نشریه زیست فناوری دانشگاه تربیت مدرس
سال دوازدهم شماره 3 (پیاپی 30، تابستان 1400)

In recent years, two-dimensional nanomaterials are widely used in biomedical applications. Graphitic carbon nitride has been significant not only due to good biocompatibility but also because of its application in the bio-imaging, diagnosis, and treatment of cancer. The purpose of this study was to investigate the effect of graphitic carbon nitride on viability of Saos-2 cancer cell line.

Graphitic carbon nitride was synthesized by adding melamine to hydrochloride acid and its physicochemical properties and structure was evaluated using FTIR, XRD, and Raman spectroscopy. Synthesized graphitic carbon nitride (50, 100, 200, 400, and 800 μg/mL) was added on Saos-2 cells and fibroblasts in two periods of 48 and 72 hours. The survival rate of the cells was investigated by MTT.

The results of FT-IR and XRD tests showed the structure and bonds of the synthesized compound, its accuracy and purity, respectively. Raman spectroscopy results also showed the graphitic content of the synthesized product. After passing 48 hours of the exposure of the cells to synthesized graphitic carbon nitride, the level of cell survival in the group that received 800 µg/mL was decreased to nearly 80% compared to the control (untreated) group.

The synthesized compound in this study might be considered as an appropriate option in cancer related researches.

Nowadays, aromatic componds are widely used in food, pharmaceutical, cosmetic and chemical industries. Due to the growing tendency of consumers to use natural products, biotransformation by microorganisms is an interesting method for the production of aromatic compounds. Gamma-decalactone is a cyclic aroma with a peach-like flavor. The non-conventional yeast Yarrowia lipolytica can bioconversion the inexpensive castor oil substrate into the valuable gamma-decalactone. The process begins with the hydrolysis of castor oil by the lipase to ricinoleic acid, then continues by shortening the chain by beta-oxidation, and finally ends by lactonization. In this study, gamma-decalactone production was optimized via Response surface methodology (RSM) by a mutant strain of this yeast with the ability to produce high amounts of lipase. For this purpose, castor oil, yeast extract, peptone and pH were studied as factors at five levels. Based on statistical analysis of the relationships between the experimental variables, a mathematical model was obtained for the governing relationships between the experimental variables. Based on the results, the best values were obtained for castor oil 35 ml/l, yeast extract 6 g/l, peptone 8.5 g/l and pH 4. To validate the mathematical model, the proposed values were tested and 126 mg/L of gamma-decalactone was produced by the yeast strain, which shows a 46% increase compared to non-optimal conditions. The results of this study can be used to make cost-effective production of gamma-decalactone from castor oil by microbial biotransformation process.

Today, the use of medicinal plants in the cancer treatment due to less side effects has been considered. Silybum marianum is a medicinal herb of Asteraceae, which is used in the treatment of liver diseases and gallbladder diseases, cancer, cardiovascular diseases. The encapsulation of bioactive materials in nano-liposomes is an effective approach to regulate drug release, increase stability, protect them from environmental reactions, reduce volatility, and increase its effects. The aim of this study was encapsulation of the extract of Silybum marianum in to liposomes and to evaluate the physico-chemical in order to effect on liver cancer cells

In this study, extract of Silybum marianum was prepared by Soxhlet method. Liposomal vesicles were prepared by thin-film hydration method and the extract of Silybum marianum was loaded. Finally, the nanoparticles were assayed for encapsulation efficiency, release profile and physicochemical properties such as particle size, zeta potential, morphology, and FTIR.

Nanoliposome containing Silybum marianum extract had 63.37% encapsulation efficiency and size 122 nm zeta potential -13.1 and the dispersion index 0.197. The release of herbal extract of Silybum marianum was controlled. There is no chemical interaction between the extract and the liposome and is morphologically homogeneous and had a spherical structure.

The results of this study show that the extracts of Silybum marianum can be encapsulated in appropriate size and function in nanoliposomal forms, so liposomes are a suitable carrier for the Silybum marianum extract.

