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1.
Mitochondrial DNA B Resour ; 9(5): 616-620, 2024.
Article in English | MEDLINE | ID: mdl-38737393

ABSTRACT

Tylopilus brunneirubens is a common species in southern China. It is known for brown to dark brown pileus, white context turning reddish brown or rust brown when touched and distinct reticulation on the upper stem. However, little is known about its mitochondrial genome and its relationship with other boletes. Our analysis revealed that the mitochondrial genome of this species is a circular DNA molecule that spans 32,389 bp. It contains 15 core protein-coding genes, 24 transfer RNA genes, and two ribosomal RNA genes. The base composition of the mitochondrial genome is as follows: A (37.20%), C (11.32%), G (12.48%), and T (39.00%), with a GC content of 23.80%. Furthermore, a phylogenetic tree based on 24 mitochondrial genomes provided valuable insights into the phylogenetic relationships of Tylopilus brunneirubens with other boletes for the first time.

2.
Mitochondrial DNA B Resour ; 9(1): 46-49, 2024.
Article in English | MEDLINE | ID: mdl-38197052

ABSTRACT

Butyriboletus hainanensis, a macrofungus belonging to the Boletaceae family, is named after its collection location on Hainan Island, China. However, little is known about its mitochondrial genome and its phylogenetic relationship with other boletes. In this study, we utilized next-generation sequencing technology to sequence the mitochondrial genome of Bu. hainanensis. Our findings revealed that the mitochondrial genome of this species is presumably a circular DNA molecule spanning 36,592 bp. It consists of 15 protein-coding genes, 27 transfer RNA genes, and two ribosomal RNA genes. The base composition of the mitochondrial genome is as follows: A (36.64%), C (12.22%), G (11.73%), and T (39.41%), with a GC content of 23.95%. Additionally, a phylogenetic tree was constructed based on 22 mitochondrial genomes, which provided valuable insights into the phylogenetic relationships of Bu. hainanensis with other boletes for the first time.

3.
Front Microbiol ; 13: 1070773, 2022.
Article in English | MEDLINE | ID: mdl-36532431

ABSTRACT

Over the past few decades, there has been a noticeable surge in the market of plant-based meat analogs (PBMA). Such popularity stems from their environmentally friendly production procedures as well as their positive health effects. In order to meet the market demand, it is necessary to look for plant protein processing techniques that can help them match the quality of conventional meat protein from the aspects of sensory, quality and functionality. Bean proteins are ideal options for PBMA with their easy accessibility, high nutrient-density and reasonable price. However, the high polyunsaturated lipids content of beans inevitably leads to the unpleasant beany flavor of soy protein products, which severely affects the promotion of soy protein-based PBMA. In order to solve this issue, various methods including bleaching, enzyme and fermentation etc. are developed. Among these, fermentation is widely investigated due to its high efficiency, less harm to the protein matrix, targeted performance and low budget. In addition, proper utilization of microbiome during the fermentation process not only reduces the unpleasant beany flavors, but also enhances the aroma profile of the final product. In this review, we provide a thorough and succinct overview of the mechanism underlying the formation and elimination of beany flavor with associated fermentation process. The pros and cons of typical fermentation technologies for removing beany flavors are discussed in alongside with their application scenarios. Additionally, the variations among different methods are compared in terms of the strains, fermentation condition, target functionality, matrix for application, sensory perception etc.

4.
Comput Intell Neurosci ; 2022: 4051955, 2022.
Article in English | MEDLINE | ID: mdl-36059410

ABSTRACT

Methods: From April 2020 to May 2021, 56 patients with CHF who were discharged from the cardiology department of our hospital after treatment were randomly divided into two groups: experimental group (n = 28) and control group (n = 28). The control group was given conventional nursing measures and health education and discharge instructions, while the experimental group received collaborative multidisciplinary team nursing care based on the WeChat platform on the basis of the control group, all for 3 months. All enrolled patients underwent the Self-Care of Heart Failure Index Version 6.2 (SCHFI v6.2), the Minnesota Living with Heart Failure Questionnaire (MLHFQ), and the 6-minute walking test (6MWT test). The SCHFI v6.2 and MLHFQ scores, 6 MWT test results, and readmission rates within 3 months were observed and compared between the two groups. Results: There was no significant difference between the SCHFIv6.2 and MLHFQ scores of the two patients at admission and at discharge, and the scores of the experimental group were significantly higher than the scores of the control group at the end of 3 months after discharge. On the other hand, the SCHFIv6.2 and MLHFQ scores of the two groups were significantly higher at discharge compared to admission; the 6-minute walking distance of the experimental group was significantly higher than that of the control group at the end of 3 months. The readmission rate in the experimental group was significantly lower than that in the control group. Conclusion: The multidisciplinary teamwork model based on the WeChat platform can significantly improve the self-care ability and quality of life of CHF patients and reduce the readmission rate.


