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1.
Eur J Clin Invest ; 53(4): e13929, 2023 Apr.
Article in English | MEDLINE | ID: mdl-36453878

ABSTRACT

BACKGROUND: Oesophageal carcinoma (EC) is one of the types of prevalent malignant cancer in the globe. Many researchers reported the vital role played by long-coding RNAs in EC. In the current research, we investigated the mechanisms of the action of lncRNA BBOX1-AS1 in EC progression. METHODS: In EC tissues and EC cells, the expression levels of miR-361-3p along with COL1A1 and BBOX1-AS1 were detected through RT-qPCR or western blotting. MiR-361-3p interactions with BBOX1-AS1 or COL1A1 were verified through Luciferase reporter and RIP tests. Loss of function combined with caspase-3 activity, CCK-8 and Transwell assays was performed to investigate cell apoptosis, proliferation and migration, respectively. Knockdown of BBOX1-AS1 was used for evaluating BBOX1-AS1 effects on tumour development in vivo. RESULTS: BBOX1-AS1 was remarkably elevated in EC tissues and cells. In addition, the silencing of BBOX1-AS1 attenuated the cell viability, cell migration and enhanced cell apoptosis of EC, as well as suppressed EC tumour formation in vivo. Moreover, BBOX1-AS1 was found to be a sponge of miR-361-3p, which downregulated miR-361-3p expression. MiR-361-3p inhibitor rescued the anti-tumour effect of BBOX1-AS1 knockdown on the progression of EC. Furthermore, we discovered that miR-361-3p specially bound to COL1A1 3'UTR and downregulated COL1A1 and COL1A1 reduction declined the promoting effect of silencing miR-361-3p on EC cell malignant phenotypes. CONCLUSION: BBOX1-AS1 facilitated the EC development and malignancy via miR-361-3p/COL1A1 axis, indicating BBOX1-AS1 could be a novel therapy target for the diagnostic of EC.


Subject(s)
Esophageal Neoplasms , MicroRNAs , RNA, Long Noncoding , Humans , Apoptosis , Blotting, Western , Cell Movement , Cell Proliferation , Cell Line, Tumor , Gene Expression Regulation, Neoplastic
2.
Electrophoresis ; 43(11): 1215-1222, 2022 06.
Article in English | MEDLINE | ID: mdl-35286725

ABSTRACT

Protein therapeutics are usually produced in heterogeneous forms during bioproduction and bioprocessing. Heterogeneity results from post-translational modifications that can yield charge variants and require characterization throughout product development and manufacturing. Isoelectric focusing (IEF) with UV detection is one of the most common methods to evaluate protein charge heterogeneity in the biopharmaceutical industry. To identify charge variant peaks, a new imaged microfluidic chip-based isoelectric focusing (icIEF) system coupled directly to mass spectrometry was recently reported. Bridging is required to demonstrate comparability between existing and new technology. As such, here we demonstrate the comparability of the pI value measurement and relative charge species distributions between the icIEF-MS system and the control data from a frequently utilized methodology in the biopharmaceutical industry for several blinded development-phase biopharmaceutical monoclonal antibodies across a wide pI range of 7.3-9.0. Hyphenation of the icIEF system with mass spectrometry enabled direct and detailed structural determination of a test molecule, with masses suggesting acidic and basic shifts are caused by sialic acid additions and the presence of unprocessed lysine residues. In addition, MS analysis further identified several low-abundance glycoforms. The icIEF-MS system provides sample quantification, characterization, and identification of mAb proteoforms without sacrificing icIEF quantification comparability or speed.


Subject(s)
Biological Products , Electrophoresis, Capillary , Antibodies, Monoclonal/analysis , Electrophoresis, Capillary/methods , Isoelectric Focusing/methods , Mass Spectrometry/methods
3.
Electrophoresis ; 43(9-10): 1050-1058, 2022 05.
Article in English | MEDLINE | ID: mdl-35245390

ABSTRACT

An international team spanning 19 sites across 18 biopharmaceutical and in vitro diagnostics companies in the United States, Europe, and China, along with one regulatory agency, was formed to compare the precision and robustness of imaged CIEF (ICIEF) for the charge heterogeneity analysis of the National Institute of Standards and Technology (NIST) mAb and a rhPD-L1-Fc fusion protein on the iCE3 and the Maurice instruments. This information has been requested to help companies better understand how these instruments compare and how to transition ICIEF methods from iCE3 to the Maurice instrument. The different laboratories performed ICIEF on the NIST mAb and rhPD-L1-Fc with both the iCE3 and Maurice using analytical methods specifically developed for each of the molecules. After processing the electropherograms, statistical evaluation of the data was performed to determine consistencies within and between laboratory and outlying information. The apparent isoelectric point (pI) data generated, based on two-point calibration, for the main isoform of the NIST mAb showed high precision between laboratories, with RSD values of less than 0.3% on both instruments. The SDs for the NIST mAb and the rhPD-L1-Fc charged variants percent peak area values for both instruments are less than 1.02% across different laboratories. These results validate the appropriate use of both the iCE3 and Maurice for ICIEF in the biopharmaceutical industry in support of process development and regulatory submissions of biotherapeutic molecules. Further, the data comparability between the iCE3 and Maurice illustrates that the Maurice platform is a next-generation replacement for the iCE3 that provides comparable data.


