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1.
Clin Med Insights Endocrinol Diabetes ; 14: 11795514211040539, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34602832

RESUMO

Background: The safe method of instructing insulin dose reduction in combination with SGLT2 inhibitors, dapagliflozin for patients with type 1 diabetes mellitus has not been clarified. In this study, we conducted a stratified, 2-arm, parallel comparative study with the primary endpoint of decreasing the frequency of hypoglycemia by instructing basal insulin dose reduction. Methods: The study has a multicenter, open-label, 2-arm design; 60 type 1 diabetes mellitus patients are being recruited from 7 hospitals. Study subjects have been stratified into 2 groups based on the ratio of basal insulin daily dose (Basal) to total daily insulin dose (TDD). The subjects whose Basal/TDD ratio is <0.4 are instructed not to reduce Basal but to reduce bolus insulin dose by 10% (group A), and subjects with a Basal/TDD ratio >0.4 will be instructed to reduce Basal by 10% (group B). The primary outcome is the daily frequency of hypoglycemia during the intervention period (SGLT2 inhibitor administration), as determined by self-monitoring of blood glucose. We aimed to confirm a greater reduction in frequency of hypoglycemia in group B (reduced Basal), than in group A (non-reduction of Basal and reduced insulin effect levels by 10%). Baseline hypoglycemia was set at 7 ± 6 times/month. The minimum sample size required to achieve a significance of .05 for a 1-sided t-test with a statistical power at 80% is determined. When the sample size is 26 patients in 1 group, the percentage increase in hypoglycemia exceeds 60%, and the sample size is considered sufficient. Discussion: In this pilot study, we assumed that, given a sufficient Basal, hypoglycemia would be more frequent in patients with type 1 diabetes when combined with SGLT2 inhibitors, provided the Basal was not reduced.

2.
J Am Chem Soc ; 143(33): 13044-13055, 2021 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-34387474

RESUMO

Reprogramming known medicines for a novel target with activity and selectivity over the canonical target is challenging. By studying the binding interactions between RNA folds and known small-molecule medicines and mining the resultant dataset across human RNAs, we identified that Dovitinib, a receptor tyrosine kinase (RTK) inhibitor, binds the precursor to microRNA-21 (pre-miR-21). Dovitinib was rationally reprogrammed for pre-miR-21 by using it as an RNA recognition element in a chimeric compound that also recruits RNase L to induce the RNA's catalytic degradation. By enhancing the inherent RNA-targeting activity and decreasing potency against canonical RTK protein targets in cells, the chimera shifted selectivity for pre-miR-21 by 2500-fold, alleviating disease progression in mouse models of triple-negative breast cancer and Alport Syndrome, both caused by miR-21 overexpression. Thus, targeted degradation can dramatically improve selectivity even across different biomolecules, i.e., protein versus RNA.

3.
Artigo em Inglês | MEDLINE | ID: mdl-34203155

RESUMO

Objectives: We verified the clinical usefulness of an approach method in which a physician gives simple salt reduction instructions during outpatient visits to patients with type 2 diabetes. Methods: This study was an open-blind, randomized controlled trial. Subjects were outpatients with type 2 diabetes whose estimated salt intake using spot morning urine sample exceeded the target of salt intake. The control group (CG) was notified only of the current salt intake, whereas the intervention group (IG) was given the brief salt reduction instruction by a physician in addition to the information regarding their current salt intake. Results: The change in estimated salt intake was -0.6 g (from 10.1 to 9.5 g, p = 0.029) in the CG after 8 weeks, and -0.9 g (from 10.1 to 9.2 g, p = 0.001) in the IG, although there were no significant differences between them (p = 0.47). After 24 weeks, both groups no longer differed significantly from the baseline. In addition, multivariate linear regression analyses indicated that high salt intake and low estimated glomerular filtration rate at baseline were significantly associated with salt reduction after 8 weeks. Conclusions: Salt-reducing effects were observed after 8 weeks in both the IG and CG, but no significant difference was observed. Moreover, patients with high salt intake and renal disfunction may be more effective in accepting salt reduction instructions. Making patients aware of the importance of salt reduction through a physician is effective for continuous salt reduction, and it is important to continue regular and repetitive guidance.


