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A self-sufficient cytochrome P450 monooxygenase from Deinococcus apachensis (P450DA) was identified and successfully overexpressed in Escherichia coli BL21(DE3). P450DA would be a member of the CYP102D subfamily and assigned as CYP102D2 according to the phylogenetic tree and sequence alignment. Purification and characterization of the recombinant P450DA indicated both NADH and NADPH could be used by P450DA as a reducing cofactor. The recombinant E.â coli (P450DA) strain was functionally active, showing excellent enantioselectivity for benzylic hydroxylation of methyl 2-phenylacetate. Further substrate scope studies revealed that P450DA is able to catalyze benzylic hydroxylation of a variety of compounds, affording the corresponding chiral benzylic alcohols in 86-99 %â ee and 130-1020 total turnover numbers.
Assuntos
Proteínas de Bactérias/metabolismo , Álcool Benzílico/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Deinococcus/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/classificação , Proteínas de Bactérias/genética , Álcool Benzílico/química , Biocatálise , Sistema Enzimático do Citocromo P-450/classificação , Sistema Enzimático do Citocromo P-450/genética , Escherichia coli/metabolismo , Hidroxilação , NADP/química , NADP/metabolismo , Filogenia , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Estereoisomerismo , Especificidade por SubstratoRESUMO
Lung cancer is the most common cancer and leading cause of cancer-related deaths worldwide. The first-generation reversible, ATP-competitive inhibitors gefetinib and elotinib showed good clinical responses in lung adenocarcinoma tumors (NSCLC). But almost all patients developed resistance to these inhibitors over time. Such resistance of EGFR inhibitors was frequently linked to the acquired L858R and T790M point mutations in the kinase domain of EGFR. To overcome these resistance problems, the second and the third generation inhibitors have been discovered. FDA approved afatinib, the second generation irreversible inhibitor and osimitinib, the third generation irreversible EGFR inhibitors for the treatments of NSCLC. We identified new covalent quinazoline inhibitors (E)-N-(4-(3-chloro-4-fluorophenylamino)-7-(2-ethoxyethoxy)quinazolin-6-yl)-4-(dimethylamino)but-2-enamide (6d) and (E)-N-(4-(3-chloro-4-(pyridin-2-ylmethoxy)phenylamino)-7-(2-ethoxyethoxy)quinazolin-6-yl)-4-(dimethyl-amino)but-2-enamide (6h) that exhibited potent EGFR kinase inhibitory activities on L858R and T790M mutations. The compound 6 h showed selectivity similar to AZD9291 (osimertinib) in mutated and wild type tumor cell lines. In vitro cell assay 6d and 6h were better than afatinib and osimertinib. In vivo antitumor efficacy studies of these compounds were done in NCI-H1975 mice xenografts.
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Antineoplásicos/farmacologia , Modelos Animais de Doenças , Inibidores de Proteínas Quinases/farmacologia , Quinazolinas/farmacologia , Receptor ErbB-2/antagonistas & inibidores , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Receptores ErbB/antagonistas & inibidores , Receptores ErbB/metabolismo , Feminino , Humanos , Neoplasias Hepáticas Experimentais/tratamento farmacológico , Neoplasias Hepáticas Experimentais/metabolismo , Neoplasias Hepáticas Experimentais/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Estrutura Molecular , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/química , Quinazolinas/síntese química , Quinazolinas/química , Receptor ErbB-2/metabolismo , Relação Estrutura-AtividadeRESUMO
Overexpression of EGFR and HER2 are observed in many breast, ovarian, colon and prostate cancers. The second and third generation irreversible EGFR/HER2 dual kinase inhibitors became popular after the approval of Afatinib by FDA to overcome the mutation related problem. To find efficacious drug candidates, a series of novel quinazoline derivatives were designed, synthesized and evaluated as dual EGFR/HER2 tyrosine kinase (TK) inhibitors. Selected twenty four compounds were reported here with significant inhibitory activities against EGFR/HER2 tyrosine kinases. Several compounds showed nanomolar IC50 values. In vitro studies of quinazoline derivatives were done on NCI-H1975, HCC827, A431, MDA MB-453 cell lines. The compounds 1a, 1d and 1v were found more potent compared to standard drug afatinib. In vivo efficacy study of 1d on nude mice NCI-H1975 tumour xenograft model was discussed.
