Journal of Molecular and Cellular Researches
Year:2018 | Volume:31 | Issue:3 |Papers Count:11

Black fungi are considered as one of the original inhabitants of arid and semi-arid areas because of high ability to tolerate extreme environments. During the growth procedures, they alter the bedrocks. The present study was carried out to investigate the impacts of black rock inhabiting fungi on the tomb of Cyrus the Great bedrock. A preliminary monitoring of the tomb indicated to the presence of black fungi. Different samples of the tomb and Sivand quarry were prepared for qualitative and quantitative evaluations. The samples were evaluated with thin section preparations and Periodic Acid Schiff staining (PAS) and the results were analyzed by ImageJ software. Scanning electron microscopy technique (SEM) was used to confirm the results. The results show deep penetration of fungal hyphae into the bedrock and physico-chemical alteration of the substrate. According to the results, it seems that these microorganisms should be considered during the implementation of conservation and restoration programs.

The complement system is one of the main components of the immune system in many organisms. Existence of this system has been confirmed in numerous organisms, but there is no information about the mechanisms regulating the expression of it in some of them. In this study, we are focused on the upstream element of salmon fish C3 gene. Different tissues of the fish were prepared and DNA was extracted with a modified phenol-chloroform method. Target sequence was amplified using unclassical PCR method by using of designed degenerate primers and PCR product was purified and sequenced and finally confirmed sequence was recorded in Genomic Bank Database. In the next step, full anatomy of obtained sequence was examined for the presence of regulatory elements by application of different servers and software. Salmon fish C3 gene promoter region sequence was recorded in NCBI genomic data bank with access number JQ799884. 1. Presence of conserved TATA box and abaA, C/EBP, CTCF, IL-6, GR and PPAR transcription factors responsive elements were confirmed, however responsive element sequence for factors such as Sp1, CRE, ERE were not detected. Our results can confirm the existence of an evolutionary well-developed regulatory system on the complement system in bony fishes and support ancestral nature of complement system genes. This study is the first detailed report on the regulatory elements controlling gene expression of the C3 gene and may suggest involvement of complement derivatives in the early embryonic development steps that have been reported in some study.

Structural mf developmental characters of reproductive organs and seed in Ebenus stellata Boiss., which is the only species of Ebenus L. in Iran, were investigated for the first time using bright field, polarizing and fluorescence microscopy with different staining techniques. In this plant, the ovary has one carpel, one ovule, dense, long villi with papillae and no peduncle. The ovule is anatropous, crassinucellate, bitegmic and has endothelium. Meiosis of megasporocyte results in a T-shaped tetrad and then chalazal megaspore develops into an embryo sac with the pattern of Polygonum type. The polar nuclei are close to the egg apparatus and even after egg fertilization remain separate from each other. In embryo sac of this plant specialized tissues like hypostase, postament and a cup-like operculum are formed. In seed, a layer of macrosclereid radial cells, double palisade layer and a tracheid bar which is surrounded by parenchymal cells that contain phenolic compounds, are present. In male gametophyte, the anthers are tetrasporangiate and their wall development follows the dicotyledonous type and consists of four layers: epidermis, endothecium, one middle layer and uninucleate secretory tapetum. Microspore development is simultaneous in sporangia of an anther and arrangement of microspores is mostly tetrahedral in callosic wall. Fibrous thickenings are developed in the endothecium when shed. Mature pollen grains are ellipsoidal, tricolpate and two-celled.

The poppy (Papaver somniferum L. ) is one of the oldest medicinal plant in the world. It remains the only commercial source for the important narcotic analgesics and semisynthetic derivatives such as oxycodone and naltrexone. Several other benzylisoquinoline alkaloids with potent pharmacological properties including papaverine, noscapine and sanguinarine also produce with this plant. BBE1 is one of key genes in biosynthesis of benzylisoquinoline alkaloids and catalyzes first step in production of noscapine and sanguinarine alkaloids. In the present study, the full length CDS of BBE1 gene was isolated from poppy plant using specific primers. Then, a specific fragment, corresponding to the BBE1 CDS was selected as silencing fragment and cloned into pTZ57R/T plasmid. In the next step, silencing fragment was cloned into pTRV plasmid and resulting constructs (pTRV2-BBE1 and pTRV2-Empty) along with helper plasmid (pTRV1) were transferred to Agrobacterium and infiltrated into young leaves. Result showed a 1608 bp DNA fragment for BBE1 gene. PCR analysis using CP specific primers showed presence of CP transcripts in the infiltrated plants. Real-time PCR analysis showed transcript reduction (about 73%) in the transgenic plant (infiltrated with pTRV2-BBE1) compared with control plants (infiltrated with pTRV2-Empty and non-infiltrated).

