Production of biosurfactant by salt-resistant Bacillus in lead-supplemented media: application and toxicity / Producción de biosurfactante por Bacillus resistente a la sal en medios suplementados con plomo: aplicación y toxicidad

A group of biosurfactants are lipopeptides that are produced by some microorganisms, especially Bacillus strains. They are new bioactive agents with anticancer, antibacterial, antifungal, and antiviral activities. Also, they are used in sanitation industries. In this study, a lead-resistant strain of Bacillus halotolerans was isolated for lipopeptide production. This isolate exhibited metal resistance (lead, calcium, chromium, nickel, copper, manganese, and mercury), salt tolerance (12%), and antimicrobial activities against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Saccharomyces cerevisiae. The production of lipopeptide was optimized, concentrated, and then extracted from the polyacrylamide gel in a simple way for the first time. The nature of the purified lipopeptide was determined by FTIR, GC/MS, and HPLC analyses. The purified lipopeptide indicated significant antioxidant properties (90.38% at a concentration of 0.8 mg ml−1). Also, it had anticancer activity by apoptosis (flow cytometry analysis) in MCF-7 cells, while it had no cytotoxicity on HEK-293 normal cells. Therefore, Bacillus halotolerans lipopeptide has the potential to be used as an antioxidant, antimicrobial, or anticancer agent in the medical and food industries.(AU)

The potential effect of silver nanoparticles synthesized with Coffea arabica green seeds on Leishmania major proliferation, cytotoxicity activity, and cytokines expression level.

The goal of this study was to analyze the antileishmanial and antibacterial activity of Coffea arabica green seed biosynthesize silver nanoparticles (C. arabica AgNPs), as well as cytotoxicity and cytokine gene expression. UV-vis spectroscopy, FTIR, and FESEM methods used to examine the C. arabica AgNPs. MTT test was used to assess the antileishmanial and cytotoxicity effects. The gene expression level was assessed in NPs-treated J774 cells by qPCR. The synthesized C. arabica AgNPs were in the size range of 20-70 nm, through FESEM pictures. The IC50 values of the NPs were 65. 4 and 47.70 µg/mL against promastigotes and amastigotes of Leishmania major, but these values were 580.1 and 171.1 µg/mL for Glucantime® as the control drug. C. arabica AgNPs represented a significant increase in IL-12P40, as a Th1 cytokine, in comparison to Glucantime® at high concentrations (P < 0.01), whilst IL-10 expression level showed a significant reduction between NPs-treated and Glucantime®-treated macrophages at 250-1000 µg/mL concentrations (P < 0.001). Moreover, the NPs were cytotoxic on cancer cell lines of Hek293, MCF7, and A172 with the CC50 values of 437.2, 116.8, and 72.9 µg/mL, respectively. It showed a significant effect of these NPs against A172 (P < 0.001). Also, the lowest MIC values of the NPs were obtained for Bacillus subtilis and Staphylococcus aureus (204 µg/mL). According to the antileishmanial, anticancer, and antibacterial activity of these NPs, it can considered a bio-agent drug in the future in endemic countries.

Production of biosurfactant by salt-resistant Bacillus in lead-supplemented media: application and toxicity.

A group of biosurfactants are lipopeptides that are produced by some microorganisms, especially Bacillus strains. They are new bioactive agents with anticancer, antibacterial, antifungal, and antiviral activities. Also, they are used in sanitation industries. In this study, a lead-resistant strain of Bacillus halotolerans was isolated for lipopeptide production. This isolate exhibited metal resistance (lead, calcium, chromium, nickel, copper, manganese, and mercury), salt tolerance (12%), and antimicrobial activities against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Saccharomyces cerevisiae. The production of lipopeptide was optimized, concentrated, and then extracted from the polyacrylamide gel in a simple way for the first time. The nature of the purified lipopeptide was determined by FTIR, GC/MS, and HPLC analyses. The purified lipopeptide indicated significant antioxidant properties (90.38% at a concentration of 0.8 mg ml-1). Also, it had anticancer activity by apoptosis (flow cytometry analysis) in MCF-7 cells, while it had no cytotoxicity on HEK-293 normal cells. Therefore, Bacillus halotolerans lipopeptide has the potential to be used as an antioxidant, antimicrobial, or anticancer agent in the medical and food industries.

