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1.
Materials (Basel) ; 17(12)2024 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-38930332

RESUMO

Oxygen reduction reaction (ORR) and oxygen evolutionc reaction (OER) are important chemical reactions for a rechargeable lithium-oxygen battery (LOB). Recently, high-entropy alloys and oxides have attracted much attention because they showed good electrocatalytic performance for oxygen evolution reaction (OER) and/or oxygen reduction reaction (ORR). In this study, we aimed to synthesize and characterize CoSn(OH)6 and two types of high-entropy perovskite hydroxides, that is, (Co0.2Cu0.2Fe0.2Mn0.2Mg0.2)Sn(OH)6 (CCFMMSOH) and (Co0.2Cu0.2Fe0.2Mn0.2Ni0.2)Sn(OH)6 (CCFMNSOH). TEM observation and XRD measurements revealed that the high-entropy hydroxides CCFMMSOH and CCFMNSOH had cubic crystals with sides of approximately 150-200 nm and crystal structures similar to those of perovskite-type CSOH. LSV measurement results showed that the high-entropy hydroxides CCFMMSOH and CCFMNSOH showed bifunctional catalytic functions for the ORR and OER. CCFMNSOH showed better catalytic performance than CCFMMSOH.

2.
Materials (Basel) ; 17(2)2024 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-38255488

RESUMO

Lithium-air batteries (LABs) have a theoretically high energy density. However, LABs have some issues, such as low energy efficiency, short life cycle, and high overpotential in charge-discharge cycles. To solve these issues electrocatalytic materials were developed for oxygen reduction reaction (ORR) and oxygen evolution reaction (OER), which significantly affect battery performance. In this study, we aimed to synthesize electrocatalytic N-doped carbon-based composite materials with solution plasma (SP) using Co or Ni as electrodes from organic solvents containing cup-stacked carbon nanotubes (CSCNTs), iron (II) phthalocyanine (FePc), and N-nethyl-2-pyrrolidinone (NMP). The synthesized N-doped carbon-based composite materials were characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD), Raman spectroscopy, and X-ray photoelectron spectroscopy (XPS). TEM observation and XPS measurements revealed that the synthesized carbon materials contained elemental N, Fe, and electrode-derived Co or Ni, leading to the successful synthesis of N-doped carbon-based composite materials. The electrocatalytic activity for ORR of the synthesized carbon-based composite materials was also evaluated using electrochemical measurements. The electrochemical measurements demonstrated that the electrocatalytic performance for ORR of N-doped carbon-based composite material including Fe and Co showed superiority to that of N-doped carbon-based composite material including Fe and Ni. The difference in the electrocatalytic performance for ORR is discussed regarding the difference in the specific surface area and the presence ratio of chemical bonding species.

3.
RSC Adv ; 12(27): 17481-17489, 2022 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-35765431

RESUMO

A massive amount of animal biomass is generated daily from livestock farms, agriculture, and food industries, causing environmental and ecological problems. The conversion of animal biomass into value-added products has recently gained considerable interest in materials science research. Herein, horse manure (HM) was utilized as a precursor for synthesizing nitrogen-doped carbons (NCs) via hydrothermal ammonia treatment and the post pyrolysis process. The ammonia concentration varied between 0.5, 1.0, and 1.5 M in the hydrothermal process. From the comprehensive characterization results, horse manure-derived nitrogen-doped carbons (HMNCs) exhibited an amorphous phase and a hierarchical nanoporous structure. The specific surface area decreased from 170.1 to 66.6 m2 g-1 as the ammonia concentration increased due to micropore deterioration. The nitrogen content was 0.90 atom% even with no ammonia treatment, indicating self-nitrogen doping. With hydrothermal ammonia treatment, the nitrogen content slightly enhanced up to 1.54 atom%. The electrocatalytic activity for the oxygen reduction reaction (ORR) of HMNCs in an alkaline solution was found to be related to nitrogen doping content and porous structure. The ORR activity of HMNCs mainly proceeded via a combination of two- and four-electron pathways. Although the ORR activity of HMNCs was still not satisfactory and comparable to that of a commercial Pt/carbon catalyst, it showed better long-term durability. The results obtained in this work provide the potential utilization of HM as a precursor for ORR catalysts and other related applications.

