The Effect of Placenta Growth Factor Knockdown on hsa-miR-22-3p, hsa-let-7b-3p, hsa-miR-451b, and hsa-mir-4290 Expressions in MKN-45- derived Gastric Cancer Stem-like Cells

Background: Placental growth factor is involved in human gastric cancer initiation and progression through stimulating the proliferation, angiogenesis, invasion and metastasis of cancerous cells. Previous studies indicate that the expression profiles of hsa-miR-22-3p, hsa-let-7b-3p, hsa-miR-451b, and hsa-mir-4290 change in MKN-45- derived gastric cancer stem-like cells. Therefore, this study aims to investigate the effect of PlGF knockdown on hsa-miR-22-3p, hsa-let-7b-3p, hsa-miR-451b, and hsa-mir-4290 expressions in MKN-45-derived gastric cancer stem-like cells. Methods: We used a non-adhesive culture system to derive the cancer stem-like cells from MKN-45 cells. PlGF gene silencing was performed by PlGF-specific siRNA. The transcript of PlGF and miRNAs were measured by real-time RT-PCR. We conducted bioinformatics analyses with the online software programs TargetScan, miRanda, miRWalk, PicTar, and the Database for Annotation, Visualization, and Integrated Discovery tools to predict miRNAs’ targets and their signaling pathways. Results: hsa-let-7b-3p had a 2.28-fold up-regulation, whereas we observed downregulation of hsa-mir-451b (25%), hsa-mir-4290 (34%), and hsa-mir-22-3p (9%). Bioinformatics analysis results indicated that the miRNA target genes TGF-β, MAPK, and Wnt, and hedgehog signaling pathways contributed to cancer initiation and progression by influencing different cellular behaviors. Conclusion: We suggest that PlGF signaling may influence miRNA expression profiles in MKN-45-derived cancer stem-like cells, which can influence the expressions of different genes and signaling pathways. However, more empirical studies should determine the exact effect of PlGF knockdown on the expression of miRNA targets in cancer stem-like cells to locate their actual gene targets

The effect of resveratrol on oxidative stress in the liver and serum of a rat model of polycystic ovary syndrome: An experimental study

Background: Studies of oxidative status in polycystic ovarian syndrome (PCOS) patients are limited with inconsistent results. The effects of resveratrol as a natural antioxidant on oxidative status in PCOS aren’t clear. Objective: This study evaluated effects of resveratrol on oxidative stress in the liver and serum of the PCOS rats. Materials and Methods: Fifteen female Wistar rats (3 wk old) were divided into 3 groups (n=5/each e): Control group, PCO-Control group, and PCO-Resveratrol group. For induction of polycystic ovary phenotype, testosterone enanthate 10 mg/kg was injected for 35 days subcutaneously. Then, resveratrol 10 mg/kg was injected intraperitoneally for 28 days to rats of the PCO-Resveratrol group. Ovarian sections were stained with hematoxylin/eosin. The serum glucose and insulin and the levels of malondialdehyde (MDA) and total antioxidant capacity (TAC) in serum and liver were measured. Results: Control animals showed normal ovarian morphology and PCO-Control animals exhibited cystic follicles. There were no significant differences in liver TAC between groups. The serum MDA (p=0.034), and homeostatic model assessment insulin resistance (HOMA-IR) (p=0.014) levels in PCO-Control rats were higher than the controls. The liver MDA in PCO-Control rats was more than that of controls (p=0.001). The HOMA-IR (p=0.008) and serum MDA (p=0.006) levels in PCO-Control rats were more than those of PCO-Resveratrol rats (p=0.008). In PCO-Resveratrol group, serum TAC was higher than that of PCO-Control group (p=0.022) and liver MDA was more than controls (p=0.01). Conclusion: Results indicated that the induction of PCOS in rats increased lipid peroxidation and insulin resistance and resveratrol improved these complications

