A Comparison of Azacitidine and Decitabine Activities in Acute Myeloid Leukemia Cell Lines

Background

The cytidine nucleoside analogs azacitidine (AZA) and decitabine (DAC) are used for the treatment of patients with myelodysplastic syndromes and acute myeloid leukemia (AML). Few non-clinical studies have directly compared the mechanisms of action of these agents in a head-to-head fashion, and the agents are often viewed as mechanistically similar DNA hypomethylating agents. To better understand the similarities and differences in mechanisms of these drugs, we compared their in vitro effects on several end points in human AML cell lines.

Methodology/Principal Findings

Both drugs effected DNA methyltransferase 1 depletion, DNA hypomethylation, and DNA damage induction, with DAC showing equivalent activity at concentrations 2- to 10-fold lower than AZA. At concentrations above 1 µM, AZA had a greater effect than DAC on reducing cell viability. Both drugs increased the sub-G1 fraction and apoptosis markers, with AZA decreasing all cell cycle phases and DAC causing an increase in G2-M. Total protein synthesis was reduced only by AZA, and drug-modulated gene expression profiles were largely non-overlapping.

Conclusions/Significance

These data demonstrate shared mechanisms of action of AZA and DAC on DNA-mediated markers of activity, but distinctly different effects in their actions on cell viability, protein synthesis, cell cycle, and gene expression. The differential effects of AZA may be mediated by RNA incorporation, as the distribution of AZA in nucleic acid of KG-1a cells was 65∶35, RNA∶DNA.     

Protective Effect of Human Amniotic Fluid Stem Cells in an Immunodeficient Mouse Model of Acute Tubular Necrosis

Acute Tubular Necrosis (ATN) causes severe damage to the kidney epithelial tubular cells and is often associated with severe renal dysfunction. Stem-cell based therapies may provide alternative approaches to treating of ATN. We have previously shown that clonal c-kit^pos stem cells, derived from human amniotic fluid (hAFSC) can be induced to a renal fate in an ex-vivo system. Herein, we show for the first time the successful therapeutic application of hAFSC in a mouse model with glycerol-induced rhabdomyolysis and ATN. When injected into the damaged kidney, luciferase-labeled hAFSC can be tracked using bioluminescence. Moreover, we show that hAFSC provide a protective effect, ameliorating ATN in the acute injury phase as reflected by decreased creatinine and BUN blood levels and by a decrease in the number of damaged tubules and apoptosis therein, as well as by promoting proliferation of tubular epithelial cells. We show significant immunomodulatory effects of hAFSC, over the course of ATN. We therefore speculate that AFSC could represent a novel source of stem cells that may function to modulate the kidney immune milieu in renal failure caused by ATN.     

Vegetation classification of East China with multi-temporal NOAA-AVHRR data

East China lies in the subtropical monsoon climatic zone and is dominated by subtropical evergreen broad-leaved forests, a unique vegetation type mainly distributed in East Asia with the largest distribution in China. It is important to be able to monitor and estimate forest biomass and production, regional carbon storage, and global climate change impacts on these important vegetation types. In this paper, we used coarse resolution remote sensing data to identify the vegetation types in East China and developed a map of the spatial distribution of vegetation types in this region. Nineteen maximum normalized difference vegetation index (NDVI) composite images (acquisition time span of 7 months from February to August), which were derived from 10 days National Oceanographic and Atmospheric Administration (NOAA) Advanced Very High Resolution Radiometer (AVHRR) channel 1 and channel 2 observations, an unsupervised classification method, and the ISODATA algorithm were employed to identify the vegetation types. To reduce the dimensions of the dataset resulted in a total of 28 spectral clusters of land-cover of which two clusters were urban/bare soil and water, the images were processed using principal component analysis (PCA). The 26 remaining spectral clusters were merged into six vegetation types using the Chinese vegetation taxonomy system: evergreen broad-leaved forest, coniferous forest, bamboo forest, shrub-grass, aquatic vegetation, and agricultural vegetation. The spatial distribution and areal extent for the coniferous forests, shrub-grass, evergreen broad-leaved forests, and agricultural vegetation were calculated and compared with the Vegetation Atlas of China at a 1:1,000,000 scale. The spatial accuracy and the area accuracy for coniferous forests, shrub-grass, evergreen broad-leaved forests, and agricultural vegetation were 79.2%, 91.3%, 68.2% and 95.9% and 92.1%, 95.9%, 63.8% and 90.5%, respectively. The spatial accuracy and area accuracy of the bamboo forest were 28.7% and 96.5%, respectively; the spatial accuracy of aquatic vegetation was 69.6%, but there was a significant difference in its area accuracy because image acquisition did not cover the full year. Our study demonstrated the feasibility of using NOAA-AVHRR to identify the different vegetation types in the subtropical evergreen broad-leaved forest zone in East China. The spatial location of the six identified vegetation types agreed with the actual geographical distribution of the vegetation types in East China. __________ Translated from Acta Phytoecologica Sinica, 2005, 29(3): 436–443 [译自: 植物生态学报, 2005, 29(3): 436–443]     