4.
Biochem Soc Trans ; 50(3): 1069-1079, 2022 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-35621119

RESUMO

The ∼30 Mb genomes of the Plasmodium parasites that cause malaria each encode ∼5000 genes, but the functions of the majority remain unknown. This is due to a paucity of functional annotation from sequence homology, which is compounded by low genetic tractability compared with many model organisms. In recent years technical breakthroughs have made forward and reverse genome-scale screens in Plasmodium possible. Furthermore, the adaptation of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-Associated protein 9 (CRISPR/Cas9) technology has dramatically improved gene editing efficiency at the single gene level. Here, we review the arrival of genetic screens in malaria parasites to analyse parasite gene function at a genome-scale and their impact on understanding parasite biology. CRISPR/Cas9 screens, which have revolutionised human and model organism research, have not yet been implemented in malaria parasites due to the need for more complex CRISPR/Cas9 gene targeting vector libraries. We therefore introduce the reader to CRISPR-based screens in the related apicomplexan Toxoplasma gondii and discuss how these approaches could be adapted to develop CRISPR/Cas9 based genome-scale genetic screens in malaria parasites. Moreover, since more than half of Plasmodium genes are required for normal asexual blood-stage reproduction, and cannot be targeted using knockout methods, we discuss how CRISPR/Cas9 could be used to scale up conditional gene knockdown approaches to systematically assign function to essential genes.


Assuntos
Parasitos , Plasmodium , Toxoplasma , Animais , Sistemas CRISPR-Cas/genética , Edição de Genes/métodos , Humanos , Parasitos/genética , Plasmodium/genética , Toxoplasma/genética
5.
Materials (Basel) ; 15(9)2022 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-35591605

RESUMO

Li-air batteries have attracted considerable attention as rechargeable secondary batteries with a high theoretical energy density of 11,400 kWh/g. However, the commercial application of Li-air batteries is hindered by issues such as low energy efficiency and a short lifetime (cycle numbers). To overcome these issues, it is important to select appropriate cathode materials that facilitate high battery performance. Carbon materials are expected to be ideal materials for cathodes due to their high electrical conductivity and porosity. The physicochemical properties of carbon materials are known to affect the performance of Li-air batteries because the redox reaction of oxygen, which is an important reaction for determining the performance of Li-air batteries, occurs on the carbon materials. In this study, we evaluated the effect of the surface modification of carbon cathode materials on the charge-discharge performance of Li-air batteries using commercial Ketjenblack (KB) and KB subjected to vacuum ultraviolet (VUV) irradiation as cathodes. The surface wettability of KB changed from hydrophobic to hydrophilic as a result of the VUV irradiation. The ratio of COOH and OH groups on the KB surface increased after VUV irradiation. Raman spectra demonstrated that no structural change in the KB before and after VUV irradiation was observed. The charge and discharge capacities of a Li-air battery using VUV-irradiated KB as the cathode decreased compared to original KB, whereas the cycling performance of the Li-air battery improved considerably. The sizes and shapes of the discharge products formed on the cathodes changed considerably due to the VUV irradiation. The difference in the cycling performance of the Li-air battery was discussed from the viewpoint of the chemical properties of KB and VUV-irradiated KB.

6.
Materials (Basel) ; 15(5)2022 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-35268863

RESUMO

Fuel cells are expected to serve as next-generation energy conversion devices owing to their high energy density, high power, and long life performance. The oxygen reduction reaction (ORR) is important for determining the performance of fuel cells; therefore, using catalysts to promote the ORR is essential for realizing the practical applications of fuel cells. Herein, we propose Nb-incorporated TiO2 as a suitable alternative to conventional Pt-based catalysts, because Nb doping has been reported to improve the conductivity and electron transfer number of TiO2. In addition, Nb-incorporated TiO2 can induce the electrocatalytic activity for the ORR. In this paper, we report the synthesis method for Nb-incorporated TiO2 through a hydrothermal process with and without additional load pressures. The electrocatalytic activity of the synthesized samples for the ORR was also demonstrated. In this process, the samples obtained under various load pressures exceeding the saturated vapor pressure featured a high content of Nb and crystalline TiNb2O7, resulting in an ellipsoidal morphology. X-ray diffraction results also revealed that, on increasing the Nb doping amounts, the diffraction peak of the anatase TiO2 shifted to a lower angle and the full width at half maximum decreased. This implies that the Ti atom is exchanged with the Nb atom during this process, resulting in a decrease in TiO2 crystallinity. At a doping level of 10%, Nb-incorporated TiO2 exhibited the best electrocatalytic activity in terms of the oxygen reduction current (iORR) and onset potential for the ORR (EORR); this suggests that 10% Nb-doped samples have the potential for enhancing electrocatalytic activity.

