ABSTRACT
Orlistat has been proved to be an effective fatty acid synthase inhibitor that is able to inhibit the proliferation and induce apoptosis in many cancer cell types. However, the anticancer effects of orlistat on hepatocellular carcinoma are undefined. We found that orlistat inhibited cell growth and induced G0/G1 cell cycle arrest with increased cyclin D, cyclin E, and p21 expression in human hepatoma Hep3B cells. Furthermore, protein expression of cyclin A, cyclin B, Cdk1, Cdk2, and Cdk4 was reduced by orlistat. This study investigated the role of lipid metabolism on orlistat-induced human hepatoma Hep3B cell death. The decrease in the expression of key enzymes in fatty acid metabolism, including FASN, ACOT8, PPT1, FABP1, CPT1 and CPT2, was observed after orlistat treatment. We also demonstrated that peroxisomal activity was involved in the orlistat-induced Hep3B cell death. In this study, we established an in vitro model to investigate the effect of orlistat on lipid accumulation. We found that orlistat significantly inhibited the cellular lipid content when administered in fatty acid overload conditions in Hep3B cells. Combination treatment of orlistat and paclitaxel was able to induce a synergistic effect on growth inhibition and cell apoptosis in Hep3B cells. Our data suggested that orlistat displays antitumor activity and enhances the efficacy of paclitaxel in Hep3B cells.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Orlistat/pharmacology , Paclitaxel/pharmacology , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Cell Proliferation/drug effects , Drug Synergism , G1 Phase Cell Cycle Checkpoints/drug effects , Humans , Lipid Metabolism/genetics , Liver Neoplasms/pathology , Palmitoyl-CoA Hydrolase/genetics , Palmitoyl-CoA Hydrolase/metabolism , Peroxisomes/metabolismABSTRACT
We set out studies on anion- and solvent-induced assembly based on the ligand N-(4-(4-aminophenyloxy)phenyl)isonicotinamide (papoa), which is synthesized to show a bent and flexible backbone. Reactions of papoa with ZnX2 (X=Cl, Br, and I) gave the dinuclear macrocycles ([ZnX2 (papoa)]2 ; X=Cl (1 a), Br (2 a), I (3)), the structure of which was determined by X-ray diffraction. Notably, the less bulky Cl and Br compounds afforded the coordinated imine in acetone (i.e., [ZnX2 (papoi)]2 , papoi=N-(4-(4-(propan-2-ylideneamino)phenoxy)phenyl)isonicotinamide; X=Cl (1 b), Br (2 b)), whereas the iodine one only gave the coordinated amine compound 3 under the same reaction condition. In fact, the coordinated imine can return to the amine analogue upon exposure to air or in DMSO, which has been monitored by (1)Hâ NMR spectroscopy and powder X-ray diffraction. Both the dinuclear [Zn(papoa)(NO3)2]2 (4 a) and the 1D [Zn(papoa)2(NO3)2]n (4 b) were formed from the reaction of Zn(NO3)2 and papoa in mixed solvents with acetone and acetonitrile, respectively. In addition, Cd(ClO4)2 can react with papoa to give the 1D framework {[Cd(papoa)2 (CH3CN)2](ClO4)2}n (5 a) and the 2D framework [Cd(papoa)2 (ClO4)2]n (5 b), depending on the solvent used, that is, MeOH and CH3CN, respectively. Importantly, the 1D framework with axially coordinated CH3 CN molecules and the 2D framework with axially coordinated ClO4(-) ions can be interconverted by heating and grinding in the presence of CH3CN, respectively. Such a reversible structural transformation process was proven by PXRD studies.
ABSTRACT
This study investigated methanogenic communities involved in degradation of tetramethylammonium hydroxide (TMAH) in three full-scale bioreactors treating TMAH-containing wastewater. Based on the results of terminal-restriction fragment-length polymorphism (T-RFLP) and quantitative PCR analyses targeting the methyl-coenzyme M reductase alpha subunit (mcrA) genes retrieved from three bioreactors, Methanomethylovorans and Methanosarcina were the dominant methanogens involved in the methanogenic degradation of TMAH in the bioreactors. Furthermore, batch experiments were conducted to evaluate mcrA messenger RNA (mRNA) expression during methanogenic TMAH degradation, and the results indicated that a higher level of TMAH favored mcrA mRNA expression by Methansarcina, while Methanomethylovorans could only express considerable amount of mcrA mRNA at a lower level of TMAH. These results suggest that Methansarcina is responsible for methanogenic TMAH degradation at higher TMAH concentrations, while Methanomethylovorans may be important at a lower TMAH condition.
