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1.
Endocrine ; 2024 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-38448677

RESUMO

OBJECTIVES: There is ample that metabolic dysregulation is involved in Graves' disease (GD) and Graves' ophthalmopathy (GO). Recent studies have identified numerous metabolites associated with GD and GO. However, the causal impact of metabolites on GD and GO remains to be investigated. METHODS: This two-sample Mendelian randomization (MR) analysis investigated the causal relationships between 486 blood metabolites and GD and GO. Sensitivity analysis was also performed to examine heterogeneity and pleiotropy. RESULTS: MR analysis showed that 9 and 13 metabolites were associated with GD and GO, respectively, each meeting the nominal significance criteria (inverse variance weighted, p < 0.05). Additionally, four metabolic pathways were identified for each condition using network-based MetaboAnalyst 5.0. CONCLUSIONS: The metabolites and pathways discovered in this study could serve as circulating metabolic biomarkers for clinical screening and prevention of GD and GO. They can be also used for further studies on the mechanisms and drug targets in GD and GO.

2.
Lupus ; 33(3): 223-231, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38214324

RESUMO

OBJECTIVES: Observational studies indicate a significant correlation between systemic lupus erythematosus (SLE) and endocrine and metabolic disorders, but the causal association between SLE and endocrine and metabolic disorders remains unclear due to the reverse causality and confounding biases commonly presented in conventional observational research. This study endeavors to uncover the causal association between SLE and three common endocrine and metabolic disorders, including Graves' disease (GD), type 2 diabetes mellitus (T2DM), and osteoporosis (OP). METHODS: We used genome-wide association study data for SLE and three endocrine and metabolic disorders in an East Asian population, employing bidirectional two-sample Mendelian randomization (MR) analysis and sensitivity analysis to ascertain the causal association between SLE and endocrine and metabolic disorders. RESULTS: A multiplicative random-effect inverse-variance weighted approach revealed a significant positive correlation between SLE and an elevated risk of GD with an odds ratio (OR) of 1.12 (95% CI: 1.04-1.22, p < .01), and inverse-variance weighted (IVW) analysis also indicated that SLE increased the risk of OP with an OR of 1.035 (95% CI: 1.003-1.068, p < .05). Additionally, GD causally affected SLE in an IVW analysis after Bonferroni correction, with an OR of 1.33 (95% CI: 1.19-1.49, p < .05/3), but the application of multivariable MR analysis resulted in the absence of a causal association of GD on SLE (OR 1.047, 95% CI: 0.952-1.151, p > .05). Lastly, the robustness and validity of the findings were verified through a sensitivity analysis. CONCLUSIONS: We confirmed that SLE has a causal effect on GD as well as OP, but no evidence exists to substantiate a causal link between SLE and T2DM. Our study offers valuable contributions for uncovering the etiology of SLE and endocrine and metabolic disorders and furthering disease risk research while providing potential targets for disease monitoring and therapeutic intervention.


Assuntos
Diabetes Mellitus Tipo 2 , Lúpus Eritematoso Sistêmico , Doenças Metabólicas , Osteoporose , Humanos , Diabetes Mellitus Tipo 2/epidemiologia , Diabetes Mellitus Tipo 2/genética , População do Leste Asiático , Estudo de Associação Genômica Ampla , Lúpus Eritematoso Sistêmico/epidemiologia , Lúpus Eritematoso Sistêmico/genética , Análise da Randomização Mendeliana , Doenças Metabólicas/epidemiologia , Doenças Metabólicas/genética , Polimorfismo de Nucleotídeo Único
3.
Wounds ; 35(1): E69-E73, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-37018743

