Establishment and Characterization of a Highly Tumorigenic African American Prostate Cancer Cell Line,E006AA-hT

Genuine racial differences in prostate cancer (PCa) biology have been considered among the potential reasons to explain PCa disparities. There is no animal model to represent all aspects of human PCa and, more specifically, to be used for PCa disparity research. The lack of a spontaneously transformed in vitro cell-based model system has been a significant impediment to investigating and understanding potential molecular mechanisms, and the hormonal, genetic, and epigenetic factors underlying the biological and clinical aggressiveness of PCa in African American (AA) men.In this study, we established and characterized the E006AA-hT cell line as a highly tumorigenic subline of the previously characterized primary AA-PCa cell line, E006AA. Extensive characterization of the E006AA-hT cell line was accomplished using cytodifferentiation and prostate-specific markers, spectral karyotyping, cell line authentication assays, cell proliferation and migration assays, and in vitro tumorigenesis assays.Spectral karyotyping of E006AA-hT showed a hypertriploid chromosome complement and shared cytogenetic changes similar to its parental cells such as diploid X, absence of Y-chromosomes, numerical gains in chromosomes 5,6,8,10,17,20,21, and marker chromosomes of unknown origin. In addition, E006AA-hT also presented numerous clonal and structural aberrations such as insertion, deletion, duplication, and translocations in chromosomes 1-5, 8, 9, 11, 13, 14, 17, and 18. The E006AA-hT cell line was shown to be highly tumorigenic and produced tumors at an accelerated growth rate in both athymic nude and triple-deficient SCID mice.Silencing the mutated androgen receptor (AR-599 Ser>Gly) did not affect proliferation (loss-of-function), but decreased migration (gain-of-function) in E006AA-hT and its parental cell type. These data support that AR-point mutations may lead simultaneously to different “loss-of-function” and “gain-of-function” phenotypes in PCa cells. E006AA-Par and its subline as the only available spontaneously transformed low- and highly-tumorigenic primary AA-PCa cell lines could be used for basic and translational research aimed in supporting prostate cancer disparity research.     

Phosphorus availability increases pathobiome abundance and invasion of rhizosphere microbial networks by Ralstonia

Soil disease-suppressiveness depends on complex interactions among pathogens, native microbiota, and physicochemical properties, while these interactions remain understudied. Comparing field and microcosm experiments, we investigated the significance of these interactions in disease emergence or suppression using structural equation modelling (SEM) and receiver operating characteristic curve (ROC) analyses. We observed significant differences in the relative abundance of pathogenic and beneficial microbes, alpha and beta diversity indices between disease-conducive and -suppressive rhizosphere soils. The pathogenic (Ralstonia) and beneficial (Bacillus) taxa dominated disease-conducive and -suppressive rhizosphere soils, respectively. Moreover, the co-occurrences of Ralstonia with native microorganisms were positive and negative in the disease-conducive and -suppressive soils, respectively. These results suggest the supportive (Rudaea) and suppressive (Enterobacter, Bacillus) role of indigenous microbes in the invasion of soil and plant systems by Ralstonia. The SEM and ROC analysis predicted that Ralstonia invaded rhizospheric microbial networks and caused peanut wilt under high than low soil phosphorus conditions. Our results suggest the importance of soil phosphorus availability in altering the microbial interactions, thus leading to soil invasion by Ralstonia. Thus, we conclude by saying that feeding soil with high amounts of available phosphorus could deplete plant-beneficial microbes and increase the pathobiome abundance that may compromise plant health.     

Force and twist dependence of RepC nicking activity on torsionally-constrained DNA molecules

Many bacterial plasmids replicate by an asymmetric rolling-circle mechanism that requires sequence-specific recognition for initiation, nicking of one of the template DNA strands and unwinding of the duplex prior to subsequent leading strand DNA synthesis. Nicking is performed by a replication-initiation protein (Rep) that directly binds to the plasmid double-stranded origin and remains covalently bound to its substrate 5′-end via a phosphotyrosine linkage. It has been proposed that the inverted DNA sequences at the nick site form a cruciform structure that facilitates DNA cleavage. However, the role of Rep proteins in the formation of this cruciform and the implication for its nicking and religation functions is unclear. Here, we have used magnetic tweezers to directly measure the DNA nicking and religation activities of RepC, the replication initiator protein of plasmid pT181, in plasmid sized and torsionally-constrained linear DNA molecules. Nicking by RepC occurred only in negatively supercoiled DNA and was force- and twist-dependent. Comparison with a type IB topoisomerase in similar experiments highlighted a relatively inefficient religation activity of RepC. Based on the structural modeling of RepC and on our experimental evidence, we propose a model where RepC nicking activity is passive and dependent upon the supercoiling degree of the DNA substrate.     

