Startseite Critical Roles of the PI3K-Akt-mTOR Signaling Pathway in Apoptosis and Autophagy of Astrocytes Induced by Methamphetamine
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Critical Roles of the PI3K-Akt-mTOR Signaling Pathway in Apoptosis and Autophagy of Astrocytes Induced by Methamphetamine

  • Han-Qing Liu , Ya-Wen An , A-Zhen Hu , Ming-Hua Li , Jue-Lian Wu , Li Liu , Yu Shi EMAIL logo , Guang-Hui Cui EMAIL logo und Yun Chen EMAIL logo
Veröffentlicht/Copyright: 22. Februar 2019

Abstract

This study aimed to reveal potential roles of the phosphatidylinositol 3 kinase (PI3K)-protein kinase B (Akt)-mammalian target of rapamycin (mTOR) signaling pathway in apoptosis and autophagy of astrocytes induced by methamphetamine (METH). A Cell Counting Kit-8 (CCK-8) was used to determine the reduction in proliferation of U-118 MG cells induced by METH. Hoechst 33258 and flow cytometry were used to observe the astrocytes. Western blot analysis was performed to evaluate protein expression and phosphorylation levels. METH inhibited the proliferation of U-118 MG cells and induced apoptosis and autophagy. Western blot analysis showed that the ratio of LC3-II/I was increased, whereas the expression of Bcl-2 was decreased. The phosphorylation cascade of kinases in the PI3K-Akt-mTOR signaling pathway was significantly inhibited by METH exposure, as were proteins downstream of mTORC1, such as p70s6k, rps6, 4EBP1 and eIF4E. METH inhibited proliferation of U-118 MG cells and induced apoptosis and autophagy. The PI3K-Akt-mTOR signaling pathway likely plays a critical role in these effects.

1 Introduction

Drug abuse has increased significantly year by year, both in China and the rest of the world, and has now reached epidemic proportions. Synthetic drugs, including methamphetamine (METH), have become the most widely abused substances, with their use far exceeding that of opiates. METH abusers are increasingly recognized to suffer from sustained brain damage, leading to cognitive and motor deficits. Although the mechanisms contributing to this damage are not fully understood, neuronal cell death has been observed in many animal models following METH exposure. This cell death, which involves cells in reward circuits such as the hippocampus, nucleus accumbens and corpus striatum, may contribute to the pathogenesis of neurotoxicity [1, 2, 3, 4].

Recently, METH-induced microglial activation and death have also been well documented [5, 6], although the relative roles of signaling pathways in apoptosis and autophagy remain to be elucidated. The phosphatidylinositol 3 kinase (PI3K)-protein kinase B (Akt)-mammalian target of rapamycin (mTOR) signaling pathway regulates numerous biological processes, including cell survival, proliferation and growth, and glucose metabolism [7, 8, 9], and has been demonstrated to be involved in the neuronal adaptations that underlie drug-related behaviors such as reward seeking and excessive drug intake [10]. It also has been reported that the PI3K-Akt-mTOR signaling pathway is the critical factor for cue-induced reinstatement of drug seeking [11]. In a previous study, we found that METH could induce conditioned place preference (CPP) in rats, and that this effect was regulated by the PI3K-AKT-mTOR signaling pathway [13]. However, the roles of PI3K-Akt-mTOR signaling pathway in regulating the survival and death of glial cells induced by METH were still unclear. Therefore, in the present study, we aimed to observe the inhibition of cell proliferation, apoptosis and autophagy of astrocytes induced by METH, and to reveal the role of the PI3K-Akt-mTOR signaling pathway. These studies might help us to understand the mechanisms of METH addiction and might provide some avenues for withdrawal treatment.

2 Materials and methods

2.1 Cell culture and treatment

METH (purity > 99%) was provided by the compulsory isolation and detoxification program operated in Shenzhen, China. U-118 MG cells were provided by the Hong Kong University of Science and Technology and cultured in DMEM supplemented with 10% FBS. All primary antibodies were diluted 1000 times with a primary antibody dilution buffer. Information regarding the reagents are summarized in Table. 1. Cells were added to well-plates at a density of 1 × 106 cells/mL and incubated at 37°C under an atmosphere containing 5% CO2. The concentrations of METH tested using the CCK-8 were 198, 296, 444, 667 and 1000 μg/mL. Six replicate wells were set for each concentration, and identical cells without METH treatment were used as a control. A solution of METH in DMEM (1000 μg/mL) was progressively diluted after sterilization by filtration. Cells were exposed to METH for 24 h and CCK-8 reagent (10 μL) was then added to the medium (100 μL). The cells were cultured at 37°C for 1 h and the absorbance of the cells at 450 nm, with a reference wavelength of 405 nm, was then determined using a Spectra Max i3x Microplate Reader (Molecular Devices, Shanghai, China). The inhibition of cell proliferation at every concentration (IC) was calculated with the formula IC (%) = (A0-AM)/A0×100, wherein A0 means the absorbance of the control group and AM represents the absorbance of groups treated with METH of different concentrations. A concentration-inhibition curve was fitted using the IC (%) as the Y-axis and concentrations of METH as the X-axis.

Table 1

Information of the reagents.

