Redoxcitrinin, a biogenetic precursor of citrinin from marine isolate of fungus Penicillium sp

A chemical analysis of the fermentation of the marine-derived fungus Penicillium sp. led to the isolation of a biogenetic precursor of citrinin, redoxcitrinin (1), together with polyketide mycotoxins, phenol A (2), citrinin H2 (3), 4-hydroxymellein (4), citrinin (5), and phenol A acid (6). The structures of compounds 1-6 were determined on the basis of physicochemical data analyses. Among them, compounds 1-3 exhibited a potent radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) with IC50 values of 27.7, 23.4, and 27.2 microM, respectively.     

Affinity purification and characterization of a G-protein coupled receptor, Saccharomyces cerevisiae Ste2p

We present an example of expression and purification of a biologically active G-protein coupled receptor (GPCR) from yeast. An expression vector was constructed to encode the Saccharomyces cerevisiae GPCR alpha-factor receptor (Ste2p, the STE2 gene product) containing a 9-amino acid sequence of rhodopsin that served as an epitope/affinity tag. In the construct, two glycosylation sites and two cysteine residues were removed to aid future structural and functional studies. The receptor was expressed in yeast cells and was detected as a single band in a western blot indicating the absence of glycosylation. Ligand binding and signaling assays of the epitope-tagged, mutated receptor showed it maintained the full wild-type biological activity. For extraction of Ste2p, yeast membranes were solubilized with 0.5% n-dodecyl maltoside (DM). Approximately 120 microg of purified alpha-factor receptor was obtained per liter of culture by single-step affinity chromatography using a monoclonal antibody to the rhodopsin epitope. The binding affinity (K(d)) of the purified alpha-factor receptor in DM micelles was 28 nM as compared to K(d)=12.7 nM for Ste2p in cell membranes, and approximately 40% of the purified receptor was correctly folded as judged by ligand saturation binding. About 50% of the receptor sequence was retrieved from MALDI-TOF and nanospray mass spectrometry after CNBr digestion of the purified receptor. The methods described will enable structural studies of the alpha-factor receptor and may provide an efficient technique to purify other GPCRs that have been functionally expressed in yeast.     

Inhibition of mitochondrial respiration as a source of adaphostin-induced reactive oxygen species and cytotoxicity

Adaphostin is a dihydroquinone derivative that is undergoing extensive preclinical testing as a potential anticancer drug. Previous studies have suggested that the generation of reactive oxygen species (ROS) plays a critical role in the cytotoxicity of this agent. In this study, we investigated the source of these ROS. Consistent with the known chemical properties of dihydroquinones, adaphostin simultaneously underwent oxidation to the corresponding quinone and generated ROS under aqueous conditions. Interestingly, however, this quinone was not detected in intact cells. Instead, high performance liquid chromatography demonstrated that adaphostin was concentrated by up to 300-fold in cells relative to the extracellular medium and that the highest concentration of adaphostin (3000-fold over extracellular concentrations) was detected in mitochondria. Consistent with a mitochondrial site for adaphostin action, adaphostin-induced ROS production was diminished by >75% in MOLT-4 rho(0) cells, which lack mitochondrial electron transport, relative to parental MOLT-4 cells. In addition, inhibition of oxygen consumption was observed when intact cells were treated with adaphostin. Loading of isolated mitochondria to equivalent adaphostin concentrations caused inhibition of uncoupled oxygen consumption in mitochondria incubated with the complex I substrates pyruvate and malate or the complex II substrate succinate. Further analysis demonstrated that adaphostin had no effect on pyruvate or succinate dehydrogenase activity. Instead, adaphostin inhibited reduced decylubiquinone-induced cytochrome c reduction, identifying complex III as the site of inhibition by this agent. Moreover, adaphostin enhanced the production of ROS by succinate-charged mitochondria. Collectively, these observations demonstrate that mitochondrial respiration rather than direct redox cycling of the hydroquinone moiety is a source of adaphostin-induced ROS and identify complex III as a potential target for antineoplastic agents.     