Subject(s)
Heart Failure , Quality of Life , Heart Failure/diagnosis , Heart Failure/therapy , Humans , Patient Care Team , Surveys and Questionnaires
5.
ACS Appl Mater Interfaces ; 13(21): 24774-24784, 2021 Jun 02.
Article in English | MEDLINE | ID: mdl-34015919

ABSTRACT

With the rapid development in wearable electronics, self-powered devices have recently attracted tremendous attention to overcome the restriction of conventional power sources. In this regard, a simple, scalable, and one-pot electrospinning fabrication technique was utilized to construct an all-fiber-structured triboelectric nanogenerator (TENG). Ethyl cellulose was co-electrospun with polyamide 6 to serve as the triboelectric positive material, and a kind of strongly electronegative conductive material of MXene sheet was innovatively incorporated into poly(vinylidene fluoride) nanofiber to act as a triboelectric negative material. The assembled all-fiber TENG exhibited excellent durability and stability, as well as excellent output performance, which reached a peak power density of 290 mW/m2 at a load resistance of 100 MΩ. More importantly, the TENG was capable of harvesting energy to power various light-emitting diodes (LEDs) and monitoring human movements as a self-powered sensor, providing a promising application prospect in wearable electronics.

6.
Nanomaterials (Basel) ; 11(2)2021 Feb 03.
Article in English | MEDLINE | ID: mdl-33546254

ABSTRACT

A "super" bioactive antibacterial hydrogel, Icariin-ß-CD-inclusion complex/Bacterial cellulose and an equally capable counterpart Icariin-Bacterial cellulose (ICBC) were successfully produced with excellent antioxidant properties. The highly porous hydrogels demonstrated very high fluid/liquid absorption capability and were functionally active as Fourier Transform Infrared Spectrometer (FTIR) test confirmed the existence of abundant hydroxyls (-OH stretching), carboxylic acids (-CH2/C-O stretching), Alkyne/nitrile (C≡C/C≡N stretching with triple bonds) and phenol (C-H/N-O symmetric stretching) functional groups. Scanning electron microscope (SEM) and X-ray diffraction (XRD) tests confirmed a successful ß-CD-inclusion complexation with Icariin with a great potential for sustained and controlled drug release. In vitro drug release test results indicated a systemic and controlled release of the drug (Icariin) from the internal cavities of the ß-CD inclusion complex incorporated inside the BC matrix with high Icariin (drug) release rates. Impressive inactivation rates against Gram-negative bacteria Escherichia coli ATCC 8099 and gram-positive bacteria Staphylococcus aureus ATCC 6538; >99.19% and >98.89% respectively were recorded, as the materials proved to be non-toxic on L929 cells in the in vitro cytotoxicity test results. The materials with promising versatile multipurpose administration of Icariin for wound dressing (as wound dressers), can also be executed as implants for tissue regeneration, as well as face-mask for cosmetic purposes.

7.
Biosens Bioelectron ; 171: 112690, 2021 Jan 01.
Article in English | MEDLINE | ID: mdl-33049561

ABSTRACT

Development of reliable Enzymatic Biofuel Cells (EBFC)-based self-powered glucose biosensor for continuous, noninvasive monitoring without restriction on patient's movement is highly recommendable. However, its application to a large extent is limited by the relatively poor stability. Herein, we synthesized a highly flexible electrode for effective enzyme immobilization by encapsulating enzyme into the metal-organic frameworks (MOFs) and robustly anchored to the cellulose acetate (CA) nanofiber membrane. As is well-known, such nanostructured fiber materials are the first time to be synthesized for glucose biosensor, which encapsulated biomolecules in MOFs platform during the MOFs in-situ growth on the nanofiber membranes. The as-proposed biosensor demonstrated excellent stability over 15 h of continuous long-term monitoring. The remarkable stability of assembled self-powered glucose biosensor in this work could inspire the application of enzymatic biosensors in biometrics, chronic disease management and clinical diagnosis.


