RESUMO
ABSTRACT: Clinical practice shows that a critical unmet need in the field of thrombosis prevention is the availability of anticoagulant therapy without bleeding risk. Inhibitors against FXIa or FXIIa have been extensively studied because of their low bleeding risk. However, whether these compounds produce synergistic effects has not yet been explored. In this study, analyses of activated partial thromboplastin time in combination with the FXIa inhibitor PN2KPI and the FXIIa inhibitor Infestin4 at different proportions were performed using the SynergyFinder tool identifying synergistic anticoagulation effects. Both an FeCl 3 -induced carotid artery thrombosis mouse model and a transient occlusion of the middle cerebral artery mouse model showed that the combination of PN2KPI and Infestin4, which are 28.57% and 6.25% of the effective dose, respectively, significantly prevents coagulation, and furthermore, dual inhibition does not cause bleeding risk.
Assuntos
Anticoagulantes , Coagulação Sanguínea , Modelos Animais de Doenças , Sinergismo Farmacológico , Fator XIIa , Fator XIa , Animais , Fator XIa/antagonistas & inibidores , Fator XIa/metabolismo , Anticoagulantes/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Masculino , Fator XIIa/antagonistas & inibidores , Fator XIIa/metabolismo , Trombose das Artérias Carótidas/prevenção & controle , Trombose das Artérias Carótidas/induzido quimicamente , Trombose das Artérias Carótidas/tratamento farmacológico , Camundongos , Infarto da Artéria Cerebral Média/tratamento farmacológico , Hemorragia/induzido quimicamente , Camundongos Endogâmicos C57BL , Tempo de Tromboplastina ParcialRESUMO
Distance has been one of the basic factors in manufacturing and control fields, and ultrasonic distance sensors have been widely used as a low-cost measuring tool. However, the propagation of ultrasonic waves is greatly affected by environmental factors such as temperature, humidity and atmospheric pressure. In order to solve the problem of inaccurate measurement, which is significant within industry, this paper presents a novel ultrasonic distance sensor model using networked error correction (NEC) trained on experimental data. This is more accurate than other existing approaches because it uses information from indirect association with neighboring sensors, which has not been considered before. The NEC technique, focusing on optimization of the relationship of the topological structure of sensor arrays, is implemented for the compensation of erroneous measurements caused by the environment. We apply the maximum likelihood method to determine the optimal fusion data set and use a neighbor discovery algorithm to identify neighbor nodes at the top speed. Furthermore, we adopt the NEC optimization algorithm, which takes full advantage of the correlation coefficients for neighbor sensors. The experimental results demonstrate that the ranging errors of the NEC system are within 2.20%; furthermore, the mean absolute percentage error is reduced to 0.01% after three iterations of this method, which means that the proposed method performs extremely well. The optimized method of distance measurement we propose, with the capability of NEC, would bring a significant advantage for intelligent industrial automation.
Assuntos
Algoritmos , Artefatos , Redes de Comunicação de Computadores/instrumentação , Transdutores , Ultrassonografia/instrumentação , Ultrassonografia/métodos , Calibragem , Redes de Comunicação de Computadores/normas , Desenho de Equipamento , Análise de Falha de Equipamento , Ultrassonografia/normasRESUMO
A method of direct contact membrane distillation (DCMD) with a self-made hollow polyvinylidene fluoride membrane was applied to prepare high concentration polyaluminum chloride (PACl) with high A1b content based on chemical synthesis. The permeate flux and A1 species distribution were investigated. The experimental results showed that the permeate flux decreased from 14 to 6 kg/(m2 x hr) at the end of the DCMD process, which can be mainly attributed to the formation of NaCl deposits on the membrane surface. The Alb content decreased slightly, only from 86.3% to 84.4%, when the DCMD experiment finished, correspondingly the A1c content increased slightly from 7.2% to 8.5%, and the A1a content remained at 7% during the whole DCMD process. A PACl with A1b content of 84% at total aluminum concentration 2.2 mol/L was successfully prepared by the chemical synthesis-DCMD method.
