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1.
Auris Nasus Larynx ; 2021 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-34728118

RESUMO

OBJECTIVE: Transglutaminase (TGM)2 and peroxisome proliferator-activated receptor (PPAR)γ are thought to participate in the pathogenesis of nasal polyp formation in cystic fibrosis (CF). We herein investigated expressions of cystic fibrosis transmembrane conductance regulator (CFTR), TGM2, PPARγ and isopeptide bonds, a reaction product of TGM, in non-CF nasal polyps. METHODS: Nasal polyps and inferior turbinates were collected from chronic rhinosinusitis patients without CF during transnasal endoscopic sinonasal surgery. Expressions of CFTR, TGM2, isopeptide bonds and PPARγ were examined by fluorescence immunohistochemistry and quantitative RT-PCR. Expression of CFTR was also analyzed by Western blot. RESULTS: Immunohistochemical fluorescence of the nasal polyp was significantly lower for CFTR and PPARγ, and significantly higher for TGM2 and isopeptide bonds than that of the turbinate mucosa. Lower expression of CFTR in the nasal polyp than in the turbinate mucosa was also observed in Western blot. Expression of PPARG mRNA was significantly lower in the nasal polyp than in the turbinate mucosa, whereas expressions of CFTR mRNA or TGM2 mRNA did not differ between the two tissues. Immunohistochemical fluorescence for CFTR showed significant negative correlation with that for TGM2 and isopeptide bonds, and significant positive correlation with that for PPARγ. The fluorescence for TGM2 was positively correlated with that for isopeptide bonds and negatively correlated with that for PPARγ. The fluorescence for isopeptide bonds tended to be negatively correlated with that for PPARγ. CONCLUSIONS: These results suggest a possible role of the CFTR-TGM2-PPARγ cascade in the pathogenesis of nasal polyp formation in non-CF patients as in CF patients.

2.
Dent Mater J ; 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34602588

RESUMO

This study aimed to evaluate the pulpal responses of monkey's pulp after direct pulp capping (DPC) with the novel mineral trioxide aggregate containing phosphorylated pullulan-based material (MTAPPL). Seventy-two teeth were randomly divided into four groups: MTAPPL; Nex-Cem MTA (NX); TheraCal LC (TH); and Dycal (DY). Histopathological changes in the pulps were observed at days 3, 7 and 70. On day 3, mild inflammatory responses were observed in the MTAPPL, no to moderate inflammatory responses in the TH, whereas moderate inflammatory responses in the NX and DY. No mineralized tissue formation (MTF) was observed in all groups. On day 7, no or mild inflammatory responses were observed in all groups. Initial MTF was observed except for DY. No inflammation with complete MTF including presence of odontoblast-like cells was observed in the MTAPPL, NX and TH groups at day 70. These findings indicate that MTAPPL could be an efficient DPC material.

3.
Materials (Basel) ; 14(19)2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34640141

RESUMO

This study prepared glass ionomer cement (GIC) containing nanoporous silica (NPS) (GIC-NPS) at 5 wt% concentrations using 3 types of NPS with different pore and particle sizes and evaluated the differences in their cationic ion capture/release abilities and mechanical properties. The cationic water-soluble dye was used as cationic ion. The test GIC-NPS complexes captured dyes by immersion in 1 wt% dye solutions. All the GIC-NPS complexes released dyes for 28 d, and the amount of dye released from the complexes increased with decreasing pore size; however, the particle size of NPS did not affect the amount of dye released. Additionally, GIC-NPS was able to recharge the dye, and the amount of released the dye by the complexes after recharge was almost identical to the amount released on the first charge. Although not significantly different, the compressive strength of GIC-NPS was slightly greater than that of GIC without NPS regardless of the type of NPS. These results suggest that the degree of capture and release of cationic molecules, such as drugs, can be controlled by optimizing the pore size of NPS without sacrificing its mechanical strength when its content is 5 wt%.