ReagentsAbbreviationPurity /cat.noSource
Cell Counting Kit-8CCK-8cat. no. CK04Dojindo, Tokyo, Japan
IP Lysis Buffer/cat. no. 87788Thermo Fisher Scientific, Waltham, MA, USA
Alexa FluorTM 488 Annexin V Kit/cat. no. V13241
phenyl methane sulfonyl fluoridePMSF≥98.5, HPLC
Hoechst 33258/≥98, HPLCBeyotime, Shanghai, China
Na3VO4/99.98, HPLCSigma-Aldrich, Burlington, MA, USA
NaF/≥99, HPLC
phospho-phosphatidylinositol 3 kinasep-PI3Kcat. no. PA5-17387Invitrogen, Carlsbad, CA, USA
phospho-p70 ribosomal s6 kinasep-p70s6kcat. no. 701083
phospho-ribosomal protein s6p-rps6cat. no. MA5-15140
phospho-4E binding protein 1p-4EBP1cat. no. PA5-17728
phospho-protein synthesis initiation factor 4Ep-eIF4Ecat. no. PA5-17919
BCL2-Associated XBaxcat. no. #2772Cell Signaling Technology, Danvers, MA, USA
B-cell lymphoma-2Bcl-2cat. no. #2782
Light Chain 3LC 3cat. No. #4108
β-actin/cat. no. #4970
phospho-protein kinase Bp-Aktcat. no. #4060
phospho-mammalian target of rapamycinp-mTORcat. no. #5536
HRP-labeled Goat Anti-Rabbit IgG/cat. no. A0208,Beyotime, Shanghai, China
primary antibody dilution buffe/cat. no. T143195Biosharp, Hefei, China
chemiluminescent HRP substrate/cat. no. WBKLS0100Millipore, MA, USA

An Alexa FluorTM 488 Annexin V/Dead Cell Apoptosis Kit, together with flow cytometry (FCW), was used to detect the presence of apoptotic cells. The concentrations of METH were 198, 296, 444, 667 and 1000 μg/mL, and identical cells without METH treatment were used as controls. After exposure to METH for 24 h, the cells were collected and resuspended at a concentration of 1 × 106 cells/mL. Annexin V solution (5 μL) and Alexa FluorTM 488 solution (5 μL) were added sequentially to the cell suspension (1 mL), and the mixture was then incubated in the dark at room temperature (RT) for 15 min. An Accuri® C6 flow cytometer (BD Pharmingen) was then used to identify apoptotic cells. For negative control, only Annexin V solution (5 μL) or Alexa FluorTM 488 solution (5 μL) was added.

Hoechst 33258 was dissolved in water (1 mg/mL) to provide a stock solution, which was diluted to 0.5 μg/mL for DNA staining. The cells were collected and fixed with ethyl acetate after exposure to METH for 24 h. The cells were then counterstained with Hoechst 33258 for 15 min at RT and observed using an IX51 fluorescence microscope (Olympus, Tokyo, Japan). Hoechst 33258 is a specific fluorescent DNA probe and the nuclei showed bright blue fluorescence after staining.

2.2 Western blot analysis

Cell culture and treatment were performed as described above, using METH concentrations of 198, 296, 444, 667 and 1000 μg/mL, and identical cells without METH treatment were used as control. After treatment with METH for 24 h, the cells were collected, placed in 1.5-mL microtubes and treated with cell lysis buffer containing PMSF (1 mM), Na3VO4 (1 mM) and NaF (20 mM). After 30 min at 4°C, the mixture of cells and lysis buffer was centrifuged at 12,000 g for 15 min at 4°C to separate the proteins. Loading buffer (5 ×) was added to the protein solutions at a ratio of 1: 4 and the samples were digested for 10 min at 98°C. Protein samples (5 μL) were loaded into the wells of polyacrylamide gels and separated by electrophoresis at 80–120 V. Proteins were transferred to polyvinylidene fluoride membranes with an electrical current of 0.33 A for 1.5 h. The membranes were incubated overnight at 4°C with primary antibodies and then for 1 h at RT with a horseradish peroxidase (HRP)-labeled Goat Anti-Rabbit IgG (1: 2000). Protein expression and phosphorylation levels were determined using a 5220S chemiluminescence detector (Tanon, Guangzhou, Guangdong, China) with immobilonTM Western chemiluminescent HRP substrate.

2.3 Statistical analysis

Values for the target protein levels were normalized to β-actin and analyzed using Quantity One software (version 4.4.0; Bio-Rad, Hercules, CA, USA). Data were expressed as means ± standard deviation (SD) and intergroup differences were compared by one-way ANOVA analysis, using SPSS 17.0 software (SPSS, Chicago, IL, USA). A probability (p) value of < 0.05 was considered to be statistically significant.

Ethical approval: The conducted research is not related to either human or animal use.

3 Results

3.1 Inhibition of astrocyte proliferation and induction of apoptosis by METH

The CCK-8 test, Hoechst 33258 staining, and FCW were used to observe METH-induced inhibition of proliferation and induction of apoptosis in U-118 MG cells. METH inhibited cell proliferation in a dose-dependent manner, with a positive logarithmic correlation between the concentration and the degree of inhibition (Figure 1). Obvious apoptotic features, such as nuclear retraction, were observed after Hoechst 33258 fluorescence staining (Figure 2). The FCW test, using Alexa FluorTM 488 Annexin V, indicated that METH dose-dependently induced cell apoptosis (Figure 3).