Subject(s)
Bacteria/metabolism , Bioreactors/microbiology , Methane/metabolism , Quaternary Ammonium Compounds/metabolism , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Polymorphism, Restriction Fragment Length , Sewage/microbiologyABSTRACT
A series of well-defined tetracosanuclear nickel complexes 3-7 facilely produced by one-pot synthesis were active catalysts for cycloaddition of CO2 and cyclohexene oxide (CHO). These nickel complexes were doughnut-like supramolecular coordination complexes involving eight repeating units, and each of them contains one Schiff base ligand and three nickel(II) ions. Notably, the 24-nuclear nickel cluster complex 3 in combination with nucleophilic additives was the most efficient catalyst to mediate CO2 coupling with CHO to generate CO2-based cis-cyclohexene carbonates. In addition to CO2/CHO cycloaddition, complex 3 was also found to effectively couple CO2 with other alicyclic epoxides, generating the corresponding cyclic carbonates. Additionally, kinetic investigations for CO2 cycloaddition of CHO using 3 were reported.
ABSTRACT
This study aimed to determine the free radical scavenging and antioxidant potential of hot water extracts prepared from different combinations and ratios of submerged cultivated mycelial biomass of medicinal mushrooms. Total phenolic compounds, flavonoid content, and antioxidant activity were evaluated for combined crude hot water extracts from medicinal higher Basidiomycetes mushrooms belonging to ten genera. The results demonstrate that almost all tested combinations were good sources of phenolic compounds and flavonoids, ranging between 16.42 and 18.83 gallic acid equivalents/g and 1.5 and 4.34 mg rutin equivalents/g, respectively. Moreover, free radical scavenging properties were evaluated with the DPPH and ABTS assays and metal chelating effects were investigated. All tested samples and/or extracts demonstrated significant free radical scavenging properties and antioxidant potential.
Subject(s)
Agaricales , Antioxidants , Agaricales/chemistry , Antioxidants/chemistry , Biomass , Flavonoids/chemistry , Free Radicals , Phenols/analysis , Plant Extracts/chemistry , WaterABSTRACT
This research describes the investigation of submerged cultivated mycelial biomass and hot water extracts prepared from different combinations and ratios of medicinal mushroom (MM) dry powders, comprising various biologically active compounds/secondary metabolites. In particular, it was evaluated the proximate composition (moisture, ash, crude protein, fat, total carbohydrates, and total energy), γ-aminobutyric acid (GABA) and ergothioneine (ERG), amino acid content of mycelia of 16 higher Basidiomycetes MM species. The results obtained demonstrate that almost all tested combinations were found to be good sources of polysaccharides, with content varying in the ranges of 4.73 ± 1.33% and 58.46 ± 4.15%. Total protein contents varied in 1.97 ± 0.40% - 5.37 ± 0.40% range. ERG was detected in all tested samples, while GABA existed only in eight samples out of 15 and varied from 0.03 ± < 0.01 to 0.61 ± 0.03 mg/g, and from 0.16 ± 0.03 to 5.69 ± 0.41 mg/g respectively. Analyses of total phenolic and flavonoid contents demonstrate considerable content in all samples (15.53 ± 0.23 - 18.88 ± 0.34 mg gallic acid equivalents/g and 1.23 ± 0.04 - 4.34 ± 0.73 mg rutin equivalents/g respectively). In present research the complexity of samples/extracts were evaluated by multiple antioxidant assays to verify their antioxidant capacity. Determination of in vitro antioxidant activity was successfully carried out by several different methods such as 2,2-diphenyl-1-picrylhydrazyl scavenging activity, reducing power, chelating ability, hydroxyl radical scavenging activity, and 2,2'-azino-bis(3-ethylbenzothi-azoline-6-sulfonic acid scavenging activity. Therefore, all tested samples confirm the capable antioxidant activities of bioactive compounds extracted from MMs.