Assuntos
Diabetes Mellitus Tipo 2 , Hipertensão , Médicos , Diabetes Mellitus Tipo 2/prevenção & controle , Comportamento Alimentar , Taxa de Filtração Glomerular , Humanos , Cloreto de Sódio na Dieta
4.
Genes Cells ; 26(8): 611-626, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34081835

RESUMO

Serum/glucocorticoid-regulated kinase 1 (SGK1) is predominantly expressed in endothelial cells of mouse embryos, and Sgk1 null mice show embryonic lethality due to impaired vascular formation. However, how the SGK1 expression is controlled in developing vasculature remains unknown. In this study, we first identified a proximal endothelial enhancer through lacZ reporter mouse analyses. The mouse Sgk1 proximal enhancer was narrowed down to the 5' region of the major transcription initiation site, while a human corresponding region possessed relatively weak activity. We then searched for distal enhancer candidates using in silico analyses of publicly available databases for DNase accessibility, RNA polymerase association and chromatin modification. A region approximately 500 kb distant from the human SGK1 gene was conserved in the mouse, and the mouse and human genomic fragments drove transcription restricted to embryonic endothelial cells. Minimal fragments of both proximal and distal enhancers had consensus binding elements for the ETS transcription factors, which were essential for the responsiveness to ERG, FLI1 and ETS1 proteins in luciferase assays and the endothelial lacZ reporter expression in mouse embryos. These results suggest that endothelial SGK1 expression in embryonic vasculature is maintained through at least two ETS-regulated enhancers located in the proximal and distal regions.


Assuntos
Endotélio Vascular/metabolismo , Elementos Facilitadores Genéticos , Proteínas Imediatamente Precoces/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Animais , Cromatina/metabolismo , RNA Polimerases Dirigidas por DNA/metabolismo , Células Endoteliais/metabolismo , Endotélio Vascular/embriologia , Células HEK293 , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Proteínas Imediatamente Precoces/genética , Camundongos , Proteínas Oncogênicas/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteína Proto-Oncogênica c-ets-1/metabolismo , Proteína Proto-Oncogênica c-fli-1/metabolismo , Sítio de Iniciação de Transcrição , Regulador Transcricional ERG/metabolismo
5.
Int Endod J ; 54(10): 1902-1914, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34096634

RESUMO

AIM: To evaluate the dental pulp response to a novel mineral trioxide aggregate containing phosphorylated pullulan (MTAPPL) in rats after direct pulp capping. METHODS: Ninety-six cavities were prepared in the maxillary first molars of 56 male Wistar rats. The dental pulps were intentionally exposed and randomly divided into four groups according to the application of pulp capping materials: MTAPPL; phosphorylated pullulan (PPL); a conventional MTA (Nex-Cem MTA, NCMTA; positive control); and Super-Bond (SB; negative control). All cavities were restored with SB and observed for pulpal responses at 1-, 3-, 7- and 28-day intervals using a histological scoring system. Statistical analysis was performed using Kruskal-Wallis and Mann-Whitney U-test with Bonferroni's correction, and the level of significance was set at 0.05. DMP1 and CD34 antigen were used to evaluate odontoblast differentiation and pulpal vascularization, respectively. RESULTS: On day 1, mild inflammatory cells were present in MTAPPL and NCMTA groups; fewer inflammatory cells were present in the PPL, whereas SB was associated with a mild-to-moderate inflammatory response. A significant difference was observed between PPL and SB (p < .05). No mineralized tissue deposition was observed. On day 3, moderate-to-severe inflammatory cells were present in PPL and SB, whereas MTAPPL and NCMTA had a mild inflammatory response. Initial mineralized tissue deposition was observed in the NCMTA, MTAPPL and SB. A significant difference was observed between MTAPPL and PPL (p < .05). On day 7, a thin layer of mineralized tissue was observed in all tested groups with no or mild inflammatory response. On day 28, no inflammatory response was observed in MTAPPL, whereas NCMTA, PPL and SB had mild inflammatory responses. A significant difference was observed between MTAPPL and SB (p < .05). Complete mineralized tissue barrier formation was observed in MTAPPL, NCMTA and PPL with no significant difference (p > .05). SB exhibited incomplete mineralized tissue barriers, significantly different from NCMTA, MTAPPL and PPL (p < .05). The staining with CD34 was positive in all the groups on all observation days. CONCLUSION: The favourable pulpal responses and induction of mineralized tissue formation associated with MTAPPL indicate its potential application as a direct pulp capping material.


