Commenced in January 2007
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Edition: International
Paper Count: 22

Search results for: acetylation

22 Theoretical Study of Acetylation of P-Methylaniline Catalyzed by Cu²⁺ Ions

Authors: Silvana Caglieri

Abstract:

Theoretical study of acetylation of p-methylaniline catalyzed by Cu2+ ions from the analysis of intermediate of the reaction was carried out. The study of acetylation of amines is of great interest by the utility of its products of reaction and is one of the most frequently used transformations in organic synthesis as it provides an efficient and inexpensive means for protecting amino groups in a multistep synthetic process. Acetylation of amine is a nucleophilic substitution reaction. This reaction can be catalyzed by Lewis acid, metallic ion. In reaction mechanism, the metallic ion formed a complex with the oxygen of the acetic anhydride carbonyl, facilitating the polarization of the same and the successive addition of amine at the position to form a tetrahedral intermediate, determining step of the rate of the reaction. Experimental work agreed that this reaction takes place with the formation of a tetrahedral intermediate. In the present theoretical work were investigated the structure and energy of the tetrahedral intermediate of the reaction catalyzed by Cu2+ ions. Geometries of all species involved in the acetylation were made and identified. All of the geometry optimizations were performed by the method at the DFT/B3LYP level of theory and the method MP2. Were adopted the 6-31+G* basis sets. Energies were calculated using the Mechanics-UFF method. Following the same procedure it was identified the geometric parameters and energy of reaction intermediate. The calculations show 61.35 kcal/mol of energy for the tetrahedral intermediate and the energy of activation for the reaction was 15.55 kcal/mol.

Keywords: amides, amines, DFT, MP2

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21 Effects of Valproate on Vascular Endothelial Growth Factor in the Retina Associated with Choroidal Neovascularization

Authors: Zhang Zhenzhen

Abstract:

Valproate (VPA) is commonly used in the treatment of bipolar disorder and epilepsy. The mechanism is complicated, including its ability to inhibit histone deacetylases (HDACs). Here, we show that VPA attenuated VEGF gene expression and the morphological changes in choroidal neovascularization (CNV) induced by photocoagulation in retina. C57BL/6 mice were injected subcutaneously at 300mg/kg twice daily with VPA before insult. Vascular endothelial growth factor (VEGF)-A and VEGF-B were examined in the eyes of VPA-treated mice and in human retinal pigment epithelial cell lines (ARPE-19) exposed to VPA. In addition, CNV was induced by photocoagulation in mice injected with VPA, and the volume of CNV was compared by fluorescence-labeled choroidal flat mount. Morphological changes were analyzed on stained histological sections. Western blot analysis was used to determine protein levels of VEGF-A and VEGF-B, and acetylation of histone H3 in each group. VPA injected intraperitoneally attenuated the VEGF-A and VEGF-B expression in the retina, accompanied by the hyperacetylation of retina tissue, indicating that VPA acts directly on retina tissues through acetylation to reduce the expression of VEGF. VPA also attenuated the VEGF-A mRNA expression in the retinal pigment epithelium showed by immunohistochemistry. Moreover, the administration of VPA significantly attenuated photocoagulation-induced CNV in mice. These results demonstrate that VPA attenuated VEGF production in retina associated with choroidal neovascularization possibly via the HDAC inhibition.

Keywords: retina, acetylation, chorodial neovascularization, vascular endothelial growth factor

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20 Advances in Natural Fiber Surface Treatment Methodologies for Upgradation in Properties of Their Reinforced Composites

Authors: G. L. Devnani, Shishir Sinha

Abstract:

Natural fiber reinforced polymer composite is a very attractive area among the scientific community because of their low cost, eco-friendly and sustainable in nature. Among all advantages there are few issues which need to be addressed, those issues are the poor adhesion and compatibility between two opposite nature materials that is fiber and matrix and their relatively high water absorption. Therefore, natural fiber modifications are necessary to improve their adhesion with different matrices. Excellent properties could be achieved with the surface treatment of these natural fibers ultimately leads to property up-gradation of their reinforced composites with different polymer matrices. Lot of work is going on to improve the adhesion between reinforced fiber phase and polymer matrix phase to improve the properties of composites. Researchers have suggested various methods for natural fiber treatment like silane treatment, treatment with alkali, acetylation, acrylation, maleate coupling, etc. In this study a review is done on the different methods used for the surface treatment of natural fibers and what are the advance treatment methodologies for natural fiber surface treatment for property improvement of natural fiber reinforced polymer composites.

Keywords: composites, acetylation, natural fiber, surface treatment

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19 Posttranslational Modifications of Histone H3 in Tumor Tissue Isolated from Silver and Gold Nanoparticles Treated Mice

Authors: Lucyna Kapka-Skrzypczak, Barbara Sochanowicz, Magdalena Matysiak-Kucharek, Magdalena Czajka, Krzysztof Sawicki, Marcin Kruszewski

Abstract:

Due to the strong antimicrobial activity silver nanoparticles (AgNPs) are widely used in various medical and general applications, among others, in cosmetics, odour resistant textiles, etc. The aim of this study was to compare effect of AgNPs and gold NPs (AuNPs) on histones posttranslational modifications. Histone molecule posttranscriptional modifications are responsible for chromatin compaction and repackaging. In this study, BALB/c mice were inoculated with murine mammary carcinoma 4T1 cells and treated with AgNPs coated with citrate (AgNPs(cit) or PEG (AgNPs(PEG), or AuNPs. Thereafter the histone H3 acetylation on Lys9 and H3 methylation on Lys4, Lys9, Lys29 was investigated. All NPs tested decreased H3 methylation, while no effect was observed for H3 acetylation. Modification of histone H3 methylation dependent on type of NPs used its coating, site of methylation and treatment used. Conclusion, epigenetic effects of nanomaterials depend on nanomaterial composition, its coating, and way of application. This work was supported by National Science Centre grant No. 2014/15/B/NZ7/01036 (MK, LKS, MMK, MC, KS), statutory funding for INTC (BS).