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Antineoplásicos/farmacologia , Descoberta de Drogas , Receptores ErbB/antagonistas & inibidores , Quinazolinas/farmacologia , Receptor ErbB-2/antagonistas & inibidores , Animais , Linhagem Celular Tumoral , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
CO2/CH4 interaction determines the prospects for complementary enhanced gas recovery (EGR) associated with CO2 sequestration in shale. We characterize the competitive adsorption of CO2 and CH4 in shale using low-field NMR. Competitive sorption of CO2 relative to CH4 is defined as the CO2/CH4 competitive adsorption ratio (CO2/CH4 CAR for short) when CO2 and CH4 have the same original partial pressure in shale. Results indicate the CO2/CH4 CAR decreases with the logarithm of increasing pressure. Observed CO2/CH4 CARs are on the order of 4.28-5.81 (YDN-1) to 3.43-5.57 (YDN-2), describing the remarkable competitive advantage of CO2 sorption relative to CH4 for shale. Results also indicate that increasing the CO2/CH4 pressure ratio (1) increases the adsorption capacity of shales to CO2 and decreases that to CH4 logarithmically with pressure, and (2) boosts CO2-CH4 displacement and generates greater EGR efficiency in shale, where the EGR efficiency can be inferred by the CO2/CH4 pressure ratio using a Langmuir-like function. Furthermore, the maximum sequestration capacity of adsorbed CO2 during CO2-CH4 competition is on the order of â¼3.87 cm3/g (YDN-1) to â¼5.13 cm3/g (YDN-2). These promising results for EGR and CO2 storage reveal the considerable potential for carbon capture and geological sequestration in shale.
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Dióxido de Carbono , Carbono , Adsorção , Geologia , Metano , MineraisRESUMO
Gel plugging agents have become one of the preferred methods for plugging in complex and severe loss conditions during drilling due to their good adaptability to loss channels. To address the common issue of poor temperature resistance in gel-based plugging agents, high-temperature-resistant gel plugging materials were synthesized through the molecular design of polymers, modifying existing agents. Based on the temperature and salt resistance of the aqueous solution of an acrylamide (AM)/N-vinylpyrrolidone (NVP) binary copolymer, temperature-resistant monomer sodium styrene sulfonate (SSS) was introduced and reacted in a polyvinyl alcohol (PVA) aqueous solution. Using ammonium persulfate (APS) as an initiator and crosslinking with N,N-methylenebisacrylamide (MBA), a gel plugging material resistant to 140 °C was synthesized. The structure, thermal stability, water absorption and expansion, and plugging performance of the gel were studied through hot rolling aging, thermogravimetric analysis, infrared spectroscopy, electron microscopy scanning, sand bed experiments, and drag reduction experiments. The results show that the gel material has good thermal stability and water absorption and expansion at 140 °C, and its temperature-resistant plugging performance is excellent, significantly slowing down the loss rate of drilling fluid. This provides a basis for the further development of gel materials.
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Neonicotinoids are ubiquitous in global surface waters and pose a significant risk to aquatic organisms. However, information is lacking on the variations in sensitivity of organisms at different developmental stages to the neurotoxic neonicotinoids. We established a spectrum of toxicity to zebrafish embryos at four neurodevelopmental stages (1, 3, 6, and 8 h post fertilization [hpf]) and dechorionated embryos at 6 hpf based on external and internal exposure to imidacloprid as a representative neonicotinoid. Embryos at the gastrula stage (6 and 8 hpf) were more sensitive to imidacloprid than embryos at earlier developmental stages. Dechorionated embryos were more sensitive to imidacloprid than embryos with a chorion, suggesting that the chorion offers protection against pollutants. Nine sublethal effects were induced by imidacloprid exposure, among which uninflated swim bladder (USB) was the most sensitive. Water depth and air availability in the exposure chambers were critical factors influencing the occurrence of USB in zebrafish larvae. Internal residues of metabolites accounted for <10% of imidacloprid, indicating that imidacloprid was metabolized in a limited fashion in the embryos. In addition, acute toxicity of the main metabolite 5-hydroxy-imidacloprid was significantly lower than that of imidacloprid, indicating that the observed toxicity in embryos exposed to imidacloprid was mainly induced by the parent compound. Our research offers a fresh perspective on choosing the initial exposure time in zebrafish embryo toxicity tests, particularly for neurotoxicants. Environ Toxicol Chem 2024;00:1-11. © 2024 SETAC.
