Assessment of the ability of Pseudomonas aeruginosa and Staphylococcus aureus to create biofilms during wound healing in a rat model treated with carboxymethyl cellulose/carboxymethyl chitosan hydrogel containing EDTA.

The primary objective of this study was to develop a carboxymethyl cellulose (CMC) and carboxymethyl chitosan (CMCS) hydrogel containing ethylene diamine tetra acetic acid (EDTA) as the materials for wound healing. CMC and CMCS solutions were prepared with a concentration of 4% (w/v). These solutions were made using normal saline serum with a concentration of 0.5% (v/v). Additionally, EDTA with the concentrations of 0.01%, 0.05%, 0.1%, 0.5%, 1%, and 2% (w/v) was included in the prepared polymer solution. The analysis of the hydrogels revealed that they possess porous structures with interconnected pores, with average in size 88.71 ± 5.93 µm. The hydrogels exhibited a swelling capacity of up to 60% of their initial weight within 24 h, as indicated by the weight loss and swelling measurements. The antibacterial experiments showed that the formulated CMC/CMCS/EDTA 0.5% hydrogel inhibited the growth of Staphylococcus aureus and Pseudomonas aeruginosa. Moreover, the produced hydrogels were haemocompatible and biocompatible. At the last stage, the evaluation of wound healing in the animal model demonstrated that the use of the produced hydrogels significantly improved the process of wound healing. Finally, the findings substantiated the effectiveness of the formulated hydrogels as the materials for promoting wound healing and antibacterial agents.

Drug Resistance and Biofilm Formation in Candida Species of Vaginal Origin.

Background: Candida species are normal vaginal flora in healthy women, which can cause vulvovaginal candid-iasis (VVC). The formation of biofilm is a cause of drug resistance in Candida species of vaginal origin. We aimed to specify Candida species cause VVC, detect their biofilm-forming ability, and antifungal susceptibility pattern. Methods: Overall 150 vaginal samples were collected from suspected cases of referring to Bahar Hospital of Shahroud, Iran between Jan 2018 and Jan 2019. Samples were cultured on Sabouraud dextrose agar (SDA), Chrome gar Candida and Corn meal agar (CMA). PCR-RFLP was performed to confirm the identification. Bio-film formation of the identified species was measured by the Crystal Violet method. The susceptibility to fluconazole, clotrimazole, and miconazole was determined based on the CLSI document M27-A3. Results: Of 50 women (33.3%) were suffering from VVC. C.albicans was the predominant species isolated in this study (n=39, 78%) followed by C. glabratia (n=11, 22%). In addition, in 25 (50%) of positive samples, bio-film formation was determined. The mean MIC of fluconazole and clotrimazole for C. albicans was 5.02 µg/mL and 3.92 µg/mL, respectively. Furthermore, the mean MIC related to these drugs for C. glabrata was 12.45 µg / mL and 4.1µg / mL, respectively. The mean diameter of miconazole inhibition zone for C. albicans and C. glabra isolates was 25.13 mm and 24.5mm, respectively and all of them were susceptible to this drug. Conclusion: C.albicans was the predominant Candida species isolated from patients with VVC and also was the predominant biofilm producer species.

Oxidative stress associated with long term occupational exposure to extremely low frequency electric and magnetic fields.

BACKGROUND: Occupational exposure to extremely low frequency electromagnetic fields (ELF-EMFs) may have harmful effects on biologic systems and has raised many concerns in the last decades. OBJECTIVE: The aim of this study was to determine the effects of exposure to extremely low frequency electric and magnetic fields on lipid peroxidation and antioxidant enzyme activities. METHODS: This study was conducted on 115 power plant workers as the exposed group and 145 office workers as the non-exposed group. The levels of Malondialdehyde (MDA), superoxide dismutase (SOD), Catalase (Cat), and total antioxidant capacity (TAC) were measured in the serum of all subjects. Exposure to ELF-EMFs was measured based on spot measurements and the IEEE Std C95.3.1 standard. RESULTS: The levels of MDA, SOD, and Cat in the exposed group were significantly higher than in the non-exposed group. However, the level of TAC was not significantly different between the exposed (2.45±1.02) and non-exposed (2.21±1.07) groups. The levels of MDA and SOD were higher among workers with higher exposure to electric fields than workers with low exposure. All oxidative stress indicators increased with increased exposure to magnetic fields, except TAC. CONCLUSIONS: The antioxidant system imbalance among power plant workers may be related to long term occupational exposure to electromagnetic fields.

