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Transcriptional control has been earnestly pursued for the regulation of cellular proliferation associated with cancer progression. The foundational paradigm of targeting transcription factors has yielded exquisite specificity, but many factors cannot yet be targeted. In contrast, targeting epigenetic factors to control chromatin structure and consequential gene expression generally yields more global effects on transcription. Our working paradigm targets neither specific transcription factors nor global epigenetic factors but ATP-dependent chromatin remodeling factors that regulate expression of a limited set of genes. Active DNA-dependent ATPase A Domain inhibitor (ADAADi) synthesized by aminoglycoside phosphotransferases is the first-in-class inhibitor of ATP-dependent chromatin remodeling proteins that targets the ATPase domain of these proteins. Mammalian cells are sensitive to ADAADi but cell lines are variable in their individual responses to the inhibitor. The ADAADi product can be generated from a variety of aminoglycoside substrates with cells showing differential responses to ADAADi depending on the starting aminoglycoside. RNA seq analysis demonstrated that targeting the chromatin remodeling by treatment with a sub-lethal concentration of ADAADi yields alterations to the transcriptional network of the cell. Predominantly, the tumor-promoting genes were repressed while pro-apoptotic and tumor suppressors genes were upregulated on treatment with ADAADi, leading to apoptotic-type cell death. Treatment with ADAADi reversed the EMT process as well as inhibited migration of cells and their colony forming ability. In conjunction with the previous report that treatment with ADAADi regresses tumors in mouse model, this chromatin remodeling inhibitor shows promising anti-tumor properties by targeting the main hallmarks of cancer.

Active DNA-dependent ATPase A Domain inhibitor (ADAADi) is the only known inhibitor of ATP-dependent chromatin remodeling proteins that targets the ATPase domain of these proteins. The molecule is synthesized by aminoglycoside phosphotransferase enzyme in the presence of aminoglycosides. ADAADi interacts with ATP-dependent chromatin remodeling proteins through motif Ia present in the conserved helicase domain, and thus, can potentially inhibit all members of this family of proteins. We show that mammalian cells are sensitive to ADAADi but with variable responses in different cell lines. ADAADi can be generated from a wide variety of aminoglycosides; however, cells showed differential response to ADAADi generated from various aminoglycosides. Using HeLa and DU145 cells as model system we have explored the effect of ADAADi on cellular functions. We show that the transcriptional network of a cell type is altered when treated with sub-lethal concentration of ADAADi. Although ADAADi has no known effects on DNA chemical and structural integrity, expression of DNA-damage response genes was altered. The transcripts encoding for the pro-apoptotic proteins were found to be upregulated while the anti-apoptotic genes were found to be downregulated. This was accompanied by increased apoptosis leading us to hypothesize that the ADAADi treatment promotes apoptotic-type of cell death by upregulating the transcription of pro-apoptotic genes. ADAADi also inhibited migration of cells as well as their colony forming ability leading us to conclude that the compound has effective anti-tumor properties.

// Qiong Wu 1 , Pasil Madany 1 , Jason R. Dobson 1 , Jake M. Schnabl 1 , Soni Sharma 2 , Tara C. Smith 1 Andre J. van Wijnen 3 , Janet L. Stein 4 , Jane B. Lian 4 , Gary S. Stein 4 , Rohini Muthuswami 2 , Anthony N. Imbalzano 1 , Jeffrey A. Nickerson 1 1 Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, MA, USA 2 School of Life Sciences, Jawaharlal Nehru University, New Delhi, Delhi, India 3 Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA 4 Department of Biochemistry and Vermont Cancer Center for Basic and Translational Research, University of Vermont College of Medicine, Burlington, WA, USA Correspondence to: Anthony N. Imbalzano, email: anthony.imbalzano@umassmed.edu Jeffrey A. Nickerson, email: jeffrey.nickerson@umassmed.edu Keywords: breast cancer, metabolism, lipogenesis, gene regulation, BRG1 Received: January 07, 2016 Accepted: May 01, 2016 Published: May 20, 2016 ABSTRACT Cancer cells reprogram cellular metabolism to meet the demands of growth. Identification of the regulatory machinery that regulates cancer-specific metabolic changes may open new avenues for anti-cancer therapeutics. The epigenetic regulator BRG1 is a catalytic ATPase for some mammalian SWI/SNF chromatin remodeling enzymes. BRG1 is a well-characterized tumor suppressor in some human cancers, but is frequently overexpressed without mutation in other cancers, including breast cancer. Here we demonstrate that BRG1 upregulates de novo lipogenesis and that this is crucial for cancer cell proliferation. Knockdown of BRG1 attenuates lipid synthesis by impairing the transcription of enzymes catalyzing fatty acid and lipid synthesis. Remarkably, exogenous addition of palmitate, the key intermediate in fatty acid synthesis, rescued the cancer cell proliferation defect caused by BRG1 knockdown. Our work suggests that targeting BRG1 to reduce lipid metabolism and, thereby, to reduce proliferation, has promise for epigenetic therapy in triple negative breast cancer.