RESUMO

INTRODUCTION: Delayed and incomplete healing of the skin graft at the donor site of an LD flap is common because of seroma formation. The authors aimed to evaluate whether the application of an NPD could improve the healing process after STSG at LD donor sites. MATERIALS AND METHODS: From July 2019 to September 2021, 32 patients underwent STSG with NPD at the LD donor site, and 27 patients underwent STSG with TBDs. Data were collected and analyzed using the chi-square test, t test, and Spearman correlation test. RESULTS: The overall Spearman correlations of graft loss with seroma, hematoma, and infection were 0.56 (P <.01), 0.64 (P <.01), and 0.70 (P <.01), respectively. Compared with the TBD group, the STSG take rate was significantly higher in the NPD group (90.3% vs 84.5%, P =.046) while the seroma rate (18.8% vs 44.4%, P =.033), graft loss (9.4% vs 29.6%, P =.047), and mean length of stay (10.9 ± 1.8 vs 12.1 ± 2.4, P =.037) were significantly lower. CONCLUSIONS: NPDs for STSG at the LD donor site contribute significantly to improved graft acceptance with reduced seroma formation.


Assuntos
Mamoplastia , Tratamento de Ferimentos com Pressão Negativa , Músculos Superficiais do Dorso , Humanos , Transplante de Pele/efeitos adversos , Tratamento de Ferimentos com Pressão Negativa/efeitos adversos , Seroma/etiologia , Músculos Superficiais do Dorso/transplante , Estudos Retrospectivos , Bandagens/efeitos adversos , Mamoplastia/efeitos adversos
4.
Mol Plant ; 16(5): 930-961, 2023 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-36960533

RESUMO

Nuclear proteins are major constituents and key regulators of nucleome topological organization and manipulators of nuclear events. To decipher the global connectivity of nuclear proteins and the hierarchically organized modules of their interactions, we conducted two rounds of cross-linking mass spectrometry (XL-MS) analysis, one of which followed a quantitative double chemical cross-linking mass spectrometry (in vivoqXL-MS) workflow, and identified 24,140 unique crosslinks in total from the nuclei of soybean seedlings. This in vivo quantitative interactomics enabled the identification of 5340 crosslinks that can be converted into 1297 nuclear protein-protein interactions (PPIs), 1220 (94%) of which were non-confirmative (or novel) nuclear PPIs compared with those in repositories. There were 250 and 26 novel interactors of histones and the nucleolar box C/D small nucleolar ribonucleoprotein complex, respectively. Modulomic analysis of orthologous Arabidopsis PPIs produced 27 and 24 master nuclear PPI modules (NPIMs) that contain the condensate-forming protein(s) and the intrinsically disordered region-containing proteins, respectively. These NPIMs successfully captured previously reported nuclear protein complexes and nuclear bodies in the nucleus. Surprisingly, these NPIMs were hierarchically assorted into four higher-order communities in a nucleomic graph, including genome and nucleolus communities. This combinatorial pipeline of 4C quantitative interactomics and PPI network modularization revealed 17 ethylene-specific module variants that participate in a broad range of nuclear events. The pipeline was able to capture both nuclear protein complexes and nuclear bodies, construct the topological architectures of PPI modules and module variants in the nucleome, and probably map the protein compositions of biomolecular condensates.


Assuntos
Arabidopsis , Núcleo Celular , Arabidopsis/genética , Arabidopsis/metabolismo , Espectrometria de Massas , Proteínas Nucleares/metabolismo
5.
Acta Chir Belg ; 122(3): 215-221, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35499197

RESUMO

BACKGROUND: Pedicle length deficiency in microsurgical procedure is a challenging issue. The aim of this report is to present a case series of a flap-splitting technique for pedicle lengthening of large multiple perforator-based (MPB) free flaps. METHODS: In this retrospective case series, we reviewed the medical records of patients who underwent free flap repair with "split flap" pedicle-lengthening technique between August 2017 and December 2021. Main outcome measures included patient demographics, indications, defect size, flap type, additional vascular pedicle length, and flap survival. RESULTS: Data from 16 patients (age 38-78 years) were reviewed. Indications included breast reconstruction, repair of scalp malignancy or titanium mesh implant exposure, and repair of burn scar contracture. Flaps included ALT flap, LD flap and DIEP flap. The mean added pedicle length was 3.8 cm. Fifteen flaps survived completely and one flap was necrotic distally. No major complications occurred. CONCLUSIONS: The 'split flap' technique could be an effective remedy for unexpected pedicle length deficiencies in large MPB free flaps.