Subject(s)
Biological Products , Electrophoresis, Capillary , Electrophoresis, Capillary/methods , Isoelectric Focusing/methods , Laboratories , Protein Isoforms
4.
J Environ Manage ; 321: 115834, 2022 Nov 01.
Article in English | MEDLINE | ID: mdl-35994959

ABSTRACT

This study proposes an equitable and effective water resource planning framework that handles competing regions and conflicting water departments within water-stressed watersheds under uncertainty. To cope with uncertainty, a robust optimization method based on an ellipsoidal uncertainty set is presented to keep the best solution viable and less conservative while attempting to find a balance between the reliability and optimum goals. The comprehensive framework consists of two predominant steps: equitable initial water allocation robust optimization programming, which employs the absolute difference between the supply and demand of water resources as the objective function and the Gini coefficient as the restricted condition, given that both surface water and groundwater are random, and subsequent effective water re-allocation robust modeling, which applies cooperative game theory to achieve maximum welfare in a river basin under uncertainty of the benefit coefficient. A realistic example of the Tuojiang River Basin was conducted, and the simulation experiments showed a significant increase (79.20%) in reliability with only an 11.44% increase in objective values and 2.76% and 15.17% decreases in the amount of surface water and groundwater, respectively. The findings of the analysis reveal that robust policies achieve adaptive optimal decision-making under uncertainty at a low cost.


Subject(s)
Rivers , Water Resources , Reproducibility of Results , Water/analysis , Water Pollution/analysis , Water Supply
5.
EMBO J ; 36(10): 1330-1347, 2017 05 15.
Article in English | MEDLINE | ID: mdl-28377463

ABSTRACT

Pluripotent stem cells are known to display distinct metabolic phenotypes than their somatic counterparts. While accumulating studies are focused on the roles of glucose and amino acid metabolism in facilitating pluripotency, little is known regarding the role of lipid metabolism in regulation of stem cell activities. Here, we show that fatty acid (FA) synthesis activation is critical for stem cell pluripotency. Our initial observations demonstrated enhanced lipogenesis in pluripotent cells and during cellular reprogramming. Further analysis indicated that de novo FA synthesis controls cellular reprogramming and embryonic stem cell pluripotency through mitochondrial fission. Mechanistically, we found that de novo FA synthesis regulated by the lipogenic enzyme ACC1 leads to the enhanced mitochondrial fission via (i) consumption of AcCoA which affects acetylation-mediated FIS1 ubiquitin-proteasome degradation and (ii) generation of lipid products that drive the mitochondrial dynamic equilibrium toward fission. Moreover, we demonstrated that the effect of Acc1 on cellular reprogramming via mitochondrial fission also exists in human iPSC induction. In summary, our study reveals a critical involvement of the FA synthesis pathway in promoting ESC pluripotency and iPSC formation via regulating mitochondrial fission.


Subject(s)
Fatty Acids/metabolism , Mitochondrial Dynamics , Pluripotent Stem Cells/metabolism , Acetyl Coenzyme A/metabolism , Acetyl-CoA Carboxylase/metabolism , Humans
6.
Mol Ecol ; 30(13): 3158-3174, 2021 07.
Article in English | MEDLINE | ID: mdl-33481325

ABSTRACT

Environmental DNA (eDNA) metabarcoding can rapidly characterize the composition and diversity of benthic communities, thus it has high potential utility for routine assessments of benthic impacts of marine finfish farming. In this study, 126 sediment grab samples from 42 stations were collected at six salmon farms in British Columbia, Canada. Benthic community changes were assessed by both eDNA metabarcoding of metazoans and macrofaunal polychaete surveys. The latter was done by analysing 11,466 individuals using a combination of morphology-based taxonomy and DNA barcoding. Study objectives were to: (i) compare biotic signals associated with benthic impacts of salmon farming in the two data sources, and (ii) identify potential eDNA indicators to facilitate monitoring in Canada. Alpha diversity parameters were consistently reduced near fish cage edge and negatively correlated with pore-water sulphide concentration, with coefficients ranging from -0.62 to -0.48. Although Polychaeta are a common indicator group, the negative correlation with pore-water sulphide concentration was much stronger for Nematoda OTU richness (correlation coefficient: -0.86) than for Polychaeta (correlation coefficient: -0.38). Presence/absence of Capitella generally agreed well between the two methods despite that they differed in the volume of sediments sampled and the molecular marker used. Multiple approaches were used to identify OTUs related to organic enrichment statuses. We demonstrate that eDNA metabarcoding generates biotic signals that could be leveraged for environmental assessment of benthic impacts of fish farms in multiple ways: both alpha diversity and Nematoda OTU richness could be used to assess the spatial extent of impact, and OTUs related to organic enrichment could be used to develop local biotic indices.


