Glucose-capped gold nanoparticles (Glu-GNPs) have been used to improve cellular targeting and radio-sensitization. In this study, we explored the mechanism of Glu-GNP enhanced radiation sensitivity in radiation-resistant human prostate cancer cells. Cell survival and proliferation were measured using MTT and clonogenic assay. Flow cytometry with staining by propidium iodide (PI) was performed to study the cell cycle changes induced by Glu-GNPs, and western blotting was used to determine the expression of p53 and cyclin proteins that correlated to cell cycle regulation. With 2 Gy of ortho-voltage irradiation, Glu-GNP showed a 1.5-2.0 fold enhancement in growth inhibition when compared to x-rays alone. Comparing the cell cycle change, Glu-GNPs induced acceleration in the G0/G1 phase and accumulation of cells in the G2/M phase at 29.8% versus 18.4% for controls at 24 h. G2/M arrest was accompanied by decreased expression of p53 and cyclin A, and increased expression of cyclin B1 and cyclin E. In conclusion, Glu-GNPs trigger activation of the CDK kinases leading to cell cycle acceleration in the G0/G1 phase and accumulation in the G2/M phase. This activation is accompanied by a striking sensitization to ionizing radiation, which may have clinical implications.

Hypoxanthine is a mutagenic purine base that most commonly arises from the oxidative deamination of adenine. Damaged bases such as hypoxanthine are associated with carcinogenesis and cell death. This inevitable damage is counteracted by glycosylase enzymes, which cleave damaged bases from DNA. Alkyladenine DNA glycosylase (AAG) is the enzyme responsible for excising hypoxanthine from DNA in humans. In an effort to understand the intrinsic properties of hypoxanthine, we examined the gas-phase acidity and proton affinity using quantum mechanical calculations and gas-phase mass spectrometric experimental methods. In this work, we establish that the most acidic site of hypoxanthine has a gas-phase acidity of 332 +/- 2 kcal mol-1, which is more acidic than hydrochloric acid. We also bracket a less acidic site of hypoxanthine at 368 +/- 3 kcal mol-1. We measure the proton affinity of the most basic site of hypoxanthine to be 222 +/- 3 kcal mol-1. DFT calculations of these values are consistent with the experimental data. We also use calculations to compare the acidic and basic properties of hypoxanthine with those of the normal bases adenine and guanine. We find that the N9-H of hypoxanthine is more acidic than that of adenine and guanine, pointing to a way that AAG could discriminate damaged bases from normal bases. We hypothesize that AAG may cleave certain damaged nucleobases as anions and that the active site may take advantage of a nonpolar environment to favor deprotonated hypoxanthine as a leaving group versus deprotonated adenine or guanine. We also show that an alternate mechanism involving preprotonation of hypoxanthine is energetically less attractive, because the proton affinity of hypoxanthine is less than that of adenine and guanine. Last, we compare the acidity in the gas phase versus that in solution and find that a nonpolar environment enhances the differences in acidity among hypoxanthine, adenine, and guanine.

The gas phase acidity (DeltaH(acid) and DeltaG(acid)) and proton affinity (PA, and gas phase basicity (GB)) of adenine, guanine, and O(6)-methylguanine (OMG) have been examined using both theoretical (B3LYP/6-31+G*) and experimental (bracketing, Cooks kinetic) methods. We previously measured the acidity of adenine using bracketing methods; herein we measure the acidity of adenine by the Cooks kinetic method (DeltaH(acid) = 335 +/- 3 kcal mol(-1); DeltaG(acid) = 329 +/- 3 kcal mol(-1)). We also measured the PA/GB of adenine using both bracketing and Cooks methods (PA = 224 and 225 kcal mol(-1); GB = 216 and 217 kcal mol(-1)). Guanine is calculated to have several stable tautomers in the gas phase, in contrast to in solution, where the canonical tautomer predominates. Experimental measurements of gas phase guanine properties are difficult due to its nonvolatility; using electrospray and the Cooks kinetic method, we are able to measure a DeltaH(acid) of 335 +/- 3 kcal mol(-1) (DeltaG(acid) = 328 +/- 3 kcal mol(-1)). The proton affinity is 227 +/- 3 kcal mol(-1) (GB = 219 +/- 3 kcal mol(-1)). Comparison of these values to calculations indicates that we may have a mixture of the keto and enol tautomers under our conditions in the gas phase, although it is also possible that we only have the canonical form since in the Cooks method, we form the proton-bound dimers via electrospray of an aqueous solution, which should favor guanine in the canonical form. We also examined O(6)-methylguanine (OMG), a highly mutagenic damaged base that arises from the alkylation of guanine. Our calculations indicate that OMG may exist as both the "N9" (canonical) and "N7" (proton on N7 rather than N9) tautomers in the gas phase, as both are calculated to be within 3 kcal mol(-1) in energy. We have bracketed the acidity and proton affinity of OMG, which were previously unknown. The more acidic site of OMG has a DeltaH(acid) value of 338 +/- 3 kcal mol(-1) (DeltaG(acid) = 331 +/- 3 kcal mol(-1)). We have also bracketed the less acidic site (DeltaH(acid) = 362 +/- 3 kcal mol(-1), DeltaG(acid) = 355 +/- 3 kcal mol(-1)) and the PA (229 +/- 4 kcal mol(-1) (GB = 222 +/- 4 kcal mol(-1))). We confirmed these results through Cooks kinetic method measurements as well. Our ultimate goal is to understand the intrinsic reactivity of nucleobases; gas phase acidic and basic properties are of interest for chemical reasons and also possibly for biological purposes, since biological media can be quite nonpolar. We find that OMG is considerably less acidic at N9 than adenine and guanine and less basic at O6 than guanine; the biological implications of these differences are discussed.

