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1.
Int J Biol Macromol ; 2020 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-32437800

RESUMO

CD117/c-kit, a tyrosine kinase receptor, plays a critical role in hematopoiesis, pigmentation, and fertility. The overexpression and activation of c-kit are thought to promote tumor growth and have been reported in various cancers, including leukemia, glioblastoma and mastocytosis. To disrupt the SCF/c-kit signaling axis in cancer, we generated a c-kit antagonist human antibody (NN2101) that binds to domain 2/3 of c-kit. This completely blocked the SCF-mediated phosphorylation of c-kit and inhibited TF-1 cell proliferation, erythroleukemia. In addition, the examination of binding affinity using surface plasmon resonance (SPR) assay showed that NN2101 can bind to c-kit of monkeys (KD = 2.92 × 10-10M), rats (KD = 1.68 × 10-6 M), mice (KD = 11.5 × 10-9 M), and humans (KD = 2.83 × 10-12 M). We showed that NN2101 does not cause antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity. The immunogenicity of NN2101 was similar to that of bevacizumab. Furthermore, the crystal structure of NN2101 Fab was determined and the structure of NN2101 Fab:c-kit complex was modeled. Structural information, as well as mutagenesis results, revealed that NN2101 can bind to the SCF-binding regions of c-kit. Collectively, we generated a c-kit neutralizing human antibody (NN2101) for the treatment of erythroleukemia and characterized its biophysical properties. NN2101 can potentially be used as a therapeutic antibody to treat different cancers.

2.
Arterioscler Thromb Vasc Biol ; 39(10): 2120-2131, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31434494

RESUMO

OBJECTIVE: Aberrant neovascularization is a leading cause of blindness in several eye diseases, including age-related macular degeneration and proliferative diabetic retinopathy. The identification of key regulators of pathological ocular neovascularization has been a subject of extensive research and great therapeutic interest. Here, we explored the previously unrecognized role of cKIT and its ligand, SCF (stem cell factor), in the pathological ocular neovascularization process. Approach and Results: Compared with normoxia, hypoxia, a crucial driver of neovascularization, caused cKIT to be highly upregulated in endothelial cells, which significantly enhanced the angiogenic response of endothelial cells to SCF. In murine models of pathological ocular neovascularization, such as oxygen-induced retinopathy and laser-induced choroidal neovascularization models, cKIT and SCF expression was significantly increased in ocular tissues, and blockade of cKIT and SCF using cKit mutant mice and anti-SCF neutralizing IgG substantially suppressed pathological ocular neovascularization. Mechanistically, SCF/cKIT signaling induced neovascularization through phosphorylation of glycogen synthase kinase-3ß and enhancement of the nuclear translocation of ß-catenin and the transcription of ß-catenin target genes related to angiogenesis. Inhibition of ß-catenin-mediated transcription using chemical inhibitors blocked SCF-induced in vitro angiogenesis in hypoxia, and injection of a ß-catenin agonist into cKit mutant mice with oxygen-induced retinopathy significantly enhanced pathological neovascularization in the retina. Conclusions; Our data reveal that SCF and cKIT are promising novel therapeutic targets for treating vision-threatening ocular neovascular diseases.

