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To ensure the safe transportation and efficient utilisation of lignite, it is important to inhibit its spontaneous combustion. In this study, Shengli lignite (SL+) was used as the research object and ionic liquids (ILs) were used to pretreat the lignite to investigate their effect on the combustion performance of lignite. On this basis, the relationship between the structure and combustion performance of lignite with different structures (heat treatment, oxidation) after ILs treatment was investigated. Results indicated that the combustion of lignite treated with ILs shifted towards higher temperatures. The most pronounced effect was observed in coal samples treated with [BMIM]Cl (1-butyl-3-methylimidazolium chloride), with the maximum combustion rate corresponding to a temperature increase of approximately 57 °C compared to that of the untreated lignite. For the heat-treated lignite, the temperature corresponding to the maximum combustion rate was approximately 38 °C higher than that of the untreated lignite. After [BMIM]Cl treatment, the combustion performance of the heat-treated lignite changed very slightly. In contrast, for oxidised lignite, the temperature corresponding to the maximum combustion rate decreased by approximately 54 °C compared with that of the untreated lignite and increased by approximately 135 °C after treatment with [BMIM]Cl. The characterisation results show that the content of aliphatic hydrogen and oxygen-containing functional groups decreased in the heat-treated lignite, while the content of hydroxyl and carboxyl groups increased in the oxidised lignite. The microstructure of the heat-treated lignite after [BMIM]Cl treatment changed slightly. In contrast, in the oxidised lignite after [BMIM]Cl treatment, the content of hydroxyl and carboxyl groups decreased, whereas the content of ether (C-O-) structures increased. The increased content of ether (C-O-) structures improved the stability of the coal samples. It is believed that the inhibition of lignite combustion is mainly attributed to the high stability of the ether (C-O-) structures. The kinetic analysis demonstrated that the ILs treatment increased the activation energy of lignite combustion.
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In this study, 11 core coal samples were collected from deep-buried coalbed methane (CBM) reservoirs with burial depth intervals of 900-1500 m for gas estimation content by a direct method. In desorption experiments, the cumulative gas desorption data were recorded within 2 h in the field on the basis of the China National Standard method. For accuracy, two improved methods were proposed. The results show that the gas contents of deep-buried coal samples based on the China National Standard and mud methods are 3.58-9.89 m3/t (average of 6.03 m3/t) and 3.74-10.05 m3/t (average of 6.20 m3/t), respectively. The proposed Langmuir equation and logarithmic equation methods exhibited nonlinear relationships between the cumulative desorption volume and desorption time, which yield values of 6.33-13.34 m3/t (average of 9.36 m3/t) and 6.15-13.86 m3/t (average of 10.37 m3/t), respectively. In addition, the two proposed methods combine the raw data within 2 h by the China National Standard method and additional desorption points during extra time, which are helpful for the ability of the hypothetical methods to calculate the gas content. The Langmuir equation method is a relatively more accurate method to estimate the gas content in comparison with the proposed logarithmic method, which is based on the relative error and comparison plots of actual data and simulated results. From the perspective of numerical value, the Langmuir equation method gives values 1.06-3.39 times (average of 1.86 times) those of the China National Standard method. These analyses show that the proposed Langmuir equation method with extra desorption points is an effective method to determine the gas content of deep-buried CBM reservoirs.
