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1.
Materials (Basel) ; 16(10)2023 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-37241330

RESUMO

Textile-reinforced concrete (TRC) is highly anticipated as an alternative to reinforced concrete due to its ability to enable lightweight design, free formability, and improved ductility. In this study, TRC panel test specimens were fabricated and four-point loading flexural tests were performed to examine the flexural characteristics of TRC panels reinforced with carbon fabric, and to investigate the effect of the fabric reinforcement ratio, anchorage length, and surface treatment of fabric. Furthermore, the flexural behavior of the test specimens was numerically analyzed using the general section analysis concept of reinforced concrete and compared with the experimental results. Due to bond failure between the carbon fabric and the concrete matrix, the TRC panel showed a large decrease in flexural performance in terms of flexural stiffness, flexural strength, cracking behavior, and deflection. This low performance was improved by increasing the fabric reinforcement ratio, anchoring length, and sand-epoxy surface treatment of the anchorage. Comparing the numerical calculation results with the experimental results, the deflection of the experimental results was approximately 50% larger than the numerical calculation results. This is because the perfect bond between the carbon fabric and the concrete matrix failed, and slip occurred.

2.
J Environ Manage ; 340: 117895, 2023 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-37121008

RESUMO

In efforts to minimize environmental pollution and carbon-based gas emissions, photocatalytic hydrogen production and sensing applications at ambient temperature are important. This research reports on the development of new 0D/1D materials based on TiO2 nanoparticles grown onto CdS hetersturctured nanorods via two-stage facile synthesis. The titanate nanoparticles when loaded onto CdS surfaces at an optimized concentration (20 mM), exhibited superior photocatalytic hydrogen production (21.4 mmol/h/gcat). The optimized nanohybrid was recycled for 6 cycles up to 4 h, indicating its excellent stabity for a prolonged period. Also, the photoelectrochemical water oxidation in alkaline medium was investigated to offer the optimized CRT-2 composite with 1.91 mA/cm2@0.8 V vs. RHE (0 V vs. Ag/AgCl) that was used for effective room-temperature NO2 gas detection exhibiting a higher response (69.16%) to NO2 (100 ppm) at room temperature at the lowest detection limit of ∼118 ppb than the pristine counterparts. Further, NO2 gas sensing performance of CRT-2 sensor was increased using UV light (365 nm) activation energy. Under the UV light, the sensor exhibited a remarkable gas sensing response quick response/recovery times (68/74), excellent long-term cycling stability, and significant selectivity to NO2 gas. Due to high porosity and surface area values of CdS (5.3), TiO2 (35.5), and CRT-2 (71.5 m2/g), excellent photocatalytic H2 production and gas sensing of CRT-2 is ascribed to morphology, synergistic effect, improved charge generation, and separation. Overall, 1D/0D CdS@TiO2 is proved to be an efficient material for hydrogen production and gas detection.


Assuntos
Ciclismo , Dióxido de Nitrogênio , Carbono , Hidrogênio
4.
Materials (Basel) ; 16(1)2022 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-36614373

RESUMO

This study developed a 3D concrete printing (3DCP) system that can print not only in air but also underwater. This underwater 3DCP system is equipped with many distinct technologies, such as a technology to supply the printing material to the nozzle tip at a constant rate by detecting its amount in the printer hopper. Using the developed 3DCP system, the effect of nozzle details on underwater print quality and hardened properties was investigated. The straight-line printing performance underwater was evaluated using five nozzles: a nozzle without a trowel (Nozzel#1), a nozzle with fixed trowels attached to both sides (Nozzle#2), a nozzle with trowels attached to the back and both sides to constrain five sides (Nozzle#3), a nozzle with a three-sided trowel inclined by 30° (Nozzle#4), and a nozzle with a roof added to Nozzle#4 opening (Nozzle#5). Nozzle#4 yielded the best print quality and hardened properties. In addition, an underwater curved shape printing test was performed using Nozzle#4, the problems that occurred in this test were analyzed and solutions were suggested.

