Halimeh Hassanzadeh; Ahmad Reza Bahrami; Hamidreza Bidkhori; Maryam M. Matin
Abstract
Transplantation of mesenchymal stem cells (MSCs) is a promising strategy in regenerative medicine. These cells can differentiate into chondrocytes, fibroblasts, or osteoblasts, essential components in bone healing. Dysregulated inflammation, resulting from a decreased or augmented immune response, can ...
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Transplantation of mesenchymal stem cells (MSCs) is a promising strategy in regenerative medicine. These cells can differentiate into chondrocytes, fibroblasts, or osteoblasts, essential components in bone healing. 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 an 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; 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.
Maryam M. Matin; David P. Hornby
Abstract
We have developed a rapid, quantitative method for analysing the outcome of the first strand synthesis step in cDNA library preparation, yield and molecular weight range of the final cDNA products are determined after size fractionation. This method involves conventional cDNA library ...
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We have developed a rapid, quantitative method for analysing the outcome of the first strand synthesis step in cDNA library preparation, yield and molecular weight range of the final cDNA products are determined after size fractionation. This method involves conventional cDNA library construction including all enzymatic steps usually required, but replaces radioactive labelling of nucleic acids with fluorescence detection. The separation and quantification steps all involve ion-pair reversed-phase high performance liquid chromatography (IP RP HPLC). This quantitative method replaces the use of autoradiography and size exclusion chromatography with combined ion-pair reversed-phase high performance liquid chromatography and in line fluorescence detection. The result of this approach is combination of speed with the generation of reproducible, high quality cDNA libraries.
Zahra Jomehzadeh; Farhang Haddad; Maryam M. Matin; Shokouhozaman Soleymanifard
Abstract
There are several studies suggesting the role of aneuploidy in tumor formation. Aneuploid cells are different from normal ones in term of gene expression and proteome. Cells with different amount and kind of proteins might act differently to external stimuli, including ionizing irradiation. ...
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There are several studies suggesting the role of aneuploidy in tumor formation. Aneuploid cells are different from normal ones in term of gene expression and proteome. Cells with different amount and kind of proteins might act differently to external stimuli, including ionizing irradiation. Currently, radiotherapy is one of the main methods in fight against cancer, therefore, it is important to understand the response of the aneuploidy-tumor cells to irradiation. To investigate the chromosomal effect of gamma irradiation on aneuploid cells, L929 cells were treated with 1.5 ng.ml-1 of vinblastine to induce aneuploidy. Vinblastine-treated cells were left to recover for 72 h and irradiated with 1 Gy of gamma radiation. Induced chromosomal damages were investigated using micronucleus (Mn) assay. Data showed that vinblastine and gamma irradiation both were able to significantly increase micronucleated-binucleated cells (MnBi) frequency. However, 1 Gy gamma irradiation of the cells after 72 h of vinblastine treatment led to the lower frequency of MnBi compared to irradiated cells. Results of this study suggest that vinblastine treatment of cells before irradiation not only did not sensitize the cells to radiation-induced chromosomal abnormalities, but also had radio-protective effect for these cells. This result could be useful in planning cancer therapy regimes.
Farahnaz Molavi; Jamshid Darvish; Farhang Haddad; Maryam M. Matin
Abstract
Cytotaxonomy is a branch of cytogenetics, devoted to the comparative study of karyological features for systematic and evolutionary purposes. Surely, awareness of chromosomal characters increases our knowledge in different fields of studies. In this study, cytogenetic analyses were performed in 92 Mus ...
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Cytotaxonomy is a branch of cytogenetics, devoted to the comparative study of karyological features for systematic and evolutionary purposes. Surely, awareness of chromosomal characters increases our knowledge in different fields of studies. In this study, cytogenetic analyses were performed in 92 Mus musculus specimens from 26 localities in Iran. Cytogenetic characteristics of the house mouse, Mus musculus, in Iran show that the chromosome number is 2n=40 and the arm number is NF=40. The karyotyping results indicated the presence of 20 Acrocentric (A) chromosome pairs. The L/S (r ratio) was between 2.0621 and 4.5862. The length of shortest chromosome, length of longest chromosome and mean of chromosomal length in different populations were between 2-3.58, 6.07-7.01 and 3.43-5.05 (μm), respectively. The results showed two distinct karyotypic formulae, namely cytotype B and cytotype C. Asymmetry indexes (AI, DI, As%, A, A2, A1 and Syi%) in all population except Birjand and Khash showed symmetry in chromosomes. In clustering methods using the matrix of symmetrical indexes similarities, four clusters were revealed, one for specimens of central and east of Iran, the second cluster for specimens from south and west of Iran, the third cluster was related to the eight specimens of Birjand and finally, the fourth cluster for two specimens of Khash locality
Habib Rezanejad; Farhang Haddad; Zahra Soheila Soheili; Maryam M. Matin; Shahram Samiei; Sepideh Zununi Vahed
Abstract
Glaucoma remains one of the major causes of blindness in today's world. The progressive field of stem cell proposes an exciting potential for discovering novel therapies. Here, we report the development of an easy and high throughput method for differentiation of retinal ganglion cells (RGC) and bipolar ...
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Glaucoma remains one of the major causes of blindness in today's world. The progressive field of stem cell proposes an exciting potential for discovering novel therapies. Here, we report the development of an easy and high throughput method for differentiation of retinal ganglion cells (RGC) and bipolar cells from human adipose tissue-derived mesenchymal stem cells (hADSCs) using PAX6 (+5a) gene expression, a master gene in development of the vertebrate visual system. HADSCs was isolated from fat tissues and confirmed by their surface markers and differentiation potential into adipocytes and osteocytes lineages. Then, the coding region of human PAX6 (+5a) gene was cloned and lentiviral particles were produced. HADSCs differentiation was characterized by morphological characteristics, qRT-PCR and immunocytochemistry (ICC). The hADSCs were isolated successfully with high yield and purity (99%). After 30 hours post transduction by pLEX-pax6- pur lentiviral vectors in fibronectin supplemented medium, cells gradually showed the characteristic morphology of neuronal cells. QRT- PCR and ICC confirmed deriving of mainly RGC and marginally bipolar cells. The current investigation demonstrates the feasibility of differentiation of RGCs and bipolar cells from hADSCs using expression of PAX6 (+5a) in the medium supplemented by fibronectin.
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.
Maryam M. Matin
Abstract
Cancer remains a major cause of death worldwide. Huge research and identification of several markers have resulted in better understanding of its mechanism. Researches focusing on cancer stem cells and their role in metastasis will help the scientific community to propose the therapeutic approaches for ...
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Cancer remains a major cause of death worldwide. Huge research and identification of several markers have resulted in better understanding of its mechanism. Researches focusing on cancer stem cells and their role in metastasis will help the scientific community to propose the therapeutic approaches for treatment of this monstrous disease. Interest of governmental agencies and inter-communications of molecular biologists with clinicians can boost the new ideas in identification and characterizations of cancer stem cells. It will also help to elucidate their roles in tumor progression and hopefully would result in better ways to reduce the mortality related to cancer.
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.
