مجله پژوهش های سلولی مولکولی (زیست شناسی ایران)
سال سی و چهارم شماره 4 (زمستان 1400)

The cell migration and movement is a critical step in metastasis processes and moved from initial tumor to secondary through capillaries of lymphatic systems. Conventional methods for cell migration studies have significant limitation in study of cell heterogeneity and simulated the limited step in metastasis process which is moved cell through capillaries of lymphatic systems. Moreover, all of these systems are endpoint assays. In our study, we designed a microfluidic chip to study of cell migration, metastasis properties of cells and cell heterogeneity of cancer cells. In our designed pattern, there are several trapping shapes in main channel which the cells trapped in these positions. In addition, the narrow channels were designed in front of main channels which cell migrate through it and simulated the capillary of the lymphatic systems during metastasis process. In order to investigate the cell migration, we monitored the viability of the cells in microfluidic chip and the cell migration properties of MCF-7 and MDA-MB-231 cell lines and carried out statistical analysis on cell migration properties of both cell lines. Our results showed the microfluidic chip support cell growth on both cell lines. The theoretical and experimental results indicated the gradient of chemoattractant agent was generated in microfluidic chip. Our findings show that the mean velocity of MDA-MB-231 cells were greater than MCF-7 cells. Based on our result, the mean velocity of MCF-7 and MDA-MB-231 cell line were 18.4 µm.h-1 and 22.1 µm.h-1. ....

In normal physiological conditions, proteins are mostly spherical and water-soluble. In some cases, proteins are converted into structures called fibrils or amyloid plaques that are insoluble and pathogenic. In this study, the effects of hydroalcoholic extracts of Quercus brantii fruit-hull on the formation of hen egg-white lysozyme fibrils as well as the effect of this extract on the elimination of preformed HEWL fibrils have been investigated. For this purpose, lysozyme was incubated in presence and absence of Q. brantii extract at 60 ˚C for 15 days. The effect of extract was evaluated using CR binding assay, CD spectroscopy, THT and ANS fluorescence testing, and FE-SEM microscope.The results showed that Q. brantii extract in all tested ratios has an inhibitory effect on the formation of amyloid fibrils. Also, the extract has an accelerating effect on the defibrillation of amyloid aggregates and the greatest effect of it on defibrillation is observed in the ratio of 0.5: 1 extract to fibril. From the results obtained in this study, it can be concluded that Quercus brantii fruit-hull contains compounds that can be used as agents in the preparation of drugs for the removal of amyloid plaques in the treatment of amyloid-dependent diseases.

Nowadays heterologous expression of proteins plays a key role in biotechnology. Inclusion body formation in heterologous expression of proteins, especially expression of eukaryotic proteins in prokaryotic hosts including E. coli, is one of the most laborious challenges for researchers. High expression of protein, its specific conformation, disulfide bonds and protein charge are account for inclusion body formation. It has been reported that some inclusion bodies have biological activity but in most cases they should be renatured and soluble. Refolding processes are often not only multi-steps and time-consuming but also have low yields. In this study for the first time MO-CBP2 from Moringa Oleifera seed, successfully expressed heterologously in the E. coli but formation of inclusion bodies was a great challenge. Different methods were carried out for refolding the protein, more efficient one was Nickel sepharose column affinity chromatography with urea gradient 8-0 M that result in simultaneous refolding and purification of the protein in fewer steps. This method yields a considerable amount of pure and renatured protein and it is recommended as a convenient and efficient method.

Due to the severe limitations of fresh water in Iran, the use of low-quality saline water resources is an inevitable necessity in development of sustainable agriculture. One of the ways to deal with climate change is to use the potential of halophyte plants such as Salicornia for the economic utilization of saline soils. In this study, two genotypes of Salicornia (Qom, sensitive to salinity and Helleh, tolerant to salinity) were hydroponically treated with salinity and different concentrations of phosphorus (P) and magnesium (Mg). The aim of this study was to investigate the role of these two elements on growth factors and morphological and anatomical characteristics in Salicornia under the salinity treatments. The results showed that at 0 mM salinity, P could stimulate growth in both genotypes. But at 200- and 800- Mm salinity, had a negative effect on growth parameters. Increasing the concentration of Mg at all three salinity levels(0, 200, 800mmol) improved growth in both genotypes. Also, the lack of P in 200-and 800-mM salinities affected the plant anatomy and caused lignification areas in the endoderm and increased diameter in the cell wall of the xylem. In addition, a 3-fold concentration of Mg at 800 mM salinity severely deformed and plasmolyzed epidermal and parenchymal cells. Overall, these studies showed that a one-fold treatment of Mg and P provides the best conditions for the growth of Salicornia.