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Natural processes and anthropogenic activities simultaneously control the long-term spatial and temporal variations of groundwater hydrogeochemistry in coalfields. In this study, the spatiotemporal variations and primary controlling factors of deep groundwater hydrogeochemistry in the Carboniferous limestone aquifer of the Huaibei coalfield, North China were investigated using cluster analysis combined with geological conditions, water-rock interactions and mining activities. The analysis data of 176 groundwater samples collected over five years from 20 monitoring wells were subdivided into six clusters through hierarchical cluster analysis. Moreover, principal component analysis, box plots and Piper and Stiff diagrams were employed to analyze the statistical and hydrogeochemical characteristics of each cluster, and to reveal the differences and connections between the clusters. The results show that there are significantly spatial variations in groundwater hydrogeochemistry, while the temporal variations are not evident with only a few notable exceptions. Geological conditions dominate the groundwater hydrogeochemistry by controlling the hydraulic connection between groundwater and meteoric water and the flow conditions of groundwater. Moreover, the types and degrees of diverse water-rock interactions in different regions are another important factor controlling the spatial variations of groundwater hydrogeochemistry. Anthropogenic activities are mainly pumping and drainage, which has led to the overall decline in groundwater levels and the temporal variations of hydrogeochemistry in some zones. The findings of this study not only have important implications for deep groundwater resources management in the Huaibei coalfield, but also provide a research template for other highly exploited coalfields in North China.
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Minas de Carbón , Agua Subterránea , Contaminantes Químicos del Agua , China , Monitoreo del Ambiente/métodos , Contaminantes Químicos del Agua/análisisRESUMEN
Both natural processes and anthropogenic activities have significant effects on groundwater evolution in coal mining regions. In this study, the primary controlling mechanism of the groundwater chemistry evolution for the Carboniferous groundwater in the Huaibei coalfield, North China was proposed based on the hydrogeochemical indicators combining with multiple isotope tracers. The diversity of hydrochemical types indicates the complexity of the hydrogeochemical environment in the groundwater, which is recharged by precipitation infiltration with minimal evaporation according to the distributions of δD and δ18O. Additionally, ion correlation analysis suggests that minerals dissolution and cation exchange between Na+ and Ca2+ are the dominant processes within that groundwater. The hydrochemical and δ13CDIC characteristics of the groundwater demonstrate that HCO3- is mainly controlled by the dissolution of carbonate minerals and soil CO2, and the proportion of the latter is believed to be dominated by the hydrogeologic conditions. Similarly, the values of SO42- and δ34SSO4 indicate that a small portion of SO42- in the groundwater in the northern part originates from the meteoric precipitation, while it is mainly derived from the dissolution of gypsum in the southern part. Furthermore, mining activities also alter the groundwater level and flow conditions through pumping and drainage, which enhances the interaction between groundwater and aquifer lithologies, thereby affects the hydrogeochemical processes. The findings of this work are of great significance for promoting the safe exploitation of deep coal resources and the sustainable utilization of groundwater in the Huaibei coalfield, as well as the most of other coalfields in North China.
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Minas de Carbón , Agua Subterránea , Contaminantes Químicos del Agua , China , Monitoreo del Ambiente , Isótopos/análisis , Contaminantes Químicos del Agua/análisisRESUMEN
The calcium release-activated calcium channel Orai regulates Ca2+ entry into non-excitable cells and is required for proper immune function. While the channel typically opens following Ca2+ release from the endoplasmic reticulum, certain pathologic mutations render the channel constitutively open. Previously, using one such mutation (H206A), we obtained low (6.7 Å) resolution X-ray structural information on Drosophila melanogaster Orai in an open conformation (Hou et al., 2018). Here we present a structure of this open conformation at 3.3 Å resolution using fiducial-assisted cryo-electron microscopy. The improved structure reveals the conformations of amino acids in the open pore, which dilates by outward movements of subunits. A ring of phenylalanine residues repositions to expose previously shielded glycine residues to the pore without significant rotational movement of the associated helices. Together with other hydrophobic amino acids, the phenylalanines act as the channel's gate. Structured M1-M2 turrets, not evident previously, form the channel's extracellular entrance.
