mehran miroliaei

In an effort to find effective medicines for the treatment of diabetes, the efficient mechanisms and lack of side effects of herbal medicines have made them the main candidates for regulating blood sugar levels and reducing the side effects of the disease. This search will be based on the discovery of digestive enzymes (α-amylase and α-glucosidase) inhibitors from natural sources and the ways to reduce high blood sugar levels. These enzymes are at the forefront of increasing blood glucose levels because they facilitate the digestion of food polysaccharides into smaller monosaccharide in small intestinal wall. Currently, the arsenal of inhibitors approved for this purpose is limited to veglibose, miglitol and acarbose. Despite the ability to reduce glucose absorption their widespread clinical use is limited due to the occurrence of gastrointestinal ailments. Given the efficacy of various natural compounds in alleviating diabetes symptoms, this review aims to assess the inhibitory potential and mode of action of some phytochemicals on intestinal digestive enzymes. Such an exploration seeks to unveil novel and healthful anti-diabetic agents based on the inhibition of digestive enzymes.

Given the prevalence of diabetes and the increasing number of diabetics, it is essential to find medicines to decrease the chronic complications of diabetes. Several studies have demonstrated that chronic hyperglycemia and its complications are directly related to protein glycation. Thus, identifying natural inhibitors to stop glycation of proteins may play a crucial role in managing the chronic complications of diabetes. Currently, various natural and synthetic compounds with anti-glycation attributes have been reported. The use of natural compounds in herbs (medicinal and non-medicinal) may be of particular importance due to fewer side effects and a wide range of therapeutic properties. Accordingly, this mini-review provides a list of common natural medicines and synthetic compounds with anti-glycation activity. As well, it provides brief information on the formation of advanced glycosylated end products (AGEs), their side effects, and glycation prevention mechanisms.

Diabetes mellitus is a metabolic syndrome characterized by elevated blood glucose. The α-glucosidase enzymes that are found in the small intestine are responsible for the hydrolysis of carbohydrates. The aim of this study was to Bioinformatics evaluation of T.foenum active compounds in suppression of α-glucosidase enzyme.

This study was a descriptive-analytical method. For this purpose, the compounds separation of Trigonella foenum were first downloaded from PubChem database and then the α-glucosidase enzyme file was obtained from PDB database. The toxicity class of compounds and the Lipinski rules were predicted by Toxtree & Protox II and the Swiss ADME server, respectively. Finally, molecular docking and enzyme interaction with the compounds in Trigonella foenumwere performed by AutoDock Tools 1.5.6 and Molegro Virtual Docker 6.0. Interaction results were also analyzed using Discovery Studio 3.5 & Ligplot 2.1 software.

The results indicated that all selected of compounds in Trigonella foenumwere in follow with Lipinskichr('39')s rules, proper binding energy, and lack of toxicity were appropriate options for α-glucosidase inhibition. But among these compounds, Vitexin had the lowest binding energy and the most inhibitory effect on the α-glucosidase enzyme, with -4.8 kcal/mol. These compounds also had lower binding energy than standard inhibitor (Voglibose).

From the results of this study, it can be concluded that among the most important compounds in Trigonella foenum, the Vitexin compound power inhibitor that due to more hydrogen and hydrophobic interactions with the α-glucosidase enzyme active site.

Aldose reductase plays an important role in chronic diabetes and can cause tissue complications. Controlling the activity of aldose reductase provides an important strategy in controlling the complications of diabetes. Flavonoids are important inhibitors of aldose reductase. This study aimed to investigate the inhibitory nature of flavonol compounds in fruits and plants on aldose reductase activity by bioinformatic methods.

This study was descriptive-analytical. First, the structure of flavonol compounds, Ranirestat, and enzymes were downloaded from ChemSpider and PDB databases, respectively. Ranirestat was used as the standard drug. The pharmacokinetic properties and toxicity of the compounds were predicted by Swiss ADME, Protox servers, and Toxtree2.5.4 software. Then, to interact the compounds with the enzyme structure, molecular docking was used to method AutoDock Tools 1.5.6. Finally, the results were analyzed using Discovery Studio 3.5 software.

The results showed that all flavonol compounds were desirable in pharmacokinetic properties and lacked toxicity. All compounds were able to inhibit aldose reductase enzymes with different PDB codes. However, among these compounds, the combination of Rutin with average docking energy of 191.21 kcal/ mole (P-value <0.05) had stronger binding energy than the standard Ranirestat drug.

From the results of this study, it can be concluded that among the selected flavonol compounds, the Rutin compound showed stronger inhibitory activity than other flavonol compounds due to its interaction with important amino acids in the active site of the enzyme. As a result, further study of this compound in vivo and in vitro environments can be used as a potential candidate to inhibit the enzyme aldose reductase and ultimately prevent chronic complications of diabetes.