Assuntos
Contratura , Retalhos de Tecido Biológico , Mamoplastia , Retalho Perfurante , Procedimentos de Cirurgia Plástica , Adulto , Idoso , Contratura/etiologia , Retalhos de Tecido Biológico/cirurgia , Humanos , Mamoplastia/métodos , Pessoa de Meia-Idade , Retalho Perfurante/irrigação sanguínea , Procedimentos de Cirurgia Plástica/métodos , Estudos Retrospectivos
6.
Adv Mater ; 34(1): e2104704, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34632634

RESUMO

Biomolecular condensates have been demonstrated as a ubiquitous phenomenon in biological systems and play a crucial role in controlling cellular functions. However, the spatiotemporal construction of artificial biomolecular condensates with functions remains challenging and has been less explored. Herein, a general approach is reported to construct biomolecular condensates (e.g., hydrogel) in the lysosome of living cells for cancer therapy and address multiple drug resistance induced by lysosome sequestration. Aromatic-motif-appended pH-responsive hexapeptide (LTP) derived from natural insulin can be uptaken by cancer cells mainly through caveolae-dependent endocytosis, ensuring the proton-triggered phase transformation (solution to hydrogel) of LTP inside the lysosome specifically. Lysosomal hydrogelation further leads to enlargement of the lysosome in cancer cells and increases the permeability of the lysosome, resulting in cancer cell death. Importantly, lysosomal assemblies can significantly improve the efficiency of current chemotherapy drugs toward multidrug resistance (MDR) cells in vitro and in xenograft tumor models. As an example of functional artificial condensates in lysosomes, this work provides a new strategy for controlling functional condensates formation precisely in the organelles of living cells and addressing MDR in cancer therapy.


Assuntos
Resistencia a Medicamentos Antineoplásicos , Neoplasias , Resistência a Múltiplos Medicamentos , Humanos , Lisossomos/metabolismo , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Oligopeptídeos/metabolismo , Oligopeptídeos/farmacologia
7.
Int J Inj Contr Saf Promot ; 28(4): 403-407, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34404323

RESUMO

Chinese mainland launched the 60-day first-level response to major public health emergencies during the COVID-19 outbreak. This study aimed to determine the incidence and describe the characteristics and predictors of patients who presented to the emergency department (ED) with facial trauma during this period. A retrospective review was conducted on the basis of data on facial trauma presented at the ED of XXX Hospital from 24 January 2020 to 23 March 2020 and the same period in the previous two years. Multivariate linear regression model was employed to explore potential determinants associated with daily number of facial trauma. Significant reduction was observed in the amount of facial trauma during the COVID-19 level I emergency response. The trauma volume evenly distributed over the week. The declined most significantly by age group, 20-29 years, and by time range of visit, 00:00-08:00. Multivariate regression analyses revealed positive relationship between daily minimum temperature and facial trauma volume. The number of facial injuries decreased significantly during the COVID-19 Level 1 emergency response, with the least reduction in total daytime facial trauma and in infant and child facial trauma. And a higher minimum temperature may lead to increased number of facial trauma presentations.


Assuntos
COVID-19 , Traumatismos Faciais , Adulto , Criança , China , Serviço Hospitalar de Emergência , Humanos , Lactente , Estudos Retrospectivos , SARS-CoV-2 , Adulto Jovem
8.
Biomolecules ; 11(8)2021 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-34439883