Ovarian cancer is the seventh most common cancer in women and the fifth most common type of cancer in Iranian women. Salvia officinalis is one of the most important genus of dark mint, most species of which have medicinal properties and are widely used in traditional medicine. The aim of the present study was to evaluate the essential oil of salvia on the rate of proliferation and apoptosis of ovarian cancer cells. The salvia officinalis plant was prepared from the Iranian Biological Research Center and the oil obtained by Clevenger apparatus and its components identified by gas chromatography. Ovarian cancer cells of SKOV3 type were treated with different concentrations of (100, 200, 300, 400, 500 and 600 µg/ml) and their viability assessed by trypan blue absorption and MTT at three time periods of 24, 48 and 72 hours and the rate of induction of cellular apoptosis was assessed by flow cytometry. Cell survival decreased with increasing concentration of essential oils in a dose-dependent manner. The IC50 level of Salvia officinalis essential oil for 48 hours was 300 μg/ml. Induction of apoptosis was also dose-dependent, and apoptosis index increased significantly at a dose of 600 μg / ml. Based on the results, it can be concluded that Salvia officinalis essential oils can reduce the biological ability and increase the apoptosis of SKOV3 ovarian cancer cell cancer.

Compounds used in pharmaceutical, nutritional and cosmetic should be non-toxic. One of the toxicity tests is performed using Artemia spp. as sensitive organisms to various toxins. In these tests, various characteristics of these larvae are investigated. One of the most commonly used factors is evaluation of the survival and death rate of larvae. Detection of live and dead larvae is based on the motility of larvae. However, the rapid movement of larvae makes it difficult to count, and thus the reliability of the test decreases. This study aimed to develop a program for the automatic differentiation of live and dead larvae and their counting in a short time and with high precision. In this regard, a program was designed using image processing rotes to distinguish larvae in MATLAB software. Motility of larvae was detected by image subtraction. The program receives images sent from the camera at two time range (20 seconds) at the end of incubation period and then processes. At each interval, the background is calculated for the same interval. This will cause larvae that have died so far to be added to the background and not counted in the detection of live larvae. Eventually, the program could accurately count live and dead larvae. There is no significant difference between findings from the program and the findings provided by the expert user (P>0.05). Benefits of this improved test with programming are lack of need to aseptic conditions, costly materials and equipment and user presence as well as reduced errors.

Atropa Belladonna L. is the most important medicinal plant of Solanaceae family, due to important commercial sources of atropine and scopolamine as tropane alkaloids. Applications of hairy root cultures are advised because of their special properties such as fast growth, short doubling time, ease of maintenance, and ability to synthesize and continuous production of a range of chemical compounds. Effects of ozone depletion and UV irradiation have been studied by most of the investigators. In the present study, the effect of different levels of UV-B irradiation was studied on atropine production and activity of some antioxidant enzymes. UV-B irradiation during 3 days was treated for 4, 8 and 12 min that was equal to 13, 26 and 39 Kj/m2 respectively. Results showed that increase of UV-B lending to decrease hairy roots dry weight. This irradiation increased atropine content in comparison of control treat and the highest was recorded in 26 Kj/m2. The activity of antioxidant enzymes including catalase, guaiacol peroxidase, and ascorbate peroxidase was also elevated in treated hairy roots rather than of the control at 0.01% of probability. Based on obtained results in this work UV-B irradiation can be considered as one of the useful methods for Atropine and antioxidant enzymes in A. belladonna hairy roots.

