Screening and identification of DNA nucleic acid aptamers against F1 protein of Yersinia pestis using SELEX method.

BACKGROUND: Yersinia pestis is a bacterium that causes the disease plague. It has caused the deaths of many people throughout history. The bacterium possesses several virulence factors (pPla, pFra, and PYV). PFra plasmid encodes fraction 1 (F1) capsular antigen. F1 protein protects the bacterium against host immune cells through phagocytosis process. This protein is specific for Y. pestis. Many diagnostic techniques are based on molecular and serological detection and quantification of F1 protein in different food and clinical samples. Aptamers are small nucleic acid sequences that can act as specific ligands for many targets.This study, aimed to isolate the high-affinity ssDNA aptamers against F1 protein. METHODS AND RESULTS: In this study, SELEX was used as the main strategy in screening aptamers. Moreover, enzyme-linked aptamer sorbent assay (ELASA) and surface plasmon resonance (SPR) were used to determine the affinity and specificity of obtained aptamers to F1 protein. The analysis showed that among the obtained aptamers, the three aptamers of Yer 21, Yer 24, and Yer 25 were selected with a KD value of 1.344E - 7, 2.004E - 8, and 1.68E - 8 M, respectively. The limit of detection (LoD) was found to be 0.05, 0.076, and 0.033 µg/ml for Yer 21, Yer 24, and Yer 25, respectively. CONCLUSION: This study demonstrated that the synthesized aptamers could serve as effective tools for detecting and analyzing the F1 protein, indicating their potential value in future diagnostic applications.

Evaluation of anti-biofilm activity of Lactobacillus rhamnosus GG and Nisin on the expression of aap, ica-A and ica-D as biofilm-associated genes of Staphylococcus epidermidis.

Background and Objectives: In the present study, the anti-biofilm activity of Lactobacillus rhamnosus GG and Nisin was investigated on biofilm-forming abilities of Staphylococcus epidermidis strains and the expression of the biofilm-associated genes. Materials and Methods: In this study, the standard strain of L. rhamnosus GG (ATCC 53103) and Nisin were used to assess their anti-microbial and anti-biofilm effects on S. epidermidis (RP62A). Results: The MIC and MBC analysis showed that Nisin at 256 µg/mL and 512 µg/mL, and L. rhamnosus GG at 1×107 CFU/mL and 1×108 CFU/mL have anti-microbial activity compared to the negative control respectively. L. rhamnosus GG bacteria and Nisin inhibited the biofilm formation of S. epidermidis based on optical density of at 570 nm (P <0.001). The relative mRNA expression of aap, icaA, and icaD genes was significantly reduced compared to the negative control after treating S. epidermidis with sub-MIC of Nisin (0.44, 0.25 and 0.6 fold, respectively) (P>0.05). In addition, the relative expression of aap and icaA genes, but not icaD (P>0.05), was significantly lower than the negative control (0.62 and 0.7 fold, respectively) (P>0.05), after exposure to the sub MIC of L. rhamnosus GG. Conclusion: Nisin and L. rhamnosus GG exhibit potent activity against biofilm-forming abilities of S. epidermidis and these agents could be utilized as an anti-biofilm agents against S. epidermidis infections.

The Anti-Adhesion Effect of Nisin as a Robust Lantibiotic on the Colorectal Cancer Cells.

Background: Bacteriocins are a type of antimicrobial peptide that are produced by probiotics. They have been studied as possible therapeutic drugs and have been used to suppress bacterial development in foods. Nisin is a potent bacteriocin having the anti-microbial and anti-cancer characteristics produced by Lactococcus lactis. The aim of the present paper is to evaluate the influence of Nisin on cell adhesion and its two related genes, mmp-2 and mmp-9, in the colorectal cancer cell line. Materials and Methods: For this purpose, HT-29 cells were treated with various concentrations of Nisin and the cell cytotoxicity, cell adhesion, and gene expression were evaluated using the MTT assay, cell adhesion assay, and real-time PCR. Results: Our findings showed that 32 to 1024 µg/ml of Nisin resulted in a significant reduction in cell viability (P < 0.05). Furthermore, 128 and 256 µg/ml of Nisin significantly reduced the cell adhesion, and mmp-2 and mmp-9 gene expressions (P < 0.05). Conclusion: Our findings suggested that Nisin could prevent metastasis and cancer progression.

