Identification of the First Iranian Family with “γArg275Cys” Mutation (Fibrinogen Tokyo II)

Background: Inherited fibrinogen deficiencies areclassified into two categories: quantitative, including afibrinogenemia and hypofibrinogenemia and qualitative, including dysfibrinogenemia. Any mutation in fibrinogen genes accounts for one of these disorders.Case report: This article reports an Iranian family with dysfibrinogenemia without any clinical signs accidentally diagnosed by routine coagulation tests with slightly elevated PT and APTT a few years ago. Fordeterminationof disease causing genetic aberration in fibrinogen genes, DNA sequencing of three hot spots of these genes (i.e. exon 2 of FGA,exon 2 of FGB and exon 8 of FGG)was performed. Analysis of sequencing results revealed a heterozygous missense mutation c.901 C>T (Arg275Cys) in exon 8 of FGG in mother and children.No mutationwas detected in father’s sample.Fibrinogen with this mutation is known as Tokyo II.Conclusion: γArg275Cys is a heterozygous mutation that impairs the function of fibrinogen andhas been solely reported in dysfibrinogenemic patients. Clinical findings in this family (no history of bleeding and thrombosis) were compatible with molecular results, because fibrinogen Tokyo II does not have a thrombotic or hemorrhagic nature and lack of clinical signs in this family is not unexpected.

Bleeding Episodes Among Patients with Congenital Fibrinogen Disorders, a Study On 12 New Iranian Patients

Background: Congenital fibrinogen disorders (CFDs) comprise about 10% of rare bleeding disorders (RBDs). CFDs are divided into two groups of quantitative (afibrinogenemia and hypofibrinogenemia) with autosomal recessive inheritance pattern, and qualitative (dysfibrinogenemia, hypodysfibrogenemia) disorders, mainly with autosomal dominant inheritance pattern. Sistan and Baluchestan Province in Iran, with its high rate of consanguineous marriages, has a high incidence of RBDs including CFD. In the current study, we report clinical manifestations of patients with CFDs.Methods: Twelve new Iranian patients from Sistan and Baluchestan Province with different types of CFDs were selected for this study. Diagnosis of CFDs was based on clinical features and familial history followed by laboratory assessment by routine and specific coagulation tests including prothrombin time (PT) and activated partial time tests (APTT), as well as FI activity assay by Clauss method.Results: Out of 12 patients, 3(25%) had afibrinogenemia, 7(58.3%) had hypofibrinogenemia while 2(16/7%) were suspected of having dysfibrinogenemia. Although umbilical cord bleeding (UCB) 9(75%) was the most common clinical presentation among the study population, this feature was not observed among patients with dysfibrinogenemia. Hematoma (100%) was the most common presentation of patients with dysfibrinogenemia.  Conclusion: Results of this study revealed that some clinical presentations are the diagnostic features of CFDs and can be used for precise and in-time diagnosis CFDs in conjunction with family history and laboratory findings.Keywords: Fibrinogen Deficiency; Congenital Afibrinogenemia; Blood Coagulation Disorder; Afibrinogenemi

Coagulation Factor XIII-A A614T gene Variation is Suggestive of Founder Effect in Iranian Patients with Severe Congenital Factor XIII Deficiency

Background: Factor XIII (FXIII) is a heterotetramer consisting of two subunits, FXIII-A and FXIII-B. Several common gene variations were observed in the FXIII-A gene with an obvious ethnic difference. This study assessed the prevalence of A614T as a common FXIII-A gene variation among Iranian patients with FXIII deficiency (FXIIID). Materials and Methods: This study was conducted on eighty Iranian unrelated individuals with FXIIID. Genotype analysis for FXIII-A A614T gene variation was performed for all individuals. Results: Molecular analysis of these Iranian populations revealed that all studied patients were homozygous for the T allele at codon 204 of the FXIII-A1 subunit. Conclusion: Present of T allele at codon 204 of FXIII-A1 subunit among all study population can be suggestive of founder effect. &nbsp

Thrombin Activatable Fibrinolysis inhibitor Thr 325 Ile polymorphism in fetuses with factor XIII deficient family history and Intracranial hemorrhage

Background: Factor XIII Deficiency (FXIIID) is an inherited rare bleeding disorder with some life threatening clinical manifestation including Intracranial Haemorrhage (ICH). Among all polymorphisms found in FXIIID, Thrombin Activatable Fibrinolysis Inhibitor (TAFI) Thr325Ile gene polymorphism increases probability of ICH about 20 fold in patients with FXIII .So, in this study we aimed to evaluate TAFI Thr 325 Ile polymorphism in Chorionic villus samples (CVS) of fetuses with positive family history of FXIIID and ICH.Materials and Methods: This study was performed on chorionic villus of pregnant mothers ´ with positive history of FXIIID accompanied with ICH in first-degree relatives of their fetus. All parents of the fetuses were completed consent form for doing Prenatal diagnosis (PND). Chorionic villus DNA was extracted from each sample using the DNA extraction kit and PCR-RFLP was performed for TAFI Thr 325Ile polymorphism in Exon 4 of FXIII A gene.Results: All of 8 fetuses had positive family history of FXIIID. Seven out of eight fetuses (87.5%) had a family member with CNS bleeding due to FXIIID. Four fetuses had history of death due to FXIIID. There were 5 case (62.5%) that were homozygote for TAFI Thr 325 Ile, one (12.5%) was heterozygote and two (25%) were non mutant. Conclusion: Detection of TAFI Thr 325 Ile polymorphism by PND program in fetuses with positive family history of ICH is seems necessary and it will help to fill many gaps in preventing life threatening features of FXIIID in newborn at the time of delivery by prophilaxy receiving and precautionary measures