RESUMO

Isotopically dimethyl labeling was applied in a quantitative post-translational modification (PTM) proteomic study of phosphoproteomic changes in the drought responses of two contrasting soybean cultivars. A total of 9457 phosphopeptides were identified subsequently, corresponding to 4571 phosphoprotein groups and 3889 leading phosphoproteins, which contained nine kinase families consisting of 279 kinases. These phosphoproteins contained a total of 8087 phosphosites, 6106 of which were newly identified and constituted 54% of the current soybean phosphosite repository. These phosphosites were converted into the highly conserved kinase docking sites by bioinformatics analysis, which predicted six kinase families that matched with those newly found nine kinase families. The overly post-translationally modified proteins (OPP) occupies 2.1% of these leading phosphoproteins. Most of these OPPs are photoreceptors, mRNA-, histone-, and phospholipid-binding proteins, as well as protein kinase/phosphatases. The subgroup population distribution of phosphoproteins over the number of phosphosites of phosphoproteins follows the exponential decay law, Y = 4.13e-0.098X - 0.04. Out of 218 significantly regulated unique phosphopeptide groups, 188 phosphoproteins were regulated by the drought-tolerant cultivar under the water loss condition. These significantly regulated phosphoproteins (SRP) are mainly enriched in the biological functions of water transport and deprivation, methionine metabolic processes, photosynthesis/light reaction, and response to cadmium ion, osmotic stress, and ABA response. Seventeen and 15 SRPs are protein kinases/phosphatases and transcription factors, respectively. Bioinformatics analysis again revealed that three members of the calcium dependent protein kinase family (CAMK family), GmSRK2I, GmCIPK25, and GmAKINß1 kinases, constitute a phosphor-relay-mediated signal transduction network, regulating ion channel activities and many nuclear events in this drought-tolerant cultivar, which presumably contributes to the development of the soybean drought tolerance under water deprivation process.


Assuntos
Glycine max/metabolismo , Fosfoproteínas/metabolismo , Proteoma/metabolismo , Proteínas de Soja/metabolismo , Secas , Pressão Osmótica , Fosforilação
9.
Front Cell Dev Biol ; 8: 237, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32351957

RESUMO

Adult stem cells have unique properties in both proliferation and differentiation preference. In this study, we hypothesized that adipose stem cells have a depot-dependent lineage preference. Four rabbits were used to provide donor-matched adipose stem cells from either subcutaneous adipose tissue (ScAT) or infrapatellar fat pad (IPFP). Proliferation and multi-lineage differentiation were evaluated in adipose stem cells from donor-matched ScAT and IPFP. RNA sequencing (RNA-seq) and proteomics were conducted to uncover potential molecular discrepancy in adipose stem cells and their corresponding matrix microenvironments. We found that stem cells from ScAT exhibited significantly higher proliferation and adipogenic capacity compared to those from donor-matched IPFP while stem cells from IPFP displayed significantly higher chondrogenic potential compared to those from donor-matched ScAT. Our findings are strongly endorsed by supportive data from transcriptome and proteomics analyses, indicating a site-dependent lineage preference of adipose stem cells.

10.
Ann Plast Surg ; 85(2): 141-145, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-31913895

RESUMO

BACKGROUND: Size discrepancy in microvascular anastomosis is a common issue in free flap transfer and replantation surgery. A number of different techniques have been described to overcome the problem, but optimal method continues to be defined. METHODS: Since June 2015 to May 2018, clinical courses of 103 microvascular cases performed by one senior surgeon were reviewed. Three end-to-end techniques including mechanical dilation, single-mattress suture, and wedge resection were applied in 364 anastomoses with caliber ratio between 1:1 and 1:1.5, 1:1.5 and 1:2, and 1:2 and 1: 3, respectively. RESULTS: A total of 112 flaps were incorporated in this study. The incidence of anastomotic failure was 3.0% (11/364), and the overall flap failure rate was 3.6% (4/112). The failure cases included 2 replanted scalps, 1 replanted ear, and 1 superficial temporal artery flap for nasal reconstruction. CONCLUSIONS: Our results depicted operational convenience and reliability of the 3 end-to-end anastomotic techniques in addressing mild-to-large vessel discrepancy.


Assuntos
Retalhos de Tecido Biológico , Microcirurgia , Anastomose Cirúrgica , Humanos , Reprodutibilidade dos Testes , Suturas
11.
Nat Chem Biol ; 16(4): 440-449, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31873224

RESUMO

Indole signaling is an important cross-species communication pathway in the mammalian gut. In bacteria, upon induction by tryptophan, the molecular sensor (tnaC) controls indole biosynthesis by precisely coordinating dynamics of the corresponding macromolecular machineries during its transcription and translation. Our understanding of this regulatory program is still limited owing to its rapid dynamic nature. To address this shortcoming, we adopted a massively parallel profiling method to quantify the responses of 1,450 synthetic tnaC variants in the presence of three concentrations of tryptophan in living bacterial cells. The resultant dataset enabled us to comprehensively probe the key intermediate states of macromolecular machineries during the transcription and translation of tnaC. We also used modeling to provide a systems-level understanding of how these critical states collectively shape the output of this regulatory program quantitatively. A similar methodology will likely apply to other poorly understood dynamics-dependent cis-regulatory elements.


