Avicenna Journal of Medical Biotechnology
Volume:6 Issue: 2, Apr-Jun 2014

A network can be loosely defined as a structure linking together individual and organizational actors with shared goals or values، though often not a shared geography. A large body of literature highlights the important interaction between knowledge and networks. Interest in the impact of networking on knowledge translation and exchange، diffusion of innovations، knowledge management، and organizational outcomes is also increasing 1. There has been a growing interest in research networks and its implications on the creation of new knowledge. For example، there seems to be a consensus that those «scientists who collaborate with each other are more productive، oftentimes producing ''better'' science، than are individual investigators. An open science platform can empowers researchers in their daily work and where everybody has equal opportunity to seek، share and generate knowledge. A value network can be defined as a network of relationships، which creates both tangible and intangible value through a complicated dynamic exchange between individuals، groups and organisations 2. The partnership for research and innovation in the health system funding opportunity recognizes the need to create networks of health researchers and clinical practitioners that can generate solutions to improve sustainable quality and value for money in the health system. The partnership for health research and innovation in the health system will support research and innovation. It seems that the concept of research networking in developing countries with several limitations such as research budgets should be engraved in the minds rather than papers.

Fragile Histidine Triad protein (FHIT), as a known tumor suppressor protein, has been proposed to play crucial role in inhibiting p53 degradation by MDM2. Studies have confirmed FHIT interaction with p53 or MDM2, although functional interacting domains of FHIT with MDM2 and/or p53 are not completely defined. Thus, through determining the significant structural interacting domains of FHIT, information with regard to MDM2 and p53 would be provided. As there were no previous studies evaluating the interaction of optimized important parts of target molecules, docking study was employed. Truncated structures of FHIT were screened to reveal critical sections engaging in FHIT interaction. HEX program was used in order to study the interaction of target structures. Given the total energy, FHIT structures (β5-7, α1) and (α1) of FHIT were showed to be better candidates in comparison with other structures in interaction with optimized MDM2 part. Furthermore, FHIT structures (β4-7, α1) and (β5-7, α1) were considered to be better than other structures in interaction with optimized p53 part. FHIT truncates which interact with MDM2 optimized part exhibited lower energy levels than FHIT truncates which interact with p53 optimized part. Our results can be useful for designing new inhibitors of this protein complex interaction which would result in tumor repression.

Asthma is caused by the combination of different factors. Current concepts of asthma pathogenesis emphasize on gene-environment interactions. Mega-genome scanning projects revealed that different Single Nucleotide Polymorphisms (SNPs) are related to asthma susceptibility. rs7216389-T is one of them that is related to childhood asthma and its effect on childhood asthma severity has been proved in different nations, however no study has been performed in Eastern Mediterranean and Middle East countries yet. To perform population genetic studies, a rapid and high-throughput screening method is necessary. High-resolution melting analysis is a rapid, powerful and accurate method, which is suitable for this type of studies. Therefore, it has been decided to develop a high-resolution melting method for rs7216389 locus genotyping in Iranian asthmatic children. In the current study, a high-resolution melting analysis method based on SYBR Green-I was developed to check the frequency of rs7216389-T mutation in Iranian asthmatic children for the first time. Second and third classes of intercalating dyes are commonly used for high-resolution melting method. However, in this study, SYBR Green-I was used for rs7216389 locus genotyping for the first time. Our results for 60 samples showed that SYBR Green-I has good efficacy for rs7216389 locus genotyping through high-resolution melting method in comparison with PCR-RFLP and sequencing. Comparison of our results based on HRM analysis with PCR-RFLP showed that our developed method is rapid, accurate, high-throughput and economic to study the rs7216389 locus in asthmatic children and it is applicable for other similar population genetic studies.