Keywords: gold nanoparticles, histone, methylation, silver nanoparticles

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18 Aberrant Acetylation/Methylation of Homeobox (HOX) Family Genes in Cumulus Cells of Infertile Women with Polycystic Ovary Syndrome (PCOS)

Authors: P. Asiabi, M. Shahhoseini, R. Favaedi, F. Hassani, N. Nassiri, B. Movaghar, L. Karimian, P. Eftekhariyazdi

Abstract:

Introduction: Polycystic Ovary Syndrome is a common gynecologic disorder. Many factors including environment, metabolism, hormones and genetics are involved in etiopathogenesis of PCOS. Of genes that have altered expression in human reproductive system disorders are HOX family genes which act as transcription factors in regulation of cell proliferation, differentiation, adhesion and migration. Since recent evidences consider epigenetic factors as causative mechanisms of PCOS, evaluation of association between known epigenetic marks of acetylation/methylation of histone 3 (H3K9ac/me) with regulatory regions of these genes can represent better insight about PCOS. In the current study, cumulus cells (CCs) which have critical roles during folliculogenesis, oocyte maturation, ovulation and fertilization were aimed to monitor epigenetic alterations of HOX genes. Material and methods: CCs were collected from 20 PCOS patients and 20 fertile women (18-36 year) with male infertility problems referred to the Royan Institute to have ICSI under GnRH antagonist protocol. Informed consents were obtained from the participants. Thirty six hours after hCG injection, ovaries were punctured and cumulus oocyte complexes were dissected. Soluble chromatin were extracted from CCs and Chromatin Immune precipitation (ChIP) coupled with Real Time PCR was performed to quantify the epigenetic marks of histone H3K9 acetylation/methylation (H3K9ac/me) on regulatory regions of 15 members of HOX genes from A-D subfamily. Results: Obtained data showed significant increase of H3K9ac epigenetic mark on regulatory regions of HOXA1, HOXB2, HOXC4, HOXD1, HOXD3 and HOXD4 (P < 0.01) and HOXC5 (P < 0.05) and also significant decrease of H3K9ac into regulatory regions of HOXA2, HOXA4, HOXA5, HOXB1 and HOXB5 (P < 0.01) and HOXB3 (P<0.05) in PCOS patients vs. control group. On the other side, there was a significant decrease in incorporation of H3K9me level on regulatory region of HOXA2, HOXA3, HOXA4, HOXA5, HOXB3 and HOXC4 (P≤0.01) and HOXB5 (P < 0.05) in PCOS patients vs. control group. This epigenetic mark (H3K9me2) has significant increase on regulatory region of HOXB1, HOXB2, HOXC5, HOXD1, HOXD3 and HOXD4 (P ≤ 0.01) and HOXB4 (P < 0.05) in patients vs. control group. There were no significant changes in acetylation/methylation levels of H3K9 on regulatory regions of the other studied genes. Conclusion: Current study suggests that epigenetic alterations of HOX genes can be correlated with PCOS and consequently female infertility. This finding might offer additional definitions of PCOS, and eventually provides insight for novel treatments with epidrugs for this disease.

Keywords: epigenetic, HOX genes, PCOS, female infertility

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17 Epigenetic Modification Observed in Yeast Chromatin Remodeler Ino80p

Authors: Chang-Hui Shen, Michelle Esposito, Andrew J. Shen, Michael Adejokun, Diana Laterman

Abstract:

The packaging of DNA into nucleosomes is critical to genomic compaction, yet it can leave gene promoters inaccessible to activator proteins or transcription machinery and thus prevents transcriptional initiation. Both chromatin remodelers and histone acetylases (HATs) are the two main transcription co-activators that can reconfigure chromatin structure for transcriptional activation. Ino80p is the core component of the INO80 remodeling complex. Recently, it was shown that Ino80p dissociates from the yeast INO1 promoter after induction. However, when certain HATs were deleted or mutated, Ino80p accumulated at the promoters during gene activation. This suggests a link between HATs’ presence and Ino80p’s dissociation. However, it has yet to be demonstrated that Ino80p can be acetylated. To determine if Ino80p can be acetylated, wild-type Saccharomyces cerevisiae cells carrying Ino80p engineered with a double FLAG tag (MATa INO80-FLAG his3∆200 leu2∆0 met15∆0 trp1∆63 ura3∆0) were grown to mid log phase, as were non-tagged wild type (WT) (MATa his3∆200 leu2∆0 met15∆0 trp1∆63 ura3∆0) and ino80∆ (MATa ino80∆::TRP1 his3∆200 leu2∆0 met15∆0 trp1∆63 ura3∆0) cells as controls. Cells were harvested, and the cell lysates were subjected to immunoprecipitation (IP) with α-FLAG resin to isolate Ino80p. These eluted IP samples were subjected to SDS-PAGE and Western blot analysis. Subsequently, the blots were probed with the α-FLAG and α-acetyl lysine antibodies, respectively. For the blot probed with α-FLAG, one prominent band was shown in the INO80-FLAG cells, but no band was detected in the IP samples from the WT and ino80∆ cells. For the blot probed with the α-acetyl lysine antibody, we detected acetylated Ino80p in the INO80-FLAG strain while no bands were observed in the control strains. As such, our results showed that Ino80p can be acetylated. This acetylation can explain the co-activator’s recruitment patterns observed in current gene activation models. In yeast INO1, it has been shown that Ino80p is recruited to the promoter during repression, and then dissociates from the promoter once de-repression begins. Histone acetylases, on the other hand, have the opposite pattern of recruitment, as they have an increased presence at the promoter as INO1 de-repression commences. This Ino80p recruitment pattern significantly changes when HAT mutant strains are studied. It was observed that instead of dissociating, Ino80p accumulates at the promoter in the absence of functional HATs, such as Gcn5p or Esa1p, under de-repressing processes. As such, Ino80p acetylation may be required for its proper dissociation from the promoters. The remodelers’ dissociation mechanism may also have a wide range of implications with respect to transcriptional initiation, elongation, or even repression as it allows for increased spatial access to the promoter for the various transcription factors and regulators that need to bind in that region. Our findings here suggest a previously uncharacterized interaction between Ino80p and other co-activators recruited to promoters. As such, further analysis of Ino80p acetylation not only will provide insight into the role of epigenetic modifications in transcriptional activation, but also gives insight into the interactions occurring between co-activators at gene promoters during gene regulation.