3.
Biomaterials ; 220: 119408, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31394431

RESUMO

Aminoacyl-tRNA synthetase (ARS)-interacting multifunctional protein 1 (AIMP1) enhances the expression of proinflammatory cytokines. In our previous study, we have shown that serum AIMP1 in patients with SLE was significantly higher than that of healthy controls. To address whether neutralization of AIMP1 could ameliorate nephritis in lupus-prone mice, we generated atializumab, a humanized antibody against AIMP1 and investigated its therapeutic efficacy. ELISA showed that serum AIMP1 at 23 weeks old was significantly higher than that at 13 weeks old in lupus-prone mice. Therefore, lupus-prone mice were randomly assigned to 5 groups (vehicle, methylprednisolone and 0.5, 2, and 5 mg/kg atializumab). After treatment, disease severity was assessed using a variety of phenotypes, including proteinuria, histological damages, renal deposition of immune-complex. In addition, serum cytokines, anti-dsDNA and IgG subclasses were determined. T cell subsets were analyzed using a fluorescence-activated cell sorter. Atializumab significantly diminished proteinuria, improved glomerular and tubular damages and reduced the renal deposition of immune-complexes. Moreover, atializumab significantly decreased serum interferon (IFN)-γ, interleukin (IL)-17A, and IL-6, whereas it increased serum IL-10. Similarly, atializumab reduced the numbers of TH1, TH2 and TH17 cells in a dose-dependent manner, while atializumab enhanced the number of regulatory T (Treg) cells. Furthermore, atializumab decreased not only splenic plasma cells and serum anti-dsDNA but also pathogenic IgG subclasses for nephritis. It suppressed NF-κB activation by inhibiting IκBα degradation in a dose-dependent manner in vitro. Atializumab alleviated nephritis by inhibiting autoreactive T, B, and plasma cells and decreasing NF-κB-related proinflammatory cytokines in lupus-prone mice. These results suggest that treatment targeting AIMP1 could be a novel and highly immune-modulating therapeutic strategy in lupus nephritis.

4.
Medicine (Baltimore) ; 98(23): e15885, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31169696

RESUMO

Duodenal neuroendocrine tumors (NETs) are rare, and risk factors associated with lymph node (LN) metastasis are still not well defined. The aim of this study was to investigate risk factors of LN metastasis in duodenal NETs based on the final histopathologic results and clinical follow-up data.This study included a total of 44 duodenal NETs in 38 patients who underwent endoscopic or surgical resection between January 2008 and December 2015. Diagnosis of duodenal NETs was confirmed based on immunohistochemical staining of chromogranin A, synaptophysin, and CD56; the clinicopathologic records were collected at the time of the initial diagnosis of duodenal NETs.Most duodenal NETs were small (≤1 cm in 33 tumors), World Health Organization (WHO) grade G1 (in 32 tumors), limited to the mucosa and/or submucosa (in 40 tumors), and located at the duodenal bulb (in 32 tumors). Of 44 tumors, lymphovascular invasion was present in 4 (9.1%), and among 38 patients, LN metastasis was detected in 4 (10.5%). LN metastases were significantly associated with the non-bulb location, tumor size >10 mm, tumor invasion into the muscularis propria or deeper, WHO grade G2, and lymphovascular invasion. During the mean follow-up period of 54.5 months (range, 24-123 months), recurrence occurred in 1 patient.Non-bulb location, tumor size >10 mm, invasion beyond the submucosa, WHO grade G2, and lymphovascular invasion are risk factors of LN metastasis in duodenal NETs. These findings can help clinicians choose the appropriate therapeutic modality for duodenal NETs.


Assuntos
Neoplasias Duodenais/patologia , Metástase Linfática/patologia , Tumores Neuroendócrinos/patologia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Invasividade Neoplásica , Recidiva Local de Neoplasia/patologia , Estudos Retrospectivos , Fatores de Risco , Carga Tumoral
5.
Biomed Res Int ; 2019: 7508240, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31236412

RESUMO

We investigated whether serum aminoacyl-tRNA synthetase-interacting multifunctional protein-1 (AIMP1) could predict severe cases of antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) based on the Birmingham vasculitis activity score (BVAS). Sixty-one patients with AAV were selected for inclusion from our prospective AAV cohort. AAV-specific indices and clinical manifestations were assessed, and laboratory tests were performed on the day of blood sampling. Patients with severe AAV were defined as those with a BVAS higher than the lower limit of the highest tertile of BVAS (BVAS ≥ 12). We measured serum AIMP1 levels of the stored serum samples. A total of 20 (32.8%) and 41 (67.2%) patients were classified as having severe and nonsevere AAV according to the cut-off of BVAS ≥ 12. Patients with severe AAV showed higher frequencies of general and renal manifestations, along with ANCA positivity, and exhibited a higher mean neutrophil count, erythrocyte sedimentation rate, and C-reactive protein levels, but lower mean haemoglobin and serum albumin levels than those with nonsevere AAV. The mean serum AIMP1 level in patients with severe AAV was significantly higher than that of patients with nonsevere AIMP1 (351.1 vs. 98.4 pg/mL, p = 0.006). Multivariate logistic regression analysis including variables showing significance in univariate analyses revealed that only serum AIMP1 exhibited a significant association with severe AAV (odds ratio 1.004, p = 0.031). When we set the optimal cut-off of serum AIMP1 for severe AAV to 50.28 pg/mL, patients with severe AAV more frequently had AIMP1 levels above the cut-off than those with nonsevere AAV (80.0% vs. 31.7%, relative risk 8.615, p < 0.001). The results from our study suggest that serum AIMP1 can be used to estimate the cross-sectional severe AAV population based on the BVAS.