Exercise Training and Probiotic Lacticaseibacillus rhamnosus GG Reduce Tetracycline-Induced Liver Oxidative Stress and Inflammation in Rats with Hepatic Steatosis.

Lifestyle modification with regular exercise can improve metabolic diseases by reducing lipid profile and inflammation. Probiotics have been recently recommended not only for gastrointestinal diseases but also for metabolic and even degenerative diseases. Therefore, in the present study, the effect of high-intensity interval training (HIIT) and Lacticaseibacillus rhamnosus strain GG (LGG) as a probiotic on tetracycline-induced hepatic steatosis in an animal model was evaluated. Eighty male Wistar rats were randomly divided into eight groups (n = 10 in each group): control, LGG, HIIT, LGG + HIIT, tetracycline-induced (TTC), TTC + LGG, TTC + HIIT, and TTC + LGG + HIIT. The rats are treated by intraperitoneal injection (IP) with 140 mg kg-1 tetracycline, an antibiotic previously known to induce steatosis. The exercise training groups performed HIIT 5 days/week for 5 weeks, and 107 CFU/ml of Lacticaseibacillus rhamnosus GG was gavaged for the LGG groups 5 days/week for 5 weeks. Fatty droplets in the hepatocyte were considered with Oil Red staining. TTC-receiving rats have more lipid accumulation and larger lipid droplets in the liver compared to healthy animals. The two-way ANOVA showed that the interaction of LGG and HIIT significantly decreased LDL, cholesterol, and triglyceride in the healthy rats (p < 0.05). In TTC-receiving rats, the interaction of LGG and HIIT significantly increased HDL and SOD and significantly decreased triglyceride, ALP, AST, and ALT (p < 0.05). The consumption of probiotic LGG, along with HIIT with control of lipid profile and liver enzymes and improvement of the oxidative capacity, neutralizes the damage of TTC to liver tissue. Therefore, this protocol can be recommended for people with hepatic steatosis.

Screening and Identification of DNA Nanostructure Aptamer Using the SELEX Method for Detection of Epsilon Toxin.

Background: Epsilon toxin (ETX), produced by Clostridium perfringens, is one of the most potent toxins known, with a lethal potency approaching that of botulinum neurotoxins. Epsilon toxin is responsible for enteritis. Therefore, the development of rapid and simple methods to detect ETX is imperative. Aptamers are single-stranded oligonucleotides that can bind tightly to specific target molecules with an affinity comparable to that of monoclonal antibodies (mAbs). DNA aptamers can serve as tools for the molecular identification of organisms, such as pathogen subspecies. Objectives: This study aimed to isolate high-affinity single-stranded DNA (ssDNA) aptamers against ETX. Methods: This study identified aptamers using the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method, enzyme-linked apta-sorbent assay (ELASA), and surface plasmon resonance (SPR) to determine the affinity and specificity of the newly obtained aptamers targeting ETX. Results: Several aptamers obtained through the SELEX process were studied. Among them, 2 aptamers, ETX clone 3 (ETX3; dissociation constant (Kd = 8.4 ± 2.4E-9M) and ETX11 (Kd = 6.3 ± 1.3E-9M) had favorable specificity for ETX. The limits of detection were 0.21 and 0.08 µg/mL for ETX3 and ETX11, respectively. . Conclusions: The discovered aptamers can be used in various aptamer-based rapid diagnostic tests for the detection of ETX.

Effect of extracellular vesicles of Lactobacillus rhamnosus GG on the expression of CEA gene and protein released by colorectal cancer cells.

Background and Objectives: Almost all living cells secret nano-sized structures enclosed by the lipid bilayer called extracellular vesicles (EVs) into their extracellular milieu. These EVs play important roles in several physiological processes as a cargo delivery system. In probiotics, EVs are the main communication tool with the host. The present study aimed to assess the effect of EVs originated from Lactobacillus rhamnosus GG on the Carcinoembryonic antigen (cea) gene expression and protein (CEA) synthesis in the SW480 and HT-29 cell lines. Materials and Methods: Different concentrations of Lactobacillus rhamnosus GG EVs were applied on the SW480 and HT-29 cell lines. The MTT assay, Real-Time PCR, and ELISA analysis methods were exploited to explore the cell viability and the expression level of the cea gene in comparison with the ß-actin gene as the control. Results: The two concentrations of 80 and 100 µg/ml of Lactobacillus rhamnosus GG EVs considerably affected the anti-proliferation and increased the amount of both CEA mRNA and protein (p < 0.05). Conclusion: Our findings showed that EVs of Lactobacillus rhamnosus GG could induce the gene expression and protein synthesis of CEA. Also, they reduced the cell proliferation of HT29 and SW480. Thus, probiotics such as EVs of Lactobacillus rhamnosus GG could be useful for preventing colorectal cancer.