Keywords: acetylation, chromatin remodeler, epigenetic modification, Ino80p

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16 Biostimulant Activity of Chitooligomers: Effect of Different Degrees of Acetylation and Polymerization on Wheat Seedlings under Salt Stress

Authors: Xiaoqian Zhang, Ping Zou, Pengcheng Li

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Salt stress is one of the most serious abiotic stresses, and it can lead to the reduction of agricultural productivity. High salt concentration makes it more difficult for roots to absorb water and disturbs the homeostasis of cellular ions resulting in osmotic stress, ion toxicity and generation of reactive oxygen species (ROS). Compared with the normal physiological conditions, salt stress could inhibit the photosynthesis, break metabolic balance and damage cellular structures, and ultimately results in the reduction of crop yield. Therefore it is vital to develop practical methods for improving the salt tolerance of plants. Chitooligomers (COS) is partially depolymerized products of chitosan, which is consisted of D-glucosamine and N-acetyl-D-glucosamine. In agriculture, COS has the ability to promote plant growth and induce plant innate immunity. The bioactivity of COS closely related to its degree of polymerization (DP) and acetylation (DA). However, most of the previous reports fail to mention the function of COS with different DP and DAs in improving the capacity of plants against salt stress. Accordingly, in this study, chitooligomers (COS) with different degrees of DAs were used to test wheat seedlings response to salt stress. In addition, the determined degrees of polymerization (DPs) COS(DP 4-12) and a heterogeneous COS mixture were applied to explore the relationship between the DP of COSs and its effect on the growth of wheat seedlings in response to salt stress. It showed that COSs, the exogenous elicitor, could promote the growth of wheat seedling, reduce the malondialdehyde (MDA) concentration, and increase the activities of antioxidant enzymes. The results of mRNA expression level test for salt stress-responsive genes indicated that COS keep plants away from being hurt by the salt stress via the regulation of the concentration and the increased antioxidant enzymes activities. Moreover, it was found that the activities of COS was closely related to its Das and COS (DA: 50%) displayed the best salt resistance activity to wheat seedlings. The results also showed that COS with different DP could promote the growth of wheat seedlings under salt stress. COS with a DP (6-8) showed better activities than the other tested samples, implied its activity had a close relationship with its DP. After treatment with chitohexaose, chitoheptaose, and chitooctaose, the photosynthetic parameters were improved obviously. The soluble sugar and proline contents were improved by 26.7%-53.3% and 43.6.0%-70.2%, respectively, while the concentration of malondialdehyde (MDA) was reduced by 36.8% - 49.6%. In addition, the antioxidant enzymes activities were clearly activated. At the molecular level, the results revealed that they could obviously induce the expression of Na+/H+ antiporter genes. In general, these results were fundamental to the study of action mechanism of COS on promoting plant growth under salt stress and the preparation of plant growth regulator.

Keywords: chitooligomers (COS), degree of polymerization (DP), degree of acetylation (DA), salt stress

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15 Linking Metabolism, Pluripotency and Epigenetic Changes during Early Differentiation of Embryonic Stem Cells

Authors: Arieh Moussaieff, Bénédicte Elena-Herrmann, Yaakov Nahmias, Daniel Aberdam

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Differentiation of pluripotent stem cells is a slow process, marked by the gradual loss of pluripotency factors over days in culture. While the first few days of differentiation show minor changes in the cellular transcriptome, intracellular signaling pathways remain largely unknown. Recently, several groups demonstrated that the metabolism of pluripotent mouse and human cells is different from that of somatic cells, showing a marked increase in glycolysis previously identified in cancer as the Warburg effect. Here, we sought to identify the earliest metabolic changes induced at the first hours of differentiation. High-resolution NMR analysis identified 35 metabolites and a distinct, gradual transition in metabolism during early differentiation. Metabolic and transcriptional analyses showed the induction of glycolysis toward acetate and acetyl-coA in pluripotent cells, and an increase in cholesterol biosynthesis during early differentiation. Importantly, this metabolic pathway regulated differentiation of human and mouse embryonic stem cells. Acetate delayed differentiation preventing differentiation-induced histone de-acetylation in a dose-dependent manner. Glycolytic inhibitors upstream of acetate caused differentiation of pluripotent cells, while those downstream delayed differentiation. Our data suggests that a rapid loss of glycolysis in early differentiation down-regulates acetate and acetyl-coA production, causing a loss of histone acetylation and concomitant loss of pluripotency. It demonstrate that pluripotent stem cells utilize a novel metabolism pathway to maintain pluripotency through acetate/acetyl-coA and highlights the important role metabolism plays in pluripotency and early differentiation of stem cells.