Diabetes is one of the most common metabolic disorders. Alpha-amylase plays an important role in the development of diabetes by breaking down polysaccharide. Therefore, the search for natural inhibitor for α-amylase is of particular importance. Therefore, the aim of this study was to investigate the inhibitory effect of flavanone compounds on α-amylase enzyme by bioinformatics method.

This study was performed in the computer environment (Bioinformatics). For this purpose, the structure of flavanone compounds and α-amylase was downloaded from PubChem & Protein Data Ban database, respectively. Then, the drug-like parameter and physicochemical properties of flavanone compounds were investigated by Zink database and the Swiss ADME server, respectively. Then, in order to interact the compounds with α-amylase, one molecular docking software AutoDock Tools 6.0 was used. Finally, the results were analyzed using Discovery Studio 3.5.

The results showed that among the selected flavanone, naringenin compound was more desirable in terms of drug-like and physicochemical properties. Also, the result of molecular docking showed that the naringenin compound with a binding energy of -4.9 kcal/mol had the highest inhibitory effect on the α-amylase.

From this study, it can be calculated that naringenin compound shows more inhibitory ability due to its proper placement in the active site of α-amylase enzyme and interaction of key amino acids. By further investigation of this natural compound in In vivo & In vitro, it can be used as a natural inhibitor for the inhibition of α-amylase and the prevention of diabetes.

Salinity is a major environmental stress that causes severe damage to crops and horticultural crops and also reduces plant biodiversity. In order to study the resistance to salinity during seed germination and early growth, 18 strains of wheat cultivars (Triticum aestivum L.; Including car, non-indigenous and hybrid carcasses) were investigated at four levels of control, 100, 200 and 250 mM sodium chloride. The experiments were conducted with a complete randomized block design in three replications in the research laboratory of department of Biology, University of Isfahan, 2015. The results showed that with increasing NaCl level in the culture media, germination indices were affected with more intense. Comparison of mean traits showed that different cultivars of wheat had different reactions. These cultivars were classified in two groups, so that Sorghtoghm, Hamon, Sivand, Bezostaya, Sepahan and Roshan were placed in resistant groups and Ghods, Guspard, Karaj and Navid cultivars were placed in sensitive and low dendrogram groups. Sorghtoghm and Ghods were shown to have the highest and lowest index for germination stress index (GSI) and salinity tolerance index (STI) respectively. The results showed that the different levels of salinity had significant effects on germination percent, germination rate, seedlings stem and root length, root, and shoot dry weight. All measured variables showed a significant decrease with increasing salinity level.

From biomedicine point of view, the application of medicinal plants to prevent and treat diabetes via intervention in biomolecule degradation mechanisms, is an important issue. Achillea L. belongs to Asteraceae family and have traditional properties with different applications in folk medicine. A. pachycephala is one of the valuable species of Achillea which has been considered as a rich source of antioxidants. The present study aimed to investigate the effect of various phenological stages including five leaves appearance, sprouting, 50% and 100% flowering on antiglycative and antioxidant activity of A. pachycephala flowers extract. Antioxidant activity were evaluated based on 1,1-diphenyl 2-picrylhydrazyl (DPPH) and reducing power (FTC) methods. The inhibitory effect of the extracts was determined in albumin glycation model using Congo red binding and brown staining assays. The results showed that the extracts collected at 50% flowering stage had the highest antiglycative potential, the highest amounts of total phenols, total flavonoids and antioxidant activities. Besides, a direct correlation between antioxidant and anti-glycative activity was suggested. Most likely, A. pachycephala prevents albumin glycation originating from its strong antioxidant properties, therefore it can be considered as a good candidate for reducing complications of diabetes.

Reducing the complications associated with the healing of wounds and speeding up the wound healing process are among the topics of research for surgery science. There is a public interest in identifying efficient and cost-effective medicines, particularly herbal products. This study was aimed to investigate the effect of hydro-alcoholic extract of Mamisa plant (Khashkhash Bahri) on wound healing and to compare its effect with that of tetracycline in adult male rats.
Methods and materials: In this study, 30 male albino rats were randomly divided into three groups of 10 rats. After wounding the rats in a similar condition, the effects of Mamisa plant and tetracycline ointment on healing noninfectious wounds were examined. Accordingly, a control group was also considered. After seven and fourteen days of treatment using the plant extract and tetracycline ointment, we examined the trend of wound healing process in the dermis and epidermis layers in all groups. Compared with the control group, the herbal ointment group showed a significant difference (p Considering the results, it seem that the hydro alcoholic extract of Mamisa plant, because of its alkaloid and flavonoids compounds, speed up the wound healing in the dermis and epidermis layers.