Subject(s)
DNA Barcoding, Taxonomic , Salmon , Animals , Aquaculture , Biodiversity , British Columbia , Environmental Monitoring , Geologic Sediments , Humans , Salmon/genetics
7.
EMBO Rep ; 20(10): e48115, 2019 10 04.
Article in English | MEDLINE | ID: mdl-31379107

ABSTRACT

Lin28 plays an important role in promoting tumor development, whereas its exact functions and underlying mechanisms are largely unknown. Here, we show that both human homologs of Lin28 accelerate de novo fatty acid synthesis and promote the conversion from saturated to unsaturated fatty acids via the regulation of SREBP-1. By directly binding to the mRNAs of both SREBP-1 and SCAP, Lin28A/B enhance the translation and maturation of SREBP-1, and protect cancer cells from lipotoxicity. Lin28A/B-stimulated tumor growth is abrogated by SREBP-1 inhibition and by the impairment of the RNA binding properties of Lin28A/B, respectively. Collectively, our findings uncover that post-transcriptional regulation by Lin28A/B enhances de novo fatty acid synthesis and metabolic conversion of saturated and unsaturated fatty acids via SREBP-1, which is critical for cancer progression.


Subject(s)
Disease Progression , Fatty Acids/biosynthesis , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , RNA-Binding Proteins/metabolism , Sterol Regulatory Element Binding Protein 1/metabolism , Cell Line, Tumor , Cell Proliferation , Cytoprotection , Endoplasmic Reticulum Stress , Humans , Intracellular Signaling Peptides and Proteins/genetics , Intracellular Signaling Peptides and Proteins/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Models, Biological , Protein Binding , Protein Biosynthesis , RNA, Messenger/genetics , RNA, Messenger/metabolism
8.
Int J Syst Evol Microbiol ; 71(12)2021 Dec.
Article in English | MEDLINE | ID: mdl-34913427

ABSTRACT

An aerobic Gram-stain-negative, curved rod-shaped and non-spore-forming bacterial strain (NBU2194T) was isolated from seawater collected in an intertidal zone in Ningbo, Zhejiang Province, PR China. It was motile though a single polar flagellum and grew at 20-42 °C (optimum, 30 °C), in 0-2.0 % NaCl (0 %, w/v) and at pH 5.0-9.0 (pH 6.0-7.0). The sole respiratory quinone was ubiquinone-8. The major cellular fatty acids were C16 : 0, C16 : 1 ω7c and/or C16 : 1 ω6c. The polar lipids contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, one unidentified phospholipid and two unidentified aminophosphoglycolipids. A phylogenetic analysis based on 16S rRNA gene sequences and 65 genomic core genes showed that strain NBU2194T formed a distinct lineage in the family Alteromonadaceae. The genome of strain NBU2194T was 4 913 533 bp with a DNA G+C content of 43.9 mol% and coded 3895 genes, 12 rRNA genes and 47 tRNA genes. The average nucleotide identity, amino acid identity and digital DNA-DNA hybridization values between strain NBU2194T and related species of Alteromonadaceae were below the threshold limit for prokaryotic species delineation. NBU2194T could be distinguished from other genera in the family Alteromonadaceae based on phenotypic, chemotaxonomic and genomic characteristics. On the basis of the polyphasic taxonomic evidence collected in this study, strain NBU2194T is considered to represent a novel genus and species in the family Alteromonadaceae, for which the name Paraneptunicella aestuarii is proposed. The type strain is NBU2194T (=KCTC 82442T=GDMCC 1.2217T).