4.
BMC Nephrol ; 22(1): 295, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-34461850

RESUMO

BACKGROUND: In typical cases of Bartter syndrome (BS), assessing response to diuretics (furosemide and thiazide), hereinafter referred to as diuretic loading test, may be used to diagnose the type by detecting which part of the kidney tubule is not functioning correctly. However, the diuretic loading test may not always agree with the results of genetic analyses. CASE PRESENTATION: A 5-year-old boy was admitted due to lower extremity weakness and abnormal gait. He had a recurrent episode of muscle weakness and laboratory results showed severe hypokalemia. The direct genomic sequencing of the case revealed a new mutation in the SLC12A1 gene, which is associated with type I Bartter syndrome. Because there was the difference between the phenotype and genotype, we conducted a diuretic loading test to confirm the diagnosis. However, the results showed a clear increase in urine excretion of Na and Cl. These results were not consistent with typical type I BS, but consistent with the patient's phenotype. CONCLUSION: The diuretic loading test has limited utility for diagnosis especially in atypical cases. On the other hand, this test, which allows assessment of channel function, is useful for better understanding of the genotype-phenotype correlation.

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.
Dev Comp Immunol ; 122: 104124, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-33974965

RESUMO

DNA damage-induced cellular senescence is involved in aging. We reported previously that p53+/- mice subjected to irradiation at a young age exhibited an increased number of splenic lymphocytes in the S and G2/M phases. However, the detailed nature of splenic disorders in these mice is not fully understood. In this study, we investigated the effects on molecules in splenocytes, especially on senescence factors after early exposure of mice to radiation. Mice, 8- (young) or 17-, 30-, and 41-week-old (old) p53+/- were subjected to 3-Gy whole-body irradiation. Splenocytes were prepared at 56 weeks of age. Immunoblot showed that irradiation at 8 weeks enhanced the expression and phosphorylation of p53, cyclin-dependent kinase 2, cell division cycle 6, and the MDM2 proto-oncogene in splenocytes. However, these molecules were not affected by irradiation at 17, 30, and 41 weeks of age. Similarly, irradiation at 8, but not 17, 30, or 41 weeks, induced phosphorylation of IKKα, NF-κB inhibitor alpha, and p65. Electrophoretic mobility shift assay demonstrated that active forms of NF-κB were increased. In addition, enzyme-linked immunosorbent assay showed that lipopolysaccharide-induced IL-6 production was enhanced in splenocytes of mice irradiated at 8 weeks. ATP levels were increased in splenocytes of mice irradiated at 8, but not 17, 30, or 41 weeks. CDK2 expression and p65 phosphorylation were induced in CD45R/B220+ cells from irradiated mice. Overall, irradiation induced a NF-κB-related immune response in the spleen with an increase in senescence marker proteins, such as CDKs and IL-6, which are known to be typical senescence-associated secretory phenotype factors related to stresses, such as DNA damage.

7.
Materials (Basel) ; 14(6)2021 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-33809621

RESUMO

Several dental materials contain silver for antibacterial effect, however the effect is relatively low. The reason for the lower antibacterial efficacy of silver is considered to be the fact that silver ions bind to chloride ions in saliva. To develop new effective silver antibacterial agents that can be useful in the mouth, we synthesized two novel amino acid (methionine or histidine)-silver complexes (Met or His-Ag) loaded with montmorillonite (Mont) and analyzed their antibacterial efficacy. At first the complexes were characterized using nuclear magnetic resonance (NMR), and amino acid-Ag complex-loaded Mont (amino acid-Ag-Mont) were characterized using X-ray diffraction (XRD) and scanning electron microscopy (SEM). The antibacterial efficacy of these materials in dental acrylic resin was then investigated by bacterial growth measurement using a spectrophotometer. As controls, commercially available silver-loaded zeolite and silver-zirconium phosphate were also tested. Dental acrylic resin incorporating His-Ag-Mont strongly inhibited Streptococcus mutans growth. This was explained by the fact that His-Ag complex revealed the highest amounts of silver ions in the presence of chloride. The structure of the amino acid-Ag complexes affected the silver ion presence in chloride and the antibacterial efficacy. His-Ag-Mont might be used as antibacterial agents for dental materials.