Assuntos
Capeamento da Polpa Dentária , Agentes de Capeamento da Polpa Dentária e Pulpectomia , Compostos de Alumínio , Animais , Compostos de Cálcio , Polpa Dentária , Combinação de Medicamentos , Glucanos , Masculino , Dente Molar , Óxidos , Agentes de Capeamento da Polpa Dentária e Pulpectomia/uso terapêutico , Ratos , Ratos Wistar , Silicatos
6.
Case Rep Med ; 2021: 5568978, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33883999

RESUMO

The spontaneous rupture of a pheochromocytoma is rare and can be potentially fatal. We report a case of a tumor size reduction of a ruptured pheochromocytoma after transcatheter arterial embolization (TAE). A 60-year-old Japanese woman was referred to the emergency department of another hospital with a sudden onset of left lateral pain. Computed tomography of the abdomen revealed adrenal hemorrhage with a 5.7 cm adrenal mass, and she was transferred to our hospital for treatment. Considering that she had marked hypertension (193/115 mmHg), we made a provisional diagnosis of left lateral pain due to a ruptured pheochromocytoma. She underwent TAE, and the hemorrhage was successfully controlled. She was started on oral doxazosin for hypertension. The dose of doxazosin was increased to the extent that orthostatic hypotension did not develop, and blood pressure was well controlled. After discharge, the tumor size gradually decreased to approximately 1.0 cm within six months. Six months after TAE, elective laparoscopic surgery was performed, and the diagnosis was confirmed by histopathology. We observed a decrease in the size of the ruptured pheochromocytoma after TAE. To reduce the risk of laparoscopic adrenal surgery, it may be useful to monitor the size of a ruptured pheochromocytoma after TAE before deciding the surgery time.

7.
J Arrhythm ; 37(2): 438-444, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33850586

RESUMO

Background: Early detection of cardiac involvement in patients with sarcoidosis is important but currently unresolved. The aim of this study was to elucidate the utility of frequency domain microvolt T-wave alternans (TWA), signal-averaged ECG (SAECG), and heart rate turbulence (HRT) using 24-hour Holter ECG for detecting cardiac involvement in patients with pulmonary sarcoidosis. Methods: This study consisted of consecutive 40 pulmonary sarcoidosis patients (11 males, 62 ± 13 years) who underwent 24-hour Holter monitoring with and without cardiac involvement. All patients underwent frequency domain TWA, SAECG, and HRT using 24-hour Holter monitoring. Patients with atrial fibrillation pacing or wide QRS electrocardiogram were excluded. Results: After 14 patients were excluded, a total of 26 patients (six males, 59 ± 14 years) were evaluated. Seven patients had cardiac involvement (cardiac sarcoidosis [CS] group). On the Holter SAECG, duration of low-amplitude signals <40 µV in the terminal filtered QRS complex (LAS40) was significantly higher, and root mean square voltage of the terminal 40 ms of the filtered QRS complex (RMS40) was significantly lower in the CS group compared with the non-CS group (LAS40: 61.4 ± 35.9 vs 37.6 ± 9.2 ms; P = .018, RMS40: 11.4 ± 10.3 vs 23.6 ± 13.2 ms; P = .023). Prevalence of positive late potential (LP) was also significantly higher in the CS group than that in the non-CS group (85.7% vs 31.5%; P = .026). The sensitivity, specificity, positive, and negative predictive values of LP for identifying patients with cardiac involvement were 85.7%, 68.4%, 50.0%, and 92.8%, respectively. Conclusion: Holter SAECG may be useful for detecting cardiac involvement in patients with pulmonary sarcoidosis.