Subject(s)
Agaricales , Antioxidants , Flavonoids , Mycelium , PhenolsABSTRACT
Nontarget data acquisition for target analysis (nDATA) workflows using liquid chromatography-high-resolution accurate mass (LC-HRAM) spectrometry, spectral screening software, and a compound database have generated interest because of their potential for screening of pesticides in foods. However, these procedures and particularly the instrument processing software need to be thoroughly evaluated before implementation in routine analysis. In this work, 25 laboratories participated in a collaborative study to evaluate an nDATA workflow on high moisture produce (apple, banana, broccoli, carrot, grape, lettuce, orange, potato, strawberry, and tomato). Samples were extracted in each laboratory by quick, easy, cheap, effective, rugged, and safe (QuEChERS), and data were acquired by ultrahigh-performance liquid chromatography (UHPLC) coupled to a high-resolution quadrupole Orbitrap (QOrbitrap) or quadrupole time-of-flight (QTOF) mass spectrometer operating in full-scan mass spectrometry (MS) data-independent tandem mass spectrometry (LC-FS MS/DIA MS/MS) acquisition mode. The nDATA workflow was evaluated using a restricted compound database with 51 pesticides and vendor processing software. Pesticide identifications were determined by retention time (tR, ±0.5 min relative to the reference retention times used in the compound database) and mass errors (δM) of the precursor (RTP, δM ≤ ±5 ppm) and product ions (RTPI, δM ≤ ±10 ppm). The elution profiles of all 51 pesticides were within ±0.5 min among 24 of the participating laboratories. Successful screening was determined by false positive and false negative rates of <5% in unfortified (pesticide-free) and fortified (10 and 100 µg/kg) produce matrices. Pesticide responses were dependent on the pesticide, matrix, and instrument. The false negative rates were 0.7 and 0.1% at 10 and 100 µg/kg, respectively, and the false positive rate was 1.1% from results of the participating LC-HRAM platforms. Further evaluation was achieved by providing produce samples spiked with pesticides at concentrations blinded to the laboratories. Twenty-two of the 25 laboratories were successful in identifying all fortified pesticides (0-7 pesticides ranging from 5 to 50 µg/kg) for each produce sample (99.7% detection rate). These studies provide convincing evidence that the nDATA comprehensive approach broadens the screening capabilities of pesticide analyses and provide a platform with the potential to be easily extended to a larger number of other chemical residues and contaminants in foods.
Subject(s)
Pesticide Residues , Pesticides , Chromatography, High Pressure Liquid , Chromatography, Liquid , Food Contamination/analysis , Fruit/chemistry , Pesticide Residues/analysis , Pesticides/analysis , Tandem Mass Spectrometry , Vegetables , WorkflowABSTRACT
Recent studies indicate that neonatal spermatogonial stem cells (SSCs) possess pluripotency. However, the mechanisms that regulate the pluripotent differentiation capacity of SSCs remain unclear. Here, we describe a new method to clonally derive pluripotent SSCs from neonatal mouse testis. By coculturing with testicular stromal cells, SSCs can be maintained and expanded in serum-free conditions. Unlike endogenous SSCs, these in vitro expanded SSCs showed strong alkaline phosphatase (AP) activity and displayed characteristics of embryonic stem cells and primordial germ cells, which were therefore designated as AP(+) germline stem cells (AP(+)GSCs). The pluripotency of AP(+)GSCs was confirmed by in vitro differentiation toward hepatic and neuronal lineages and formation of embryonic chimeras after injection into blastocysts. Further investigation revealed that insulin-like growth factor-1 (IGF-1) secreted from Leydig cells was a key factor involved in maintaining the pluripotency of AP(+)GSCs. The blockage of IGF-1 receptor phosphorylation and its downstream PI3K pathway by PPP or LY294002 dramatically reduced their AP activity and expression of pluripotent genes, such as Oct-4, Blimp1, and Nanog. In conclusion, the present study demonstrated that IGF-1 secreted by testicular Leydig cells plays an important role in maintaining the pluripotency of SSCs in culture, which provides an insight into the molecular mechanism underlying germ cell pluripotency.