8.
Dent Mater ; 37(6): e391-e398, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33757654

RESUMO

OBJECTIVE: Bisphenol A-glycidyl methacrylate (Bis-GMA) and urethane dimethacrylate (UDMA) are widely used as the primary components of (meth)acrylic monomers. However, the use of Bis-GMA, which is a bisphenol A derivative, in dentistry is being questioned after bisphenol A was found to exhibit estrogenic activity. Although UDMA is being considered as a substitute for bis-GMA, the mechanical properties of cured resin composites containing UDMA are less than desirable. Therefore, in this study, we developed new alternative (meth)acrylic monomers to enhance the mechanical strength of cured composite resins. METHODS: Five urethane acrylic monomers were synthesized in this study as (meth)acrylic monomer substituents to replace Bis-GMA and UDMA. The elastic modulus, strength, and breaking energy values of cured resins consisting of mixtures of the urethane acrylates and diluting monomers were determined using the three-point flexural test. The data obtained were analyzed using one-way ANOVA and the post-hoc Tukey HSD tests (p < 0.05). Viscosities of the urethane acrylic monomers were measured with a cone-plate viscometer. Refractive indices of the urethane acrylic monomers were determined with an Abbe refractometer. RESULTS: The results of the three-point flexural tests revealed that the cured urethane acrylic monomer-based resin exhibited higher elastic modulus (up to 40%) and strength (up to 21%) compared to the cured UDMA-based resin. Viscosity and refractive index of the urethane acrylic monomers were between those of UDMA and Bis-GMA. SIGNIFICANCE: The developed urethane diacrylates prepared from diisocyanates which have an aromatic or aliphatic ring, 1,3-bis(1-isocyanato-1-methylethyl)benzene (TMXDI), 1,3-bis(isocyanatomethyl)benzene (XDI), or norbornane diisocyanate (NBDI) are worth considering as alternative options of Bis-GMA and UDMA for restorative resin composites.


Assuntos
Metacrilatos , Poliuretanos , Bis-Fenol A-Glicidil Metacrilato , Resinas Compostas , Teste de Materiais , Polietilenoglicóis , Ácidos Polimetacrílicos
9.
Int J Mol Sci ; 22(5)2021 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-33652591

RESUMO

Repressor element-1 (RE-1) or neural restrictive silencer element (NRSE) bound with a zinc finger transcription repressor, RE-1 silencing transcription factor (REST, also known as neural restrictive silencer factor, NRSF) has been identified as a fundamental repressor element in many genes, including neuronal genes. Genes regulated by REST/NRSF regulate multifaceted neuronal phenotypes, and their defects in the machinery cause neuropathies, disorders of neuron activity), autism and so on. In REST repressions, the N-terminal repressor domain recruits Sin3B via its paired amphipathic helix 1 (PAH1) domain, which plays an important role as a scaffold for histone deacetylase 1 and 2. This machinery has a critical role in maintaining neuronal robustness. In this study, in order to establish protein-protein interaction assays mimicking a binding surface between Sin3B and REST, we selected important amino acids from structural information of the PAH1/REST complex and then tried to reconstitute it using recombinant short peptides derived from PAH1/REST. Initially, we validated whether biotinylated REST interacts with glutathione S-transferase (GST)-tagged PAH1 and whether another PAH1 peptide (PAH1-FLAG) competitively binds with biotinylated REST using surface plasmon resonance (SPR). We observed a direct interaction and competitive binding of two PAH1 peptides. Secondly, in order to establish a high-throughput and high-dynamic-range assay, we utilized an easily performed novel time-resolved fluorescence energy transfer (TR-FRET) assay, and closely monitored this interaction. Finally, we succeeded in establishing a novel high-quality TR-FRET assay and a novel interaction assay based on SPR.