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The requirement for cryogenic supramolecular self-assembly of amphiphiles in subzero environments is a challenging topic. Here, the self-assembly of lamellar lyotropic liquid crystals (LLCs) are presented to a subzero temperature of -70 °C. These lamellar nanostructures are assembled from specifically tailored ultra-long-chain surfactant stearyl diethanolamine (SDA) in water/glycerol binary solvent. As the temperature falls below zero, LLCs with a liquid-crystalline Lα phase, a tilted Lß phase, and a new folded configuration are obtained consecutively. A comprehensive experimental and computational study is performed to uncover the precise microstructure and formation mechanism. Both the ultra-long alkyl chain and head group of SDA play a crucial role in the formation of lamellar nanostructures. SDA head group is prone to forming hydrogen bonds with water, rather than glycerol. Glycerol cannot penetrate the lipid layer, which mixes with water arranging outside of the lipid bilayer, providing an ideal anti-freezing environment for SDA self-assembly. Based on these nanostructures and the ultra-low freezing point of the system, a series of novel cryogenic materials are created with potential applications in extremely cold environments. These findings would contribute to enriching the theory and research methodology of supramolecular self-assembly in extreme conditions and to developing novel anti-freezing materials.
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Damaged adobe masonry is essential to retrofit not only for continued use by less developed populations but also for historical preservation and vernacular landmark maintenance. Coating mortar on adobe can greatly improve the static load-carrying capacity of adobe masonry Wallette. Four retrofitting methods are carried out to enhance the cooperation of mortar and adobe; these four methods focus on the factors of roughness, shear dowel, and mesh size and a novel biological and traditional Chinese binder, sticky rice pulp. The mortar coating with the four bonding enhancement methods can greatly increase both the compressive and lateral load-carrying capacities, with a maximum improvement of 177 and 743%, respectively. The bonding strength has a negative effect on the compressive load-carrying capacity; on the other hand, it has a positive effect on the lateral load-carrying capacity. A range analysis is also carried out, which shows that the shear dowel depth has the greatest effect on both load situations, followed by the wire mesh size.
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The pyrolysis process of source rock, especially organic-rich immature shale, is required for oil and gas extraction, during which the evolution of the pore structure system in the immature shale determines the heat conduction and fluid flow under the heating treatment. Although some sound achievements have been made regarding the pyrolysis of immature shale, the effect of the total organic carbon (TOC) content on the pore structure evolution of immature shale remains unclear. With respect to this issue, in this work, a series of N2 adsorption/desorption and nuclear magnetic resonance (NMR) experiments were conducted, and fractal dimension theory was also introduced to analyze the pore structure evolution of immature shale subjected to heating treatment in a quantitative manner. The results indicate that the adsorption branch of the nitrogen adsorption-desorption isotherm can be divided into three stages. The pore structure of different TOC immature shales does not change significantly, and they are all slit-shaped. In addition, immature shale with a higher organic content has a higher hydrocarbon expulsion strength and a higher pore volume growth rate, which indicate that the pyrolysis of organic matter greatly affects the pore structure of immature shale during heating. This phenomenon shows that the pyrolysis of organic matter greatly influences the pore structure of immature shale during the heating process. The pores of immature shale in the study area have significant fractal characteristics, the fractal dimension is between 2.397 and 2.636, the pore space of the sample is extremely small, the pore structure is extremely complex, and the heterogeneity is strong.
