Halimeh Hassanzadeh; Ahmad Reza Bahrami; Hamidreza Bidkhori; Maryam M. Matin
Abstract
Transplantation of mesenchymal stem cells (MSCs) is considered a promising strategy in regenerative medicine. These cells can differentiate into chondrocytes, fibroblasts, or osteoblasts, as essential components in the bone healing process. Dysregulated inflammation, resulting from a decreased or augmented ...
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Transplantation of mesenchymal stem cells (MSCs) is considered a promising strategy in regenerative medicine. These cells can differentiate into chondrocytes, fibroblasts, or osteoblasts, as essential components in the bone healing process. Dysregulated inflammation, resulting from a decreased or augmented immune response, can suppress bone healing. To overcome this problem, different strategies have been applied to improve the anti-inflammatory and immunomodulatory potencies of MSCs. Several studies have explored the potential of using small molecules to enhance the process of bone formation and regeneration. In addition to the proven safety and efficacy of lithium in managing bipolar disorder over many years, it has been reported in several studies that it could potentially contribute to an increase in bone mass. Some have focused on the role of lithium chloride (LiCl) in activating the WNT/β-Catenin pathway, which is involved in the differentiation of MSCs into osteoblasts. In this study, we evaluated the ability of adipose-derived mesenchymal stem cells (Ad-MSCs) treated with LiCl to differentiate into bone cells. To assess osteogenesis, mineralization was evaluated in cells cultured in osteogenic induction medium. In addition to checking the expression of genes related to bone formation, we also investigated the expression of several genes related to immunomodulation at the mRNA level. We observed that LiCl enhanced the osteogenesis of Ad-MSCs, as evidenced by an increase in mineralization and the enhanced expression of osteogenic markers. Moreover, the expression of cytokines, which promote the anti-inflammatory behavior of these cells, was augmented. These findings could potentially be clinically relevant to improving conditions associated with bone loss, such as osteopenia and osteoporosis.
Seyed Navid Goftari; Ahmad Reza Bahrami; Maryam M. Matin
Abstract
Esophageal cancer is one of the most aggressive gastrointestinal malignancies, and esophageal squamous cell carcinoma (ESCC) is the most prevalent esophagus neoplastic disease with high mortality rates in some Asian countries. Nonetheless, the etiology of ESCC continues to be vaguely comprehended, and ...
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Esophageal cancer is one of the most aggressive gastrointestinal malignancies, and esophageal squamous cell carcinoma (ESCC) is the most prevalent esophagus neoplastic disease with high mortality rates in some Asian countries. Nonetheless, the etiology of ESCC continues to be vaguely comprehended, and the role of long noncoding RNAs in different clinical stages of this type of malignancy remains to be clarified. Here, we aimed to investigate the crucial genes corresponding to various clinical stages of ESCC, determine the hub lncRNAs in these stages, and predict patients’ overall survival time. In the current study, the cancer genome atlas (TCGA) RNA-seq public data was analyzed in order to discover novel biomarkers or therapeutic targets implicated in the progression of ESCC. Stage-related genes were analyzed, the protein-protein interaction network for any stage was constructed and the top 5 genes with the most Maximal Clique Centrality score in each network were selected as the hub mRNAs. LncRNAs interacting with each stage hub mRNA were also determined as stage-related hub lncRNAs. Gene set enrichment analysis on stage-associated modules was also carried out. Finally, Cox regression analysis was performed to assess the prognostic significance of identified hub lncRNAs in the survival of patients with ESCC. Finally, hub mRNAs and hub lncRNAs associated with ESCC progression were identified, which may have implications as biomarkers and targets for therapeutic interventions. Six lncRNAs, including AC013391.2, AC104088.1, AC026341.3, AL139023.1, AL583808.1, and LINC01707 were also identified to be significantly correlated with ESCC patients’ overall survival time, which could be potential predictors for the survival rate of patients, however, more research is required in order to confirm the results experimentally.