Tubulin protein being the fundamental unit of microtubules is actively involved in cell division thus making them a potential anti-cancer drug target. In spite of many reported drugs against tubulin, few of them have started developing resistance in human β-tubulin due to amino acid substitutions. In this study we generated three mutants (F270V, A364T and Q292E) using Modeller9v10 which were targeted with compounds from higher and lower plants along with marine isolates using iGEMDOCK2.0 to identify their residual interactions. The mutant F270V does not bring in any increase in the binding affinity in comparison with the taxol-wild type due to their conservative substitutions. However, it increases the volume of the active site. A364T mutant brings a better binding among few of the marine and higher plants isolates due to the substitution of the non-reactive methyl group with the polar residue. But this leads to reduced active site volume. Finally the mutant Q292E from epothilone binding site brings a remarkable change in drug binding in the mutants in comparison with the wild type due to the substitution of uncharged residue with the charged one. But as such there was no change in the volume of the active site observed in them. Lower plants extracts were reported to exhibit better interactions with the taxol and epothilone binding sites. Whereas marine and higher plants isolates shows significant interactions only in the wild type instead of the mutants. In addition to this, the residual substitutions were also found to alter the conformations of the active sites in mutants.

The well documented source for adult multipotent stem cells is Spermatogonial Stem Cells (SSCs). They are the foundation of spermatogenesis in the testis throughout adult life by balancing self-renewal and differentiation. The aim of this study was to assess the effect of percoll density gradient and differential plating on enrichment of undifferentiated type A spermatogonia in dissociated cellular suspension of goat testes. Additionally, we evaluated the separated fractions of the gradients in percoll and samples in differential plating at different times for cell number, viability and purification rate of goat SSCs in culture. Testicular cells were successfully isolated from one month old goat testis using two-step enzymatic digestion and followed by two purification protocols, differential plating with different times of culture (3, 4, 5, and 6 hr) and discontinuous percoll density with different gradients (20, 28, 30, and 32%). The difference of percentage of undifferentiated SSCs (PGP9.5 positive) in each method was compared using ANOVA and comparison between the highest percentage of corresponding value between two methods was carried out by t-test using Sigma Stat (ver. 3.5). The highest PGP9.5 (94.6±0.4) and the lowest c-Kit positive (25.1±0.7) in Percoll method was significantly (p≤0.001) achieved in 32% percoll gradient. While the corresponding rates in differential plating method for the highest PGP9.5 positive cells (81.3±1.1) and lowest c-Kit (17.1±1.4) was achieved after 5 hr culturing (p<0.001). The enrichment of undifferentiated type A spermatogonia using Percoll was more efficient than differential plating method (p<0.001). Percoll density gradient and differential plating were efficient and fast methods for enrichment of type A spermatogonial stem cells from goat testes.

Transforming Growth Factor-beta (TGF-β) activation appears to be crucial for tissue injury in Diabetic Nephropathy (DN). Fibromodulin, the small leucine-rich proteoglycan, has been proposed to be the potent TGF-β modulator. In this study, the therapeutic effects of fibromodulin in the kidneys of streptozotocin (STZ)-induced diabetic rats were investigated. Diabetic rats received intraperitoneal (IP) injections of recombinant adenovirus expression vectors (RAd5) containing fibromodulin (RAd-FMOD) and were killed after 10 weeks. Proteins were isolated from the rat kidney and separated using two-dimensional gel electrophoresis. The differentially expressed proteins were analyzed using Matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Ten spots were identified using MALDI-TOF-MS. The identified proteins were primarily responsible for cell metabolism, cytoskeleton formation, and oxidative stress. RAd-FMOD treatment markedly attenuated the albuminuria in diabetic rats. Taken together, these results provide a valuable clue in exploring the mechanism underlying the therapeutic effects of fibromodulin in diabetic nephropathy suggesting that it can be a potential agent in the treatment of this disease.