Assuntos
Proteínas de Escherichia coli/metabolismo , Indóis/metabolismo , Proteínas de Bactérias/metabolismo , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica/fisiologia , Biossíntese de Proteínas/efeitos dos fármacos , Sinais Direcionadores de Proteínas , Ribossomos/metabolismo , Transdução de Sinais/fisiologia , Transcrição Gênica/efeitos dos fármacos , Triptofano/metabolismo
12.
Aesthetic Plast Surg ; 43(6): 1588-1594, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31555873

RESUMO

BACKGROUND: Treatments including selective neurectomy, muscle resections and botulinum toxin A (BTX) injections have been used to improve the stocky appearance of calves. BTX injection has the advantages of high efficiency and is almost noninvasive. However, criterion standards of injection are still missing. OBJECTIVE: We aimed to establish a method to classify the hypertrophic calf for a personalized treatment and set up an injection protocol based on the findings. METHODS: Three-dimensional CT reconstruction was used to measure the thickness and cross-sectional area of the triceps surae. B-mode ultrasound and palpation were used to evaluate the muscle thickness and determine the dosage. Patients were followed 3 and 6 months after the treatment. RESULTS: A total of 112 legs were classified into three degrees of thickness (< 15 mm, 15-25 mm and > 25 mm). Twenty-seven subjects were treated with an individualized BTX (100-300 U). Maximal circumference decreased 0.33 ± 0.00 cm after 3 month (p < 0.05) and 0.67 ± 0.11 cm after 6 months (p < 0.01). The angulated calf contour was improved. No severe side effects were reported. CONCLUSIONS: Localizing and dosage are the key points when applying BTX. Dosage should be decided by muscle thickness instead of circumference. BTX treatment improves the prominent contour of the calf rather than reducing the volume. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.


Assuntos
Toxinas Botulínicas Tipo A/administração & dosagem , Músculo Esquelético/patologia , Adulto , Feminino , Humanos , Hipertrofia/classificação , Hipertrofia/tratamento farmacológico , Injeções Intralesionais , Perna (Membro) , Adulto Jovem
13.
Science ; 361(6406)2018 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-30093605

RESUMO

Mutations in two genes, PKD1 and PKD2, account for most cases of autosomal dominant polycystic kidney disease, one of the most common monogenetic disorders. Here we report the 3.6-angstrom cryo-electron microscopy structure of truncated human PKD1-PKD2 complex assembled in a 1:3 ratio. PKD1 contains a voltage-gated ion channel (VGIC) fold that interacts with PKD2 to form the domain-swapped, yet noncanonical, transient receptor potential (TRP) channel architecture. The S6 helix in PKD1 is broken in the middle, with the extracellular half, S6a, resembling pore helix 1 in a typical TRP channel. Three positively charged, cavity-facing residues on S6b may block cation permeation. In addition to the VGIC, a five-transmembrane helix domain and a cytosolic PLAT domain were resolved in PKD1. The PKD1-PKD2 complex structure establishes a framework for dissecting the function and disease mechanisms of the PKD proteins.