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Calcio/metabolismo , Proteínas de Drosophila/metabolismo , Proteína ORAI1/metabolismo , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales , Microscopía por Crioelectrón , Proteínas de Drosophila/genética , Células HEK293 , Humanos , Fragmentos Fab de Inmunoglobulinas , Ratones , Modelos Moleculares , Proteína ORAI1/genética , Conformación Proteica , Saccharomycetales/metabolismoRESUMEN
In order to further understand the influence of high temperature shock on the microbial community structure of activated sludge during the process of nitrite oxidation, the enriched nitrifying activated sludge under different NO2--N concentration was taken as the research object in this study. 16S rRNA high-throughput sequencing technology was used to analyze the changes in the microbial community abundance and structural characteristics of activated sludge by changing the environmental temperature. The results of high-throughput sequencing showed that microorganisms were more likely to grow at 25â, and the diversity of the microbial community in the activated sludge was the most abundant. With increased temperature, the richness, evenness, and diversity of the flora in the system decreased. In addition, it was found that the main nitrifying bacterium in the system was Nitrospira of Nitrospirae, whereby 35â was more suitable for its growth. Meanwhile, a higher temperature also caused differences in the structure of non-nitrifying functional microorganisms (e.g., Bacteroidetes, Chlorofulexi, Halomonas, and Pseudomonas) in the activated sludge. The results of this study provide some theoretical reference for the investigation of the distribution characteristics of microbial flora during the process of nitrite oxidation under high temperature shock, and can also be used as reference for relevant high temperature shock tests.
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Nitritos , Aguas del Alcantarillado , Reactores Biológicos , Oxidación-Reducción , ARN Ribosómico 16S/genética , TemperaturaRESUMEN
HCC is one of the fastest-rising causes of cancer-related death. Novel therapeutic approaches for treatment are warranted. The goal of this study is to find effective components from Chinese herbal medicines, which is an important alternative source of anticancer medicine. To this end, six different herbs were selected from various traditional literatures. Soxhlet extractor was used to distill the strong polar and weak polar components of each herb. The inhibitive effect of each component was determined using liver cancer cells BEL7404. From total of 12 extractions, it was found that the combined crude lysate of Amana edulis from water and ethanol system had the best efficacy. At the concentration of 0.1 mg/mL, this component has the highest inhibition rate up to 70%. To investigate the underlying molecular reasons, we observed that the component can significantly induce the liver cancer cells apoptosis and retard the cell reproduction at G2/M stage. Verification experiments showed that this component also has apparent inhibitive effects on other liver cancer cells, such as HepG2 and Huh7. On the other hand, it has less effectiveness on another cell line HepaRG, which retains many characteristics of primary human hepatocytes. The results suggested that there might be highly efficient antihepatoma ingredient in the water and ethanol extraction of Amana edulis. The pure substances remain to be isolated and further research on their targets is required.
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The store-operated calcium (Ca2+) channel Orai governs Ca2+ influx through the plasma membrane of many non-excitable cells in metazoans. The channel opens in response to the depletion of Ca2+ stored in the endoplasmic reticulum (ER). Loss- and gain-of-function mutants of Orai cause disease. Our previous work revealed the structure of Orai with a closed pore. Here, using a gain-of-function mutation that constitutively activates the channel, we present an X-ray structure of Drosophila melanogaster Orai in an open conformation. Well-defined electron density maps reveal that the pore is dramatically dilated on its cytosolic side in comparison to the slender closed pore. Cations and anions bind in different regions of the open pore, informing mechanisms for ion permeation and Ca2+ selectivity. Opening of the pore requires the release of cytosolic latches. Together with additional X-ray structures of an unlatched-but-closed conformation, we propose a sequence for store-operated activation.
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Canales de Calcio/genética , Calcio/química , Proteínas de Drosophila/química , Proteína ORAI1/química , Conformación Proteica , Animales , Calcio/metabolismo , Canales de Calcio/metabolismo , Membrana Celular/química , Cristalografía por Rayos X , Proteínas de Drosophila/genética , Drosophila melanogaster/química , Drosophila melanogaster/genética , Retículo Endoplásmico/química , Retículo Endoplásmico/genética , Mutación , Proteína ORAI1/genéticaRESUMEN
OBJECTIVES: To investigate the effects of microwave ablation on T-lymphocyte subsets and cytokines in patients with hepatocellular carcinoma. MATERIAL AND METHODS: Peripheral blood was collected from 45 patients with hepatocellular carcinoma treated by microwave ablation before treatment, one week and one month after treatment. T cells (CD3+, CD4+ and CD8+ cells), CD4+ CD25+ Tregs, and CD16+ CD56+ NK cells were analyzed by flow cytometry. Levels of cytokines (IL-2, IFN-γ, TNF-α, IL-12, IL-4, IL-6, IL-8, and IL-10) were determined by a Luminex 200 analyzer. RESULTS: Compared with before treatment, CD3+ cells, CD4+ cells and IL-12 increased significantly at one month after the microwave ablation treatment, while IL-4, IL-10 decreased significantly. CONCLUSIONS: Microwave ablation could relieve the suppression of immune function caused by tumors, promote the deviation of Th2/Th1, and improve immune dysfunction in patients with hepatocellular carcinoma.