Different defense pathways in plants evolved in reaction to pathogens. The main aim of this study was to investigate the mechanism of action of glyphosate in resistance induction to bacterial phytopathogens. To do so, glyphosate at an optimal concentration of 1.8 mg / l was used on transgenic potato, to induce resistance to two strains of pathogenic bacteria (21A of Pectobacterium atrosepticum and ENA49 of Dickeya dadantii). It was been shown that plant defense responses to pathogens can be stimulated by treatment plants at an optimal concentration of glyphosate. Transgenic potato leaves infected with potato pathogenic bacteria, and then treated with glyphosate showed a high level of expression of pathogenesis-related genes (PR-2, PR-3, PR-5), especially PR-2 gene and defense response genes (HSR-203j, HIN1), especially HSR-203j gene. The expression of PR-2 gene in leaves infected with these two bacteria were 1.5 and 2.9 times, for PR-3 gene 1.7 and 1.7 times, for PR-5 gene to 1.3 and 1.5 times and expression of HSR-203J gene to 2.5 and 2.4 times and - HIN1 gene to 1.7 and 1.7 times, with Dickeya dadantii and Pectobacterium atrosepticum infection, respectively. The expression of these genes in control samples didn’t significantly change. The results showed that there was a significant difference between the expression of genes in the experimental and control samples (plants treated by glyphosate compared to untreated plants). The results showed that the treatment of plants by glyphosate can induce a systemic acquired resistance to phytopathogens by inducing proteins and defense response genes.

Rye is one of Iran's most important crops, known as Secale, belonging to the Poaceae. In this research, genetic diversity of 39 families of rye populations from different regions of Iran, the USA, and the Soviet :union: was evaluated with the ISSR marker. The results showed that 8 ISSR initiators produced 48 bands which included 18 polymorphic bands (37.5% polymorphism). The mean of polymorphic information content (PIC) and marker index value (MI) for ISSR primers was 0.15 and 7.2 respectively. The highest PIC (3.0) was related to primer 5+6 and the highest MI (0.96) reached for primer 1+6. After observing polymerase chain reaction products on an agarose gel and scoring DNA bands, the analysis was performed with NTSYS software. The cluster analysis dendrogram of UPGMA and Jaccard's similarity coefficient divided the rye populations into 9 groups, the results were compiled with grouping by principal component analysis. The results of analysis of Molecular Variance indicated an in-species variation more than inter-species variation. The mean Nei genetic variation (h) was 350 and the mean of Shannon index (I) in rye species was 523, which indicates a relatively good variety within species. The results showed that the ISSR marker was a useful tool in determining genetic variation of inter and intra specific of rye.

FZD7 receptor is considered as an emerging target for the treatment of Wnt-βcatenin related cancers. This transmembrane receptor is overexpressed in many cancer types like breast cancer and ovarian carcinoma, and so selective targeting of this receptor has a great therapeutic capacity. On the other hand, one of the mechanisms proposed for the anticancer effect of Atrial natriuretic peptide (ANP) that known as a heart hormone at first, is Wnt-βcatenin inhibition through an FZD dependent manner but, the molecular mechanism of this inhibition is not clear. Here, using computational methods including molecular docking and molecular dynamics simulation, also designing a cellular system that enabled us to trace Wnt-βcatenin kinetics directly, we investigated the mechanism of the peptide inhibitory potential against the pathway. Our computational results show that ANP can directly interact with FZD7 and also, its binding site on FZD7 overlap to the binding region of the Wnt carboxyl-terminal domain (Wnt-CTD). The finding of the silencing experiments demonstrates the dependency of Wnt-βcatenin signaling of the cellular system to FZD7. The decrease of βcatenin in cells treated to ANP and Wnt is also significant to compare to the control experiments. Finally, our results show that ANP is a potential scaffold to design selective peptide against FZD7.

Nerve growth factor (NGF) is a neurotrophic factor that is functional in survival, maintenance and differentiation of peripheral and central nervous system cells. This protein has three subunits that its beta subunit has main activity. According to scientific studies, it can be used as a therapeutic agent in treatment of many diseases such as peripheral neuropathy associated with diabetes, Alzheimer's disease, Parkinson's disease, skin disease and so on. Prokaryotic expression of recombinant NGF should be done in the periplasmic space because of its oxidative envoronment. It is worth noting that co-expression of cytoplasmic molecular chaperones can facilitate the secretion of the recombinant proteins to the periplasmic space and also enhance the protein solubility. In this study, the effect of cytoplasmic chaperones of GroEL / GroES, DnaK / DnaJ, GrpE, Trigger Factor (TF) on the periplasmic production of recombinant NGF protein was studied. For this purpose, β-NGF subunit was expressed in pET39b(+) expression vector simultaneously with chaperone plasmids pG-Tf2, pTf16, pGro7, pKJE7 and pG-KJE8 in E. coli DE3 strain. The results showed that in the presence of TF chaperone (pTf16 plasmid), the total protein and periplasmic production increased. Also, the DnaK/DnaJ and GroEL/GroES chaperones (pG-KJE8 plasmid) have also increased the production to some extent.; while the expression of GroEL/ GroES (pGro7) or DnaK / DnaJ (pKJE7) had no effect on protein expression. Also treatment of PC12 cell line with recombinant β-NGF showed differentiation to nerve cells which indicates that the produced protein is fully functional.