Subject(s)
Alteromonadaceae , Phylogeny , Seawater/microbiology , Alteromonadaceae/classification , Alteromonadaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/chemistry , Nucleic Acid Hybridization , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
9.
Zhongguo Zhong Yao Za Zhi ; 46(23): 6224-6230, 2021 Dec.
Article in Zh | MEDLINE | ID: mdl-34951249

ABSTRACT

Alzheimer's disease(AD) patients in China have been surging, and the resultant medical burden and care demand have a huge impact on the development of individuals, families, and the society. The active component compound of Epimedii Folium, Astragali Radix, and Puerariae Lobatae Radix(YHG) can regulate the expression of iron metabolism-related proteins to inhibit brain iron overload and relieve hypofunction of central nervous system in AD patients. Hepcidin is an important target regulating iron metabolism. This study investigated the effect of YHG on the expression of a disintegrin and metalloprotease-17(ADAM17), a key enzyme in the hydrolysis of ß amyloid precursor protein(APP) in HT22 cells, by mediating hepcidin. To be specific, HT22 cells were cultured in vitro, followed by liposome-mediated siRNA transfection to silence the expression of hepcidin. Real-time PCR and Western blot were performed to examine the silencing result and the effect of YHG on hepcidin in AD cell model. HT22 cells were randomized into 7 groups: control group, Aß25-35 induction(Aß) group, hepcidin-siRNA(siRNA) group, Aß25-35 + hepcidin-siRNA(Aß + siRNA) group, Aß25-35+YHG(Aß+YHG) group, hepcidin-siRNA+YHG(siRNA+YHG) group, Aß25-35+hepcidin-siRNA+YHG(Aß+siRNA+YHG) group. The expression of ADAM17 mRNA in cells was detected by real-time PCR, and the expression of ADAM17 protein by immunofluorescence and Western blot. Immunofluorescence showed that the ADAM17 protein expression was lower in the Aß group, siRNA group, and Aß+siRNA group than in the control group(P<0.05) and the expression was lower in the Aß+siRNA group(P<0.05) and higher in the Aß+YHG group(P<0.05) than in the Aß group. Moreover, the ADAM17 protein expression was lower in the Aß+siRNA group(P<0.05) and higher in the siRNA+YHG group(P< 0.05) than in the siRNA group. The expression was higher in the Aß+siRNA+YHG group than in the Aß+siRNA group(P<0.05). The results of Western blot and real-time PCR were consistent with those of immunofluorescence. The experiment showed that YHG induced hepcidin to up-regulate the expression of ADAM17 in AD cell model and promote the activation of non-starch metabolic pathways, which might be the internal mechanism of YHG in preventing and treating AD.


Subject(s)
Alzheimer Disease , Drugs, Chinese Herbal , Pueraria , ADAM17 Protein , Alzheimer Disease/drug therapy , Alzheimer Disease/genetics , Amyloid beta-Peptides , Drugs, Chinese Herbal/pharmacology , Hepcidins/genetics , Humans
10.
J Neurosci ; 39(23): 4624-4630, 2019 06 05.
Article in English | MEDLINE | ID: mdl-30926745

ABSTRACT

Temporal lobe epilepsy (TLE) is a common and commonly devastating form of human epilepsy for which only symptomatic therapy is available. One cause of TLE is an episode of de novo prolonged seizures [status epilepticus (SE)]. Understanding the molecular signaling mechanisms by which SE transforms a brain from normal to epileptic may reveal novel targets for preventive and disease-modifying therapies. SE-induced activation of the BDNF receptor tyrosine kinase, TrkB, is one signaling pathway by which SE induces TLE. Although activation of TrkB signaling promotes development of epilepsy in this context, it also reduces SE-induced neuronal death. This led us to hypothesize that distinct signaling pathways downstream of TrkB mediate the desirable (neuroprotective) and undesirable (epileptogenesis) consequences. We subsequently demonstrated that TrkB-mediated activation of phospholipase Cγ1 is required for epileptogenesis. Here we tested the hypothesis that the TrkB-Shc-Akt signaling pathway mediates the neuroprotective consequences of TrkB activation following SE. We studied measures of molecular signaling and cell death in a model of SE in mice of both sexes, including wild-type and TrkBShc/Shc mutant mice in which a point mutation (Y515F) of TrkB prevents the binding of Shc to activated TrkB kinase. Genetic disruption of TrkB-Shc signaling had no effect on severity of SE yet partially inhibited activation of the prosurvival adaptor protein Akt. Importantly, genetic disruption of TrkB-Shc signaling exacerbated hippocampal neuronal death induced by SE. We conclude that therapies targeting TrkB signaling for preventing epilepsy should spare TrkB-Shc-Akt signaling and thereby preserve the neuroprotective benefits.SIGNIFICANCE STATEMENT Temporal lobe epilepsy (TLE) is a common and devastating form of human epilepsy that lacks preventive therapies. Understanding the molecular signaling mechanisms underlying the development of TLE may identify novel therapeutic targets. BDNF signaling thru TrkB receptor tyrosine kinase is one molecular mechanism promoting TLE. We previously discovered that TrkB-mediated activation of phospholipase Cγ1 promotes epileptogenesis. Here we reveal that TrkB-mediated activation of Akt protects against hippocampal neuronal death in vivo following status epilepticus. These findings strengthen the evidence that desirable and undesirable consequences of status epilepticus-induced TrkB activation are mediated by distinct signaling pathways downstream of this receptor. These results provide a strong rationale for a novel therapeutic strategy selectively targeting individual signaling pathways downstream of TrkB for preventing epilepsy.