The effect of different pulp-capping materials on proliferation, migration and cytokine secretion of human dental pulp stem cells.

OBJECTIVES: Biocompatibility of dental biomaterials plays a critical role in regeneration of dental stem cells. The aim of present study was to evaluate the effects of novel biomaterials of TheraCal-LC (TheraCal; Bisco), Angelus mineral trioxide aggregate (MTA; Angelus), calcium-enriched mixture (CEM; BioniqueDent), and Biodentine (Septodont) on viability of human dental pulp stem cells (hDPSCs). Moreover, the recruitment of dental pulp stem cells is a prerequisite for regeneration of damaged dentin. Therefore, in this study the effects of mentioned biomaterials on migration of hDPSCs and the secretion of some chemoattractive molecules by these cells were examined. MATERIALS AND METHODS: The cell viability of hDPSCs was assessed using MTT assay. Transwell migration assay was used to determine cell migration ability. The cytokine secretion was evaluated using enzyme-linked immunosorbent assay. RESULTS: The biomaterials of MTA, CEM, and Biodentine at different dilutions had no cytotoxic effects on hDPSCs at different time points; however, non-diluted extract of TheraCal showed toxic effects after 24, 48, and 72 hr. Meanwhile, the highest cell migration was observed in the presence of CEM and Biodentine (P<0.05). The secretion of MCP-1 and TGF-ß1 were higher in hDPSCs treated with Biodentine compared to some other groups (P<0.05, P<0.01). Moreover, TheraCal decreased protein secretion of TNF-α (P<0.05), and IL-8 (P<0.01) in hDPSCs. CONCLUSION: The biological compatibility associated with CEM and Biodentine indicates promising applications in the field of vital pulp therapy.

Sulforaphane Modulates Cell Migration and Expression of ß-Catenin and Epithelial Mesenchymal Transition Markers in Breast Cancer Cells.

BACKGROUND: We aimed to assess the effect of sulforaphane (SFN) on breast cancer cell migration and also its effect on the expression of epithelial mesenchymal transition (EMT) markers and ß-catenin. METHODS: This study was performed in Shahroud University of Medical Sciences, Shahroud, Iran from 2017-2018. In this experimental study, MDA-MB-231 cells were treated with different concentrations of SFN (5, 10, 20, 30 and 40 µM) at different time points of 24, 48, and 72 h. The control group was untreated cells. The inhibitory effects of different concentrations of SFN on cell migration at different time points were evaluated using scratch assay. Moreover, apoptosis was assessed by using flow cytometric analysis. The expression of ß-catenin and EMT markers of ZEB1, fibronectin, and claudin-1 were determined by real-time PCR. Western blotting analysis of ß-catenin was applied to determine its changes after SFN treatment. RESULTS: SFN markedly inhibited the migration of cells at concentrations of 10, 20, 30, and 40µM after 24, 48, and 72 h. At relatively, high concentrations (30, 40µM), SFN induced apoptosis. Moreover, SFN reduced the gene expression of ZEB1, fibronectin, and claudin-1 after 72 h. The expression of ß-catenin revealed a time-dependent decrease at the concentration of 40 µM SFN. CONCLUSION: Downregulation of EMT markers and ß-catenin showed accordance with the inhibition of migration. SFN could be a promising drug candidate to reduce metastasis in breast cancer.

Nutrient Deprivation Modulates the Metastatic Potential of Breast Cancer Cells.