5.
Materials (Basel) ; 14(24)2021 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-34947358

RESUMO

In this study, we experimentally analyzed the deformation shape of stacked layers developed using three-dimensional (3D) printing technology. The nozzle traveling speed was changed to 80, 90, 100, and 110 mm/s when printing the layers to analyze its effect on layer deformation. Furthermore, the cross-sectional area and the number of layers were analyzed by printing five layers with overall dimensions of 1000 (w) × 2200 (l) × 50 (h) mm (each layer was 10 mm high) using Vernier calipers. Moreover, we analyzed the interface and cross-sectional area of layers that are difficult to confirm visually using X-ray computed tomography (X-ray CT) analysis. As a result of measuring the deformation at the center of the layer, it was confirmed that the deformation was greater for lower nozzle traveling speeds. Consequently, the X-ray CT analysis verified that the layer had the same cross-sectional area irrespective of the layer printing order at the same nozzle travel speed, even if the layer was deformed.

6.
Protein Pept Lett ; 28(12): 1408-1424, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34749602

RESUMO

BACKGROUND: We investigated melanogenesis- and anti-apoptosis-related melanoma factors in melanoma cells (TXM1, TXM18, A375P, and A375SM). OBJECTIVE: To find melanoma associated hub factor, high-throughput screening-based techniques integrating with bioinformatics were investigated. METHODS: Array CGH analysis was conducted with a commercial system. Total genomic DNAs prepared individually from each cell line with control DNA were properly labeled with Cy3-dCTP and Cy5-dCTP and hybridizations and subsequently performed data treatment by the log2 green (G; test) to red (R; reference) fluorescence ratios (G/R). Gain or loss of copy number was judged by spots with log2-transformed ratios. PPI mapping analysis of detected candidate genes based on the array CGH results was conducted using the human interactome in the STRING database. Energy minimization and a short Molecular Dynamics (MD) simulation using the implicit solvation model in CHARMM were performed to analyze the interacting residues between YWHAZ and YWHAB. RESULTS: Three genes (BMP-4, BFGF, LEF-1) known to be involved in melanogenesis were found to lose chromosomal copy numbers, and Chr. 6q23.3 was lost in all tested cell lines. Ten hub genes (CTNNB1, PEX13, PEX14, PEX5, IFNG, EXOSC3, EXOSC1, EXOSC8, UBC, and PEX10) were predicted to be functional interaction factors in the network of the 6q23.3 locus. The apoptosis-associated genes E2F1, p50, BCL2L1, and BIRC7 gained, and FGF2 lost chromosomal copy numbers in the tested melanoma cell lines. YWHAB, which gained chromosomal copy numbers, was predicted to be the most important hub protein in melanoma cells. Molecular dynamics simulations for binding YWHAB and YWHAZ were conducted, and the complex was predicted to be energetically and structurally stable through its 3 hydrogen-bond patterns. The number of interacting residues is 27. CONCLUSION: Our study compares genome-wide screening interactomics predictions for melanoma factors and offers new information for understanding melanogenesis- and anti-apoptosis-associated mechanisms in melanoma. Especially, YWHAB was newly detected as a core factor in melanoma cells.


Assuntos
Proteínas Reguladoras de Apoptose , Regulação Neoplásica da Expressão Gênica , Melanoma , Proteínas de Neoplasias , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas Reguladoras de Apoptose/biossíntese , Proteínas Reguladoras de Apoptose/genética , Linhagem Celular Tumoral , Humanos , Melanoma/genética , Melanoma/metabolismo , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética
7.
Materials (Basel) ; 14(19)2021 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-34640284

RESUMO

Research and technological advancements in 3D concrete printing (3DCP) have led to the idea of applying it to offshore construction. The effect of gravity is reduced underwater, which can have a positive effect on 3DCP. For basic verification of this idea, this study printed and additively manufactured specimens with the same mortar mixture in air and underwater and evaluated properties in the fresh state and the hardened state. The mechanical properties were evaluated using the specimens produced by direct casting to the mold and specimens produced by extracting from the additive part through coring and cutting. The results of the experiment show that underwater 3D printing required a greater amount of printing output than in-air 3D printing for a good print quality, and buildability was improved underwater compared to that in air. In the case of the specimen layered underwater, the density and compressive strength decreased compared to the specimen layered in air. Because there are almost no effects of moisture evaporation and bleeding in water, the interlayer bond strength of the specimen printed underwater was somewhat larger than that printed in air, while there was no effect of the deposition time interval underwater.

