محمدرضا خرمی زاده

The present study was performed to determine the effects of resveratrol as a natural polyphenol on growth and nutrition indices of zebrafish (Danio rerio). Resveratrol was performed over a period of 60 days on 120 adult zebrafish with approximate weight of 2 ± 0.3 g and length of 3 ± 0.5 cm, which were randomly divided into 2 main groups, each of which consisted of 4 subgroups (a total of 8 Treatment) were divided as follows: The two main groups under the influence of added glucose in aquarium water (+ G) and without glucose (-G) and each of these two groups into 4 subgroups (control = CTRL) , (resveratrol = RSV ) With doses of 20, 10 and 30 micromole per liter. Fish were randomly distributed in 5 pieces into 24 tanks (3 treatment in 3 replicant) with a water volume of 3 liters. At the end of the period, weight gain, feed conversion ratio, protein efficiency ratio, specific growth rate and body growth index and enzymes of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkannin phosphatase (ALP) were evaluated. According to the results, the highest feed conversion ratio belonged to the resveratrol 20 group under the influence of glucose (G + RSV30). The highest rate of protein efficiency ratio, specific growth rate and body weight gain belonged to the glucose-free control group and no significant difference was observed between treatments. Also, the treatment of resveratrol 20 group without glucose (G-RSV20) had the lowest amount of enzyme (ALP) and no significant difference was observed with other groups and the treatment of control group without glucose (G-CTRL) had the lowest amount of enzymes (AST) and (ALT) and a significant difference was observed with other groups (p <0.05). The results of this study indicate that resveratrol did not have a positive effect on growth indices and AST and ALT enzymes but improved ALP enzyme function.

Hyper-glycosylation is an approach to introduce new N-glycosylation consensus sequence(s) (Asn-Xxx-Ser/Thr three-peptide) into a protein primary amino acid sequences by site-directed mutagenesis which is followed by the attachment of a new glycan to the Asn residue located within the three-peptide sequence. Hyper-glycosylation has attracted lots of interest especially in the protein therapeutics industry. The attached glycan may improve the pharmacokinetic properties of the hyper-glycosylated priteins and increase their half-life in the bloodstream. In the current study, a new N-glycosylation site was introduced into N-terminal Gla domain of hFIX. Arg37 position of mature hFIX was targeted to be converted into Asn residue by site-directed mutagenesis using overlap extension PCR. Recombinant expression plasmids for native and mutant hFIX were constructed. The expression of the recombinant wild-type and mutant hFIX was analyzed in mammalian HEK293 cells using gradient SDS-PAGE and western blotting analysis. The results indicated in higher molecular weight for R37N mutant in compared with the native protein. The glycan attachment to R37N mutant was further confirmed by PNGase digestion and western blotting.

A novel nanocomposite-modified electrode based on reduced graphene oxide (rGO) decorated with crown-ether and gold nanoparticles (GNPs) on the surface of a glassy carbon electrode (GCE) was fabricated to investigate 5-HT determination.

The morphology of nanocomposite was characterized by scaning electron microscopy (SEM). Diabetic zebrafish was obtained by overfeeding via glucose. 5-HT was successfully determined in the presence of dopamine (DA), ascorbic acid (AA), urea, glucose, and L-tryptophan (L-Trp) by using electrochemical methods.

The nanocomposite exhibited satisfactory electrochemical catalytic activity for 5-HT determination using square-wave voltammetry (SWV). The electrochemical behavior of 5-HT at the nanocomposite/GCE displayed reasonable oxidation current and potential. The limit of detection (LOD) of 5-HT obtained from the real samples, containing the control and diabetic group by using the proposed system and HPLC method, was calculated to be about 0.3 and 0.1 µg/L, respectively. The prposed system also demonstrated high selectivity, reasonable sensitivity, and good stability and reproducibility for 5-HT sensing. The nanocomposite was applied for the determination of the biomarker 5-HT in the diabetic and control groups of zebrafish and displayed excellent recoveries about 93 and low relative error about 3% while compared with standard method.

