Iranian Journal of Biotechnology
Volume:15 Issue: 4, Autumn 2017

Use of Adipose Stem Cells (ADSCs), obtained easily in a relatively less invasive manner (abdominoplasty) and characterized by flow cytometry, is a classical approach in stem cell research and clinical aspects. Other techniques such as isolation of the cells from bone marrow aspirates (1) are rather more invasive. Further, it is pertinent to point out that growth rate, differentiation potential and functions are better in abdominal Adipose Stem Cells (ADSCs), in comparison with those isolated from the Visceral Omental Pads. Such changes are attributable to these cells retaining the memory of their sites of origin (2). Furthermore, their other features of having the potential to differentiate into a number of lineages coupled with their immunomodulatory, angiogenic, anti-inflammatory and anti-apoptotic effects puts ADSCs as the prominent cell type in stem cell research and clinical aspects. The authors have chosen alginate (specific M/G ratio)-gelatin composite hydrogel, with favourable charge properties, to demonstrate an increased growth rate and differentiation potential (50:50 ratio better than the 70:30 ratio) (1) Gelatin is widely accepted as a biocompatible, biodegradable, cost-effective substratum for cell growth...

Different concentrations of the simple carbon substrates i.e. glucose, fructose, and sucrose were tested to enhance the performance of the mediator-less double chamber microbial fuel cell (MFC).
The power generation potential of the different electron donors was studied using a mesophilic Fe (III) reducer and non-fermentative bacteria Pseudomonas aeruginosa-isolated from municipal wastewater.
A double chamber MFC was operated with three different electron donors including glucose, sucrose, and fructose. Substrate utilization pattern was determined through chemical oxygen demand (COD) removal rate and voltage generation. In addition, electrochemical, physicochemical, and microscopic analysis of the anodic biofilm was conducted.
P. aeruginosa was proven to effectively utilize hexose and pentose sugars through anode respiration. Higher power density was generated from glucose (136 ± 87 mWm-²) lead by fructose (3.6 ± 1.6 mWm-²) and sucrose (8.606 ± mWm-²). Furthermore, a direct relation was demonstrated between current generation rate and COD removal efficiency. COD removal rates were, 88.5% ± 4.3%, 67.5% ± 2.6%, and 54.2% ± 1.9% with the three respective sugars in MFC. Scanning electron microscopy (SEM) demonstrated that the bacterial attachment was considerably abundant in glucose fed MFC than in the fructose and sucrose operated MFC.
This study has revealed that electron donor type in the anodic compartment controls the growth of anodic biofilm or anode-respiring bacteria (ARB).

Human alpha 1-antitrypsin (AAT) is a monomeric glycosylated protein; it is the potent inhibitor of a whole range of serine proteases and protects tissues against their destructive effects. The human plasma-derived AAT, which is currently used to augment the AAT level in patients, is limited due to high cost and source limitation. Recombinant production of AAT can be considered as a potential alternative.
This study aims to develop and optimize a new chemically defined medium based on an elemental analysis of the yeast Pichia pastoris for an efficient culture of the recombinant yeast-producing secretory AAT.
Material and An elemental analysis of Carbon (C), Hydrogen (H), Nitrogen (N), Sulfur (S); CHNS in its abbreviated form, and metallic elements was performed to determine the exact molecular constituent of the P. pastoris. The medium components were selected according to the obtained formula; they were optimized by the response surface methodology (RSM). The grown yeast cell was measured at the end of 18 h glycerol batch culture. The amounts of AAT production and elastase inhibitory capacity (EIC) were measured at the end of three days’ methanol feeding.
The optimized medium compositions consist of glycerol (40 g.L-1), KH2PO4 (24.78 g.L-1), NaCl, (0.88 g.L-1), MgSO4.7H2O (1.95 g.L-1), (NH4)2SO4 (22.76 g.L-1), and trace elements (20 mL.L-1). The presented quadratic models show that KH2PO4 and (NH4)2SO4, are the most abundant ones in the P. pastoris biomass and have the greatest effect on the cell growth, EIC, and AAT protein production responses.
According to the results of this study, it can be concluded that the characterizing cell composition formula could be considered as an appropriate method to design culture media in order to improve cell growth and productivity. Compared to the common P. pastoris chemically defined media, FM22 and BSM, production of AAT protein increased by 1.5 and 1.4 times, respectively, in this new medium.

