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Brain cancer is regarded as one of the most life-threatening forms of cancer worldwide. Oxidative stress acts to derange normal brain homeostasis, thus is involved in carcinogenesis in brain. The Nrf2/Keap1/ARE pathway is an important signaling cascade responsible for the maintenance of redox homeostasis, and regulation of anti-inflammatory and anticancer activities by multiple downstream pathways. Interestingly, Nrf2 plays a somewhat, contradictory role in cancers, including brain cancer. Nrf2 has traditionally been regarded as a tumor suppressor since its cytoprotective functions are considered to be the principle cellular defense mechanism against exogenous and endogenous insults, such as xenobiotics and oxidative stress. However, hyperactivation of the Nrf2 pathway supports the survival of normal as well as malignant cells, protecting them against oxidative stress, and therapeutic agents. Plants possess a pool of secondary metabolites with potential chemotherapeutic/chemopreventive actions. Modulation of Nrf2/ARE and downstream activities in a Keap1-dependant manner, with the aid of plant-derived secondary metabolites exhibits promise in the management of brain tumors. Current article highlights the effects of Nrf2/Keap1/ARE cascade on brain tumors, and the potential role of secondary metabolites regarding the management of the same. [ABSTRACT FROM AUTHOR]

Abstract Brain cancer is regarded as one of the most life-threatening forms of cancer worldwide. Oxidative stress acts to derange normal brain homeostasis, thus is involved in carcinogenesis in brain. The Nrf2/Keap1/ARE pathway is an important signaling cascade responsible for the maintenance of redox homeostasis, and regulation of anti-inflammatory and anticancer activities by multiple downstream pathways. Interestingly, Nrf2 plays a somewhat, contradictory role in cancers, including brain cancer. Nrf2 has traditionally been regarded as a tumor suppressor since its cytoprotective functions are considered to be the principle cellular defense mechanism against exogenous and endogenous insults, such as xenobiotics and oxidative stress. However, hyperactivation of the Nrf2 pathway supports the survival of normal as well as malignant cells, protecting them against oxidative stress, and therapeutic agents. Plants possess a pool of secondary metabolites with potential chemotherapeutic/chemopreventive actions. Modulation of Nrf2/ARE and downstream activities in a Keap1-dependant manner, with the aid of plant-derived secondary metabolites exhibits promise in the management of brain tumors. Current article highlights the effects of Nrf2/Keap1/ARE cascade on brain tumors, and the potential role of secondary metabolites regarding the management of the same.

Abstract Heat-shock protein beta1 (HSPB1) is a member of the small HSP family, downregulated in osteoarthritis (OA) chondrocytes and demonstrated the capacity to serve as an RNA-binding protein (RBP). This work aimed to explore the profile of HSPB1 bound RNA and reveal the potential regulation mechanism of HSPB1 in OA. In this work, we captured an unbiased HSPB1-RNA interaction map in Hela cells using the iRIP-seq. The results demonstrated that HSPB1 interacted with plentiful of mRNAs and genomic location toward the CDS region. Functional enrichment of HSPB1-related peaks showed the involvement in gene expression, translation initiation, cellular protein metabolic process, and nonsense-mediated decay. HOMER software analysis showed that HSPB1 bound peaks were over-represented in GAGGAG sequences. In addition, ABLIRC and CIMS algorithm indicated that HSPB1 bound to AU-rich motifs and the proportion of AU-rich peaks in 3′ UTR were slightly higher than that in other regions. Moreover, HSPB1-binding targets analysis revealed several gens were associated with OA including EGFR, PLEC, COL5A1, and ROR2. The association of OA-related mRNAs to HSPB1 was additionally confirmed in OA tissues by the quantitative RIP-PCR experiments. Further experiment demonstrated the downregulation of HSPB1 in OA tissues. In conclusion, our current study confirmed HSPB1 as an RNA-binding protein and revealed its potential function in the pathological process of OA, providing a reliable insight to further investigate the molecular regulation mechanism of HSPB1 in OA.

Heat-shock protein beta1 (HSPB1) is a member of the small HSP family, downregulated in osteoarthritis (OA) chondrocytes and demonstrated the capacity to serve as an RNA-binding protein (RBP). This work aimed to explore the profile of HSPB1 bound RNA and reveal the potential regulation mechanism of HSPB1 in OA. In this work, we captured an unbiased HSPB1-RNA interaction map in Hela cells using the iRIP-seq. The results demonstrated that HSPB1 interacted with plentiful of mRNAs and genomic location toward the CDS region. Functional enrichment of HSPB1-related peaks showed the involvement in gene expression, translation initiation, cellular protein metabolic process, and nonsense-mediated decay. HOMER software analysis showed that HSPB1 bound peaks were over-represented in GAGGAG sequences. In addition, ABLIRC and CIMS algorithm indicated that HSPB1 bound to AU-rich motifs and the proportion of AU-rich peaks in 3′ UTR were slightly higher than that in other regions. Moreover, HSPB1-binding targets analysis revealed several gens were associated with OA including EGFR, PLEC, COL5A1, and ROR2. The association of OA-related mRNAs to HSPB1 was additionally confirmed in OA tissues by the quantitative RIP-PCR experiments. Further experiment demonstrated the downregulation of HSPB1 in OA tissues. In conclusion, our current study confirmed HSPB1 as an RNA-binding protein and revealed its potential function in the pathological process of OA, providing a reliable insight to further investigate the molecular regulation mechanism of HSPB1 in OA. [ABSTRACT FROM AUTHOR]

Metabolic and epigenetic reprogramming play important roles in cancer therapeutic resistance. However, their interplays are poorly understood. We report here that elevated TIGAR (TP53-induced glycolysis and apoptosis regulator), an antioxidant and glucose metabolic regulator and a target of oncogenic histone methyltransferase NSD2 (nuclear receptor binding SET domain protein 2), is mainly localized in the nucleus of therapeutic resistant tumor cells where it stimulates NSD2 expression and elevates global H3K36me2 mark. Mechanistically, TIGAR directly interacts with the antioxidant master regulator NRF2 and facilitates chromatin recruitment of NRF2, H3K4me3 methylase MLL1 and elongating Pol-II to stimulate the expression of both new (NSD2) and established (NQO1/2, PRDX1 and GSTM4) targets of NRF2, independent of its enzymatic activity. Nuclear TIGAR confers cancer cell resistance to chemotherapy and hormonal therapy in vitro and in tumors through effective maintenance of redox homeostasis. In addition, nuclear accumulation of TIGAR is positively associated with NSD2 expression in clinical tumors and strongly correlated with poor survival. These findings define a nuclear TIGAR-mediated epigenetic autoregulatory loop in redox rebalance for tumor therapeutic resistance.