This report represents the first study of keratinophilic fungi present in soils of Jamaica. Out of the 40 soil samples examined from different habitats, 30 (75%) were positive for the presence of keratinophilic fungi, yielding 36 isolates of keratinophilic fungi. Microsporum gypseum complex (represented by 16 isolates of M. gypseum, and four of M. fulvum) was most frequent, being present in 50% of the samples. A very high occurrence of this dermatophyte in Jamaican soil is of public health significance. The remaining isolates of keratinophilic fungi were represented by Chrysosporium spp (mainly C. indicum and C. tropicum) and Sepedonium sp. Esta comunicação representa o primeiro estudo sobre fungos queratinofílicos presentes em solos da Jamaica. De 40 amostras de solo examinadas de diferentes localidades, 30 (75%) foram positivas para a presença de fungos queratinofílicos permitindo 36 isolamentos dos mesmos. O complexo Microsporum gypseum (representados por 16 isolamentos de M. gypseum e quatro de M. fulvum) foi o mais frequente, estando presente em 50% das amostras. A muito alta ocorrência deste dermatófito no solo da Jamaica é significante para a saúde pública. Os isolados remanescentes de fungos queratinofílicos foram representados pelo Chrysosporium spp (principalmente C. indicum e C. tropicum) e Sepedonium sp.

Introduction: Information on the prevalence of keratinophilic fungi in West Indies is scanty. Occurrence of keratinophilic fungi in soils of St. Kitts and Nevis has not been investigated previously. Methodology: The prevalence of keratinophilic fungi was investigated in 108 samples of soils of varying habitats from St. Kitts and 55 such samples from Nevis by hair-baiting technique. Fungal growths appearing on the hair baits after four to eight weeks of incubation at room temperature were microscopically examined and cultured on mycological media. Cultures were identified on the basis of colonial and microscopic features. Results: Forty-nine (45%) of the samples from St. Kitts and 38 (69%) from Nevis were positive for keratinophilic fungi. Microsporum gypseum complex, a well-known geophilic dermatophyte, was the most frequently recovered species being present in 15.7% of soils of St. Kitts and 40% of soils of Nevis. The next commonest species recovered was Chrysosporium indicum , represented by 15 (13.9%) isolates from St. Kitts and seven (12.7%) isolates from Nevis. Other infrequently isolated keratinophilic species included Chrysosporium tropicum , Chrysosporium keratinophilum , and unidentified Chrysosporium species. Conclusions: This study is the first of its kind in the islands of St. Kitts and Nevis. A high incidence of M. gypseum complex in the soil of these islands is a noteworthy finding of public health significance.

Background Previously, we showed that aminoglycoside phosphotransferases catalyze the formation of a specific inhibitor of the SWI2/SNF2 proteins. Aminoglycoside phosphotransferases, for example neomycin-resistant genes, are used extensively as selection markers in mammalian transfections as well as in transgenic studies. However, introduction of the neomycin-resistant gene is fraught with variability in gene expression. We hypothesized that the introduction of neomycin-resistant genes into mammalian cells results in inactivation of SWI2/SNF2 proteins thereby leading to global epigenetic changes. Methodology Using fluorescence spectroscopy we have shown that the inhibitor, known as Active DNA-dependent ATPase A Domain inhibitor (ADAADi), binds to the SWI2/SNF2 proteins in the absence as well as presence of ATP and DNA. This binding occurs via a specific region known as Motif Ia leading to a conformational change in the SWI2/SNF2 proteins that precludes ATP hydrolysis. ADAADi is produced from a plethora of aminoglycosides including G418 and Streptomycin, two commonly used antibiotics in mammalian cell cultures. Mammalian cells are sensitive to ADAADi; however, cells stably transfected with neomycin-resistant genes are refractory to ADAADi. In resistant cells, endogenous SWI2/SNF2 proteins are inactivated which results in altered histone modifications. Microarray data shows that the changes in the epigenome are reflected in altered gene expression. The microarray data was validated using real-time PCR. Finally, we show that the epigenetic changes are quantized. Significance The use of neomycin-resistant genes revolutionized mammalian transfections even though questions linger about efficacy. In this study, we have demonstrated that selection of neomycin-resistant cells results in survival of only those cells that have undergone epigenetic changes, and therefore, data obtained using these resistant genes as selection markers need to be cautiously evaluated.

The dwindling fossil fuels have impressed upon the need of finding alternative energy sources. The biofuels derived from renewable resources are an interesting option which is being extensively researched. Biodiesel from non edible plant oils has emerged on the scene which can meet future energy demand. Jatropha curcas, a plant well adapted to semi arid condition is being seen as the source for biodiesel production. Many aspects of this plant have been studied right from its adaptation to genetic improvement. There are many reports where studies have been made on the variability at molecular level. This review deals with the biodiesel production from Jatropha at large and finally on the need of finding other plant sources that can serve the purpose of biodiesel production in the coming times.