Assuntos
Hidróxido de Alumínio/síntese química , Destilação/métodos , Membranas Artificiais , Alumínio/química , Hidróxido de Alumínio/química , Condutividade Elétrica , Microscopia Eletrônica de Varredura , Permeabilidade , Polivinil/química , Soluções , Espectrometria por Raios X , Temperatura , Água/químicaRESUMO
AIMS: Anticoagulation in disease treatment has been wildly studied in recent years. The intrinsic coagulation pathway is attracting attention of research community due to its low bleeding risk, and inhibitors against intrinsic coagulation factor XIIa (FXIIa) or XIa (FXIa) have been extensively studied. However, studies to develop anticoagulant inhibitors simultaneous targeting FXIIa and FXIa have not been reported. Our study aimed to evaluate the anticoagulation effect of the dual targeting of FXIIa and FXIa. MAIN METHODS: A fusion protein Infestin-PN2KPI (IP) was designed by linking FXIIa inhibitor Infesin4 and FXIa inhibitor PN2KPI through a rigid linker, and was cloned, expressed and characterized. The binding of IP to FXIIa and FXIa was verified by SPR, and inhibitory ability of IP against FXIIa and FXIa was verified by chromogenic substrate method. And then, the anticoagulation and antithrombotic functions of IP were extensively evaluated by aPTT assay, FeCl3-induced carotid artery thrombosis model and transient occlusion of the middle cerebral artery model. KEY FINDINGS: IP significantly prolonged aPTT, inhibited thrombosis and prevented stroke at a dose of at least 1/2 lower than the effective dose of its component Infestin4 or PN2KPI, and did not cause bleed risk. SIGNIFICANCE: The bifunctional fusion protein IP showed good anticoagulation effects, and simultaneous targeting FXIIa and FXIa is a promising strategy for anticoagulation drug development.
Assuntos
Fator XIIa , Fator XIa , Humanos , Fator XIa/química , Fator XIa/metabolismo , Fibrinolíticos/farmacologia , Compostos Cromogênicos , Anticoagulantes/farmacologia , HemorragiaRESUMO
FXIa inhibition has been a promising strategy for treating thrombotic diseases. Up to date, many small-molecule FXIa inhibitors have been identified; however, most of them exhibit undesirable selectivity over the homologous plasma kallikrein (PKal). By employing structure-based drug design strategies, we identified many novel selective FXIa inhibitors that have extra interactions with the S2 subsite of FXIa. Among them, compound 35 displayed good inhibitory activity against FXIa and high selectivity over PKal and even several other serine proteases. Additionally, 35 showed significant anticoagulant activity toward the intrinsic pathway without affecting the extrinsic pathway. In vivo, 35 exhibited significant antithrombotic activity without increasing the bleeding risk and obvious toxicity in mice, demonstrating that it could be a promising candidate for further research. This study first demonstrates the importance of the S2 subsite of FXIa, paving the way to design highly selective FXIa inhibitors for clinical uses.
Assuntos
Fator XIa , Trombose , Animais , Anticoagulantes/farmacologia , Desenho de Fármacos , Camundongos , Trombose/tratamento farmacológicoRESUMO
Scaffold-based peptides (SBPs) are fragments of large proteins that are characterized by potent bioactivity, high thermostability, and low immunogenicity. Some SBPs have been approved by the FDA for human use. In the present study, we developed SBPs from the venom gland of Deinagkistrodon acutus (D. acutus) by combining transcriptome sequencing and Pfam annotation. To that end, 10 Kunitz peptides were discovered from the venom gland of D. acutus, and most of which peptides exhibited Factor XIa (FXIa) inhibitory activity. One of those, DAKS1, exhibiting strongest inhibitory activity against FXIa, was further evaluated for its anticoagulant and antithrombotic activity. DAKS1 prolonged twofold APTT at a concentration of 15 µM in vitro. DAKS1 potently inhibited thrombosis in a ferric chloride-induced carotid-artery injury model in mice at a dose of 1.3 mg/kg. Furthermore, DAKS1 prevented stroke in a transient middle cerebral-artery occlusion (tMCAO) model in mice at a dose of 2.6 mg/kg. Additionally, DAKS1 did not show significant bleeding risk at a dose of 6.5 mg/kg. Together, our results indicated that DAKS1 is a promising candidate for drug development for the treatment of thrombosis and stroke disorders.