Sedanolide is a bioactive compound with anti-inflammatory and antitumor activities. Although it has been recently suggested that sedanolide activates the nuclear factor E2-related factor 2 (NRF2) pathway, there is little research on its effects on cellular resistance to oxidative stress. The objective of the present study was to investigate the function of sedanolide in suppressing hydrogen peroxide (H2O2)-induced oxidative damage and the underlying molecular mechanisms in human hepatoblastoma cell line HepG2 cells. We found that sedanolide activated the antioxidant response element (ARE)-dependent transcription mediated by the nuclear translocation of NRF2. Pathway enrichment analysis of RNA sequencing data revealed that sedanolide upregulated the transcription of antioxidant enzymes involved in the NRF2 pathway and glutathione metabolism. Then, we further investigated whether sedanolide exerts cytoprotective effects against H2O2-induced cell death. We showed that sedanolide significantly attenuated cytosolic and mitochondrial reactive oxygen species (ROS) generation induced by exposure to H2O2. Furthermore, we demonstrated that pretreatment with sedanolide conferred a significant cytoprotective effect against H2O2-induced cell death probably due to preventing the decrease in the mitochondrial membrane potential and the increase in caspase-3/7 activity. Our study demonstrated that sedanolide enhanced cellular resistance to oxidative damage via the activation of the Kelch-like ECH-associated protein 1 (KEAP1)–NRF2 pathway. [ABSTRACT FROM AUTHOR]

Flavone is widely found in plants and plays an important role in plant defense against pests. Many pests, such as Helicoverpa armigera, use flavone as a cue to upregulate counter-defense genes for detoxification of flavone. Yet the spectrum of the flavone-inducible genes and their linked cis-regulatory elements remains unclear. In this study, 48 differentially expressed genes (DEGs) were found by RNA-seq. These DEGs were mainly concentrated in the retinol metabolism and drug metabolism-cytochrome P450 pathways. Further in silico analysis of the promoter regions of 24 upregulated genes predicted two motifs through MEME and five previously characterized cis-elements including CRE, TRE, EcRE, XRE-AhR and ARE. Functional analysis of the two predicted motifs and two different versions of ARE (named ARE1 and ARE2) in the promoter region of the flavone-inducible carboxylesterase gene CCE001j verified that the two motifs and ARE2 are not responsible for flavone induction of H. armigera counter-defense genes, whereas ARE1 is a new xenobiotic response element to flavone (XRE-Fla) and plays a decisive role in flavone induction of CCE001j. This study is of great significance for further understanding the antagonistic interaction between plants and herbivorous insects. [ABSTRACT FROM AUTHOR]

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For testing goodness of fit, we consider a class of U-statistics of overlapping spacings of order two, and investigate their asymptotic properties. The standard U-statistic theory is not directly applicable here as the overlapping spacings form a dependent random sequence. The asymptotic distribution of the statistics under the null hypothesis and under a sequence of local alternatives are derived. In terms of the Pitman ARE, the U-statistic based on Gini's mean square difference of overlapping spacings is found to be the asymptotically locally most powerful. Interestingly, this test has the same efficacy as the Greenwood test based on overlapping spacings. • For testing goodness of fit, we consider U-statistics of overlapping spacings. • We derive asymptotic distribution of statistics for a sequence of local alternatives. • The asymptotically locally most powerful test is obtained in terms of the Pitman ARE. [ABSTRACT FROM AUTHOR]

Living organisms have developed finely regulated homeostatic networks to mitigate the effects of environmental fluctuations in transition metal micronutrients, including iron, zinc, and copper. In Saccharomyces cerevisiae , the tandem zinc-finger protein Cth2 post-transcriptionally regulates gene expression under conditions of iron deficiency by controlling the levels of mRNAs that code for non-essential ferroproteins. The molecular mechanism involves Cth2 binding to AU-rich elements present in the 3′ untranslated region of target mRNAs, negatively affecting their stability and translation. Arabidopsis thaliana has two TZF proteins homologous to yeast Cth2, C3H14 and C3H15, which participate in cell wall remodelling. The present work examines the expression of representative metal homeostasis genes with putative AREs in plants with altered levels of C3H14 and C3H15 grown under varying metal availabilities. The results suggest that C3H15 may act as a post-transcriptional plant modulator of metal adequacy, as evidenced by the expression of SPL7 , the main transcriptional regulator under copper deficiency, and PETE2 , which encodes plastocyanin. In contrast to S. cerevisiae , the plant C3H15 affects copper and zinc homeostasis rather than iron. When grown under copper-deficient conditions, adult C3H15 OE plants exhibit lower chlorophyll content and photosynthetic efficiency compared to control plants, suggesting accelerated senescence. Likewise, metal content in C3H15 OE plants under copper deficiency shows altered mobilization of copper and zinc to seeds. These data suggest that the C3H15 protein plays a role in modulating both cell wall remodelling and metal homeostasis. The interaction between these processes may be the cause of altered metal translocation. • C3H15 functions as a post-transcriptional regulator in Arabidopsis copper and zinc homeostasis. • A comparison of the TZF domains of S. cerevisiae Cth2 and higher plants reveals that the TZF1 is most conserved than TZF2. • Analysis of Arabidopsis genes with putative C3H15 target sequences reveals that metal homeostasis genes are overrepresented. • Under copper deficiency, C3H15 OE show an accelerated senescence as indicated by chlorophyll content and fluorescence. • Seeds of C3H15OE contain reduced zinc and copper levels pointing to their difficulty in mobilizing copper. [ABSTRACT FROM AUTHOR]

Sedanolide is a bioactive compound with anti-inflammatory and antitumor activities. Although it has been recently suggested that sedanolide activates the nuclear factor E2-related factor 2 (NRF2) pathway, there is little research on its effects on cellular resistance to oxidative stress. The objective of the present study was to investigate the function of sedanolide in suppressing hydrogen peroxide (H2O2)-induced oxidative damage and the underlying molecular mechanisms in human hepatoblastoma cell line HepG2 cells. We found that sedanolide activated the antioxidant response element (ARE)-dependent transcription mediated by the nuclear translocation of NRF2. Pathway enrichment analysis of RNA sequencing data revealed that sedanolide upregulated the transcription of antioxidant enzymes involved in the NRF2 pathway and glutathione metabolism. Then, we further investigated whether sedanolide exerts cytoprotective effects against H2O2-induced cell death. We showed that sedanolide significantly attenuated cytosolic and mitochondrial reactive oxygen species (ROS) generation induced by exposure to H2O2. Furthermore, we demonstrated that pretreatment with sedanolide conferred a significant cytoprotective effect against H2O2-induced cell death probably due to preventing the decrease in the mitochondrial membrane potential and the increase in caspase-3/7 activity. Our study demonstrated that sedanolide enhanced cellular resistance to oxidative damage via the activation of the Kelch-like ECH-associated protein 1 (KEAP1)–NRF2 pathway.

The detoxification of quinones through NAD(P)H:quinone oxidoreductase (NQO1) is a crucial mechanism to maintain cellular homeostasis. The exposure to heavy metals, specifically methylmercury (MeHg), induces several antioxidant enzymes, including NQO1. The nuclear factor erythroid 2-related factor-2 (NRF2) is known to regulate the expression of Nqo1 gene and also the aryl hydrocarbon receptor (AHR) is another Nqo1 gene regulator. This co-regulation prompted us to investigate which transcription factor (NRF2 or AHR) orchestrates the regulation of NQO1 expression upon MeHg exposure. Therefore, we investigated how MeHg can modulate the level of NQO1 expression by exposing Hepa-1c1c7 cells to several concentrations of MeHg with and without the addition of NQO1 inducers, DL-sulforaphane (SUL) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). We found that the mRNA expression of Nqo1 is up-regulated by MeHg in time- as well as dose-dependent fashions. Additionally, MeHg increased the NQO1 at all expression levels with and without the presence of its inducers, SUL or TCDD. Furthermore, the MeHg-mediated increase of NQO1 expression was in parallel with a concurrent increase in the nuclear localization of NRF2 protein, but not that of AHR. Mechanistically, the antioxidant response element-driven reporter gene activity was induced by 215% upon MeHg exposure. Also, transfecting Hepa-1c1c7 with Nrf2 siRNA reduced the MeHg-induced NQO1 protein expression by 60%. In conclusion, our findings provide evidence supporting the hypothesis that MeHg upregulates the Nqo1 gene through a transcriptional mechanism at least in part via a NRF2-dependent mechanism.