Najmeh Sodagar; Ahmad Reza Bahrami; Maryam Moghaddam Matin
Abstract
Salmonella is a gram-negative bacillus that lives in the intestinal tract of human and animals and causes diarrhea. Salmonella could be found in undercooked products of poultry with no impact on the taste, smell, or appearance. Since poultry eggs and meat might be sources of Salmonella and pose a hazard ...
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Salmonella is a gram-negative bacillus that lives in the intestinal tract of human and animals and causes diarrhea. Salmonella could be found in undercooked products of poultry with no impact on the taste, smell, or appearance. Since poultry eggs and meat might be sources of Salmonella and pose a hazard to public health, it is important to accurately detect Salmonella infection. In this regard, the present study aimed to develop a rapid and sensitive method for the diagnosis of Salmonella spp. in samples from the poultry industry. To do so, the sensitivity of S. enterica serotype Enteritidis detection was assessed with ten-fold serial dilutions in peptone water to give suspensions containing 100 to 105 CFU/mL. For artificial inoculation, skin samples were sequentially inoculated with the serial dilutions, while a control sample was included to ensure that the skin was not naturally contaminated with Salmonella. 53 commercial chicken skin samples were obtained from different local shops. Then, DNA was extracted from all samples, and the quality of extracted DNAs was checked by spectrophotometry and confirmed by agarose gel electrophoresis. For PCR, a pair of oligonucleotide primers, INVA, was designed to amplify the invA gene. Results revealed a band of 796 bp in samples artificially contaminated with S. Enteritidis. Likewise, the 796 bp band was detected in 38 samples (71%) with deferent intensities, which presented different amounts of contamination. Accordingly, the present study provided a valuable method for the detection and control of Salmonella infection in the poultry industry, since results would be available in less time than with the conventional cultural method.
Azadeh Haghighitalab; Mahboubeh Kazemi Noughabi; Shima Minaee; Ahmad Amin; Ahmad Reza Bahrami
Abstract
Acute myocardial infarction (MI) describes as an irreversible death of heart muscle which is initiated by a shortage of myocardium oxygen supply and accompanies by a complex of pro- and anti-inflammatory events. During the last decades, innate and adaptive immune responses are considered ...
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Acute myocardial infarction (MI) describes as an irreversible death of heart muscle which is initiated by a shortage of myocardium oxygen supply and accompanies by a complex of pro- and anti-inflammatory events. During the last decades, innate and adaptive immune responses are considered more serious for controlling myocardial infarction. As, it was confirmed that deregulated immune system which triggers excessive local and systemic inflammatory events is responsible for serious adverse effects associated with acute MI. Bone marrow activation, spleen monocytopoiesis, a remarkable increase of circulating cytokines and adhesion molecules, in addition to elevated levels of active peripheral leukocytes and platelets are playing significant roles in determining the clinical outcome of patients with MI. The previous experience demonstrated the failure of traditional harsh anti-inflammatory strategies. High mortality rate and poor quality of life observed for survivors of MI despite current progress in the field highlight the urgent need for such interdisciplinary studies in the context of molecular cardiology. Hence, unraveling the cellular and molecular events which are involved in the management of inflammatory responses post-MI is of special focus. The concept of immune regulation after myocardial infarction is not new, but our perception for dealing with the challenge has been changed during the last decades with gaining more in-depth molecular/immunological knowledge. It seems that fine-tuning the interplay between innate and adaptive immune responses and regulating their cross-talk should be in special focus to establish effective therapeutic strategies.
Azadeh Haghighitalab; Maryam M. Matin; Fatemeh Khakrah; Ahmad Asoodeh; Ahmad Reza Bahrami
Abstract
Despite the prominent therapeutic potentials of stem cells, their use in cell therapy has been challenged with some unreproducible and inconsistent outcomes in addition to the risk of rejection and tumorigenesis. Gaining novel insights to the importance of the conditioned medium, ...
