No abstract

REV1 is a eukaryotic member of the Y-family of DNA polymerases involved in translesion DNA synthesis and genome mutagenesis. Recently, REV1 is also found to function in homologous recombination. However, it remains unclear how REV1 is recruited to the sites where homologous recombination is processed. Here, we report that loss of mammalian REV1 results in a specific defect in replication-associated gene conversion. We found that REV1 is targeted to laser-induced DNA damage stripes in a manner dependent on its ubiquitin-binding motifs, on RAD18, and on monoubiquitinated FANCD2 (FANCD2-mUb) that associates with REV1. Expression of a FANCD2-Ub chimeric protein in RAD18-depleted cells enhances REV1 assembly at laser-damaged sites, suggesting that FANCD2-mUb functions downstream of RAD18 to recruit REV1 to DNA breaks. Consistent with this suggestion we found that REV1 and FANCD2 are epistatic with respect to sensitivity to the double-strand break-inducer camptothecin. REV1 enrichment at DNA damage stripes also partially depends on BRCA1 and BRCA2, components of the FANCD2/BRCA supercomplex. Intriguingly, analogous to FANCD2-mUb and BRCA1/BRCA2, REV1 plays an unexpected role in protecting nascent replication tracts from degradation by stabilizing RAD51 filaments. Collectively these data suggest that REV1 plays multiple roles at stalled replication forks in response to replication stress.

Sacred lotus (Nelumbo nucifera) has long been used as a food source and ingredient for traditional herbal remedies. Plant parts contain neuroprotective agents that interact with specific targets to inhibit Alzheimer’s disease (AD). Organic solvents including methanol, ethyl acetate, hexane, and n-butanol, are widely employed for extraction of sacred lotus but impact food safety. Seed embryo, flower stalk, stamen, old leaf, petal, and leaf stalk of sacred lotus were extracted using hot water (aqueous extraction). The extractions were analyzed for their bioactive constituents, antioxidant and anti-AD properties as key enzyme inhibitory activities toward acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and β-secretase 1 (BACE-1). Results showed that the sacred lotus stamen exhibited significant amounts of phenolics, including phenolic acids and flavonoids, that contributed to high antioxidant activity via both single electron transfer (SET) and hydrogen atom transfer (HAT) mechanisms, with anti-AChE, anti-BChE, and anti-BACE-1 activities. To enhance utilization of other sacred lotus parts, a combination of stamen, old leaf and petal as the three sacred lotus plant components with the highest phenolic contents, antioxidant activities, and enzyme inhibitory properties was analyzed. Antagonist interaction was observed, possibly from flavonoids–flavonoids interaction. Further in-depth elucidation of this issue is required. Findings demonstrated that an aqueous extract of the stamen has potential for application as a functional food to mitigate the onset of Alzheimer’s disease.

Neuronal insulin resistance is a significant feature of Alzheimer’s disease (AD). Accumulated evidence has revealed the possible neuroprotective mechanisms of antidiabetic drugs in AD. Liraglutide, a glucagon-like peptide-1 (GLP-1) analog and an antidiabetic agent, has a benefit in improving a peripheral insulin resistance. However, the neuronal effect of liraglutide on the model of neuronal insulin resistance with Alzheimer’s formation has not been thoroughly investigated. The present study discovered that liraglutide alleviated neuronal insulin resistance and reduced beta-amyloid formation and tau hyperphosphorylation in a human neuroblostoma cell line, SH-SY5Y. Liraglutide could effectively reverse deleterious effects of insulin overstimulation. In particular, the drug reversed the phosphorylation status of insulin receptors and its major downstream signaling molecules including insulin receptor substrate 1 (IRS-1), protein kinase B (AKT), and glycogen synthase kinase 3 beta (GSK-3β). Moreover, liraglutide reduced the activity of beta secretase 1 (BACE-1) enzyme, which then decreased the formation of beta-amyloid in insulin-resistant cells. This indicated that liraglutide can reverse the defect of phosphorylation status of insulin signal transduction but also inhibit the formation of pathogenic Alzheimer’s proteins like Aβ in neuronal cells. We herein provided the possibility that the liraglutide-based therapy may be able to reduce such deleterious effects caused by insulin resistance. In view of the beneficial effects of liraglutide administration, these findings suggest that the use of liraglutide may be a promising therapy for AD with insulin-resistant condition.