A progressive computational analysis of available sequence and crystal structure data was used to identify functionally and structurally important residues in medium-chain Alcohol dehydrogenases super family throughout evolution. Altman and Gretsine core finding method was used to identify a core set of atoms with low structural variability. With further analysis of core regions in ADHs (solvent exposure and number of contacts) and also sequential analysis, we could infer the common properties of highly conserved positions in ADHs. The sequential analysis was done with respect to some special properties of amino acids in order to derive the level of conservation. The core structure analysis was re-judged in light of sequential analysis. Sixty percent of the core positions correspond to the highly conserved positions that were found by the sequence analysis.It seems that the core positions in ADHs are responsible for the maintenance of structural integrity and also contribute to the active site. Location of rigid parts of structures in catalytic domain may help to minimize the thermal fluctuation effect on substrate binding and would probably keep the structure in a good condition. Our data supports that it is possible to define patterns of conservity in some important region in sequence of Alcohol dehydrogenase and explain patterns of structural stability which are necessary for overcoming throughout evolution.

Nonspecific lipid transfer protein 2 (nsLTP2) is among prolamin super family, capable of transferring various lipophilic molecules such as sterols. Despite advantages of the protein in drug delivery systems, there are some reports on its allergenic properties. Comparative sequence analysis displayed at least 51% homology in different species of rice-nsLTP2. The aim of the current study was to evaluate gene sequence and protein structure analysis of Iranian rice-nsLTP2, by means of bioinformatics tools, to improve its potential capability for drug delivery systems. Gene sequence homology assessment revealed resemblance mostly with Japonica species (Accession NO. NP-0011048723.1). The presence of various specific sites and motifs in the structure of nsLTP2 were determined by sequence analysis. Preliminary studies demonstrated four antigenic determinants in the primary sequence of the protein, which three of them retain after post translational modification. Moreover, sequence assessment revealed that those residues contributing in the protein cavity are conserved totally among Japonica and Iranian species. Modeling approaches were used to determine the influence of deletion of allergenic sites on protein affinity toward hydrophobic ligands. The obtained results revealed the efficacy of nsLTP2, as a drug carrier protein, could be improve by appropriate swap at specific residues and elision of allergenic sites.

NSLTPs are plant proteins that are subdivided into nsLTP1 (9kDa) and nsLTP2 (7kDa) according to the molecular weight. These highly stable proteins can protect drugs against oxidation or degradations. Rice NsLTP2 due to the inherent ability to cross biological membranes and bind to steroid compounds (including some pharmaceutical compounds) have potential application for use in drug delivery systems. After designing of mutagenic primers (contains a mutation of (Y45W)) the method of SOE-PCR (splicing by overlap extension-polymerase chain reaction) is used to site directed mutagenesis in the gene encoding the rice nsLTP2. Wild type nsLTP2 was cloned in to the PGEX6p2 vector. This wild type plasmid was used as a template for constructing the mutant fragment and designed mutant primers are used for amplification of mutant fragments. Mutant rice nsLTP2 are expressed in the pET32a vector in to the E. coli BL21 (DE3) strain. After the amplification of the mutant fragments and confirmation of mutations generated by sequencing، the mutant constructs cloned into the pET32a vector and the fusion mutant protein with a His-tag was purified by Ni-NTA Affinity chromatography column. The purpose of this study is Site directed mutagenesis to production the mutant Iranian rice-NSLTP2 (Oriza sativa) protein that contains Y45Wmutation. We hope that، in the future، these findings could contribute that this protein could be used to create biosensors or act as drug carriers.

The ability to reducing death due to apoptosis and maintaining extensive levels on cell viability under serum free media in cell culture are important subjects in production of recombinant proteins. Insulin-like growth factor-I (IGF-I), is the growth factor of choice for mammalian cell proliferation in serum-free culture. In addition to its mitogenic activity, it has antiapoptotic activity protecting cultures from diverse death inducing stimuli. In this study, the effect of different concentrations of IGF-I examined on CHO–K1 (Chinese hamster ovary K1 cells) cell line for 24 and 48 hours.

In this experimental study, the cell line was cultivated in DMEM supplemented with 10% fetal bovine serum (FBS). Apoptosis process was induced in cells by methotrexate, serum was removed and then 10-50 ng/ml concentrations of IGF-I were added. Apoptosis was assessed by caspase 3 detection kit and cell proliferation and viability determined by MTT assay.

Caspase 3 activity decreased significantly by increasing concentrations of IGF-I. In the highest concentration of IGF-I (50 ng/ml), 1.7 and 1.4 equal decreases of caspase 3 activities, compared with negative control group, were noted after 24 and 48 hours, respectively. It confirmed antiapoptotic activity of growth factor to maintain viability and protect cultures from apoptotic inducing stimuli (methotrexate).

IGF-I, as antiapoptotic factor, decreased programmed death of CHO-K1 cells in apoptotic induced conditions.