Assuntos
Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos/sangue , Anticorpos Anticitoplasma de Neutrófilos/sangue , Biomarcadores/sangue , Citocinas/sangue , Proteínas de Neoplasias/sangue , Proteínas de Ligação a RNA/sangue , Adulto , Idoso , Aminoacil-tRNA Sintetases/genética , Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos/genética , Vasculite Associada a Anticorpo Anticitoplasma de Neutrófilos/patologia , Anticorpos Anticitoplasma de Neutrófilos/genética , Sedimentação Sanguínea , Estudos Transversais , Feminino , Humanos , Rim/metabolismo , Rim/patologia , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Estudos Prospectivos , Índice de Gravidade de Doença
6.
Int J Mol Sci ; 20(7)2019 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-30970566

RESUMO

The mammalian mediator complex subunit 28 (MED28) is overexpressed in a variety of cancers and it regulates cell migration/invasion and epithelial-mesenchymal transition. However, transcription factors that increase MED28 expression have not yet been identified. In this study, we performed a luciferase reporter assay to identify and characterize the prospective transcription factors, namely E2F transcription factor 1, nuclear respiratory factor 1, E-26 transforming sequence 1, and CCAAT/enhancer-binding protein ß, which increased MED28 expression. In addition, the release from the arrest at the G1-S or G2-M phase transition after cell cycle synchronization using thymidine or nocodazole, respectively, showed enhanced MED28 expression at the G1-S transition and mitosis. Furthermore, the overexpression of MED28 significantly decreased the duration of interphase and mitosis. Conversely, a knockdown of MED28 using si-RNA increased the duration of interphase and mitosis. Of note, the overexpression of MED28 significantly increased micronucleus and nuclear budding in HeLa cells. In addition, flow cytometry and fluorescence microscopy analyses showed that the overexpression of MED28 significantly increased aneuploid cells. Taken together, these results suggest that MED28 expression is increased by oncogenic transcription factors and its overexpression disturbs the cell cycle, which results in genomic instability and aneuploidy.


Assuntos
Instabilidade Genômica , Complexo Mediador/genética , Complexo Mediador/metabolismo , Fatores de Transcrição/metabolismo , Aneuploidia , Ciclo Celular/efeitos dos fármacos , Instabilidade Genômica/efeitos dos fármacos , Células HEK293 , Células HeLa , Humanos , Células MCF-7 , Nocodazol/farmacologia , Regiões Promotoras Genéticas , Timidina/farmacologia , Regulação para Cima
7.
Korean J Intern Med ; 2019 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-30879288

RESUMO

Background/Aims: Real-world, clinical practice data are lacking about sofosbuvir/ ribavirin (SOF/RBV) treatment of Korean patients with hepatitis C virus genotype 2 (HCV GT2) infection. This study investigated the efficacy and safety of SOF/RBV in Korean patients with HCV GT2 infection and clinical factors predicting sustained virological response 12 weeks (SVR12) after the end of SOF/RBV treatment. Methods: A total of 181 patients with HCV GT2 with/without cirrhosis were treated with SOF/RBV for 16/12 weeks. Rapid virological response (RVR) was defined as non-detectable HCV RNA at 4 weeks. Results: The RVR rate was 80.7% (146/181), the end of treatment response rate was 97.8% (177/181) and the SVR12 rate was 92.8% (168/181). Of eight patients with relapse, four did not achieve RVR. Three patients had a history of hepatocellular carcinoma (HCC). Multivariable analysis showed that RVR (p = 0.015) and no previous history of HCC (p = 0.007) were associated with SVR12. Factors significantly contributing to RVR included cirrhosis, creatinine concentration, and pre-treatment HCV RNA level. SVR12 rate was significantly higher in RVR (+) than RVR (-) patients (95.2% vs. 82.9%, p = 0.011) and also significantly higher in patients without than with a history of HCC (94.1% vs. 72.7%, p = 0.008). During treatment, 80/181 patients (44.2%) experienced mild to moderate adverse events, with 32 (17.7%) requiring RBV dose reductions due to anemia. Conclusions: SOF/RBV treatment was effective and tolerable in HCV GT2 patients. RVR and no previous history of HCC were positive predictors of SVR12.