7.
Parasitol Int ; 86: 102479, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34628068

RESUMO

Plasmodium, the causative agents of malaria, are obligate intracellular organisms. In humans, pathogenesis is caused by the blood stage parasite, which multiplies within erythrocytes, thus erythrocyte invasion is an essential developmental step. Merozoite form parasites released into the blood stream coordinately secrets a panel of proteins from the microneme secretory organelles for gliding motility, establishment of a tight junction with a target naive erythrocyte, and subsequent internalization. A protein identified in Toxoplasma gondii facilitates microneme fusion with the plasma membrane for exocytosis; namely, acylated pleckstrin homology domain-containing protein (APH). To obtain insight into the differential microneme discharge by malaria parasites, in this study we analyzed the consequences of APH deletion in the rodent malaria model, Plasmodium yoelii, using a DiCre-based inducible knockout method. We found that APH deletion resulted in a reduction in parasite asexual growth and erythrocyte invasion, with some parasites retaining the ability to invade and grow without APH. APH deletion impaired the secretion of microneme proteins, MTRAP and AMA1, and upon contact with erythrocytes the secretion of MTRAP, but not AMA1, was observed. APH-deleted merozoites were able to attach to and deform erythrocytes, consistent with the observed MTRAP secretion. Tight junctions were formed, but echinocytosis after merozoite internalization into erythrocytes was significantly reduced, consistent with the observed absence of AMA1 secretion. Together with our observation that APH largely colocalized with MTRAP, but less with AMA1, we propose that APH is directly involved in MTRAP secretion; whereas any role of APH in AMA1 secretion is indirect in Plasmodium.


Assuntos
Antígenos de Protozoários/genética , Deleção de Genes , Plasmodium yoelii/genética , Proteínas de Protozoários/genética , Acilação , Antígenos de Protozoários/metabolismo , Plasmodium yoelii/metabolismo , Proteínas de Protozoários/metabolismo
8.
Sci Rep ; 11(1): 16959, 2021 08 20.
Artigo em Inglês | MEDLINE | ID: mdl-34417510

RESUMO

Babesia parasite invades exclusively red blood cell (RBC) in mammalian host and induces alterations to host cell for survival. Despite the importance of Babesia in livestock industry and emerging cases in humans, their basic biology is hampered by lack of suitable biological tools. In this study, we aimed to develop a synchronization method for Babesia bovis which causes the most pathogenic form of bovine babesiosis. Initially, we used compound 2 (C2), a specific inhibitor of cyclic GMP-dependent protein kinase (PKG), and a derivative of C2, ML10. While both inhibitors were able to prevent B. bovis egress from RBC and increased percentage of binary forms, removal of inhibitors from culture did not result in a synchronized egress of parasites. Because using PKG inhibitors alone was not efficient to induce a synchronized culture, we isolated viable and invasive B. bovis merozoites and showed dynamics of merozoite invasion and development in RBCs. Using isolated merozoites we showed that BbVEAP, VESA1-export associated protein, is essential for parasite development in the RBC while has no significant role in invasion. Given the importance of invasion for the establishment of infection, this study paves the way for finding novel antigens to be used in control strategies against bovine babesiosis.


Assuntos
Babesia bovis/fisiologia , Merozoítos/fisiologia , Parasitos/fisiologia , Animais , Babesia bovis/efeitos dos fármacos , Proteínas Quinases Dependentes de GMP Cíclico/antagonistas & inibidores , Proteínas Quinases Dependentes de GMP Cíclico/metabolismo , Eritrócitos/efeitos dos fármacos , Eritrócitos/parasitologia , Cinética , Merozoítos/efeitos dos fármacos , Parasitos/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Imagem com Lapso de Tempo
9.
Parasitol Int ; 85: 102435, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34390881