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Carcinoma Hepatocelular/cirugía , Ablación por Catéter/métodos , Citocinas/metabolismo , Neoplasias Hepáticas/cirugía , Microondas , Subgrupos de Linfocitos T/metabolismo , Anciano , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/metabolismo , Femenino , Citometría de Flujo , Humanos , Interleucinas/metabolismo , Células Asesinas Naturales/metabolismo , Masculino , Persona de Mediana Edad , Linfocitos T Reguladores/metabolismoRESUMEN
BACKGROUND: Microwave ablation has been extensively used for eliminating pulmonary tumors; however, it is usually associated with severe pain under local anesthesia. Decreasing the power and shortening the ablation time can help to relieve the pain; however, this leads to incomplete ablation and an increasing recurrence rate. This research aims to employ an artificial pneumothorax to increase both the curative effect and pain relief during the ablation procedure. MATERIAL AND METHODS: From July 2013 to January 2015, nine patients presenting with 10 subpleural lung tumors (age: 44-78 years) with a high possibility of severe pain underwent the artificial pneumothorax during microwave ablation. The pain assessment scores and complications induced by the artificial pneumothorax were recorded and analyzed by a CT scan follow-up. RESULTS: The tumors of the nine patients were eliminated successfully using microwave ablation with artificial pneumothorax under local anesthesia. The pain caused by the ablation was relieved to a great extent with an average rate of 94.66% (range: 63.3%-100%) and all tumors were ablated completely. No severe complications occurred after the operation. CONCLUSIONS: The artificial pneumothorax is a reliable therapy to improve the curative effect of microwave ablation under local anesthesia by relieving the pain of the patients.
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Ablación por Catéter/métodos , Neoplasias Pulmonares/cirugía , Microondas , Manejo del Dolor/métodos , Neumotórax Artificial/métodos , Adulto , Anciano , Femenino , Humanos , Neoplasias Pulmonares/patología , Masculino , Persona de Mediana Edad , Neumotórax Artificial/efectos adversosRESUMEN
OBJECTIVE: This study aims to explore the clinical effectiveness of a combination therapy of transarterial chemoembolization (TACE) and percutaneous microwave coagulation therapy (PMCT) in treating hepatocellular carcinoma (HCC) abutting the diaphragm. MATERIAL AND METHODS: Six cases with HCC were treated with TACE followed by PMCT one month later with the aid of artificial pneumothorax. RESULTS: CT/MRI revealed complete necrosis in the tumor lesions and the treated tumor margins (≥ 5 mm). Serum alpha-fetoprotein (AFP) levels markedly declined in patients who originally had higher serum AFP levels. Postoperative complications such as fever, mild hepatic dysfunction and pleural effusion were alleviated within a short period of time. All patients were closely monitored through follow-up; all patients survived, except for one patient who received a liver transplantation. CONCLUSIONS: As lesions are either invisible or poorly visible in sonography, determining an effective treatment for HCC abutting the diaphragm remains a particular challenge. TACE and PMCT combined therapy with the aid of artificial pneumothorax proved to be an available treatment option.