Vegetation classification of East China with multi-temporal NOAA-AVHRR data

East China lies in the subtropical monsoon climatic zone and is dominated by subtropical evergreen broad-leaved forests,a unique vegetation type mainly distributed in East Asia with the largest distnbution in China.It is important to be able to monitor and estimate forest biomass and production,regional carbon storage,and global climate change impacts on these important vegetation types.In this paper,we used coarse resolution remote sensing data to identify the vegetation types in East China and developed a map of the spatial distribution of vegetation types in this region.Nineteen maximum normalized difference vegetation index(NDVI)composite images(acquisition time span of 7 months from February to August),which were derived from 10 days National Oceanographic and Atmospheric Administration(NOAA)Advanced Very High Resolution Radiometer(AVHRR)channel 1 and channel 2 observations,an unsupervised classification method,and the ISODATA algorithm were employed to identify the vegetation types.To reduce the dimensions of the dataset resulted in a total of 28 spectral clusters of land-cover of which two clusters were urban/bare soil and water,the images were processed using principal component analysis(PCA).The 26 remaining spectral clusters were merged into six vegetation types using the Chinese vegetation taxonomy system:evergreen broad-leaved forest,coniferous forest,bamboo forest,shrub-grass,aquatic vegetation,and agricultural vegetation.The spatial distribution and areal extent for the coniferous forests,shrub-grass,evergreen broad-leaved forests,and agricultural vegetation were calculated and comscale.The spatial accuracy and the area accuracy for coniferous forests,shrub-grass,evergreen broad-leaved forests,and agricultural vegetation were 79.2%,91.3%,68.2% and 95.9% and 92.1%,95.9%,63.8% and 90.5%,respectively.The spatial accuracy and area accuracy of the bamboo forest were 28.7% and 96.5%,respectively;the spatial accuracy of aquatic vegetation was 69.6%,but there was a significant difference in its area accuracy because image acquisition did not cover the full year.Our study demonstrated the fea sibility of using NOAA-AVHRR to identify the different vegetation types in the subtropical evergreen broad-leaved forest zone in East China.The spatial location of the six identified vegetation types agreed with the actual geo graphical distribution of the vegetation types in East China.     

An Eco-Friendly and Sustainable Process for Enzymatic Hydrolysis of Penicillin G in Cloud Point System

Enzymatic hydrolysis of penicillin G by immobilized penicillin acylase in a nonionic surfactant mediated cloud point system was presented. The effect of the operation parameters on equilibrium pH of this enzymatic hydrolysis process without pH control was examined. A relatively high equilibrium pH in cloud point system without pH control can be obtained. The feasibility of recycling utilization of the nonionic surfactant, a novel green solvent, was also investigated experimentally. Enzymatic hydrolysis of penicillin G in a discrete semi-batch mode, which simulates a semi-continuous process, envisages a completely eco-friendly, sustainable and efficient process for production of 6-aminopenicillanic acid.     

Colleters in Bathysa nicholsonii K. Schum. (Rubiaceae): ultrastructure, secretion protein composition, and antifungal activity

Colleters are secretory structures well distributed in many organs of Angiosperms. Ultrastructurally, the colleters secretory cell presents an enhanced endoplasmic reticulum, Golgi apparatus, and mitochondria. Secretion synthesis, transportation, and passage through outer cell wall is poorly characterized. This study characterized the anatomy and ultrastructure of BATHYSA NICHOLSONII (Rubiaceae) colleters and evaluated the presence of protein in the secretion and its antifungal property. Samples were collected and prepared according to usual techniques in light and electron microscopy, electrophoresis, and fungal growth inhibition assay. Colleters are of a standard type, cylindrical and elongated, formed by one secretory epidermal palisade layer, and a central axis formed by parenchymatic cells and a vascular trace. Epidermal cells have dense cytoplasm with abundant ribosome, a nucleus, enhanced endoplasmic reticulum and Golgi apparatus. The outer cell wall presented morphologically distinct layers. The presence of secretory cavities was noted in all outer cell wall extents. Secretion preparations analyzed by SDS-PAGE showed that B. NICHOLSONII secretion is a mixture of proteins with molecular masses covering a range of approximately 66 to 24 kDa. This preparation presented an inhibitory effect on the fungi spore growth.     