Squalen is a polyunsaturated triterpene with wide range of applications in pharmacy. In this study, production of squalen and bioinformatic analysis of corresponding gene were investigated in native strain of Aurantiochytrium. This strain produced 3. 7 and 1. 6 g/l biomass and oil rich in squalene, respectivelly. The amount of produced squalene by this strain was assessed as 48. 9 mg/l. Also, Aurantiochytrium squalen synthase gene was identified and sequenced by molecular analysis. Using the bioinformatics software's, some of codon parameters were evaluated and graphical chart was drawn. By use of modeling databases and their software's, the modeling process of this enzyme was conducted. The results of modeling confirmed the role of some amino acids as alanine, glutamic acid, leucine in construction of alpha helix structure. These types of studies help in identification of squalene synthase reaction mechanism. Evaluation of these hidden information can improve the knowledge of the squeal-like scalene folding process and the challenges of protein expression and will be effective in the new design of the enzyme.

Colon cancer is the fourth most common cancer. With concern to high prevalence and mortality of colon cancer, it is obvious the importance of drug related study, such as effective component in prevention, therapy and recurrence of the cancer. Curcumin is natural yellow pigment isolated from rhizome of turmeric. Curcumin is a new candidate for anticancer treatment, but its low bioavailability and water solubility represent the main disadvantages of its use. In this study, the role of dendrosomal nano carrier assessed in increment of curcumin effects in colon cancer cell line (caco-2). For this reason, IC50 was determined by MTT assay. Also, the effects of dendrosomal nanocurcumin recognized with gene expression study of POU5F1 and NANOG by Real Time PCR. Our finding showed that dendrosomal nano-curcumin decrease cell growth depends on time and concentration, moreover, it significantly declines POU5F1 and NANOG gene expression in colon cancer cell line (caco-2). Overall, the results showed that dendrosomal nano-cucomin could be effective in therapy of colon cancer by reducing the expression of genes involved in unlimited cell proliferation.

Bdellovibrio and like organisms (BALOs) are a group of predatory bacteria of the delta-proteobacteria class, that prey on some gram negative bacteria. In this study, the biological characteristics of a marine BALOs isolate and its application in the elimination of pathogenic bacteria were investigated. A predator bacterium MMHZ1 was isolated from the coastal waters of the Caspian Sea using a double layer agar technique. Proteus that was isolated from the intestines of a fish was used as prey. The isolate was highly motile and bacteriolytic, and had the ablility to form plaque on the double layer agar medium. The 16S rRNA sequencing analysis revealed that the MMHZ1 isolate was similar to Halobacteriovorax litoralis with 99. 62% homology. Biological characterizations revealed that optimum pH, salt concentration and growth temperature of isolate were 7. 2, 0. 24% and 25 ° C, respectively. The results demonstrated that the isolate was grown only in the presence of live prey, however could not utilize the autoclaved dead bacteria as host. In addition, the study of prey range utilization including some gram negative pathogenic bacteria showed that MMHZ1 lysed 71. 42% of the 14 prey tested. The result of this study demonstrated the applicability of using BALOs to biological control of pathogenic bacteria.