Keywords: pluripotency, metabolomics, epigenetics, acetyl-coA

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14 Chemical Modifications of Three Underutilized Vegetable Fibres for Improved Composite Value Addition and Dye Absorption Performance

Authors: Abayomi O. Adetuyi, Jamiu M. Jabar, Samuel O. Afolabi

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Vegetable fibres are classes of fibres of low density, biodegradable and non-abrasive that are largely abundant fibre materials with specific properties and mostly found/ obtained in plants on earth surface. They are classified into three categories, depending on the part of the plant from which they are gotten from namely: fruit, Blast and Leaf fibre. Ever since four/five millennium B.C, attention has been focussing on the commonest and highly utilized cotton fibre obtained from the fruit of cotton plants (Gossypium spp), for the production of cotton fabric used in every home today. The present study, therefore, focused on the ability of three underutilized vegetable (fruit) fibres namely: coir fiber (Eleas coniferus), palm kernel fiber and empty fruit bunch fiber (Elias guinensis) through chemical modifications for better composite value addition performance to polyurethane form and dye adsorption. These fibres were sourced from their parents’ plants, identified and cleansed with 2% hot detergent solution 1:100, rinsed in distilled water and oven-dried to constant weight, before been chemically modified through alkali bleaching, mercerization and acetylation. The alkali bleaching involves treating 0.5g of each fiber material with 100 mL of 2% H2O2 in 25 % NaOH solution with refluxing for 2 h. While that of mercerization and acetylation involves the use of 5% sodium hydroxide NaOH solution for 2 h and 10% acetic acid- acetic anhydride 1:1 (v/v) (CH3COOH) / (CH3CO)2O solution with conc. H2SO4 as catalyst for 1 h, respectively on the fibres. All were subsequently washed thoroughly with distilled water and oven dried at 105 0C for 1 h. These modified fibres were incorporated as composite into polyurethane form and used in dye adsorption study of indigo. The first two treatments led to fiber weight reduction, while the acidified acetic anhydride treatment gave the fibers weight increment. All the treated fibers were found to be of less hydrophilic nature, better mechanical properties, higher thermal stabilities as well as better adsorption surfaces/capacities than the untreated ones. These were confirmed by gravimetric analysis, Instron Universal Testing Machine, Thermogravimetric Analyser and the Scanning Electron Microscope (SEM) respectively. The fiber morphology of the modified fibers showed smoother surfaces than unmodified fibres.The empty fruit bunch fibre and the coconut coir fibre are better than the palm kernel fibres as reinforcers for composites or as adsorbents for waste-water treatment. Acetylation and alkaline bleaching treatment improve the potentials of the fibres more than mercerization treatment. Conclusively, vegetable fibres, especially empty fruit bunch fibre and the coconut coir fibre, which are cheap, abundant and underutilized, can replace the very costly powdered activated carbon in wastewater treatment and as reinforcer in foam.

Keywords: chemical modification, industrial application, value addition, vegetable fibre

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13 Analytical Tools for Multi-Residue Analysis of Some Oxygenated Metabolites of PAHs (Hydroxylated, Quinones) in Sediments

Authors: I. Berger, N. Machour, F. Portet-Koltalo

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Polycyclic aromatic hydrocarbons (PAHs) are toxic and carcinogenic pollutants produced in majority by incomplete combustion processes in industrialized and urbanized areas. After being emitted in atmosphere, these persistent contaminants are deposited to soils or sediments. Even if persistent, some can be partially degraded (photodegradation, biodegradation, chemical oxidation) and they lead to oxygenated metabolites (oxy-PAHs) which can be more toxic than their parent PAH. Oxy-PAHs are less measured than PAHs in sediments and this study aims to compare different analytical tools in order to extract and quantify a mixture of four hydroxylated PAHs (OH-PAHs) and four carbonyl PAHs (quinones) in sediments. Methodologies: Two analytical systems – HPLC with on-line UV and fluorescence detectors (HPLC-UV-FLD) and GC coupled to a mass spectrometer (GC-MS) – were compared to separate and quantify oxy-PAHs. Microwave assisted extraction (MAE) was optimized to extract oxy-PAHs from sediments. Results: First OH-PAHs and quinones were analyzed in HPLC with on-line UV and fluorimetric detectors. OH-PAHs were detected with the sensitive FLD, but the non-fluorescent quinones were detected with UV. The limits of detection (LOD)s obtained were in the range (2-3)×10-4 mg/L for OH-PAHs and (2-3)×10-3 mg/L for quinones. Second, even if GC-MS is not well adapted to the analysis of the thermodegradable OH-PAHs and quinones without any derivatization step, it was used because of the advantages of the detector in terms of identification and of GC in terms of efficiency. Without derivatization, only two of the four quinones were detected in the range 1-10 mg/L (LODs=0.3-1.2 mg/L) and LODs were neither very satisfying for the four OH-PAHs (0.18-0.6 mg/L). So two derivatization processes were optimized, comparing to literature: one for silylation of OH-PAHs, one for acetylation of quinones. Silylation using BSTFA/TCMS 99/1 was enhanced using a mixture of catalyst solvents (pyridine/ethyle acetate) and finding the appropriate reaction duration (5-60 minutes). Acetylation was optimized at different steps of the process, including the initial volume of compounds to derivatize, the added amounts of Zn (0.1-0.25 g), the nature of the derivatization product (acetic anhydride, heptafluorobutyric acid…) and the liquid/liquid extraction at the end of the process. After derivatization, LODs were decreased by a factor 3 for OH-PAHs and by a factor 4 for quinones, all the quinones being now detected. Thereafter, quinones and OH-PAHs were extracted from spiked sediments using microwave assisted extraction (MAE) followed by GC-MS analysis. Several mixtures of solvents of different volumes (10-25 mL) and using different extraction temperatures (80-120°C) were tested to obtain the best recovery yields. Satisfactory recoveries could be obtained for quinones (70-96%) and for OH-PAHs (70-104%). Temperature was a critical factor which had to be controlled to avoid oxy-PAHs degradation during the MAE extraction process. Conclusion: Even if MAE-GC-MS was satisfactory to analyze these oxy-PAHs, MAE optimization has to be carried on to obtain a most appropriate extraction solvent mixture, allowing a direct injection in the HPLC-UV-FLD system, which is more sensitive than GC-MS and does not necessitate a previous long derivatization step.