Arabidopsis PASTICCINO2 is an antiphosphatase involved in regulation of cyclin-dependent kinase A

PASTICCINO2 (PAS2), a member of the protein Tyr phosphatase-like family, is conserved among all eukaryotes and is characterized by a mutated catalytic site. The cellular functions of the Tyr phosphatase-like proteins are still unknown, even if they are essential in yeast and mammals. Here, we demonstrate that PAS2 interacts with a cyclin-dependent kinase (CDK) that is phosphorylated on Tyr and not with its unphosphorylated isoform. Phosphorylation of the conserved regulatory Tyr-15 is involved in the binding of CDK to PAS2. Loss of the PAS2 function dephosphorylated Arabidopsis thaliana CDKA;1 and upregulated its kinase activity. In accordance with its role as a negative regulator of the cell cycle, overexpression of PAS2 slowed down cell division in suspension cell cultures at the G2-to-M transition and early mitosis and inhibited Arabidopsis seedling growth. The latter was accompanied by altered leaf development and accelerated cotyledon senescence. PAS2 was localized in the cytoplasm of dividing cells but moved into the nucleus upon cell differentiation, suggesting that the balance between cell division and differentiation is regulated through the interaction between CDKA;1 and the antiphosphatase PAS2.     

Surface photochemistry: benzophenone as a probe for the study of silica and reversed-phase silica surfaces.

This work reports the use of benzophenone, a very well characterized probe, to study new hosts: two reversed-phase silicas. Laser-induced room temperature luminescence of argon purged solid powdered samples of benzophenone adsorbed onto the two different reversed-phase silicas, RP-18 and RP-8, revealed the existence of a low energy emission band in contrast with the benzophenone adsorbed on 60 A pore silica, where only triplet benzophenone emits. This low energy emission band was identified as the fluorescence of the ketyl radical of benzophenone, which is formed as the result of a hydrogen atom abstraction reaction of the triplet excited benzophenone from the alkyl groups of the surface of the reversed silicas. Such emission does not exist for benzophenone adsorbed onto 60 A pore silica. Room temperature phosphorescence was obtained in argon purged samples for all the surfaces under use. The decay times of the benzophenone emission vary greatly with the alkylation of the silica surface when compared with "normal" silica surface. A lifetime distributions analysis has shown that the shortest lifetimes for the benzophenone emission exist in the former case. Triplet-triplet absorption of benzophenone was detected in all cases and is the predominant absorption in the case of 60 A pore silica, while benzophenone ketyl radical formation occurs in the case of the reversed silicas. Diffuse reflectance laser flash photolysis and gas chromatography-mass spectrometry techniques provided complementary information, the former about transient species and the latter regarding the final products formed after laser irradiation, both at 266 nm or 355 nm. Product analysis and identification show that the degradation photoproducts are dependent on the excitation wavelength, the photochemistry being much more rich and complex in the 266 nm excitation case, where an alpha-cleavage reaction occurs. A detailed mechanistic analysis is proposed.     

Effects of replacing the unreliable cDNA microarray measurements on the disease classification based on gene expression profiles and functional modules

MOTIVATION: Microarrays datasets frequently contain a large number of missing values (MVs), which need to be estimated and replaced for subsequent data mining. The focus of the paper is to study the effects of different MV treatments for cDNA microarray data on disease classification analysis. RESULTS: By analyzing five datasets, we demonstrate that among three kinds of classifiers evaluated in this study, support vector machine (SVM) classifiers are robust to varied MV imputation methods [e.g. replacing MVs by zero, K nearest-neighbor (KNN) imputation algorithm, local least square imputation and Bayesian principal component analysis], while the classification and regression tree classifiers are sensitive in terms of classification accuracy. The KNNclassifiers built on differentially expressed genes (DEGs) are robust to the varied MV treatments, but the performances of the KNN classifiers based on all measured genes can be significantly deteriorated when imputing MVs for genes with larger missing rate (MR) (e.g. MR > 5%). Generally, while replacing MVs by zero performs relatively poor, the other imputation algorithms have little difference in affecting classification performances of the SVM or KNN classifiers. We further demonstrate the power and feasibility of our recently proposed functional expression profile (FEP) approach as means to handle microarray data with MVs. The FEPs, which are derived from the functional modules that are enriched with sets of DEGs and thus can be consistently identified under varied MV treatments, achieve precise disease classification with better biological interpretation. We conclude that the choice of MV treatments should be determined in context of the later approaches used for disease classification. The suggested exclusion criterion of ignoring the genes with larger MR (e.g. >5%), while justifiable for some classifiers such as KNN classifiers, might not be considered as a general rule for all classifiers.     

A new UV-visible confocal laser scanning microspectrofluorometer designed for spectral cellular imaging

With the aim to perform spectroscopic studies and spectral images inside living cells, a microspectrofluorometer has been designed for two-dimensional spectral imaging in the visible and in the near-UV region. The main advantage of the device relies on its ability to scan the laser beam along one direction of the sample. This scanning is optically coupled with one direction of a bidimensional detector, allowing an instantaneous recording of a one-dimensional spectral image. The overall scanning of the sample is achieved by means of submicrometric displacements of the stage in the perpendicular direction. The main characteristics and performances of the microspectrofluorometer in terms of sensitivity (detection of a few molecules), spatial resolution (0.5 x 0.5 x 1 microm), and spectral resolution (1 nm) are presented. Finally, applications of this new apparatus concerning in situ localization and spectral characterization of two dyes are shown with Drosophila salivary glands (ethidium bromide) and T47D tumor cells (Hoechst 33342).