Assuntos
Transferência Ressonante de Energia de Fluorescência , Proteínas Repressoras/química , Ressonância de Plasmônio de Superfície , Humanos , Ligação Proteica , Proteínas Repressoras/metabolismo
10.
Invest New Drugs ; 39(1): 122-130, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-32914311

RESUMO

Adult T cell leukemia (ATL) is an aggressive and malignant blood disease. We previously reported that steroid-structured cucurbitacin D (CuD) induces apoptosis in ATL cells. In this study, we investigated the effects of mitogen-activated protein kinase (MAPK) signaling inhibitors on CuD-induced cell death in peripheral blood lymphocytes (PBLs) isolated from ATL/acute lymphoblastic leukemia (ALL) patients and two human leukemia cell lines (MT-1 and MT-4). PBLs were isolated from an ATL/ALL patient as well as from a healthy donor. Cell surface markers were examined using flow cytometry. Serum cytokine levels were estimated using LEGENDplex or analyzed at the Center for Clinical and Translational Research of Kyushu University Hospital. Cell proliferation was assessed using the Cell Titer-Glo luminescent cell viability assay. Protein expression was determined by western blotting. PBLs from patients highly expressed CD4 and CD5. Serum from the patient contained high levels of interleukin (IL)-8, IL-10, IL-18, and interferon-γ compared to the healthy donor. CuD-induced cell death was enhanced by the mitogen-activated protein kinase kinase (MEK)1/2 inhibitor U0126. However, a c-Jun N-terminal kinase (JNK) inhibitor prevented CuD-induced cell death. Immunoblot analyses revealed that CuD reduced the phosphorylation of extracellular signal-regulated kinase (ERK), p38, and JNK, and co-treatment with CuD and U0126 did not affect the phosphorylation of ERK. MEK1/2 and p38 inhibitors enhanced CuD-induced cell death, and U0126 enhanced the CuD-induced de-phosphorylation of ERK in MT-1 and MT-4 cells. We conclude that CuD reduces ERK activation, resulting in enhanced antitumor effects on leukemic cells.

11.
J Thorac Dis ; 12(11): 6523-6532, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33282354

RESUMO

Background: Early stage non-small cell lung cancer (NSCLC) is good candidate for video-assisted thoracoscopic surgery (VATS). Long-term outcome compared between VATS and open surgery remains unclear. The aim of this study was to assess the long-term outcome of VATS in early stage adenocarcinoma. Methods: A retrospective study was performed in 546 patients which were operated between January 2006 and December 2010 in our institute and of those, 240 (220 lobectomies, and 20 segmentectomies) were clinical N0 adenocarcinoma. One hundred and thirty-five patients underwent VATS and 105 patients for open surgery. Long-term oncological outcomes were compared in both groups. Results: There were significant differences in age, gender, Blinkman index, clinical T factor and tumor size between two groups. VATS group showed statistically longer operation time (P=0.01), less blood loss (P=0.005), shorter length of stay (P=0.001), and less dissected number of lymph nodes (P<0.001) compared with open surgery. Disease-free survival in VATS was significantly better than open surgery (5- and 10-year survival; VATS, 91.4%, 79.0%; open, 85.1%, 73.6%; respectively, P=0.04). Overall survival in VATS was not different from open (P=0.58). Propensity matched disease-free and overall survival was not significantly different between two groups. Multivariate Cox regression analysis showed that age [P=0.04, 95% confidence interval (CI): (1.02-6.81)] in overall and T factor [P=0.01, 95% CI: (1.41-17.3)] in disease-free survival was prognostic significant after propensity matching. Conclusions: Our study demonstrated that long-term outcome in VATS for early stage adenocarcinoma was equivalent to open surgery. VATS may be a treatment of choice for promising long-term prognosis.

12.
J Toxicol Environ Health A ; 83(19-20): 649-658, 2020 10 17.
Artigo em Inglês | MEDLINE | ID: mdl-32819208

RESUMO

Air pollution, especially that initiated by particulate matter (PM), has been implicated as a risk factor for several inflammatory diseases. Previously, it was reported that PM enhances immune responses. PM includes the tar fraction that contains polycyclic aromatic hydrocarbons (PAHs), which produce adverse health effects in exposed individuals. However, the influence of the tar fraction (as a component of PM) on splenocytes is not fully understood. The aim of this study was to determine the effects of the tar fraction extracted from PM collected from the atmosphere in Fukuoka, Japan, on mouse splenocytes. ICR mice were administered tar (1 or 5 µg/mouse) intratracheally 4 times at 2-week intervals, and splenocytes from the tar-treated mice were extracted and examined. The parameters determined were proliferation, cytokine concentrations and transcription factors activation. Following tar treatment, splenocyte proliferation increased relative to controls. Concanavalin A (ConA)-induced interleukin (IL)-2 formation and ConA- or lipopolysaccharide (LPS)-induced interferon-γ production were elevated in splenocytes from tar-exposed mice. However, the production of tumor necrosis factor-α and IL-6 induced by LPS was not markedly changed following tar treatment. Further, nuclear factor of activated T cells, but not nuclear factor-κB, was enhanced in splenocytes of tar-exposed mice. Data indicate that tar-activated splenocytes and PM-bound PAHs might contribute to T cell activation in the spleen.