Assuntos
Complexos Multiproteicos/química , Rim Policístico Autossômico Dominante/metabolismo , Canais de Cátion TRPP/química , Microscopia Crioeletrônica , Cristalografia por Raios X , Humanos , Complexos Multiproteicos/genética , Complexos Multiproteicos/ultraestrutura , Mutação , Rim Policístico Autossômico Dominante/genética , Domínios Proteicos , Dobramento de Proteína , Canais de Cátion TRPP/genética , Canais de Cátion TRPP/metabolismo , Canais de Cátion TRPP/ultraestrutura
14.
J Proteome Res ; 17(9): 3195-3213, 2018 09 07.
Artigo em Inglês | MEDLINE | ID: mdl-30084631

RESUMO

An in planta chemical cross-linking-based quantitative interactomics (IPQCX-MS) workflow has been developed to investigate in vivo protein-protein interactions and alteration in protein structures in a model organism, Arabidopsis thaliana. A chemical cross-linker, azide-tag-modified disuccinimidyl pimelate (AMDSP), was directly applied onto Arabidopsis tissues. Peptides produced from protein fractions of CsCl density gradient centrifugation were dimethyl-labeled, from which the AMDSP cross-linked peptides were fractionated on chromatography, enriched, and analyzed by mass spectrometry. ECL2 and SQUA-D software were used to identify and quantitate these cross-linked peptides, respectively. These computer programs integrate peptide identification with quantitation and statistical evaluation. This workflow eventually identified 354 unique cross-linked peptides, including 61 and 293 inter- and intraprotein cross-linked peptides, respectively, demonstrating that it is able to in vivo identify hundreds of cross-linked peptides at an organismal level by overcoming the difficulties caused by multiple cellular structures and complex secondary metabolites of plants. Coimmunoprecipitation and super-resolution microscopy studies have confirmed the PHB3-PHB6 protein interaction found by IPQCX-MS. The quantitative interactomics also found hormone-induced structural changes of SBPase and other proteins. This mass-spectrometry-based interactomics will be useful in the study of in vivo protein-protein interaction networks in agricultural crops and plant-microbe interactions.


Assuntos
Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Mapeamento de Interação de Proteínas/métodos , Proteoma/metabolismo , Proteínas Repressoras/metabolismo , Sequência de Aminoácidos , Arabidopsis/genética , Proteínas de Arabidopsis , Cromatografia Líquida , Reagentes de Ligações Cruzadas/química , Modelos Moleculares , Peptídeos/análise , Peptídeos/química , Proibitinas , Ligação Proteica , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Estrutura Secundária de Proteína , Proteólise , Proteoma/química , Proteoma/genética , Proteínas Repressoras/química , Proteínas Repressoras/genética , Coloração e Rotulagem/métodos , Succinimidas/química , Espectrometria de Massas em Tandem
15.
Nat Commun ; 9(1): 1192, 2018 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-29567962

RESUMO

PKD2L1, also termed TRPP3 from the TRPP subfamily (polycystic TRP channels), is involved in the sour sensation and other pH-dependent processes. PKD2L1 is believed to be a nonselective cation channel that can be regulated by voltage, protons, and calcium. Despite its considerable importance, the molecular mechanisms underlying PKD2L1 regulations are largely unknown. Here, we determine the PKD2L1 atomic structure at 3.38 Å resolution by cryo-electron microscopy, whereby side chains of nearly all residues are assigned. Unlike its ortholog PKD2, the pore helix (PH) and transmembrane segment 6 (S6) of PKD2L1, which are involved in upper and lower-gate opening, adopt an open conformation. Structural comparisons of PKD2L1 with a PKD2-based homologous model indicate that the pore domain dilation is coupled to conformational changes of voltage-sensing domains (VSDs) via a series of π-π interactions, suggesting a potential PKD2L1 gating mechanism.


Assuntos
Canais de Cálcio/química , Receptores de Superfície Celular/química , Animais , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Canais de Cálcio/ultraestrutura , Microscopia Crioeletrônica , Humanos , Camundongos , Conformação Proteica em alfa-Hélice , Domínios Proteicos , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Receptores de Superfície Celular/ultraestrutura , Canais de Cátion TRPP/química , Canais de Cátion TRPP/genética , Canais de Cátion TRPP/metabolismo
16.
Mol Cell Proteomics ; 17(5): 1010-1027, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29440448