8.
Biochem Biophys Res Commun ; 544: 15-21, 2021 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-33516877

RESUMO

Hepatocellular carcinoma (HCC) is one of the most common human malignant tumors. It is known that in the cells of many cancers, including HCC, nuclear translocation and accumulation of YB-1 often indicates a poor prognosis. This nuclear translocation is induced by genotoxic stress resulting from administration of anticancer agents. Accumulation of YB-1 in the nucleus induces the expression of many genes related to cancer aggressiveness. Therefore, compounds capable of inhibiting anticancer drug-induced YB-1 nuclear translocation without cytotoxicity will be a powerful tool for cancer chemotherapy. In the present study, we found that indirubin derivative, 7-hydroxyindirubin strongly inhibited the actinomycin D-induced nuclear translocation of YB-1 more efficiently without showing cytotoxicity in HepG2, a human HCC cells. The compound successfully suppressed the nuclear YB-1-mediated expression of genes such as MDR1, MVP, EGFR, and CXCR4, which are known to disturb cancer treatment. 7-Hydroxyindirubin also increased the susceptibility of drug-resistant HepG2 cells to ActD. It was also demonstrated that 7-hydroxyindirubin inhibits the nuclear translocation of YB-1 with or without phosphorylation at the Ser102 residue.


Assuntos
Carcinoma Hepatocelular/tratamento farmacológico , Núcleo Celular/metabolismo , Neoplasias Hepáticas/tratamento farmacológico , Proteína 1 de Ligação a Y-Box/metabolismo , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/farmacologia , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Células Hep G2 , Humanos , Indóis/química , Indóis/farmacologia , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Transporte Proteico , Receptores CXCR4/metabolismo
9.
Hypertens Res ; 44(3): 348-354, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33288879

RESUMO

Our previous study showed that the morning systolic blood pressure target should be <120 mmHg to prevent the onset or progression of diabetic nephropathy in patients with type 2 diabetes. In this study, we examined the prognostic values of home and clinical blood pressure for first cardiovascular events in the same cohort. Morning and evening home blood pressure measurements were obtained in triplicate for 14 consecutive days from the beginning of the study in a retrospective cohort of 1081 type 2 diabetes patients (44.5% women; median age 66.0 years) with no history of macrovascular complications. The first major cardiovascular event was the primary endpoint; the risk was examined by the Cox proportional hazards model. After a mean follow-up of 6.63 years, first-time cardiovascular events occurred in 119 patients (incidence, 16.6/1000 patient-years). Baseline morning systolic blood pressure (hazard ratio: 1.14, 95% CI 1.01-1.28) significantly predicted cardiovascular events, whereas clinical blood pressure did not. The adjusted hazard ratio (95% CI) for the incidence of cardiovascular events in patients with morning systolic blood pressure ≥135 mmHg tended to be higher than that in those with morning systolic blood pressure <125 mmHg [1.67 (0.94-2.97)]. Elevated home blood pressure measurement is a predictor of future cardiovascular events in type 2 diabetes patients and may be superior to clinical blood pressure measurement in this regard.

10.
Exp Cell Res ; 398(2): 112416, 2021 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-33307020

RESUMO

Idiopathic pulmonary fibrosis (IPF), a progressive disorder of unknown etiology, is characterized by pathological lung fibroblast activation and proliferation resulting in abnormal deposition of extracellular matrix proteins within the lung parenchyma. The pathophysiological roles of exosomal microRNAs in pulmonary fibrosis remain unclear; therefore, we aimed to identify and characterize fibrosis-responsive exosomal microRNAs. We used microRNA array analysis and profiled the expression of exosome-derived miRNA in sera of C57BL/6 mice exhibiting bleomycin-induced pulmonary fibrosis. The effect of microRNAs potentially involved in fibrosis was then evaluated in vivo and in vitro. The expression of exosomal microRNA-16 was increased by up to 8.0-fold on day 14 in bleomycin-treated mice, compared to vehicle-treated mice. MicroRNA-16 mimic administration on day 14 after bleomycin challenge ameliorated pulmonary fibrosis and suppressed lung and serum expression of secreted protein acidic and rich in cysteine (SPARC). Pretreatment of human lung fibroblasts with the microRNA-16 mimic decreased the expression of rapamycin-insensitive companion of mTOR (Rictor) and TGF-ß1-induced expression of SPARC. This is the first study reporting the anti-fibrotic properties of microRNA-16 and demonstrating that these effects occur via the mTORC2 pathway. These findings support that microRNA-16 may be a promising therapeutic target for IPF.