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Carcinoma Hepatocelular/terapia , Quimioembolización Terapéutica/métodos , Neoplasias Hepáticas/terapia , Microondas/uso terapéutico , Carcinoma Hepatocelular/patología , Terapia Combinada , Diafragma , Estudios de Seguimiento , Humanos , Neoplasias Hepáticas/patología , Imagen por Resonancia Magnética/métodos , Masculino , Persona de Mediana Edad , Neumotórax Artificial/métodos , Complicaciones Posoperatorias/epidemiología , Estudios Retrospectivos , Tomografía Computarizada por Rayos X/métodos , Resultado del Tratamiento , alfa-Fetoproteínas/metabolismoRESUMEN
KIT autophosphorylation caused by mutation of KIT is considered to be a critical mechanism for the oncogenesis of gastrointestinal stromal tumors (GISTs). However, little is known regarding whether stem cell factor (SCF), the KIT ligand, is able to induce the proliferation of GIST cells by activating the wild-type KIT receptor in GISTs. Imatinib, a tyrosine kinase inhibitor, has been demonstrated to be effective as treatment for the majority of GISTs. However, primary resistance to imatinib in GISTs with wild-type KIT and acquired resistance in GISTs with mutant KIT are becoming increasingly significant problems. The aims of this study were to detect the expression and function of SCF in 68 GIST samples, and to explore the relationship between SCF activity and imatinib resistance using immunohistochemical staining and western blot analysis. Results showed abundant expression of SCF in GISTs and demonstrated that SCF is capable of enhancing GIST cell proliferation. Similar to its ineffectiveness in wild-type GISTs, imatinib also failed to inhibit SCF-induced KIT activation in GISTs with mutant KIT. We also found increased SCF expression in GIST cells treated with imatinib. Overall, our results indicated that SCF-induced KIT activation is a novel essential pathway for the proliferation of GISTs. Imatinib was not able to inhibit the activity of SCF, while it promoted the expression of SCF, which may have contributed to acquired imatinib resistance.
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The plasma membrane protein Orai forms the pore of the calcium release-activated calcium (CRAC) channel and generates sustained cytosolic calcium signals when triggered by depletion of calcium from the endoplasmic reticulum. The crystal structure of Orai from Drosophila melanogaster, determined at 3.35 angstrom resolution, reveals that the calcium channel is composed of a hexameric assembly of Orai subunits arranged around a central ion pore. The pore traverses the membrane and extends into the cytosol. A ring of glutamate residues on its extracellular side forms the selectivity filter. A basic region near the intracellular side can bind anions that may stabilize the closed state. The architecture of the channel differs markedly from other ion channels and gives insight into the principles of selective calcium permeation and gating.
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Canales de Calcio/química , Calcio/química , Proteínas de Drosophila/química , Proteínas de la Membrana/química , Animales , Sitios de Unión , Cristalografía por Rayos X , Proteínas de Drosophila/agonistas , Ácido Glutámico/química , Proteínas de la Membrana/agonistas , Proteína ORAI1 , Porosidad , Unión Proteica , Estructura Secundaria de Proteína , Estructura Terciaria de ProteínaRESUMEN
AIM: To clarify the biological role of stem cell factor (SCF)-mediated wild-type KIT receptor activation in gastrointestinal stromal tumor (GIST) growth. METHODS: The co-expression of wild-type KIT receptor and SCF was evaluated in 51 GIST samples using mutation analysis and immunohistochemistry, and the results were correlated with clinicopathological parameters, including the mitotic count, proliferative index (Ki-67 immunohistochemical staining), mitotic index (phospho-histone H3 immunohistochemical staining) and apoptotic index (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling). Using primary cultured GIST cells, the effect of SCF-mediated wild-type KIT receptor activation was determined by western blotting, methyl thiazolyl tetrazolium (MTT), and apoptosis assays. RESULTS: We found that wild-type KIT receptor and SCF protein were expressed in 100% and 76.5% of the 51 GIST samples, respectively, and the co-expression of wild-type KIT receptor and SCF was associated with known indicators of poor prognosis, including larger tumor size (P = 0.0118), higher mitotic count (P = 0.0058), higher proliferative index (P = 0.0012), higher mitotic index (P = 0.0282), lower apoptosis index (P = 0.0484), and increased National Institutes of Health risk level (P = 0.0012). We also found that the introduction of exogenous SCF potently increased KIT kinase activity, stimulated cell proliferation (P < 0.01) and inhibited apoptosis (P < 0.01) induced by serum starvation, while a KIT immunoblocking antibody suppressed proliferation (P = 0.01) and promoted apoptosis (P < 0.01) in cultured GIST cells. CONCLUSION: SCF-mediated wild-type KIT receptor activation plays an important role in GIST cell growth. The inhibition of SCF-mediated wild-type KIT receptor activation may prove to be particularly important for GIST therapy.
