Extensive genetic screening of Iranian Factor FVII-deficient individuals unraveled several novel mutations and postulated founder effects in some cases.

Background: As the most frequent congenital rare bleeding disorder that transmits in an autosomal recessive manner, factor VII (FVII) deficiency is a serious bleeding complication in populations with high rate of in-marriages. While diagnosis mainly relies on clinical and laboratory phenotypes, plasma FVII antigen and activity levels do not often correlate with symptoms' severity. Objectives: Genetic profiling of the affected individuals potentially improves our biological understanding of this complicated rare disorder. Methods: Conventional polymerase chain reaction-Sanger sequencing and whole-exome sequencing were applied for genetic profiling of F7 gene in 66 symptomatic FVII-deficient individuals from 62 independent pedigrees. Thirty-nine asymptomatic relatives of the patients were also studied. Results: Thirty different F7 pathogenic variations were identified in the studied cases of which 11 have not been reported before. The novel mutations include 5 missenses (c.715G>A, c.794T>C, c.1090C>G, c.1222C>A, c.1265T>C), 3 splicing (c.316+1G>T, c.682-2A>G, c.572-16C>G), 2 nonsenses (c.790delC, c.1248G>A), and 1 frameshift (c.1346delA). A founder effect is proposed for c.790delC that was detected in 8 independent pedigrees who were all from similar geographical regions and ethnic backgrounds. Homozygous c.790delC reduces plasma FVII activity to <1% and causes spontaneous intracranial hemorrhage in early infancy. Conclusion: From the 66 studied symptomatic FVII-deficient individuals, 58 were homozygous carriers of the identified variations. Identification of homozygotes clarifies the potential role of nucleotide variations in reducing FVII activity and their contributions to a certain phenotype. Some of those variations, such as c.1A>G, c.509G>A, c.634C>T, and c.1285G>A have only been previously reported as heterozygous.

Comparison of Endodontic Medicaments on Bond Strength of Fiber Post to Root Dentin Using Resin Cement.

STATEMENT OF THE PROBLEM: Endodontic irrigants and medicaments may affect the bond strength of intracanal posts to root dentin. PURPOSE: The aim of this study was to compare the effect of calcium hydroxide (Ca(OH)2) and 2% chlorhexidine gel (CHX) on bond strength of fiber post cemented with resin cement to root dentin. MATERIALS AND METHOD: This in vitro experimental study was conducted on 36 mandibular premolars. Canals were prepared using the step back technique. After root canal irrigation, the teeth were divided into three groups of 12. Ca(OH)2 paste and CHX gel were used as intracanal medicaments in the first and second groups respectively. No intracanal medicament was used in the third group (control group). Access cavities were then sealed and the teeth were incubated for one week. The root canals were then filled using gutta percha and AH26 sealer and the teeth were incubated for 72 hours. Tooth crowns were then cut at the level of the cementoenamel junction and intracanal posts were placed. The teeth were mounted in auto-polymerizing acrylic resin, and incubated for one week .They were then sectioned into 1.5mm thick slices from their coronal surface using a fully automated cutting machine, and subjected to push-out test until failure. The load at debonding was recorded and data were analyzed using one-way ANOVA, post-hoc test and t-test. The coronal margin of the root was at the level of the surface of acrylic resin in the mold. RESULTS: The mean bond strength was 4.45 MPa in the Ca(OH)2, 2.45 MPa in the CHX and 2.48 MPa in the control group. The difference in this regard was statistically significant among groups (p= 0.04). The Ca(OH)2 group had significant differences with the CHX and control groups (p= 0.03 and p= 0.02, respectively). The difference between the CHX and control groups was not significant (p= 0.974). CONCLUSION: Based on the results, Ca(OH)2 increased the bond strength of fiber post to root dentin but 2% CHX had no effect on bond strength.

A novel D235Y mutation in the GP1BA gene enhances platelet interaction with von Willebrand factor in an Iranian family with platelet-type von Willebrand disease.

Platelet-type von Willebrand disease (PT-VWD) is a rare bleeding disorder with an intrinsic defect in platelets rather than von Willebrand factor (VWF), but has clinical and laboratory features similar to the more common type 2B VWD. The intriguing nature of the pathophysiology and molecular genetics of PT-VWD has created lengthy debate in literature regarding its discrimination from type 2B VWD, and essentially confirming DNA analysis as the gold standard in diagnosis and revealing pathologic mutations. In this report we identify a novel Asp235Tyrmutation in the GP1BA gene of two Iranian patients showing the PT-VWD phenotype who were originally misdiagnosed as type 2B VWD. By structural modelling of the mutant by introducing Tyr235 into the available crystal structure of the glycoprotein (GP)Ibα N-terminal domain, we observed the mutant Tyr235 generates a hydrophobic tip to the extended ß-switch loop of GPIbα. Further modelling of the resulting complex with VWFA1 indicates this could result in an enhanced interface compared to wild-type Asp235. This data provides an update to the present knowledge about this rare disorder, and confirms the necessity of genetic testing for accurate diagnosis, and the importance of studying natural mutations to better understand molecular aspects of GPIbα-VWFA1 interaction.