Subject(s)
Biosensing Techniques , Glucose , Metal-Organic Frameworks , Nanofibers , Cellulose/analogs & derivatives , Glucose/analysis , Humans
8.
J Pediatr ; 216: 204-207, 2020 01.
Article in English | MEDLINE | ID: mdl-31732130

ABSTRACT

OBJECTIVE: To test, in a newborn screening (NBS) laboratory, the performance of liquid chromatography-tandem mass spectrometry (LC-MS/MS) to assay 5 enzymatic activities in dried blood spots (DBS) for NBS of 5 lysosomal storage diseases (mucopolysaccharidosis [MPS]-II, MPS-IIIB, MPS-IVA, MPS-VI, and MPS-VII). STUDY DESIGN: Three mm punches from de-identified DBS were obtained from the Washington NBS laboratory and submitted to the 5-plex LC-MS/MS assay. Screen cut-offs were established by analyzing the enzymatic activity in patients confirmed to have the MPS disorder. DNA sequencing of the relevant gene was performed on a second DBS punch for all samples with enzyme activity below 10% of the mean daily activity. RESULTS: (1) For MPS-II, 18 below cut-off samples, 1 pathogenic genotype, and 2 "high risk" genotypes; (2) For MPS-IIIB, no below cut-off samples; (3) For MPS-IVA, 8 below cut-off samples, 4 non-pathogenic genotypes, 4 genotypes unobtainable; (4) For MPS-VI, 4 below cut-off samples and no high-risk genotypes; (5) For MPS-VII, 1 below cut-off sample confirmed by genotype and clinical report to be affected. CONCLUSIONS: These results establish that the number of initial screen positive samples is low and manageable. Thus, population newborn screening for these conditions is feasible in a state newborn screening laboratory.


Subject(s)
Mucopolysaccharidoses/blood , Mucopolysaccharidoses/diagnosis , Neonatal Screening , Chromatography, Liquid , Dried Blood Spot Testing/statistics & numerical data , Humans , Infant, Newborn , Mucopolysaccharidoses/enzymology , Pilot Projects , Tandem Mass Spectrometry
9.
Anticancer Agents Med Chem ; 20(1): 94-102, 2020.
Article in English | MEDLINE | ID: mdl-31746308

ABSTRACT

BACKGROUND AND PURPOSE: Although limited by side effects and development of resistance, doxorubicin still represents the most common chemotherapy for breast cancer. Thus, the identification of critical molecules to alleviate doxorubicin resistance is crucial. Here, we provide a molecular rationale for the breast cancer patients potentially benefitting from doxorubicin based on the expression levels of SIRT1, an identified member of longevity genes. METHODS: SIRT1-overexpressed and SIRT1-knockdown breast cancer cells were established to investigate the functions of SIRT1 in regulating doxorubicin resistance both in vitro and in vivo. Cell proliferation was analyzed via CCK8 assay, cell apoptosis was studied by TUNEL analysis. Molecule interaction was analyzed through co-immunoprecipitation and immunofluorescence techniques. Sensibility to doxorubicin was assessed in vivo through the nude mice tumorigenicity experiment. RESULTS: First, SIRT1 was found higher-expressed in breast cancer doxorubicin-resistant cells MCF-7/ADR than that in the doxorubicin- sensitive cells MCF-7. Moreover, SIRT1-knockdown MCF-7/ADR cells showed higher susceptible to doxorubicin both in vitro and in vivo models, whereas overexpressing of SIRT1 inhibited this phenotype. Accordingly, SIRT1 was found interacted with Akt, consequently promoted the activity of Akt in MCF-7/ADR cells in vitro and positively correlated with the expression of P-Akt in vivo. Reversing the activity of Akt partially downturned the doxorubicin-resistant effects mediated by SIRT1. CONCLUSION: This investigation suggested the value of SIRT1 as a biomarker of response to doxorubicin, leading to the development of new tools for the management of breast cancer patients.