BACKGROUND: Breast cancer is the leading cause of cancer related death in women worldwide. The development of metastatic cancer is the main factor contributing to mortality. The molecular mechanisms underlying the metastatic process have yet to be clearly elucidated. However, the interplay between the tumor microenvironment and the cancer cells hold a critical role in influencing the progression of cancer metastasis. Within the microenvironment of solid tumors, the lack of sufficient vasculature leads to the development of nutrient deprived conditions. This study aimed to examine how nutrient deprivation influences factors involved in cancer progression and metastasis. Specifically, we examined how nutrient stress changes cancer cell migration, the gene expression, and cytokine production of metastasis-related factors in a human breast cancer cell line. METHODS: MCF7 breast cancer cells were cultured in serum-free media for 24, 48, and 72 h. Cell migration was evaluated using a transwell migration assay. The transcriptional expression of metastatic related genes was examined via real-time PCR. Cytokine production was examined via enzyme-linked immunosorbent assay. RESULTS: Nutrient deprivation of the MCF7 cells significantly reduced cell migration after 24 h. However, following 72 h of nutrient deprivation, there was significant increase in cell migration compared to the 24 h group. Transcriptional expression of markers involved in migration including, ß-catenin, twist, vimentin, fibronectin, ICAM1, VCAM1, and VEGF were up regulated after 72 h of nutrient deprivation. The cytokines TGFß1, IL-8, and MCP1 were differentially secreted. CONCLUSION: Nutrient deprivation is an environmental stress factor that can influence the behavior of cancer cells. Current treatments implement nutrient deprivation as a potential cancer treatment. Under short periods of nutrient deprivation, cancer cell migration is inhibited. However, our findings show that following extended lengths of nutrient deprivation, cancer cells are capable of adapting themselves to the environmental condition and restoring their migratory abilities. This, in part, may be a result of increased expression of metastasis-related genes. Further research is required to accurately identify how the expression of metastasis-related genes is modulated and controlled in response to nutrient deprivation and environmental stress.

Gene Expression Changes in Pomegranate Peel Extract-Treated Triple-Negative Breast Cancer Cells.

BACKGROUND: Triple-negative breast cancer (TNBC) is treated with highly aggressive non-targeted chemotherapies. Safer and more effective therapeutic approaches than those currently in use are needed. Natural pomegranate peel extract (PPE) has recently been found to inhibit breast cancer progression; however, its mechanisms of action remain unclear. We hypothesized that transcriptional changes in the genes encoding the adherence proteins of intercellular adhesion molecule-1 (ICAM-1) and vascular endothelial growth factor (VEGF), may explain, at least in part, the anti-metastatic properties of PPE. Recently, the tumor microenvironment has been recognized as a key contributor to cancer progression. We speculated that PPE acts by modulating matrix glycoproteins including MMP9 and fibronectin. Moreover, we hypothesized that VEGF, which is required for tumor development, may contribute to the antimetastatic effects of PPE. METHODS: To address these possibilities, MDA-MB-231 cells were treated with different doses of PPE at different time points. Apoptosis was detected by flow cytometry using annexin V and propidium iodide. Cell migration was detected with a transwell assay. Gene expression changes were analyzed by real-time PCR. RESULTS: Exposure to PPE resulted in TNBC cell death and markedly inhibited PPE-resistant cell migration. Moreover, PPE up-regulated the expression of ICAM-1, a protein essential for cell adhesion, and down-regulated the expression of MMP9, fibronectin, and VEGF, the products of which contribute to cancer cell migration. CONCLUSION: Transcriptional changes in ICAM-1, MMP9, fibronectin, and VEGF may contribute to PPE-mediated antimetastatic effects in TNBC.

Pomegranate peel extract inhibits expression of ß-catenin, epithelial mesenchymal transition, and metastasis in triple negative breast cancer cells.