Figure 1 METH exerts dose-dependent inhibition of the proliferation of U-118 MG cells. The concentrations of METH were 198, 296, 444, 667 and 1000 μg/mL, and a CCK-8 test was used to evaluate the cell proliferation. Data are presented as means ± SD (n = 6), and the annotation ** indicates a p value < 0.01 versus control group.
Figure 1

METH exerts dose-dependent inhibition of the proliferation of U-118 MG cells. The concentrations of METH were 198, 296, 444, 667 and 1000 μg/mL, and a CCK-8 test was used to evaluate the cell proliferation. Data are presented as means ± SD (n = 6), and the annotation ** indicates a p value < 0.01 versus control group.

Figure 2 Immunocytochemical detection of METH induced apoptosis in U-118 MG cells. Hoechst 33258 staining indicated that METH induced U-118 MG cells apoptosis. Morphologic changes in nuclei were observed with Hoechst 33258 staining under fluorescence microscopy. (a) Control group. The cells were shaped normally and had round, intact nuclei; (b) METH 198 μg/mL group; (c) METH 296 μg/mL group; (d) METH 444 μg/mL group; (e) METH 667 μg/mL group group; (f) METH 1000 μg/mL group. Original magnification is 200×.
Figure 2

Immunocytochemical detection of METH induced apoptosis in U-118 MG cells. Hoechst 33258 staining indicated that METH induced U-118 MG cells apoptosis. Morphologic changes in nuclei were observed with Hoechst 33258 staining under fluorescence microscopy. (a) Control group. The cells were shaped normally and had round, intact nuclei; (b) METH 198 μg/mL group; (c) METH 296 μg/mL group; (d) METH 444 μg/mL group; (e) METH 667 μg/mL group group; (f) METH 1000 μg/mL group. Original magnification is 200×.

Figure 3 Cells were stained with Annexin V- Alexa FluorTM 488 for verifying the apoptotic or necrotic cell ratio. METH induced U-118 MG cells apoptosis. (a) Alexa FluorTM 488 positive control; (b) Annexin V positive control; (c) double staining positive control. (1) Natural group; (2) METH 198 μg/mL group; (3) METH 296 μg/mL group; (4) METH 444 μg/mL group; (5) METH 667 μg/mL group group; (6) METH 1000 μg/mL group.
Figure 3

Cells were stained with Annexin V- Alexa FluorTM 488 for verifying the apoptotic or necrotic cell ratio. METH induced U-118 MG cells apoptosis. (a) Alexa FluorTM 488 positive control; (b) Annexin V positive control; (c) double staining positive control. (1) Natural group; (2) METH 198 μg/mL group; (3) METH 296 μg/mL group; (4) METH 444 μg/mL group; (5) METH 667 μg/mL group group; (6) METH 1000 μg/mL group.

3.2 Changes of protein expression and phosphorylation levels after exposure to METH

Bcl-2, an inner mitochondrial membrane protein, inhibits apoptotic neuronal cell death. METH inhibited the expression of Bcl-2; however, the expression of Bax showed no significant differences (Figure 4a) Autophagy, a type of programmed cell death that is independent of apoptosis, is negatively regulated by the mTORC1 signaling pathway. The LC3-II/I ratio is generally considered to be an indicator of the autophagosome, which is necessary for autophagy. Here, the LC3-II/I ratio was significantly increased after

Figure 4 METH induced apoptosis and autophagy of U-118 MG cells. (a) the expression of Bcl-2 and Bax; (b) the expression of LC3. The annotation * indicates a p value < 0.05 versus control group. The annotation ** indicates a p value < 0.01 versus control group.
Figure 4

METH induced apoptosis and autophagy of U-118 MG cells. (a) the expression of Bcl-2 and Bax; (b) the expression of LC3. The annotation * indicates a p value < 0.05 versus control group. The annotation ** indicates a p value < 0.01 versus control group.

exposure to METH (Figure 4b) These results indicated that METH induced both apoptosis and autophagy of U-118 MG cells.

To reveal the role of the PI3K-Akt-mTOR signaling pathway, we measured the phosphorylation levels of these kinases after METH treatment. The results showed that METH correlated with dephosphorylation of PI3K (Figure 5a) Akt (Figure 5b) and mTOR (Figure 5c) where PI3K and mTOR are kinases upstream and downstream of Akt, respectively. It is suggested that METH inhibited the activation of this signaling pathway.

Figure 5 METH inhibited the activity of PI3K-Akt-mTOR signaling pathway. (a) the expression of p-PI3K; (b) the expression of p-Akt; (3) the expression of p-mTOR. The annotation * indicates a p value < 0.05 versus control group. The annotation ** indicates a p value < 0.01 versus control group.
Figure 5

METH inhibited the activity of PI3K-Akt-mTOR signaling pathway. (a) the expression of p-PI3K; (b) the expression of p-Akt; (3) the expression of p-mTOR. The annotation * indicates a p value < 0.05 versus control group. The annotation ** indicates a p value < 0.01 versus control group.