Targeting the EGFR in cancer cells by fusion protein consisting of arazyme and third loop of TGF-alpha: an in silico study.

The anticancer effects of arazyme, a bacterial metalloprotease, have been revealed in previous studies. Because of the overexpression of epidermal growth factor receptor (EGFR) in tumor cells, targeting this receptor is one of the approaches to cancer therapy. In the present study, we designed fusion protein by using a non-mitogenic binding sequence of TGFα, arazyme, and a suitable linker. The I-TASSER and Robetta web servers were employed to predict the territory structures of the Arazyme-linker-TGFαL3, and TGFαL3-linker-Arazyme. Then, models were validated by using PROCHECK, ERRAT, ProSA, and MolProbity web servers. After docking to EGFR, Arazyme-linker-TGFαL3 showed a higher binding affinity and was selected to be optimized through 100 ns Molecular dynamic (MD) simulation. Next, the stability of ligand-bound receptor was examined utilizing MD simulation for 100 ns. Furthermore, the binding free energy calculation and free energy decomposition were carried out employing MM-PBSA and MM-GBSA methods, respectively. The root mean square deviation and fluctuation (RMSD, RMSF), the radius of gyration, H-bond, and binding free energy analysis revealed the stability of the complex during simulation time. Finally, linear and conformational epitopes of B cells, as well as MHC class I and MHC class II were predicted by using different web servers. Meanwhile, the potential B cell and T cell epitopes were identified throughout arazyme protein sequence. Collectively, this study suggests a novel chimera protein candidate prevent cancer cells potentially by inducing an immune response and inhibiting cell proliferation. Communicated by Ramaswamy H. Sarma.

ARA-linker-TGFαL3: a novel chimera protein to target breast cancer cells.

Targeted cancer therapies based on overexpressed receptors and the fractions containing immunotoxins and bacterial metabolites are one of the well-known methods to overcome the chemotherapy resistance of cancer cells. In this paper, we designed ARA-linker-TGFαL3, using Arazyme, a Serratia proteamaculans metabolite, and a third loop segment of TGFα to target EGFR-expressing breast cancer cells. After cloning in pET28a (+), the expression of recombinant protein was optimized in Escherichia coli strain BL21 (DE3). MDA-MB-468 (EGFR positive) and MDA-MB-453 (EGFR negative) breast cancer cell lines were employed. Also, the chemotherapeutic drug, Taxotere (Docetaxel), was employed to compare cytotoxicity effects. Cell ELISA assessed the binding affinity of recombinant proteins to the receptor, and the cytotoxicity was detected by MTT and lactate dehydrogenase release assays. The interfacing with cancer cell adhesion was evaluated. Furthermore, the induction of apoptosis was examined utilizing flow cytometric analysis, and caspase-3 activity assay. Moreover, RT-PCR was conducted to study the expression of apoptosis (bax, bcl2, and casp3), angiogenesis (vegfr2), and metastasis (mmp2 and mmp9) genes. ARA-linker-TGFαL3 revealed a higher binding affinity, cytotoxicity, and early apoptosis induction in MDA-MB-468 cells compared to the effects of Arazyme while both recombinant proteins showed similar effects on MDA-MB-453. In addition, the Taxotere caused the highest cytotoxicity on cancer cells through induction of late apoptosis. Meanwhile, the expression of angiogenesis and metastasis genes was decreased in both cell lines after treatment with either ARA-linker-TGFαL3 or Arazyme. Our in vitro results indicated the therapeutic effect of ARA-linker-TGFαL3 on breast cancer cells.

Positive Association of Matrix Proteins Alteration with TAZ and The Progression of High-Grade Bladder Cancer.