The standard treatment for triple-negative breast cancer (TNBC) is chemotherapy, which is highly toxic to patients; thereby, there is a need to identify safer and more effective therapeutic approaches. Medicinal plants constitute a common alternative for cancer treatment. Pomegranate is a well-known fruit in this context, but its antimetastatic property has not been extensively studied. As breast cancer-related deaths from TNBC are mainly due to metastasis, the present study was designed to investigate the antimigratory effect of pomegranate peel extract (PPE) on TNBC cells. For this purpose, the MDA-MB-231 cells were treated with different concentrations of PPE for 24, 48 and 72 hr. The effects of PPE on cell migration and invasion were determined by wound healing and transwell assays. To address the possible molecular mechanisms underlying the antimetastatic effect of PPE, real-time quantitative PCR analysis of selected epithelial mesenchymal transition (EMT) markers were performed. Moreover, the expression of ß-catenin as a critical factor in promoting cancer metastasis was examined. PPE markedly inhibited the migration and invasion of cells at concentrations of 25, 50, 100, 250, 500, and 1000µg/ml. At relatively high concentrations (500, 1000µg/ml), PPE induced apoptosis. Moreover, PPE decreased the gene expression of vimentin, ZEB1, and ß-catenin and also increased the expression of E-cadherin in TNBC cells. The protein level of ß-catenin, as measured using western analysis, revealed a time-dependent decrease at the concentration of 1000µg/ml PPE. Downregulation of EMT markers and ß-catenin showed accordance with the inhibition of migration and invasion. The present data show that PPE could be a promising drug candidate to reduce metastasis in TNBC cells.

Effects of Two Calcium Silicate Cements on Transforming Growth Factor-ß1 Secretion from Human Dental Pulp Stem Cells.

Introduction: The aims of this in vitro study were to evaluate the effects of two calcium silicate based cements, Calcum-enriched Mixture (CEM) and Biodentine on proliferation of human dental pulp stem cells (hDPSCs) and the effects of proposed cements on the secretion of Transforming Growth Factor ß1 (TGF-ß1). Methods and materials: The cell cultures of human Dental Pulp Stem Cells (hDPSCs) at passage 3-5 were treated with various dilutions (1/1, 1/2, 1/4, 1/8, 1/16, and 1/32) of CEM and Biodentine extracts to assess the cell proliferation using 3-(4, 5-dimethylthiazol-2-Y1)-2, 5-diphenyltetrazolium brovide (MTT) assay after 48 and 72 h. The amount of TGF-ß 1 secretion were estimated after 72 h using an enzyme-linked immunosorbent assay. Data were analyzed using the one-way analysis of variance (ANOVA) followed by the Dunnett's test at the level of significance set at 0.05. Result: CEM showed the highest rates of cell proliferation compared to Biodentine after 72 h (P<0.05). A greater amount of TGF-ß1 was secreted by hDPSCs treated with Biodentine compared to CEM (P<0.05). These differences were statistically significant (P<0.05). Conclusion: In this in vitro study hDPSCs showed more proliferation capacity with CEM rather than Biodentine and TGF-ß1 secretion rate in Biodentine was higher.

Appropriate anthropometric indices of obesity and overweight for diagnosis of metabolic syndrome and its relationship with oxidative stress.

AIMS: this study aimed to determine the most appropriate anthropometric indices for diagnosis of metabolic syndrome (MetS) and its relationship with oxidative stress markers. MATERIAL AND METHODS: This cross-sectional study was conducted in 2015 on 108 employees working in Shahroud University of Medical Sciences. Demographics, anthropometric indices (BMI: Body mass index; WC: Waist circumference, WHR: Waist hip ratio, WHtR: Waist-to-height Ratio), Mets: and Then oxidative stress markers (total antioxidant capacity; TAC, Malondialdehyde; MDA, serum superoxide dismutase; SOD, catalase; Cat) were measured. All analyses were performed at a significant level of 0.05, using the SPSS Statistics 21 and Stata 12 software. RESULT: The mean age of the participants was 41.4±7.8years. the mean values of different anthropometric indices in patients with metabolic syndrome were higher than those in subjects without MetS and this difference was significant. According to ROC curve the best marker for diagnosis of Mets was WHtR (Waist-to-height Ratio) and its cut off point was 0.54.Also, there was a positive correlation between WHtR and MDA serum levels. In addition, there was a negative correlation between WHtR and the levels of TAC, SOD, and CAT but it was not significant. CONCLUSIONS: It seems that WHtR can be a valuable marker for predicting metabolic disorders and related diseases; moreover, it can be used for evaluation of oxidative stress level. Finally, the formula WC=height×0.54 as a simple tool for prevention of metabolic diseases can be used in university personnel.