Insecticides have potential to non-target organisms, disrupting the healthy functioning of the aquatic environment as they are the ultimate receptor of the aquatic ecosystem. Insecticides, which are widely used in agriculture, have high neurotoxicity on aquatic organisms. In this study, the acute alterations [catalase (CAT), arylesterase (ARE), malondialdehyde (MDA), myeleperoxidase (MPO), paraoxonase (PON), glutathione peroxidase (GPX), superoxide dismutase (SOD), 8-hydroxy-2-deoxyguanosine (8-OHdG) level, caspase-3 activity, and Acetylcholinesterase (AChE) enzyme activity] caused by the different concentrations of Fipronil (FP) insecticide (0.05, 0.1, and 0.2 mg/L) on rainbow trout (Oncorhynchus mykiss) brain tissue were investigated. It has been determined that superoxide dismutase –catalase - glutathione peroxidase - paraoxonase and arylesterase enzyme activities were inhibited but MDA and MPO induced depending on the concentration in brain tissue. When compared with the control group, the changes between the pesticide exposed groups were found statistically significant (p < 0.05). In brain tissue, while AChE enzyme activity was decreased depending on concentration, caspase-3 activity increased with 8-OHdG level. As a result, it has been determined that FP is a dangerous environmental pollutant for aquatic organisms, even at low concentrations, inducing oxidative stress, damaging the brain tissue of fish and stimulating apoptosis. [ABSTRACT FROM AUTHOR]

Abstract Background Severe sepsis, a major health problem worldwide, has become one of the leading causes of death in ICU patients. Further study on the pathogenesis and treatment of acute kidney injury (AKI) is of great significance to reduce high mortality rate of sepsis. In this study, the mechanism by which ubiquitin specific peptidase 10 (USP10) reduces sepsis-induced AKI was investigated. Ligation and perforation of cecum (CLP) was employed to establish C57BL/6 mouse models of sepsis. Hematoxylin-eosin (H&E) staining was performed to detect renal injury. The concentrations of serum creatinine (Cr), urea nitrogen (BUN) and cystatin C (Cys C) were determined using a QuantiChrom™ Urea Assay kit. RT-qPCR and western blot were conducted to assess the USP10 expression level. DHE staining was used to detect reactive oxygen species (ROS) levels. H2O2, MDA and SOD levels were assessed using corresponding colorimetric kits. Western blot was used to examine the expression levels of Bcl-2, Bax, cleaved caspase-3, Sirt6, Nrf2 and HO-1. MTT assay was used to determine cell viability, whereas TUNEL staining and flow cytometry were used to assess cell apoptosis. Results In this study, we found that USP10 was decreased in CLP-induced mouse renal tissues. We identified that USP10 alleviated renal dysfunction induced by CLP. Moreover, USP10 was found to reduce oxidative stress, and abated LPS-induced renal tubular epithelial cell injury and apoptosis. Finally, we discovered that USP10 promoted activation of the NRF2/HO-1 pathway through SIRT6 and attenuated LPS-induced renal tubular epithelial cell injury. Conclusions This study found that USP10 activates the NRF2/ARE signaling through SIRT6. USP10 alleviates sepsis-induced renal dysfunction and reduces renal tubular epithelial cell apoptosis and oxidative stress.

The KEAP1-NRF2-ARE signaling pathway plays a central role in mediating the adaptive cellular stress response to oxidative and electrophilic chemicals. This canonical pathway has been extensively studied and reviewed in the past two decades, but rarely was it looked at from a quantitative signaling perspective. Signal amplification, i.e., ultrasensitivity, is crucially important for robust induction of antioxidant genes to appropriate levels that can adequately counteract the stresses. In this review article, we examined a number of well-known molecular events in the KEAP1-NRF2-ARE pathway from a quantitative perspective with a focus on how signal amplification can be achieved. We illustrated, by using a series of mathematical models, that redox-regulated protein sequestration, stabilization, translation, nuclear trafficking, DNA promoter binding, and transcriptional induction – which are embedded in the molecular network comprising KEAP1, NRF2, sMaf, p62, and BACH1 – may generate highly ultrasensitive NRF2 activation and antioxidant gene induction. The emergence and degree of ultrasensitivity depend on the strengths of protein-protein and protein-DNA interaction and protein abundances. A unique, quantitative understanding of signal amplification in the KEAP1-NRF2-ARE pathway will help to identify sensitive targets for the prevention and therapeutics of oxidative stress-related diseases and develop quantitative adverse outcome pathway models to facilitate the health risk assessment of oxidative chemicals.

Flavone is widely found in plants and plays an important role in plant defense against pests. Many pests, such as Helicoverpa armigera, use flavone as a cue to upregulate counter-defense genes for detoxification of flavone. Yet the spectrum of the flavone-inducible genes and their linked cis-regulatory elements remains unclear. In this study, 48 differentially expressed genes (DEGs) were found by RNA-seq. These DEGs were mainly concentrated in the retinol metabolism and drug metabolism-cytochrome P450 pathways. Further in silico analysis of the promoter regions of 24 upregulated genes predicted two motifs through MEME and five previously characterized cis-elements including CRE, TRE, EcRE, XRE-AhR and ARE. Functional analysis of the two predicted motifs and two different versions of ARE (named ARE1 and ARE2) in the promoter region of the flavone-inducible carboxylesterase gene CCE001j verified that the two motifs and ARE2 are not responsible for flavone induction of H. armigera counter-defense genes, whereas ARE1 is a new xenobiotic response element to flavone (XRE-Fla) and plays a decisive role in flavone induction of CCE001j. This study is of great significance for further understanding the antagonistic interaction between plants and herbivorous insects.