8.
Sci Rep ; 9(1): 1656, 2019 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-30733541

RESUMO

Peroxisome proliferator-activated receptor (PPAR)-α/γ dual agonists have been developed to treat metabolic diseases; however, most of them exhibit side effects such as body weight gain and oedema. Therefore, we developed a novel PPARα/γ dual agonist that modulates glucose and lipid metabolism without adverse effects. We synthesised novel compounds composed of coumarine and chalcone, determined their crystal structures, and then examined their binding affinity toward PPARα/γ. We investigated the expression of PPARα and PPARγ target genes by chemicals in HepG2, differentiated 3T3-L1, and C2C12 cells. We examined the effect of chemicals on glucose and lipid metabolism in db/db mice. Only MD001 functions as a PPARα/γ dual agonist in vitro. MD001 increased the transcriptional activity of PPARα and PPARγ, resulting in enhanced expression of genes related to ß-oxidation and fatty acid and glucose uptake. MD001 significantly improved blood metabolic parameters, including triglycerides, free fatty acids, and glucose, in db/db mice. In addition, MD001 ameliorated hepatic steatosis by stimulating ß-oxidation in vitro and in vivo. Our results demonstrated the beneficial effects of the novel compound MD001 on glucose and lipid metabolism as a PPARα/γ dual agonist. Consequently, MD001 may show potential as a novel drug candidate for the treatment of metabolic disorders.

9.
J Nanobiotechnology ; 17(1): 19, 2019 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-30696428

RESUMO

BACKGROUND: Protein-based Cas9 in vivo gene editing therapeutics have practical limitations owing to their instability and low efficacy. To overcome these obstacles and improve stability, we designed a nanocarrier primarily consisting of lecithin that can efficiently target liver disease and encapsulate complexes of Cas9 with a single-stranded guide RNA (sgRNA) ribonucleoprotein (Cas9-RNP) through polymer fusion self-assembly. RESULTS: In this study, we optimized an sgRNA sequence specifically for dipeptidyl peptidase-4 gene (DPP-4) to modulate the function of glucagon-like peptide 1. We then injected our nanocarrier Cas9-RNP complexes directly into type 2 diabetes mellitus (T2DM) db/db mice, which disrupted the expression of DPP-4 gene in T2DM mice with remarkable efficacy. The decline in DPP-4 enzyme activity was also accompanied by normalized blood glucose levels, insulin response, and reduced liver and kidney damage. These outcomes were found to be similar to those of sitagliptin, the current chemical DPP-4 inhibition therapy drug which requires recurrent doses. CONCLUSIONS: Our results demonstrate that a nano-liposomal carrier system with therapeutic Cas9-RNP has great potential as a platform to improve genomic editing therapies for human liver diseases.


Assuntos
Sistemas CRISPR-Cas , Diabetes Mellitus Tipo 2/terapia , Dipeptidil Peptidase 4/genética , Sistemas de Liberação de Medicamentos , Terapia Genética/métodos , Lecitinas , Lipossomos , Animais , Glicemia/efeitos dos fármacos , Linhagem Celular , Dipeptidil Peptidase 4/metabolismo , Edição de Genes , Marcação de Genes , Peptídeo 1 Semelhante ao Glucagon/sangue , Humanos , Lecitinas/administração & dosagem , Lecitinas/química , Lipossomos/administração & dosagem , Lipossomos/química , Camundongos , Camundongos Knockout , RNA Guia/administração & dosagem , RNA Guia/química , RNA Guia/genética
10.
Korean J Gastroenterol ; 72(1): 28-32, 2018 Jul 25.
Artigo em Coreano | MEDLINE | ID: mdl-30049175