RESUMO

Malaria remains a heavy global burden on human health, and it is important to understand the molecular and cellular biology of the parasite to find targets for drug and vaccine development. The mouse malaria model is an essential tool to characterize the function of identified molecules; however, robust technologies for targeted gene deletions are still poorly developed for the widely used rodent malaria parasite, Plasmodium yoelii. To overcome this problem, we established a DiCre-loxP inducible knockout (iKO) system in P. yoelii, which showed more than 80% excision efficacy of the target locus and more than 90% reduction of locus transcripts 24 h (one cell cycle) after RAP administration. Using this developed system, cAMP-dependent protein kinase (PKAc) was inducibly disrupted and the phenotypes of the resulting PKAc-iKO parasites were analyzed. We found that PKAc-iKO parasites showed severe growth and erythrocyte invasion defects. We also found that disruption of PKAc impaired the secretion of AMA1 in P. yoelii, in contrast to a report showing no role of PKAc in AMA1 secretion in P. falciparum. This discrepancy may be related to the difference in the timing of AMA1 distribution to the merozoite surface, which occurs just after egress for P. falciparum, but after several minutes for P. yoelii. Secretions of PyEBL, Py235, and RON2 were not affected by the disruption of PKAc in P. yoelii. PyRON2 was already secreted to the merozoite surface immediately after merozoite egress, which is inconsistent with the current model that RON2 is injected into the erythrocyte cytosol. Further investigations are required to understand the role of RON2 exposed on the merozoite surface.


Assuntos
Antígenos de Protozoários/biossíntese , Proteínas Quinases Dependentes de AMP Cíclico/genética , Proteínas de Membrana/biossíntese , Plasmodium yoelii/genética , Proteínas de Protozoários/genética , Animais , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Feminino , Merozoítos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos ICR , Plasmodium yoelii/enzimologia , Plasmodium yoelii/metabolismo , Proteínas de Protozoários/biossíntese , Proteínas de Protozoários/metabolismo
10.
Sci Rep ; 11(1): 9107, 2021 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-33907262

RESUMO

Theileria equi, an intraerythrocytic protozoan parasite, causes equine piroplasmosis, a disease which negatively impacts the global horse industry. Genetic manipulation is one of the research tools under development as a control method for protozoan parasites, but this technique needs to be established for T. equi. Herein, we report on the first development of a stable transgenic T. equi line expressing enhanced green fluorescent protein/blasticidin S deaminase (eGFP/BSD). To express the exogenous fusion gene in T. equi, regulatory regions of the elongation factor-1 alpha (ef-1α) gene were identified in T. equi. An eGFP/BSD-expression cassette containing the ef-1α gene promoter and terminator regions was constructed and integrated into the T. equi genome. On day 9 post-transfection, blasticidin-resistant T. equi emerged. In the clonal line of T. equi obtained by limiting dilution, integration of the eGFP/BSD-expression cassette was confirmed in the designated B-locus of the ef-1α gene via PCR and Southern blot analyses. Parasitaemia dynamics between the transgenic and parental T. equi lines were comparable in vitro. The eGFP/BSD-expressing transgenic T. equi and the methodology used to generate it offer new opportunities for better understanding of T. equi biology, with the add-on possibility of discovering effective control methods against equine piroplasmosis.


Assuntos
Aminoidrolases/genética , Proteínas de Fluorescência Verde/genética , Organismos Geneticamente Modificados , Theileria/genética , Regulação da Expressão Gênica , Nucleosídeos/farmacologia , Fator 1 de Elongação de Peptídeos/genética , Plasmídeos , Theileria/efeitos dos fármacos , Transfecção
11.
Sci Rep ; 11(1): 6261, 2021 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-33737609

RESUMO

Cathode materials are essential for enhancing electrocatalytic activity in energy-conversion devices. Carbon is one of the most suitable cathodic materials for Li-O2 batteries owing to its chemical and thermal stability. Carbon materials synthesized from tributyl borate (TBB) using a nonthermal solution plasma method were characterized using x-ray diffraction, Raman, field emission scanning electron microscopy (FE-SEM), transmission electron microscopy, and x-ray photoelectron spectroscopy and were evaluated as additive materials for cathodes in a Li-O2 battery. Two separate carbon materials were formed at the same time, a carbon dispersed in solution and a carbon precipitate at the bottom of the reactor, which had amorphous and graphite-like structures, respectively. The amorphous carbon contained boron and tungsten carbide, and the graphite-like carbon had more defects and electronic conductivity. The crystallinity and density of defects in the graphite-like carbon could be tuned by changing the SP operating frequency. The Li-O2 battery with the amorphous carbon containing boron and tungsten carbide was found to have a high capacity, while the one with the graphite-like carbon showed an affinity for the formation of Li2O2, which is the desired discharge product, and exhibited high cycling performance.