Assuntos
Poluentes Atmosféricos/imunologia , Poeira/imunologia , Material Particulado/imunologia , Hidrocarbonetos Policíclicos Aromáticos/imunologia , Baço/efeitos dos fármacos , Poluentes Atmosféricos/efeitos adversos , Poluição do Ar/efeitos adversos , Animais , Japão , Masculino , Camundongos , Camundongos Endogâmicos ICR , Material Particulado/efeitos adversos , Hidrocarbonetos Policíclicos Aromáticos/administração & dosagem , Areia
13.
J Occup Health ; 62(1): e12135, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32715571

RESUMO

OBJECTIVES: Although 1-Bromopropane (1-BP) exposure has been reported to cause neurotoxicity in adult humans and animals, its effects on the development of the central nervous system remain unclear. Recently, we reported delayed developmental neurotoxicity (DNT) upon 1-BP exposure in rats. Here we aimed to study the effect of prenatal 1-BP exposure on the hippocampal excitability in the juvenile offspring. METHODS: Pregnant Wistar rats were exposed to vaporized 1-BP for 20 days (6 h/d) with concentrations of 0 (control), 400, or 700 ppm. Hippocampal slices were prepared from male offspring during postnatal days (PNDs) 13, 14, and 15. Field excitatory postsynaptic potential (fEPSP) and population spike (PS) were recorded simultaneously from the CA1 region. RESULTS: In the exposed groups, the stimulation/response relationships of fEPSP slope and PS amplitude were enhanced more than in the control group at PND 14. Analysis of fEPSP-spike coupling demonstrated increased values of Top and Eslope50 in the exposed groups. Real-time PCR analysis showed a significant increase in the mRNA levels of the adult type Nav 1.1 Na+ channel subunit and the GluR1 glutamate receptor subunit in the hippocampus of the 700 ppm group at PND 14. CONCLUSIONS: Our results provide evidence that prenatal exposure to 1-BP accelerates developmental enhancement of hippocampal excitability in the pups before eye-opening. The current study suggests that our evaluation method of DNT is applicable to the industrial chemical 1-BP.


Assuntos
Região CA1 Hipocampal/efeitos dos fármacos , Excitabilidade Cortical/efeitos dos fármacos , Exposição por Inalação , Lactação , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Animais , Feminino , Hidrocarbonetos Bromados/efeitos adversos , Gravidez , Ratos , Ratos Wistar
14.
Dent Mater ; 36(7): 914-926, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32473833

RESUMO

OBJECTIVE: Hydrofluoric-acid etching followed by silanization is a routine clinical protocol for durable bonding to glass ceramics. Simplifying ceramic-bonding procedures, new technological developments involve the inclusion of a silane coupling agent in a self-adhesive composite cement. To investigate the effectiveness of the incorporated silane coupling agent, shear bond strength (SB) to ceramic and dentin, contact angle of water (CA), transmission electron microscopy (TEM), X-ray diffraction (XRD) and 29Si nuclear magnetic resonance (NMR) assessments were correlatively conducted. MATERIALS AND METHODS: SB to glass ceramic was measured without ('immediate') and with ('aged') 50K thermocycles upon application of (1) the silane-containing self-adhesive composite cement Panavia SA Cement Universal ('SAU'), being light-cured: 'SAU_light', (2) 'SAU_chem': chemically cured SAU, (3) 'SAP_light': light-cured Panavia SA Cement Plus ('SAP'), and (4) 'SAP_CP': SAP light-cured after separate silanization using Clearfil Ceramic Primer Plus ('CP'). CA was also measured on glass ceramic. The cement pastes before and upon mixing were characterized using 29Si NMR. SB of SAU or SAP onto dentin was measured. Finally, the cement-dentin interface was characterized by TEM and XRD. RESULTS: The immediate and aged SB to glass ceramic of SAU did not significantly differ from those of SAP_CP, while they were significantly higher than those of SAP. CA of SAU did not significantly differ from that of SAP_CP, but it was significantly higher than CA of SAP. 29Si NMR revealed siloxane bonds after mixture. SB of SAU and SAP to dentin did not show any significant difference. SEM, TEM and XRD confirmed tight and chemical interaction, respectively. SIGNIFICANCE: Incorporating silane in a 10-MDP-based self-adhesive composite cement combined efficient silane-coupling ability at the ceramic surface with effective bonding ability at dentin.