Subject(s)
Insulin-Like Growth Factor I/metabolism , Pluripotent Stem Cells/cytology , Spermatogonia/cytology , Animals , Animals, Newborn , Cell Lineage , Insulin-Like Growth Factor I/physiology , Leydig Cells/metabolism , Male , Mice , Paracrine Communication , Testis/cytologyABSTRACT
Fibroblast growth factor receptor 4 (FGFR4) is involved in multiple physiological and pathological processes. Several genetic variants of FGFR4 have been shown to be associated with tumor progression in many cancers. However, its association, such as genetic variants and expression levels, with lung cancer is controversial. The present study examined the relationship between four single-nucleotide polymorphisms (SNPs; rs2011077 T/C, rs351855 G/A, rs7708357 G/A, and rs1966265 A/G) of FGFR4 and the risk of lung adenocarcinoma with the epidermal growth factor receptor (EGFR) mutation status in a Taiwanese cohort. The results demonstrated that FGFR4 rs2011077 (odds ratio (OR) = 0.348, 95% confidence interval (CI) = 0.136-0.891, p = 0.024), and rs351855 (OR = 0.296, 95% CI = 0.116-0.751, p = 0.008) showed an inverse association with distant metastasis in wild-type EGFR lung adenocarcinoma. Furthermore, a database analysis using The Cancer Genome Atlas revealed that the higher FGFR4 expression level was correlated with poor survival rates in wild-type EGFR lung adenocarcinoma. In conclusion, the data suggest that FGFR4 SNPs may help in identifying patient subgroups at low-risk for tumor metastasis, among carriers of lung adenocarcinoma bearing wild-type EGFR.
Subject(s)
Adenocarcinoma of Lung/genetics , Lung Neoplasms/genetics , Receptor, Fibroblast Growth Factor, Type 4/genetics , Adenocarcinoma of Lung/pathology , Female , Genetic Predisposition to Disease , Humans , Lung Neoplasms/pathology , Male , Polymorphism, Single Nucleotide , Taiwan/epidemiologyABSTRACT
Hepatic fibrosis may ultimately result in organ failure and death, a reality compounded by the fact that most drugs for liver fibrosis appear to be effective only if given as a prophylactic or early treatment. In a dimethylnitrosamine-induced liver fibrotic model, aspartate aminotransferase/alanine aminotransferase levels could not precisely distinguish the differences between the initial stage of liver fibrosis and normal control, whereas histological examination indicated that dimethylnitrosamine treatment for two weeks has resulted in hepatic fibrogenesis. Comprehensive proteomics identified 12 proteins mainly associated with the interleukin 6-stimulated inflammatory pathway. Coordinately, cytokine profiles showed that dimethylnitrosamine administration would stimulate various signaling pathways leading to liver fibrosis. Of note, apolipoprotein A4 in serum samples obtained from patients in the early stage of liver fibrosis were significantly increased compared to the healthy controls (p<0.001) while the area under curve was 0.966. Moreover, increased apolipoprotein A4 significantly enhanced transforming growth factor beta 1-induced alpha smooth muscle actin expression. In this regard, overexpression of apolipoprotein A4 in early stage of liver fibrosis might magnify and imply the progression of hepatic fibrosis. These findings suggest that apolipoprotein A4 upregulation may correlate with hepatic fibrosis staging and that apolipoprotein A4 together with current biomarker can increase the sensitivity and specificity for the early detection of liver fibrosis in a high-throughput manner.
ABSTRACT
This study evaluated biological treatment of TMAH in a full-scale methanogenic up-flow anaerobic sludge blanket (UASB) followed by an aerobic bioreactor. In general, the UASB was able to perform a satisfactory TMAH degradation efficiency, but the effluent COD of the aerobic bioreactor seemed to increase with an increased TMAH in the influent wastewater. The batch test results confirmed that the UASB sludge under methanogenic conditions would be favored over the aerobic ones for TMAH treatment due to its superb ability of handling high strength of TMAH-containing wastewaters. Based on batch experiments, inhibitory chemicals present in TFT-LCD wastewater like surfactants and sulfate should be avoided to secure a stable methanogenic TMAH degradation. Finally, molecular monitoring of Methanomethylovorans hollandica and Methanosarcina mazei in the full-scale plant, the dominant methanogens in the UASB responsible for TMAH degradation, may be beneficial for a stable TMAH treatment performance.
Subject(s)
Hydroxides/metabolism , Water Pollutants, Chemical/isolation & purification , Ammonium Hydroxide , Base Sequence , DNA Primers , Polymerase Chain Reaction , Polymorphism, Restriction Fragment LengthABSTRACT
Oocytes fertilized with spermatozoa obtained from Septin 12+/- chimeric mice failed to develop beyond the morula stage after IVF and intracytoplasmic sperm injection because of significant DNA defects in the spermatozoa. Given that SEPT12 is expressed at the edge of the sperm nucleus in both humans and mice, we hypothesized the vital roles of Septin 12 in sperm head shaping, nuclear DNA condensation, and early embryonic development.