Subject(s)
Hedgehog Proteins/metabolism , Hippocampus/metabolism , Membrane Glycoproteins/physiology , Nerve Tissue Proteins/physiology , Protein-Tyrosine Kinases/physiology , Sensory Receptor Cells/physiology , Status Epilepticus/metabolism , Animals , Binding Sites , Brain-Derived Neurotrophic Factor/pharmacology , Cell Differentiation/drug effects , Cell Survival/drug effects , Cells, Cultured , Hippocampus/pathology , MAP Kinase Signaling System/drug effects , Mechanotransduction, Cellular , Membrane Glycoproteins/genetics , Membrane Glycoproteins/metabolism , Mice , Mutagenesis, Site-Directed , Nerve Growth Factors/pharmacology , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Phosphorylation , Point Mutation , Protein Binding/genetics , Protein Processing, Post-Translational/drug effects , Protein-Tyrosine Kinases/genetics , Protein-Tyrosine Kinases/metabolism , Recombinant Proteins/metabolism , Sensory Receptor Cells/drug effects , Status Epilepticus/genetics , Touch/physiology
11.
Mar Drugs ; 18(6)2020 Jun 25.
Article in English | MEDLINE | ID: mdl-32630403

ABSTRACT

An efficient strategy was developed for the rapid separation and enrichment of bafilomycin A1 (baf A1) from a crude extract of the marine microorganism Streptomyces lohii fermentation. This strategy comprises liquid-liquid extraction (LLE) with a three-phase solvent system (n-hexane-ethyl acetate-acetonitrile-water = 7:3:5:5, v/v/v/v) followed by separation using high-speed counter-current chromatography (HSCCC). The results showed that a 480.2-mg fraction of baf A1-enriched extract in the middle phase of the three-phase solvent system was prepared from 4.9 g of crude extract after two consecutive one-step operations. Over 99% of soybean oil, the main hydrophobic waste in the crude extract, and the majority of hydrophilic impurities were distributed in the upper and lower phase, respectively. HSCCC was used with a two-phase solvent system composed of n-hexane-acetonitrile-water (15:8:12, v/v/v) to isolate and purify baf A1 from the middle phase fraction, which yielded 77.4 mg of baf A1 with > 95% purity within 90 min. The overall recovery of baf A1 in the process was determined to be 95.7%. The use of a three-phase solvent system represents a novel strategy for the simultaneous removal of hydrophobic oil and hydrophilic impurities from a microbial fermentation extract.


Subject(s)
Countercurrent Distribution/methods , Liquid-Liquid Extraction/methods , Macrolides/isolation & purification , Acetonitriles , Chromatography, High Pressure Liquid , Fermentation , Hexanes , Solvents , Streptomyces
12.
Ecotoxicol Environ Saf ; 188: 109842, 2020 Jan 30.
Article in English | MEDLINE | ID: mdl-31707322

ABSTRACT

Twelve commercial pesticide formulations containing sixteen active ingredients were applied on greenhouse strawberries at recommended and double doses. The dynamics and dietary risk analysis were investigated. A modified QuEChERS method based on the use of multi-walled carbon nanotubes (MWCNTs) as adsorbent followed by LC-MS/MS and GC-MS/MS detection was utilized for sample analysis. The half-lives of studied pesticides were 4.6-12.6 days and 3.8-15.8 days from two application doses. Dietary levels from the residue concentrations of the individual pesticides at harvest was contrasted with the acceptable daily intake (ADI) and acute reference dose (ARfD). The dietary risk assessment adopt the risk quotient (RQ) for chronic risk and risk probability (RP) for long-term dietary intake risk, respectively. The dietary risk induced by the studied pesticide residues in strawberry was acceptable for consumers except the pesticide fumigants. The pesticide residues at different pre-harvest intervals (PHIs) under greenhouse conditions were compared with the established maximum residue limits (MRLs). To reduce the residue levels and potential safety risk, a longer PHI or reduced application rates should be conducted.