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Prostaglandin E2 (PGE2) receptor 4 (EP4) is one of four EP receptors commonly upregulated in the tumor microenvironment and plays vital roles in stimulating cell proliferation, invasion, and metastasis. Biochemical blockade of the PGE2-EP4 signaling pathway is a promising strategy for controlling inflammatory and immune related disorders. Recently combination therapies of EP4 antagonists with anti-PD-1 or chemotherapy agents have emerged in clinical studies for lung, breast, colon, and pancreatic cancers. Herein, a novel series of indole-2-carboxamide derivatives were identified as selective EP4 antagonists, and SAR studies led to the discovery of the potent compound 36. Due to favorable pharmacokinetics properties and good oral bioavailability (F = 76%), compound 36 was chosen for in vivo efficacy studies. Compound 36 inhibited tumor growth in a CT-26 colon cancer xenograft better than E7046 and a combination of 36 with capecitabine significantly suppressed tumor growth (TGI up to 94.26%) in mouse models.
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The main advantage of having livestock, for example, the laying hens, in a controlled environment is that the optimum growth conditions can be achieved with accuracy. The indoor air temperature, humidity, gases concentration, etc., would significantly affect the animal performance, thus should be maintained within an acceptable range. In order to achieve the goals of precision poultry farming, various models have been developed by researchers all over the world to estimate the hourly indoor environmental parameters so as to provide decision suggestions. However, a key parameter of hourly manure area in the poultry house was missing in the literature to predict the ammonia emission using the recently developed mechanistic model. Therefore, in order to fill the gap of the understanding of hourly manure coverage proportion and area on the manure belt, experimental measurements were performed in the present study using laying hens from 10 weeks age to 30 weeks age. For each test, six polypropylene (pp) plates were applied to collect the manure dropped by the birds every hour, and photographs of the plates were taken at the same time using a pre-fixed camera. Binary images were then produced based on the color pictures to determine the object coverage proportion. It was demonstrated that for laying hens of stocking density around 14 birds/m2, the manure coverage proportion at the 24th hour after the most recent manure removal was about 60%, while the value was approximately 82% at the 48th hour. Meanwhile, for laying hens at different ages, the hourly increment of manure coverage proportion showed a similar pattern with four distinct stages within 48 h. The statistical analyses demonstrated no significant correlation between the hourly increment of manure weight and the hourly increment of manure coverage proportion. Finally, prediction models for estimating the hourly manure coverage proportion on the manure belt in typical laying hen houses were provided.
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Despite recent successes in the control of dental caries, the mechanism of caries development remains unclear. To investigate the causes of dental decay, especially in early childhood caries, the supragingival microflora composition of 20 twins with discordant caries phenotypes were analyzed using high-throughput pyrosequencing. In addition, the parents completed a lifestyle questionnaire. A total of 228,789 sequencing reads revealed 10 phyla, 84 genera, and 155 species of microflora, the relative abundances of these strains varied dramatically among the children, Comparative analysis between groups revealed that Veillonella, Corynebacterium and Actinomyces were presumed to be caries-related genera, Fusobacterium, Kingella and Leptotrichia were presumed to be healthy-related genus, yet this six genera were not statistically significant (P>0.05). Moreover, a cluster analysis revealed that the microbial composition of samples in the same group was often dissimilar but that the microbial composition observed in twins was usually similar. Although the genetic and environmental factors that strongly influence the microbial composition of dental caries remains unknown, we speculate that genetic factors primarily influence the individual's susceptibility to dental caries and that environmental factors primarily regulate the microbial composition of the dental plaque and the progression to caries. By using improved twins models and increased sample sizes, our study can be extended to analyze the specific genetic and environmental factors that affect the development of caries.