Biomedical applications of MnO2 nanomaterials as nanozyme-based theranostics

Manganese dioxide (MnO2) nanoenzymes/nanozymes (MnO2-NEs) are 1–100 nm nanomaterials that mimic catalytic, oxidative, peroxidase, and superoxide dismutase activities. The oxidative-like activity of MnO2-NEs makes them suitable for developing effective and low-cost colorimetric detection assays of biomolecules. Interestingly, MnO2-NEs also demonstrate scavenging properties against reactive oxygen species (ROS) in various pathological conditions. In addition, due to the decomposition of MnO2-NEs in the tumor microenvironment (TME) and the production of Mn2+, they can act as a contrast agent for improving clinical imaging diagnostics. MnO2-NEs also can use as an in situ oxygen production system in TME, thereby overcoming hypoxic conditions and their consequences in the progression of cancer. Furthermore, MnO2-NEs as a shell and coating make the nanosystems smart and, therefore, in combination with other nanomaterials, the MnO2-NEs can be used as an intelligent nanocarrier for delivering drugs, photosensitizers, and sonosensitizers in vivo. Moreover, these capabilities make MnO2-NEs a promising candidate for the detection and treatment of different human diseases such as cancer, metabolic, infectious, and inflammatory pathological conditions. MnO2-NEs also have ROS-scavenging and anti-bacterial properties against Gram-positive and Gram-negative bacterial strains, which make them suitable for wound healing applications. Given the importance of nanomaterials and their potential applications in biomedicine, this review aimed to discuss the biochemical properties and the theranostic roles of MnO2-NEs and recent advances in their use in colorimetric detection assays of biomolecules, diagnostic imaging, drug delivery, and combinatorial therapy applications. Finally, the challenges of MnO2-NEs applications in biomedicine will be discussed

Biological removal of nickel (II) by Bacillus sp. KL1 in different conditions: optimization by Taguchi statistical approach

Bioremediation is the removal of heavy-metals such as nickel (Ni) using microorganisms and has been considered as an important field in the biotechnology. Isolation and characterization of microorganisms exhibiting bioremediation activities and their optimization to treat polluted wastewaters is a vital and difficult task in remediation technologies. In this study, investigation was carried out to isolate Ni (II) remediating microbial strains from soils contaminated with municipal solid waste leachate. Furthermore, Taguchi design of experiments were used to evaluate the influence of concentration, pH, temperature, and time on bioremediation of Ni (II) using isolated bacteria. This study concluded that Bacillus sp. KL1 is a Ni (II)-resistant strain and had Ni (II) bioremediation activity. The highest bioremediation of Ni (II) was observed as 55.06% after 24 h at 30ºC, pH 7, and 100 ppm concentration. Moreover, it was also observed that concentration is the most effective factor in the bioremediation process. In conclusion, we have demonstrated that bacteria isolated from soils contaminated with garbage leachate have the Bacillus sp. KL1 bacteria which can efficiently uptake and eliminate Ni (II) from contaminated sites and thus makes it possible to treat heavy-metal containing wastewaters in industry by using this microorganism at optimized conditions

Evaluation of Tnf-α and Il-6 mRNAs expressions in visceral and subcutaneous adipose tissues of polycystic ovarian rats and effects of resveratrol

Objective(s): Some studies suggest that chronic low-grade inflammation is involved in insulin resistance in polycystic ovary syndrome (PCOS). This study assessed possible involvement of alteration in expression of two pro-inflammatory factors, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in adipose tissues of PCOS rats in the impairment of insulin actions. Also, effects of resveratrol as an anti-inflammatory agent were investigated. Materials and Methods: Fifteen female Wistar rats (21 days old) were divided into three groups (n=5): Ι) Control, П) PCO-model-saline: served as PCOS rats and to induce PCOS, received subcutaneously testosterone enanthate 1 mg/100 g body weight subcutaneously for 35 days, Ш) PCO-model-resveratrol, after receiving testosterone, received resveratrol 10 mg/kg intraperitoneally for 28 days. The expression of Tnf-α and Il-6 mRNAs in adipose tissues was determined by the qRT-PCR method. Results: The Il-6 mRNA expression in the visceral adipose tissue of PCOS rats was increased in comparison to controls (