Subject(s)
Food Contamination/analysis , Fragaria/chemistry , Fruit/chemistry , Pesticide Residues/analysis , Chromatography, Liquid , Fragaria/growth & development , Fruit/growth & development , Nanotubes, Carbon/chemistry , No-Observed-Adverse-Effect Level , Pesticide Residues/toxicity , Risk Assessment , Tandem Mass Spectrometry
13.
Chem Biodivers ; 17(7): e2000221, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32347603

ABSTRACT

The in situ application of iChip cultivation in mangrove sediment from Hainan province, China, led to the isolation of a novel bacterial species Gallaecimonas mangrovi HK-28. The extract of G. mangrovi HK-28 exhibited antibiotic activity against the aquatic pathogen Vibrio harveyi, and its chemical constituents were further investigated by bioactivity-guided isolation. Three new diketopiperazines, gallaecimonamides A-C, were accordingly isolated from the AcOEt extract of the fermentation broth of G. mangrovi HK-28. The planar structures of gallaecimonamides A-C were determined using HR-ESI-MS together with 1D- and 2D-NMR. The absolute configurations of gallaecimonamides A-C were assigned by optical rotation, NOESY experiment and TDDFT ECD calculations. The in vitro antibacterial and antimalarial activities of gallaecimonamides A-C were assessed. Gallaecimonamide A was found to display antibacterial activity against V. harveyi with a MIC value of 50 µm. However, gallaecimonamides B and C showed no antibacterial activity against V. harveyi (MIC >300 µm). In addition, all the isolates did not exhibit any inhibitory activities against V. parahaemolyticus (MIC>300 µm) and Plasmodium falciparum W2 (EC50 >100 µg/mL).


Subject(s)
Anti-Bacterial Agents/pharmacology , Diketopiperazines/pharmacology , Gammaproteobacteria/chemistry , Vibrio/drug effects , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/isolation & purification , Deuterium , Diketopiperazines/chemistry , Diketopiperazines/isolation & purification , Dose-Response Relationship, Drug , Magnetic Resonance Spectroscopy , Microbial Sensitivity Tests , Molecular Structure , Spectrometry, Mass, Electrospray Ionization , Structure-Activity Relationship
14.
J Infect Dis ; 220(4): 677-686, 2019 07 19.
Article in English | MEDLINE | ID: mdl-31001637

ABSTRACT

Mycobacterium tuberculosis lipid metabolism pathways facilitate access to carbon and energy sources during infection. M. tuberculosis gene Rv1075c was annotated as a conserved hypothetical protein. We identified that Rv1075c amino acid sequence shares similarities with other bacterial lipase/esterases and we demonstrated that it has esterase activity, with preference for short-chain fatty acids, particularly acetate, with highest activity at 45°C, pH 9. Site-direct mutagenesis revealed its activity triad as Ser80, Asp244, and His247. We further determined that rRv1075c hydrolyzed triacetin and tributyrin, and it was mainly distributed in cell wall and membrane. Its expression was induced at pH 4.5, mimicking the acidic phagosome of macrophages. Mutation of Rv1075c led to reduced bacterial growth in THP-1 cells and human peripheral blood mononuclear cell-derived macrophages, and attenuated M. tuberculosis infection in mice. Our data suggest that Rv1075c is involved in ester and fatty acid metabolism inside host cells.


Subject(s)
Bacterial Proteins/metabolism , Esterases/metabolism , Lipid Metabolism , Mycobacterium tuberculosis/enzymology , Tuberculosis/microbiology , Amino Acid Sequence , Animals , Bacterial Proteins/genetics , Cell Wall/enzymology , Cytoplasm/metabolism , Esterases/genetics , Female , Humans , Hydrogen-Ion Concentration , Hydrolysis , Leukocytes, Mononuclear , Lipase/genetics , Lipase/metabolism , Macrophages/microbiology , Mice , Models, Structural , Mutagenesis, Site-Directed , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/physiology , Sequence Alignment , THP-1 Cells
15.
EMBO J ; 34(5): 609-23, 2015 Mar 04.
Article in English | MEDLINE | ID: mdl-25603933

ABSTRACT

Enhanced glycolysis is a main feature of pluripotent stem cells (PSCs) and is proposed to be important for the maintenance and induction of pluripotency. The molecular mechanism underlying enhanced glycolysis in PSCs is not clear. Using Dgcr8-/- mouse embryonic stem cells (ESCs) that lack mature miRNAs, we found that miR-290 cluster of miRNAs stimulates glycolysis by upregulating glycolytic enzymes Pkm2 and Ldha, which are also essential for the induction of pluripotency during reprogramming. Mechanistically, we identified Mbd2, a reader for methylated CpGs, as the target of miR-290 cluster that represses glycolysis and reprogramming. Furthermore, we discovered Myc as a key target of Mbd2 that controls metabolic switch in ESCs. Importantly, we demonstrated that miR-371 cluster, a human homolog of miR-290 cluster, stimulates glycolysis to promote the reprogramming of human fibroblasts. Hence, we identified a previously unappreciated mechanism by which miR-290/371 miRNAs orchestrate epigenetic, transcriptional and metabolic networks to promote pluripotency in PSCs and during reprogramming.