Subject(s)
Antibiotics, Antineoplastic/pharmacology , Breast Neoplasms/drug therapy , Doxorubicin/pharmacology , Drug Resistance, Neoplasm/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Sirtuin 1/genetics , Antibiotics, Antineoplastic/chemistry , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Dose-Response Relationship, Drug , Doxorubicin/chemistry , Drug Screening Assays, Antitumor , Female , Humans , Structure-Activity Relationship
10.
Pathol Res Pract ; 215(12): 152702, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31704152

ABSTRACT

The ETFDH (electron transfer flavoprotein dehydrogenase) gene mutations are reported to be a major cause of riboflavin-responsive multiple acyl-coenzyme A dehydrogenation deficiency (MADD). However, the role of ETFDH in the prognosis of hepatocellular carcinoma (HCC) remains unclear. The aim of this study was to investigate the expression of ETFDH in HCC. Immunohistochemical staining of the 207 HCC tissue microarray showed that expression of ETFDH was significantly decreased in HCC compared with the matching noncancerous hepatic tissues (P < 0.001). Moreover, ETFDH expression levels were found to be correlated with AFP levels (P = 0.011). Intriguingly, ETFDH expression levels were significantly lower in poorly differentiated or undifferentiated HCCs as compared to the well or moderately differentiated cases (P = 0.001). Kaplan-Meier analysis revealed that low tumor expression of ETFDH was associated with a poorer overall survival in patients with HCC (P = 0.024). Furthermore, multivariate analysis showed that ETFDH (P = 0.047) was an independent predictor of overall survival. Our findings may shed new light on the identification of new prognostic marker for HCC.


Subject(s)
Biomarkers, Tumor/analysis , Carcinoma, Hepatocellular/enzymology , Electron-Transferring Flavoproteins/analysis , Iron-Sulfur Proteins/analysis , Liver Neoplasms/enzymology , Oxidoreductases Acting on CH-NH Group Donors/analysis , Adult , Aged , Carcinoma, Hepatocellular/mortality , Carcinoma, Hepatocellular/pathology , Carcinoma, Hepatocellular/therapy , Cell Differentiation , Female , Humans , Immunohistochemistry , Liver Neoplasms/mortality , Liver Neoplasms/pathology , Liver Neoplasms/therapy , Male , Middle Aged , Prognosis , Risk Factors , Tissue Array Analysis , alpha-Fetoproteins/analysis
11.
ChemSusChem ; 12(23): 5075-5080, 2019 Dec 06.
Article in English | MEDLINE | ID: mdl-31609527

ABSTRACT

To protect the structure of MoS2 from collapse, a strong skeleton is expected to help maintain the integrity. In this study, cotton linters burdened with hollow C@MoS2 nanocapsules are added into nutrient medium for the growth of a bacterial cellulose membrane. Benefitting from good conductivity and structural integrity, the resultant fibrous membrane anode gives reversible capacities of 559 and 155 mAh g-1 for Li-ion batteries and Na-ion batteries after 100 cycles, respectively. The structural transformation and component evolution in lithiation-delithiation and sodiation-desodiation was elucidated by in situ Raman spectroscopy. After sodiation, the Na2 S did not transform back into MoS2 but was more likely converted into elemental sulfur during the conversion reaction. Layered semiconducting transition metal chalcogenides, such as molybdenum disulfide (MoS2 ), feature open 2 D ion-transport channels amenable to receive various guest ions with high theoretical capacities.[2] One serious challenge curtailing the applicability of such materials is their volume changes during discharge-charge processes.[3, 4] However, particular morphologies of MoS2 are proposed to improve the specific capacity.[5,6,7] Many works have focused on core-shell and hollow MoS2 micro- and nanostructures, and the results validate the advantages of shortening the lithium-ion diffusion distance and enhancing specific capacity.[8,9] Unfortunately, the issue of inferior capacity stability is not resolved, because the structure is not effectively protected and is prone to collapse.

12.
Pathol Res Pract ; 215(11): 152603, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31540771

ABSTRACT

miR-182 is a well-described oncogenic miRNA playing a crucial role in the development of many malignancies. However, the role of miR-182 in multiple myeloma (MM) remains unclear. Here, we demonstrate that adhesion of H929 and MM.1S cells to fibronectin could induce miR-182 expression and decrease PDCD4 expression. Furthermore, miR-182 was found to negatively regulate PDCD4 expression in H929 and MM.1S cells. In addition, PDCD4 down-regulation was required for cell adhesion-mediated drug resistance (CAM-DR). Intriguingly, miR-182 up-regulation could promote CAM-DR in H929 and MM.1S cells. Moreover, miR-182 up-regulation and PDCD4 down-regulation enhanced AKT phosphorylation at Ser473 in both H929 and MM.1S cells. Our data suggest that cell adhesion-mediated miR-182 up-regulation and PDCD4 down-regulation may confer drug resistance via enhancing AKT phosphorylation at Ser473.