Amino acid profile changes during enrichment of spheroid cells with cancer stem cell properties in MCF‐7 and MDA‐MB‐231 cell lines

Abstract Background Cancer stem cells (CSCs), subpopulations of cancer cells, are responsible for tumor progression, metastasis, and relapse. Changes in amino acid metabolism are linked to breast cancer recurrence and metastasis. Aims This study aimed to evaluate the changes in the amino acid profile in MCF‐7 and MDA‐MB‐231 cells during spheroid formation to discover the specific metabolic properties in CSCs. Methods MCF‐7 and MDA‐MB‐231 breast cancer cells were cultured as spheroids and evaluated to characterize their CSC properties. The characteristics of CSC were evaluated by examining the expression of CSC markers and conducting drug resistance assays. In addition, amino acid profile change during the enrichment of breast cancer stem cells in the spheroids was investigated by high‐performance liquid chromatography (HPLC). Results The results indicated that out of 20 different amino acids analyzed, 19 of them decreased during the spheroid formation process. Alanine, lysine, phenylalanine, threonine, and glycine showed significant reductions in the conditioned media of both cell lines in the spheroid form compared to the monolayer cells. Only one of the amino acids increased in MCF‐7 and MDA‐MB‐231 spheroids (histidine and serine, respectively). Conclusion Our results suggest that certain amino acids identified in this study can be used for a better understanding of the molecular mechanisms associated with breast cancer stem cell formation

Establishment of a culture condition for strong proliferation and enrichment of chicken spermatogonial stem cells in vitro

Poultry spermatogonial stem cells (SSCs) have the potential to serve as a model for studying the basic biology of SSC and they can also be used for biotechnological purposes. However, the small number of SSCs and the presence of the testicular somatic cells with SSCs have limited their applications. Therefore, this study was undertaken for the first time to investigate the effect of a serum-free medium supplemented with a combination of specific growth factors and B27 on the proliferation and enrichment of newborn chicken SSCs in vitro. Newborn chicken testicular cells were cultured in a serum-free DMEM, supplemented with GDNF, bFGF, LIF, and EGF growth factors and also B27 as an alternative for FBS. Presence and maintenance of the SSCs in the enriched cultures were evaluated by detection of alkaline phosphatase (AP) activity and ASZ1, POU5F1, CVH and GPR125 gene expression. A small number of clusters and colonies were emerged in testicular cell cultures before treatment with the enriched cell culture medium. Enrichment of the DMEM with the above indicated factors strongly promoted the proliferation of the chicken SSCs. Moreover, this culture condition declined attachment and maintenance of the testicular somatic cells and thus they decreased gradually in the cultures. The enriched SSCs were positive for AP activity and with detectable levels of ASZ1, POU5F1, CVH and GPR125 gene expression. This study shows that serum-free medium supplemented with a combination of B27 and the above indicated growth factors induces proliferation and enrichment of chicken SSCs in vitro in a short period of time

Construction and Quantitative Evaluation of a Dual Specific Promoter System for Monitoring the Expression Status of Stra8 and c-kit Genes.

Applications of genetic constructs with multiple promoters, which are fused with reporter genes and simultaneous monitoring of various events in cells, have gained special attention in recent years. Lentiviral vectors, with their distinctive characteristics, have been considered to monitor the developmental changes of cells in vitro. In this study, we constructed a novel lentiviral vector (FUM-M), containing two germ cell-specific promoters (Stra8 and c-kit), fused with ZsGreen and DsRed2 reporter genes, and evaluated its efficiency in different cells following treatments with retinoic acid and DMSO. Several cell lines (P19, GC-1 spg and HEK293T) were transduced with this vector, and functional capabilities of the promoters were verified by flow cytometry and quantitative RT-PCR. Our results indicate that FUM-M shows dynamic behavior in the presence and absence of extrinsic factors. A correlation was also observed between the function of promoters, present in the lentiviral construct and the endogenous level of the Stra8 and c-kit mRNAs in the cells. In conclusion, we recommend this strategy, which needs further optimization of the constructs, as a beneficial and practical way to screen chemical inducers involved in cellular differentiation toward germ-like cells