Thermal and Microwave-Assisted Synthesis of New Highly Functionalized Bis-ß-lactams from Available Compounds via Bisketene as an Intermediate.

The synthesis of highly functionalized bis-ß-lactams containing aromatic rings was achieved by thermal and microwave-assisted methods starting from easily available 2-(4-hydroxyphenyl)acetic acid and 2,2'-(propane-2,2-diyl)diphenol precursors. The approach to these valuable heterocyclic scaffolds involved formal [2π + 2π] cycloadditions between Schiff bases and novel bisketenes, which were generated in situ, followed by an electrocyclic reaction of zwitterionic intermediates. Reactions carried out under microwave irradiation were clean and gave high yields with significantly reduced reaction times. Interestingly, in the thermal method, the reaction proceeded in a stereospecific manner, and only the trans-cis or cis-cis isomers were formed. However, under the microwave conditions, the reaction proceeded stereoselectively, and other possible isomers such trans-trans and cis-trans isomers were formed in addition to the product formed under thermal conditions. More interestingly, when the two compounds that did not produce any products under thermal conditions were reacted under microwave conditions, one formed the trans-cis isomer and the other formed the cis-trans and trans-trans isomers as two products .

Characteristics and in vitro anti skin aging activity and UV radiation protection of morin loaded in niosomes.

BACKGROUND: One of the dermatologic problems in elderly people is skin aging, which is a natural and complex biological process. Morin is a flavonoid with high radical scavenging activity as well as antityrosinase effects but its low water solubility has restricted its application. AIMS: This research aimed to develop, characterize, and optimize morin niosomes composed of non-ionic surfactants, and evaluate the in vitro UV protection and antiaging effectiveness. METHODS: Niosomes were prepared by the film hydration method with sorbitan monostearate (Span® 40), polyoxyethylenesorbitan monopalmitate (Tween® 40), and cholesterol. The niosomes were characterized in terms of size, zeta potential, morphology, in vitro release behavior, and drug entrapment efficiency (EE). Afterward, antiaging activity, including antityrosinase, antioxidant, intracellular reactive oxygen species (ROS) scavenging, and sun protection factor (SPF) were evaluated. RESULTS: The optimized niosomes appeared as unilamellar vesicles with a spherical shape, with size, zeta potential, and EE values of 6.13 ± 0.40 µm, -0.81 ± 0.32 mV, and 89.35% ± 2.80%, respectively. The noisome formulation remained stable at -4°C for 3 months. The release profiles of morin loaded in niosomes revealed the extended release over 8 h and followed zero-order release kinetics. Morin-loaded niosomes exhibited no significant toxicity toward the L929 cell line. The niosome loaded with morin showed anti skin aging activity, including antityrosinase effects (IC50 = 13.17 ± 1.58 µg/ml), antioxidant (IC50 = 28.49 ± 2.05 µg/ml), and ROS scavenging activity. For 1% and 5% (w/w) morin niosomes in eucerin base cream, the SPF was 39.03 ± 1.01 and 38.15 ± 0.82, respectively, whereas the noisome-free morin cream exhibited an SPF of 4.47 ± 0.56. CONCLUSION: Morin-loaded niosome has been shown to provide sun protection and antiaging effects, suggesting that it could be used in pharmaceutical and cosmetic products.

Chemical composition, anticancer and antibacterial activity of Nepeta mahanensis essential oil.