8.
Ann Dermatol ; 33(4): 333-338, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34341634

RESUMO

BACKGROUND: Prurigo nodularis (PN) is a highly pruritic disease that significantly impairs patient quality of life. Although the mechanism that causes pruritus is not clear, one hypothesis argues that neural hyperplasia, mast cell, and Merkel cell neurite complexes may be associated with PN pathogenesis. OBJECTIVE: The objective of this study was to analyze whether special staining outcomes differed depending on the presence of atopic dermatitis (AD) and treatment response. METHODS: A total of 209 patients diagnosed with PN was analyzed retrospectively. Patients were divided into two groups according to presence or past history of AD and by treatment response. Histopathologic features were obtained using the following stains: Giemsa, S-100, neuron-specific enolase, cytokeratin (CK)-20, CAM5.2, and CK8/CK18. RESULTS: A total of 126 patients (60.29%) had AD, and 68 (32.54%) showed clinical improvement. There were no statistically significant differences in the staining results between the PN groups with AD (PN c AD) and without AD (PN s AD). Additionally, there were no statistically significant differences in staining results between the improved and non-improved groups. CONCLUSION: Implementing the special stains helped to identify PN pathogenesis. Because there were no statistically significant differences in the special stain results between the improved and non-improved groups, we conclude that mast cell proliferation, neural hyperplasia, and Merkel cell hyperplasia may not have a significant effect on treatment response.

9.
Commun Biol ; 4(1): 692, 2021 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-34099859

RESUMO

Research on human nail tissue has been limited by the restricted access to fresh specimen. Here, we studied transcriptome profiles of human nail units using polydactyly specimens. Single-cell RNAseq with 11,541 cells from 4 extra digits revealed nail-specific mesenchymal and epithelial cell populations, characterized by RSPO4 (major gene in congenital anonychia) and SPINK6, respectively. In situ RNA hybridization demonstrated the localization of RSPO4, MSX1 and WIF1 in onychofibroblasts suggesting the activation of WNT signaling. BMP-5 was also expressed in onychofibroblasts implicating the contribution of BMP signaling. SPINK6 expression distinguished the nail-specific keratinocytes from epidermal keratinocytes. RSPO4+ onychofibroblasts were distributed at close proximity with LGR6+ nail matrix, leading to WNT/ß-catenin activation. In addition, we demonstrated RSPO4 was overexpressed in the fibroblasts of onychomatricoma and LGR6 was highly expressed at the basal layer of the overlying epithelial component, suggesting that onychofibroblasts may play an important role in the pathogenesis of onychomatricoma.


Assuntos
Unhas/citologia , Inibidores de Serinopeptidase do Tipo Kazal/genética , Trombospondinas/genética , Células Cultivadas , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibroblastos/patologia , Humanos , Queratinócitos/citologia , Queratinócitos/metabolismo , Queratinócitos/patologia , Unhas/metabolismo , Unhas/patologia , Análise de Sequência de RNA , Análise de Célula Única , Transcriptoma
10.
Int J Biol Macromol ; 170: 150-163, 2021 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-33359255

RESUMO

We investigated the tyrosinase-associated melanogenesis in melanoma cells by using OMICS techniques. We characterized the chromosome copy numbers, including Chr 11q21 where the tyrosinase gene is located, from several melanoma cell lines (TXM13, G361, and SK-MEL-28) by using array CGH. We revealed that 11q21 is stable in TXM13 cells, which is directly related to a spontaneous high melanin pigment production. Meanwhile, significant loss of copy number of 11q21 was found in G361 and SK-MEL-28. We further profiled the proteome of TXM13 cells by LC-ESI-MSMS and detected more than 900 proteins, then predicted 11 hub proteins (YWHAZ; HSP90AA1; HSPA5; HSPA1L; HSPA9; HSP90B1; HSPA1A; HSPA8; FKSG30; ACTB; DKFZp686DQ972) by using an interactomic algorithm. YWHAZ (25% interaction in the network) is thought to be a most important protein as a linking factor between tyrosinase-triggered melanogenesis and melanoma growth. Bioinformatic tools were further applied for revealing various physiologic mechanisms and functional classification. The results revealed clues for the spontaneous pigmentation capability of TXM13 cells, contrary to G361 and SK-MEL-28 cells, which commonly have depigmentation properties during subculture. Our study comparatively conducted the genome-wide screening and proteomic profiling integrated interactomics prediction for TXM13 cells and suggests new insights for studying both melanogenesis and melanoma.