Keywords: derivatizations for GC-MS, microwave assisted extraction, on-line HPLC-UV-FLD, oxygenated PAHs, polluted sediments

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12 Sex Differences in Age-Related AMPK-Sirt1 Axis Alteration in Human Heart

Authors: Maria Luisa Barcena De Arellano, Sofya Pozdniakova, Pavelas Karkacas, Anja Kuhl, Istvan Baczko, Yury Ladilov, Vera Regitz-Zagrosek

Abstract:

Introduction: Aging is associated with deterioration of the physiological function, leading to systemic inflammation and mitochondrial dysfunction that promote the development of cardiovascular diseases. Sex differences in aging-related cardiovascular diseases have been postulated. However, their precise mechanisms remain unclear. In the current study, we aimed to investigate the sex difference in the age-related alteration in Sirt1-AMPK signaling and its relation to the mitochondrial biogenesis and inflammation. Methods: Male and female human non-disease lateral left ventricular wall tissue (young (17–40 years; n= 7 male and 7 female) and old (50–68 years; n= 9 male and 8 female)) were used. qRT-PCR, western blot and immunohistochemistry assays were performed for expression analyses of Sirt1, AMPK, pAMPK, ac-Ku70, TFAM, PGC-1α, Sirt3, SOD2 and catalase. CD68 was used as a marker for macrophages and the ratio of IL-12:IL10 (pro-inflammatory phenotype (high IL-12/low IL-10) and anti-inflammatory phenotype (low IL-12/high IL-10) was used to examine the inflammatory stage in the heart. Results: Sirt1 expression was significantly higher in young females compared to young males, whereas in aged hearts Sirt1 expression was significantly downregulated in females, but not in males. In line with the Sirt1 downregulation in aged females, acetylation of nuclear Ku70, a direct target of Sirt1, in aged female hearts was significantly elevated. The activity of AMPK was significantly decreased in aged individuals, however no sex differences in the AMPK expression or activity were found in young or old individuals. The expression of mitochondrial proteins TOM40, SOD2 and Sirt3 was significantly higher in young females compared to young males, while in aged female hearts SOD2 and TOM40 were downregulated. In addition, the expression of catalase, a key cytosolic and mitochondrial anti-oxidative enzyme was significantly higher in young females and this female sex benefit was lost in aged hearts. In addition, the number of cardiac macrophages was significantly increased in old female, but not in male hearts. Consistently, the pro-inflammatory shift in old females was further confirmed by differences in the IL12/IL10 ratio in young female cardiac tissue in a favour of the anti-inflammatory mediator IL-10 (ratio 1:4) compared to young males (ratio 1:1). The anti-inflammatory environment in the heart was lost in aged females (ratio 1:1). Conclusion: Aging leads to the significant downregulation of Sirt1 expression and elevated acetylation of Ku70 in female, but not in male hearts. Furthermore, a beneficial upregulation of mitochondrial and anti-oxidative proteins in young females is lost with aging. Moreover, the malfunctions in the expression of Sirt1 and mitochondrial proteins in aged female hearts is accompanied by a significant pro-inflammatory shift. The study provides a molecular basis for the increased incidence of cardiovascular diseases in old women.

Keywords: inflammation, mitochondrial dysfunction, aging, Sirt1-AMPK axis

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11 Facile Synthetic Process for Lamivudine and Emtricitabine

Authors: Devender Mandala, Paul Watts

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Cis-Nucleosides mainly lamivudine (3TC) and emtricitabine (FTC) are an important tool in the treatment of Human immune deficiency virus (HIV), Hepatitis B virus (HBV) and Human T-Lymotropoic virus (HTLV). Lamivudine and emtricitabine are potent nucleoside analog reverse transcriptase inhibitors (nRTI). These two drugs are synthesized by a four-stage process from the starting materials: menthyl glyoxylate hydrate and 1,4-dithane-2,5-diol to produce the 5-hydroxy oxathiolane which upon acetylation with acetic anhydride to yield 5-acetoxy oxathiolane. Then glycosylation of this acetyl product with silyl protected nucleoside to produce the intermediate. The reduction of this intermediates can provide the final targets. Although there are several different methods reported for the synthesis of lamivudine and emtricitabine as a single enantiomer, we required an efficient route, which was suitable for large-scale synthesis to support the development of these compounds. In this process, we successfully prepared the intermediates of lamivudine and emtricitabine without using any solvents and catalyst, thus promoting the green synthesis. All the synthesized compound were confirmed by TLC, GC, Mass, NMR and 13C NMR spectroscopy.

Keywords: emtricitabine, green synthesis, lamivudine, nucleoside

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10 Synthesis of Green Fuel Additive from Waste Bio-Glycerol

Authors: Ala’a H. Al-Muhtaseb, Farrukh Jamil, Lamya Al-Haj, Mohab Al-Hinai

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Bio-glycerol is considered as high boiling polar triol and immiscible with fossil fuel fractions due to which it is transformed into its respective ketals and acetals which help to improve the quality of diesel emitting less amount of aldehydes and carbon monoxide. Solketal visual appearance is transparent and it is odorless organic liquid used as fuel additive for diesel to improve its cold flow properties. Condensation of bio-glycerol with bio-acetone in presence of beta zeolite has been done for synthesizing solketal. It was observed that glycerol conversion and selectivity of solketal was largely effected by temperature, as it increases from 40 ºC to 60 ºC the conversion of glycerol rises from 80.04 % to 94.26 % and selectivity of solketal from 80.0 % to 94.21 % but further increase in temperature to 100 ºC glycerol conversion reduced to 93.06 % and solketal selectivity to 92.08 %. At the optimum conditions, the bio-glycerol conversion and solketal yield were about 94.26% and 94.21wt% respectively. This process offers an attractive route for converting bio-glycerol, the main by-product of biodiesel to solketal with bio-acetone; a value-added green product with potential industrial applications as a valuable green fuel additive or combustion promoter for gasoline/diesel engines.