BACKGROUND: Conventional cancer treatments, such as chemotherapy, radiation therapy, and surgery, often affect the patients' quality of life due to their serious side effects, indicating the urgent need to develop less toxic and more effective alternative treatments. Medicinal plants and their derivatives are invaluable sources for such remedies. The present study aimed to determine the chemical composition, anticancer and antibacterial activities of Nepeta mahanesis essential oil (EO). METHODS: The chemical composition of EO was analyzed by gas chromatography-mass spectrometry (GC-MS). Cytotoxicity and apoptosis/necrosis induction of EO was analyzed by MTT assay and Flow cytometry. Real-time PCR was performed to evaluate the Bax/Bcl2 gene expression. Also, the effect of the EO on the cells' mitochondrial membrane potential (MMP) and ROS level was assessed. DPPH assay was done to assess the free radical scavenging activity of the EO. The Antimicrobial activity, MIC, and MBC of the oil were determined via well-diffusion and broth microdilution methods. RESULTS: Based on the GC-MS analysis, 24 compounds were identified in the EO, of which 1,8-cineole (28.5%), Nepetalactone (18.8%), germacrene D (8.1%), and ß-pinene (7.2%), were the major compounds. Also, the EO showed considerable cytotoxicity against MCF-7, Caco-2, SH-SY5Y, and HepG2 after 24 and 48 h treatment with IC50 values between 0.0.47 to 0.81 mg/mL. It was revealed that this compound increased the Bax/Bcl2 ratio in the MCF-7 cells and induced apoptosis (27%) and necrosis (18%) in the cells. Moreover, the EO treatment led to a substantial decrease in MMP, which is indicative of apoptosis induction. A significant increase in ROS level was also detected in the cells following exposure to the EO. This compound showed strong DPPH radical scavenging activity (IC50: 30). It was also effective against Gram-positive E. faecalis (ATCC 29,212) and Gram-negative E. coli (ATCC 11,333) bacteria. CONCLUSIONS: The results of this study demonstrated that the EO of N. mahanesis could be considered a bioactive product with biomedical applications that can be used as an alternative cancer treatment and applied in the biomedical industries.

Borderline Boolean states improve the biosensing applications of DNA circuits.

Molecular circuits have been used in a wide range of diagnosis applications, from the detection of chemical molecules in solution to the complex processing of cell surface receptors. One of the most important challenges of these systems is the lack of distinguishability between different circuit states when each circuit state represents a specific disease. In this work, we designed a molecular amplification circuit with borderline Boolean states that each state can be distinguished with different color intensity. For this purpose, two DNA complexes and four DNA hairpin structures were designed to detect miR-218 and miR-215 biomarkers. One of the designed DNA complexes has two G-quadruplex structures and the other has only one G-quadruplex structure. In the absence of the inputs, all three G-quadruplex structures are active and produce a high-intensity signal, while in the other three states, including the presence of miR-218, the presence of miR-215, and the presence of both inputs, respectively, one, two, and zero G-quadruplex structures are active. Therefore, the designed system can identify two different biomarkers simultaneously with different signal ratios, which can easily distinguish the different states of the circuit. This strategy is very promising to identify diseases in which any combination of biomarkers leads to a particular disease.

Effect of Menadione and Combination of Gemcitabine and Cisplatin on Cancer Stem Cells in Human Non-small Cell Lung Cancer (NSCLC) Cell Line A549.

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. Chemotherapy-induced adverse effects and resistance of NSCLC to conventional drugs reduce the efficacy of current therapies. Tumors contain a small population of cancer stem cells (CSCs) that play a critical role in tumor initiation, maintenance, and drug resistance that finally lead to cancer recurrence. Therefore, CSC-targeting therapies can offer the best hope for developing curative cancer therapies. Vitamins have a high potential for cancer prevention and treatment. Vitamins also ameliorate the side effects which occur in chemo-radio therapy. Menadione (2-methyl-1,4-naphthoquinone/vitamin-K3) is a synthetic form of vitamin K that indicated antitumor activities. The purpose of this study was to evaluate the anti-CSCs effect of menadione and combination of cisplatin and gemcitabine as a first-line treatment in patients with NSCLC on the NSCLC cell line A549. MTT results displayed decreased cell survival after treatment with cisplatin/gemcitabine for 48 h treatment (IC50 values 0.25 µM for cisplatin and 5 µM for gemcitabine). Menadione also inhibited the cell growth in A549 cells (IC50: 16 µM). Quantitative RT-PCR showed significant downregulation of CSC markers (Oct4, Nanog, Sox2, Aldh1, Abcb1, CD44, and CD133) and Snail, epithelial-mesenchymal transition marker, after treatment with menadione and cisplatin/gemcitabine. Flow cytometry showed CD44-positive cells that constitute a high percentage (70%) of A549 cells reduced significantly after treatment with cisplatin/gemcitabine or menadione. However, A549 cells did not show a significant population positive for CD133 and ABCB1 (less than 0.05%), and these fractions did not change after treatment with two agents.

