Different deletions identified in Indian type 3 VWD patients.

<p>Different deletions identified in Indian type 3 VWD patients.</p

Prediction software analysis of splice site mutations detected in the present study.

<p>HSF- Human splicing finder.</p><p>Different scoring models used in MAXENTSCAN:</p><p>MAXENT- maximum entropy model.</p><p>MM- first order markov model.</p><p>WMM- weight matrix model.</p

Genetic Heterogeneity in a Large Cohort of Indian Type 3 von Willebrand Disease Patients

<div><p>Background</p><p>Though von Willebrand disease (VWD) is a common coagulation disorder, due to the complexity of the molecular analysis of von Willebrand factor gene (<i>VWF</i>), not many reports are available from this country. Large size of the gene, heterogeneous nature of mutations and presence of a highly homologous pseudogene region are the major impediments in the genetic diagnosis of VWD. The study is aimed at unravelling the molecular pathology in a large series of VWD patients from India using an effective strategy.</p><p>Method</p><p>We evaluated 85 unrelated Indian type 3 VWD families to identify the molecular defects using a combination of techniques i.e. PCR-RFLP, direct DNA sequencing and multiple ligation probe amplification (MLPA).</p><p>Results</p><p>Mutations could be characterized in 77 unrelated index cases (ICs). 59 different mutations i.e. nonsense 20 (33.9%), missense 13 (22%), splice site 4 (6.8%), gene conversions 6 (10.2%), insertions 2 (3.4%), duplication 1 (1.7%), small deletions 10 (17%) and large deletions 3 (5.1%) were identified, of which 34 were novel. Two common mutations i.e. <i>p.R1779*</i> and <i>p.L970del</i> were identified in our population with founder effect. Development of alloantibodies to VWF was seen in two patients, one with nonsense mutation (<i>p.R2434*</i>) and the other had a large deletion spanning exons 16–52.</p><p>Conclusion</p><p>The molecular pathology of a large cohort of Indian VWD patients could be identified using a combination of techniques. A wide heterogeneity was observed in the nature of mutations in Indian VWD patients.</p></div

Different types of mutations identified in type 3 Indian VWD patients.

<p>Different types of mutations identified in type 3 Indian VWD patients.</p

Missense mutations detected in type 3 VWD patients from India.

<p>Missense mutations detected in type 3 VWD patients from India.</p

Splice site mutation identified in Indian type 3 VWD patients.

<p>Splice site mutation identified in Indian type 3 VWD patients.</p

Graph of clinical manifestations seen in severe VWD patients.

<p>Epistaxis (EP), Ecchymosis (EC), On trauma (OT), Gum bleeding (GB), Menorrhagia (MH), Hemarthrosis (HM), On tooth extraction (OX), Malena (MA), Hematuria (HU), Ear bleed (EB),Gastro-intestinal bleeds (GI), On circumcision (OC), Post partum (PP), Bruising (BR), Intra cranial bleed (IC), Post operative (PO), Hematoma at the site of vaccination (HV).</p

Prediction software analysis of novel missense mutations detected in the present study.

<p><b>Polyphen-2</b> - score 0 to1 shows low to high confidence for probability of protein damaging.</p><p><b>SIFT</b>- mutations considered as pathogenic showing score <0.05.</p><p><b>Align GVGD</b>- higher the score higher the probability of protein damaging nature of mutation</p><p><b>MUpro</b> - higher the negativity in SVM score higher the probability of decrease in protein structure stability.</p><p>SVM- Support Vector Machine (SVM score – negative score for novel missense mutations showed the pathogenic probability).</p

Dry weight of iron from the liver and the heart tissues of mice.

<p>* Statistically significant reduction (p<0.001) in dry weight of liver or heart iron of the particular group compared to the group 1</p

Cell derived microparticle levels in women suffering from ≥ 3 recurrent pregnancy loss when compared to controls.

<p>Scatter plots of MP levels of different cell types in different groups of patients along with controls are shown in the 5 graphs. Levels >2 SD from the mean of control women are considered as cut-offs to have increased MP levels. </p