Subject(s)
Apoptosis Regulatory Proteins/biosynthesis , Drug Resistance, Neoplasm/genetics , Gene Expression Regulation, Neoplastic/genetics , MicroRNAs/genetics , Multiple Myeloma/pathology , RNA-Binding Proteins/biosynthesis , Cell Adhesion/drug effects , Cell Line, Tumor , Humans , Multiple Myeloma/genetics , Proto-Oncogene Proteins c-akt/biosynthesis
13.
Pathol Res Pract ; 215(3): 478-482, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30611622

ABSTRACT

PDH E1 component subunit alpha (PDHA1) has been reported to be biologically significant in several human tumors. The aim of this study was to investigate the expression of PDHA1 in gastric cancer (GC) and its relationship with clinicopathological characteristics and prognosis. Oncomine analysis of neoplastic vs. normal tissue showed that the mRNA levels of PDHA1 were significantly underexpressed in different types of GC across three analyses. Underexpression of PDHA1 was found in intestinal-type GC (P = 0.009), diffuse-type GC (P = 0.036), and mixed-type GC (P = 0.025). Immunohistochemical staining of the 174 GC tissue microarray showed that PDHA1 staining is much stronger in normal mucosa than in GC samples (P = 0.040). Furthermore, PDHA1 expression levels were found to be significantly lower in 69.05% (87/126) of poorly differentiated GCs as compared to the well or moderately differentiated ones (P = 0.037). Intriguingly, PDHA1 expression was significantly correlated with depth of invasion (P < 0.001), lymph node metastasis (P < 0.001), TNM stage (P < 0.001), and nerve invasion (P = 0.006). However, it was not correlated with gender, age, Lauren classification, and lymphovascular invasion (P > 0.05 for all). Kaplan-Meier analysis revealed that low tumor expression of PDHA1 was significantly correlated with a poorer overall survival in patients with GC (5-year overall survival rates for patients with low vs high PDHA1 expression = 49.8% vs 72.7%, hazard ratio of death from GC = 2.594, 95% CI = 1.527 to 4.408, P < 0.001). Multivariate analysis showed that PDHA1 (P = 0.025) was an independent predictor of overall survival. These findings are of potential clinical utility and merit further validation.


Subject(s)
Adenocarcinoma/pathology , Biomarkers, Tumor/analysis , Pyruvate Dehydrogenase (Lipoamide)/biosynthesis , Stomach Neoplasms/pathology , Adenocarcinoma/mortality , Adult , Aged , Aged, 80 and over , Female , Humans , Kaplan-Meier Estimate , Male , Middle Aged , Prognosis , Stomach Neoplasms/mortality , Young Adult
14.
Materials (Basel) ; 12(2)2019 Jan 10.
Article in English | MEDLINE | ID: mdl-30634615

ABSTRACT

In this study, a novel green adsorbent material prepared by the esterification of bacterial cellulose (BC) and graphene oxide (GO), richly containing hydroxyl, alkyl, and carboxylate groups was characterised by FTIR (Fourier Transform infrared spectroscopy), XRD (X-ray diffraction), SEM (Scanning electron microscopy) and TGA (Thermo-graphimetric analysis). The specific surface area (SSA) and pore size distribution (PSD) analysis of materials were also analysed. Batch experiments⁻adsorption studies confirmed the material to have a very high Pb2+ removal efficiency of over 90% at pH 6⁻8. Kinetic studies showed that the uptake of metal ions was rapid with equilibrium attained after 30 min and fitted well with the pseudo-second-order rate model (PSO). Isotherm results with a maximum adsorption capacity (Qmax) of 303.03 mg/g were well described by Langmuir's model compared to Freundlich. Desorption and re-adsorption experiments realised that both adsorbent and adsorbates could be over 90⁻95% efficiently recovered and reused using 0.1 M HNO3 and 0.1 M HCl.

