27-Hydroxycholesterol represses G9a expression via oestrogen receptor alpha in breast cancer.

27-hydroxycholesterol (27-HC) is a cholesterol metabolite and the first discovered endogenous selective estrogen receptor modulator (SERM) that has been shown to have proliferative and metastatic activity in breast cancer. However, whether 27-HC metabolite modulates the epigenetic signatures in breast cancer and its progression remains unclear. The current study, reports that 27-HC represses the expression of euchromatic histone lysine methyltransferase G9a, further reducing di-methylation at H3K9 in a subset of genes. We also observed reduced occupancy of ERα at the G9a promoter, indicating that 27-HC negatively regulates the ERα occupancy on the G9a promoter and functions as a transcriptional repressor. Further, ChIP-sequencing for the H3K9me2 mark has demonstrated that 27-HC treatment reduces the H3K9me2 mark on subset of genes linked to cancer progression, proliferation, and metastasis. We observed upregulation of these genes following 27-HC treatment which further confirms the loss of methylation at these genes. Immunohistochemical analysis with breast cancer patient tissues indicated a positive correlation between G9a expression and CYP7B1, a key enzyme of 27-HC catabolism. Overall, this study reports that 27-HC represses G9a expression via ERα and reduces the levels of H3K9me2 on a subset of genes, including the genes that aid in breast tumorigenesis and invasion further, increasing its expression in the breast cancer cells.

Urolithin A: A promising selective estrogen receptor modulator and 27-hydroxycholesterol attenuator in breast cancer.

27-hydroxycholesterol (27-HC) is an oxysterol that acts as an endogenous selective estrogen receptor modulator (SERM), and its adverse effects on breast cancer via the estrogen receptor (ER) have provided new insights into the pathology of cholesterol-linked breast cancer. Our earlier in vitro experiments showed that the methanolic extract of pomegranate could exhibit SERM properties and compete with 27-HC. The major constituents of pomegranate are ellagitannins and ellagic acid, which are converted into urolithins by the colonic microbiota. In recent years, urolithins, especially urolithin A (UA) and urolithin B (UB), have been reported to have a plethora of advantageous effects, including antiproliferative and estrogenic activities. In this study, we attempted to determine the potential of urolithins in antagonizing and counteracting the adverse effects of 27-HC in breast cancer cells. Our findings suggested that UA had an antiproliferative capacity and attenuated the proliferative effects of 27-HC, resulting in subsequent loss of membrane potential and apoptosis in breast cancer cells. Further, UA induced estrogen response element (ERE) transcriptional activity and modulated estrogen-responsive genes, exhibiting a SERM-like response concerning receptor binding. Our in vivo hollow fiber assay results showed a loss of cell viability in breast cancer cells upon UA consumption, as well as a reduction in 27-HC-induced proliferative activity. Additionally, it was shown that UA did not induce uterine proliferation or alter blood biochemical parameters. Based on these findings, we can conclude that UA has the potential to act as a potent estrogen receptor alpha (ERα) modulator and 27-HC antagonist. UA is safe to consume and is very well tolerated. This study further opens up the potential of UA as ER modulator and its benefits in estrogen-dependent tissues.

RXR agonist, Bexarotene, effectively reduces drug resistance via regulation of RFX1 in embryonic carcinoma cells.

Aberrant expression of multidrug resistance (MDR) proteins is one of the features of cancer stem cells (CSCs) that make them escape chemotherapy. A well-orchestrated regulation of multiple MDRs by different transcription factors in cancer cells confers this drug resistance. An in silico analysis of the major MDR genes revealed a possible regulation by RFX1 and Nrf2. Previous reports also noted that Nrf2 is a positive regulator of MDR genes in NT2 cells. But we, for the first time, report that Regulatory factor X1 (RFX1), a pleiotropic transcription factor, negatively regulates the major MDR genes, Abcg2, Abcb1, Abcc1, and Abcc2, in NT2 cells. The levels of RFX1 in undifferentiated NT2 cells were found to be very low, which significantly increased upon RA-induced differentiation. Ectopic expression of RFX1 reduced the levels of transcripts corresponding to MDRs and stemness-associated genes. Interestingly, Bexarotene, an RXR agonist that acts as an inhibitor of Nrf2-ARE signaling, could increase the transcription of RFX1. Further analysis revealed that the RFX1 promoter has binding sites for RXRα, and upon Bexarotene exposure RXRα could bind and activate the RFX1 promoter. Bexarotene, alone or in combination with Cisplatin, could inhibit many cancer/CSC-associated properties in NT2 cells. Also, it significantly reduced the expression of drug resistance proteins and made the cells sensitive towards Cisplatin. Our study proves that RFX1 could be a potent molecule to target MDRs, and Bexarotene can induce RXRα-mediated RFX1 expression, therefore, would be a better chemo-assisting drug during therapy.