OBJECTIVE: Bladder cancer is the 9th leading cause of human urologic malignancy and the 13th cause of death worldwide. Increased collagen cross-linking, NIDOGEN1 expression and consequently stiffness of extracellular matrix (ECM) may be responsible for the mechanotransduction and regulation of transcriptional co-activator with PDZ-binding motif (TAZ) and transforming growth factor ß1 (TGF-ß1) signaling pathways, resulting in progression of tumorigenesis. The present study aimed to assess whether type 1 collagen expression is associated with TAZ nuclear localization. MATERIALS AND METHODS: In this case-control study, real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemical analysis were performed to evaluate the activation of the TAZ pathway in patients with bladder cancer (n=40) and healthy individuals (n=20). The ELISA method was also conducted to measure the serum concentrations of TGF-ß1. Masson's trichrome staining was carried out to histologically evaluate the density of type 1 collagen. RESULTS: Our findings that the expression levels of COL1A1, COL1A2, NIDOGEN1, TAZ, and TGF-ß1 genes were overexpressed in patients with bladder cancer, and their expression levels were positively associated with the grade of bladder cancer. The immunohistochemical analysis demonstrated that the nuclear localization of TAZ was markedly correlated with high-grade bladder cancer. We also found that TAZ nuclear localization was substantially higher in cancerous tissues as compared with normal bladder tissues. Masson's trichrome staining showed that the tissue density of type I collagen was considerably increased in patients with bladder cancer as compared with healthy subjects. CONCLUSION: According to our findings, it seems the alterations in the expression of type I collagen and NIDOGEN1, as well as TAZ nuclear localization influence the progression of bladder cancer. The significance of TGF-ß1 and TAZ expression in tumorigenesis and progression to high-grade bladder cancer was also highlighted. However, a possible relationship between TGF-ß1 expression and the Hippo pathway needs further investigations.

The effect of Propolis on inhibition of Aspergillus parasiticus growth, aflatoxin production and expression of aflatoxin biosynthesis pathway genes.

BACKGROUND AND PURPOSE: Aflatoxins are one of the most important mycotoxins, which have been classified as Group I carcinogenic compounds by the International Agency for Research on Cancer. This investigation aimed to examine the effect of Propolis on inhibition of the Aspergillus parasiticus growth, aflatoxin production and expression of aflatoxin biosynthesis pathway genes. MATERIALS AND METHODS: A standard strain of Aspergillus parasiticus (ATCC 15517) was used to perform antifungal susceptibility test, using a microdilution method in accordance with the CLSI M38-A2 guidelines. The aflatoxin concentrations in the control and treated media were determined by HPLC. Also, the quantitative changes in the level of nor-1, ver-1 and omtA genes expression in aflatoxin biosynthetic pathway were analyzed using Real-Time PCR method. RESULTS: The results showed that the minimum inhibitory concentrations (MIC) of propolis was 100 µg/ml. The results showed that total levels of aflatoxin decreased from 386.1 ppm to 3.01 ppm at 50 µg/ml of propolis. In addition, quantitative real-time PCR analysis showed that the level of nor-1, ver-1 and omtA genes expression was significantly decreased after treatment with propolis extract. CONCLUSIONS: The findings reveal that propolis extract, have a significant inhibitory effect on important genes for aflatoxin biosynthesis pathway in aflatoxin production.

Breast cancer targeted/ therapeutic with double and triple fusion Immunotoxins.

Target-specific transport of therapeutic agents holds promise to increase the efficacy of cancer treatment by decreasing injury to normal tissues and post treatment problems. HER2 is a tumor cell surface marker that is expressed in 25-30 % of breast cancer patients. The significant role of HER2 in cancer development and its biological feature makes it a highly appealing goal for the therapeutic treatment of cancer targeted therapy using HER2 monoclonal antibody. This approach is currently used as a special treatment against breast cancer in some research. In the present study, HER2 monoclonal antibody (mAb), (Herceptin) fused to PE38 by recombinant DNA technology and a new recombinant IT was developed. The scFv(Herceptin)-PE-STXA and scFv(Herceptin)-PE fusions cloned in pET28a and recombinant protein expression was carried out and then the proteins were purified. MCF-7 and SKBR-3 cells were used as HER2-negative and HER2-positive breast cancer cells, respectively. The cytotoxicity of its evaluated using MTT assay. The cell ELISA was used to determine the binding ability of immunotoxins (ITs) to the cell receptor and internalization and apoptosis were also assessed. The results revealed that cell cytotoxicity occurred in SKBR-3 cells in a dose-dependent manner but not in MCF-7 cells. It is possible that this ITs can attach to HER2-positive breast cancer cells and then, internalize and eradicate cancer cells by apoptosis. Here, we concluded that the recombinant ITs have therapeutic potential against HER2-positive breast cancer.

