Monofunctional alkylating agents preferentially react at the N7 position of 2′-deoxyguanosine in duplex DNA. Methylated DNA, such as that produced by methyl methanesulfonate (MMS) and temozolomide, exists for days in organisms. The predominant consequence of N7-methyl-2′-deoxyguanosine (MdG) is widely believed to be abasic site (AP) formation via hydrolysis, a process that is slow in free DNA. Examination of MdG reactivity within nucleosome core particles (NCPs) provided two general observations. MdG depurination rate constants are reduced in NCPs compared with when the identical DNA sequence is free in solution. The magnitude of the decrease correlates with proximity to the positively charged histone tails, and experiments in NCPs containing histone variants reveal that positively charged amino acids are responsible for the decreased rate of abasic site formation from MdG. In addition, the lysine-rich histone tails form DNA-protein cross-links (DPCs) with MdG. Cross-link formation is reversible and is ascribed to nucleophilic attack at the C8 position of MdG. DPC and retarded abasic site formation are observed in NCPs randomly damaged by MMS, indicating that these are general processes. Histone-MdG cross-links were also detected by mass spectrometry in chromatin isolated from V79 Chinese hamster lung cells treated with MMS. The formation of DPCs following damage by a monofunctional alkylating agent has not been reported previously. These observations reveal the possibility that such DPCs may contribute to the cytotoxicity of monofunctional alkylating agents, such as MMS, N-methyl-N-nitrosourea, and temozolomide.

Abstract. Global warming or the increase of the surface and atmospheric temperatures of the Earth, is increasingly discernible in the polar, sub-polar and major land glacial areas. The Himalayan and Tibetan Plateau Glaciers, which are the largest glaciers outside of the Polar Regions, are showing a large-scale decrease of snow cover and an extensive glacial retreat. These glaciers such as Siachen and Gangotri are a major water resource for Asia as they feed major rivers such as the Indus, Ganga and Brahmaputra. Due to scarcity of ground measuring stations, the long-term observations of atmospheric temperatures acquired from the Microwave Sounding Unit (MSU) since 1979-2008 is highly useful. The lower and middle tropospheric temperature trend based on 30 years of MSU data shows warming of the Northern Hemisphere's midlatitude regions. The mean month-to-month warming (up to 0.048±0.026 • K/year or 1.44 • K over 30 years) of the mid troposphere (near surface over the high altitude Himalayas and Tibetan Plateau) is prominent and statistically significant at a 95% confidence interval. Though the mean annual warming trend over the Himalayas (0.016±0.005 • K/year), and Tibetan Plateau (0.008±0.006 • K/year) is positive, the month to month warming trend is higher (by 2-3 times, positive and significant) only over a period of six months (December to May). The factors responsible for the reversal of this trend from June to November are discussed here. The inequality in the magnitude of the warming trends of the troposphere between the western and easternCorrespondence to: A. K. Prasad (aprasad@chapman.edu) Himalayas and the IG (Indo-Gangetic) plains is attributed to the differences in increased aerosol loading (due to dust storms) over these regions. The monthly mean lowertropospheric MSU-derived temperature trend over the IG plains (dust sink region; up to 0.032±0.027 • K/year) and dust source regions (Sahara desert, Middle East, Arabian region, Afghanistan-Iran-Pakistan and Thar Desert regions; up to 0.068±0.033 • K/year) also shows a similar pattern of month-to-month oscillation and six months of enhanced and a statistically significant warming trend. The enhanced warming trend during the winter and pre-monsoon months (December-May) may accelerate glacial melt. The unequal distribution of the warming trend over the year is discussed in this study and is partially attributed to a number of controlling factors such as sunlight duration, CO 2 trends over the region (2003)(2004)(2005)(2006)(2007)(2008), water vapor and aerosol distribution.

Menthol is a common compound in pharmaceutical and commercial products and a popular additive to cigarettes. The molecular targets of menthol remain poorly defined. In this study we show an effect of menthol on the α7 subunit of the nicotinic acetylcholine (nACh) receptor function. Using a two-electrode voltage-clamp technique, menthol was found to reversibly inhibit α7-nACh receptors heterologously expressed in Xenopus oocytes. Inhibition by menthol was not dependent on the membrane potential and did not involve endogenous Ca2+-dependent Cl− channels, since menthol inhibition remained unchanged by intracellular injection of the Ca2+ chelator BAPTA and perfusion with Ca2+-free bathing solution containing Ba2+. Furthermore, increasing ACh concentrations did not reverse menthol inhibition and the specific binding of [125I] α-bungarotoxin was not attenuated by menthol. Studies of α7- nACh receptors endogenously expressed in neural cells demonstrate that menthol attenuates α7 mediated Ca2+ transients in the cell body and neurite. In conclusion, our results suggest that menthol inhibits α7-nACh receptors in a noncompetitive manner.

Menthol belongs to monoterpene class of a structurally diverse group of phytochemicals found in plant-derived essential oils. Menthol is widely used in pharmaceuticals, confectionary, oral hygiene products, pesticides, cosmetics, and as a flavoring agent. In addition, menthol is known to have antioxidant, anti-inflammatory, and analgesic effects. Recently, there has been renewed awareness in comprehending the biological and pharmacological effects of menthol. TRP channels have been demonstrated to mediate the cooling actions of menthol. There has been new evidence demonstrating that menthol can significantly influence the functional characteristics of a number of different kinds of ligand and voltage-gated ion channels, indicating that at least some of the biological and pharmacological effects of menthol can be mediated by alterations in cellular excitability. In this article, we examine the results of earlier studies on the actions of menthol with voltage and ligand-gated ion channels.

Advancement in our knowledge of deubiquitinases (DUBs) and their biological functions requires biochemical tools permitting interrogation of DUB activities under physiologically relevant conditions. Activity-based DUB probes (DUB ABPs) have been widely used in investigating the function and activity of DUBs. However, most ubiquitin (Ub)-based DUB ABPs are not cell-permeable, limiting their utility to purified proteins and cell lysates. Lysis of cells usually leads to dilution of the cytoplasm and disruption of the normal cellular organization, which may alter the activity of many DUBs and DUB complexes. Here, we report a new class of cell-permeable DUB ABPs that enable intracellular DUB profiling. We used a semisynthetic approach to generate modular ubiquitin-based DUB probes containing a reactive warhead for covalent trapping of DUBs with a catalytic cysteine. We employed cell-penetrating peptides (CPPs), particualrly cyclic polyarginine (cR10), to deliver the DUB ABPs into cells, as confirmed using live-cell fluorescence microscopy and DUB ABPs containing a fluorophore at the C-terminus of Ub. In comparison to TAT, enhanced intacellular delivery was observed through conjugation of a cyclic polyarginine (cR10) to the N-terminus of ubiquitin via a disulfide linkage. Using the new cell-permeable DUB ABPs, we carried out DUB profiling in intact HeLa cells, and identified active DUBs using immunocapture and label-free quantitative mass spectrometry. Additionally, we demonstrated that the cell-permeable DUB ABPs can be used in assessing the inhibition of DUBs by small-molecule inhibitors in intact cells. Our results indicate that cell-permeable DUB ABPs hold great promise in providing a better understanding of the cellular functions of DUBs and advancing drug discovery efforts targeting human DUBs.