As a globally transported pollutant, mercury (Hg) released from human activity and methylmercury (MeHg) in the food web are global concerns due to their increasing presence in the environment. In this study, we found that Hg released from municipal sewage into the environment in China is a substantial anthropogenic source based on mass sampling throughout China. In total, 160 Mg (140-190 Mg, from the 20th percentile to the 80th percentile) of Hg (THg) and 280 kg (240-330 kg) of MeHg were released from municipal sewage in China in 2015. The quantities of released THg and MeHg were the most concentrated in the coastal regions, especially in the East, North and South China regions. However, the per capita release of THg and MeHg was the highest in the Tibetan region, which is recognized as the cleanest region in China. THg released into aquatic environments was mitigated from 2001 to 2015 in China, but the amounts released into other sinks increased. This study provides the first picture of the release of Hg from municipal sewage into various sinks in China, and policy makers should pay more attention to the diversity and complexity of the sources and transport of Hg, which can lead to Hg accumulation in the food web and can threaten human health.

Modeling of the x-ray spectra of the Galactic superluminal jet sources GRS 1915+105 and GRO J1655-40 reveal a three-layered atmospheric structure in the inner region of their accretion disks. Above the cold and optically thick disk of a temperature 0.2-0.5 keV, there is a warm layer with a temperature of 1.0-1.5 keV and an optical depth around 10. Sometimes there is also a much hotter, optically thin corona above the warm layer, with a temperature of 100 keV or higher and an optical depth around unity. The structural similarity between the accretion disks and the solar atmosphere suggest that similar physical processes may be operating in these different systems.

We report herein a systematic mass spectrometric study of a series of thirty-one non-selfcomplementary, matched, DNA duplexes ranging in size from 5-to 12-mers. The purpose of this work is threefold: (1) to establish the viability of using mass spectrometry as a tool for examining solution phase stabilities of DNA duplexes; (2) to systematically assess gas-phase stabilities of DNA duplexes; and (3) to compare gas and solution phase stabilities in an effort to understand how media affects DNA stability. These fundamental issues are of importance both on their own, and also for harnessing the potential of mass spectrometry for biological applications. We have found that ion abundances do not always track with solution phase stability; GC content must be taken into account. Two duplexes with the same T m yet with differing GC content can yield different ion abundances. That is, if two duplexes have the exact same melting temperature, yet one has a higher GC content, the duplex with the higher GC content yields a higher ion abundance. It thus appears that not only is a GC base pair stronger than an AT base pair, but the relative strengths of each differ in the gas phase versus in solution, such that the electrospray process can differentiate between them. We also characterize the gas-phase stabilities of the duplexes, using collision-induced dissociation (CID) as a method to assess stability. We focus on two aspects of this CID experiment. One, we examine what factors appear to control whether the duplexes dissociate into single strands or covalently fragment; we are able to utilize a charge state normalization we coin "charge level" to compare our results with others' and establish generalities regarding dissociation versus fragmentation patterns. Two, we examine those duplexes that primarily dissociate and use CID to assess the gas-phase stabilities. We find that correlation of gas-phase to solution-phase stabilities is more likely to occur when duplexes of varying GC content are examined. Duplexes with the same GC content tend to have stabilities that do not parallel those in solution. We discuss these results in light of the different roles that hydrogen bonding and base stacking play in solution versus the gas phase. Ultimately, we apply what we learn to lend insight into the biological problem of how the carcinogenic, damaged nucleobase O 6 -methylguanine causes mutations. (J Am Soc Mass Spectrom 2006, 17, 1383-1395

A method to predict radiation-induced pneumonitis (RP) using an artificial neural network (ANN) was investigated. A retrospective study was applied to the clinical data from 142 patients who have been treated with three-dimensional conformal radiotherapy for tumors in the thoracic region. These data were classified, based on their treatment outcome, into two patient clusters: with RP (Np=26) and without RP (Np= 116). An ANN was designed as a classifier. To perform the classification, a patient-treatment outcome with RP was assigned a value of 1, and a patient treatment outcome without RP was assigned a value of -1. The input of the ANN was limited to the patient lung dose-volume data only. A volume vector (VD) that describes patient lung subvolumes receiving more than a set of threshold doses was used as the network input variable. A zero value was used as the threshold to set the output value into -1 or 1. Three ANNs (ANN_1, ANN_2, and ANN_3), each with three layers, were trained to perform this classification function and to show the effect of training data on the ANN performance. Radial basis function was applied as the hidden layer neuron activation function and a sigmoid function was selected as the output layer neuron function. Backpropagation with a conjugate gradient algorithm was used to train the network. ANN_1 was trained and tested by using the leave-one-out method. ANN_2 was trained by randomly selecting 2/3 of the patient data, and tested by the remaining 1/3 of the data. ANN_3 was trained by a user selecting 2/3 of the patient data, and tested by the remaining 1/3 of the data. The predictive accuracy was verified as the area under a receiver operator characteristic (ROC) curve. The correct classification rates of 73% for RP cases, and 99% for non-RP cases were obtained from ANN_1. The corresponding correct classification rates of 44% for RP cases, and 89% for non-RP cases were obtained from ANN_2. From the ANN_3 test phase, the corresponding correct classification rates of 55% for RP cases, and 95% non-RP cases were achieved. The area under ROC curve was 0.85+/-0.05, 0.68+/-0.10, and 0.81+/-0.09 for ANN_1, ANN _2, and ANN_3, respectively, within its asymmetric 95% confidence interval. The sensitivity was 95%, 57%, and 71%, and the specificity was 94%, 88%, and 90% for ANN_1, ANN_2, and ANN_3, respectively. Preliminary results suggest that the ANN approach provides a useful tool for the prediction of radiation-induced lung pneumonitis, using the patient lung dose-volume information.

No abstract