DNA Rereplication Is Susceptible to Nucleotide-Level Mutagenesis.

The sources of genome instability, a hallmark of cancer, remain incompletely understood. One potential source is DNA rereplication, which arises when the mechanisms that prevent the reinitiation of replication origins within a single cell cycle are compromised. Using the budding yeast Saccharomyces cerevisiae, we previously showed that DNA rereplication is extremely potent at inducing gross chromosomal alterations and that this arises in part because of the susceptibility of rereplication forks to break. Here, we examine the ability of DNA rereplication to induce nucleotide-level mutations. During normal replication these mutations are restricted by three overlapping error-avoidance mechanisms: the nucleotide selectivity of replicative polymerases, their proofreading activity, and mismatch repair. Using lys2InsEA14 , a frameshift reporter that is poorly proofread, we show that rereplication induces up to a 30× higher rate of frameshift mutations and that this mutagenesis is due to passage of the rereplication fork, not secondary to rereplication fork breakage. Rereplication can also induce comparable rates of frameshift and base-substitution mutations in a more general mutagenesis reporter CAN1, when the proofreading activity of DNA polymerase ε is inactivated. Finally, we show that the rereplication-induced mutagenesis of both lys2InsEA14 and CAN1 disappears in the absence of mismatch repair. These results suggest that mismatch repair is attenuated during rereplication, although at most sequences DNA polymerase proofreading provides enough error correction to mitigate the mutagenic consequences. Thus, rereplication can facilitate nucleotide-level mutagenesis in addition to inducing gross chromosomal alterations, broadening its potential role in genome instability

The RootScope: A Simple High-Throughput Screening System For Quantitating Gene Expression Dynamics In Plant Roots

Background: High temperature stress responses are vital for plant survival. The mechanisms that plants use to sense high temperatures are only partially understood and involve multiple sensing and signaling pathways. Here we describe the development of the RootScope, an automated microscopy system for quantitating heat shock responses in plant roots.Results: The promoter of Hsp17.6 was used to build a Hsp17.6(p):GFP transcriptional reporter that is induced by heat shock in Arabidopsis. An automated fluorescence microscopy system which enables multiple roots to be imaged in rapid succession was used to quantitate Hsp17.6p: GFP response dynamics. Hsp17.6(p):GFP signal increased with temperature increases from 28 degrees C to 37 degrees C. At 40 degrees C the kinetics and localization of the response are markedly different from those at 37 degrees C. This suggests that different mechanisms mediate heat shock responses above and below 37 degrees C. Finally, we demonstrate that Hsp17.6(p):GFP expression exhibits wave like dynamics in growing roots.Conclusions: The RootScope system is a simple and powerful platform for investigating the heat shock response in plants

Composted tobacco waste increases the yield and organoleptic quality of leaf mustard

Tobacco (Nicotiana tabacum) waste is produced in many countries and is phytotoxic due to the alkaloid content; in Vietnam the waste is usually burned causing air pollution. We composted tobacco waste with chicken manure in different proportions—1 t of waste ± accelerant (C1 and C2); 0.8 t of waste + 0.2 t of chicken manure ± accelerant (C3 and C4); and 0.7 t of waste + 0.3 t of chicken manure ± accelerant (C5 and C6)—for 30 d in covered heaps. Three mixtures containing the accelerant (C2, C4, and C6) reached temperatures of ∼55 °C, that 5s, hot enough to suppress disease and weeds. Composting decreased the alkaloid content from ∼6,000 to ∼200 mg kg−1, and C4 with a C/N ratio of 19:1, was used in a field trial. The compost treatments (0, 10, 15, and 20 t ha−1) were combined fertilizer with phosphorus (40 kg ha−1), nitrogen (60 kg ha−1) and potassium (90 kg ha−1) for leaf mustard (Brassica integrifolia). The yield increased from ∼17 to ∼29 t ha−1 with the amount of compost applied, and the nitrate concentration decreased concomitantly from ∼67 to ∼42 mg NO3–N kg−1 fresh weight, presumably due to ongoing composting (nitrogen drawdown). Organoleptic evaluation showed a preference for the crops grown with the compost amendments. Whether remains to be seen whether one-off compost applications >20 t ha−1 and repeated, large applications provide additional, long-term production benefits, or if the benefits may be outweighed by the accumulation of persistent, phytotoxic alkaloids

Chemokine and inflammatory cytokine changes during chronic wound healing

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75761/1/j.1524-475X.1997.50405.x.pd

An experiment in Interactive Retrieval for the lifelog moment retrieval task at imageCLEFlifelog2020.