RESUMO

An inflammatory myofibroblastic tumor (IMT) is a rare disease that can occur in a variety of locations, including the lung, orbit, parotid, pleura, and stomach. Despite multiple reports in various organs, a duodenal IMT is rare with limited case reports. We encountered a case of a 49-year-old male with a duodenal IMT. The patient underwent a laparoscopic wedge resection under the impression of a duodenal mesenchymal tumor, such as gastrointestinal stromal tumor, but the final diagnosis was a duodenal IMT. The patient was treated successfully with an oral nonsteroidal anti-inflammatory drug for the residual lesions. He was free of recurrence during the 12 month follow-up period.


Assuntos
Neoplasias Duodenais/diagnóstico , Neoplasias de Tecido Muscular/diagnóstico , Actinas/metabolismo , Quinase do Linfoma Anaplásico/metabolismo , Neoplasias Duodenais/tratamento farmacológico , Neoplasias Duodenais/patologia , Endoscopia Gastrointestinal , Humanos , Masculino , Pessoa de Meia-Idade , Naproxeno/uso terapêutico , Neoplasias de Tecido Muscular/tratamento farmacológico , Neoplasias de Tecido Muscular/patologia , Tomografia Computadorizada por Raios X
11.
Saf Health Work ; 9(2): 224-231, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29928538

RESUMO

Background: Organizations are pursing complex and diverse aims to generate higher profits. Many workers experience high work intensity such as workload and work pressure in this organizational environment. Especially, psychological burden is a commonly used term in workplace of Republic of Korea. This study focused on defining the psychological burden from the perspective of occupational safety and health and tried to develop a scale for psychological burden. Methods: The 48 preliminary questionnaire items for psychological burden were prepared by a focus group interview with 16 workers through the Copenhagen Psychosocial Questionnaire II and Mindful Awareness Attention Scale. The preliminary items were surveyed with 572 workers, and exploratory factor analysis, confirmatory factor analysis, and correlation analysis were conducted for a new scale. Results: As a result of the exploratory factor analysis, five factors were extracted: organizational activity, human error, safety and health workload, work attitude, and negative self-management. These factors had significant correlations and reliability, and the stability of the model for validity was confirmed using confirmatory factor analysis. Conclusion: The developed scale for psychological burden can measure workers' psychological burden in relation to safety and health. Despite some limitations, this study has applicability in the workplace, given the relatively small-sized questionnaire.

12.
Clin Endosc ; 51(6): 587-590, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29764009

RESUMO

Schwannomas are uncommon neoplasms that arise from Schwann cells of the neural sheath. Gastrointestinal schwannomas are rare among mesenchymal tumors of the gastrointestinal tract, and only a few cases have been reported to date. Duodenal schwannomas are usually discovered incidentally and achieving a preoperative diagnosis is difficult. Schwannomas can be distinguished from other subepithelial tumors on endoscopic ultrasonography; however, any typical endosonographic features of duodenal schwannomas have not been reported due to the rarity of these tumors. Immunohistochemistry is essential to distinguish schwannomas from gastrointestinal stromal tumors and leiomyomas. We report a case of duodenal schwannoma found incidentally during a health check-up endoscopy. On endoscopic ultrasonography, this tumor was suspected as a gastrointestinal stromal tumor; therefore, the patient underwent laparoscopic wedge resection of the tumor. Histopathology and immunohistochemistry confirmed that the duodenal lesion was a benign schwannoma.