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Despite the prominent therapeutic potentials of stem cells, their use in cell therapy has been challenged with some unreproducible and inconsistent outcomes in addition to the risk of rejection and tumorigenesis. Gaining novel insights to the importance of the conditioned medium, secretory factors and extracellular vesicles as the functional components of the cultured stem cells, suggested the idea of substituting the cells with their cell-free counterparts. Biological properties of these products are influenced by the cues received from their microenvironment. Hence, providing optimal and fully defined culture conditions is essential for their preparation. Fetal bovine serum (FBS), one of the most routine supplements of cell culture, is enriched by endogenous extracellular vesicles (EVs). These EVs will affect the yield, purity and functional features of the cell-free products. Here, we endeavored to examine and compare three different methods including ultrasonication, ultrafiltration and polymer-based precipitation, to deplete EVs from FBS. We chose easy to perform and fast methods with the capacity for high-throughput applications. Based on our observations, although all examined methods were able to deplete EVs from FBS to some extent, polymer-based precipitation could be considered as the method of choice with minimal consequences on the biological requirements of FBS to support cell growth and characteristics. Due to similarities between FBS and some other biological solutions, this strategy would be suitable for EV-depletion from other liquids with high concentrations of proteins and nutrients. Moreover, it could be applied for preparation of optimal culture conditions for nanoparticle applications.
Maria Beihaghi; Ahmad Reza Bahrami; Abdolreza Bagheri; Mohammad Zare Mehrjerdi
Abstract
Studies on the genes contributing to the seed filling in chickpea and its protein content might be valuable in engineering plants with seeds of a higher nutritional value. A gene of interest is phosphoenolpyruvate carboxykinase (pepck), encoding a protein with a substantial role in the gluconeogenesis ...
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Studies on the genes contributing to the seed filling in chickpea and its protein content might be valuable in engineering plants with seeds of a higher nutritional value. A gene of interest is phosphoenolpyruvate carboxykinase (pepck), encoding a protein with a substantial role in the gluconeogenesis pathway. In the present study, the protein content (percentage) was measured in a number of cultivated chickpea genotypes, followed by comparison of the expression levels of pepck gene at different stages of seed filling in some of the genotypes. This study aims at revealing the relation between pepck transcript level with protein content of chickpea seeds which might, in the longer term, end at protein quality improvement of the crop through the gene manipulation procedures. The results which were verified by Real Time-PCR and Western blot techniques in four genotypes of plant, showed that, the amount of pepck expression was significantly higher at the stage of seed fillingthan at other stages in all of genotypes and the lowest levels of expression belonged to flowering and seed formation and the PEPCK protein level was higher in the high protein genotypes compared to the low protein genotypes.
Seyed Ali Mortazavi; Ahmad Reza Bahrami; Balal Sadeghi; Maryam Moghaddam Matin
Abstract
In present study, a SYBR green based real time PCR assay was developed for specific detection and quantification of Bacillus subtilis in dough used for bread making. New primer pairs were designed to amplify a 212 base pair fragment of the aprE gene. Specificity of these primer pairs was confirmed with ...
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In present study, a SYBR green based real time PCR assay was developed for specific detection and quantification of Bacillus subtilis in dough used for bread making. New primer pairs were designed to amplify a 212 base pair fragment of the aprE gene. Specificity of these primer pairs was confirmed with conventional and real time PCR methods. Standard curves constructed using the threshold cycle (CT) versus copy numbers of B. subtilis showed good linearity for reference standards of cloned insert (R2=0.999, slope=-3.035) and also induced contaminated dough (R2=0.988, slope=-3.142), and the melting temperature (Tm=82.2 oC) was consistently specific for the amplicon. Limits of detection were 200 and 2000 colony forming units (CFUs) per ml or g of these samples, respectively. This real time PCR offers a fast tool with high sensitivity and specificity for detection and quantification of this rope-forming pathogen in dough used for bread making.
Hassan Monhemia; Mohammad Reza Hosseindokht; Mohammareza Bozorgmehr; Ahmad Reza Bahrami
Abstract
Signaling pathways are not isolated from their surroundings. They are also intervened by other signaling pathways known as “crosstalk mechanism”. One of the most important crosstalk mechanisms is the insulinEGF network. Although insulin and epidermal growth factor (EGF) networks have some complexity ...