RESUMO

Protein acetylation, one of many types of post-translational modifications (PTMs), is involved in a variety of biological and cellular processes. In the present study, we applied both CsCl density gradient (CDG) centrifugation-based protein fractionation and a dimethyl-labeling-based 4C quantitative PTM proteomics workflow in the study of dynamic acetylproteomic changes in Arabidopsis. This workflow integrates the dimethyl chemical labeling with chromatography-based acetylpeptide separation and enrichment followed by mass spectrometry (MS) analysis, the extracted ion chromatogram (XIC) quantitation-based computational analysis of mass spectrometry data to measure dynamic changes of acetylpeptide level using an in-house software program, named Stable isotope-based Quantitation-Dimethyl labeling (SQUA-D), and finally the confirmation of ethylene hormone-regulated acetylation using immunoblot analysis. Eventually, using this proteomic approach, 7456 unambiguous acetylation sites were found from 2638 different acetylproteins, and 5250 acetylation sites, including 5233 sites on lysine side chain and 17 sites on protein N termini, were identified repetitively. Out of these repetitively discovered acetylation sites, 4228 sites on lysine side chain (i.e. 80.5%) are novel. These acetylproteins are exemplified by the histone superfamily, ribosomal and heat shock proteins, and proteins related to stress/stimulus responses and energy metabolism. The novel acetylproteins enriched by the CDG centrifugation fractionation contain many cellular trafficking proteins, membrane-bound receptors, and receptor-like kinases, which are mostly involved in brassinosteroid, light, gravity, and development signaling. In addition, we identified 12 highly conserved acetylation site motifs within histones, P-glycoproteins, actin depolymerizing factors, ATPases, transcription factors, and receptor-like kinases. Using SQUA-D software, we have quantified 33 ethylene hormone-enhanced and 31 hormone-suppressed acetylpeptide groups or called unique PTM peptide arrays (UPAs) that share the identical unique PTM site pattern (UPSP). This CDG centrifugation protein fractionation in combination with dimethyl labeling-based quantitative PTM proteomics, and SQUA-D may be applied in the quantitation of any PTM proteins in any model eukaryotes and agricultural crops as well as tissue samples of animals and human beings.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteômica/métodos , Coloração e Rotulagem , Acetilação , Sequência de Aminoácidos , Cromatografia Líquida , Biologia Computacional , Etilenos/farmacologia , Histonas/metabolismo , Metilação , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem
17.
Cell Res ; 27(8): 1020-1033, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28621327

RESUMO

The Escherichia coli uracil:proton symporter UraA is a prototypical member of the nucleobase/ascorbate transporter (NAT) or nucleobase/cation symporter 2 (NCS2) family, which corresponds to the human solute carrier family SLC23. UraA consists of 14 transmembrane segments (TMs) that are organized into two distinct domains, the core domain and the gate domain, a structural fold that is also shared by the SLC4 and SLC26 transporters. Here we present the crystal structure of UraA bound to uracil in an occluded state at 2.5 Å resolution. Structural comparison with the previously reported inward-open UraA reveals pronounced relative motions between the core domain and the gate domain as well as intra-domain rearrangement of the gate domain. The occluded UraA forms a dimer in the structure wherein the gate domains are sandwiched by two core domains. In vitro and in vivo biochemical characterizations show that UraA is at equilibrium between dimer and monomer in all tested detergent micelles, while dimer formation is necessary for the transport activity. Structural comparison between the dimeric UraA and the recently reported inward-facing dimeric UapA provides important insight into the transport mechanism of SLC23 transporters.


Assuntos
Escherichia coli O157/química , Proteínas de Escherichia coli/química , Proteínas de Membrana Transportadoras/química , Multimerização Proteica , Proteínas de Transporte de Ânions/química , Proteínas de Transporte de Ânions/genética , Escherichia coli O157/genética , Proteínas de Escherichia coli/genética , Humanos , Proteínas de Membrana Transportadoras/genética , Domínios Proteicos , Estrutura Quaternária de Proteína , Homologia Estrutural de Proteína
18.
F1000Res ; 6: 291, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28413622

RESUMO

For decades, researchers have been fascinated by the strategy of using cell therapy for bone defects; some progress in the field has been made. Owing to its ample supply and easy access, skin, the largest organ in the body, has gained attention as a potential source of stem cells. Despite extensive applications in skin and nerve regeneration, an increasing number of reports indicate its potential use in bone tissue engineering and regeneration. Unfortunately, few review articles are available to outline current research efforts in skin-based osteogenesis. This review first summarizes the latest findings on stem cells or progenitors in skin and their niches and then discusses the strategies of skin cell-based osteogenesis. We hope this article elucidates this topic and generates new ideas for future studies.