Assuntos
Hibridização Genômica Comparativa , Biologia Computacional/métodos , Melaninas/biossíntese , Melanoma/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Proteínas de Neoplasias/metabolismo , Proteômica/métodos , Linhagem Celular Tumoral , Cromatografia Líquida , Cromossomos Humanos Par 11/genética , Células Clonais , Chaperona BiP do Retículo Endoplasmático , Dosagem de Genes , Ontologia Genética , Humanos , Melanoma/genética , Monofenol Mono-Oxigenase/genética , Proteínas de Neoplasias/genética , Pigmentação , Mapeamento de Interação de Proteínas , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem
11.
J Biomol Struct Dyn ; 39(6): 2133-2151, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32189581

RESUMO

The function of acetaldehyde dehydrogenase 1 (ALDH1) has been gradually elucidated in several diseases, especially in various cancers. However, the role of ALDH1 in skin-related diseases has been mostly unknown. Previously, we found that ALDH1 is involved in the pathogenesis of atopic dermatitis (AD). In this study, we used high-throughput screening (HTS) approaches to identify critical factors associated with ALDH1 in human keratinocytes to reveal its functions in skin. We overexpressed ALDH1 in human HaCaT keratinocytes and then conducted serial HTS studies, a DNA microarray and antibody array integrated with bioinformatics algorithms. Together, those tests identified several novel genes associated with the function of ALDH1 in keratinocytes, as well as AD, including CTSG and CCL11. In particular, GNB3, GHSR, TAS2R9, FFAR1, TAS2R16, CCL21, GPR32, NPFFR1, GPR15, FBXW12, CCL19, EDNRA, FFAR3, and RXFP3 proteins were consistently detected as hub proteins in the PPI maps. By integrating the datasets obtained from these HTS studies and using the strengths of each method, we obtained new insights into the functional role of ALDH1 in skin keratinocytes. The approach used here could contribute to the clinical understanding of ALDH1-associated applications for the treatment of AD.Communicated by Ramaswamy H. Sarma.


Assuntos
Família Aldeído Desidrogenase 1 , Biologia Computacional , Dermatite Atópica , Retinal Desidrogenase , Humanos , Queratinócitos , Análise em Microsséries
12.
Protein Pept Lett ; 28(1): 84-100, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32586242

RESUMO

BACKGROUND: Previously, we detected that chloride intracellular channel 1 (CLIC1) was involved in the pathogenesis of atopic dermatitis (AD). OBJECTIVE: In this study, we aimed to use high-throughput screening (HTS) approaches to identify critical factors associated with the function of CLIC1 in knock-down cells. METHODS: We down-regulated CLIC1 in human A549 cells via siRNA and then conducted serial HTS studies, including proteomics integrated with a microarray and the implementation of bioinformatics algorithms. RESULTS: Together, these approaches identified several important proteins and genes associated with the function of CLIC1. These proteins and genes included tumor rejection antigen (gp96) 1, nucleophosmin, annexin I, keratin 1 and 10, FLNA protein, enolase 1, and metalloprotease 1, which were found using two-dimensional electrophoresis (2-DE) proteomics. Separately, NTNG1, SEMA5A, CLEC3A, GRPR, GNGT2, GRM5, GRM7, DNMT3B, CXCR5, CCL11, CD86, IL2, MNDA, TLR5, IL23R, DPP6, DLGAP1, CAT, GSTA1, GSTA2, GSTA5, CYP2E1, ADH1A, ESR1, ARRDC3, A1F1, CCL5, CASP8, DNTT, SQSTM1, PCYT1A, and SLCO4C1 were found using a DNA microarray integrated with PPI mapping. CONCLUSION: CCL11 is thought to be a particularly critical gene among the candidate genes detected in this study. By integrating the datasets and utilizing the strengths of HTS, we obtained new insights into the functional role of CLIC1, including the use of CLIC1-associated applications in the treatment of human diseases such as AD.