In the present study, we optimized the experimental conditions using pET-15b expression vector to obtain large amounts of Taq DNA polymerase.

Correct framing of the gene in the expression vector pET-15b and its orientation were analyzed by digestion and sequencing. Production of Taq DNA polymerase in Escherichia coli BL21 (DE3) cells was induced by incubation with different concentrations of IPTG. Optimum production occurred with the addition of 1 mM IPTG for 2 h. The activity of the obtained enzyme was measured by comparing the intensities of the produced DNA bands in PCR reactions.

Recombinant plasmid containing taq polymerase gene was confirmed by restriction digestion and DNA sequencing. Purified protein was identified by Western blotting. Optimum condition for the production of the enzyme was induction with 1 mM IPTG for 2-3 h. Addition of NP-40 increased enzyme stability.

We expressed the recombinant Taq DNA polymerase in E.coli using a T7-based promoter system and obtained an active and stable enzyme.

The p75 neurotrophin receptor (p75NTR) is a 75-kDa glycoprotein and the first identified member of the TNF receptor superfamily. p75NTR is initially expressed very early in embryogenesis in cells derived from all three germinal layers but becomes progressively restricted to specific cell populations such as CNS and PNS. The p75NTR expression is induced after pathologic conditions in a wide variety of cell types. p75NTR is a versatile protein, but its function in different cell types has remained enigmatic. One of the reasons for the lack of clarity regarding the function of p75NTR is that it does not signal by traditional pathways; it has no kinase activity nor is it linked to a G-protein-coupled pathway. p75NTR has no intrinsic enzymatic activity so it depends on its interaction with several adaptor proteins to signal the cascade for neuronal survival or death. The various functions of p75 depend on the type of ligand bound to it (neurotrophins or pro- neurotrophins), the cell type in which it is expressed and the presence or absence of Trk receptor and Interaction with other receptors. This mini-review explores the function, gene expression, interactions with co-receptors and signaling pathways of p75NTR in the nervous system.

Molecular chaperones are characterized by a general behavior, arresting the exposed hydrophobic surfaces of denaturing substrate proteins. In the present study, the capacity of β-caseins (β-CN) from camel and bovine milk in suppression of thermal aggregation process of apo-yeast alcohol dehydrogenase (YADH) was assessed. Apo-I enzyme was prepared by removal of the structural zinc; while apo-II-protein was obtained by depleting conformational and catalytic zinc atoms. Fluorescence spectroscopy using ANS probe revealed greater hydrophobic surface in apo-II ADH. Considerable decrease in aggregation of the heat treated protein molecules was observed upon exposing to β-CNs (camel, bovine). Bovine β-CN afforded more adverse effects on thermal aggregation. A direct correlation between casein’s chaperone activity and structural stability of the substrate proteins was displayed. Moreover, an association between casein source and chaperone-like activity is suggested.

Plant nonspecific lipid transfer proteins (nsLTPs) are divided into nsLTP1 and nsLTP2. The existence of an internal hydrophobic cavity, is a typical characteristic of nsLTPs that serves as the binding site for lipid substrates. In this communication a simple, rapid and low-cost alternative method was developed for purification of nsLTP2 from rice paddy. After extracting, final supernatant was loaded on CM-Sepharose column, which had previously equilibrated with 0.05 M Tris-HCl buffer, pH 8. Bounded proteins were separated using a linear gradient of 0-0.5 M NaCl. Solution of separated proteins was dialyzed and applied on a Phenyl-Sepharose column which previously equilibrated with Tris-HCl 0.05 M, ammonium sulfate 1.5 M, EDTA 0.005 M and NaHSO3 0.3%, pH 8.4. Tris-Tricin SDS-PAGE of separated proteins, obtained from ionexchange column, showed multiple bands in the range of 2-20 kDa. Further purification using hydrophobic column resulted in single band of nsLTP2 at about 7 kDa, reflecting a purified sample in the gel.

Epitope tagging is a method of expressing proteins whereby an epitope for a specific monoclonal antibody is fused to a target protein using recombinant DNA techniques. The aim of this study was to sub-clone the peroxisomal protein (PEP) cDNA into a mammalian expression vector leading to the formation of a chimeric PEP-cDNA containing the FLAG epitope. The FLAG-PEP recombinant cDNA was constructed using the method of splicing by overlap extension polymerase chain reaction (SOE PCR) and inserted into the pUcD2SRaMCSHyg eukaryotic expression vector. To investigate the intracellular localization of the PEP protein that was linked to the FLAG tandem, the constructed plasmid was used for transient transfection of the Chinese hamster Ovary (CHO) cells. The CHO cells that were transfected with the recombinant plasmid showed peroxisomal localization of FLAG-PEP as was previously shown for catalase.