BACKGROUND: Tissue damage induced by radioactive ionizing radiation is one of the serious complications of radiotherapy. Activation of endogenous antioxidative stress response through nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/antioxidant responsive element (ARE) signaling pathway has shown preventive and therapeutic effects on radiation-induced tissue injuries. OBJECTIVE: To explain the preventive and therapeutic mechanisms of Nrf2/ARE signaling pathway on radiation-induced injuries, and provide new ideas for exploring the treatment and prevention strategies of radiotherapy complications. METHODS: PubMed, Web of Science, CNKI, WanFang and VIP databases were searched using the keywords of “radiation injury, radiation-induced tissue injury (intestine, liver, tongue, lung, bone marrow, skin, hematopoietic system, nervous system, circulatory system), radiation damage, radiation syndrome, Nrf2” in Chinese and English, respectively. A total of 67 literatures were finally included and reviewed. RESULTS AND CONCLUSION: Radiation-induced injury is damage to tissues caused by the intramolecular bond homolysis of ionizing radiation biomolecules that lead to an increase in cytotoxic free radicals and reactive oxygen species. TheNrf2/ARE signaling pathway is an important endogenous antioxidant defense mechanism in the body. By enhancing the expression of antioxidant response elements, it can reduce DNA damage, promote damaged DNA repair, and inhibit radiation-induced cell apoptosis. Therefore, the Nrf2/ARE signaling pathway is of great significance for the prevention and correction of cellular redox imbalances including radioactive tissue damage. Current studies have found that the activation of Nrf2/ARE signaling pathway has two sides. On the one hand, the application of Nrf2/ARE signaling pathway activator cannot only prevent tissue loss caused by radiotherapy, but also induce the repair of damaged tissue after radiotherapy. On the other hand, the long-term and chronic activation of the Nrf2/ARE signaling pathway caused by various reasons may reduce the sensitivity of malignant tumors to radiotherapy and chemotherapy. Therefore, the Nrf2 /ARE signaling pathway has potential as a new strategy for the prevention and treatment of radiotherapy complications; however, whether it can be applied in clinical practice still needs further exploration and research. [ABSTRACT FROM AUTHOR]

One of the primary mechanisms of post-transcriptional gene regulation is the modulation of RNA stability. We recently discovered that LINC00675, a transcript annotated as a long noncoding RNA (lncRNA), is transcriptionally regulated by FOXA1 and encodes a highly conserved small protein that localizes to the endoplasmic reticulum, hence renamed as FORCP (FOXA1-regulated conserved small protein). Here, we show that the endogenous FORCP transcript is rapidly degraded and rendered unstable as a result of 3'UTR-mediated degradation. Surprisingly, although the FORCP transcript is a canonical nonsense-mediated decay (NMD) and microRNA (miRNA) target, we found that it is not degraded by NMD or miRNAs. Targeted deletion of an evolutionarily conserved region in the FORCP 3'UTR using CRISPR/Cas9 significantly increased the stability of the FORCP transcript. Interestingly, this region requires the presence of an immediate downstream 55-nt-long sequence for transcript stability regulation. Functionally, colorectal cancer cells lacking this conserved region expressed from the endogenous FORCP locus displayed decreased proliferation and clonogenicity. These data demonstrate that the FORCP transcript is destabilized via conserved elements within its 3'UTR and emphasize the need to interrogate the function of a given 3'UTR in its native context. [ABSTRACT FROM AUTHOR]

Nuclear factor erythroid 2-related factor-2 (Nrf2), and the less well characterised proteins Nrf1 and Nrf3, are member of the cap 'n' collar family of transcription factors. Nrf proteins regulate the expression of endogenous antioxidant enzymes and have recently become the targets for various therapeutic treatments. Recently, Nrf proteins have been of particular interest as a target in placental-derived oxidative stress induced pregnancy disorders. Here, we report the presence of Nrf1, Nrf2 and Nrf3 proteins in both human primary trophoblast and human trophoblast choriocarcinoma cell line (BeWo). We also detail the steps taken to successfully silence all Nrf proteins in both human primary trophoblast cells and BeWo via detection of mRNA and protein using quantitative PCR, and SDS-PAGE and Western Blotting respectively. [ABSTRACT FROM AUTHOR]

Concordant transcriptional regulation can generate multiple gene products that collaborate to achieve a common goal. Here we report a case of concordant transcriptional regulation that instead drives a single protein to be produced in the same cell type from divergent promoters. This gene product—the RHOX5 homeobox transcription factor—is translated from 2 different mRNAs with different 5′ untranslated regions (UTRs) transcribed from alternative promoters. Despite the fact that these 2 promoters—the proximal promoter (Pp) and the distal promoter (Pd)—exhibit different patterns of tissue-specific activity, share no obvious sequence identity, and depend on distinct transcription factors for expression, they exhibit a remarkably similar expression pattern in the testes. In particular, both depend on androgen signaling for expression in the testes, where they are specifically expressed in Sertoli cells and have a similar stage-specific expression pattern during the seminiferous epithelial cycle. We report evidence for 3 mechanisms that collaborate to drive concordant Pp / Pd expression. First, both promoters have an intrinsic ability to respond to androgen receptor and androgen. Second, the Pp acts as an enhancer to promote androgen-dependent transcription from the Pd. Third, Pd transcription is positively autoregulated by the RHOX5 protein, which is first produced developmentally from the Pp. Together, our data support a model in which the Rhox5 homeobox gene evolved multiple mechanisms to activate both of its promoters in Sertoli cells to produce Rhox5 in an androgen-dependent manner during different phases of spermatogenesis. [ABSTRACT FROM AUTHOR]

The oxidative polymerizations of 2-methyl-m-phenylenediamine were performed to synthesize PMPDIA-A and PMPDIA-B polymers by H2O2 (35% aqueous solution) oxidant in acid-catalyzed ethanol and basic aqueous medium. Furthermore, PMPDIA-E polymer of 2-methyl-m-phenylenediamine was enzymatically synthesized by HRP (horseradish peroxidase) enzyme and H2O2 (35% aqueous solution) in dioxane/0.1 M phosphate tampon solution (pH = 7) mixture. The structures of 2-methyl-m-phenylenediamine and polymers were confirmed by FT-IR, NMR and UV–Vis spectrometer measurements. Molecular weight distributions, surface morphologies, thermal and fluorescence properties of polymers were determined from GPC, SEM, TG-DTA, DSC and fluorescence spectra analyses, respectively. CV and UV–Vis analyses were performed to determine the HOMO–LUMO energy levels, electrochemical (Eg′) and optical (Eg) band gaps values of MPDIA, PMPDIA-E, PMPDIA-A and PMPDIA-B. The electrochemical and optical band gaps values of polymers were lower than MPDIA, because of their polyconjugated structures. According to GPC measurements of polymers, the weight average molecular weights were between 5400 and 10400 Da. The fluorescence quantum yield of PMPDIA-E was calculated to be 9.15% in DMSO solution. The adsorption of methylene blue (MB) from aqueous solution on PMPDIA-B was studied. Adsorption isotherms and equilibrium adsorption capacities were determined by the fitting of the experimental data to two well-known isotherm models: Langmuir and Freundlich.

NAD(P)H:quinone oxidoreductase 1 (NQO1) is considered as one of the most significant enzyme in cellular defense due to its ability to detoxify reactive quinones and quinone imines to their less toxic hydroquinones forms. NQO1 is a xenobiotic metabolizing cytosolic enzyme that catalyzes the reduction of two- or four-electron of numerous exogenous and endogenous quinones by utilizing flavin adenine dinucleotide (FAD) as a cofactor. NQO1 enzyme exists as a homodimer enzyme and is biochemically identified by its noticeable ability to utilize either NADH or NADPH as reducing cofactors and by its inhibition by anticoagulant agents such as dicumarol. NQO1 is a distinctly inducible enzyme and known to be controlled by the Nrf2-Keap1 pathway. The importance of the antioxidant activities exhibited by NQO1 enzyme in suppressing the oxidative stress status is provided by demonstration that induction or reduction of NQO1 levels are linked with increased and reduced susceptibilities to oxidative stress, respectively. The gene coding for NQO1 has two well-recognized polymorphisms at nucleotide site 609(C-T) and 465 (C-T) of the human cDNA. C609T causes complete loss of enzymatic activity due to protein instability, whereas the C465T results in reduction of the enzyme activity. In this Review, we deliberate the protecting activates of NQO1 and discussing its possible transcriptional pathways regulating its induction by Nrf2-Keap1/ARE system.