Phytol Down-Regulates Expression of Some Cancer Stem Cell Markers and Decreases Side Population Proportion in Human Embryonic Carcinoma NCCIT Cells.

Cancer stem cells (CSCs), a subgroup of cancer cells, have self-renewal capacity and differentiation potential and drive tumor growth. CSCs are highly-resistant to conventional chemo-radio therapy. Phytochemicals were shown to be able to eliminate CSCs. Phytol is a diterpene alcohol with demonstrated anticancer effects. The current study compared the effect of phytol with retinoic acid (RA) as a well-known inducers of CSC differentiation and cisplatin, a common chemotherapy drug, on CSC markers in human embryonic carcinoma NCCIT cells. NCCIT cells were exposed to 10 mM RA for 14 day to induce differentiation. Moreover, NCCIT cells were treated with IC50 dose of cisplatin (12 µM) and phytol (40 µM) for 7 day. Real-time PCR showed that phytol was more effective that RA and cisplatin in down-regulating the CSC markers OCT4, NANOG, SOX2, ALDH1, ABCB1, CD44 and CD133. Percentage of SP (13%) and ABCB1+ (0.34%) in NCCIT cells decreased to 7% and 0.1% respectively after treatment with phytol. A very small proportion of NCCIT cells were positive for CD44 (0.2%) and CD133 (0.48%) and this fraction did not change significantly after treatment with three agents. In conclusion, phytol has the greatest inhibitory effect on CSC population and markers than RA and cisplatin.

Evaluation of anticancer, antioxidant and antibacterial properties of methanol extract of three Acantholimon Boiss. species.

OBJECTIVE: Acantholimon is a genus of perennial plant within the Plumbaginaceae family. Here, we aimed to investigate anticancer, antioxidant, and antibacterial potential of methanol extract of three Iranian endemic species of Acantholimon including A. austro-iranicum, A. serotinum and A. chlorostegium. MATERIALS AND METHODS: MTT assay was used to evaluate the in vitro cytotoxicity and apoptosis induction was examined by annexin V-PE apoptosis detection kit. Antioxidant activity was reported based on the DPPH-scavenging and DCF-DA assay. Antibacterial activity was measured by disc diffusion and micro-well dilution assay. RESULTS: MTT assay showed less cytotoxicity of methanol extracts against the HUVEC normal cell line (IC50 values: 817-900 µg/ml) compared to cancer cell lines MCF-7, HT29, SH-SY5Y, NCCIT and A549 (IC50 values: 213 to 600 µg/ml) that show the specificity of extracts toward cancer cells. Plant extract showed apoptosis induction and cell cycle arrest at the G0/G1 phases documented by annexin V staining and flow cytometry. According to antioxidant tests, extracts exhibited significant DPPH scavenging potential (IC50 values: 30-37 µg/ml) and could protect against H2O2-induced oxidative stress. Antibacterial activities showed a stronger inhibitory effect on Escherichia coli and Pseudomonas aeruginosa as Gram- negative bacteria (diameter of inhibition zone: 11-13 mm and minimal inhibition concentration (MIC): 3.175 to 12.5 mg/ml) compared to Gram-positive bacteria including Enterococcus faecalis and Staphylococcus aureus (diameter of inhibition zone: 3-7 mm and MIC: 25 to 50 mg/ml). CONCLUSION: Our results suggested moderate cytotoxic and antibacterial potential and noteworthy antioxidant activity for the examined Acantholimon species.

Data on analysis of OCC-1 transcript levels in pluripotent and differentiated states of P19 cells.