RESUMO
Kunitz-type proteins or peptides have been found in many blood-sucking animals, but the identity of them in leeches remained elusive. In the present study, five Kunitz-type peptides named WPK1-WPK5 were identified from the leech Whitmania pigra. Recombinant WPK1-WPK5 were expressed in Pichia pastoris GS115, and their inhibitory activity against Factor XIa (FXIa) was tested. WPK5 showed inhibitory activity against FXIa with an IC50 value of 978.20 nM. To improve its potency, the loop replacement strategy was used. The loop 1 (TGPCRSNLER) and loop 2 (QYGGC) in WPK5 were replaced by loop 1 (TGPCRAMISR) and loop 2 (FYGGC) in PN2KPI, respectively, and the resulting peptide named WPK5-Mut showed an IC50 value of 8.34 nM to FXIa, which is about 100-fold the potency of FXIa compared to that of WPK5. WPK5-Mut was further evaluated for its extensive bioactivity in vitro and in vivo. It dose-dependently prolonged APTT on both murine plasma and human plasma, and potently inhibited FeCl3-induced carotid artery thrombosis in mice at a dose of 1.5 mg/kg. Additionally, WPK5-Mut did not show significant bleeding risk at a dose of 6 mg/kg. Together, these results showed that WPK5-Mut is a promising candidate for the development of an antithrombotic drug.
RESUMO
The hydrolysis/precipitation behaviors of Al(3+), Al(13) and Al(30) under conditions typical for flocculation in water treatment were investigated by studying the particulates' size development, charge characteristics, chemical species and speciation transformation of coagulant hydrolysis precipitates. The optimal pH conditions for hydrolysis precipitates formation for AlCl(3), PAC(Al13) and PAC(Al30) were 6.5-7.5, 8.5-9.5, and 7.5-9.5, respectively. The precipitates' formation rate increased with the increase in dosage, and the relative rates were AlCl(3)>>PAC(Al30)>PAC(Al13). The precipitates' size increased when the dosage increased from 50 microM to 200 microM, but it decreased when the dosage increased to 800 microM. The Zeta potential of coagulant hydrolysis precipitates decreased with the increase in pH for the three coagulants. The iso-electric points of the freshly formed precipitates for AlCl(3), PAC(Al13) and PAC(Al30) were 7.3, 9.6 and 9.2, respectively. The Zeta potentials of AlCl(3) hydrolysis precipitates were lower than those of PAC(Al13) and PAC(Al30) when pH>5.0. The Zeta potential of PAC(Al30) hydrolysis precipitates was higher than that of PAC(Al13) at the acidic side, but lower at the alkaline side. The dosage had no obvious effect on the Zeta potential of hydrolysis precipitates under fixed pH conditions. The increase in Zeta potential with the increase in dosage under uncontrolled pH conditions was due to the pH depression caused by coagulant addition. Al-Ferron research indicated that the hydrolysis precipitates of AlCl(3) were composed of amorphous Al(OH)(3) precipitates, but those of PAC(Al13) and PAC(Al30) were composed of aggregates of Al(13) and Al(30), respectively. Al(3+) was the most un-stable species in coagulants, and its hydrolysis was remarkably influenced by solution pH. Al(13) and Al(30) species were very stable, and solution pH and aging had little effect on the chemical species of their hydrolysis products. The research method involving coagulant hydrolysis precipitates based on Al-Ferron reaction kinetics was studied in detail. The Al species classification based on complex reaction kinetic of hydrolysis precipitates and Ferron reagent was different from that measured in a conventional coagulant assay using the Al-Ferron method. The chemical composition of Al(a), Al(b) and Al(c) depended on coagulant and solution pH. The Al(b) measured in the current case was different from Keggin Al(13), and the high Al(b) content in the AlCl(3) hydrolysis precipitates could not used as testimony that most of the Al(3+) was converted to highly charged Al(13) species during AlCl(3) coagulation.
Assuntos
Compostos de Alumínio/química , Polímeros/química , Precipitação Química , HidróliseRESUMO
Red mud is the waste of alumina industry and has high TiO2 and Fe2O3 content which are active components for the adsorption of anion pollutants. In this study, the uptake of phosphate by red mud activated by heat treatment and acid-heat treatment was investigated. The factors influencing the adsorption were also investigated. The result showed that the red mud sample treated using acid-heat method at 80 degrees C with 0.25 mol/L HCl for 2 h achieved the highest phosphate removal. For the heat-activated red mud, the sample heated at 700 degrees C for 2 h preformed better than the other heat treatment. Phosphate removal by the activated red mud was significantly pH dependent, and pH 7 was the optimal pH for phosphate removal. The adsorption fits Langmuir isotherm model well and the maximum adsorption capacities of the acid-heat activated red mud and the heat activated samples were 202.9 mgP/g and 155.2 mgP/g, respectively.