Background Age-related macular degeneration (AMD) is the leading cause of blindness in elderly population in the developed world. Dysfunction of retinal pigment epithelium (RPE) likely triggers early AMD stages. The effect of Quercetin on the early AMD in-vitro model remained unclear.Methods The effect of Quercetin in the cell viability was detected with CCK8 methods in control, CSE treated, and CSE with Quercetin treatment group. The apoptotic status in each group was detected with tunnel assay. The oxidative and inflammation biomarkers were detected by ELISA. The expression levels of Keap1/Nrf2/ARE in RPE cells were measured by western blot after pretreatment of Quercetin followed by CSE treatment.Results It was found that Quercetin could improve the cell viability and decrease cellular apoptotic rate in the CSE treated RPE group. The expressions of inflammatory and apoptotic biomarkers were significantly decreased in Quercetin treatment group. Furthermore, Quercetin exerts protective effects via activation Keap1/Nrf2/ARE pathway in CSE treated RPE cells.Conclusions Quercetin demonstrated significant protective effects in an in-vitro model of early AMD and it might be a new therapeutic strategy for the management of early AMD.

ROS, RNS, and carcinogenic metabolites generate excessive oxidative stress, which changes the basal cellular status and leads to epigenetic modification, genomic instability, and initiation of cancer. Epigenetic modification may inhibit tumor-suppressor genes and activate oncogenes, enabling cells to have cancer promoting properties. The nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that in humans is encoded by the NFE2L2 gene, and is activated in response to cellular stress. It can regulate redox homoeostasis by expressing several cytoprotective enzymes, including NADPH quinine oxidoreductase, heme oxygenase-1, UDP-glucuronosyltransferase, glutathione peroxidase, glutathione-S-transferase, etc. There is accumulating evidence supporting the idea that dietary nutraceuticals derived from commonly used fruits, vegetables, and spices have the ability to produce cancer chemopreventive activity by inducing Nrf2-mediated detoxifying enzymes. In this review, we discuss the importance of these nutraceuticals in cancer chemoprevention and summarize the role of dietary terpenoids in this respect. This approach was taken to accumulate the mechanistic function of these terpenoids to develop a comprehensive understanding of their direct and indirect roles in modulating the Keap1-Nrf2-ARE signaling system. [ABSTRACT FROM AUTHOR]

Peripheral nerve injuries are among those complicated medical conditions, which are still waiting for their highly effective first‐line therapies. In this study, the role of Calotropis procera crude leaves was evaluated at different doses for their effectiveness in improving functional recovery following sciatic nerve injury‐induced in the mouse model. Thirty‐two healthy albino mice were divided into four groups as Normal chow group (control, n = 8) and C. procera chow groups (50 mg/kg (n = 8), 100 mg/kg (n = 8) and 200 mg/kg (n = 8)). Behavioral analyses were performed to assess and compare improved functional recovery along with skeletal muscle mass measurement in all groups. Serum samples were analyzed for oxidative stress markers. Results showed that C. procera leaves at dose‐dependent manner showed statistically prominent (p <.05) increase in sensorimotor functions reclamation as confirmed by behavioral analyses along with muscle mass restoration and prominent decline in TOS and momentous increase in TAC along with the augmented activity of antioxidative enzymes. [ABSTRACT FROM AUTHOR]

Summary: Fragile X-related protein-1 (FXR1) gene is highly amplified in patients with ovarian cancer, and this amplification is associated with increased expression of both FXR1 mRNA and protein. FXR1 expression directly associates with the survival and proliferation of cancer cells. Surface sensing of translation (SUnSET) assay demonstrates that FXR1 enhances the overall translation in cancer cells. Reverse-phase protein array (RPPA) reveals that cMYC is the key target of FXR1. Mechanistically, FXR1 binds to the AU-rich elements (ARE) present within the 3′ untranslated region (3′UTR) of cMYC and stabilizes its expression. In addition, the RGG domain in FXR1 interacts with eIF4A1 and eIF4E proteins. These two interactions of FXR1 result in the circularization of cMYC mRNA and facilitate the recruitment of eukaryotic translation initiation factors to the translation start site. In brief, we uncover a mechanism by which FXR1 promotes cMYC levels in cancer cells.

Background: Severe sepsis, a major health problem worldwide, has become one of the leading causes of death in ICU patients. Further study on the pathogenesis and treatment of acute kidney injury (AKI) is of great significance to reduce high mortality rate of sepsis. In this study, the mechanism by which ubiquitin specific peptidase 10 (USP10) reduces sepsis-induced AKI was investigated. Ligation and perforation of cecum (CLP) was employed to establish C57BL/6 mouse models of sepsis. Hematoxylin-eosin (H&E) staining was performed to detect renal injury. The concentrations of serum creatinine (Cr), urea nitrogen (BUN) and cystatin C (Cys C) were determined using a QuantiChrom™ Urea Assay kit. RT-qPCR and western blot were conducted to assess the USP10 expression level. DHE staining was used to detect reactive oxygen species (ROS) levels. H2O2, MDA and SOD levels were assessed using corresponding colorimetric kits. Western blot was used to examine the expression levels of Bcl-2, Bax, cleaved caspase-3, Sirt6, Nrf2 and HO-1. MTT assay was used to determine cell viability, whereas TUNEL staining and flow cytometry were used to assess cell apoptosis. Results: In this study, we found that USP10 was decreased in CLP-induced mouse renal tissues. We identified that USP10 alleviated renal dysfunction induced by CLP. Moreover, USP10 was found to reduce oxidative stress, and abated LPS-induced renal tubular epithelial cell injury and apoptosis. Finally, we discovered that USP10 promoted activation of the NRF2/HO-1 pathway through SIRT6 and attenuated LPS-induced renal tubular epithelial cell injury. Conclusions: This study found that USP10 activates the NRF2/ARE signaling through SIRT6. USP10 alleviates sepsis-induced renal dysfunction and reduces renal tubular epithelial cell apoptosis and oxidative stress. [ABSTRACT FROM AUTHOR]

The integrated stress response (ISR) is a key cellular signaling pathway activated by environmental alterations that represses protein synthesis to restore homeostasis. To prevent sustained damage, the ISR is counteracted by the upregulation of growth arrest and DNA damage-inducible 34 (GADD34), a stress-induced regulatory subunit of protein phosphatase 1 that mediates translation reactivation and stress recovery. Here, we uncover a novel ISR regulatory mechanism that post-transcriptionally controls the stability of PPP1R15A mRNA encoding GADD34. We establish that the 3′ untranslated region of PPP1R15A mRNA contains an active AU-rich element (ARE) recognized by proteins of the ZFP36 family, promoting its rapid decay under normal conditions and stabilization for efficient expression of GADD34 in response to stress. We identify the tight temporal control of PPP1R15A mRNA turnover as a component of the transient ISR memory, which sets the threshold for cellular responsiveness and mediates adaptation to repeated stress conditions. [Display omitted] • Under normal conditions, TTP/Brf1 recognize PPP1R15A ARE promoting mRNA rapid decay • PPP1R15A mRNA turnover regulates the threshold at which the ISR is triggered • Cellular stress enhances PPP1R15A mRNA stability, favoring GADD34 translation • Decay of PPP1R15A mRNA enhances sensitivity to stress and antagonizes adaptation Magg et al. uncover a novel regulatory step of the integrated stress response at the level of PPP1R15A mRNA stability and establish AU-rich element-driven mRNA turnover as an important determinant controlling GADD34 expression, thereby shaping the dynamics of stress adaptation and sensitivity to repetitive stress exposure. [ABSTRACT FROM AUTHOR]