In this study, we developed a single specific primer PCR (SSP-PCR) method for specific identification of Shigella genus with the use of ribosomal RNA gene. Regarding high copy number and stability in cells, rRNA is one of the efficient targets to detect cells exclusively. 16S rRNA gene is considered as a standard biomarker for classifying and identifying prokaryotic cells. With respect to high sequence homology among 16S rRNA gene and shortness of specific heterogenic region in this gene, SSP-PCR is used to facilitate the specific identification of 16S rRNA gene in this study. In this method, only one specific primer is designed for amplification of target genes. 16S rRNA gene of different species of Shigella genus was aligned with 57 other related bacteria and then a single specific primer was designed in the V6 variable region. The amplicon was analyzed on a two-dimension agarose gel electrophoresis. The 263 bp band was considered as positive result in the tested samples and was observed for all species of Shigella (Sh. flexneri, Sh. boydii, Sh. dysenteriae, Sh. sonnei), whereas other species remained unidentified. The specificity of the assay was evaluated to be complete. Combination of targeting 16S rRNA and using SSP-PCR method, is an efficacious method for specific detection of bacteria in a minimum required time with a great ease. Among the applications are namely the early-phase diagnosis of bacterial infections in acute emergency conditions such as natural disasters and countries’ defense industry in the form of diagnostic kits for clinical laboratories.

Citric acid is one the most commonly used organic acid in the food and pharmacntical which is produced mainly by submerged or surface fermentation by Aspergillus niger. As the most important of commereial producer. Recent works showed successful citric acid production through solied state fermentation. Under our experimental conditions, four important factor each one in four levels including substracte type (wheat straw, rice straw, rice bran, peanut skin) and pH (4,5,6,7) substrate treatment tempretures (45,60,75,90,.c) and substracte treatment time (30,60,90,120min) on citric acid production by Aspergillus niger were assayed. Maximum amount of citric acid production was pH =5, treatment time and temperature of 60(min) and 60 (.c) respectively also the wheat straw was the best substrate for citric acid production. The results of this study showed that Aspergillus niger is a suitable microorganism for citric acid production and wheat straw is a good base for economical production of citric acid due to its low price. All of four understudied factors had significant effect on citric acid production in solid state fermentation (P < 0.05).

Gene expression is regulated with miRNAs, that are non-coding small RNA molecules and genetic variations such as miRNA genes SNPs, which are molecular biomarkers in the prediction of cancers risk. One of miRNAs that is associated with BRCA1 / BRCA2 genes, is miR146a and related with many cancers, including breast cancer. This research aim was association study between prevalence of rs2910164 SNP in miR146a gene with breast cancer in Iranian female population.

This study was performed on 50 patient and 50 control blood samples. DNA was extracted of samples. Then for genotyping was used Tetra Aarms PCR and results was statistically analyzed.Results- Frequency of genotypes for GG, GC and CC in control groups were 30%, 66% and 4%, and for patient groups was 46% , 52% and 2%, respectively. No significant difference was observed between frequency of dominant and recessive allels in control and patients gorups (P > 0.05).

In this study results, there was not significant difference between G and C alleles in the control and case groups. Therefore, could be indicated that there was not association between of miR146a gene rs2910164 polymorphism and breast cancer in the studied Iranian females population. So, further studies with larger sample size are required to support finding of this research.

Mutations in the FLT3 gene have already been reported in leukemia especially acute myeloid leukemia (AML). The diagnostic value of such mutations in leukemia is in the management of high-risk patients and the targeted therapy of diseases. Recently, the researchers provide this important molecular marker in diagnosis of AML patients. Our aim in this study is to analysis the prevalence of the FLT3 gene mutations as a valuable molecular marker which could be used in the diagnosis and treatment of AML patients with normal karyotype and prevalence survey of mutation in Northeast Iran.

In this study, fresh bone marrow samples from 83 patients with untreated acute myeloid leukemia which have normal karyotype were evaluated for the presence of two mutations in the FLT3 gene by PCR and PCR-RFLP (to detect FLT3-ITD mutation).

The prevalence of mutation in FLT3 gene was 27.7% between AML patients. Of all patients, 19.3% showed FLT3-ITD mutation and 8.4% FLT3-TKD mutation. Most patients with mutations were in the M2 subgroup. Regardless of age and sex, there was a significant increase in white blood cell count among patients carrying FLT3 mutations (p< 0.05).

The use of mutations such as FLT3-ITD and FLT3-TKD as biomarkers are highly effective in generating a more comprehensive picture of the pathophysiological processes involved in the diseases and can significantly improve disease diagnosis, follow-up and monitoring of patients, so physicians will make more targeted treatment decisions.