Despite significant advances in cancer treatment, interest in the design of new drugs has increased. Some plant peptides show a wide range of cytotoxic activity against cancer cells. The purpose of this study was to investigate the recognition of plant anti-cancer peptides and also to find the most important common features among them. In this regard, a list of the antimicrobial peptides and information about each peptide was extracted. Statistical analyzes were performed using R Studio software. The results showed 55 plant anticancer peptides were taxonomically belonging to the Malpighiales. Approximately length of 44% of peptides was in the range of 25 to 30 amino acids. Histidine and methionine had the lowest abundance among peptide amino acids. Cysteine, serine and glycine were the most abundant amino acids. 91% of peptides had less than 10 acidic amino acids and 71% peptides had less than 10 basic amino acids. A pure charge of 76% peptides was between 2 and-2. 64% of peptides had a Bowman index of less than 1. The low index indicates high hydrophobicity of these peptides and increases their chances for interaction with other proteins. Also, the most important three-dimensional structure of plant anti-cancer peptides was the presence of 3 disulfide bridges. Therefore, pharmaceutical manufacturers and drug designers can use these features extracted from advanced statistical analyzes to synthesize or discover new effective drugs with less side effects.

Pterocarya fraxinifolia is an ancient tree in north of Iran that have been recently reported two small stand of this species in west of Iran, Lorestan and Ilam province. In this study, population diversity and genetic differentiation of west populations and their vulnerability were assessed using ISSR markers. Eight to ten trees were selected from two west population as well as four populations of north of Iran, DNA extracted and genetic diversity was evaluated. AMOVA result indicated that the value of intra and inter population diversity is 88% and 12%, respectively. Shannon index is 0. 157 and mean of heterozygosity is variable 0. 064 to 0. 119. Based on neighbor joining algorithm, the under study populations were split to two main groups that the west populations (Ilam and Lorestan) aa well as Savadkouh were comprised the main group 1. Also, PCoA analysis showed clearly differentiation of Lorestan population from other under study populations.

Thymoquinone (TQ) is an active compound of some medicinal plants. Although different biological and pharmaceutical activities of TQ are well known, its effect on stem cells has not been clarified yet. Therefore, this study was aimed to investigate the effect of TQ on the expression of iNos and Cox-1 genes, which are involved in immunomodulatory potential of mouse mesenchymal stem cells (MSCs), at transcript level. MSCs were isolated from young NMRI mice and their potency was confirmed using the differentiation assay into osteoblasts and adipocytes. The results of MTT assay indicated that the median inhibition concentration (IC50) values of TQ on the MSCs were 8 μ g/ml at 24h and 4 μ g/ml at 48 and 72h after treatment. In addition, more than 90% of TQ-treated MSCs were alive after treatment with concentrations ≤ 2 μ g/ml of TQ for 24h. The results of gene expression analysis by real-time PCR showed that iNos and Cox-1 down-regulated 0. 655 and 0. 615 fold (38. 5 and 34. 5%), respectively, in the TQ-treated MSCs compared to the untreated MSCs (P < 0. 05). In conclusion, this study demonstrates that TQ influences immunomodulatory potential of MSCs. Furthermore, this study provides a good perspective for further efforts in the field and for the use of this compound to affect MSCs capabilities in cell therapy.

Spermatogonial stem cells are the undifferentiated cells in the basal membrane of the seminiferous tubules and are able to produce sperm in male testis. Different experimental studies proved the capatility of undifferentiated spermatogonial stem cells to culture under different conditions, after isolation of the testes. In this study mouse neonate testicular cells (5-7 days-old NMRI strain) isolated by digestive enzymes include Collagenase IV, Dispase and DNAse. Then, approximately 106 cells were cultured in both adherent on MEF feeder layer and non-adherent culture system on 1% agarose with condition mediums that contained growth factors. Studies demonstrates that testicular cells cultured in a non-adherent condition (non-adherent colonies) were able to form colonies similar to spermatogonial stem cell colonies in adherent condition (adherent colonies) and clearly expressed germ cell marker. Electron microscope analyses made evident that both types of colonies were similar to localized spermatogonial stem cells on the basement membrane of seminiferous tubules and showed a high nucleus/cytoplasm ratio. Immunocytochemistry assays proved that both of colony expressed germ cell markers β 1-Integrin, α 6-Integrin and Oct4 positive. Realtime PCR analysis revealed significant differences in the expression of germ cell genes Oct4, Sox-2, CD9 and EPCAM, between these colonies and somatic cells. On the other hand, high expression of Ki67 in these colonies showed that these cells have proliferative characteristics. These results proved that a non-adherent culture system similar to adherent culture could provide a favorable method for in vitro short-term culture of spermatogonial stem-like cell colonies.