The need for more cost-effective compounds is imperative because the demand for prebiotic compounds is ever on the rise.
The focus of this study is the purification of the endoxylanase from Bacillus pumilus B20 and its application in a cost-effective production of the prebiotic xylooligosaccharide (XOS) syrup having a high concentration of oligosaccharides.
Theextracellular endoxylanase was purified using ammonium sulphate fractionation, DEAE anion exchange, and Sephacryl gel filtration chromatography. The enzymatically produced XOS was used in the preparation of XOS syrup adopting the method of ultrafiltration with 10 and 3 kDa molecular weight cut-off (MWCO) membranes. Culture-dependent technique for the bacterial enumeration using selective probiotic microorganisms in an in vitro analysis was employed to confirm the prebiotic nature of XOS syrup.
The molecular mass of the purified xylanase (XylB) was found to be approximately 85 kDa with the optimum pH and temperature of 6.5 and 60 °C, respectively. XylB hydrolyzed the xylan and produced short-chain xylooligosaccharides (XOS). At the end of the two-step ultrafiltration process, the hydrolysate was refined to form XOS syrup (44.4%) consisting of XOS with a degree of polymerization (DP) between 2 and 5, and >5. Among all the tested probiotic strains, Lactobacillus brevis exhibited maximum growth in the presence of 0.5% XOS syrup with a specific growth rate of 1.2 h-1.
Through this study, we have identified a method to produce XOS syrup that can be used as an effective prebiotic supplement for the growth of several probiotic strains. Human gut probiotics was used as a model system for in vitro analysis of prebiotic oligosaccharide XOS, but for further confirmation of the prebiotic activity, in vivo feeding studies using animal models are needed to be carried out.

Mesenchymal stem cells (MSCs) are multipotent cells able to differentiating into a variety of mesenchymal tissues including osteoblasts, adipocytes and several other tissues.
Differentiation of MSCs into fibroblast cells in vitro is an attractive strategy to achieve fibroblast cell and use them for purposes such as regeneration medicine. The goal of this study was investigate the simulated microgravity effect on differentiation of Adipose Derived Stem Cells (ADSCs) to fibroblasts.
To fibroblast differentiation 100 ng.mL-1 of connective tissue growth factor (CTGF), and for simulation microgravity, 2D clinostat was used. After isolation the human ADSCs from adipose, cells were passaged, and at passages 3 they were used for characterization and subsequent steps. After 7 days of CTGF and simulated microgravity treatment, proliferation, and differentiation were analyzed collectively by MTT assay, quantitative PCR analyses, and Immunocytochemistry staining.
MTT assay revealed that CTGF stimulate the proliferation but simulated microgravity didn’t have statistically significant effect on cell proliferation. In RNA level the expression of these genes are investigated: collagen type I (COLI), elastin (ELA), collagen type III (ColIII), Matrix Metalloproteinases I(MMP1), Fibronectin 1 (FN1), CD44, Fibroblast Specific protein (FSP-1), Integrin Subunit Beta 1 (ITGB1), Vimentin (VIM) and Fibrillin (FBN). We found that expression of ELN, FN1, FSP1, COL1A1, ITGB1, MMP1 and COL3A1 in both condition, and VIM and FBN1 just in differentiation medium in normal gravity increased. In protein level the expression of COL III and ELN in simulated microgravity increased.
These findings collectively demonstrate that the simulated microgravity condition alters the marker fibroblast gene expression in fibroblast differentiation process.

Obesity, a global health problem and a chronic disease, is associated with increased risk of developing type 2 diabetes and coronary heart diseases. A wide variety of natural remedies have been explored for their obesity treatment potential.
The anti-adipogenic effect of ginsenoside Rg5:Rk1 (Rg5:Rk1) on 3T3-L1 mature adipocytes was investigated.
To elucidate the anti-obesity effect of Rg5:Rk1, a mixture of protopanaxadiol type ginsenosides isolated from Panax ginseng Meyer in a 3T3-L1 adipocytes. In order to determine the anti-obesity effect of Rg5:Rk1, based on oil Red O Staining, triglyceride (TG) content in adipose cells was assessed. Furthermore, to elucidate the possible mechanism of Rg5:RK1 effect on lipid accumulation, mRNA and protein expression analyses of adipocyte markers such as STAT3, PPARγ, CBEPα and ap2 were carried out.
Rg5:Rk1 treatment showed an inhibition of lipid droplet accumulation and decrease of TG content. In addition, expression of STAT3, PPARγ, CEBPα and ap2 were decreased in a dose dependent manner. Similarly, the Rg5:Rk1 treatment reduced PPARγ and CEBPα protein expression.
Rg5:Rk1 treatment exhibits anti-adipogenic activity by down-regulation of the STAT3/ PPARg/CEBPa signaling pathway in 3T3-L1 adipocyte cell line.

Potyvirus-based virus-induced gene silencing (VIGS) is used for knocking down the expression of a target gene in numerous plant species. Sugarcane mosaic virus (SCMV) is a monopartite, positive single strand RNA virus.
pBINTRA6 vector was modified by inserting a gene segment of SCMV in place of Tobacco rattle virus (TRV) genome part 1 (TRV1 or RNA1) and the two nonstructural proteins of TRV2(RNA2).
SCMV construct was inoculated into 3-4 weeks Nicotiana benthamiana plant leaves either by using a needleless syringe or applying pricking with a toothpick.
The construct (SCMV-RNA2) successfully induced post-transcriptional gene silencing (PTGS) of the target genes GFP and ChlI through agroinoculation proving that SCMV is a substitute of the RNA1, which plays a pivotal role in the systemic gene silencing. 2-3-weeks of post inoculation, target genes’ silencing was observed in the newly developed non-inoculated leaves.
The newly developed construct expresses the knocked down of the endogenous as well as exogenous genes and only four weeks are required for the transient expression of the gene silencing based on SCMV-VIGS system.