Assuntos
Fibrose Pulmonar Idiopática/metabolismo , Alvo Mecanístico do Complexo 2 de Rapamicina/metabolismo , MicroRNAs/metabolismo , Osteonectina/metabolismo , Proteína Companheira de mTOR Insensível à Rapamicina/metabolismo , Animais , Exossomos/metabolismo , Fibrose Pulmonar Idiopática/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL
11.
J Nippon Med Sch ; 88(4): 326-334, 2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32999175

RESUMO

BACKGROUND: Measuring lung compliance is useful for evaluating progression of interstitial lung disease (ILD), because reduced lung compliance due to fibrosis progression is the main cause of decreased vital capacity. However, because insertion of a balloon into the esophagus is invasive, lung compliance is rarely measured. A recently developed method uses fingertip photoplethysmography to estimate intrathoracic pressure. This method non-invasively measures lung dynamic compliance (Cdyn) by simultaneously measuring tidal volume. We evaluated the efficacy of this method in assessing ILD. METHODS: This single-center, cross-sectional, observational study evaluated the efficacy of this method in patients with ILD and healthy controls. The primary outcome was estimated Cdyn (eCdyn), as determined with this method. We also evaluated baseline characteristics that are potential confounding factors for eCdyn. RESULTS: Median eCdyn was significantly lower in the ILD group (n = 14) than in the control group (n = 49) (0.122 vs. 0.183; P = 0.011). In univariate regression analysis, eCdyn was significantly correlated with height, weight, forced vital capacity, forced expiratory volume in 1 second, diffusing capacity for carbon monoxide, and usual interstitial pneumonia. In multivariate regression analysis, weight (ß = 0.49, P = 0.011) and usual interstitial pneumonia (ß = 0.52, P = 0.007) were significantly correlated with eCdyn. CONCLUSIONS: Using photoplethysmography, we noted a significant reduction in Cdyn in patients with ILD. This novel non-invasive method is a promising tool for evaluating fibrosis progression in ILD.

12.
J Digit Imaging ; 33(6): 1543-1553, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33025166

RESUMO

Temporal subtraction (TS) technique calculates a subtraction image between a pair of registered images acquired from the same patient at different times. Previous studies have shown that TS is effective for visualizing pathological changes over time; therefore, TS should be a useful tool for radiologists. However, artifacts caused by partial volume effects degrade the quality of thick-slice subtraction images, even with accurate image registration. Here, we propose a subtraction method for reducing artifacts in thick-slice images and discuss its implementation in high-speed processing. The proposed method is based on voxel matching, which reduces artifacts by considering gaps in discretized positions of two images in subtraction calculations. There are two different features between the proposed method and conventional voxel matching: (1) the size of a searching region to reduce artifacts is determined based on discretized position gaps between images and (2) the searching region is set on both images for symmetrical subtraction. The proposed method is implemented by adopting an accelerated subtraction calculation method that exploit the nature of liner interpolation for calculating the signal value at a point among discretized positions. We quantitatively evaluated the proposed method using synthetic data and qualitatively using clinical data interpreted by radiologists. The evaluation showed that the proposed method was superior to conventional methods. Moreover, the processing speed using the proposed method was almost unchanged from that of the conventional methods. The results indicate that the proposed method can improve the quality of subtraction images acquired from thick-slice images.


Assuntos
Tomografia Computadorizada por Raios X , Algoritmos , Artefatos , Humanos , Radiologistas , Técnica de Subtração
13.
Materials (Basel) ; 13(21)2020 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-33113885

RESUMO

This study aimed to evaluate the bonding performance of a new one-step self-etching adhesive system containing a novel hydrophilic amide monomer. Clearfil Universal Bond Quick (CUB) and Clearfil Megabond 2 (CMB) were used as the one-step and two-step adhesive systems, respectively. Flat dentin surfaces of human premolars were exposed using #600 SiC (silicon carbide) and bonded with the respective adhesives of each system. The teeth were sectioned to obtain beams (1 mm × 1 mm) after 24 h of water storage. The mean bond strength and standard deviations (MPa) on an occlusal surface were as follows: CUB: 45.9 ± 19.7 and CMB: 67.9 ± 25.3. The values for cervical ones were CUB: 56.0 ± 20.3 and CMB: 67.6 ± 16.0, respectively. In both conditions, the microtensile bond strength (µTBS) value was lower than that of CMB. As seen during the microscopic observation, no adhesive failure was observed after µTBS testing because CUB formed a firm and tight adhesive interface.