[Display omitted] • We identified novel Nrf2/ARE signaling pathway activators, natural (-)-galiellalactone 1 and its cyclohexene ring-truncated [3.3] bicyclic lactone variants 3 l and 3p. • (-)-Galiellalactone 1 and its variants 3l and 3p induced the ARE-mediated upregulation of downstream antioxidative and phase II detoxifying enzymes, HO-1 and NQO1. In particular, variant 3l remarkably increased the HO-1 expression. • (-)-Galiellalactone 1 and its variants 3l and 3p exhibited neuroprotective effects on the neuroblastoma SH-SY5Y cells against 6-OHDA-induced cytotoxic damages. Herein, (-)-galiellalactone 1 congeners responsible for the nuclear factor erythroid 2-related factor 2 (Nrf2)-activating neuroprotective effects were elucidated. Additionally, novel congener-based Nrf2 activators were identified using a drug repositioning strategy. (-)-Galiellalactone, which comprises a tricyclic lactone skeleton, significantly activates antioxidant response element (ARE)-mediated transcription in neuroblastoma SH-SY5Y cells. Interestingly, two cyclohexene-truncated [3.3] bicyclic lactone analogs, which possess an exocyclic α-methylene-γ-butyrolactone moiety, exhibited higher Nrf2/ARE transcriptional activities than the parent (-)-galiellalactone. We confirmed that the cyclohexene moiety embedding the [3.3] bicyclic lactone congener does not play the essential role of (-)-galiellalactone for Nrf2/ARE activation. Nrf2/ARE activation by novel analogs resulted in the upregulation of downstream antioxidative and phase II detoxifying enzymes, heme oxygenase-1, and NAD(P)H quinone oxidoreductase 1, which are closely related to the cytoprotective effects on neurodegenerative diseases. (-)-Galiellalactone and its [3.3] bicyclic variants 3l and 3p increased the expression of antioxidant genes and exhibited neuroprotective effects against 6-hydroxydopamine-mediated neurotoxicity in the neuroblastoma SH-SY5Y cell line. [ABSTRACT FROM AUTHOR]

We present arguments for an evolution in our understanding of how antioxidants in fruits and vegetables exert their health-protective effects. There is much epidemiological evidence for disease prevention by dietary antioxidants and chemical evidence that such compounds react in one-electron reactions with free radicals in vitro. Nonetheless, kinetic constraints indicate that in vivo scavenging of radicals is ineffective in antioxidant defense. Instead, enzymatic removal of nonradical electrophiles, such as hydroperoxides, in two-electron redox reactions is the major antioxidant mechanism. Furthermore, we propose that a major mechanism of action for nutritional antioxidants is the paradoxical oxidative activation of the Nrf2 (NF-E2-related factor 2) signaling pathway, which maintains protective oxidoreductases and their nucleophilic substrates. This maintenance of "nucleophilic tone," by a mechanism that can be called "para-hormesis," provides a means for regulating physiological nontoxic concentrations of the nonradical oxidant electrophiles that boost antioxidant enzymes, and damage removal and repair systems (for proteins, lipids, and DNA), at the optimal levels consistent with good health.

Background & aimsThe Keap1-Nrf2 signaling pathway regulates host cell defense responses against oxidative stress and maintains the cellular redox balance.MethodsWe investigated the function/molecular mechanisms by which Keap1-Nrf2 complex may influence liver ischemia/reperfusion injury (IRI) in a mouse model of hepatic cold storage (20h at 4°C) followed by orthotopic liver transplantation (OLT).ResultsThe Keap1 hepatocyte-specific knockout (HKO) in the donor liver ameliorated post-transplant IRI, evidenced by improved hepatocellular function and OLT outcomes (Keap1 HKO→Keap1 HKO; 100% survival), as compared with controls (WT→WT; 50% survival; p

The oxidative polymerizations of 2-methyl- m -phenylenediamine were performed to synthesize PMPDIA-A and PMPDIA-B polymers by H 2 O 2 (35% aqueous solution) oxidant in acid-catalyzed ethanol and basic aqueous medium. Furthermore, PMPDIA-E polymer of 2-methyl- m -phenylenediamine was enzymatically synthesized by HRP (horseradish peroxidase) enzyme and H 2 O 2 (35% aqueous solution) in dioxane/0.1 M phosphate tampon solution (pH = 7) mixture. The structures of 2-methyl- m -phenylenediamine and polymers were confirmed by FT-IR, NMR and UV–Vis spectrometer measurements. Molecular weight distributions, surface morphologies, thermal and fluorescence properties of polymers were determined from GPC, SEM, TG-DTA, DSC and fluorescence spectra analyses, respectively. CV and UV–Vis analyses were performed to determine the HOMO–LUMO energy levels, electrochemical (E g ′) and optical (E g) band gaps values of MPDIA, PMPDIA-E, PMPDIA-A and PMPDIA-B. The electrochemical and optical band gaps values of polymers were lower than MPDIA, because of their polyconjugated structures. According to GPC measurements of polymers, the weight average molecular weights were between 5400 and 10400 Da. The fluorescence quantum yield of PMPDIA-E was calculated to be 9.15% in DMSO solution. The adsorption of methylene blue (MB) from aqueous solution on PMPDIA-B was studied. Adsorption isotherms and equilibrium adsorption capacities were determined by the fitting of the experimental data to two well-known isotherm models: Langmuir and Freundlich. [ABSTRACT FROM AUTHOR]

Few studies have investigated the association between essential trace elements and oxidative stress in environmental media and populations with endemic arsenism caused by coal burning. Element contents and oxidative stress indicators were measured. Moreover, the expression of genes related to the nuclear factor E2-related factor 2 (Nrf2)/Kelch-like ECH-associated protein 1 (Keap1)-antioxidant response element (ARE) signaling pathway and Nrf2-ARE binding ability is detected. The results show that the contents of arsenic, copper, iron, and chromium were increased in environmental media from the arsenism area compared with the control area; however, the selenium content decreased. The arsenic, iron, chromium, and copper contents and the copper/zinc ratio were also increased in the arsenic-exposed population; however, the selenium content decreased. The results also show that the concentrations of arsenic, iron, and chromium and the copper/zinc ratio increased gradually with the severity of arsenism. However, selenium concentrations decreased gradually with the severity of arsenism. The contents of malondialdehyde, 8-hydroxyldeoxyguanosine, and protein carbonyl in plasma increased, while the levels of sulfhydryl, thioredoxin reductase (TrxR), glutathione peroxidase (Gpx), and superoxide dismutase 1 (SOD1) decreased. The mRNA expression of Keap1 and TrxR1 decreased in the blood, while the mRNA expression of Nrf2, GPx1, and SOD1 increased. Moreover, the Nrf2 protein content and Nrf2-ARE binding ability increased, and the Keap1 protein content decreased. In conclusion, our data suggest that the increased arsenic content in environmental media and populations was accompanied by abnormal levels of essential trace elements. Insufficient selenium intake, copper, and chromium overload and a high copper/zinc ratio might be some of the causes of arsenism, which might be related to the Nrf2/Keap1-ARE signaling pathway. [ABSTRACT FROM AUTHOR]