Hyaluronic acid (HA) is a natural and linear polymer that finds a wide-range of applications in medicine, cosmetics, and nutraceuticals because of excellent viscoelasticity, high moisture retention capacity, high biocompatibility and non-toxicity. HA has been recently produced in industrial scale by Streptococcal species. Streptococci are nutritionally fastidious lactic acid bacteria and cannot synthesize some amino acids. Therefore, it is necessary to study and select some commercial culture media for their growth. In this study, HA production and hyaluronidase activity of S. zooepidemicus ATCC 43079 in three culture media were investigated. Regarding the detrimental effect of this enzyme on HA amount, 6-O-Palmitoyl-L-ascorbic acid as hyaluronidase inhibitor was added to culture medium during fermentation. The effect of three variables consisted of glucose concentration, yeast extract concentration and medium pH each at 3 levels were considered and (response surface methodology (RSM) was used for statistical design of experiments to study the HA production by this strain. The results showed that maximum HA production was obtained when glucose concentration, yeast extract concentration and pH were 21.2 g L-1, 43.6 g L-1 and 6.6, respectively. Under optimum conditions, HA was produced as 370±15 mg L-1 which was ~150% more than of HA concentration in basal medium (150±10 mg L-1) and productivity reached 56.74 g L-1 h-1 that was increased 2 fold compared to central point.

The secondary metabolites of plants have been considered due to their diversity and roles in plants and human health. Grapevine is one of the plants that have secondary compounds with medicinal properties. such compounds include resveratrol which is a phenolic compound from the stilbenoid group. In order to investigate the resveratrol production under the effect of an elicitor, a CRD design with four replications using Soltani cultivar was done.

 The salicylic acid (SA) was used as an elicitor with variable concentrations including 0, 10-4, 10-5, 10-6 and was introduced into MS medium without hormones to examine it`s stress effects. At the molecular level, the effect of SA on the expression of stilbene-synthase gene was evaluated by semi-quantitative RT-PCR. In the biochemical experiment, the rate of resveratrol production was measured by HPLC.

 Stilbene-synthase expression analysis showed that SA with a concentration of 10-4 M had a positive and incremental effect on gene expression and showed a 35.48% of increase compared to the control state, and also the concentration of 10-5 M increased the gene expression 5.65% in relation with control state. In the biochemical experiment, increasing in production of resveratrol was observed in 10-4 M treatment compared to the control treatment (6.1 µg) and 10-5 M treatment (3.25 µg) did not show a significant difference with the control sample.

 SA as an elicitor and stimulant of resveratrol production could enhance the expression of stilbene-synthase gene followed by enhancing the medicinal properties of the Vitis plant.

This article has been attempting to present a novel and biocompatible nanocarrier based on chitosan biopolymer with high efficiency and low toxicity for gene delivery. At the first, 5-amino-1H-tetrazol reacted with (3-chloropropyl)trimethoxysilane and then, the obtained organosilane intermediate was used for functionalization of chitosan amino groups. The structure and chemical composition of the synthesized nanocarrier were fully characterized by FESEM, TEM, XRD, FTIR, Zeta potential and DLS analysis. At the fallowing, the MTT test was performed in order to investigation of the nanocarrier toxicity. Furthermore, Hek-293T cell line was used for investigation of the nanocarrier efficiency in gene transfection toward the cell. The loading capacity of plasmid was optimal at an N/P ratio of 3 with excellent protection of the plasmid. The results were shown that the novel nanocarrier is high efficient and has high potential to gene transfection and protection in addition of nontoxicity, biodegradability and cost effectively.