The Effects of Berberine and Palmatine on Efflux Pumps Inhibition with Different Gene Patterns in Pseudomonas aeruginosa Isolated from Burn Infections.

BACKGROUND: Related Multidrug Resistance (MDR) to efflux pumps is a significant problem in treating infections caused by Pseudomonas aeruginosa (P. aeruginosa). Plant compounds have been identified as Pump Inhibitors (EPIs). In the current study, the potential effect of Berberine and Palmatine as EPIs were investigated on efflux pump inhibition through focusing on different gene patterns in P. aeruginosa isolated from burn infections. METHODS: All isolates were collected and identified using the standard biochemical tests. Antimicrobial sensitivity was performed based on disk agar diffusion method for 12 antibiotics. MIC-MBC tests were also performed based on the broth microdilution method to detect synergistic relationship between ciprofloxacin, Berberine and Palmatine. Detection of mexA, mexB, mexC, mexD, mexE, mexF and mexX was conducted by PCR assay. Fisher's Exact test and Logistic Regression were used as statistical tools. RESULTS: A total of 60 P. aeruginosa isolates were collected. The highest and lowest levels of resistance were found to be respectively against clindamycin and tigecycline. Comparing the MIC with MBC distribution, it was found that Berberine and Palmatine lower the MIC-MBC level of ciprofloxacin. The PCR results indicated that the highest frequency is about MexAB-OprM operon. The statistical analysis among different gene patterns of efflux pumps showed that there were no significant relationships between the effectiveness of Berberine and Palmatine (p>0.05). CONCLUSION: It can be speculated that Berberine and Palmatine both act as EPIs and can be used as auxiliary treatments with the purpose of increasing the effect of available antibiotics as well as decreasing the emergence of MDR bacteria. The efficiency of these combinations should be studied further under in vivo conditions to have a more comprehensive conclusion regarding this issue.

Association between serum uric acid (SUA) levels and metabolic syndrome (MetS) components in personnel of Shahroud University of Medical Sciences.

AIMS: Serum uric acid level has been suggested to be associated with metabolic syndrome risk factors. However, the association between metabolic syndrome and serum uric acid is still controversial and challenging. This study was aimed to investigate the association between serum uric acid levels and metabolic syndrome components in personnel of the Shahroud University of Medical Sciences. MATERIAL AND METHODS: This case-control study was conducted on 499 personnel aged 30-60 years old who were working in Shahroud University of Medical Sciences, in 2015. MetS was defined according to the National Cholesterol Education Program (NCEP) criteria. The relationship between serum UA level and the number of metabolic components was determined by linear regression analysis. RESULT: In this study, the mean concentration of serum uric acid in men with the syndrome was higher than that in women. Mean serum UA level increased as the number of metabolic factors increased. The mean serum uric acid levels was 4.98±1.64 in patients with metabolic syndrome and 4.5±1.28 in non-patients (p=0.005). Subject with abnormal uric acid were almost 2.62 times more likely than other subject to develop the syndrome. CONCLUSIONS: The results of this study showed that only hypertriglyceridemia is a component which increases the risk of hyperuricemia. In addition, hyperuricemia increases the risk of metabolic syndrome by more than two fold. It seems that high uric acid can be considered as a predisposing factor for metabolic syndrome; thus, it is recommended to measure serum uric acid in routine tests.