Dosimetric and Monte Carlo verification of jaws-only IMRT plans calculated by the Collapsed Cone Convolution algorithm for head and neck cancers

AimThe aim of this study is to verify the Prowess Panther jaws-only intensity modulated radiation therapy (JO-IMRT) treatment planning (TP) by comparing the TP dose distributions for head-and-neck (H&N) cancer with the ones simulated by Monte Carlo (MC).BackgroundTo date, dose distributions planned using JO-IMRT for H&N patients were found superior to the corresponding three-dimensional conformal radiotherapy (3D-CRT) plans. Dosimetry of the JO-IMRT plans were also experimentally verified using an ionization chamber, MapCHECK 2, and Octavius 4D and good agreements were shown.Materials and methodsDose distributions of 15 JO-IMRT plans of nasopharyngeal patients were recalculated using the EGSnrc Monte Carlo code. The clinical photon beams were simulated using the BEAMnrc. The absorbed dose to patients treated by fixed-field IMRT was computed using the DOSXYZnrc. The simulated dose distributions were then compared with the ones calculated by the Collapsed Cone Convolution (CCC) algorithm on the TPS, using the relative dose error comparison and the gamma index using global methods implemented in PTW-VeriSoft with 3%/3 mm, 2%/2 mm, 1%/1 mm criteria.ResultsThere is a good agreement between the MC and TPS dose. The average gamma passing rates were 93.3 ± 3.1%, 92.8 ± 3.2%, 92.4 ± 3.4% based on the 3%/3 mm, 2%/2 mm, 1%/1 mm criteria, respectively.ConclusionsAccording to the results, it is concluded that the CCC algorithm was adequate for most of the IMRT H&N cases where the target was not immediately adjacent to the critical structures.     

Leucine Signals to mTORC1 via Its Metabolite Acetyl-Coenzyme A

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Tunable Hybrid Gap Surface Plasmon Polariton Waveguides with Ultralow Loss Deep-Subwavelength Propagation

Exploring hybrid gap surface plasmon polariton waveguides (HGSPPWs) is an important milestone in developing the next-generation, nanoscale integrated photonic circuit technology. To advance their potential applications, HGSPPWs are required to have tunable capability, highly reliable, simple fabrication process, and feasible integration. In this paper, we propose two tunable HGSPPWs fulfilling the requirements. The proposed HGSPPWs consist of a metallic wedge laterally coupled with a dielectric waveguide. The modal characteristics of HGSPPWs are investigated at the optical telecommunication wavelength, which shows the modal characteristics could be effectively controlled by tuning the key geometry parameters and structure of HGSPPWs. The propagation length could achieve the centimeter scale while maintaining the propagation mode size at the deep-subwavelength scale (~ λ²/10⁵). The studies on fabrication tolerance and waveguide crosstalk show their robust property for practical implementations. The effective tunable mechanism is also proposed and studied, which shows remarkable feasibility to realize multifunctional plasmon-based photonic components. Compared with the conventional HGSPPWs, the proposed HGSPPWs exhibit superior features in ultralow loss deep-subwavelength light guiding, are highly reliable, and are easy to integrate.

     

Quantification of Pseudocapacitive Contribution in Nanocage‐Shaped Silicon–Carbon Composite Anode

Pseudocapacitive materials have been highlighted as promising electrode materials to overcome slow diffusion‐limited redox mechanism in active materials, which impedes fast charging/discharging in energy storage devices. However, previously reported pseudocapacitive properties have been rarely used in lithium‐ion batteries (LIBs) and evaluation methods have been limited to those focused on thin‐film‐type electrodes. Hence, a nanocage‐shaped silicon–carbon composite anode is proposed with excellent pseudocapacitive qualities for LIB applications. This composite anode exhibits a superior rate capability compared to other Si‐based anodes, including commercial silicon nanoparticles, because of the higher pseudocapacitive contribution coming from ultrathin Si layer. Furthermore, unprecedent 3D pore design in cage shape, which prevents the particle scale expansion even after full lithiation demonstrates the high cycling stability. This concept can potentially be used to realize high‐power and high‐energy LIB anode materials.