Recent theranostic (therapeutic or diagnostic) applications of tellurium nanoparticles have attracted a great interest for development of different methods for synthesis of this valuable nanostructure, especially via biological resources.
In the present study, the antimicrobial and antioxidant effects of the tellurium nanorods (Te NRs) biosynthesized by a bacterial strain Pseudomonas pseudoalcaligenes strain Te were evaluated.
The antimicrobial effect of Te NRs and potassium tellurite against different bacterial and fungal pathogens was assessed by microdilution method. Furthermore, the disk diffusion method was used to evaluate the antibacterial effect of the biogenic Te NRs and potassium tellurite against methicillin-resistant Staphylococcus aureus, alone or in combination with various antibiotics. Also, the biogenic Te NRs were investigated for antioxidant activity using 2, 2-diphenyl- 1-picrylhydrazyl (DPPH) scavenging activity and reducing power assay.
Transmission electron micrograph (TEM) of the purified Te NRs showed individual and rod-shaped nanostructure (~22 nm diameter by 185 nm in length). Based on the data obtained from both microdilution and disk diffusion method the K2TeO3 exhibited a higher antibacterial and antifungal activity compared to the Te NRs. The measured IC50 for the biogenic Te NRs (i.e. DPPH radical scavenging activity) was found to be 24.9 μg.mL-1, while, K2TeO3 has represented only 17.6 ± 0.8 % DPPH radical scavenging effect at the concentration of 160 μg.mL-1. The reducing power assay revealed a higher electron-donating activity for Te NRs compared to K2TeO3.
Based on the data obtained from both microdilution and disk diffusion method the K2TeO3 exhibited a higher antimicrobial and antifungal activity than Te NRs. Te NRs didn’t show the antibacterial effect against the tested bacterial strain: MRSA and showed an inhibitory effect and antibacterial activity of the effective antibiotics. However, more studies should be performed to explore the action mechanism of the produced biogenic Te NRs.

Green tea polyphenols (GTP) are known to have several health benefits. In spite of these benefits, its application as a therapeutic agent is limited due to some of its limitations such as stability, bioavailability, and biotransformation. To overcome these limitations, liposomal nanoparticles have been used as a carrier of the GTP.
Encapsulation of GTP to the liposomal nanoparticles in order to achieve a sustained release of the GTP and to determine the drug release kinetics and the mechanism of the release.
GTP encapsulated liposomal nanoparticles were prepared using phosphatidyl choline and cholesterol. The synthesized particles were characterized for their particle size and morphology. In vitro release studies were carried out, followed by drug release kinetics, and determining the mechanism of release. In vitro, antioxidant assay was determined following 2,2-diphenyl-1-picrylhydrazyl (DPPH) method.
Atomic force microscope (AFM) and high resolution scanning electron microscope (HR SEM) images showed spherical particles of the size of 64.5 and 252 nm. An encapsulation efficiency as high as 77.7% was observed with GTP concentration of 5 mg.mL-1. In vitro release studies showed that the loading concentrations of GTP were independent to the cumulative percentage of the drug release. GTP release by varying the pH and temperature showed a direct correlation between the release parameter and the percentage of drug release. The higher the pH and temperature, the higher was the percentage of the drug release. The release data showed a good correlation with Zero order kinetics and the mechanism of the release being anomalous mode. Radical scavenging activity of the released GTP showed a potent scavenging activity.
GTP encapsulated liposomal nanoparticles could be used as a delivery vehicle for achieving a sustained release.

The increasing use of nanoparticles (NPs) may have negative impacts on both organisms andthe environment.
The differential expression of mitochondrial manganese superoxide dismutase (MnSOD) gene in wheat in response to silver nitrate nanoparticles (AgNPs) and AgNO3 was investigated.
A quantitative Real-Time RT-PCR experiment was carried out with MnSOD gene using RNAs isolated from wheat shoots treated for 0, 2, 6, 12, and 24 h with 100 mg.L-1 of either AgNO3 or AgNPs.
The results of this study showed that both treatments cause changes in the expression pattern of the MnSOD gene. While 2 and 6 h following the beginning of the stress, MnSOD expression was up-regulated significantly, in response to AgNO3 (1.4 and 2.8 fold, respectively), in response to AgNPs, it was up-regulated significant only after 6 h (1.6 fold), compared with the control. The gene expression, after 12 h in response to AgNO3 and AgNPs were down-regulated significantly (0.7 and 0.8 fold, respectively), and in the next 12 h , the expression appeared to be similar to the control.
Exposure to both AgNPs and Ag ions led to a significant increase in MnSOD expression, but AgNO3 changed the MnSOD expression faster than AgNPs. Therefore, it is suggested that AgNO3 has greater penetrability and effectiveness.