A new scfv-based recombinant immunotoxin against EPHA2-overexpressing breast cancer cells; High in vitro anti-cancer potency.

Immunotoxin therapy is one of the immunotherapy strategies providing a new, effective and high potency treatment against various cancers. Breast cancer is the most common cancer among women in many countries. The EPH receptors are a large part of tyrosine kinase receptors family and play an effective role in tumor development and angiogenesis. Among EPH receptors, EPHA2 is more commonly well-known and widely expressed in many cancers like breast cancer. In this study, we evaluated the specification of a designed immunotoxin formed by EPHA2-specific scfv linked with PE38KDEL on EPHA2-overexpressing breast cancer cell line. This new scfv-based recombinant immunotoxin was studied in terms of features such as binding potency, cytotoxicity effects, apoptosis induction ability, and internalization. The flow cytometry results showed that the immunotoxin can significantly (approximately 99%) bind to EPHA2-overexpressing breast cancer cell line (MDA-MB-231) in a low concentration (2.5 ng/ul) while cannot significantly bind to the normal cell line (HEK-293) or even EPHA2-very low expressing cell line (MCF-7). Using the MTT assay and Annexin V/Propidium iodide (PI) double staining method by flow cytometry, we observed significant killing and apoptosis induction of the MDA-MB-231 cells at different concentrations. Immunotoxin tracking by confocal microscopy at 2 h and 6 h revealed a massive presence of immunotoxin in the cytoplasm. Finally, given the in vitro results, it seems that this immunotoxin is competent enough to serve as a good candidate for in vivo studies to further explore the possibility of breast cancer treatment.

Satureja Khuzestanica Mediated Synthesis of Silver Nanoparticles and Its Evaluation of Antineoplastic Activity to Combat Colorectal Cancer Cell Line.

Several formulations of herbal plants have been extensively applied to treat diseases. Satureja khuzistanica (S. khuzistanica) is an Iranian traditional plant with a wide range of benefit effects on different diseases. In this study, we aimed to prepare silver nanoparticles from S. khuzistanica via the green synthesis method and investigate the anti-cancer effects on the HT29 cell line. To synthesize Ag-S. Khuzistanica, 50 mL S. khuzistanica extract and 1 mM AgNO3 were mixed and shaken at room temperature for 72 h. To determine the Ag-S. Khuzistanica nanoparticle characterization, XRD, FTIR, and TEM methods were done. In addition, MTT assay and real time PCR and annexin V/PI staining were performed to investigate the cytotoxicity, bcl-2 and bax gene expression and percentage of apoptotic cells. Our findings showed that Ag-S. khuzistanica is a spherical crystalline nanoparticle with the size less than 100 nm. MTT analysis showed that 375, 750, 1500 and 3000 µg/mL Ag-S. Khuzistanica significantly decreased the cell viability of HT29 cells. Ag-S. khuzistanica significantly reduced bcl-2 and increased apoptotic index expression at 375, 750, 1500, 3000 µg/mL Ag-S. Khuzistanica in a dose-dependent manner. Furthermore, cell staining with Annexin V/PI showed that treating Ag-S. Khuzistanica led to increasing in apoptotic cells. In conclusion, the formulation of Ag-S. khuzistanica has the apoptotic properties on the colorectal cancer cell line.

Micro RNA-126 promoting angiogenesis in diabetic heart by VEGF/Spred-1/Raf-1 pathway: effects of high-intensity interval training.