13.
Oncol Lett ; 15(3): 3147-3154, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29435049

RESUMO

The regulation of gene expression by transcription factors serves a critical function in cell proliferation. Zinc-finger protein 224 (ZNF224), a Krüppel-associated-box-containing zinc finger protein, is known to serve a crucial function in integrating the transcriptional co-factors that activate transcriptional regulation pathways in the cell. A previous study demonstrated that ZNF224 enhances cell proliferation by downregulating the expression of p21 and p53. The present study identified mediator complex subunit 28 (MED28) as a potential binding partner for ZNF224; this was confirmed by co-immunoprecipitation and a surface plasmon resonance assay. Additionally, the KRAB domain at the N-terminal of ZNF224 interacts with the MED domain of MED28. Bimolecular fluorescence complementation analysis revealed that ZNF224 associates with MED28 in the nucleus. In addition, ZNF224 was rapidly degraded upon treatment with the DNA-damaging agent camptothecin (CPT). Transient overexpression of MED28 inhibited the CPT-mediated degradation of ZNF224, resulting in increased colony formation by MCF-7 cells. The molecular mechanisms that underlie the biological outcomes of MED28 expression have not yet been fully elucidated. The present study provides molecular evidence for the function of ZNF224 and MED28 in the DNA-damage response.

14.
Clin Exp Rheumatol ; 36(4): 533-539, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29352840

RESUMO

OBJECTIVES: Secreted aminoacyl-tRNA synthetase-interacting multifunctional protein-1 (AIMP1) has been reported to have pro-inflammatory properties. The aim of this study was to evaluate the clinical significance of serum AIMP1 in patients with systemic lupus erythematosus (SLE). METHODS: Serum levels of AIMP1 were measured in 160 patients with SLE using a human AIMP1 ELISA kit. Eighty patients were classified as active SLE (SLEDAI-2K ≥ 5), and 80 patients were classified as stable SLE. Correlation between serum AIMP1, SLE disease activity index-2000 (SLEDAI-2K), and laboratory variables related to disease activity or inflammatory burdens were assessed using Pearson's correlation analysis. The optimal cut-off value for serum AIMP1 to predict active SLE was estimated by using a receiver operator characteristic curve, and logistic regression analysis was used to compare the odds ratios (ORs) of laboratory variables in predicting active SLE. RESULTS: The median serum AIMP1 was higher in patients with active SLE than those with stable SLE (8.0 vs. 6.5 ng/ml, p<0.001). Serum AIMP1 demonstrated correlation with SLEDAI-2K and laboratory variables related to disease activity or inflammatory burdens. The optimal cut-off AIMP1 to predict active SLE was 10.09. Multivariate logistic regression analysis including conventional laboratory variables demonstrated that serum AIMP1 ≥10.09 ng/ml (OR 3.919, 95% confidence interval 1.223-12.564, p=0.022) was useful in predicting active SLE. CONCLUSIONS: Serum levels of AIMP1 were associated with disease activity of SLE and could predict active SLE based on SLEDAI-2K.


Assuntos
Citocinas/sangue , Lúpus Eritematoso Sistêmico/sangue , Proteínas de Neoplasias/sangue , Proteínas de Ligação a RNA/sangue , Adulto , Biomarcadores , Feminino , Humanos , Modelos Logísticos , Lúpus Eritematoso Sistêmico/diagnóstico , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença
15.
Int J Mol Sci ; 18(8)2017 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-28813021

RESUMO

The multipotency and anti-inflammatory effects of mesenchymal stem cells (MSCs) make them attractive for cell therapy in regenerative medicine. A large number of MSCs is required for efficient therapy owing to the low homing efficiency of MSCs to target sites. Furthermore, owing to limitations in obtaining sufficient amounts of MSCs, in vitro expansion of MSCs that preserves their differentiation and proliferative potential is essential. The animal factor included in culture media also limits clinical application. In this study, adipose-derived MSCs showed a significantly higher proliferation rate in STK2, a chemically-defined medium, than in DMEM/FBS. The expression of MSC surface markers was increased in the culture using STK2 compared to that using DMEM/FBS. Tri-lineage differentiation analyses showed that MSCs cultured in STK2 were superior to those cultured in DMEM/FBS. In addition, MSCs cultured in STK2 showed a reduced senescence rate, small and homogenous cell size, and were more genetically stable compared to those cultured in DMEM/FBS. Furthermore, secretome analysis showed that the expression of factors related to proliferation/migration, anti-inflammation, and differentiation were increased in STK2 culture medium compared to DMEM/FBS. Taken together, these results suggest that culture using STK2 medium offers many advantages through which it is possible to obtain safer, superior, and larger numbers of MSCs.