Punica granatum waste to ethanol valorisation employing optimized levels of saccharification and fermentation

Pomegranate peels (PPW) as municipal waste is inexpensive biomass that could be a renewable source of sugars particularly rich in hemicellulosic contents. The subsequent conversion of available sugars in PPW can provide prospective strategy for cost-effective bioenergy production. In this study, an experimental setup based on CCD was implemented with the aim of bioconversion of biomass into bioethanol. The factors considered were Hydrochloric acid concentration (X₁), the hydrolysis temperature (X₂) and time (X₃) for optimization with dilute Hydrochloric acid (HCl) saccharification. The present study investigates the optimised level of bioethanol synthesis from acid pre-treated PPW explained by RSM. Subsequently, three yeasts viz. Saccharomyces cerevisiae K7, Metschnikowia sp. Y31 and M. cibodasensis Y34 were utilized for fermentation of acid hydrolysed and detoxified feed stocks. Optimum values of reducing sugars 48.02 ± 0.02 (gL^?1) and total carbohydrates 205.88 ± 0.13 (gL^?1) were found when PPW was hydrolyzed with 1% HCl concentration at 100?C of temperature for 30 min. Later on, fermentation of PPWH after detoxification with 2.5% activated charcoal. The significant ethanol (g ethanol/g of reducing sugars) yields after fermentation with Metschnikowia sp. Y31 and M. cibodasensis Y34 found to be 0.40 ± 0.03 on day 5 and 0.41 ± 0.02 on last day of experiment correspondingly. Saccharomyces cerevisiae K7 also produce maximum ethanol 0.40 ± 0.00 on last day of incubation utilizing the PPWH. The bioconversion of commonly available PPW into bioethanol as emphasize in this study could be a hopeful expectation and also cost-effective to meet today energy crisis.     

Interactive role of zinc and iron lysine on Spinacia oleracea L. growth,photosynthesis and antioxidant capacity irrigated with tannery wastewater

AbstractUntreated wastewater contains toxic amounts of heavy metals such as chromium (Cr), which poses a serious threat to the growth and physiology of plants when used in irrigation. Though, Cr is among the most widespread toxic trace elements found in agricultural soils due to various anthropogenic activities. To explore the interactive effects of micronutrients with amino acid chelators [iron-lysine (Fe-lys) and zinc-lysine (Zn-lys)], pot experiments were conducted in a controlled environment, using spinach (Spinacia oleracea L.) plant irrigated with tannery wastewater. S. oleracea was treated without Fe and Zn-lys (0 mg/L Zn-lys and 0 mg/L Fe-lys) and also treated with various combinations of (interactive application) Fe and Zn-lys (10 mg/L Zn-lys and 5 mg/L Fe-lys), when cultivated at different levels [0 (control) 33, 66 and 100%) of tannery wastewater in the soil having a toxic level of Cr in it. According to the results, we have found that, high concentration of Cr in the soil significantly (P < 0.05) reduced plant height, fresh biomass of roots and leaves, dry biomass of roots and leaves, root length, number of leaves, leaf area, total chlorophyll contents, carotenoid contents, transpiration rate (E), stomatal conductance (gs), net photosynthesis (PN), and water use efficiency (WUE) and the contents of Zn and Fe in the plant organs without foliar application of Zn and Fe-lys. Moreover, phytotoxicity of Cr increased malondialdehyde (MDA) contents in the plant organs (roots and leaves), which induced oxidative damage in S. oleracea manifested by the contents of hydrogen peroxide (H₂O₂) and membrane leakage. The negative effects of Cr toxicity could be overturned by Zn and Fe-lys application, which significantly (P < 0.05) increase plant growth, biomass, chlorophyll content, and gaseous exchange attributes by reducing oxidative stress (H₂O₂, MDA, EL) and increasing the activities of various antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD) catalase (CAT) and ascorbate peroxidase (APX). Furthermore, the supplementation of Zn and Fe-lys increased the contents of essential nutrients (Fe and Zn) and decreased the content of Cr in all plant parts compared to the plants cultivated in tannery wastewater without application of Fe-lys. Taken together, foliar supplementation of Zn and Fe-lys alleviates Cr toxicity in S. oleracea by increased morpho-physiological attributes of the plants, decreased Cr contents and increased micronutrients uptake by the soil, and can be an effective in heavy metal toxicity remedial approach for other crops.Graphic abstract     

Isolation and analysis of the total genomic DNA from the date palm (P. dactylifera L.) and related species

The objective of this study was to adapt a plant DNA preparation procedure for the isolation of biologically active DNA and DNA with a high molecular weight from the date palm and other related palms. Mature leaf tissue extractions of the date palm, Phoenix dactylifera L., the coconut tree, Cocos nucifera, and the Mexican Fan Palm, Washingtonia robusta, were characterized for total genomic DNA yield, purity, integrity, as well as restriction digestion and ligation capabilities. It is demonstrated here that the DNA isolation procedure, modified for use with various palm leaf tissues, met the criteria for simplicity and low costs, and yielded DNA of high molecular weight (～50 Kbp) and of sufficient purity suitable for molecular studies.