Keywords: bio-acetone, bio-glycerol, acetylation, solketal

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9 Water Absorption Studies on Natural Fiber Reinforced Polymer Composites

Authors: G. L. Devnani, Shishir Sinha

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In the recent years, researchers have drawn their focus on natural fibers reinforced composite materials because of their excellent properties like low cost, lower weight, better tensile and flexural strengths, biodegradability etc. There is little concern however that when these materials are put in moist conditions for long duration, their mechanical properties degrade. Therefore, in order to take maximum advantage of these novel materials, one should have a complete understanding of their moisture or water absorption phenomena. Various fiber surface treatment methods like alkaline treatment, acetylation etc. have also been suggested for reduction in water absorption of these composites. In the present study, a detailed review is done for water absorption behavior of natural fiber reinforced polymer composites, and experiments also have been performed on these composites with varying the parameters like fiber loading etc. for understanding the water absorption kinetics. Various surface treatment methods also performed to reduce the water absorption behavior of these materials and effort is made to develop a proper understanding of water absorption mechanism mathematically and experimentally for full potential utilization of natural fiber reinforced polymer composite materials.

Keywords: alkaline treatment, composites, natural fiber, water absorption

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8 Autophagy Suppresses Tumorigenesis through Upregulation of MiR-449a in Colorectal Cancer

Authors: Sheng-Hui Lan, Shan-Ying Wu, Shu-Ching Lin, Wei-Chen Wang, Hsiao-Sheng Liu

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Autophagy is an essential mechanism to maintain cellular homeostasis through its degradation function, and the autophagy deficiency is related various diseases including tumorigenesis in several cancers. MicroRNAs (miRNAs) are small none coding RNAs, which regulate gene expression through degradation of mRNA or inhibition of translation. However, the relationship between autophagy deficiency and dysregulated miRNAs is still unclear. We revealed a mechanism that autophagy up-regulates miR-449a expression at the transcriptional level through activation of forkhead transcription factor family member FoxO1 and then suppresses tumorigenesis in CRC. Our data showed that the autophagic activity and miR-449a expression were lower in colorectal cancer (CRC) and has a positive correlation. We further reveal that autophagy degrades p300 expression and then suppresses acetylation of FoxO1. Under autophagic induction conditions, FoxO1 is transported from the cytoplasm to the nucleus and binds to the miR-449a promoter and then promotes miR-449a expression. In addition, either miR-449a overexpression or amiodarone-induced autophagy inhibits cell cycle progression, proliferation, colony formation migration, invasion, and tumor formation of SW480 cells. Our findings indicate that autophagy inducers may have the potential to be used for prevention and treatment of CRC through upregulation of miR-449a expression.

Keywords: autophagy, MiR-449a, FoxO1, colorectal cancer

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7 Preparation and Characterization of Bioplastic from Sorghum Husks

Authors: Hannatu Abubakar Sani, Abubakar Umar Birnin Yauri, Aliyu Muhammad, Mujahid Salau, Aminu Musa, Hadiza Adamu Kwazo

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The increase in the global population and advances in technology have made plastic materials to have wide applications in every aspect of life. However, the non-biodegradability of these petrochemical-based materials and their increasing accumulation in the environment has been a threat to the planet and has been a source of environmental concerns and hence, the driving force in the search for ‘green’ alternatives for which agricultural waste remains the front liner. Sorghum husk, an agricultural waste with potentials as a raw material in the production of bioplastic, was used in this research to prepare bioplastic using sulphuric acid-catalyzed acetylation process. The prepared bioplastic was characterized by X-ray diffraction and Fourier transform infrared spectroscopy (FTIR), and the structure of the prepared bioplastic was confirmed. The Fourier transform infrared spectroscopy (FTIR) spectra of the product displayed the presence of OH, C-H, C=O, and C-O absorption peaks. The bioplastic obtained is biodegradable and is affected by acid, salt, and alkali to a lesser extent. Other tests like solubility and swelling studies were carried out to ensure the commercial properties of these bioplastic materials. Therefore, this revealed that new bioplastics with better environmental and sustainable properties could be produced from agricultural waste, which may have applications in many industries.

Keywords: agricultural waste, bioplastic, characterization, Sorghum Husk

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6 Effects of Interfacial Modification Techniques on the Mechanical Properties of Natural Particle Based Polymer Composites

Authors: Bahar Basturk, Secil Celik Erbas, Sevket Can Sarikaya

Abstract:

Composites combining the particulates and polymer components have attracted great interest in various application areas such as packaging, furniture, electronics and automotive industries. For strengthening the plastic matrices, the utilization of natural fillers instead of traditional reinforcement materials has received increased attention. The properties of natural filler based polymer composites (NFPC) may be improved by applying proper surface modification techniques to the powder phase of the structures. In this study, acorn powder-epoxy and pine corn powder-epoxy composites containing up to 45% weight percent particulates were prepared by casting method. Alkali treatment and acetylation techniques were carried out to the natural particulates for investigating their influences under mechanical forces. The effects of filler type and content on the tensile properties of the composites were compared with neat epoxy. According to the quasi-static tensile tests, the pine cone based composites showed slightly higher rigidity and strength properties compared to the acorn reinforced samples. Furthermore, the structures independent of powder type and surface modification technique, showed higher tensile properties with increasing the particle content.