Further, we observed the changes of kinases downstream of the mTOR complex-1 (mTORC1), including p70s6k, rps6, 4EBP1 and eIF4E. Here, the phosphorylation cascades were found to be inhibited after exposure to METH (Figure 6 a,b,c,d)

Figure 6 METH inhibited phosphorylation of downstream kinases of mTORC1. (a) the expression of p-p70s6k; (b) the expression of p-rps6; (3) the expression of p-4EBP; (3) the expression of p-eIF4E. The annotation * indicates a p value < 0.05 versus control group. The annotation ** indicates a p value < 0.01 versus control group.
Figure 6

METH inhibited phosphorylation of downstream kinases of mTORC1. (a) the expression of p-p70s6k; (b) the expression of p-rps6; (3) the expression of p-4EBP; (3) the expression of p-eIF4E. The annotation * indicates a p value < 0.05 versus control group. The annotation ** indicates a p value < 0.01 versus control group.

4 Discussion

METH, a commonly abused illegal psychostimulant, has been shown to induce neurotoxicity via multiple mechanisms, likely involving reactive oxygen species (ROS), leading to cell death through the apoptotic pathway, disturbances in mitochondrial function, neuroinflammation and autophagy [13, 14, 15]. Microglial activation and death have been well documented to involve in neuroplasticity and drug addiction. However, effects of METH on gliacytes and the roles of the PI3K-Akt-mTOR signaling pathway are still unclear.

In this research, we found that METH dose-dependently inhibited cell proliferation of the glial cell line U-118 MG, and induced its apoptosis and autophagy. The Bcl-2 family is one of the foremost oncogene families involved in cell apoptosis. Expression of Bcl-2 inhibits cell death by decreasing cellular generation of ROS. Our results showed that treatment of U-118 MG cells with METH reduced the expression of Bcl-2. Previous studies have shown that METH inhibits the expression of Bcl-2 in various cells (SH-SY5Y, PC12, etc.), which is consistent with our results [16, 17, 18, 19]. These results provide potential mechanisms and help to explain METH-induced cell apoptosis.

The PI3K-Akt-mTOR signaling pathway has been demonstrated to be involved in addiction and cue-induced relapse related to several substances. In addition, this pathway is well known to regulate biological processes, including cell survival, proliferation, apoptosis and autophagy. Wu et al. [20] found that exposure to METH decreased phosphorylation within the Akt-GSK3β-mTOR pathway in PC12 cells. Lithium protected against METH-induced neurotoxicity by reversing the dephosphorylation of Akt-GSK3β-mTOR, and LY294002, a PI3K-Akt inhibitor, was found to suppress the protective effects of lithium. Lv et al. [21] found that thioredoxin-1, an important intra- and extra-cellular regulator of redox reactions, was induced by METH in PC12 cells and that the PI3K-Akt pathway was involved in the METH-induced increase of thioredoxin-1 expression.

These studies indicate that a variety of different mechanisms link the PI3K-Akt-mTOR pathway to METH-induced cell apoptosis. Our results show that exposure to METH also causes changes in phosphorylation of PI3K-Akt-mTOR in U-118 MG cells, which may be associated with the cell apoptosis induced by METH in these cells.

In terms of regulating protein synthesis, the downstream effectors of mTORC1 are mainly p70s6k, rps6, 4EBP and eIF4E. Autophagy might mediate neuroprotective effects following some types of brain damage, including METH intoxication. However, autophagy precipitates a peculiar form of cell death that contributes to some types of brain damage in other circumstances [22, 23, 24]. What is clear about autophagy for now is that it is regulated by the mTOR signaling pathway. However, the potential contribution of autophagy to METH-induced damage of neuronal cells remains unclear.

There is abundant evidence that mTORC1 and its downstream kinases participate in METH-induced glial autophagy and apoptosis. For example, Li et al. [25] found that METH elevated expression of LC3-II, a protein associated with the autophagosome, in PC 12 cells, but significantly inhibited p-mTOR. Taurine attenuated METH-induced autophagy and apoptosis via the mTOR pathway. Hebert & O’Callaghan [26] demonstrated that METH-induced glial activation in female C57BL/6J mice was associated with significant changes in activated forms of p70s6k. Using the SK-N-SH dopaminergic cell line, Kongsuphol et al. [27] investigated the effect of METH on autophagy and its upstream regulator, the mTOR signaling pathway, and found that METH induced expression of LC3-II but inhibited phosphorylation of mTOR and 4EBP1. Huang et al. [22] confirmed that METH induces apoptosis and autophagy, and that the protective effects of L-ascorbate are mediated by inhibiting production of ROS. Ma et al. [28] reported that chronic exposure to METH disrupted cells whereas acute exposure induced early pro-survival responses mediated through inactivation of the Akt-mTOR-p70s6k pathway.

In the present study, we found that METH decreased phosphorylation of mTORC1 and its downstream kinases, but increased the LC3-II/I ratio. These results indicate that inactivation of mTORC1 and activation of autophagy following dephosphorylation of PI3K-Akt might contribute to the apoptosis of U-118 MG cells.