We investigated the expression of OCC-1 at mRNA level during retinoic acid (RA) induced differentiation of mouse embryonic carcinoma P19 pluripotent cancer cells by quantitative real time PCR (qPCR). By employing four-fold serial dilutions of P19 cDNA, standard curves were generated for the reference gene (L37) and the gene of interest (OCC-1). PCR efficiencies for L37 and OCC-1 were calculated. Since the amplification efficiencies of these two genes were unequal, the standard curve method was used for the relative quantification of OCC-1. Data analysis revealed that the expression of OCC-1 was reduced by about 69% after 4-day treatment with RA, when significant down-regulation of key pluripotency factors, including OCT4 and Nanog was observed [1].

Phytochemical composition, and cytotoxic, antioxidant, and antibacterial activity of the essential oil and methanol extract of Semenovia suffruticosa.

OBJECTIVE: In this study, our aim was to extract, and identify and quantify the chemical composition of essential oils of Semenovia suffruticosa grown in Kerman, Iran. Moreover, cytotoxic, antioxidant and antimicrobial activity of the essential oil and methanol extract of aerial parts of S. suffruticosa were reported. MATERIALS AND METHODS: GC and GC/MS analysis were used for identifying and quantifying the essential oil components. Antioxidant and antibacterial activity were tested by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and agar disc diffusion methods, respectively and MTT assay was used to determine the anti-proliferative potential of the oil against breast (MCF-7), colon (HT-29), neuroblastoma (SH-SY5Y), embryonal carcinoma (NCCIT) cancer cell relative to human umbilical vein endothelial cell (HUVEC) as a normal cell. Apoptosis induction was monitored by flow cytometry using PE annexin V apoptosis detection kit and cell cycle arrest was by with propidium iodide. RESULTS: Z-ß-ocimene (25.1%), linalool (17.8%) and ß-bisabolol (13.3%) were recognized as major components of the essential oil. Our study demonstrated apoptosis-inducing potential of essential oil on normal and cancer cells. However, methanol extract exerted cytotoxicity against a number of cancer cells and arrested cancer cells in G2/M phase; nevertheless, it did not exert strong cytotoxicity against normal cells. Furthermore, DPPH and disc diffusion results showed that while essential oil has considerable antiradical activity, methanol extract did not exert promising antioxidant and antimicrobial activity. CONCLUSION: Methanol extract of S. suffruticosa shows tumor-cell-specific cytotoxic properties and the essential oil demonstrated a strong antioxidant activity.

BORIS: a key regulator of cancer stemness.

BORIS (CTCFL) is a DNA binding protein which is involved in tumorigenesis. Although, there are different opinions on the level of gene expression and function of BORIS in normal and cancer tissues, the results of many studies have classified BORIS as a protein belonging to cancer/testis (CT) genes, which are identified as a group of genes that are expressed normally in testis, and abnormally in various types of cancers. In testis, BORIS induces the expression of some male germ cell/testis specific genes, and plays crucial roles during spermatogenesis and production of sperm. In tumorigenesis, the role of BORIS in the expression induction of some CT genes and oncogenes, as well as increasing proliferation/viability of cancer cells has been demonstrated in many researches. In addition to cancer cells, some believe that BORIS is also expressed in normal conditions and plays a universal function in cell division and regulation of genes. The following is a comprehensive review on contradictory views on the expression pattern and biological function of BORIS in normal, as well as cancer cells/tissues, and presents some evidence that support the expression of BORIS in cancer stem cells (CSCs) and advanced stage/poorer differentiation grade of cancers. Boris is involved in the regulation of CSC cellular and molecular features such as self-renewal, chemo-resistance, tumorigenicity, sphere-forming ability, and migration capacity. Finally, the role of BORIS in regulating two important signaling pathways including Wnt/ß-catenin and Notch in CSCs, and its ability in recruiting transcription factors or chromatin-remodeling proteins to induce tumorigenesis is discussed.

Effect of Cinnamic acid and FOLFOX in diminishing side population and downregulating cancer stem cell markers in colon cancer cell line HT-29.