Expression of Long Non-Coding RNA H19 in Acute Lymphoblastic Leukemia

Objective: Long non-coding RNA (lncRNA) H19 has essential roles in growth, migration, invasion, and metastasis ofmost cancers. H19 dysregulation is present in a large number of solid tumors and leukemia. However, the expressionlevel of H19 in acute lymphoblastic leukemia (ALL) has not been elucidated yet. The current study aimed to exploreH19 expression in ALL patients and cell lines. Materials and Methods: This experimental study was conducted in bone marrow (BM) samples collected from 25patients with newly diagnosed ALL. In addition, we cultured the RPMI-8402, Jurkat, Ramos, and Daudi cell linesand assessed the effects of internal (hypoxia) and external (chemotherapy medications L-asparaginase [ASP] andvincristine [VCR]) factors on h19 expression. The expressions of H19, P53, c-Myc, HIF-1α and β-actin were performedusing quantitative real-time polymerase chain reaction (qRT-PCR) method. Results: There was significantly increased H19 expression in the B-cell ALL (B-ALL, P<0.05), T-cell ALL (T-ALL,P<0.01) patients and the cell lines. This upregulation was governed by the P53, HIF-1α, and c-Myc transcriptionfactors. We observed that increased c-Myc expression induced H19 expression; however, P53 adversely affected H19expression. In addition, the results indicated that chemotherapy changed the gene expression pattern. There was aconsiderable decrease in H19 expression after exposure to chemotherapy medications; nonetheless, hypoxia inducedH19 expression through P53 downregulation. Conclusion: Our findings suggest that H19 may have an important role in pathogenesis in ALL and may act as apromising and potential therapeutic target

Serum Vascular Endothelial Growth Factor (VEGF) levels in patients with alpha thalassemia

 Background: Alpha-thalassemia syndrome includes a group of hereditary anemia in which expression of alpha globin chains is decreased or absent. Impaired RBC in patients with thalassemia causes vessel involvement and endothelial cell vessel disturbance. Vascular Endothelial Growth Factor (VEGF) is the most important regulator for endothelial cell proliferation. So, the aim of this study is to compare the serum VEGF levels in patients with alpha thalassemia and normal control group.Materials and Methods: This case-control study was conducted on 17 patients with alpha thalassemia and 40 healthy people. Serum VEGF levels were measured by enzyme-linked immune sorbent assay (ELISA) kit. Then statistical analysis of results were performed using SPSS 16, value of P &lt;0.05 was considered statistically significant.Results: Mean serum VEGF levels in case and control groups were 2294.19±1552.39 and 598.09±988.17pg/ml, respectively. Serum VEGF levels were higher in patients with alpha thalassemia (P &lt;0.01). There was no significant correlation between serum VEGF levels and Hemoglobin. (P= 0.73).Conclusion: Our study revealed that patients with alpha thalassemia have elevated levels of serum VEGF than normal control group. Further studies with larger sample size are recommended to confirm these observations

Upregulation of miR-210 promotes differentiation of mesenchymal stem cells (MSCs) into osteoblasts

Numerous studies indicated that microRNAs are critical in the regulation of cellular differentiation, by controlling the expression of underlying genes. The aim of this study was to investigate the effect of miR-210 upregulation on differentiation of human umbilical cord blood (HUCB)-derived mesenchymal stem cells (MSCs) into osteoblasts. MSCs were isolated from HUCB and confirmed by their adipogenic/osteogenic differentiation and flow cytometric analysis of surface markers. Pre-miR-210 was amplified from human DNA, digested and ligated with plenti-III-mir-green fluorescent protein (GFP) vector, and cloned in STBL4 bacteria. After confirmation with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), the plenti-III-GFP segment bearing pre-miR-210 was transfected into MSCs by electroporation. Two control vectors, pmaxGFP and Scramble, were transfected separately into MSCs. The expression of miR-210 and genes related to osteoblast differentiation, i.e., runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and osteocalcin gene, in the three groups of transfected MSCs was analyzed 0, 7, 14, and 21 days of transfection by quantitative reverse transcription PCR (qRT-PCR). Overexpression of miR-210 was observed in MSCs transfected with miR-210-bearing plasmid, and this was significantly different compared to Scramble group (p < 0.05). Significantly increased expression of Runx2 (at day 7 and 14), ALP and osteocalcin genes (at all time points for both genes) was observed in MSCs with miR-210-bearing plasmid compared to controls. Overall, the overexpression of miR-210 in MSCs led to MSC differentiation into osteoblasts, most probably by upregulating the Runx2, ALP, and osteocalcin genes at different stages of cell differentiation. Our study confirms the potential of miRNAs in developing novel therapeutic strategies that could target regulatory mechanisms of cellular differentiation in various disease states