The effects of METH on cell proliferation or apoptosis was dose-dependently. Interestingly, the activation or inhibition of METH on protein expression and phosphorylation does not follow a strict dose-dependence as reversions were observed at the concentration of 667 μg/mL for several proteins, although the overall trend was consistent. The complex nature of the dose-response relationship suggests that there are potential mechanisms other than the PI3K-Akt-mTOR signaling pathway involved in the apoptosis and autophagy induced by METH, and these alternative mechanisms deserve further discussion. Further studies should provide more direct evidence of connections, including demonstrations of the opposing effects of PI3K-Akt-mTOR agonists and inhibitors on apoptosis and direct observations of the activation of autophagy. In addition, protective effects of lithium and N-Acetyl-L-cysteine (NAC), which was proved to influence the PI3K-Akt-mTOR signaling pathway, might be observed in further studies. Such further studies might provide a basis for the treatment of METH induced addiction and nerve injuries.

In conclusion, our results indicated that METH inhibited proliferation of U-118 MG cells and induced apoptosis and autophagy in this cell line and that the PI3K-Akt-mTOR signaling pathway might play critical roles in these effects.

Acknowledgements

We thank the Shenzhen Science and Technology Planning Project (JCYJ20160427185055877, JCYJ20160429090753103, JCYJ20170412171856582), the Health and Family Planning Commission of Shenzhen (201605018), Shenzhen “Sanming” project (SZSM201512026), Guangdong Bureau of Traditional Chinese Medicine Project (20171228) and Guangdong Medical Research Project (B2018090) for financial support and the Compulsory Isolation and Detoxification Program of Shenzhen for kindly providing the METH.

  1. Disclosure Statement The authors declare no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

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Received: 2018-06-18
Accepted: 2018-12-06
Published Online: 2019-02-22

© 2019 Han-Qing Liu et al., published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 Public License.