Numerous in vitro reports suggest that Low Level Laser Therapy (LLLT) affects cellular processes by biostimulation, however most of them emphasize on using visible light lasers which have low penetration. The aim of this study was to determine the effect of infrared laser light (which is more useful in clinic because of its higher penetration) on secretion of Fibroblast Growth Factor (FGF), Platelet Derived Growth Factor (PDGF) and Vascular Endothelial Growth Factor (VEGF), as important growth factors in wound healing. Fibroblasts were extracted from the skin of 7 diabetic and 7 nondiabetic mice and cultured. Cell cultures of experimental group were irradiated with single dose of LLLT (energy density of 1 J/cm2) using an 810 nm continuous wave laser and the control group was not irradiated. Secretion of growth factors by skin fibroblasts were quantified through real time polymerase chain reaction. Diabetic irradiated group showed significant increase in FGF (p=0.017) expression, although PDGF increased and VEGF decreased in both diabetic and nondiabetic irradiated groups, but these variations were not statistically significant. These results suggest that LLLT may play an important role in wound healing by stimulating the fibroblasts.

Somites are transient developmental structures and the derivatives of paraxial mesoderm which have an important role in organization of segmented pattern of vertebrate embryos 1. In vitro simulation of developmental mechanisms of vertebrates، especially mammals، could help us to find out the intrinsic molecular events involved in the development of organs and differentiation of various kinds of cells like CNS neurons. It is almost impossible to isolate developmental structures such as somite and notochord in human to study their functions in vitro. Even in laboratory mammals like mice، somites are too small to isolate and meticulous work should be done for separating them. Thus، the sole way is to separate such structures in chicken 2. In such situation، chick embryos are the best choice for somite isolation and in vitro simulation of developmental process of neurons. This simulation helps us to find out whether human embryonic cells respond to the molecular signals sent by somite، thereby exploring a new way for regeneration of damaged neural tissues. Previous studies 3 have shown that co-culture of somites derived from chick embryos of stages 9-12 of Hamburger and Hamilton 4 caused an increase in TUJ1/HOXB4 double positive group among human neural progenitors. Also it has been shown by Sagha et al 5 that somites maintain their neural induction ability in mouse embryonic stem cells. They have shown that somites thicken the adjacent part of neural tube، thereby affect the proliferation of neural tube precursors 6. So far، there is no confirmation on expression of secretory factors by somite، whether they are able to co-express several neurogenic factors like noggin، chordin، follistatin، cerberus and FGF8 in vitro. The neurogenic activity of noggin 7، chordin 8، follistatin 9، cerberus 10 and FGF8 11 has already been established. Thus، this study investigated the co-expression of the mentioned factors by somites at mRNA level.

Angiogenesis، which is required for embryonic development and many physiological events، plays crucial role in many pathological conditions such as tumor growth and metastasis. Recent studies indicate anticancer and antitumor properties of saffron against human cancers. Many processes are affected by Electromagnetic Field (EMF) and its effect on proliferation and gene expression were examined. In this experimental study، the synergic effects of saffron and EMF on VEGFR2 gene expression in MCF7 cells were investigated. Saffron was extracted using freeze dryer. MCF7 cells were grown in RPMI1640 medium supplemented with 10% FBS and incubated at 37C with 5% CO2. After 24 hr cells were treated with saffron extract at concentrations of 100، 200، 400 and 800 μg/ml. Forty eight hr after treatment all flasks were exposed with EMF (50 Hz، 0. 004 T). Then total RNA was extracted and cDNA was synthetized using specific primer. Synthetized products were analyzed by Real Time PCR to determine expression level of VEGFR2. Data were analyzed by SPSS (ANOVA & Tukey). Critical inhibitory effect on VEGFR2 gene expression was 20% at 400 μg/ml. Synergic use of EMF and saffron extract showed most reduction (38%) at 100 μg/ml. On the other hand synergic use of 200، 400 and 800 μg/ml saffron aqua extract and EMF decline noticeably the VEGFR2 level of gene expression to 29، 35 and 36%، respectively. EMF itself also reduced VEGFR2 up to 25% in comparison with control group which is remarkable at p<0. 001. Results indicate a decrease in the expression of vascular endothelial growth factor receptor in the treated samples with saffron extract compared to control. This reduction in VEGFR2 level induced by synergic treatment of saffron and EMF which reveals induction of inhibitory effects of saffron on angiogenesis and could be also considered as a promising chemotherapeutic agent in breast cancer treatment.