PURPOSE: This study aims to investigate the effect of high-intensity interval training (HIIT) on gene expression of MicroRNA-126 (miR-126) and serum concentration of vascular endothelial growth factor/ sprouty related EVH1 domain containing 1/ rapidly accelerated fibrosarcoma 1 (VEGF/Spred-1/Raf-1) proteins effective in cardiac tissue angiogenesis of diabetic rats. METHODS: Forty male Wistar rats were randomly divided into four groups of healthy control (HC), diabetic control (DC), diabetic with HIIT training (DT), and healthy with HIIT training (HT). HIIT was performed 6 days per week for 6 weeks (with the overload). Diabetes was induced via the combination of intraperitoneal injection of streptozotocin and high-fat foods. RESULTS: Diabetes remarkably diminished the expressions of miR-126, VEGF and Raf-1 proteins, and augmented Spred-1 expression. Meanwhile, the implementation of HIIT gave rise to a significant enhancement in expression of miR-126 heart tissue (P < 0.01), and subsequently increased the expression of VEGF and Raf-1 proteins (P < 0.01), and declined Spred-1 expression (P < 0.01) in the training group compared to the control group. CONCLUSION: The results of this study show that HIIT increases the expression of miR-126 by activating the angiogenesis pathway of the heart tissue. Increased angiogenesis through the miR-126 pathway is vital to compensate for heart destruction induced by diabetes. Thus, the use of standard interval exercise can be introduced as a novel therapeutic target for diabetic cardiomyopathy.

Characterization of Biosynthesized Silver Nanoparticles Using Lactobacillus rhamnosus GG and its In Vitro Assessment Against Colorectal Cancer Cells.

Silver nanoparticles are the most desirable nanoparticles broadly used in diverse fields. This study intends to investigate the anticancer properties of synthesized silver/Lactobacillus rhamnosus GG nanoparticles (Ag-LNPs) as a reducing and stabilizing agent in the synthesis process. To prepare silver/Lactobacillus rhamnosus GG nanoparticles, 1 mg/ml cell lysate of Lactobacillus rhamnosus GG and 1 mM silver nitrate solution were mixed and incubated for 72 h. XRD, FTIR, and TEM methods were used for nanoparticle characterization. MTT assay and annexin/PI staining were employed to analyze the toxicity and apoptotic cells levels of Ag-LNPs, respectively. TEM showed that these nanoparticles are spherical shaped about 233 nm in size. FTIR spectroscopy demonstrated that Ag-LNPs were functionalized with biomolecules. XRD pattern showed high purity and face-centered crystal structure of Ag-LNPs. MTT assay revealed that the percentages of HT-29 live cells significantly reduced in the high concentration of Ag-LNPs. Annexin/PI staining showed that these nanoparticles could lead HT-29 cells to apoptosis. This study showed the new Ag-LNP-synthesizing method using Lactobacillus rhamnosus GG as a cost-effective and efficient approach. Also, it showed that these nanoparticles can be considered as a potential active agent for biomedical applications and drug delivery due to their anticancer activities.

The crosstalk between trace elements with DNA damage response, repair, and oxidative stress in cancer.

DNA damage response (DDR) is a regulatory system responsible for maintaining genome integrity and stability, which can sense and transduce DNA damage signals. The severity of damage appears to determine DDRs, which can include damage repair, cell-cycle arrest, and apoptosis. Furthermore, defective components in DNA damage and repair machinery are an underlying cause for the development and progression of various types of cancers. Increasing evidence indicates that there is an association between trace elements and DDR/repair mechanisms. In fact, trace elements seem to affect mediators of DDR. Besides, it has been revealed that oxidative stress (OS) and trace elements are associated with cancer development. In this review, we discuss the role of some critical trace elements in the risk of cancer. In addition, we provide a brief introduction on DDR and OS in cancer. Finally, we will further review the interactions between some important trace elements including selenium, zinc, chromium, cadmium, and arsenic, and DDR, and OS in cancer.

Determining the effects of green chemistry synthesized Ag-nisin nanoparticle on macrophage cells.