The development of technology has led to an increase in mobile devices’ use to keep track of individual daily activities, as known as Lifelogging. Lifelogging has raised many research challenges, one of which is how to retrieve a specific moment in response to a user’s information need. This paper presents an efficient interactive search engine for large multimodal lifelog data which is evaluated in the ImageCLEFlifelog2020 Lifelog Moment Retrieval task (LMRT). The system is the modified version of the Myscéal demonstrator used in the Lifelog Search Challenge 2020, with the addition of visual similarity and a new method of visualising results. In interactive experimentation, our system achieved an F1@ 10 score of 0.48 in the official submission but can be significantly improved by implementing a number of post-processing steps

Chronic inhibition of tumor cell-derived VEGF enhances the malignant phenotype of colorectal cancer cells

Abstract Background Vascular endothelial growth factor-a (VEGF)-targeted therapies have become an important treatment for a number of human malignancies. The VEGF inhibitors are actually effective in several types of cancers, however, the benefits are transiently, and the vast majority of patients who initially respond to the therapies will develop resistance. One of possible mechanisms for the acquired resistance may be the direct effect(s) of VEGF inhibitors on tumor cells expressing VEGF receptors (VEGFR). Thus, we investigated here the direct effect of chronic VEGF inhibition on phenotype changes in human colorectal cancer (CRC) cells. Methods To chronically inhibit cancer cell-derived VEGF, human CRC cell lines (HCT116 and RKO) were chronically exposed (2 months) to an anti-VEGF monoclonal antibody (mAb) or were disrupted the Vegf gene (VEGF-KO). Effects of VEGF family members were blocked by treatment with a VEGF receptor tyrosine kinase inhibitor (VEGFR-TKI). Hypoxia-induced apoptosis under VEGF inhibited conditions was measured by TUNEL assay. Spheroid formation ability was assessed using a 3-D spheroid cell culture system. Results Chronic inhibition of secreted/extracellular VEGF by an anti-VEGF mAb redundantly increased VEGF family member (PlGF, VEGFR1 and VEGFR2), induced a resistance to hypoxia-induced apoptosis, and increased spheroid formation ability. This apoptotic resistance was partially abrogated by a VEGFR-TKI, which blocked the compensate pathway consisted of VEGF family members, or by knockdown of Vegf mRNA, which inhibited intracellular function(s) of all Vegf gene products. Interestingly, chronic and complete depletion of all Vegf gene products by Vegf gene knockout further augmented these phenotypes in the compensate pathway-independent manner. These accelerated phenotypes were significantly suppressed by knockdown of hypoxia-inducible factor-1α that was up-regulated in the VEGF-KO cell lines. Conclusions Our findings suggest that chronic inhibition of tumor cell-derived VEGF accelerates tumor cell malignant phenotypes.http://deepblue.lib.umich.edu/bitstream/2027.42/112625/1/12885_2012_Article_3866.pd

Antibacterial hydrogel containing Piper betle L. extract for acne treatment, an ex vivo investigation

The current treatments of acne vulgaris and acne-like disorders such as gram-negative folliculitis possess lots of unwanted side effects. Thus, alternative approach of utilizing natural plant extracts, specifically Piper betle L., have gained much attention. To this end, this work developed, characterized, and ex vivo evaluated novel antibiotics hydrogels containing P. betle L. leaf extract for acne treatment. Firstly, the design of experiments (DoE) D-optimal method was successfully developed, optimized, and validated, to investigate the relationship between P. betle L. extraction conditions and the extract properties. Secondly, the best extract was encapsulated in the hydrogel formulations composed of carbopol 940, propylene glycol, and cocamidopropyl betaine. Finally, the hydrogel was ex vivo determined its antibacterial activity on bacteria isolated from 15 patient acne samples. The optimal extraction condition being an extraction solvent/plant weight ratio of 4.034:1, an extraction time of 2.147 h, and a water extract volume of 91.4 mL. This condition yielded an extract total phenolic content of 3.337±0.034 g GAE/g, and minimum inhibitory concentrations of 32 µg/mL and 128 µg/mL on Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922, respectively. The hydrogel possessed suitable properties for a topical medication, including a viscosity of 6800 cps, a pH of 7.0, and a good foaming ability, at both 10°C, 25°C, and 40°C. The hydrogel showed higher antibacterial activity than the positive controls in both gram-positive and gram-negative bacteria. Conclusively, the hydrogel could become a potential pharmaceutical product for acne treatment