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Neoplasias Gastrointestinales/metabolismo , Tumores del Estroma Gastrointestinal/metabolismo , Proteínas Proto-Oncogénicas c-kit/metabolismo , Factor de Células Madre/metabolismo , Adulto , Anciano , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Femenino , Neoplasias Gastrointestinales/genética , Neoplasias Gastrointestinales/patología , Tumores del Estroma Gastrointestinal/genética , Tumores del Estroma Gastrointestinal/patología , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Índice Mitótico , Mutación , Proteínas Proto-Oncogénicas c-kit/efectos de los fármacos , Proteínas Proto-Oncogénicas c-kit/genética , Factor de Células Madre/farmacología , Células Tumorales CultivadasRESUMEN
OBJECTIVE: To explore the effect of malignant transformation of the L839P, a new mutation site of the PDGFRA gene, on the pathogenesis of gastrointestinal stromal tumors. METHODS: All recombinant plasmids were stably transfected into CHO cells by liposomes. Western blotting was used to detect the expression of PDGFRA protein. The cell growth curve was plotted by cell counting. Flow cytometry was used to detect the cell cycle and apoptosis of CHO cell, respectively. The stably transformed cells were inoculated subcutaneously into the back of nude mice and the mice were used to observe the tumorigenesis. Transient transfection of the mutant-type plasmids of PDGFRA gene and the wild-type plasmids of kit gene into the CHO cells was performed. Western blot was used to detect the expression of kit protein and its phosphorylated forms. RESULTS: PDGFRA protein expressed in the negative control, experimental group and positive control, except the empty vector. The growth curve showed that it was accelerated in the experimental group and positive control. The ratios of cells in proliferative phase were 28.4% (blank), 24.5% (negative control), 43.8% (experimental group) and 40.9% (positive control). Their apoptotic indexes were 1.8%, 1.9%, 1.5% and 1.6%, respectively. After three weeks, tumors were observed in the nude mice of experimental group and positive control, inoculated with the stably transformed cells. Moreover, the expression of phosphorylated protein of kit was enhanced after cotransfection of the mutant-type plasmids of PDGFRA and the wild-type plasmid of kit. CONCLUSION: The PDGFRA mutant L839P is a gain-of-function mutation and has obviously malignant transforming effect on normal cells, and may activate kit protein accelerating the tumorigenesis. Gastrointestinal stromal tumors;
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Transformación Celular Neoplásica , Tumores del Estroma Gastrointestinal/genética , Mutación , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/genética , Animales , Apoptosis , Células CHO , Ciclo Celular , Proliferación Celular , Cricetinae , Cricetulus , Tumores del Estroma Gastrointestinal/etiología , Tumores del Estroma Gastrointestinal/patología , Ratones , Ratones Desnudos , Plásmidos , Proteínas Proto-Oncogénicas c-kit/metabolismo , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/metabolismo , TransfecciónRESUMEN
PURPOSE: This study presents a new rat oral implant model for assessing histologic changes in the mechanical environment surrounding loaded and unloaded dental implants. MATERIALS AND METHODS: The maxillary left first molar from retired breeder rats was extracted, and the site was allowed to heal for 1 month. A titanium miniscrew implant was then placed into the site and allowed to heal for 21 days. The mandibular left first molars in one group of rats were extracted to create an unloaded condition; in a second group of rats the mandibular left first molars were left in occlusion with the opposing screw head to simulate loading. Radiographs were taken on the day of placement and again at 7 days, 14 days, and 21 days after placement and were used to estimate the bone-implant contact ratio. The rats were sacrificed after 21 days. Peri-implant tissue samples from day 21 were processed for histology and immunohistochemistry with antibodies to osteocalcin and matrix metalloproteinase 13 (MMP-13). Two-dimensional finite