15.
Carbohydr Polym ; 172: 93-101, 2017 Sep 15.
Article in English | MEDLINE | ID: mdl-28606552

ABSTRACT

Nitrogen-doped carbon dots (N-CDs) were synthesized through a facile hydrothermal method using citric acid (CA) and ethanediamine (EDA) as precursors. A green and simple fluorescence biosensor was obtained by biosynthesis of bacterial cellulose (BC)/N-CDs. As-prepared N-CDs with rich functional groups exhibited a blue emission under the excitation wavelength of 350nm. Biosynthesis of BC/N-CDs was analyzed by Digital photos, Fourier Transform Infrared (FTIR) and Transmission Electron Microscopy (TEM). The results indicated that N-CDs were successfully anchored on BC. As-prepared BC/N-CDs were applied to the detection of Fe3+ in aqueous solution. Spectroscopic data revealed that, fluorescence materials prepared presented an sensitive response to Fe3+ in acceptable range of 0.5-600µM and ultralow detection limit of 84nM as a fluorescence sensor. Furthermore, the results also indicate that a novel BC/N-CDs composite has great potential for the detection of Fe3+ ions based on membrane fluorescence materials.


Subject(s)
Carbon/chemistry , Cellulose/chemistry , Iron/analysis , Nitrogen , Quantum Dots
16.
Tumour Biol ; 37(1): 1369-78, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26298725

ABSTRACT

Recent studies have identified that thyroid hormone receptor-interacting protein 6 (TRIP6) is implicated in tumorigenesis. However, the functional role of TRIP6 in non-Hodgkin's lymphoma (NHL) has never been elucidated. In this study, we demonstrated that TRIP6 is reversely correlated with the clinical outcomes of NHL patients. Western blot and immunohistochemical analysis revealed that TRIP6 expression is lower in indolent lymphoma than in progressive lymphoma. Kaplan-Meier survival curves indicated that the upregulation of TRIP6 is significantly associated with poor overall survival. Moreover, patients with higher expression of TRIP6 are prone to shorter time to recurrence. Furthermore, we also found that TRIP6 can promote the proliferation of NHL cells via regulating cell cycle progression. In addition, adhesion of lymphoma cells to fibronectin (FN) decreased TRIP6 expression, which led to the upregulation of nuclear p27(Kip1) expression by decreasing phosphorylation of p27(Kip1) at T157. Importantly, overexpression of TRIP6 can reverse cell adhesion-mediated drug resistance (CAM-DR) phenotype in NHL. In summary, these results suggest that TRIP6 is a novel prognostic indicator for NHL patients and may shed new insights into the important role of TRIP6 in cancer development.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Drug Resistance, Neoplasm , Gene Expression Regulation, Neoplastic , LIM Domain Proteins/metabolism , Lymphoma, Non-Hodgkin/metabolism , Transcription Factors/metabolism , ATPases Associated with Diverse Cellular Activities , Aged , Cell Adhesion , Cell Cycle , Cell Proliferation , Cell Survival , Female , Flow Cytometry , Humans , Kaplan-Meier Estimate , Male , Middle Aged , Neoplasm Recurrence, Local , Phenotype , Prognosis , Proteasome Endopeptidase Complex , Treatment Outcome
19.
Protein J ; 31(1): 75-83, 2012 Jan.
Article in English | MEDLINE | ID: mdl-22160327

ABSTRACT

A peroxisome proliferator-actived receptor (PPAR) response element (RE) in the promoter region of the adaptor-related protein complex 2, alpha 2 subunit (AP2α2) of mouse heart has been identified. The steroid hormone nuclear PPARs and the retinoid X receptors (RXRs) are important transcriptional factors that regulate gene expression, cell differentiation and lipid metabolism. They form homo- (RXR) and hetero- (PPAR-RXR) dimers that bind DNA at various REs. The AP2α2 gene is part of complex and process that transports lipids and proteins from the plasma membrane to the endosomal system. A PPAR activator (Wy14643) and DMSO (vehicle) was introduced into control and δ337T thyroid hormone receptor (TRß1) transgenic mice. Heart tissue was extracted and AP2α2 gene expression was compared using Affymetrix expression arrays and qRT PCR among four groups [control, control with Wy14643, δ337T TRß1 and δ337T TRß1 with Wy14643]. The gene expression of AP2α2 in the Wy14643 control and transgenic mouse groups was significantly up regulated over the vehicle mouse groups in both the array (p < 0.01) and qRT PCR (p < 0.01) studies. Duplex oligo DNAs containing the PPAR/RXR motif (AGGTCA/TCCAGT) from the AP2α2 promoter were used in EMSA to verify binding of the PPAR and RXR receptors to their REs. pGL4.0 [Luc] constructs of the AP2α2 promoter with and without the PPAR/RXR motifs were co-transfected with mouse PPARα, ß or γ1 into HepG2 cells and used in lucerifase assays to verify gene activation. In conclusion our study revealed that PPARα regulates the mouse cardiac AP2α2 gene in both the control and transgenic mouse.