Keywords: natural fillers, polymer composites, surface modifications, tensile properties

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5 Methylation Analysis of PHF20L1 and DACT2 Gene Promoters in Women with Breast Cancer

Authors: Marta E. Hernandez-Caballero, Veronica Borgonio-Cuadra, Antonio Miranda-Duarte, Xochitl Rojas-Toledo, Normand Garcia-Hernandez, Maura Cardenas-Garcia, Teresa Abad-Camacho

Abstract:

Breast cancer (BC) is the most common tumor in women over the world. DNA methylation is an epigenetic modification critical in CpG sites, aberrant methylation of CpG islands in promoters is a hallmark of cancer. So, gene expression can be regulated by alterations in DNA methylation. In cell lines DACT2 gene reduces the growth and migration of tumor cells by its participation in the suppression of TGFb/SMAD2/3. PHF20L1 is involved in histone acetylation therefore, it regulates transcription. Our aim was to analyze the methylation status of the DACT2 and PHF20L1 promoter regions in tumoral and healthy mammary tissue from women with BC in different progression states. The study included 77 patients from Centro Medico Nacional La Raza in Mexico City. After identifying a CpG island in DACT2 and PHF20L1 promoters, DNA methylation status was analyzed through sodium bisulfite with subsequent amplification using methylation-specific PCR. Results revealed no changes in methylation status of PHF20L1 and cancer stages (II y III) or in comparison to healthy tissues, it was demethylated. DACT2 promoter methylation was no significant between tumoral stages (II, P = 0.37; III, P = 0.17) or with healthy tissue. Previous data reported DACT2 methylated in nasopharyngeal carcinoma but in this study promoter methylation was not observed. PHF20L1 protein contains N-terminal Tudor and C-terminal plant homeodomain domains, it has been suggested that can stabilize DNMT1 regulating DNA methylation, therefore, was associated with poor prognostic in BC. We found no evidence of methylation in patients and controls in PHF20L1 promoter, so its association with BC may have no direct relation with promoter methylation. More studies including other methylation sites in these genes in BC are necessary.

Keywords: bisulfite conversion, breast cancer, DACT2, DNA methylation, PHF20L1, tumoral status

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4 Lignin Pyrolysis to Value-Added Chemicals: A Mechanistic Approach

Authors: Binod Shrestha, Sandrine Hoppe, Thierry Ghislain, Phillipe Marchal, Nicolas Brosse, Anthony Dufour

Abstract:

The thermochemical conversion of lignin has received an increasing interest in the frame of different biorefinery concepts for the production of chemicals or energy. It is needed to better understand the physical and chemical conversion of lignin for feeder and reactor designs. In-situ rheology reveals the viscoelastic behaviour of lignin upon thermal conversion. The softening, re-solidification (char formation), swelling and shrinking behaviours are quantified during pyrolysis in real-time [1]. The in-situ rheology of an alkali lignin (Protobind 1000) was conducted in high torque controlled strain rheometer from 35°C to 400°C with a heating rate of 5°C.min-1. The swelling, through glass phase transition overlapped with depolymerization, and solidification (crosslinking and “char” formation) are two main phenomena observed during lignin pyrolysis. The onset of temperatures for softening and solidification for this lignin has been found to be 141°C and 248°C respectively. An ex-situ characterization of lignin/char residues obtained at different temperatures after quenching in the rheometer gives a clear understanding of the pathway of lignin degradation. The lignin residues were sampled from the mid-point temperatures of the softening range and solidification range to study the chemical transformations undergoing. Elemental analysis, FTIR and solid state NMR were conducted after quenching the solid residues (lignin/char). The quenched solid was also extracted by suitable solvent and followed by acetylation and GPC-UV analysis. The combination of 13C NMR and GPC-UV reveals the depolymerization followed by crosslinking of lignin/char. NMR and FTIR provide the evolution of functional moieties upon temperature. Physical and chemical mechanisms occurring during lignin pyrolysis are accounted in this study. Thanks to all these complementary methods.

Keywords: pyrolysis, bio-chemicals, valorization, mechanism, softening, solidification, cross linking, rheology, spectroscopic methods

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3 ICAM1 Expression is Enhanced by TNFa through Histone Methylation in Human Brain Microvessel Cells

Authors: Ji-Young Choi, Jungjin Kim, Sang-Sun Yun, Sangmee Ahn Jo

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Intracellular adhesion molecule1 (ICAM1) is a mediator of inflammation and involved in adhesion and transmigration of leukocytes to endothelial cells, resulting in enhancement of brain inflammation. We hypothesized that increase of ICAM1 expression in endothelial cells is an early step in the pathogenesis of brain diseases such as Alzheimer’s disease. Here, we report that ICAM1 expression is regulated by pro-inflammatory cytokine TNFa in human microvascular endothelial cell (HBMVEC). TNFa significantly increased ICAM1 mRNA and protein levels at the concentrations showing no cell toxicity. This increase was also shown in micro vessels of mouse brain 24 hours after treatment with TNFa (8 mg/kg, i.v). We then investigated the epigenetic mechanism involved in the induction of ICAM1 expression. Chromatin immunoprecipitation assay revealed that TNFa reduced methylation of histone3K9 (H3K9-2me) and histone3K27 (H3K27-3me), well-known modification as gene suppression, with in the ICAM1 promoter region. However, acetylation of H3K9 and H3K14, well-known modification as gene activation, was not changed by TNFa. Treatment of BIX01294, a specific inhibitor of histone methyltransferase G9a responsible for H3K9-2me, dramatically increased in ICAM1 mRNA and protein levels and overexpression of G9a gene suppressed TNFa-induced ICAM1 expression. In contrast, GSK126, an inhibitor of histone methyltransferase EZH2 responsible for H3K27-3me and valproic acid, an inhibitor of histone deacetylase (HDAC) did not affect ICAM1 expression. These results suggested that histone3 methylation is involved in ICAM1 repression. Moreover, TNFa or BIX01294-induced ICAM induction resulted in both enhancements in adhesion and transmigration of leukocyte on endothelial cell. This study demonstrates that TNFa upregulates ICAM1 expression through H3K9-2me and H3K27-3me within the ICAM1 promoter region, in which G9a is likely to play a pivotal role in ICAM1 transcription. Our study provides a novel mechanism for ICAM1 transcription regulation in HBMVEC.