Assuntos
Adipócitos/citologia , Adipócitos/metabolismo , Técnicas de Cultura de Células/métodos , Meios de Cultura/química , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Humanos
16.
Auris Nasus Larynx ; 44(2): 199-204, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27269133

RESUMO

OBJECTIVE: An inflammatory-immunological marker, C-reactive protein (CRP), neutrophil-to-lymphocyte ratio (NLR), was evaluated as a predictive marker of advanced head and neck cancer patients receiving chemoradiotherapy. METHODS: This study included 104 patients with treatment-naïve head and neck cancer who underwent definitive chemoradiotherapy. An inflammatory marker was measured at baseline and after 1 month of treatment. Univariate and multivariate analyses using Cox proportional hazards model were used to identify predictors of progression-free survival (PFS) and overall survival (OS). RESULTS: A univariate analysis revealed that T,N-stage, the pre- and posttreatment NLRs were significant predictors of progression after the chemoradiotherapy. However, the posttreatment NLR remained an independent predictor of PFS in the multivariate analysis (HR=2.23, 95% CI 1.15-2.321; P=0.001). A high posttreatment NLR was significantly associated with an increased risk of death (HR=1.87, 95% CI 0.89-3.31; P=0.037). CONCLUSION: A high posttreatment NLR is associated with poor prognostic factor. An early reduction in the NLR after treatment may indicate survival improvement in the patients.


Assuntos
Antineoplásicos/uso terapêutico , Proteína C-Reativa/metabolismo , Carcinoma de Células Escamosas/terapia , Quimiorradioterapia , Cisplatino/uso terapêutico , Neoplasias de Cabeça e Pescoço/terapia , Linfócitos/citologia , Neutrófilos/citologia , Adulto , Idoso , Carcinoma de Células Escamosas/sangue , Carcinoma de Células Escamosas/metabolismo , Feminino , Neoplasias de Cabeça e Pescoço/sangue , Neoplasias de Cabeça e Pescoço/metabolismo , Humanos , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Modelos de Riscos Proporcionais , Carcinoma de Células Escamosas de Cabeça e Pescoço
17.
Neuroreport ; 28(2): 82-86, 2017 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-27906773

RESUMO

Neuroinflammation has been raised as a candidate of unifying pathogenesis and a target of a disease-modifying strategy for Alzheimer's disease (AD). Aminoacyl-tRNA synthetase complex (ARS)-interacting multifunctional protein 1 (AIMP1) is a cytokine that is known to amplify the actions of tumor necrosis factor-α and to be involved in microglial activation and neuronal death. In this respect, AIMP1 could be a plausible target for the treatment of AD. Therefore, we aimed to examine whether anti-AIMP1 antibody could exert therapeutic effects against cognitive impairment using 3xTg-AD mice. Through the passive avoidance test, we found that an intraperitoneal injection of anti-AIMP1 antibody over 4 weeks was effective in protecting memory function in 3xTg-AD mice (16 weeks old). In addition, to address the translational implications of AIMP1, we measured blood AIMP1 levels in patients with AD (n=22), mild cognitive impairment (n=25), and normal cognition (n=23). Blood AIMP1 levels were associated negatively with global cognitive function and were significantly higher in individuals with a higher degree of medial temporal lobe atrophy, which is one of the representative clinical markers of AD. Our results suggested a possible association of AIMP1 with AD pathogenesis, as well as the potential of the anti-AIMP1 antibody as a novel therapeutic option for AD.


Assuntos
Doença de Alzheimer/complicações , Transtornos Cognitivos/metabolismo , Transtornos Cognitivos/prevenção & controle , Citocinas/metabolismo , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/sangue , Doença de Alzheimer/genética , Doença de Alzheimer/patologia , Precursor de Proteína beta-Amiloide/genética , Animais , Anticorpos/uso terapêutico , Aprendizagem da Esquiva/efeitos dos fármacos , Encéfalo/diagnóstico por imagem , Citocinas/imunologia , Modelos Animais de Doenças , Feminino , Humanos , Imagem por Ressonância Magnética , Masculino , Transtornos da Memória/etiologia , Entrevista Psiquiátrica Padronizada , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Pessoa de Meia-Idade , Mutação/genética , Presenilina-1/genética , Teste de Desempenho do Rota-Rod , Proteínas tau/genética
19.
Biochem Biophys Res Commun ; 478(4): 1682-7, 2016 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-27596970