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Signaling pathways are not isolated from their surroundings. They are also intervened by other signaling pathways known as “crosstalk mechanism”. One of the most important crosstalk mechanisms is the insulinEGF network. Although insulin and epidermal growth factor (EGF) networks have some complexity in their isolated forms, their complexities will grow in the crosstalk network. In this study, we used the analytical tools of the systems biology workbench for elucidating some ambiguities of the insulin-EGF crosstalk. Based on sensitivity analysis, we reconstructed an elucidated model with 51 chemical reactions in comparison with the previous model with 111 chemical reactions. Interestingly, this reduced model reproduces the results of the original model in synergy conditions. We noticed two controlling pathways with direct participation of phosphorylated insulin and EGF receptors that involve Insulin Receptor Substrate (IRS) and Src kinase modules. Also, insulin pathway by producing phosphatidylinositol-3, 4, 5-triphosphate (PIP3), and EGF pathway by activation of GAB1, control the downstream events and lead to potentialities in the mitogenic signal. Surprisingly, Shc and phosphatase SHP2-dependent reactions have no significant roles in the synergy conditions and are not involved in the reduced model. Regarding sensitivity analysis, all Ras/ERK cascade reactions are crucial for signal transduction and were kept in the reduced model.
Eisa B. Kohan; Mohammad B. Bagherieh-Najjar; Mahnaz Aghdasi; Ahmad Reza Bahrami
Abstract
Environmental stresses affect plant growth and cause losses worth hundreds of million dollars of agricultural industry each year. Many genes induced in response to environmental stresses. The DREB1A gene is a stress-inducible transcription factor that its overexpression increased tolerance to environmental ...
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Environmental stresses affect plant growth and cause losses worth hundreds of million dollars of agricultural industry each year. Many genes induced in response to environmental stresses. The DREB1A gene is a stress-inducible transcription factor that its overexpression increased tolerance to environmental stress in transgenic plants. DNA was extracted from Arabidopsis thaliana var. Col.0 plants and DREB1A gene amplified by particular primers. After purification, PCR products were cloned into pGEMT-EASY vector and transferred into E. coli strain DH5α competent cells. Blue/white colonies were analyzed and present of the DREB1A gene revealed by restriction analysis and sequencing test. A 668-bp XbaI/BamHI digested fragment of DREB1A gene from pGEMT::DREB1A construct cloned in pBI121 plasmid. The recombinant plasmids were transferred into Agrobacterium tumefaciens cells strain of LBA4404 and transformed cells were cultured on selective medium that supplied with kanamycin and rifampycin. As a result of gel electrophoresis of Agrobacterium colonies-PCR product, band existing on expected size show that the DREB1A gene was cloned in to pBI121 binary vector. At time, Agrobacterium cells containing of pBI121::DREB1A construct are using for product of environmental stress tolerant plants.
Samaneh Attaran Dowom; Ahmad Reza Bahrami; Parwaneh Abrishamchi; Morvarid Saeinasab
Abstract
Essential oils, with plant origin, have been of special attention in cancer research during recent years. Despite many reports on cytotoxic effects of plants from genus salvia, the potential application of their extracts in cancer therapy remains to be assessed in more precise and detailed examinations ...
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Essential oils, with plant origin, have been of special attention in cancer research during recent years. Despite many reports on cytotoxic effects of plants from genus salvia, the potential application of their extracts in cancer therapy remains to be assessed in more precise and detailed examinations on the main cause of such effects. In this research, the cytotoxic effect and anticancer activity of essential oils from S. leriifolia on human Transitional Cell Carcinomaa (TCC) were studied in vitro. The antiproliferative activity of essential oils on TCC and L929 (control) cells was determined by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay, by which the mitochondrial dehydrogenase enzyme activity is assessed based on reduction of the MTT to purple. The amount of essential oils to induce 50% of cells to die, designated as IC50, was determined by repeated experiments and application of different doses of the essence. The established IC50 on TCC cells for the essences extracted in two different years of 2006 and 2008 and from two locations of Bajestan and Neyshabour was respectively as: 466 and 250 μg/ml, and 233 and 212 μg/ml. S. leriifolia essential oil did not show any detectable effect on L929 cells in this range of concentration. S. leriifolia essential oil has inhibitory effects on the growth of both TCC and normal L929 cell lines, although the effective concentrations were significantly different in these cell lines. This effect was dose dependent.
