Tehran University Medical Journal

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Background: Mutations of ELA2, the gene encoding neutrophil elastase (NE) are known to be associated with cyclic neutropenia (CN) and severe congenital neutropenia (SCN). However, high variability of these mutations has been reported. This study was designed to describe the analysis of the ELA2 gene, clinical manifestations and demographic characteristics in patients with CN and SCN.
Methods: A series of 21 patients with CN or SCN were selected, based on SCINR criteria, from the immunology ward of the Pediatric Medicine Center, Tehran, Iran, from March 2004 to August 2005. The ELA2 gene, isolated from blood samples, was analyzed using RT-PCR and automated capillary sequencing. Informed consent was obtained under the tenets of the Helsinki Declaration and the Ethical Committee of the Tehran University of Medical Sciences.
Results: Kostmann's syndrome and CN was diagnosed in three and 18 patients respectively. Of all the patients, one or two mutations were found in 18 cases (85.7%), including all three patients with SCN and 15 of the patients with CN. Exons two and four had the most mutations (eight and seven cases, respectively). Seven patients had double mutations in two distinct exons. Overall, 16 different mutations were found. At the time of presentation, the mean age of patients was 13.4 ±17.6 months, ranging from one month to seven years. Overall, 61.9% of patients had consanguineous parents. The mean absolute neutrophil count was 830.5 ±419.4 (150-2000)/mm3. On average, each patient had been admitted to the hospital 2.2 ±1.6 times. The neutrophil counts of the SCN patients were significantly higher than those of the CN patients. However, there was no significant difference in the neutrophil counts between patients with mutations and those without mutations. All patients with SCN had two or more infectious complications, although the prevalence of infectious or non-infectious complications did not correlate with ELA2 mutations or the neutropenic disorders.
Conclusion: Mutations in ELA2 appear to play an important role in the phatogenetic mechanisms of CN and SCN. Patients with CN had significantly higher neutrophil counts than SCN patients with CN. Although it possible for the gene encoding neutrophil elastase to have more than one mutation in distinct exons, we found no association between the mutations in ELA2 and their complications in CN and SCN patients.

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Background: Behcet’s disease (BD) is a multisystemic inflammatory disease with unknown origin characterized by recurrent oral aphtous ulcers, genital, ocular and skin lesions. A single point mutation 1691G to A in the factor V gene increases the risk of venous thrombosis. This study designed to determine factor V Leiden mutation in Behcet’s disease, and to find out it's relationship with the clinical manifestations in Khuzestan province, Iran. Methods: One hundred patients with Behcet's Disease (44 males and 56 females) based on international diagnostic criteria and 70 healthy subjects were included in the study.Patients and controls were tested for the presence of factor V Leiden mutation using polymerase chain reaction method. Results: The prevalence of factor V Leiden mutation was significantly higher in BD (10 out of 100, 10%) compared with healthy control subjects (1 out of 70, 1.4%), (p=0.025). Vascular lesions in this study were deep vein thrombosis (DVT) (7%), subcutaneous thrombophlebitis (5%), stroke (1%) and retinal vasculitis (39%). It was found that there was no association between venous thrombosis and the factor V Leiden mutation in Khuzestanian patients. Also, no association between other vascular lesions and the factor V Leiden mutation was recognized. On the other hand there was a significant association between DVT and anterior uveitis (p=0.033). Conclusion: In this study we did not find any association between clinical manifestations in BD patients and factor V Leiden mutation in Khuzestan province, Iran but in BD patients with DVT, factor V Leiden mutation might be a risk factor for the development of anterior uveitis.

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Background: JAK2 is a nonreceptor tyrosine kinase that plays a major role in myeloid disorders. This mutation is characterized by a G to T transverse at nucleotide 1849 in exon 12 of the JAK2 gene, located on the chromosome 9p, leading to a substitution of valine to phenylalanine at amino acid position 617 in the JAK2 protein. In this study we compared the amplification refractory mutation (ARMS) assay and allele- specific (AS- PCR) to evaluate JAK2V617F mutation patients with non-CML myeloproliferative neoplasms (MPNS). Methods: In this experimental study we evaluated JAK2 mutation in 58 patients with a known or suspected diagnosis of a myeloproliferative neoplasm by simple randomized sampling. The mutation was detected by ARMS-PCR and AS-PCR in patients. In order to verify the methods, amplified products from some patients were sequenced. Results: The JAK2 V617F mutation was detected in 86.6%(26/30) of patients with polycythemia vera and 61.5%(8/13) of patients with idiopathic myelofibrosis by ARMS-PCR and AS-PCR. 46.6%(7.15) of essential thrombocythemia patients were positive using ARMS- PCR method while 53%(8.15) of then were positive when AS- PCR were used. The mutation was confirmed by sequencing. Conclusions: The incidence of JAK2 mutation using above PCR methods is similar to previous studies. The different results may depend on the molecular technique used

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Background: Thyroid carcinoma is the most common endocrine malignancy. Medullary thyroid carcinoma (MTC) approximately accounts for 5-10% of all thyroid carcinoma. Nowadays, it is obviously, the mutations in REarranged during transfection (RET) proto-oncogene, especially, mutations in exons 10, 11 and 16 are associated with MTC pathogenesis and occurrence. Thus, early diagnosis of MTC by mutation detection in RET proto-oncogene allows to identify patients who do not have any developed symptoms. The aim of this study was to screening of germline mutations in RET proto-oncogene exons 17 and 18 in MTC patients and their first degree relatives in Iranian population.

Methods: In this cross-sectional study, three hundred eleven participates (190 patients, 121 their relatives) were referred to endocrine research center, Shahid Beheshti University of Medical Science during September 2013 until September 2015. The inclusion criteria were pathological and clinical diagnosis. After whole blood sampling, genomic DNA was extracted from peripheral blood leucocytes using the standard Salting Out/Proteinase K method. Nucleotide change detection in exons 17 and 18 was performed using PCR and direct DNA sequencing methods.

Results: In this study, twenty missense mutations [CGC>TGC, c.2944C>T, p.Arg982Cys (rs17158558)] which included 16 heterozygote and 4 homozygote mutations were found in codon 982 (exon 18). In the present study, 154 G>A (rs2742236) and 4 C>T (rs370072408) nucleotide changes were detected in exons 18 and intron 17 respectively. There was no mutation in exon 17. Conclusion: It seems that because of arginine to cysteine substitutions in RET tyrosine kinase protein structure and its polyphen score (0.955) and SIFT score (0.01) the mutation in codon 982 (exon 18) could be have pathogenic effects. On the other hands, the mentioned mutation frequency was 6.4% among MTC patients, so this mutation of exon 18 could be checked in genetic screening tests of RET proto-oncogene. Although this needs more study.

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