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  10. Thermodynamic properties of the bubble growth process in a pool boiling of water-ethanol mixture two-component system
  11. Critical Roles of the PI3K-Akt-mTOR Signaling Pathway in Apoptosis and Autophagy of Astrocytes Induced by Methamphetamine
  12. Characteristics of Stable Hydrogen and Oxygen Isotopes of Soil Moisture under Different Land Use in Dry Hot Valley of Yuanmou
  13. Specific, highly sensitive and simple spectrofluorimetric method for quantification of daclatasvir in HCV human plasma patients and in tablets dosage form
  14. Chromium-modified cobalt molybdenum nitrides as catalysts for ammonia synthesis
  15. Langerhans cell-like dendritic cells treated with ginsenoside Rh2 regulate the differentiation of Th1 and Th2 cells in vivo
  16. Identification of Powdery Mildew Blumeria graminis f. sp. tritici Resistance Genes in Selected Wheat Varieties and Development of Multiplex PCR
  17. Computational Analysis of new Degree-based descriptors of oxide networks
  18. The Use Of Chemical Composition And Additives To Classify Petrol And Diesel Using Gas Chromatography–Mass Spectrometry And Chemometric Analysis: A Uk Study
  19. Minimal Energy Tree with 4 Branched Vertices
  20. Jatropha seed oil derived poly(esteramide-urethane)/ fumed silica nanocomposite coatings for corrosion protection
  21. Calculating topological indices of certain OTIS interconnection networks
  22. Energy storage analysis of R125 in UIO-66 and MOF-5 nanoparticles: A molecular simulation study
  23. Velvet Antler compounds targeting major cell signaling pathways in osteosarcoma - a new insight into mediating the process of invasion and metastasis in OS
  24. Effects of Azadirachta Indica Leaf Extract, Capping Agents, on the Synthesis of Pure And Cu Doped ZnO-Nanoparticles: A Green Approach and Microbial Activity
  25. Aqueous Micro-hydration of Na+(H2O)n=1-7 Clusters: DFT Study
  26. A proposed image-based detection of methamidophos pesticide using peroxyoxalate chemiluminescence system
  27. Phytochemical screening and estrogenic activity of total glycosides of Cistanche deserticola
  28. Biological evaluation of a series of benzothiazole derivatives as mosquitocidal agents
  29. Chemical pretreatments of Trapa bispinosa's peel (TBP) biosorbent to enhance adsorption capacity for Pb(ll)
  30. Dynamic Changes in MMP1 and TIMP1 in the Antifibrotic Process of Dahuang Zhechong Pill in Rats with Liver Fibrosis
  31. The Optimization and Production of Ginkgolide B Lipid Microemulsion
  32. Photodynamic Therapy Enhanced the Antitumor Effects of Berberine on HeLa Cells
  33. Chiral and Achiral Enantiomeric Separation of (±)-Alprenolol
  34. Correlation of Water Fluoride with Body Fluids, Dental Fluorosis and FT4, FT3 –TSH Disruption among Children in an Endemic Fluorosis area in Pakistan
  35. A one-step incubation ELISA kit for rapid determination of dibutyl phthalate in water, beverage and liquor
  36. Free Radical Scavenging Activity of Essential Oil of Eugenia caryophylata from Amboina Island and Derivatives of Eugenol
  37. Effects of Blue and Red Light On Growth And Nitrate Metabolism In Pakchoi
  38. miRNA-199a-5p functions as a tumor suppressor in prolactinomas
  39. Solar photodegradation of carbamazepine from aqueous solutions using a compound parabolic concentrator equipped with a sun tracking system
  40. Influence of sub-inhibitory concentration of selected plant essential oils on the physical and biochemical properties of Pseudomonas orientalis
  41. Preparation and spectroscopic studies of Fe(II), Ru(II), Pd(II) and Zn(II) complexes of Schiff base containing terephthalaldehyde and their transfer hydrogenation and Suzuki-Miyaura coupling reaction
  42. Complex formation in a liquid-liquid extraction-chromogenic system for vanadium(IV)
  43. Synthesis, characterization (IR, 1H, 13C & 31P NMR), fungicidal, herbicidal and molecular docking evaluation of steroid phosphorus compounds
  44. Analysis and Biological Evaluation of Arisaema Amuremse Maxim Essential Oil
  45. A preliminary assessment of potential ecological risk and soil contamination by heavy metals around a cement factory, western Saudi Arabia
  46. Anti- inflammatory effect of Prunus tomentosa Thunb total flavones in LPS-induced RAW264.7 cells
  47. Collaborative Influence of Elevated CO2 Concentration and High Temperature on Potato Biomass Accumulation and Characteristics
  48. Methods of extraction, physicochemical properties of alginates and their applications in biomedical field – a review
  49. Characteristics of liposomes derived from egg yolk
  50. Preparation of ternary ZnO/Ag/cellulose and its enhanced photocatalytic degradation property on phenol and benzene in VOCs
  51. Influence of Human Serum Albumin Glycation on the Binding Affinities for Natural Flavonoids
  52. Synthesis and antioxidant activity of 2-methylthio-pyrido[3,2-e][1,2,4] triazolo[1,5-a]pyrimidines
  53. Comparative study on the antioxidant activities of ten common flower teas from China
  54. Molecular Properties of Symmetrical Networks Using Topological Polynomials
  55. Synthesis of Co3O4 Nano Aggregates by Co-precipitation Method and its Catalytic and Fuel Additive Applications
  56. Phytochemical analysis, Antioxidant and Antiprotoscolices potential of ethanol extracts of selected plants species against Echinococcus granulosus: In-vitro study
  57. Silver nanoparticles enhanced fluorescence for sensitive determination of fluoroquinolones in water solutions
  58. Simultaneous Quantification of the New Psychoactive Substances 3-FMC, 3-FPM, 4-CEC, and 4-BMC in Human Blood using GC-MS
  59. Biodiesel Production by Lipids From Indonesian strain of Microalgae Chlorella vulgaris
  60. Miscibility studies of polystyrene/polyvinyl chloride blend in presence of organoclay
  61. Antibacterial Activities of Transition Metal complexes of Mesocyclic Amidine 1,4-diazacycloheptane (DACH)