14.
Chem Soc Rev ; 49(19): 7167-7199, 2020 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-32975549

RESUMO

Targeting RNAs with small molecules represents a new frontier in drug discovery and development. The rich structural diversity of folded RNAs offers a nearly unlimited reservoir of targets for small molecules to bind, similar to small molecule occupancy of protein binding pockets, thus creating the potential to modulate human biology. Although the bacterial ribosome has historically been the most well exploited RNA target, advances in RNA sequencing technologies and a growing understanding of RNA structure have led to an explosion of interest in the direct targeting of human pathological RNAs. This review highlights recent advances in this area, with a focus on the design of small molecule probes that selectively engage structures within disease-causing RNAs, with micromolar to nanomolar affinity. Additionally, we explore emerging RNA-target strategies, such as bleomycin A5 conjugates and ribonuclease targeting chimeras (RIBOTACs), that allow for the targeted degradation of RNAs with impressive potency and selectivity. The compounds discussed in this review have proven efficacious in human cell lines, patient-derived cells, and pre-clinical animal models, with one compound currently undergoing a Phase II clinical trial and another that recently garnerd FDA-approval, indicating a bright future for targeted small molecule therapeutics that affect RNA function.


Assuntos
MicroRNAs/efeitos dos fármacos , Bibliotecas de Moléculas Pequenas/farmacologia , Animais , Sistemas de Liberação de Medicamentos , Humanos , MicroRNAs/química , Conformação de Ácido Nucleico
15.
Neurochem Int ; 139: 104808, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32711020

RESUMO

The molecular mechanism responsible for hyperphosphorylated tau accumulation in dendrites of Alzheimer's disease (AD) neurons has not been fully clarified. Recently, we reported that tau mRNA is distributed into dendrites, and that translation and phosphorylation of tau protein are immediately enhanced in response to glutamatergic stimulation. Here, we focused on dendritic glycogen synthase kinase 3ß (GSK3ß), a key enzyme for tau phosphorylation, and investigated the mechanism responsible for the neural stimulation-induced hyperphosphorylation of the newly translated dendritic tau protein. We found that GSK3ß mRNA was also distributed into dendrites of cultured hippocampal neurons, and that a glutamate-dependent slight increase of translation occurred in a short time. Concomitantly, dephosphorylation at the Ser9 residue of the preexisting GSK3ß, which reactivates this kinase, was strongly induced without an increase of its phosphatase PP1 or a decrease of the PP1 inhibitor I-2. Instead, I-2 phosphorylation was observed, suggesting disinhibition of PP1. This glutamate-dependent phosphorylation of I-2 and the dephosphorylation of preexisting GSK3ß were abolished in the presence of GSK3ß inhibitors. Interestingly, translational obstruction of GSK3ß mRNA also canceled these reactions. These results indicate that dendrites exhibit a glutamate-responsive cycle for amplification of reactivated preexisting GSK3ß operating via PP1 disinhibition, whose activation requires neural activity-dependent translation of dendritic GSK3ß mRNA. This would explain why a slight increase of dendritic GSK3ß is sufficient to trigger hyperphosphorylation of significantly increased tau protein.


Assuntos
Dendritos/metabolismo , Ácido Glutâmico/toxicidade , Glicogênio Sintase Quinase 3 beta/metabolismo , Biossíntese de Proteínas/fisiologia , RNA Mensageiro/metabolismo , Animais , Dendritos/efeitos dos fármacos , Dendritos/genética , Glicogênio Sintase Quinase 3 beta/genética , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fosforilação/efeitos dos fármacos , Fosforilação/fisiologia , Biossíntese de Proteínas/efeitos dos fármacos , RNA Mensageiro/genética , Proteínas tau
16.
Biotechnol Prog ; 36(6): e3016, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32390308