Assuntos
Colagem Dentária , Cimentos de Resina , Cerâmica , Porcelana Dentária , Teste de Materiais , Silanos , Propriedades de Superfície
15.
Int Immunopharmacol ; 84: 106510, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32361568

RESUMO

Reports show that particulate matter (PM) is related to respiratory and cardiovascular diseases. We previously reported the biological effects of PM in vivo and the endocytosis of PM by primary neutrophils from mice. Cell lines can be used to elucidate the mechanism underlying immune responses in detail; however, information is limited regarding the functions of neutrophils after PM exposure. Here, we investigated the immune response of primary neutrophils and dimethyl sulfoxide (DMSO)- and all-trans retinoic acid (ATRA)-differentiated HL-60 (neutrophil-like) cells to PM. We showed that endocytosis by ATRA-HL cells was enhanced compared to that by DMSO-HL cells and that endocytosis in both cells was inhibited by dynamin inhibitors. A MEK inhibitor, but not p38 or JNK inhibitors, inhibited endocytosis. The MEK inhibitor also inhibited the differentiation of ATRA-HL cells to neutrophils. We identified that endocytosis of PM by neutrophils activated the MAPK ERK and p38 pathways. DMSO-HL and ATRA-HL cells both produced TNF-α and IL-8 after lipopolysaccharide (LPS) or PM treatment, whereas non-differentiated HL-60 cells did not. MCP-1 production was enhanced in DMSO-HL cells after LPS or PM treatment, whereas it was high in ATRA-HL cells. Reactive oxygen species (ROS) production was enhanced after PM treatment to DMSO-HL cells. Further, extracellular extracts promoted endocytosis. The MEK inhibitor also reduced the production of TNF-α, IL-8, and MCP-1. Taken together, ERK activation is key for both differentiation and endocytosis, and DMSO-HL cells at day 6 can serve as a model of inflammatory neutrophils, such as bronchus neutrophils, and a good tool to analyze the molecular events involved in immune responses to PM.


Assuntos
Proteínas Quinases Ativadas por Mitógeno/imunologia , Neutrófilos/imunologia , Animais , Diferenciação Celular/efeitos dos fármacos , Citocinas/imunologia , Dimetil Sulfóxido/farmacologia , Endocitose/efeitos dos fármacos , Células HL-60 , Humanos , Inflamação/imunologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Neutrófilos/efeitos dos fármacos , Material Particulado/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Tretinoína/farmacologia
16.
J Appl Toxicol ; 40(9): 1239-1247, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32368826