  62. Novel 1,8-Naphthyridine Derivatives: Design, Synthesis and in vitro screening of their cytotoxic activity against MCF7 cell line
  63. Investigation of Stress Corrosion Cracking Behaviour of Mg-Al-Zn Alloys in Different pH Environments by SSRT Method
  64. Various Combinations of Flame Retardants for Poly (vinyl chloride)
  65. Phenolic compounds and biological activities of rye (Secale cereale L.) grains
  66. Oxidative degradation of gentamicin present in water by an electro-Fenton process and biodegradability improvement
  67. Optimizing Suitable Conditions for the Removal of Ammonium Nitrogen by a Microbe Isolated from Chicken Manure
  68. Anti-inflammatory, antipyretic, analgesic, and antioxidant activities of Haloxylon salicornicum aqueous fraction
  69. The anti-corrosion behaviour of Satureja montana L. extract on iron in NaCl solution
  70. Interleukin-4, hemopexin, and lipoprotein-associated phospholipase A2 are significantly increased in patients with unstable carotid plaque
  71. A comparative study of the crystal structures of 2-(4-(2-(4-(3-chlorophenyl)pipera -zinyl)ethyl) benzyl)isoindoline-1,3-dione by synchrotron radiation X-ray powder diffraction and single-crystal X-ray diffraction
  72. Conceptual DFT as a Novel Chemoinformatics Tool for Studying the Chemical Reactivity Properties of the Amatoxin Family of Fungal Peptides
  73. Occurrence of Aflatoxin M1 in Milk-based Mithae samples from Pakistan
  74. Kinetics of Iron Removal From Ti-Extraction Blast Furnace Slag by Chlorination Calcination
  75. Increasing the activity of DNAzyme based on the telomeric sequence: 2’-OMe-RNA and LNA modifications
  76. Exploring the optoelectronic properties of a chromene-appended pyrimidone derivative for photovoltaic applications
  77. Effect of He Qi San on DNA Methylation in Type 2 Diabetes Mellitus Patients with Phlegm-blood Stasis Syndrome
  78. Cyclodextrin potentiometric sensors based on selective recognition sites for procainamide: Comparative and theoretical study
  79. Greener synthesis of dimethyl carbonate from carbon dioxide and methanol using a tunable ionic liquid catalyst
  80. Nonisothermal Cold Crystallization Kinetics of Poly(lactic acid)/Bacterial Poly(hydroxyoctanoate) (PHO)/Talc
  81. Enhanced adsorption of sulfonamide antibiotics in water by modified biochar derived from bagasse
  82. Study on the Mechanism of Shugan Xiaozhi Fang on Cells with Non-alcoholic Fatty Liver Disease
  83. Comparative Effects of Salt and Alkali Stress on Antioxidant System in Cotton (Gossypium Hirsutum L.) Leaves
  84. Optimization of chromatographic systems for analysis of selected psychotropic drugs and their metabolites in serum and saliva by HPLC in order to monitor therapeutic drugs
  85. Electrocatalytic Properties of Ni-Doped BaFe12O19 for Oxygen Evolution in Alkaline Solution
  86. Study on the removal of high contents of ammonium from piggery wastewater by clinoptilolite and the corresponding mechanisms
  87. Phytochemistry and toxicological assessment of Bryonia dioica roots used in north-African alternative medicine
  88. The essential oil composition of selected Hemerocallis cultivars and their biological activity
  89. Mechanical Properties of Carbon Fiber Reinforced Nanocrystalline Nickel Composite Electroforming Deposit
  90. Anti-c-myc efficacy block EGFL7 induced prolactinoma tumorigenesis
  91. Topical Issue on Applications of Mathematics in Chemistry
  92. Zagreb Connection Number Index of Nanotubes and Regular Hexagonal Lattice
  93. The Sanskruti index of trees and unicyclic graphs
  94. Valency-based molecular descriptors of Bakelite network BNmn
  95. Computing Topological Indices for Para-Line Graphs of Anthracene
  96. Zagreb Polynomials and redefined Zagreb indices of Dendrimers and Polyomino Chains
  97. Topological Descriptor of 2-Dimensional Silicon Carbons and Their Applications
  98. Topological invariants for the line graphs of some classes of graphs
  99. Words for maximal Subgroups of Fi24
  100. Generators of Maximal Subgroups of Harada-Norton and some Linear Groups
  101. Special Issue on POKOCHA 2018
  102. Influence of Production Parameters on the Content of Polyphenolic Compounds in Extruded Porridge Enriched with Chokeberry Fruit (Aronia melanocarpa (Michx.) Elliott)
  103. Effects of Supercritical Carbon Dioxide Extraction (SC-CO2) on the content of tiliroside in the extracts from Tilia L. flowers
  104. Impact of xanthan gum addition on phenolic acids composition and selected properties of new gluten-free maize-field bean pasta
  105. Impact of storage temperature and time on Moldavian dragonhead oil – spectroscopic and chemometric analysis
  106. The effect of selected substances on the stability of standard solutions in voltammetric analysis of ascorbic acid in fruit juices
  107. Determination of the content of Pb, Cd, Cu, Zn in dairy products from various regions of Poland
  108. Special Issue on IC3PE 2018 Conference
  109. The Photocatalytic Activity of Zns-TiO2 on a Carbon Fiber Prepared by Chemical Bath Deposition
  110. N-octyl chitosan derivatives as amphiphilic carrier agents for herbicide formulations
  111. Kinetics and Mechanistic Study of Hydrolysis of Adenosine Monophosphate Disodium Salt (AMPNa2) in Acidic and Alkaline Media
  112. Antimalarial Activity of Andrographis Paniculata Ness‘s N-hexane Extract and Its Major Compounds
  113. Special Issue on ABB2018 Conference
  114. Special Issue on ICCESEN 2017
  115. Theoretical Diagnostics of Second and Third-order Hyperpolarizabilities of Several Acid Derivatives
  116. Determination of Gamma Rays Efficiency Against Rhizoctonia solani in Potatoes
  117. Studies On Compatibilization Of Recycled Polyethylene/Thermoplastic Starch Blends By Using Different Compatibilizer
  118. Liquid−Liquid Extraction of Linalool from Methyl Eugenol with 1-Ethyl-3-methylimidazolium Hydrogen Sulfate [EMIM][HSO4] Ionic Liquid
  119. Synthesis of Graphene Oxide Through Ultrasonic Assisted Electrochemical Exfoliation