Subject(s)
DNA-Binding Proteins/metabolism , Glycolysis/physiology , Metabolic Networks and Pathways/physiology , MicroRNAs/metabolism , Pluripotent Stem Cells/enzymology , Pluripotent Stem Cells/physiology , Proto-Oncogene Proteins c-myc/metabolism , Animals , Chromatin Immunoprecipitation , DNA Primers/genetics , DNA-Binding Proteins/genetics , Gene Knockout Techniques , Glycolysis/genetics , Metabolic Networks and Pathways/genetics , Mice , Mice, Knockout , MicroRNAs/genetics , Proto-Oncogene Proteins c-myc/genetics , RNA, Small Interfering/genetics , RNA-Binding Proteins/genetics , RNA-Binding Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction
16.
Mol Ecol ; 28(5): 1138-1153, 2019 03.
Article in English | MEDLINE | ID: mdl-30614581

ABSTRACT

Current monitoring methods to assess benthic impacts of marine finfish aquaculture are based on complex biological indices and/or geochemistry data. The former requires benthic macrofauna morpho-taxonomic characterization that is time- and cost-intensive, while the latter provides rapid assessment of the organic enrichment status of sediments but does not directly measure biotic impacts. In this study, sediment samples were collected from seven stations at six salmon farms in British Columbia, Canada, and analyzed for geochemical parameters and by eDNA metabarcoding to investigate linkages between geochemistry and foraminifera. Sediment texture across farm sites ranged from sand to silty loam, while the maximum sediment pore-water sulphide concentration at each site ranged from 1,000 to 13,000 µM. Foraminifera alpha diversity generally increased with distance from cage edge. Adonis analyses revealed that farm site explained the most variation in foraminifera community, followed by sediment type, enrichment status, and distance from cage edge. Farm-specific responses were observed in diversity analyses, taxonomic difference analyses, and correlation analyses. Results demonstrated that species diversity and composition of foraminifera characterized by eDNA metabarcoding generated signals consistent with benthic biodiversity being impacted by finfish farming activities. This substantiates the validity of eDNA metabarcoding for augmenting current approaches to benthic impact assessments by providing more cost-effective and practicable biotic measures than traditional morpho-taxonomy. To capitalize on this potential, further work is needed to design a new nomogram that combines eDNA metabarcoding data and geochemistry data to enable accurate monitoring of benthic impacts of fish farming in a time- and cost-efficient way.


Subject(s)
DNA Barcoding, Taxonomic , DNA/genetics , Foraminifera/genetics , Salmon/genetics , Animals , Aquaculture , Biodiversity , British Columbia , Environmental Monitoring , Fisheries , Geologic Sediments/chemistry , Salmon/growth & development
17.
EMBO Rep ; 18(5): 809-825, 2017 05.
Article in English | MEDLINE | ID: mdl-28325773

ABSTRACT

Cancer progression depends on cellular metabolic reprogramming as both direct and indirect consequence of oncogenic lesions; however, the underlying mechanisms are still poorly understood. Here, we report that CUEDC2 (CUE domain-containing protein 2) plays a vital role in facilitating aerobic glycolysis, or Warburg effect, in cancer cells. Mechanistically, we show that CUEDC2 upregulates the two key glycolytic proteins GLUT3 and LDHA via interacting with the glucocorticoid receptor (GR) or 14-3-3ζ, respectively. We further demonstrate that enhanced aerobic glycolysis is essential for the role of CUEDC2 to drive cancer progression. Moreover, using tissue microarray analysis, we show a correlation between the aberrant expression of CUEDC2, and GLUT3 and LDHA in clinical HCC samples, further demonstrating a link between CUEDC2 and the Warburg effect during cancer development. Taken together, our findings reveal a previously unappreciated function of CUEDC2 in cancer cell metabolism and tumorigenesis, illustrating how close oncogenic lesions are intertwined with metabolic alterations promoting cancer progression.