Assuntos
Fosfatos/química , Poluentes Químicos da Água/química , Purificação da Água/métodos , Adsorção , Óxido de Alumínio/química , Temperatura Alta , Concentração de Íons de Hidrogênio , Óxido de Magnésio/química , Óxidos/química , Fosfatos/isolamento & purificação , Compostos de Potássio/química , Reprodutibilidade dos Testes , Dióxido de Silício/química , Compostos de Sódio/química , Fatores de Tempo , Titânio/química , Poluentes Químicos da Água/isolamento & purificaçãoRESUMO
The effect of acidification and heat treatment of raw red mud (RM) and fly ash (FA) on the sorption of phosphate was studied in parallel experiments. The result shows that a higher efficiency of phosphate removal was acquired by the activated samples than by the raw ones. The sample prepared by using the RM stirred with 0.25 M HCl for 2h (RM0.25), as well as another sample prepared by heating the RM at 700 degrees C for 2h (RM700), registered the maximum removal of phosphate (99% removal of phosphate). This occurred when they were used in the phosphate sorption studies conducted at pH 7.0 and 25 degrees C with the initial PO(4)(3-) concentration of 155 mg P/l. The FA samples treated in the same way described above can achieve 7.0 and 8.2 mg P/l phosphate removal for FA0.25 and FA700 respectively, corresponding to 45.2% and 52.9% removal. The activated materials performed higher phosphate removal over broader pH range compared with the raw ones. The influences of various factors, such as initial pH and initial phosphate concentration on the sorption capacity were also studied in batch equilibration technique. Solution pH significantly influenced the sorption. Each sample achieved the maximal removal of phosphate at pH 7.0. The amount of phosphate removal increased with the solute concentration. The Freundlich and Langmuir models were used to simulate the sorption equilibrium. The results indicate that the Langmuir model has a better correlation with the experimental data than the Freundlich model.
Assuntos
Carbono/química , Material Particulado/química , Fosfatos/química , Poluentes Químicos da Água/química , Purificação da Água/instrumentação , Purificação da Água/métodos , Adsorção , Cinza de Carvão , Relação Dose-Resposta a Droga , Concentração de Íons de Hidrogênio , Incineração , Resíduos Industriais , Metais Pesados , Microscopia Eletrônica de Varredura , Eliminação de Resíduos , Temperatura , Fatores de Tempo , ÁguaRESUMO
Dehydration-Responsive Element Binding ( DREB) transcription factors, specifically binding with dehydration reponsive element (DRE), activate a variety of stress-responsive genes in plants under abiotic stresses (dehydration, high salt and low temperature). Using PCR and homologous EST search, we isolated a DREB-like gene from Yinxin poplar (Populus alba x P. alba var. pyramidalis) named PaDREB2. Yeast One-hybrid experiment demonstrated that PaDREB2 protein could function as a DREB transcription factor activating target gene expression by specifically binding to DRE cis-element. To study the expression pattern of PaDREB2, RT-PCR was carried out. And the results showed that PaDREB2 is induced by low temperature, drought and high salt.
Assuntos
Proteínas de Plantas/metabolismo , Populus/genética , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , Clonagem Molecular , Temperatura Baixa , Secas , Etiquetas de Sequências Expressas , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/genética , Populus/metabolismo , Estresse Fisiológico , Fatores de Transcrição/genéticaRESUMO
The effects of colloidal and soluble organic substrates on nitrification in biofilms were investigated in biological aerated filters. The results showed that the hydrolysis process of colloidal organic matter took place rapidly, which could not be the limiting step of its oxidation in the biofilm. Some 70% organic matter oxidation was accomplished at the 40 cm at the bottom of the both filters, while the increase of organic matter in the influent resulted in the displacement of nitrification from the bottom to the upper part of filter bed. It was observed that the greater reduction on nitrification was caused by colloidal organic matter than by soluble organics at the same COD concentration.