Brazilian red propolis reportedly has reactive oxygen species (ROS) scavenging effects in vitro, but the cellular mechanisms remain unclear. In the present study, the effects of an ethanol extract of Brazilian red propolis (EERP) on the Nrf2-ARE intracellular antioxidant pathway were examined in vitro and in vivo. EERP and its constituents transactivated the reporter gene through the ARE sequence and enhanced the expression of Nrf2-regulated genes in HEK293 cells. It also increased Nrf2 protein in the nucleus, which was partially inhibited by kinase inhibitors. Furthermore, EERP suppressed ROS generation and cytotoxicity induced by tert-butyl hydroperoxide. In vivo, orally administered EERP increased the expression of Nrf2-regulated genes in mice liver. These results suggest that EERP is a potential resource for preventing oxidative stress-related diseases as an Nrf2 inducer. "EERP activated the genes of Nrf2 and antioxidant enzymes in both HEK293 cells and the liver of mice administered EERP." [ABSTRACT FROM AUTHOR]

Vertebrate segmentation is regulated by the segmentation clock, a biological oscillator that controls periodic formation of somites, or embryonic segments, which give rise to many mesodermal tissue types. This molecular oscillator generates cyclic gene expression with the same periodicity as somite formation in the presomitic mesoderm (PSM), an area of mesenchymal cells that give rise to mature somites. Molecular components of the clock include the Hes/her family of genes that encode transcriptional repressors, but additional genes cycle. Cyclic gene transcripts are cleared rapidly, and clearance depends upon the pnrc2 (proline-rich nuclear receptor co-activator 2) gene that encodes an mRNA decay adaptor. Previously, we showed that the her1 3′UTR confers instability to otherwise stable transcripts in a Pnrc2-dependent manner, however, the molecular mechanism(s) by which cyclic gene transcripts are cleared remained largely unknown. To identify features of the her1 3′UTR that are critical for Pnrc2-mediated decay, we developed an array of transgenic inducible reporter lines carrying different regions of the 3′UTR. We find that the terminal 179 nucleotides (nts) of the her1 3′UTR are necessary and sufficient to confer rapid instability. Additionally, we show that the 3′UTR of another cyclic gene, deltaC (dlc) , also confers Pnrc2-dependent instability. Motif analysis reveals that both her1 and dlc 3′UTRs contain terminally-located Pumilio response elements (PREs) and AU-rich elements (AREs), and we show that the PRE and ARE in the last 179 ​nts of the her1 3′UTR drive rapid turnover of reporter mRNA. Finally, we show that mutation of Pnrc2 residues and domains that are known to facilitate interaction of human PNRC2 with decay factors DCP1A and UPF1 reduce the ability of Pnrc2 to restore normal cyclic gene expression in pnrc2 mutant embryos. Our findings suggest that Pnrc2 interacts with decay machinery components and cooperates with Pumilio (Pum) proteins and ARE-binding proteins to promote rapid turnover of cyclic gene transcripts during somitogenesis. [ABSTRACT FROM AUTHOR]

• iRIP-seq captures the RNA targets directly and indirectly bound by TTP. • Over-representation of AUUUA motif occurs not only in the 3'UTR but also in the intronic TTP binding peaks. • High frequencies of hnRNPA2/B1 and CELF1-binding motifs are presented in the 3'UTR and intronic TTP peaks. • The UGGAC motif overlapping with the m6A motif GGACU is over-represented in the CDS TTP peaks. • TTP upregulates RelB expression correlating with its multiple intronic and exonic sites in RelB. TTP up-regulates the expression of innate immunity genes in a RelB-dependent manner. Tristetraprolin (TTP) regulates inflammatory and immune responses by destabilizing target mRNAs via binding to their 3′-UTR AREs. We have recently reported that TTP preferentially up-regulates the expression level of innate immunity genes involved in the type I interferon-mediated signaling pathway and viral response in cancer cells. To elucidate the role of TTP-RNA interaction in TTP-mediated upregulation of gene expression, we performed iRIP-seq experiments to obtain the RNA interaction map consisting of direct and indirect binding sites of TTP in HeLa cells. We found substantial TTP binding signals in mRNA regions and the introns. ARE-motif AUUUA is over-represented in TTP binding peaks. Strikingly, AUUUA frequency is high both in 3′UTR and intronic regions, and the intronic peaks were more associated with TTP-regulated genes. Analysis of the over-represented motifs in TTP peaks revealed the high frequencies of UAGG and GUGUG motifs reported for hnRNPA2/B1 and CELF1 respectively in the 3′UTR and introns, and also the UGGAC motif overlapping with the m6A motif GGACU in the CDS regions. We further demonstrated that TTP binds to multiple intronic and exonic sites in the pre-mRNA/mRNA of the transcription factor RelB, correlating with the TTP-upregulated expression of RelB. TTP-up-regulated genes without a TTP binding site, but not those with, are highly enriched in innate immunity pathways and show higher tendency of harboring RelB binding sites in their promoter regions. These findings support a model in which TTP binding of RelB pre-mRNA/mRNA coordinates the RelB upregulation and activation of the innate immunity for antiviral response. [ABSTRACT FROM AUTHOR]

The peptide EDYGA was docked to the active pocket of Keap1 Kelch domain, three hydrogen bonds were found, the pocket of Keap1 Kelch domain are H bond donor, and EDYGA was hydrogen bond acceptor. • Peptide EDYGA was the most potent ARE-luciferase inducer. • EDYGA enhanced Nrf2 level by stabilizing Nrf2 by down regulating Keap1. • EDYGA was docked to the active pocket with three hydrogen bonds. The antioxidant response element (ARE) is a cis-acting enhancer sequence located in the region containing genes related to antioxidant and detoxification. Under oxidative stress, the induction of nuclear factor-E2-related factor 2 (Nrf2)/ARE is considered as a fundamental process involved in defending reactive oxygen species (ROS) and providing protection against toxic xenobiotics. In this study, we obtained seven antioxidant peptides from soft-shelled turtle and concluded that Glu-Asp-Tyr-Gly-Ala (EDYGA) is the most potent ARE-luciferase inducer. To gain fundamental insights into the role of EDYGA in oxidative stress, we evaluated the effects of EDYGA on the Nrf2/Keap1 system in HepG 2 cells. The results revealed that EDYGA modulated the Nrf2/ARE pathway by enhancing Nrf2 level through the stabilization of Nrf2, which was accomplished by a decrease in the level of Keap1. These actions eventually led to an increase in nuclear Nrf2 accumulation and ARE-binding activity. Moreover, silencing Nrf2 markedly reduced ARE-driven activity induced by EDYGA. Docking results proved that glutamate residues of peptide EDYGA directly bind to Arg 415 of Kelch domain receptor pocke. The results were helpful in understanding the antioxidant activity of peptides from soft-shelled turtle which have potential to be used in foods and drugs as functional ingredients. [ABSTRACT FROM AUTHOR]