Subject(s)
Carcinogenesis , Carrier Proteins/metabolism , Glycolysis , Membrane Proteins/metabolism , 14-3-3 Proteins/metabolism , Adaptor Proteins, Signal Transducing , Carrier Proteins/genetics , Cell Line, Tumor , Disease Progression , Gene Expression Regulation, Neoplastic , Glucose Transporter Type 3/genetics , Glucose Transporter Type 3/metabolism , HeLa Cells , Humans , Isoenzymes/genetics , Isoenzymes/metabolism , L-Lactate Dehydrogenase/genetics , L-Lactate Dehydrogenase/metabolism , Lactate Dehydrogenase 5 , Membrane Proteins/genetics , Receptors, Glucocorticoid/metabolism , Tissue Array Analysis , Transcriptional Activation , Up-Regulation
18.
J Sep Sci ; 42(15): 2510-2516, 2019 Aug.
Article in English | MEDLINE | ID: mdl-31127863

ABSTRACT

High-speed counter-current chromatography was applied to the separation of five diketoperazines from the marine Alternaria alternate HK-25 for the first time using one-step elution method with a pair of two-phase solvent systems composed of petroleum ether/ethyl acetate/methanol/water (5.5:11:5:7, v/v). Where 151.6 mg of crude sample yielded five diketoperazines, 12,13-dihydroxy-fumitremorgin C (1), gliotoxin (2), demethoxyfum itremorgin C (3), bisdethiobis(methylthio)gliotoxin (4), fumitremorgin C (5), and the purities of all compounds were above 94% as determined by high-performance liquid chromatography. The structures of these compounds were identified by 1 H and 13 C NMR spectroscopy. These results showed that high-speed counter-current chromatography can provide a feasible way for highly effective preparation of marine natural products, which ensured the supple of numerous samples for drug development.


Subject(s)
Alternaria/chemistry , Biological Products/isolation & purification , Diketopiperazines/isolation & purification , Biological Products/chemistry , Countercurrent Distribution , Diketopiperazines/chemistry , Molecular Conformation
19.
J Environ Sci (China) ; 77: 282-290, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30573092

ABSTRACT

Iron ore sintering is a major source of gaseous and particulate pollutants emission in iron smelt plant. The aim of present study is to characterize the volatile organic compounds (VOCs) emission profiles from iron ore sintering process. Both sinter pot test and sinter simulation experiment were conducted and compared. Out results showed that sinter process produced large quantity of VOCs together with NOx and SO2. VOCs and NO were produced simultaneously in sinter pot test from 3 to 24 min after ignition, flowed by SO2 production from 15 min to the end of sintering. Total VOCs (TVOC) concentration in sinter flue gas was affected by the coal and coke ratio in sinter raw material. The maximum TVOC concentration was 34.5 ppm when using 100% coal as fuel. Sinter simulation experiments found that the number of VOCs species and their concentrations were found by sinter temperature. The largest VOCs species varieties were obtained at 500 °C. Benzene, toluene, xylene and ethylbenzene were major VOCs in sinter flue gas based on the results from both simulation test and sinter pot. It thus demonstrated that in addition to NOx, SO2 and metal oxide particles, sinter flue gas also contained significant amount of VOCs whose environmental impact cannot be ignored. Based on our work, it is timely needed to establish a new VOC emission standard for sinter flue gas and develop advanced techniques to simultaneously eliminate multi-pollutants in iron ore sinter process.


Subject(s)
Iron/chemistry , Volatile Organic Compounds/chemistry , Hot Temperature , Oxides/chemistry , Particulate Matter/chemistry
20.
Microb Ecol ; 75(1): 140-151, 2018 Jan.
Article in English | MEDLINE | ID: mdl-28714057

ABSTRACT

The gut microbial community in vertebrates plays a role in nutrient digestion and absorption, development of intestine and immune systems, resistance to infection, regulation of bone mass and even host behavior and can thus impact host fitness. Atlantic salmon (Salmo salar) reintroduction efforts into Lake Ontario, Canada, have been unsuccessful, likely due to competition with non-native salmonids. In this study, we explored interspecific competition effects on the gut microbiota of two Atlantic salmon populations (LaHave and Sebago) resulting from four non-native salmonids. After 10 months of rearing in semi-natural stream tanks under six interspecific competition treatments, we characterized the gut microbiota of 178 Atlantic salmon by parallel sequencing the 16S rRNA gene. We found 3978 bacterial OTUs across all samples. Microbiota alpha diversity and abundance of 27 OTUs significantly differed between the two populations. Interspecific competition reduced relative abundance of potential beneficial bacteria (six genera of lactic acid bacteria) as well as 13 OTUs, but only in the LaHave population, indicating population-specific competition effects. The pattern of gut microbiota response to interspecific competition may reflect local adaptation of the host-microbiota interactions and can be used to select candidate populations for improved species reintroduction success.


Subject(s)
Bacterial Physiological Phenomena , Gastrointestinal Microbiome , Intestines/microbiology , Salmo salar/microbiology , Animals , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , DNA, Bacterial/genetics , Ontario , RNA, Ribosomal, 16S/genetics , Salmo salar/classification
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