Bacteriocins are low molecular weight substances produced through post transcriptional changes. These molecules are easily degraded in mammalian gut by proteolytic enzymes especially protease. Nisin is a peptide with 34 aa and its structure contains a pentacyclic lanthionine and 4 beta metyllanthionine residues. Different formulations have been designed for nisin. Since "green synthesis" is a progressive method to prepare anti-microbial and anti-cancer compounds, this study aimed at green synthesis of nisin metal compounds to be used lower concentration still exerting nisin effects. For this purpose, a 1 mg/ml nisin solution was added to a 1 mM silver nitrate solution and incubated to synthesis nano Ag-nisin, then the optical density of new solution was detected using UV spectroscopy. To determine biomolecules in the Ag-nisin solution, the FTIR method was employed. The size and morphology of Ag-nisin was measured by TEM. The toxicity, inflammatory cytokines production, and intracellular ROS quantity was evaluated using MTT, ELISA and flow-cytometry. XRD pattern indicated the silver crystals in Ag-nisin solution. In addition, FTRI findings showed that the carbonyl groups of amino acid are potently able to bind to metal nanoparticles, cover, and prevent them from particle agglomeration. Treating macrophage cells with 10, 25, 50 and 100 µg/ml of Ag-nisin had no significant effect on the cell viability and intracellular ROS quantity compared to the control group. In addition, different concentrations of Ag-nisin had no effect on the IL-10 and TNF-α levels but caused an increased level of IL-12 in comparison with the control group. In the current study, for the first time, green synthesize was used to prepare Ag-nisin particles. The synthesized nanoparticle is able to induce inflammatory activity via increasing IL-12 without any change in the TNF-α level in macrophage cells.

Evaluation of the antioxidant impact of ginger-based kombucha on the murine breast cancer model.

Background Abnormal metabolism is a common event in cancerous cells. For example, the increase of reactive oxygen species (ROS) production, particularly due to aerobic respiration during invasive stage, results in cancer progression. Herein, the impact of kombucha tea prepared from ginger on the alteration of antioxidant agents was assessed in the breast cancer animal model. Methods Two types of kombucha tea with or without ginger were administered to BALB/c mice before and after tumor challenge. Superoxide dismutase (SOD), catalase, glutathione (GSH) and malondialdehyde (MDA) were evaluated in tumor, liver and kidney. Results Administration of kombucha ginger tea significantly decreased catalase activity as well as GSH and MDA level in tumor homogenate (p<0.001). A significant decrease in SOD activity and increase in MDA quantity was determined in the kidney which had received kombucha ginger tea (p<0.01). Conclusions The consumption of kombucha prepared from ginger could exert minor antioxidant impacts by balancing multi antioxidant factors in different tissues in the breast cancer models.

The inhibitory impacts of Lactobacillus rhamnosus GG-derived extracellular vesicles on the growth of hepatic cancer cells.

Bacterial extracellular vesicles (EVs) have come forth into notice as possible important agent to mediate host-pathogen interactions. In this scientific research, the authors have tried to find out the effect of EVs derived from Lactobacillus rhamnosus GG (LDEVs) on the apoptosis induction in HepG2 cell line. The EVs were purified from the conditioned medium of Lactobacillus rhamnosus GG using ultrafiltration and confirmed by transmission electron microscopy (TEM). The HepG2 cells were treated with different concentrations of purified LDEVs and the cytotoxicity and their effects on the expression of bcl-2 and bax genes were assessed by the MTT assay and semi-quantitative RT-PCR, respectively. The MTT assay showed that only 100 µg/ml of LDEVs had a significant cytotoxic effect on cancer cells (p < 0.05). The apoptotic index (bax/bcl2 expression ratio) was significantly increased after treating with 50 and 100 µg/ml LDEVs (p < 0.05). Increased bax/bcl-2 ratio was led to cancer cell death.

Distribution of high-level mupirocin resistance among clinical MRSA.

Mupirocin is a topical antibiotic for methicillin-resistant Staphylococcus aureus (MRSA) decolonization in hospital settings and nursing homes and is used as a highly effective antibiotic against MRSA. In this study, we aimed to evaluate the frequency of high-level mupirocin-resistant (HLMR) strains among the MRSA subtypes. A total of 188 clinical MRSA isolates were collected from 2011 to 2014, and their susceptibility to antimicrobial agents and vancomycin resistance was evaluated using disc diffusion method and micro-dilution method, respectively. Furthermore, the presence of mecA, SSCmec, mupA and mupB was assessed by PCR. All isolates were multi-drug resistant (MDR) but 2 strains (1.06%) were resistant to mupirocin. Minimum inhibitory concentration (MIC) of vancomycin for 8 strains (4.7%) was higher than 2 µg/ml. Of 188 isolates, 188 (100%), 64 (34.04%), 8 (4.3%), 150 (79.8%), 26 (13.8%), 2 (1.06) and 2 (1.06%) isolates possessed mecA, SCCmec types I, II, III, IV, mupA and mupB genes, respectively. Our data showed that despite infection control policy enforced by health care committee, the rate of mupirocin resistance among MRSA strains is continuously rising.