Journal of Genes and Cells

21-26

Authors: Adel Ghorani-Azam, Seyed Ahmad Mohajeri, Bamdad Riahi-Zanjani, Sona Sepahi, Samaneh Sepahi

Amylase is an enzyme with broad hydrolysis activities that catalyzes the hydrolysis of starch into glucose and other small molecules such as maltose. It is an enzyme of glycoside family with surface layer homology (SLH) domain. SLH, which is known as cell wall binding domain is essential for binding to peptidoglycan of cell membranes. Since designing and producing enzymes with strong catalytic activity is essential in future food industry, investigating the similarities of amylases of different species can be helpful in protein engineering of these enzymes. Homology alignment of bacterial and human amylase sequences shows that both sequences contain 511 amino acid that are highly conserved with several repeated sequences that seems to be necessary for catalysis activity and structural conformation of the protein. In this study, we aimed to characterize the most critical structural and catalytic domains of amylase using bioinformatics tools. In this study, it was shown that SLH and starch binding domain (SBD) are extremely conserved in both human and bacterial amylase. In addition, it was shown that similar domain with several repeated di- or tri-peptide sequences existed in almost all amylases indicating that these residues may have valuable functional, structural and evolutional information. The results of this study may provide new insights into biological aspects of proteins that have not yet been elucidated.

32-35

Authors: Seyed Javad Mowla , Ensieh Poursani, Majid Mehravar, James E. Trosko

Alternative splicing is an important mechanism that regulates gene expression and function in mammals, particularly in primates and human. Large-scale studies estimates a large number of human genes undergo alternative splicing. OCT4, a crucial stem cell marker, is one of the most important stemness markers that is not excluded in this regard. Up until now, it is known that OCT4 can produce several spliced variants in different cell types. In this study, we used RT-PCR and DNA sequencing approaches to verify OCT4 variant expression. Here, we reported a novel OCT4 spliced variant which is named as OCT4B3, for the first time. This variant is very similar to the OCT4B2 transcript, but a 207-bp fragment has been lost from Exon1b. Moreover, we investigated the expression pattern of OCT4B3 in several human cancer cell lines and found its expression in two cell lines of 1321N1 and 5637. Our experiments resulted in discovery of a novel OCT4 variant of OCT4B3 that is expressed in two human 5637 and 1321N1 cell lines.

3-15

Authors: Muhammad Irfan-maqsood, Hojjat Naderi-Meshkin, Asieh Heirani-Tabasi, Monireh Bahrami, Mahdi Mirahmadi, Halimeh Hassanzadeh, Mahmood Raeesolmohaddesin, Nasser Sanjar Moussavi, Hasnain Raza-Shah, Ahmad Reza Bahrami, Hamidreza Bidkhori, Maryam M. Matin

Keratinocytes are the main components of skin epidermis constituting more than 90% of it which are responsible to regulate skin regeneration during external or internal injury. These cells can be found in a heterogenic form containing proliferative, terminally differentiated and transit amplifying (TA) cells located in basal layer, outer keratinized layer and intermediate layers, respectively when isolated from an adult skin. Efforts are going on to characterize keratinocytes precisely as in comparison with widely used stem cells, mesenchymal stem cells (MSCs) and their application in cutaneous wound healing as a translational approach in regenerative medicine. In this work, we have applied a comprehensive approach to identify and characterize keratinocytes which are a valuable tool in wound healing. Keratinocytes have successfully been isolated from adult skin and the ratio of proliferative, terminally differentiated and TA cells based on the expression of α6-integrin and CD71 was studied using flow cytometry after 7, 14 and 21 days of their culture in vitro. RT and qRT-PCR was applied to study the change in genetic expression and relativity of cytokeratin markers with the passage of time. PopUp culturing and population doubling time was performed to study the proliferative potential of heterogenic population of cells. Their colonogenicity and wound healing potential were also studied to explain their healing behaviour during the time of injuries. 47% keratinocytes after 7 days, 62% after 14days and 93% keratinocytes after 21 days of culturing. Expression of CD71 was also observed in cells as, 5% after 7 days, 22% after 14 days and 66% of keratinocytes after 21 days were positive for CD71 expression. Gradual increased expression of genetic markers, K10, K14, INV and P63 was observed using qRT-PCR. Studying comparative data, it can be concluded that keratinocytes after 14 days of culturing are better to use for clinical purposes. This comprehensive protocol could be a valuable addition in cutaneous biology helping researchers to identify keratinocytes and to use them for their pre-clinical studies.

8-10

Authors: Naveed Akhtar , Kausar Saeed, Naseem Rafiq, Nasreen Khan

Dengue is one of the leading cause of death worldwide. The first outbreak of dengeu hemorrhagic fever was reported in 1994 in Pakistan and after that several outbreaks of the dengue were reoprted in Pakistan and in all these outbreaks the dengue serotype 2 (DENV2) was confirmed, which ensures that dengue serotype 2 (DENV2) is prevelant in Pakistan.

16-20

Authors: Afeefa Kiran Ch

Dengue fever is an epidemic killing millions of every year around the world especially in regions with moderate temperature primarily due to propagation of the vector, Aedes aegypti. Unknown genomic basis is the biggest challenge to compete with this challenge. A little information is available about the genotypic characterization of the virus infecting the population. Based on the genotype, dengue viruses are classified into four types named as DENV-1, DENV-2, DENV-3, DENV-4. All these serotypes are composed of several proteins know as NS proteins which are further categories into NS1, NS2A, NS2B, NS3, NS4A,NS4B, NS5 based on their functions. In this manuscript, detailed of serotypes classification, their components proteins, their structure and functions are discussed briefly which will be helpful to understand the basic of dengue viral genome.