Aldo-keto reductase (AKR) superfamily is responsible for preventing mammalian cells from the toxic and carcinogenic effect of different genotoxic and non-genotoxic chemicals by reducing them, though the inducibility of these genes are different in different species. The aim of this paper is to compare the gene regulation mechanisms of AKR superfamily genes in different species and to identify the conserved areas, which are responsible for gene regulations in the presence of antioxidant, toxicants, and non-genotoxic carcinogens. At the beginning of the analysis AKR genes found in different species were divided into two groups based on their amino acid sequence similarities. Comparison of AKR7A gene clusters between different species revealed that Human AKR7A2 has orthologues in mammalians like rat, mouse, pigs, and other primates. On the other hand, AKR7A3 has orthologues only in rat and AKR7L is present only in primates. All the genes of AKR superfamily have a trend to stay in clusters in mammalian chromosomes having repeated sequences in between them. Transcription start site analysis revealed that genes like human AKR7A2 and rat Akr7a4 do not have conventional promoter regions such as TATA box, CAAT box and have several GC-rich regions, whereas gene like Akr7a1 contains a TATA box 25 bp upstream of transcription start site instead of having CpG islands. Putative orthologous genes i.e., rat AKR7A4, human AKR7A2, and mouse AKR7A5 share more common features such as common transcription factor binding site for specificity protein 1 (SP1), GATA binding factor family, Selenocysteine tRNA gene transcription activating factor (STAF) zinc finger protein, Krüppel-like C2H2 zinc finger (HICF) protein, negative glucocorticoid response element (NGRE) etc. Similarly, genes like rat AKR7A1, human AKR7A3, and human AKR7L share common sequence and transcription factor binding sites. Among those, Nuclear factor erythroid 2-related factor 2 (Nrf2) is thought to be responsible for the inducibility of these genes in the presence of antioxidants. Our analysis revealed that AKR7A gene family consists of genes having a large number of variations in them. Some of these, such as AKR7A2 are housekeeping genes, on the other hand, genes like AKR7A3 are highly inducible in the presence of antioxidants because of the presence of Nrf2 binding site in their promoter. AKR7A1 has a different promoter than others and function of AKR7L gene is still unknown. [ABSTRACT FROM AUTHOR]

Dipeptidyl peptidase‐4 inhibitor (DPP‐4 inhibitor) such as sitagliptin has been presented as antidiabetic drugs and has numerous restorative advantages over different diseases; however, its defensive role against aflatoxin b1 (AFB1) liver toxicity has not been previously examined. Wistar rats (65 weeks, male) were utilized in the investigation. Animals were divided into five different groups (n = 10): control; AFB1; AFB1 + Sita (50); AFB1 + Sita (100); and Sita (100). Sitagliptin significantly (*p ≤.05, **p ≤.01, and ***p ≤.001) altered the levels of various serum liver enzymes (lactate dehydrogenase, alkaline phosphate, aspartate aminotransferase, and alanine aminotransferase). It decreased the concentration of an oxidative stress marker, that is, malondialdehyde and increased the level of antioxidant enzymes such as reduced glutathione, catalase, superoxide dismutase, and glutathione peroxidase in AFB1‐administered rats. It also improved the Nrf2 expression and HO‐1 level in AFB1‐intoxicated rats. This investigation discusses innovative evidence on the protective role of sitagliptin against AFB1‐induced hepatotoxicity in rats. [ABSTRACT FROM AUTHOR]

Background/Aims: To explore the effect of sulforaphane (SFN) treatment in rats through the induction of Stress Urinary Incontinence (SUI) via the Nrf2-ARE pathway. Methods: A total of 18 female rats (Sprague-Dawley) were assigned to three groups: a control group, an SUI group, and an SUI+SFN group (six rats per group). Rats in the treatment groups were induced via postpartum vaginal balloon dilation and bilateral ovariectomy. Rats in the SUI+SFN group were treated via intraperitoneal injection once per day for a total of one month. Urethral sphincter muscle histological was observed by HE and Masson staining. Peak voiding pressure and interval of micturition were measured by cystometry. Oxidative stress markers and protein expression in the Nrf2-ARE pathway were examined by immunohistochemical staining and western blotting. Results: Prolonged micturition interval and higher peak voiding pressure were observed in the SUI+SFN group. Disturbance of muscle morphology was ameliorated, muscle content was elevated, and collagen content was restrained in response to SFN treatment. The SOD, GSH-Px, and CAT activities were elevated in the SUI+SFN group compared to those in the control group. The level of cell apoptosis was decreased in SUI rats after SFN treatment; however, apoptosis was mainly located in the urethral mucosa instead of the muscle layer. SFN reduced the Bax/Bcl-2 expression ratio. Nrf2 and Nrf2 target antioxidant proteins were elevated in the SFN group. Conclusions: SFN was effective for SUI treatment via decreasing oxidative stress and activating the Nrf2-ARE pathway.

Background: Age-related macular degeneration (AMD) is the leading cause of blindness in elderly population in the developed world. Dysfunction of retinal pigment epithelium (RPE) likely triggers early AMD stages. The effect of Quercetin on the early AMD in-vitro model remained unclear. Methods: The effect of Quercetin in the cell viability was detected with CCK8 methods in control, CSE treated, and CSE with Quercetin treatment group. The apoptotic status in each group was detected with tunnel assay. The oxidative and inflammation biomarkers were detected by ELISA. The expression levels of Keap1/Nrf2/ARE in RPE cells were measured by western blot after pretreatment of Quercetin followed by CSE treatment. Results: It was found that Quercetin could improve the cell viability and decrease cellular apoptotic rate in the CSE treated RPE group. The expressions of inflammatory and apoptotic biomarkers were significantly decreased in Quercetin treatment group. Furthermore, Quercetin exerts protective effects via activation Keap1/Nrf2/ARE pathway in CSE treated RPE cells. Conclusions: Quercetin demonstrated significant protective effects in an in-vitro model of early AMD and it might be a new therapeutic strategy for the management of early AMD. [ABSTRACT FROM AUTHOR]

ROS, RNS, and carcinogenic metabolites generate excessive oxidative stress, which changes the basal cellular status and leads to epigenetic modification, genomic instability, and initiation of cancer. Epigenetic modification may inhibit tumor-suppressor genes and activate oncogenes, enabling cells to have cancer promoting properties. The nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that in humans is encoded by the NFE2L2 gene, and is activated in response to cellular stress. It can regulate redox homoeostasis by expressing several cytoprotective enzymes, including NADPH quinine oxidoreductase, heme oxygenase-1, UDP-glucuronosyltransferase, glutathione peroxidase, glutathione-S-transferase, etc. There is accumulating evidence supporting the idea that dietary nutraceuticals derived from commonly used fruits, vegetables, and spices have the ability to produce cancer chemopreventive activity by inducing Nrf2-mediated detoxifying enzymes. In this review, we discuss the importance of these nutraceuticals in cancer chemoprevention and summarize the role of dietary terpenoids in this respect. This approach was taken to accumulate the mechanistic function of these terpenoids to develop a comprehensive understanding of their direct and indirect roles in modulating the Keap1-Nrf2-ARE signaling system.