It seems that the 5-HT level can be used for earlier diagnosis of diabetes.

Multiple sclerosis (MS) is considered as a common inflammatory disease of the human central nervous system (CNS). 4 amino pyridine (4-AP), a potassium channel blocker, is used widely in MS treatment to reduce fatigue and cachexia often experienced by the patients. The objective of this study was to get a better understanding of the mechanism of action of this drug, using the cell culture model. More specifically, we attempted to determine the effects of the drug on 1. The profileration of, and its cytotoxicity on, the neurons, and 2. The activity of MMP-9 in the neurons.

Considering the available reports on the wide range of 4-AP effects, we designed this study to investigate its possible role in proliferation or cytotoxicity of the MS cellular model, i.e. astrocytoma U373-MG, using the MTT assay technique. We also analyzed the effect of 4-AP on cell functionality as assessed by matrix metalloproteinase-9 (MMP-9) zymography.

The results showed that while 4-AP at a concentration of 0.1 and 1 mM has no significant cytotoxic effects on the treated cells, it has remarkable MMP-9 inhibitory effects (p<0.01). The proliferation analyses confirmed the stability of 4-AP effects on cell functionality.

On the whole, the results of the present study show the desirable effect of 4-AP on MMP-9 activity at non-cytotoxic doses, promising its further therapeutic applications.

Evaluation of Matrix Metaloproteinase 2 (MMP2) Activity in Contact Nickel Dermatitis Using Zymoanalysis in Comparison to Normal Individuals. R. Falak MSc*M.R. Khoramizadeh MD**M. Pezeshki MD*** P. Mansoori MD**** * Master of Seience of Immunology Iran University of Medical Seiences ** Associate Professor of Biotechnology Faculty of Health Tehran University of Medical Sciences *** Associate Professor of Immunology Faculty of Health Tehran University of Medical Sciences **** Professor of Dermatology Imam Khomeini Hospital Tehran University of Medical Sciences Background and objective Matrix Metaloproteinases (MMPs) produced mainly by fibroblasts play a major role in wound healing processes. This study aimed to investigate the MMP2 activity in dermal fibroblasts found in chronic contact nickel dermatitis lesions. Methods In order to study the role of MMP2 in contact nickel dermatitis fibroblast from patients and healthy individuals were cultured based on ex-plantation of skin. MMP2 activity was measured in fibroblast culture media using zymoanalysis. Zymograms were then analyzed by quantitative densitometry. MTT assay was also used to evaluate and compare proliferation capacity of fibroblasts in both cell cultures. Results The mean MMP2 activity 6-8 days after ex-plantation was significantly higher in patients fibroblasts than in normal individuals(170±9.2 and 134.3±5.9 in patients and normals respectively.

Trichophyton verrucosum (T.verrucosum) is one of the dermatophyte fungi which invades the skin of human and animals particularly cattle. Several properties of this fungus have been investigated so far. However a few studies were carried out in the field of molecular biology of this fungus. In the present study we tried to identify the chaperone proteins related to the 70 kDa heat shock proteins (HSP70) in this fungus and study of its gene regulation under heat-stress conditions. 

Pairs of 21 and 20 nt primers were designed from highly conserved regions of the similar genes in other fungi. Mentioned primers were utilized in PCR by using isolated genomic DNA and cDNA of T.verrucosum whereas the PCR fragments were then sequenced. 

Eventually, 2217 nucleotides have been sequenced from this new gene which has two introns and encodes a polypeptide with 654 amino acids. Sequences comparison in gene data banks (NCBI, NIH) for both the complete DNA and its deduced amino acid revealed significant homology with members of the eukaryotic 70 kDa HSP family. Further investigation revealed that the expression of this new gene increases under heat-stress conditions. 

This new gene has similarity with other genes of the same family in other organisms. This protein could efficiently protect the microorganism against heat stress.