Bioinformatics filtering algorithm.

<p>Shown are bioinformatics analysis tools used and individual and mean variant numbers at each step of filtering with resulting decrease in variant numbers at each decision step.</p

Visualization of NGS alignment and chromatogram from Sanger sequencing confirming the <i>TNNT2 Arg173Trp</i> variant.

<p>The alignment and Sanger sequencing profiles of the TNNT2 R173W variant are shown. A) C>T variant alignment reads of Arg173Trp variant B) (inset) Chromatogram of C>T variant of Arg173Trp variant from Sanger sequencing; arrow depicts the c.517T C>T (chr1∶201,332,477) position.</p

Clinical features of key member of family AD-FDC1 and AD-FDC27.

<p>NYHA - New York Heart Association class; ECG - Electrocardiogram; LVEDD - left ventricular end-diastolic diameter; LVEF - left ventricular ejection fraction; FS - fractional shortening; DOE - Dyspnea on exertion, PND - Paroxysmal nocturnal dyspnea, DCM - Dilated cardiomyopathy, PVC - Premature ventricular contractions, LBBB - Left bundle branch block, CHF - Congestive heart failure, SD - Sudden death, NSVT - Non-sustained ventricular tachycardia, PM - Pace maker, RBBB - Right bundle branch block, AF - Atrial fibrillation, AVB - 1st degree atrio-ventricular block, LAFB - Left anterior fascicular block, SSS - sick sinus syndrome.</p

Venn diagram reflecting variant overlap between and among patients.

<p>A Venn diagram depicts the number of variants after bioinformatics filtering to identify nonsynonymous, nonsense and splice site variants that were located in conserved regions (by ANNOVAR), novel (absent in 1000 Genomes and NHLBI Exome Sequencing Project datasets), and were predicted to be damaging by in silico analyses. The numbers and overlap regions in the Venn diagram show the variants unique to or shared among three individuals (IV:4, IV:7, and IV:14) from family AD-FDC1.</p

Pedigrees of families (A) AD-FDC1 and (B) AD-FDC27.

<p>The family structures of both TNNT2 mutation families are shown. The proband is indicated by an arrow. Males and females are depicted as squares and circles, respectively. Affected individuals are identified by shading. Presence or absence of the Arg173Trp variant confirmed by Sanger sequencing is indicated by plus (‘+’) and minus (‘−’) signs, respectively. Paired numbers beneath individuals represent the numbered haplotypes according to (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0078104#pone-0078104-t004" target="_blank">Table 4</a>); double-numbers represent recombinant haplotypes further detailed in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0078104#pone.0078104.s003" target="_blank">Table S3</a>. Individuals studied by exome sequencing are indicated by present of black underline beneath haplotype numbering.</p

Hap Map project haplotypes and population frequencies of Utah residents with northern and western European ancestry (CEU).

<p>The TNNT2, Arg173Trp variant (Chr1∶201,332,477) was present on separate haplotype, segregating with haplotypes #2 and #1 in families AD-FDC1 and AD-FDC27, respectively.</p

Resultant variants and dbNSFP scores after bioinformatic filtering in AD-FDC1.

<p>Shown are the six genes containing exome-detected variants present in all three subjects tested. The Genebank NM number, chromosome and nucleotide position are shown along with the predicted consequence of each variant. Only the TNNT2 variant segregates with the disease phenotype in AD-FDC1. SIFT, Polyphen2, LRT, and MutationTaster scores derived from dbNSFP are presented. SIFT scores less than 0.05 are predicted to be damaging, otherwise they are predicted to be tolerated. Polyphen2_HDIV_scores range from 0 to 1. Scores in the range of 0.957 to 1 are predicted to be possibly damaging and those in the range of 0.453 to 0.956 are predicted to be benign. Polyphen2_HVAR_scores range from 0 to 1. Scores in the range of 0.909 to1 are predicted to be possibly damaging and scores in the range of 0.447 to 0.908 are predicted to be benign. Lower LRT p-values correspond to predictions that are more damaging. A MutationTaster value close to 1 indicates a high ‘security’ of the prediction.</p

Data_Sheet_1_Evolving trends in epidemiology and natural history of cardiac amyloidosis: 30-year experience from a tertiary referral center for cardiomyopathies.docx

ObjectiveNatural history of cardiac amyloidosis (CA) is poorly understood. We aimed to examine the changing mortality of different types of CA over a 30-year period.Patients and methodsConsecutive patients included in the “Trieste CA Registry” from January 1, 1990 through December 31, 2021 were divided into a historical cohort (diagnosed before 2016) and a contemporary cohort (diagnosed after 2016). Light chain (AL), transthyretin (ATTR) and other forms of CA were defined according to international recommendations. The primary and secondary outcome measures were all-cause mortality and cardiac death, respectively.ResultsWe enrolled 182 patients: 47.3% AL-CA, 44.5% ATTR-CA, 8.2% other etiologies. The number of patients diagnosed with AL and ATTR-CA progressively increased over time, mostly ATTR-CA patients (from 21% before 2016 to 67% after 2016) diagnosed non-invasively. The more consistent increase in event-rate was observed in the long-term (after 50 months) in ATTR-CA compared to the early increase in mortality in AL-CA. In the contemporary cohort, during a median follow up of 16 [4–30] months, ATTR-CA was associated with improved overall and cardiac survival compared to AL-CA. At multivariable analysis, ATTR-CA (HR 0.42, p = 0.03), eGFR (HR 0.98, p = 0.033) and ACE-inhibitor therapy (HR 0.24, p ConclusionIncidence and prevalence rates of ATTR-CA and, to a less extent, of AL-CA have been increasing over time, with significant improvements in 2-year survival of ATTR-CA patients from the contemporary cohort. Reaching an early diagnosis and starting disease-modifying treatments will improve long-term survival in CA.</p