  120. Special Issue on ISCMP 2018
  121. Synthesis and antiproliferative evaluation of some 1,4-naphthoquinone derivatives against human cervical cancer cells
  122. The influence of the grafted aryl groups on the solvation properties of the graphyne and graphdiyne - a MD study
  123. Electrochemical modification of platinum and glassy carbon surfaces with pyridine layers and their use as complexing agents for copper (II) ions
  124. Effect of Electrospinning Process on Total Antioxidant Activity of Electrospun Nanofibers Containing Grape Seed Extract
  125. Effect Of Thermal Treatment Of Trepel At Temperature Range 800-1200˚C
  126. Topical Issue on Agriculture
  127. The effect of Cladophora glomerata exudates on the amino acid composition of Cladophora fracta and Rhizoclonium sp.
  128. Influence of the Static Magnetic Field and Algal Extract on the Germination of Soybean Seeds
  129. The use of UV-induced fluorescence for the assessment of homogeneity of granular mixtures
  130. The use of microorganisms as bio-fertilizers in the cultivation of white lupine
  131. Lyophilized apples on flax oil and ethyl esters of flax oil - stability and antioxidant evaluation
  132. Production of phosphorus biofertilizer based on the renewable materials in large laboratory scale
  133. Human health risk assessment of potential toxic elements in paddy soil and rice (Oryza sativa) from Ugbawka fields, Enugu, Nigeria
  134. Recovery of phosphates(V) from wastewaters of different chemical composition
  135. Special Issue on the 4th Green Chemistry 2018
  136. Dead zone for hydrogenation of propylene reaction carried out on commercial catalyst pellets
  137. Improved thermally stable oligoetherols from 6-aminouracil, ethylene carbonate and boric acid
  138. The role of a chemical loop in removal of hazardous contaminants from coke oven wastewater during its treatment
  139. Combating paraben pollution in surface waters with a variety of photocatalyzed systems: Looking for the most efficient technology
  140. Special Issue on Chemistry Today for Tomorrow 2019
  141. Applying Discriminant and Cluster Analyses to Separate Allergenic from Non-allergenic Proteins
  142. Chemometric Expertise Of Clinical Monitoring Data Of Prolactinoma Patients
  143. Chemomertic Risk Assessment of Soil Pollution
  144. New composite sorbent for speciation analysis of soluble chromium in textiles
  145. Photocatalytic activity of NiFe2O4 and Zn0.5Ni0.5Fe2O4 modified by Eu(III) and Tb(III) for decomposition of Malachite Green
  146. Photophysical and antibacterial activity of light-activated quaternary eosin Y
  147. Spectral properties and biological activity of La(III) and Nd(III) Monensinates
  148. Special Issue on Monitoring, Risk Assessment and Sustainable Management for the Exposure to Environmental Toxins
  149. Soil organic carbon mineralization in relation to microbial dynamics in subtropical red soils dominated by differently sized aggregates
  150. A potential reusable fluorescent aptasensor based on magnetic nanoparticles for ochratoxin A analysis
  151. Special Issue on 13th JCC 2018
  152. Fluorescence study of 5-nitroisatin Schiff base immobilized on SBA-15 for sensing Fe3+
  153. Thermal and Morphology Properties of Cellulose Nanofiber from TEMPO-oxidized Lower part of Empty Fruit Bunches (LEFB)
  154. Encapsulation of Vitamin C in Sesame Liposomes: Computational and Experimental Studies
  155. A comparative study of the utilization of synthetic foaming agent and aluminum powder as pore-forming agents in lightweight geopolymer synthesis
  156. Synthesis of high surface area mesoporous silica SBA-15 by adjusting hydrothermal treatment time and the amount of polyvinyl alcohol
  157. Review of large-pore mesostructured cellular foam (MCF) silica and its applications
  158. Ion Exchange of Benzoate in Ni-Al-Benzoate Layered Double Hydroxide by Amoxicillin
  159. Synthesis And Characterization Of CoMo/Mordenite Catalyst For Hydrotreatment Of Lignin Compound Models
  160. Production of Biodiesel from Nyamplung (Calophyllum inophyllum L.) using Microwave with CaO Catalyst from Eggshell Waste: Optimization of Transesterification Process Parameters
  161. The Study of the Optical Properties of C60 Fullerene in Different Organic Solvents
  162. Composite Material Consisting of HKUST-1 and Indonesian Activated Natural Zeolite and its Application in CO2 Capture
  163. Topical Issue on Environmental Chemistry
  164. Ionic liquids modified cobalt/ZSM-5 as a highly efficient catalyst for enhancing the selectivity towards KA oil in the aerobic oxidation of cyclohexane
  165. Application of Thermal Resistant Gemini Surfactants in Highly Thixotropic Water-in-oil Drilling Fluid System
  166. Screening Study on Rheological Behavior and Phase Transition Point of Polymer-containing Fluids produced under the Oil Freezing Point Temperature
  167. The Chemical Softening Effect and Mechanism of Low Rank Coal Soaked in Alkaline Solution
  168. The Influence Of NO/O2 On The NOx Storage Properties Over A Pt-Ba-Ce/γ-Al2O3 Catalyst
  169. Special Issue on the International conference CosCI 2018
  170. Design of SiO2/TiO2 that Synergistically Increases The Hydrophobicity of Methyltrimethoxysilane Coated Glass
  171. Antidiabetes and Antioxidant agents from Clausena excavata root as medicinal plant of Myanmar
  172. Development of a Gold Immunochromatographic Assay Method Using Candida Biofilm Antigen as a Bioreceptor for Candidiasis in Rats
  173. Special Issue on Applied Biochemistry and Biotechnology 2019
  174. Adsorption of copper ions on Magnolia officinalis residues after solid-phase fermentation with Phanerochaete chrysosporium
  175. Erratum
  176. Erratum to: Sand Dune Characterization For Preparing Metallurgical Grade Silicon
Heruntergeladen am 5.10.2025 von https://www.degruyterbrill.com/document/doi/10.1515/chem-2019-0015/html
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