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کاربرد نوع شرط:
- جایگاه : پژوهشی
- مجله: Iranian Journal of Basic Medical Sciences
- نوع مقاله: Journal Article
- کلمات کلیدی: Mutation,Promoter,CAPZA3,Failed fertilization,ICSI,PLCζ
- چکیده:
- چکیده انگلیسی: Objective(s): Phospholipase C ζ (PLCζ) is considered as a nominee for sperm associated oocyte activating factors and is located back-to-back with CAPZA3, an actin-capping protein controlling actin polymerization during spermiogenesis. They contain a common bidirectional promoter. The objective of this study was to identify individuals with parallel low expression of PLCζ and CAPZA3 mRNA, in hope of detecting genetic defects in this bidirectional promoter. Materials and Methods: Semen samples were collected from 24 fertile and 59 infertile individuals with total failed, low and high fertilization rate post intra-cytoplasmic sperm injection (ICSI), as well as globozoospermic individuals.Expression of PLCζ and CAPZA3 were assessed by Real time PCR. In addition, PLCζ was assessed by Western blot. Results: Significant correlations between PLCζ with CAPZA3 and also between these two genes with fertilization were observed. Individuals with low fertilization presented significantly lower expression of these two genes. Low expression of PLCζ was also verified by Western analysis. Sequence analysis of bidirectional promoter of these two genes in an individual with parallel low expression of both PLCζ and CAPZA3, revealed a mutation within the CAPZA3 predicted promoter, known as human regulatory factor X4 which is a testis-specific dimeric DNA-binding protein. In the opposite stand, in the same location, the mutation appears to be outside but in the vicinity of PLCζ, in a binding region predicate by Genomatix. Conclusion: Parallel assessment of CAPZA3 with PLCζ at mRNA level in individuals with inability to induce oocyte activation may help researcher to identify genetic defects associated with failed fertilization.
- انتشار مقاله: 27-12-1394
- نویسندگان: Soudabeh Javadian-Elyaderani,Kamran Ghaedi,Marziyeh Tavalaee,Farzaneh Rabiee,Mohammad Reza Deemeh,Mohammad Hossein Nasr-Esfahani
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Basic Medical Sciences
- نوع مقاله: Journal Article
- کلمات کلیدی: Alginate,Autologous,chondrocytes,Collagen Type II,Proteoglycan,Transplantation
- چکیده:
- چکیده انگلیسی: Objective(s) Articular cartilage tissue defects cannot be repaired by the proliferation of resident chondrocytes. Autologous chondrocyte transplantation (ACT) is a relatively new therapeutic approach to cover full thickness articular cartilage defects by in vitro grown chondrocytes from the joint of a patient. Therefore, we investigated the redifferentiation capability of human chondrocytes maintained in alginate culture. Materials and Methods The cartilage specimens obtained from 50 patients who underwent total knee and hip operations at the teaching hospital of Isfahan University of Medical Sciences, Isfahan Iran. Isolated primary chondrocytes were first grown in monolayer cultures for 1 to 6 passages (each passage lasting about 3 days). At each passage, monolayer cells seeded in alginate culture and investigated morphologically and immuno-cytologically for expression of cartilage-specific markers (collagen type II and cartilage-specific proteoglycans). Results The chondrocytes from monolayer passages P1 to P4 introduced in alginate cultures regained a chondrocyte phenotype. Cells were interconnected by typical gap junctions and after few days, they produced a cartilage-specific extracellular matrix (collagen type II and cartilage-specific proteoglycans). In contrast, cells from monolayer passages P5 and P6 did not redifferentiate to chondrocytes in the alginate cultures. Conclusion Chondrocyte culture was established for the first time in Iran. The alginate culture conditions promote the redifferentiation of dedifferentiated chondrocytes that have still a chondrogenic potential. This procedure opens up a promising approach to produce sufficient numbers of differentiated chondrocytes for ACT. Indeed, in some patients the harvested cells were used immediately and successfully for transplantation.
- انتشار مقاله: 14-07-1394
- نویسندگان: Ebrahim Esfandiary,Mehdi Shakibaei,Noshin Amirpour,Mehrafarin Fesharaki,Mohammad Hossein Nasr-esfahani,Fariba Moulavi,Farahnaz Moulavi,Khalilollah Nazem,Masoud Hatef Dehghani,Constanze Csaki,Shahnaz Razavi
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Biotechnology
- نوع مقاله: Journal Article
- کلمات کلیدی: Promyelocytic Leukemia,Pluripotent stem cells,retinoic acid
- چکیده:
- چکیده انگلیسی: Background: Promyelocytic leukemia protein (PML) is a tumor suppressor protein that is involved in myeloid cell differentiation in response to retinoic acid (RA). In addition, RA acts as a natural morphogen in neural development. Objectives: This study aimed to examine PML gene expression in different stages of in vitro neural differentiation of NT2 cells, and to investigate the possible role of PML in pluripotency and/or neural development. Materials and Methods: RA was used as a neural inducer for in vitro neural differentiation of NT2 cells. During this process PML mRNA and protein levels were assessed by quantitative real time RT-PCR (QRT-PCR) and Immunoblotting, respectively. Furthermore bisulfite sequencing PCR (BSP) was used to assess PML promoter methylation in NT2 cells and NT2 derived neuronal precursor cells (NT2.NPCs). Results: QRT-PCR results showed that, PML had maximum expression with significant differences in NT2 derived neuronal precursor cells relative to NT2 cells and NT2 derived neural cells (NT2.NCs). Numerous isoforms of PML with different intensities appeared in immunoblots of pluripotent NT2 cells, NT2.NPCs, and NT2.NCs. Furthermore, the methylation of the PML promoter in NT2.NCs was 2.6 percent lower than NT2 cell. Conclusions: The observed differences in PML expression in different cellular stages possibly could be attributed to the fact that PML in each developmental state might be involved in different cell signaling machinery and different functions. The appearance of different PML isoforms with more intensity in neural progenitor cells; may suggest apossible role for this protein in neural development.
