blue), LVEF (dark blue), and %Fib (gray) in predicting VAD implantation 1 year after EMB. AUC, area under the curve; BNP, brain natriuretic peptide; LVEDD, left ventricular end-diastolic diameter; LVEF, left ventricular ejection fraction; %Fib, area of fibrosis within a specimen.</p

Algorithm for DCM Therapy Based on Myocardial Biopsy Parameters.

<p>The first step is morphometric analysis of cardiomyocyte nuclei. When chromatin in the nuclear periphery is discontinuous and there is a homogeneous aggregation of particles in the nucleoplasm, the patient is classified as Group A, which is associated with poor prognosis. When a patient does not have these characteristics, a second step, which involves additional quantitative analysis, is added. Based on the chromatin scores Nuc-CS and Per-CS, a treatment plan can be determined.</p

Clinical Outcomes of Groups A and N.

<p>Clinical Outcomes of Groups A and N.</p