Keywords: ICAM1, TNFa, HBMVEC, H3K9-2me

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2 Sorption Properties of Hemp Cellulosic Byproducts for Petroleum Spills and Water

Authors: M. Soleimani, D. Cree, C. Chafe, L. Bates

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The accidental release of petroleum products into the environment could have harmful consequences to our ecosystem. Different techniques such as mechanical separation, membrane filtration, incineration, treatment processes using enzymes and dispersants, bioremediation, and sorption process using sorbents have been applied for oil spill remediation. Most of the techniques investigated are too costly or do not have high enough efficiency. This study was conducted to determine the sorption performance of hemp byproducts (cellulosic materials) in terms of sorption capacity and kinetics for hydrophobic and hydrophilic fluids. In this study, heavy oil, light oil, diesel fuel, and water/water vapor were used as sorbate fluids. Hemp stalk in different forms, including loose material (hammer milled (HM) and shredded (Sh) with low bulk densities) and densified forms (pellet form (P) and crumbled pellets (CP)) with high bulk densities, were used as sorbents. The sorption/retention tests were conducted according to ASTM 726 standard. For a quick-purpose application of the sorbents, the sorption tests were conducted for 15 min, and for an ideal sorption capacity of the materials, the tests were carried out for 24 h. During the test, the sorbent material was exposed to the fluid by immersion, followed by filtration through a stainless-steel wire screen. Water vapor adsorption was carried out in a controlled environment chamber with the capability of controlling relative humidity (RH) and temperature. To determine the kinetics of sorption for each fluid and sorbent, the retention capacity also was determined intervalley for up to 24 h. To analyze the kinetics of sorption, pseudo-first-order, pseudo-second order and intraparticle diffusion models were employed with the objective of minimal deviation of the experimental results from the models. The results indicated that HM and Sh materials had the highest sorption capacity for the hydrophobic fluids with approximately 6 times compared to P and CP materials. For example, average retention values of heavy oil on HM and Sh was 560% and 470% of the mass of the sorbents, respectively. Whereas, the retention of heavy oil on P and CP was up to 85% of the mass of the sorbents. This lower sorption capacity for P and CP can be due to the less exposed surface area of these materials and compacted voids or capillary tubes in the structures. For water uptake application, HM and Sh resulted in at least 40% higher sorption capacity compared to those obtained for P and CP. On average, the performance of sorbate uptake from high to low was as follows: water, heavy oil, light oil, diesel fuel. The kinetic analysis indicated that the second-pseudo order model can describe the sorption process of the oil and diesel better than other models. However, the kinetics of water absorption was better described by the pseudo-first-order model. Acetylation of HM materials could improve its oil and diesel sorption to some extent. Water vapor adsorption of hemp fiber was a function of temperature and RH, and among the models studied, the modified Oswin model was the best model in describing this phenomenon.

Keywords: environment, fiber, petroleum, sorption

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1 Removal of Heavy Metals by Ultrafiltration Assisted with Chitosan or Carboxy-Methyl Cellulose

Authors: Boukary Lam, Sebastien Deon, Patrick Fievet, Nadia Crini, Gregorio Crini

Abstract:

Treatment of heavy metal-contaminated industrial wastewater has become a major challenge over the last decades. Conventional processes for the treatment of metal-containing effluents do not always simultaneously satisfy both legislative and economic criteria. In this context, coupling of processes can then be a promising alternative to the conventional approaches used by industry. The polymer-assisted ultrafiltration (PAUF) process is one of these coupling processes. Its principle is based on a sequence of steps with reaction (e.g., complexation) between metal ions and a polymer and a step involving the rejection of the formed species by means of a UF membrane. Unlike free ions, which can cross the UF membrane due to their small size, the polymer/ion species, the size of which is larger than pore size, are rejected. The PAUF process was deeply investigated herein in the case of removal of nickel ions by adding chitosan and carboxymethyl cellulose (CMC). Experiments were conducted with synthetic solutions containing 1 to 100 ppm of nickel ions with or without the presence of NaCl (0.05 to 0.2 M), and an industrial discharge water (containing several metal ions) with and without polymer. Chitosan with a molecular weight of 1.8×105 g mol⁻¹ and a degree of acetylation close to 15% was used. CMC with a degree of substitution of 0.7 and a molecular weight of 9×105 g mol⁻¹ was employed. Filtration experiments were performed under cross-flow conditions with a filtration cell equipped with a polyamide thin film composite flat-sheet membrane (3.5 kDa). Without the step of polymer addition, it was found that nickel rejection decreases from 80 to 0% with increasing metal ion concentration and salt concentration. This behavior agrees qualitatively with the Donnan exclusion principle: the increase in the electrolyte concentration screens the electrostatic interaction between ions and the membrane fixed the charge, which decreases their rejection. It was shown that addition of a sufficient amount of polymer (greater than 10⁻² M of monomer unit) can offset this decrease and allow good metal removal. However, the permeation flux was found to be somewhat reduced due to the increase in osmotic pressure and viscosity. It was also highlighted that the increase in pH (from 3 to 9) has a strong influence on removal performances: the higher pH value, the better removal performance. The two polymers have shown similar performance enhancement at natural pH. However, chitosan has proved more efficient in slightly basic conditions (above its pKa) whereas CMC has demonstrated very weak rejection performances when pH is below its pKa. In terms of metal rejection, chitosan is thus probably the better option for basic or strongly acid (pH < 4) conditions. Nevertheless, CMC should probably be preferred to chitosan in natural conditions (5 < pH < 8) since its impact on the permeation flux is less significant. Finally, ultrafiltration of an industrial discharge water has shown that the increase in metal ion rejection induced by the polymer addition is very low due to the competing phenomenon between the various ions present in the complex mixture.

Keywords: carboxymethyl cellulose, chitosan, heavy metals, nickel ion, polymer-assisted ultrafiltration

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