RESUMO

Quality control of monoclonal antibodies is challenging due in part to the diversity of post-translational modifications present. The regulation of the N-glycans of IgG-Fc domain is one of the key factors to maintain the safety and efficacy of antibody drugs. The FcγRIIIa affinity column is an attractive tool for the precise analysis of the N-glycans in IgG-Fc domain. We used the mutant FcγRIIIa, which is produced in Escherichia coli and is therefore not glycosylated, as an affinity reagent to analyze the N-glycans of monoclonal antibodies expressed in Expi293 and ExpiCHO cells. The monoclonal antibodies expressed in these cells showed very different chromatograms, because of differences in terminal galactose residues on the IgG-Fc domains. We also carried out kinetic and thermodynamic analyses to understand the interaction between monoclonal antibodies and the mutant FcγRIIIa. Expi293 cell-derived monoclonal antibodies had higher affinity for the mutant FcγRIIIa than those derived from ExpiCHO cells, due to slower off rates and lower binding entropy loss. Collectively, our results suggest that the FcγRIIIa column can be used to analyze the glycosylation of antibodies rapidly and specifically.


Assuntos
Anticorpos Monoclonais/isolamento & purificação , Fragmentos Fc das Imunoglobulinas/isolamento & purificação , Imunoglobulina G/isolamento & purificação , Polissacarídeos/isolamento & purificação , Receptores de IgG/genética , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/imunologia , Células CHO , Cromatografia Líquida de Alta Pressão , Cricetinae , Cricetulus , Galactose/genética , Glicosilação , Células HEK293 , Humanos , Fragmentos Fc das Imunoglobulinas/química , Fragmentos Fc das Imunoglobulinas/genética , Fragmentos Fc das Imunoglobulinas/imunologia , Imunoglobulina G/química , Imunoglobulina G/genética , Imunoglobulina G/imunologia , Ligantes , Polissacarídeos/química , Polissacarídeos/genética , Processamento de Proteína Pós-Traducional/genética , Receptores de IgG/imunologia
17.
Int J Mol Sci ; 21(8)2020 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-32295066

RESUMO

Helicobacter pylori is associated with the onset of gastritis, peptic ulcers, and gastric cancer. Galectins are a family of ß-galactoside-binding proteins involved in diverse biological phenomena. Galectin-2 (Gal-2), a member of the galectin family, is predominantly expressed in the gastrointestinal tract. Although some galectin family proteins are involved in immunoreaction, the role of Gal-2 against H. pylori infection remains unclear. In this study, the effects of Gal-2 on H. pylori morphology and survival were examined. Gal-2 induced H. pylori aggregation depending on ß-galactoside and demonstrated a bactericidal effect. Immunohistochemical staining of the gastric tissue indicated that Gal-2 existed in the gastric mucus, as well as mucosa. These results suggested that Gal-2 plays a role in innate immunity against H. pylori infection in gastric mucus.


Assuntos
Galactosídeos/farmacologia , Galectina 2/farmacologia , Helicobacter pylori/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Animais , Infecções por Helicobacter , Helicobacter pylori/crescimento & desenvolvimento , Humanos , Masculino , Camundongos
18.
Biol Pharm Bull ; 43(4): 742-746, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32238717

RESUMO

Roundabout4 (Robo4) is an endothelial cell-specific protein that stabilizes the vasculature in pathological angiogenesis and inflammation. We previously determined a 3-kb Robo4 promoter and demonstrated the importance of the upstream region for nuclear factor-kappaB (NF-κB)-mediated promoter activation induced by tumor necrosis factor α (TNFα). This region contains unique genomic features, including promoter region-specific DNA hypermethylation and chromatin condensation; however, the function of the region remains poorly understood. In this study, we analyzed the DNA sequences of the region and identified a motif for polycomb repressive complex 2 (PRC2). Chromatin immunoprecipitation assay indicates the binding of the PRC2 component, SUZ12, to the motif. A mutation in the motif decreased DNA methylation in embryonic stem cells and increased Robo4 promoter activity in endothelial cells. An inhibitor for the PRC2 component, EZH2, induced the promoter activity and expression of Robo4 in endothelial cells treated with or without TNFα. Taken together, these results indicate that the PRC2 components maintain DNA hypermethylation and suppress Robo4 expression via the PRC2 binding motif in the upstream promoter.