Plukenetia volubilis or Inca peanut is a promising plant with high economic value. Its seeds can be pressed for oil production or roasted and served as a snack, while the dried leaves can be used to make a kind of tea. Although the oil from the cold‐pressed seeds has been proven to be safe for human consumption, little information is known about the other parts of the plant regarding safety. Thus, the aim of this study was to investigate the naturally occurring phytotoxins, including saponins, total alkaloids, and lectins in fresh and roasted Inca peanut seeds and leaves. In addition, cytotoxicity on several normal cell types including human peripheral blood mononuclear cells, human embryonic kidney cells, human hepatic stellate cells, and mouse fibroblasts as well as in vivo mutagenic properties was studied. This study showed that fresh Inca peanut seeds and leaves contain saponins, alkaloids, and lectins. However, roasting enables the reduction in alkaloids, saponins, and possibly lectins, suggesting that these phytotoxins become unstable under heat. Furthermore, Inca peanut seeds and leaves, especially after roasting, are safe to a variety of normal cell lines and do not induce DNA mutations in Drosophila expressing high biotransformation system. In conclusion, the data in this study indicated that high and chronic consumption of fresh seeds and leaves should be avoided. Heat processing should be applied before the consumption of Inca peanut seeds and leaves in order to reduce phytotoxins and potential health risks.

Short-wave ultraviolet light induces both mildly helix-distorting cyclobutane pyrimidine dimers (CPDs) and severely distorting (6–4) pyrimidine pyrimidone photoproducts ((6–4)PPs). The only DNA polymerase (Pol) that is known to replicate efficiently across CPDs is Polη, a member of the Y family of translesion synthesis (TLS) DNA polymerases. Phenotypes of Polη deficiency are transient, suggesting redundancy with other DNA damage tolerance pathways. Here we performed a comprehensive analysis of the temporal requirements of Y-family Pols ι and κ as backups for Polη in (i) bypassing genomic CPD and (6–4)PP lesions in vivo, (ii) suppressing DNA damage signaling, (iii) maintaining cell cycle progression and (iv) promoting cell survival, by using mouse embryonic fibroblast lines with single and combined disruptions in these Pols. The contribution of Polι is restricted to TLS at a subset of the photolesions. Polκ plays a dominant role in rescuing stalled replication forks in Polη-deficient mouse embryonic fibroblasts, both at CPDs and (6–4)PPs. This dampens DNA damage signaling and cell cycle arrest, and results in increased survival. The role of relatively error-prone Pols ι and κ as backups for Polη contributes to the understanding of the mutator phenotype of xeroderma pigmentosum variant, a syndrome caused by Polη defects.

This study investigated biological activities including antioxidative stress, anti‐inflammation, and anti‐insulin resistance of Anoectochilus burmannicus aqueous extract (ABE). The results showed abilities of ABE to scavenging DPPH and ABTS free radicals in a dose‐dependent manner. Besides, ABE significantly reduced nitric oxide (NO) production in the lipopolysaccharide (LPS)‐treated RAW 264.7 via inhibition of mRNA and protein expressions of nitric oxide synthase (iNOS). The LPS‐induced mRNA expressions of cyclooxygenase‐2 (COX‐2) and interleukin 1β (IL‐1β) were suppressed by ABE. Moreover, ABE exerted anti‐insulin resistance activity as it significantly improved the glucose uptake in tumor necrosis factor (TNF)‐α treated 3T3‐L1 adipocytes. In addition, ABE at the concentration of up to 200 μg/mL was not toxic to human peripheral blood mononuclear cells (PBMCs) and did not induce mutations. Finally, the results of our study suggest the potential use of A. burmannicus as anti‐inflammatory, anti‐insulin resistance agents, or food supplement for prevention of chronic diseases.

Anthocyanidins are bioactive compounds found mostly in colored plants and fruits. Consumption of anthocyanidin-rich foods has been shown to reduce the risk of diabetes. However, limited information is available regarding the inhibitory effect and interactions of anthocyanidins on α-glucosidase, the key enzyme that controls diabetes through degrading carbohydrate. Therefore, we used computational docking analysis to investigate the degree and type of inhibition by α-glucosidase, and the structural interactions of enzyme-selected anthocyanidins. The results suggested that anthocyanidins exhibit half maximal inhibitory concentration of 4∼55 μM; the strongest and weakest α-glucosidase inhibitors were delphinidin and malvidin, respectively. Indeed, delphinidin inhibits α-glucosidase in a mixed type, close to non-competitive manner with an inhibitory constant of 78 nM. Addition of a glycoside (glucoside or galactoside) at C3 on the C ring of delphinidin significantly decreased inhibitory activity, and addition of glycosides at C3 on the C ring and C5 on the A ring of delphinidin prevented all inhibitory activity. Molecular docking and free binding energy accurately confirmed the mode of inhibition determined by enzyme kinetics. These data will inform the use of alternative sources of anthocyanidins in functional foods and dietary supplements for prevention of diabetes. The results provide useful information for evaluating possible molecular models using anthocyanins/anthocyanidins as templates and α-glucosidase as the key enzyme in management of diabetes.