- انتشار مقاله: 28-05-1394
- نویسندگان: Khadijeh Karbalaie,Sadeq Vallian,Liana Lachinani,Somayeh Tanhaei,Hossein Baharvand,Mohammad Hossein Nasr-Esfahani
- مشاهده
- جایگاه : پژوهشی
- مجله: Iranian Journal of Biotechnology
- نوع مقاله: Journal Article
- کلمات کلیدی: Inverse PCR,phiC31 Integrase,Pseudo attP Site
- چکیده:
- چکیده انگلیسی: Background: PhiC31 integrase is a DNA site-specific recombinase integrates DNA into the chromosomes between the two sites of attB and attP. Several pseudo attPs have been identified in mammalian genomes with critical features for long-term expression of transgene. In this manuscript, we report a novel intrinsic pseudo attP site named CHOL1 in the Chinese hamster genome implementing an inverse Polymerase Chain Reaction (IPCR). Objectives:Identification of pseudo attP site(s) of Tenecteplase cDNA integration in the genome of stable transformed CHO cell line. Materials and Methods: First, genome was extracted from a stable transformed CHO cell line expressing Tenecteplase. By creating of minicircle DNA in the last step, sequencing was performed. Results: We obtained one band. BLAST analysis of the respective sequence of inverse PCR band identified a pseudo attP site. Conclusions: Data demonstrated that the phiC31 integrase provides a suitable insertion site in the genome to express the gene of interest.
- انتشار مقاله: 25-10-1391
- نویسندگان: Mohammad Hadi Sekhavati,Kianoush Dormiani,Kamran Ghaedi,Yahya Khazaie,Morteza Hosseini,Mojtaba Tahmoorespur,Mohammad Reza Nassiri,Mahboubeh Foruzanfar,Mohammad Hossein Nasr Esfahani
- مشاهده
- جایگاه : پژوهشی
- مجله: International Journal of Pediatrics
- نوع مقاله: Journal Article
- کلمات کلیدی: Fibroblast,Peroxisome biogenesis disorder,PEX7,PTS2,RCDP
- چکیده:
- چکیده انگلیسی: Background
Rhizomelic chondrodysplasia punctata (RCDP) type 1 is among of the rare autosomal recessive peroxisome biogenesis disorders caused by mutations in the PEX7 gene. RCDP patients with the classic form of RCDP1 do not live more than 10- year.
Materials and Methods
In the present study, a two-year-old girl with skeletal abnormalities and dysmorphic facial appearance is reported to be suffered from RCDP. The patient's parents were second cousins and healthy and there was no similar case in the parents’ family. PEX7 gene was sequenced in the patient and her parents.
Results
A homozygous mutation, G257A, was identified PEX7 in the genome of patient while the parents were compound heterozygous.
Conclusion
Taken together, clinical presentation and peroxisome profile of the patient suggested a missense mutation led to formation of a pathogenic PEX7, responsible for incidence of RCDP.- انتشار مقاله: 19-06-1396
- نویسندگان: Marziyeh Alamatsaz,Kamran Ghaedi,Motahare-Sadat Hashemi,Yousef Shafeghati,Mohammad Faghihi,Mohammad Hossein Nasr-Esfahani
- مشاهده
- جایگاه : پژوهشی
- مجله: Asian Pacific Journal of Cancer Prevention
- نوع مقاله: Journal Article
- کلمات کلیدی: children,Parents,risk factors,Acute Lymphoblastic Leukemia
- چکیده:
- چکیده انگلیسی: Background: Acute lymphoblastic leukemia (ALL) is the most common malignancy in children. Different
environmental factors might be effective in the occurrence of this malignancy during childhood. The aim of this study
was to find environmental risk factors in childhood ALL in Hamadan, Iran. Methods: This case-control study was
done in 2015-2018 on 125 children younger than 15 years of age suffering from ALL. Patients were matched with
130 controls with respect to age, gender, and residence location. The identification of risk factors for ALL was sought
based on the comparison of studied variables between case and control individuals. Results: A statistically significant
increased risk for ALL was found with regard to type of delivery (OR: 0.43, 95% CI: 0.20 - 0.92, p˂0.02), childcare
(OR: 4.58, 95% CI: 0.95 - 22.20, p˂0.04), birth weight (OR: 1.44, 95% CI: 1.53 - 2.21, p˂0.006), father’s education
level (OR: 2.67, 95% CI: 1.10 - 6.45, p˂0.02), and father’s job (OR: 0.2 95% CI: 0.08 - 0.51, p˂0.001). Also observed
were increased odds for ALL regarding male gender, mother’s high education level, mother’s freelance job, and medium
or high family income. No association with ALL incidence was observed for age, gender, breastfeeding, mother’s age
at pregnancy, malignancy in first- or second-degree relatives, or mother’s use of hair dye during pregnancy (p> 0.05).
Conclusion: This study showed that father’s education level, father’s job, delivery type, birth weight, and childcare
can play a role in the incidence of childhood ALL.- انتشار مقاله: 07-03-1398
- نویسندگان: Hassan Rafieemehr,Fereshteh Calhor,Hossein Esfahani,Somayeh Ghorbani Gholiabad
- مشاهده