Assuntos
Metilação de DNA , Células Endoteliais da Veia Umbilical Humana/metabolismo , Complexo Repressor Polycomb 2/metabolismo , Regiões Promotoras Genéticas , Receptores de Superfície Celular/genética , Animais , Células Cultivadas , Células-Tronco Embrionárias/metabolismo , Proteína Potenciadora do Homólogo 2 de Zeste/farmacologia , Regulação da Expressão Gênica , Humanos , Camundongos , Fator de Necrose Tumoral alfa/farmacologia
19.
Biol Pharm Bull ; 43(2): 356-360, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32009121

RESUMO

Galectins are a group of animal lectins characterized by their specificity for ß-galactosides. Of these, galectin-2 (Gal-2) is predominantly expressed in the gastrointestinal tract. In the current study, we used a mouse gastric mucous fraction to investigate whether Gal-2 is secreted from epithelial cells and identify its potential ligands in gastric mucus. Gal-2 was detected in the mouse gastric mucous fraction and could be eluted from it by the addition of lactose. Affinity chromatography using recombinant mouse galectin-2 (mGal-2)-immobilized adsorbent and subsequent LC-MS/MS identified MUC5AC, one of the major gastric mucin glycoproteins, as a potential ligand of mGal-2. Furthermore, MUC5AC was detected in the mouse gastric mucous fraction by Western blotting, and recombinant mGal-2 was adsorbed to this fraction in a carbohydrate-dependent manner. These results suggested that Gal-2 and MUC5AC in mouse gastric mucus interact in a ß-galactoside-dependent manner, resulting in a stronger barrier structure protecting the mucosal surface.


Assuntos
Galectina 2/química , Trato Gastrointestinal/química , Mucina-5AC/química , Animais , Humanos , Lactose , Camundongos , Muco , Estômago
20.
Thorac Cancer ; 11(4): 1052-1060, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32096610

RESUMO

BACKGROUND: Interstitial lung disease (ILD) induced by immune checkpoint inhibitors (ICIs) is a potentially life-threatening adverse event. The purpose of this study was to evaluate whether the development of immune-related adverse events (irAEs), especially ILD, was associated with treatment efficacy and to research the features and risk factors of ILD in advanced non-small cell lung cancer (NSCLC). METHODS: Between December 2015 and November 2018, 130 advanced NSCLC patients were treated with nivolumab, pembrolizumab or atezolizumab. The patients were categorized into two groups (irAEs group or non-irAEs group). Subsequently, we divided the irAEs group into two groups based on the incidence of ILD (ILD group and irAEs-non-ILD group). Treatment efficacy and the characteristics of ILD were evaluated. RESULTS: A total of 39 (30%) patients developed irAEs. ILD was observed in 16 (12%) patients. Patients with ILD had a higher objective response rate (ORR) compared with irAEs-non-ILD patients and non-irAEs patients (63%, 43% and 22%, respectively). Median progression-free survival (mPFS) was 15.9 months in ILD patients, 5.4 months in irAEs-non-ILD patients and 3.3 months in non-irAEs patients (log-rank test, P = 0.033). Pre-existing interstitial pneumonia (IP) was an independent risk factor for ILD-induced ICIs (odds ratio [OR] 14.7; 95% confidence interval [CI]: 2.16-99.6, P = 0.006). CONCLUSIONS: ORR and PFS were significantly better in ILD patients than in irAEs-non-ILD and non-irAEs patients. Pre-existing history of IP was an independent risk factor for ILD-induced ICIs.


Assuntos
Adenocarcinoma de Pulmão/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma de Células Escamosas/tratamento farmacológico , Inibidores de Checkpoint Imunológico/efeitos adversos , Doenças Pulmonares Intersticiais/mortalidade , Neoplasias Pulmonares/tratamento farmacológico , Adenocarcinoma de Pulmão/imunologia , Adenocarcinoma de Pulmão/patologia , Idoso , Carcinoma Pulmonar de Células não Pequenas/imunologia , Carcinoma Pulmonar de Células não Pequenas/patologia , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/patologia , Feminino , Seguimentos , Humanos , Doenças Pulmonares Intersticiais/induzido quimicamente , Doenças Pulmonares Intersticiais/patologia , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/patologia , Masculino , Prognóstico , Estudos Retrospectivos , Taxa de Sobrevida
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