element models were created from images of the histologic sections and immunohistochemical samples to observe tissue changes. RESULTS: Areas of high shear stress adjacent to the helical threads of loaded implants were associated with osteocalcin localization and bone formation but only minimal localization of MMP-13. Bone adjacent to unloaded implants showed fibrous tissue and extensive MMP-13 localization surrounding the apical two-thirds of each implant. These results agree with estimated bone-implant contact ratios, which showed a steady decrease in contact ratio for the unloaded implant group but a significantly higher contact ratio in the loaded group between 14 and 21 days. CONCLUSION: The rat oral implant model is useful for studies of the mechanical and physiologic environment affecting osseointegration in loaded and unloaded implants.
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Pilares Dentales , Implantes Dentales , Materiales Dentales , Maxilar/patología , Titanio , Animales , Fenómenos Biomecánicos , Fuerza de la Mordida , Materiales Dentales/química , Femenino , Análisis de Elementos Finitos , Masculino , Metaloproteinasa 13 de la Matriz/análisis , Maxilar/diagnóstico por imagen , Maxilar/cirugía , Modelos Animales , Diente Molar/cirugía , Oseointegración/fisiología , Osteocalcina/análisis , Osteoclastos/patología , Osteogénesis/fisiología , Tejido Periapical/enzimología , Tejido Periapical/patología , Radiografía , Ratas , Ratas Sprague-Dawley , Estrés Mecánico , Factores de Tiempo , Titanio/química , Extracción Dental , Alveolo Dental/diagnóstico por imagen , Alveolo Dental/patología , Alveolo Dental/cirugíaRESUMEN
Bisphosphonates such as alendronate (ALD), although controversial, are worthy of investigation for the enhancement of implant osseointegration in patients with low bone mass who are already taking bisphosphonates for osteoporosis. These patients may receive additional benefits and be acceptable candidates for dental implants without needing to change their medication regimen and possibly as a result of their medication regimen. The purpose of this study was to compare implant osseointegration in maxillary bone of normal rats with a rat model of postmenopausal estrogen deficiency (ovariectomized [OVX]), with and without ALD. An experimental group of 32 rats was divided in 4 groups: ALD-OVX (n=8 OVX with ALD), OVX (n=8 OVX without ALD), ALD (n=8 normal rats with ALD), and control (n=8 normal rats). All rats received one titanium microscrew implant in the left edentulous region of the maxillary arch. The ALD-OVX and ALD groups received subcutaneous injections of ALD 3 times a week. On the fourth week after ALD administration, an implant was placed in all 32 rats. The maxilla of each rat was radiographed 4 times: at 0, 7, 14, and 28 days. On day 28 after implant placement, all rats were killed, and the peri-implant tissue was embedded in plastic or paraffin for histological examination. The X rays were used for a chronologic calculation of the contact ratio between implant and bone surfaces. Radiographic bone density was determined at 3 points: mesial, apical, and distal. The results show that osseointegration of the implants was impaired in the estrogen-deficient OVX rats compared with the ALD-OVX rats. Fifty percent of the implants were lost at 2 weeks in the OVX group. Radiographic evidence suggested that none of the implants in the OVX group osseointegrated. In the histologic examination more bone was observed around implants from the ALD-OVX and ALD groups than around implants from the OVX group. The OVX group presented a dramatic reduction in implant bone contact at 2 weeks and a significant 13% reduction at 4 weeks vs day of implant (P = .006). The ALD-OVX group presented 50% more bone density than the OVX group (P = .0003). Both ALD groups (ALD and ALD-OVX) had significantly higher radiographic bone density than the other groups (P < .01 for each comparison). In conclusion, osseointegration of implants was enhanced by ALD. Radiographic bone density and contact ratio improved with ALD administration. Implant osseointegration was impaired by estrogen deficiency in the OVX group.