19.
PLoS One ; 12(2): e0171651, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28187214

RESUMO

BACKGROUND: We developed a novel pedicled DIEP flap model in rat to explore the possible remedy for the distal necrosis of the flap. METHODS: A deep inferior epigastric perforator (DIEP) flap, based on the second right cranial perforator (P2) as the main pedicle, was elevated in 48 Sprague-Dawley rats. The rats were randomized into 4 groups: group I, the left P2 remaining intact as supercharging; group II, the left P2 artery alone kept as supercharging; group III, the left P2 vein alone kept as supercharging; group IV, no supercharging. Transcutaneous oxygen pressure (TcPO2) and transcutaneous carbon dioxide pressure (TcPCO2) were measured immediately after flap elevation, protein level of Hif-1a was measured 48 hours later, and flap survival was assessed 7 days postoperatively. RESULTS: Blockade of artery led to significantly lower TcPO2, higher TcPCO2, and higher expression level of Hif-1a in the distal side of the flap in group III and group IV, than those of group I and group II. At 7 days post surgery, significantly lower flap survival rates were observed in group III (81.9 ± 5.7%) and group IV (78.4 ± 6.5%), compared to observed in group I (97.2 ± 3.0%) and group II (94.2 ± 6.2%). CONCLUSIONS: It might be arterial insufficiency, not venous congestion, which mainly caused the distal necrosis of the DIEP flap in rat. Arterial instead of venous supercharging might be a more effective procedure that improves circulation to zone IV of the flap.


Assuntos
Infarto/prevenção & controle , Retalho Perfurante/irrigação sanguínea , Procedimentos de Cirurgia Plástica/métodos , Complicações Pós-Operatórias/prevenção & controle , Animais , Drenagem/métodos , Infarto/etiologia , Masculino , Retalho Perfurante/efeitos adversos , Retalho Perfurante/patologia , Complicações Pós-Operatórias/etiologia , Ratos , Ratos Sprague-Dawley , Procedimentos de Cirurgia Plástica/efeitos adversos , Fluxo Sanguíneo Regional
20.
Stem Cells Transl Med ; 6(1): 306-315, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-28170187

RESUMO

Finding appropriate seed cells for bone tissue engineering remains a significant challenge. Considering that skin is the largest organ, we hypothesized that human bone morphogenetic protein receptor type IB (BmprIB)+ dermal cells could have enhanced osteogenic capacity in the healing of critical-sized calvarial defects in an immunodeficient mouse model. In this study, immunohistochemical staining revealed that BmprIB was expressed throughout reticular dermal cells; the positive expression rate of BmprIB was 3.5% ± 0.4% in freshly separated dermal cells, by flow cytometry. Furthermore, in vitro osteogenic capacity of BmprIB+ cells was confirmed by osteogenic-related staining and marker gene expression compared with unsorted dermal cells. In vivo osteogenic capacity was demonstrated by implantation of human BmprIB+ cell/coral constructs in the treatment of 4-mm diameter calvarial defects in an immunodeficient mouse model compared with implantation of unsorted cell/coral constructs and coral scaffold alone. These results indicate that the selective cell population BmprIB from human dermis is a promising osteogenic progenitor cell that can be a large-quantity and high-quality cell source for bone tissue engineering and regeneration. Stem Cells Translational Medicine 2017;6:306-315.


Assuntos
Regeneração Óssea/fisiologia , Derme/citologia , Animais , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/metabolismo , Criança , Pré-Escolar , Humanos , Lactente , Masculino , Camundongos , Osteogênese , Alicerces Teciduais/química , Tomografia Computadorizada por Raios X
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