RESUMO

We previously demonstrated that particulate matter ≤2.5 µm (PM2.5) suppresses the immune response in the spleen in vivo. Although PM2.5 includes the polycyclic aromatic hydrocarbon (PAH) such as dibenzo[a,h]anthracene (DBA), it is unclear whether PAH has a direct effect on the responses of splenocytes. In our study, the concentration of DBA used was approximately 0.8 µm, which is much lower than concentrations used in other toxicological studies of DBA. Although exposure to high concentrations of DBA is implicated in carcinogenesis, the effects of low doses of DBA on immune cells in vivo remain unclear. Here, we investigated the effects of low DBA doses on mouse splenocytes in vivo. Mice were administered dimethyl sulfoxide or DBA (0.4 or 0.8 µm) intratracheally. Twenty-four hours after treatment, the mice were killed and their splenocytes were collected. DBA treatment enhanced mitogen-induced cell proliferation and cytokine production in the mouse splenocytes. Furthermore, DBA enhanced splenic CD4+ and CD8+ cell proliferation and cytokine production. The nuclear factor of activated T cells (NFAT) was activated in CD4+ cells. DBA also activated nuclear factor-kappa B and CCAAT enhancer-binding protein pathways in CD11b+ cells. DBA-enhanced splenocyte activation was Toll-like receptor 2-, 4-, 9- and MyD88-independent. These results suggest that NFAT represents a promising marker for evaluation of the effects of DBA on T cells and T-cell-dependent antibody responses.

17.
J Mater Chem B ; 8(24): 5320-5329, 2020 06 24.
Artigo em Inglês | MEDLINE | ID: mdl-32458954

RESUMO

Several studies have shown the clinical success of hydraulic calcium-silicate cements (hCSCs) for direct and indirect pulp capping and root repair. However, hCSCs have various drawbacks, including long setting time, poor mechanical properties, low bond strength to dentin, and relatively poor handling characteristics. To overcome these limitations, a light-curable, resin-based hCSC (Theracal LC, Bisco) was commercially introduced; however, it did not exhibit much improvement in bond strength. We developed a light-curable self-adhesive pulp-capping material that contains the novel acrylamide monomer N,N'-{[(2-acrylamido-2-[(3-acrylamidopropoxy)methyl]propane-1,3-diyl)bis(oxy)]bis(propane-1,3-diyl)}diacrylamide (FAM-401) and the functional monomer 4-methacryloxyethyl trimellitate anhydride (4-MET). Two experimental resin-based hCSCs containing different calcium sources (portlandite: Exp_Pl; tricalcium silicate cement: Exp_TCS) were prepared, and the commercial hCSCs Theracal LC and resin-free hCSC Biodentine served as controls. The performance of each cement was evaluated based on parameters relevant for vital pulp therapy, such as curing degree on a wet surface, mechanical strength, as determined using a three-point bending test, shear bond strength to dentin, cytotoxicity, as determined using an MTT assay, and the amount of calcium released, as determined using inductively coupled plasma atomic emission spectrometry. Both experimental cements cured on wet surfaces and showed relatively low cytotoxicity. Furthermore, their flexural and shear bond strength to dentin were significantly higher than those of the commercial references. High calcium release was observed for both Exp_Pl and Biodentine. Thus, Exp_Pl as a new self-adhesive pulp-capping agent performed better than the commercial resin-based pulp-capping agent in terms of mechanical strength, bond strength, and calcium release.


Assuntos
Acrilamida/química , Agentes de Capeamento da Polpa Dentária e Pulpectomia/síntese química , Cimentos de Resina/síntese química , Acrilamida/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cricetinae , Cricetulus , Interações Hidrofóbicas e Hidrofílicas , Teste de Materiais , Estrutura Molecular , Tamanho da Partícula , Polimerização , Agentes de Capeamento da Polpa Dentária e Pulpectomia/química , Agentes de Capeamento da Polpa Dentária e Pulpectomia/farmacologia , Cimentos de Resina/química , Cimentos de Resina/farmacologia , Propriedades de Superfície
18.
Mar Drugs ; 18(3)2020 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-32183121

RESUMO

Toxoplasma gondii is a major protozoan parasite and infects human and many other warm-blooded animals. The infection leads to Toxoplasmosis, a serious issue in AIDS patients, organ transplant recipients and pregnant women. Neospora caninum, another type of protozoa, is closely related to Toxoplasma gondii. Infections of the protozoa in animals also causes serious diseases such as Encephalomyelitis and Myositis-Polyradiculitis in dogs or abortion in cows. Both Toxoplasma gondii and Neospora caninum have similar nucleoside triphosphate hydrolases (NTPase), NcNTPase and TgNTPase-I in Neospora caninum and Toxoplasma gondii, respectively. These possibly play important roles in propagation and survival. Thus, we targeted the enzymes for drug discovery and tried to establish a novel high-standard assay by a combination of original biochemical enzyme assay and fluorescent assay to determine ADP content. We then validated whether or not it can be applied to high-throughput screening (HTS). Then, it fulfilled criterion to carry out HTS in both of the enzymes. In order to identify small molecules having inhibitory effects on the protozoan enzyme, we also performed HTS using two synthetic compound libraries and an extract library derived from marine bacteria and then, identified 19 compounds and 6 extracts. Nagasaki University collected many extracts from over 18,000 marine bacteria found in local Omura bay, and continues to compile an extensive collection of synthetic compounds from numerous drug libraries established by Japanese chemists.