PURPOSE: There is a lot of evidence suggesting that a small subset of cancer cells resistant to conventional chemotherapy and radiotherapy and known as cancer stem cells (CSCs) is responsible for promoting metastasis and cancer relapse. Therefore, targeting and eliminating the CSCs could lead to higher survival rates and a better quality of life. In comparison with conventional chemical drugs that may not be effective against CSCs, phytochemicals are strong anti-CSCs agents. The current study examines the effect of 5-fluorouracil plus oxaliplatin (FOLFOX) as a common chemotherapy drug on colorectal cancer as well as the influence of Cinnamic acid (CINN) as a plant-derived phytochemical on colon cancer stem-like cells in HT-29 adenocarcinoma cell line. METHODS: The anti-proliferative effect of FOLFOX and CINN was determined using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Flow cytometry analysis was used for the identification of side population (SP), CD44, and CD133 positive cells. The expression of OCT4, NANOG, ABCB1, and ALDH1A was assessed by RT-PCR. RESULTS: The FOLFOX and CINN decreased cell viability in certain drug concentrations: IC50 = 5,40 µM oxaliplatin +220 µM 5-fluorouracil, and 13,50 mM for CINN. The CSC-associated markers (OCT4, NANOG, ABCB1, and ALDH1A) and the proportion of cancer stem-like cells (SP cells, CD44, and CD133 positive cells) were downregulated following the treatment of HT-29 adenocarcinoma cell line with IC50 concentrations of FOLFOX and CINN. CONCLUSION: Our data suggests that CINN, a naturally occurring component, could be more effective than FOLFOX treatment in reducing the cancer stem-like cells and expression of CSC markers from HT-29 colon cancer cells. Graphical abstract ᅟ.

Perspective: Cooperation of Nanog, NF-κΒ, and CXCR4 in a regulatory network for directed migration of cancer stem cells.

Directed cell migration is a crucial mobility phase of cancer stem cells having stemness and tumorigenic characteristics. It is known that CXCR4 plays key roles in the perception of chemotactic gradients throughout the directed migration of CSCs. There are a number of complex signaling pathways and transcription factors that coordinate with CXCR4/CXCL12 axis during directed migration. In this review, we focus on some transcription factors such as Nanog, NF-κB, and Bmi-1 that cooperate with CXCR4/CXCL12 for the maintenance of stemness and induction of metastasis behavior in cancer stem cells.

Expression analysis of BORIS during pluripotent, differentiated, cancerous, and non-cancerous cell states.

BORIS/CTCFL is an 11 zinc finger protein, which is the paralog of CTCF, a ubiquitously expressed protein with diverse roles in gene expression and chromatin organization. Several studies have shown that the expression of BORIS is restricted to normal adult testis, pluripotent cells, and diverse cancer cell lines. Thus, it is known as a cancer-testis (CT) gene that has been hypothesized to exhibit oncogenic properties and to be involved in cancer cell proliferation. On the contrary, other reports have shown that its expression is more widespread and can be detected in differentiated and normal somatic cells; hence, it might have roles in general cellular functions. The present study was aimed to analyze the expression of BORIS in different cell states of pluripotent, differentiated, cancerous and non-cancerous.We found that the two cell states of pluripotency and differentiation are not accompanied with significant variations of BORIS expression. Furthermore, Boris transcripts were detected at approximately the same level in cancer and non-cancer cell lines. These findings suggest that, in contrast to some previous reports, the expression of mouse BORIS is not limited to only cancerous cells or pluripotent cell states.

Cancer stem cells and cancer therapy.

Cancer stem cells (CSCs) are a subpopulation of tumour cells that possess the stem cell properties of self-renewal and differentiation. Stem cells might be the target cells responsible for malignant transformation, and tumour formation may be a disorder of stem cell self-renewal pathway. Epigenetic alterations and mutations of genes involved in signal transmissions may promote the formation of CSCs. These cells have been identified in many solid tumours including breast, brain, lung, prostate, testis, ovary, colon, skin, liver, and also in acute myeloid leukaemia. The CSC theory clarifies not only the issue of tumour initiation, development, metastasis and relapse, but also the ineffectiveness of conventional cancer therapies. Treatments directed against the bulk of the cancer cells may produce striking responses but they are unlikely to result in long-term remissions if the rare CSCs are not targeted. In this review, we consider the properties of CSCs and possible strategies for controlling the viability and tumourigenecity of these cells, including therapeutic models for selective elimination of CSCs and induction of their proper differentiation.