Early Detection of Biofouling on Water Purification Membranes by Ambient Ionization Mass Spectrometry Imaging.

By direct analysis of water purification membranes using ambient ionization mass spectrometry, an attempt has been made to understand the molecular signatures of bacterial fouling. Membrane based purification methods are used extensively in water treatment, and a major challenge for them is biofouling. The buildup of microbes and their extracellular polymeric matrix clog the purification membranes and reduce their efficiency. To understand the early stages of bacterial fouling on water purification membranes, we have used desorption electrospray ionization mass spectrometry (DESI MS), where ion formation occurs in ambient conditions and the ionization event is surface sensitive. Biosurfactants at the air-water interface generated by microorganisms as a result of quorum sensing, influence the water-membrane interface and are important for the bacterial attachment. We show that these biosurfactants produced by bacteria can be indicator molecular species signifying initiation of biofilms on membrane surfaces, demonstrated by specific DESI MS signatures. In Pseudomonas aeruginosa, one of the best studied models for biofilm formation, this process is mediated by rhamnolipids forewarning bacterial fouling. Species dependent variation of such molecules can be used for the precise identification of the microorganisms, as revealed by studies on P. aeroginosa (ATCC 25619). The production of biosurfactants is tightly regulated at the transcriptional level by the quorum-sensing (QS) response. Thus, secretion of these extracellular molecules across the membrane surface allows rapid screening of the biofilm community. We show that, the ambient ionization mass spectrometry can detect certain toxic heavy metals present in water, using surfactant-metal complexes as analytes. We believe that such studies conducted on membranes in various input water streams will help design suitable membrane processes specific to the input streams.

Holey MoS2 Nanosheets with Photocatalytic Metal Rich Edges by Ambient Electrospray Deposition for Solar Water Disinfection.

A new method for creating nanopores in single-layer molybdenum disulfide (MoS2) nanosheets (NSs) by the electrospray deposition of silver ions on a water suspension of the former is introduced. Electrospray-deposited silver ions react with the MoS2 NSs at the liquid-air interface, resulting in Ag2S nanoparticles which enter the solution, leaving the NSs with holes of 3-5 nm diameter. Specific reaction with the S of MoS2 NSs leads to Mo-rich edges. Such Mo-rich defects are highly efficient for the generation of active oxygen species such as H2O2 under visible light which causes efficient disinfection of water. 105 times higher efficiency in disinfection for the holey MoS2 NSs in comparison to normal MoS2 NSs is shown. Experiments are performed with multiple bacterial strains and a virus strain, demonstrating the utility of the method for practical applications. A conceptual prototype is also presented.

Organic Solvent-Free Fabrication of Durable and Multifunctional Superhydrophobic Paper from Waterborne Fluorinated Cellulose Nanofiber Building Blocks.

In view of a great demand for paper-based technologies, nonwettable fibrous substrates with excellent durability have drawn much attention in recent years. In this context, the use of cellulose nanofibers (CNFs), the smallest unit of cellulosic substrates (5-20 nm wide and 500 nm to several microns in length), to design waterproof paper can be an economical and smart approach. In this study, an eco-friendly and facile methodology to develop a multifunctional waterproof paper via the fabrication of fluoroalkyl functionalized CNFs in the aqueous medium is presented. This strategy avoids the need for organic solvents, thereby minimizing cost as well as reducing safety and environmental concerns. Besides, it widens the applicability of such materials as nanocellulose-based aqueous coatings on hard and soft substrates including paper, in large areas. Water droplets showed a contact angle of 160° (±2°) over these surfaces and rolled off easily. While native CNFs are extremely hydrophilic and can be dispersed in water easily, these waterborne fluorinated CNFs allow the fabrication of a superhydrophobic film that does not redisperse upon submersion in water. Incorporated chemical functionalities provide excellent durability toward mechanochemical damages of relevance to daily use such as knife scratch, sand abrasion, spillage of organic solvents, etc. Mechanical flexibility of the chemically modified CNF composed paper remains intact despite its enhanced mechanical strength, without additives. Superhydrophobicity induced excellent microbial resistance of the waterproof paper which expands its utility in various paper-based technologies. This includes waterproof electronics, currency, books, etc., where the integrity of the fibers, as demonstrated here, is a much-needed criterion.