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Bactérias/classificação , Bactérias/isolamento & purificação , Cárie Dentária/microbiologia , Placa Dentária/microbiologia , Doenças em Gêmeos/microbiologia , Análise de Sequência de DNA/métodos , Actinomyces/isolamento & purificação , Criança , Pré-Escolar , Corynebacterium/isolamento & purificação , DNA Ribossômico/genética , Cárie Dentária/patologia , Feminino , Fusobacterium/isolamento & purificação , Humanos , Masculino , RNA Ribossômico 16S/genética , Fatores de Risco , Veillonella/isolamento & purificaçãoRESUMO
BACKGROUND: Randomized controlled trials (RCTs) provide the highest level of evidence and are likely to influence clinical decision-making. This study evaluated the reporting quality of RCT abstracts on drug therapy of periodontal disease and assessed the associated factors. MATERIAL AND METHODS: The Pubmed database was searched for periodontal RCTs published in Science Citation Indexed (SCI) dental journals from 2010/01/01 to 2019/07/17. Information was extracted from the abstracts according to a modified Consolidated Standards of Reporting Trials (CONSORT) guideline checklist. The data was analyzed using descriptive statistical analysis and the statistical associations were examined using the linear regression analysis (P < 0.05). RESULTS: This study retrieved 1715 articles and 249 of them were finally included. The average overall CONSORT score was 15.6 ± 3.4, which represented 40.9% (±0.6) of CONSORT criteria filling. The reporting rate of some items (trial design, numbers analyzed, confidence intervals, intention-to-treat analysis or per-protocol analysis, harms, registration) was less than 30%. The adequate reporting rate of some items (participants, randomization, numbers analyzed, confidence intervals, intention-to-treat analysis or per protocol analysis) was no more than 4%. None of the abstracts reported funding. According to the multivariable linear regression results, number of authors (P = 0.030), word count (P < 0.001), continent (P = 0.003), structured format (P < 0.001), type of periodontal disease (P < 0.001) and international collaboration (P = 0.023) have a significant association with reporting quality. CONCLUSIONS: The quality of RCT abstracts on drug therapy of periodontal disease in SCI dental journals remained suboptimal. More efforts should be made to improve RCT abstracts reporting quality
No disponible
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Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto/normas , Indexação e Redação de Resumos/normas , Doenças Periodontais/tratamento farmacológico , Modelos Lineares , Análise Multivariada , Intervalos de Confiança , Bases de Dados Bibliográficas/normasRESUMO
Ni(II) dihalides bearing two different or identical NHC ligands have been prepared via a controlled indene elimination synthesis, and the former product provides a new route for the design of biscarbene Ni(II)-based catalysts. The indene elimination reaction of the indenynickel(II) complex (1-H-Ind)Ni(NHC)X (Ind = indenyl) with one equiv. of a distinct imidazolium salt at 100 °C afforded the first example of Ni(II) dihalides bearing two different NHC ligands, i.e., Ni(iPr)(IPr)X(2) [iPr = 1,3-diisopropylimidazol-2-ylidene, IPr = 1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene), X = Cl, 1; X = Br, 2] and Ni(iPr)(IMes)Br(2) [IMes = 1,3-bis(mesityl)imidazol-2-ylidene, 3]. Alternatively, complexes 1-3 can be synthesized using a bis-indenyl Ni(II) complex (1-H-Ind)(2)Ni as starting materials via a step-by-step indene elimination at different reaction temperatures. The direct reaction of (1-R-Ind)(2)Ni (R = H or Me) with two equiv. of imidazolium salts at 100 °C afforded Ni(II) dihalides bearing two identical NHC ligands, i.e., Ni(iPr)X(2) (X = Cl, 4; X = Br, 5) and Ni(IPr)Cl(2) (6). All of these complexes were characterized by elemental analysis, NMR spectroscopy and X-ray crystallography for complexes 1-5. The two identical or different NHC ligands in complexes 1-6 changed the coordination sphere of the nickel center from a typical square-planar geometry to a slightly tetrahedral array. A preliminary catalytic study on the cross-coupling reactions of aryl Grignard reagents with aryl halides revealed that complexes 1 and 2 possess the highest activity. In comparison, complexes 3 and 6 exhibited moderate activity and the least active complexes were 4 and 5.