Maryam Moghaddam Matin; Morvarid Saeinasab; Saeideh Nakhaei-Rad; Mahdi Mirahmadi; Nasser Mahdavi Shahri; Mahmoud Mahmoudi; Ahmad Reza Bahrami
Abstract
Abstract
Regeneration is a biological phenomenon, which takes place via two main mechanisms: first,
dedifferentiation of mature cells followed by their differentiation into functional new cells and second,
activation of endogenous somatic stem cells for regeneration of damaged or lost tissues. One ...
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Abstract
Regeneration is a biological phenomenon, which takes place via two main mechanisms: first,
dedifferentiation of mature cells followed by their differentiation into functional new cells and second,
activation of endogenous somatic stem cells for regeneration of damaged or lost tissues. One of the best
examples of healing process in mammals is the regeneration of damaged pinna in rabbits by blastema tissue.
The aim of present study was to investigate culture requirements, proliferative properties and expression of
some stemness factors in cells derived from regenerating blastema tissue obtained from rabbit pinna in vitro.
The regenerating tissues were obtained from male New Zealand white rabbits by double punching of the pinna
and cell culture conditions were set to derive and enrich the self renewing cells for further characterisation. The
cells were subjected to survival and growth examinations in vitro, and expression of several stemness factors
was studied in these cells using reverse transcription polymerase chain reaction (RT-PCR). Results revealed
that the derived cells are rather immortal, as they have been growing for more than 120 passages in culture up
until this report. Furthermore, RT-PCR and flow cytometry analyses showed that these cells express a number
of stemness related genes including Oct4 and Sox2. In conclusion, in this study, stem like cells were derived
from blastema tissue of rabbit ears for the first time, showing great self renewing capacity, which provides a
suitable in vitro model for regeneration studies. Moreover, they could be considered as a good source of stem
like cells for future applications.
Ahmad Reza Bahrami; Maria Beihaghi; Abdolreza Bagheri; Richard Leegood; Mehdi Ghabooli; Jafar Zolala; Farajollah Shahriari
Abstract
Abstract
Phosphoenolpyruvate Carboxykinase, encoded by the pepck gene, plays an important role in
gluconeogenesis. It also seems to be important in metabolism of nitrogenous compounds in developing seeds of
legumes, including amides and ureides which are then transformed into amino acids, necessary ...
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Abstract
Phosphoenolpyruvate Carboxykinase, encoded by the pepck gene, plays an important role in
gluconeogenesis. It also seems to be important in metabolism of nitrogenous compounds in developing seeds of
legumes, including amides and ureides which are then transformed into amino acids, necessary for the
synthesis of storage proteins. In this research, pepck gene expression in mRNA level, in different genotypes of
chickpea (Cicer arietinum L.), was determined. Two low protein genotypes (MCC291 and MCC373) and two
high protein genotypes (MCC458 and MCC053) out of 20 chickpea genotypes were selected. Total RNA were
extracted through different stages of seed development, and the expression of the pepck gene was estimated by
semi-quantitative RT-PCR. The results of the RT-PCR showed that two isoforms of this gene are expressed in
high protein genotypes, whereas in the low protein genotypes, the expression of these isoforms was not
obvious. Also this method showed a differential expression of pepck gene in different stages of flowering and
seed development. pepck gene is expressed in higher levels during the sheet formation and developing seeds
compared to the flowering and seed formation stages. Probably, the differential expression of pepck gene is
related to its possible role in metabolism of seed components, particularly in determination of the protein
content of chickpea seeds.