RESUMO

Tauroursodeoxycholic acid (TUDCA) is known to prevent apoptosis through the Bax pathway and to promote neovascularization by enhancing the mobilization of stem cells, their differentiation. This study was performed to investigate the effect of TUDCA on erythropoiesis in hematopoietic stem cells (HSCs). Since erythropoiesis of CD34(+) HSCs is divided into four phases, the total cell number, viable cell number, cell viability, cell morphology, and expressed erythroid markers in each phase were examined. The number of viable control cells and their viability did not differ from those of the TUDCA-treated cells in phase I and II. However, TUDCA increased cell viability compared to the control in phases III and IV. Cell distribution differed that the immature erythroid cell number was higher for the TUDCA-treated cells than for the control cells until phase III, but all developed into RBCs in the last. The final RBC number and viability was significantly higher in TUDCA-treated cells compared to the control cells. Taken together, we suggest that TUDCA addition to cell cultures for artificial RBC production could be used as a new protocol for improving the viability of RBCs.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Eritrócitos/efeitos dos fármacos , Células Eritroides/efeitos dos fármacos , Células-Tronco Hematopoéticas/efeitos dos fármacos , Ácido Tauroquenodesoxicólico/farmacologia , Adulto , Idoso , Antígenos CD34/metabolismo , Biomarcadores/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Colagogos e Coleréticos/farmacologia , Contagem de Eritrócitos , Eritrócitos/citologia , Eritrócitos/metabolismo , Células Eritroides/citologia , Células Eritroides/metabolismo , Eritropoese/efeitos dos fármacos , Feminino , Citometria de Fluxo , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/metabolismo , Humanos , Imunofenotipagem , Masculino , Pessoa de Meia-Idade , Fatores de Tempo
20.
Int J Mol Sci ; 17(9)2016 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-27563882

RESUMO

Hypoxia has diverse stimulatory effects on human adipose-derived stem cells (ASCs). In the present study, we investigated whether hypoxic culture conditions (2% O2) suppress spontaneous mineralization and osteogenic differentiation of ASCs. We also investigated signaling pathways and molecular mechanisms involved in this process. We found that hypoxia suppressed spontaneous mineralization and osteogenic differentiation of ASCs, and up-regulated mRNA and protein expression of Insulin-like growth factor binding proteins (IGFBPs) in ASCs. Although treatment with recombinant IGFBPs did not affect osteogenic differentiation of ASCs, siRNA-mediated inhibition of IGFBP3 attenuated hypoxia-suppressed osteogenic differentiation of ASCs. In contrast, overexpression of IGFBP3 via lentiviral vectors inhibited ASC osteogenic differentiation. These results indicate that hypoxia suppresses spontaneous mineralization and osteogenic differentiation of ASCs via intracellular IGFBP3 up-regulation. We determined that reactive oxygen species (ROS) generation followed by activation of the MAPK and PI3K/Akt pathways play pivotal roles in IGFBP3 expression under hypoxia. For example, ROS scavengers and inhibitors for MAPK and PI3K/Akt pathways attenuated the hypoxia-induced IGFBP3 expression. Inhibition of Elk1 and NF-κB through siRNA transfection also led to down-regulation of IGFBP3 mRNA expression. We next addressed the proliferative potential of ASCs with overexpressed IGFBP3, but IGFBP3 overexpression reduced the proliferation of ASCs. In addition, hypoxia reduced the osteogenic differentiation of bone marrow-derived clonal mesenchymal stem cells. Collectively, our results indicate that hypoxia suppresses the osteogenic differentiation of mesenchymal stem cells via IGFBP3 up-regulation.


Assuntos
Diferenciação Celular/fisiologia , Hipóxia Celular/fisiologia , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Osteogênese/fisiologia , Animais , Diferenciação Celular/genética , Hipóxia Celular/genética , Proliferação de Células/genética , Proliferação de Células/fisiologia , Células Cultivadas , Humanos , Imunoprecipitação , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Camundongos , Osteogênese/genética , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
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