Assuntos
Canais de Cloreto/metabolismo , Dermatite Atópica/metabolismo , Regulação da Expressão Gênica , Análise Serial de Proteínas , Proteômica , Células A549 , Canais de Cloreto/genética , Dermatite Atópica/genética , Dermatite Atópica/patologia , Técnicas de Silenciamento de Genes , Humanos
13.
Protein Pept Lett ; 28(6): 651-664, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33183186

RESUMO

BACKGROUND: Fibrinolytic protease from Euphausia superba (EFP) was isolated. OBJECTIVE: Biochemical distinctions, regulation of the catalytic function, and the key residues of EFP were investigated. METHODS: The serial inhibition kinetic evaluations coupled with measurements of fluorescence spectra in the presence of 4-(2-aminoethyl) benzene sulfonyl fluoride hydrochloride (AEBSF) was conducted. The computational molecular dynamics (MD) simulations were also applied for a comparative study. RESULTS: The enzyme behaved as a monomeric protein with a molecular mass of about 28.6 kD with Km BApNA = 0.629 ± 0.02 mM and kcat/Km BApNA = 7.08 s-1/mM. The real-time interval measurements revealed that the inactivation was a first-order reaction, with the kinetic processes shifting from a monophase to a biphase. Measurements of fluorescence spectra showed that serine residue modification by AEBSF directly caused conspicuous changes of the tertiary structures and exposed hydrophobic surfaces. Some osmolytes were applied to find protective roles. These results confirmed that the active region of EFP is more flexible than the overall enzyme molecule and serine, as the key residue, is associated with the regional unfolding of EFP in addition to its catalytic role. The MD simulations were supportive to the kinetics data. CONCLUSION: Our study indicated that EFP has an essential serine residue for its catalyst function and associated folding behaviors. Also, the functional role of osmolytes such as proline and glycine that may play a role in defense mechanisms from environmental adaptation in a krill's body was suggested.


Assuntos
Proteínas de Artrópodes , Euphausiacea/enzimologia , Serina Proteases , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/isolamento & purificação , Proteínas de Artrópodes/metabolismo , Fibrinólise , Cinética , Simulação de Dinâmica Molecular , Dobramento de Proteína , Serina Proteases/química , Serina Proteases/isolamento & purificação , Serina Proteases/metabolismo
14.
J Biomol Struct Dyn ; 38(10): 2884-2897, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31345118

RESUMO

In this study, we aimed to identify critical factors associated with superoxide dismutase 2 (SOD2) in human keratinocytes through gene and protein expression profiling approaches. After recombinant SOD2 was exogenously added to culture media, we conducted serial OMICS studies, which included RNA sequencing analysis, integrated antibody-chip arrays, and the implementation of bioinformatics algorithms, in order to reveal genes and proteins that are possibly associated with SOD2 in keratinocytes. These approaches identified several novel genes and proteins in keratinocytes that are associated with exogenous SOD2. These novel genes included DCT, which was up-regulated, and CD38, GPR151, HCK, KIT, and AFP, which were down-regulated. Among them, CD38 and KIT were also predicted as hub proteins in PPI mappings. By integrating the datasets obtained from these complementary high-throughput OMICS studies and utilizing the strengths of each method, we obtained new insights into the functional role of externally added SOD2 in skin cells and into several critical genes that are thought to play important roles in SOD2-associated skin function. The approach used here could help contribute to our clinical understanding of SOD2-associated applications and may be broadly applicable to a wider range of diseases. AbbreviationsSOD2superoxide dismutase 2DAVIDthe database for annotation, visualization and integrated discoveryKEGGKyoto Encyclopedia of Genes and GenomesPPIprotein-protein interactionsHTSHigh-throughput screeningCommunicated by Ramaswamy H. Sarma.


Assuntos
Biologia Computacional , Superóxido Dismutase , Humanos , Queratinócitos , Análise de Sequência de RNA , Superóxido Dismutase/genética
15.
J Biomol Struct Dyn ; 38(12): 3496-3503, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31448679

RESUMO

The inhibition of α-glucosidase is used as a key clinical approach to treat type 2 diabetes mellitus and thus, we assessed the inhibitory effect of α-ketoglutaric acid (AKG) on α-glucosidase with both an enzyme kinetic assay and computational simulations. AKG bound to the active site and interacted with several key residues, including ASP68, PHE157, PHE177, PHE311, ARG312, TYR313, ASN412, ILE434 and ARG439, as detected by protein-ligand docking and molecular dynamics simulations. Subsequently, we confirmed the action of AKG on α-glucosidase as mixed-type inhibition with reversible and rapid binding. The relevant kinetic parameter IC50 was measured (IC50 = 1.738 ± 0.041 mM), and the dissociation constant was determined (Ki Slope = 0.46 ± 0.04 mM). Regarding the relationship between structure and activity, a high AKG concentration induced the slight modulation of the shape of the active site, as monitored by hydrophobic exposure. This tertiary conformational change was linked to AKG inhibition and mostly involved regional changes in the active site. Our study provides insight into the functional role of AKG due to its structural property of a hydroxyphenyl ring that interacts with the active site. We suggest that similar hydroxyphenyl ring-containing compounds targeting key residues in the active site might be potential α-glucosidase inhibitors. AbbreviationsAKGalpha-ketoglutaric acidpNPG4-nitrophenyl-α-d-glucopyranosideANS1-anilinonaphthalene-8-sulfonateMDmolecular dynamics.Communicated by Ramaswamy H. Sarma.