Subject(s)
Adaptor Protein Complex 2/genetics , Adaptor Protein Complex alpha Subunits/genetics , Myocardium/metabolism , PPAR alpha/metabolism , Up-Regulation , Adaptor Protein Complex 2/metabolism , Adaptor Protein Complex alpha Subunits/metabolism , Animals , Cell Line , Gene Expression Regulation , Humans , Mice , Mice, Transgenic , PPAR alpha/genetics , Promoter Regions, Genetic , Response Elements
20.
FEBS J ; 276(12): 3247-55, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19438714

ABSTRACT

The purpose of this study was to provide a better understanding of the regulatory role of the nuclear steroid receptor on the nuclear factor of kappa light polypeptide gene enhancer in B cells (NFkappaB) in mouse heart. NFkappaB regulates many nuclear genes and has been associated with many human cardiac diseases. NFkappaB's protein regulator gene, nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor alpha gene (IkappaBalpha), was found in this study to be regulated by peroxisome proliferator-activated receptors (PPARs). PPARs, retinoid X receptors (RXRs) and thyroid hormone receptors (THRs) are members of the nuclear receptor superfamily, which consists of a large number of transcription factors whose activities are regulated by their cognate ligands. These steroid hormone receptors are important regulators of gene expression and differentiation in the heart. These receptors form homo-(RXR, THR) and hetero-(PPAR-RXR, RXR-THR) dimers that bind DNA at various response elements (PPAR, RXR and THR) in the promoter regions of target genes. The PPAR/RXR response elements in the promoter of IkappaBalpha are described in this article. A known PPAR activator (Wy14643) and dimethylsulfoxide (vehicle) were introduced into control (FVB) and delta337T thyroid hormone receptor (TRbeta) transgenic mice. The delta337T TRbeta transgenic mouse has a resistance to the thyroid hormone (RTH) phenotype. Affymetrix 430_2 chip gene expression was examined for four study groups (control, control with Wy14643, delta337T TRbeta and delta337T TRbeta with Wy14643), consisting of seven mice each. IkappaBalpha mRNA expression in the Wy14643 control and in transgenic mice was upregulated significantly in microarray (P < 0.05) and quantitative RT-PCR (P < 0.01) analyses. The increase in mRNA level was also accompanied by an increase in IkappaBalpha protein in cells, as measured by Western blot analysis. Duplex oligo-DNAs containing the putative PPAR/RXR motif (AGGTCA/TCCAGT) from the IkappaBalpha promoter were used in gel shift assays to verify the binding of PPAR and RXR to their response elements. pGL4.0 [Luc] constructs of the IkappaBalpha promoter, with and without the PPAR/RXR motifs, were co-transfected with mouse PPAR alpha, beta and gamma(1) into HepG2 cells and used in luciferase assays to verify gene activation. In conclusion, our study revealed that PPAR regulates the mouse cardiac IkappaBalpha gene in both control and transgenic mouse heart. The implications of this finding are discussed in relation to possible changes in cardiac function.


Subject(s)
I-kappa B Proteins/metabolism , Myocardium/metabolism , Peroxisome Proliferator-Activated Receptors/metabolism , Receptors, Thyroid Hormone/metabolism , Animals , Blotting, Western , Cell Line, Tumor , Electrophoretic Mobility Shift Assay , Gene Expression/drug effects , Humans , I-kappa B Proteins/genetics , Luciferases/genetics , Luciferases/metabolism , Mice , Mice, Knockout , Mice, Transgenic , NF-KappaB Inhibitor alpha , Peroxisome Proliferator-Activated Receptors/genetics , Peroxisome Proliferators/pharmacology , Promoter Regions, Genetic/genetics , Protein Binding , Pyrimidines/administration & dosage , Pyrimidines/pharmacology , Receptors, Thyroid Hormone/genetics , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Retinoid X Receptors/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Transfection
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