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Alendronato/farmacología , Conservadores de la Densidad Ósea/farmacología , Implantes Dentales , Estrógenos/deficiencia , Oseointegración , Alendronato/administración & dosificación , Análisis de Varianza , Animales , Densidad Ósea , Conservadores de la Densidad Ósea/administración & dosificación , Implantación Dental Endoósea , Modelos Animales de Enfermedad , Femenino , Inyecciones Subcutáneas , Maxilar/cirugía , Oseointegración/efectos de los fármacos , Ovariectomía , Ovario/fisiología , Ratas , Ratas Sprague-DawleyRESUMEN
Snake secreted phospholipasesA2 (sPLA2s) are widely used as pharmacological tools to investigate their role in diverse pathophysiological processes. Some members of snake venom sPLA2s have been found to block voltage-activated K(+) channels (K(v) channels). However, most studies involved in their effects on ion channels were indirectly performed on motor nerve terminals while few studies were directly done on native neurons. Here, a novel snake sPLA2 peptide neurotoxin, Natratoxin, composed of 119 amino acid residues and purified from Naja atra venom was reported. It was characterized using whole-cell patch-clamp in acutely dissociated rat dorsal root ganglion (DRG) neurons. It was found to effectively inhibit A-type K(+) currents and cause alterations of channel gating characters, such as the shifts of steady-state activation and inactivation curves to hyperpolarization direction and changes of V(1/2) and slope factor. Therefore, Natratoxin was suggested to be a gating modifier of K(v) channel. In addition, this inhibitory effect was found to be independent of its enzymatic activity. These results suggested that the toxin enacted its inhibitory effect by binding to K(v) channel. To further elucidate the structural basis for this electrophysiological phenomenon, we determined the crystal structure of Natratoxin at 2.2 A resolution by molecular replacement method and refined to an R-factor of 0.190. The observed overall fold has a different structural organization from other K(+) channel inhibitors in animal toxins. Compared with other K(v) channel inhibitors, a similar putative functional surface in its C-terminal was revealed to contribute to protein-protein interaction in such a blocking effect. Our results demonstrated that the spatial distribution of key amino acid residues matters most in the recognition of this toxin towards its channel target rather than its type of fold.
Asunto(s)
Venenos Elapídicos/química , Fosfolipasas A2/farmacología , Bloqueadores de los Canales de Potasio/farmacología , Canales de Potasio/efectos de los fármacos , Animales , Cristalografía por Rayos X , Venenos Elapídicos/farmacología , Elapidae , Cobayas , Técnicas de Placa-Clamp , Fosfolipasas A2/química , Conformación Proteica , Ratas , Ratas WistarRESUMEN
MitoNEET was identified as an outer mitochondrial membrane protein that can potentially bind the anti-diabetes drug pioglitazone. The crystal structure of the cytoplasmic mitoNEET (residues 33-108) is determined in this study. The structure presents a novel protein fold and contains a [2Fe-2S] cluster-binding domain. The [2Fe-2S] cluster is coordinated to the protein by Cys-72, Cys-74, Cys-83, and His-87 residues. This coordination is also novel compared with the traditional [2Fe-2S] cluster coordinated by four cysteines or two cysteines and two histidines. The cytoplasmic mitoNEET forms homodimers in solution and in crystal. The dimerization is mainly mediated by hydrophobic interactions as well as hydrogen bonds coordinated by two water molecules binding at the interface. His-87 residue, which plays an important role in the coordination of the [2Fe-2S] cluster, is exposed to the solvent on the dimer surface. It is proposed that mitoNEET dimer may interact with other proteins via the surface residues in close proximity to the [2Fe-2S] cluster.