The Effect of miR-940 Up-regultion on HbF and Erythroid Markers Expression in k-562 Cell Line

Abstract Background: Fetal hemoglobin (α2γ2) is the main oxygen transport protein in the human fetus. Fetal hemoglobin is nearly completely replaced by hemoglobin A, except in a few thalassemia cases and sickle cell anemia. Several studies have indicated that expression of γ-globin might be regulated post-transcriptionally. Small non-coding RNA called microRNAs which target mRNA can lead to translated repression or mRNA decay. The aim of this study is to investigate the effect of miR-940 up-regultion on γ-chain gene expression and erythroid markers in k562 cell line. Materials and Methods: In this experimental study, k562 cells were cultured in RPMI1640. Then pre miR-940 was transfected by electroporation method in k562 cell line. In 3, 7 and 14 days, RNA was extracted and cDNA synthesized in selected days. Up-regulation of miR-940 was confirmed by miRNA Quantitative real time PCR and then the expression γ of chains and GATA-1 was investigated by QRT-PCR. Finally, erythroid markers were checked by flow cytometry. Results: In 3, 7 and 14 days after transfection, the GATA-1 and γ-chain expression were increased in comparison with untransfected cells. Also, the expression of erythroid markers was increased. Conclusion: The data show that up-regulation of miR-940 has a role in the increase of γ-chain gene expression in k-562 cell line. We suggest that miR-940 may be a significant potent therapeutic target for increasing Hb F level. Patients with sickle cell anemia and β-thalassemia are suitable candidate for treatment in this way

Correlation between Gastro Intestinal Failure and mortality rate of ICU admitted patients in Valiasr hospital of Arak city in 2011.

Background: According to insufficient clinical studies concerning association between Gastro Intestinal Failure Score (GIF Score) and mortality rate of ICU admitted patients, the aim of this study was to determine predicting value of GIF Score in mortality rate of ICU admitted patients. Materials and methods: In this cross-sectional study, 357 patients who were &ge;18y/o and were admitted in ICU during 72 hrs were enrolled in the study. For all patients, informed consent form, demographic data sheet and physical examination were completed and intra-abdominal pressure, GIF Score of first week and mortality rate of first week and 28th day were recorded. All collected data was analyzed by SPSS 16 software. Results: In this study mortality rate of 7th and 28th day were 11.5% and 29.7%, respectively. Incidence of high intra-abdominal pressure (IAH) and abdominal compartment syndrome (ACS) were 34.7% and 6.2%. The mean GIF Score during first week was 1.346(SD 0.935). All of these variables were significantly correlated with mortality of 7th and 28th day. Also in logistic regression model, IAH, ACS, GIF score were predicting variables for mortality of 7th and 28th day in ICU admitted patients. Conclusion: According to the results GIF score was significantly correlated with mortality rate of 7th and 28th day in ICU admitted patients. It seems further multi central studies are essentia

Promoter methylation status and expression levels of rassf1a gene in different phases of acute lymphoblastic leukemia (All)

Background: Although the precise pathogenesis of acute lymphoblastic leukemia (ALL) remains unclear, studying gene-regulating mechanisms during ALL pathogeneses may shed light on the underlying mechanisms driving malignant behavior. There is some evidence showing the promoter hypermethylation and silencing of RASSF1A tumor suppressor gene in ALL cells; however, there is a lack of evidence for whether the gene indeed alters during different phases of ALL or in response to therapy. Thus, the current study aimed to clarify this issue using groups of adult ALL patients who have been scarcely investigated regarding expression levels and promoter methylation status. Materials and Methods: In this case/control study, the expression levels and methylation status of the gene promoter was evaluated using quantitative real-time PCR and methylation-specific PCR (MSP), respectively in adults with ALL. The study included peripheral blood of patients with newly diagnosed ALL (n=10), complete remission (CR) (n=10), or relapse (n=10), and 10 control samples from healthy individuals. Results: MSP results revealed an unmethylated status for almost all patients and control samples, except a case with relapsing ALL, which showed a hemimethylated pattern. RASSF1A also showed no difference in terms of gene expression in the patients compared with the control group (p>0.05). Conclusion: The results revealed an up-regulation of RASSF1A tumor suppressor in adult ALL patients experiencing CR, suggesting this to be a marker of therapy response. However, further investigations using more sensitive methylation detecting tools with larger sample sizes may better clarify the involvement of the promoter methylation of RASSF1A in these patients