Assuntos
Medições Luminescentes , Neospora/enzimologia , Nucleosídeo-Trifosfatase/análise , Toxoplasma/enzimologia , Animais , Humanos
19.
Sci Rep ; 10(1): 4387, 2020 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-32152430

RESUMO

The remineralization mechanism in dental caries lesions is not completely understood. This study reports on ultrastructural and chemical changes observed within arrested caries lesions. Carious human teeth were observed using scanning electron microscopy (SEM) and focused-ion-beam (FIB)-SEM. The crystals detected in the caries lesions were characterized by transmission electron microscopy (TEM), along with chemical element mapping using energy-dispersive spectroscopy (EDS)-STEM. FIB-SEM 3D reconstructions revealed a severely damaged dentin surface abundantly covered by bacteria. Although the dentin tubules were clogged up to a depth of 100 µm, bacterial invasion into dentin tubules was not observed. TEM crystal analysis and EDS-STEM revealed the presence of Ca and P, as well as of Mg within the HAp crystals deposited inside the dentin tubules. It was concluded that extensive remineralization with deposition of Mg-HAp crystals had occurred in dentin tubules of caries-arrested dentin. Understanding the natural remineralization process is thought to be helpful for developing clinical biomimetic remineralization protocols.


Assuntos
Cárie Dentária/diagnóstico por imagem , Cárie Dentária/patologia , Remineralização Dentária , Humanos , Imageamento Tridimensional , Microscopia Eletrônica de Varredura
20.
Dent Mater ; 36(4): 512-526, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32061443

RESUMO

OBJECTIVE: To evaluate, by means of an ex-vivo human tooth-culture model and in-vivo minipig animal study, the pulpal inflammatory reaction and reparative dentin-formation capacity of an injectable phosphopullulan-based calcium-silicate cement (GC, Tokyo, Japan) upon pulp capping, this in comparison with the commercial reference material Biodentine (Septodont). METHODS: For the ex-vivo tooth model, 9 freshly-extracted teeth from 3 different patients were pulp-capped with the experimental biomaterial (n = 3), Biodentine (n = 3) or left uncapped (control; n = 3). The teeth were kept in fresh culture medium for 4 weeks and, upon fixation three-dimensional Micro-CT and histology were performed. For the in-vivo animal study, 40 teeth from 3 minipigs were exposed and pulp capped with the experimental biomaterial containing phosphopullulan (n = 24) or Biodentine (n = 16) for 7 or 70 days. The inflammatory reaction and the tissue-regenerative potential was qualitatively and semi-quantitatively characterized using three-dimensional micro-CT and histology. RESULTS: Ex vivo, the treatment with the experimental phosphopullulan-based calcium-silicate cement and Biodentine stimulated the formation of fibrous tissue and mineralized foci. In vivo, early inflammatory reaction and regeneration of the pulp-tissue interface was promoted by both bioceramic materials after 7 and 70 days, respectively. SIGNIFICANCE: Our findings bring new insights into calcium-silicate-mediated dental pulp repair and regeneration. The novel ready-to-use and self-adhering functionalized calcium-silicate cement revealed effective pulpal repair potential.


Assuntos
Agentes de Capeamento da Polpa Dentária e Pulpectomia , Cimento de Silicato , Animais , Cálcio , Compostos de Cálcio , Polpa Dentária , Capeamento da Polpa Dentária , Combinação de Medicamentos , Humanos , Óxidos , Silicatos , Engenharia Tecidual
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