Assuntos
Inibidores de Glicosídeo Hidrolases/farmacologia , Ácidos Cetoglutáricos/farmacologia , alfa-Glucosidases , Diabetes Mellitus Tipo 2 , Humanos , Cinética , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , alfa-Glucosidases/metabolismo
16.
J Biomol Struct Dyn ; 38(9): 2633-2649, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31244373

RESUMO

Previously, we detected that 14-3-3 protein epsilon (YWHAE) was involved in the pathogenesis of atopic dermatitis (AD) and tyrosinase-mediated pigmentation. In this study, we aimed to identify critical factors associated with YWHAE in human keratinocytes using high-throughput screening (HTS) approaches to reveal its functions in skin. We overexpressed YWHAE in human HaCaT keratinocytes and then conducted serial HTS studies, including RNA sequencing integrated with antibody arrays and the implementation of bioinformatics algorithms. Cumulatively, these approaches identified several novel genes in keratinocytes associated with the function of YWHAE including KRT9, KRT1, KRT6C, BST2, CIB2, APH1B, ACTC1, IFI27, TUBA1A, CAPN6, UTY, MX2, and MAPK15, based on RNA sequencing data, and MAPK1, MMP2, TYK2, NOS3, and CASP3, based on antibody array data. In particular, CD37 is a unique gene that was detected and validated in all the methods applied in this study. By integrating the datasets obtained from these HTS studies and utilizing the strengths of each method, we obtained new insights into the functional role of YWHAE in skin keratinocytes. The approach used here could contribute to the clinical understanding of YWHAE-associated applications in the treatment of AD disease. AbbreviationsDAVIDthe database for annotation, visualization and integrated discoveryHTSHigh-throughput screeningKEGGKyoto Encyclopedia of Genes and GenomesPPIprotein-protein interactionsCommunicated by Ramaswamy H. Sarma.


Assuntos
Proteínas 14-3-3/metabolismo , Dermatite Atópica , Queratinócitos , Proteínas 14-3-3/genética , Biologia Computacional , Dermatite Atópica/genética , MAP Quinases Reguladas por Sinal Extracelular , Células HaCaT , Humanos , Análise de Sequência de RNA
17.
Int J Biol Macromol ; 133: 391-411, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-30974145

RESUMO

Previously, we have identified the C3dg protein as an important player in the pathogenesis of atopic dermatitis (AD). In this study, we aimed to identify critical factors associated with C3dg in human keratinocytes based on high-throughput screening (HTS) approaches. We overexpressed C3dg in HaCaT human keratinocytes and conducted serial HTS studies, including RNA sequencing analysis integrated with antibody-chip arrays and implementation of bioinformatics algorithms (PPI mappings). Cumulatively, these approaches identified several novel C3dg-associated genes and proteins that are thought to be significantly involved in skin diseases including AD. These novel genes and proteins included LPA, PROZ, BLK, CLDN11, and FGF22, which are believed to play important roles in C3dg-associated skin functions in keratinocytes, as well as genes related to the two important pathways of systemic lupus erythematosus and Staphylococcus aureus infection. In particular, FGF22 is a unique gene that was detected and validated in all methods applied in this study. By integrating the datasets obtained from these HTS studies and utilizing the strengths of each method, we obtained new insights into the functional role of C3dg in keratinocytes. The approach used here contributes to clinical understanding of C3dg-associated applications and may also be applicable to treatment of AD.


Assuntos
Anticorpos/metabolismo , Complemento C3b/genética , Complemento C3b/metabolismo , Biologia Computacional , Queratinócitos/metabolismo , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Análise Serial de Proteínas , Análise de Sequência de RNA , Algoritmos , Células Hep G2 , Humanos
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