12.
mBio ; 11(1)2020 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-31911494

RESUMO

Erythrocyte-binding-like (EBL) proteins are known to play an important role in malaria parasite invasion of red blood cells (RBCs); however, any roles of EBL proteins in regulating host immune responses remain unknown. Here, we show that Plasmodium yoelii EBL (PyEBL) can shape disease severity by modulating the surface structure of infected RBCs (iRBCs) and host immune responses. We identified an amino acid substitution (a change of C to Y at position 741 [C741Y]) in the protein trafficking domain of PyEBL between isogenic P. yoelliinigeriensis strain N67 and N67C parasites that produce different disease phenotypes in C57BL/6 mice. Exchanges of the C741Y alleles altered parasite growth and host survival accordingly. The C741Y substitution also changed protein processing and trafficking in merozoites and in the cytoplasm of iRBCs, reduced PyEBL binding to band 3, increased phosphatidylserine (PS) surface exposure, and elevated the osmotic fragility of iRBCs, but it did not affect invasion of RBCs in vitro The modified iRBC surface triggered PS-CD36-mediated phagocytosis of iRBCs, host type I interferon (IFN-I) signaling, and T cell differentiation, leading to improved host survival. This study reveals a previously unknown role of PyEBL in regulating host-pathogen interaction and innate immune responses, which may be explored for developing disease control strategies.IMPORTANCE Malaria is a deadly parasitic disease that continues to afflict hundreds of millions of people every year. Infections with malaria parasites can be asymptomatic, with mild symptoms, or fatal, depending on a delicate balance of host immune responses. Malaria parasites enter host red blood cells (RBCs) through interactions between parasite ligands and host receptors, such as erythrocyte-binding-like (EBL) proteins and host Duffy antigen receptor for chemokines (DARC). Plasmodium yoelii EBL (PyEBL) is known to play a role in parasite invasion of RBCs. Here, we show that PyEBL also affects disease severity through modulation of host immune responses, particularly type I interferon (IFN-I) signaling. This discovery assigns a new function to PyEBL and provides a mechanism for developing disease control strategies.


Assuntos
Antígenos de Protozoários/imunologia , Eritrócitos/imunologia , Eritrócitos/parasitologia , Malária/imunologia , Malária/parasitologia , Proteínas de Membrana/metabolismo , Plasmodium yoelii/fisiologia , Proteínas de Protozoários/metabolismo , Alelos , Antígenos de Protozoários/metabolismo , Biomarcadores , Citocinas/metabolismo , Imunofluorescência , Interações Hospedeiro-Parasita , Imuno-Histoquímica , Malária/diagnóstico , Malária/metabolismo , Proteínas de Membrana/imunologia , Fragilidade Osmótica , Fagocitose/imunologia , Proteínas de Protozoários/genética , Proteínas de Protozoários/imunologia , Índice de Gravidade de Doença , Baço/imunologia , Baço/metabolismo , Baço/patologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo
13.
Parasitol Int ; 76: 102056, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31953169

RESUMO

Malaria parasites proliferate by repeated invasion of and multiplication within erythrocytes in the vertebrate host. Sexually committed intraerythrocytic parasites undergo sexual stage differentiation to become gametocytes. After ingestion by the mosquito, male and female gametocytes egress from erythrocytes and fertilize within the mosquito midgut. A complex signaling pathway likely responds to environmental events to trigger gametogenesis and regulate fertilization; however, such knowledge remains limited for malaria parasites. Several pseudokinases are highly transcribed at the gametocyte stage and are possible multi-functional regulators controlling critical steps of the life cycle. Here we characterized one pseudokinase, termed PypPK1, in Plasmodium yoelii that is highly expressed in schizonts and male gametocytes. Immunofluorescence assays for parasites expressing Myc-tagged PypPK1 confirmed that PypPK1 protein is expressed in schizonts and sexual stage parasites. Transgenic ΔpPK1 parasites, in which the PypPK1 gene locus was deleted by the CRISPR/Cas9 method, showed significant growth defect and reduced virulence in mice. In the blood stage, ΔpPK1 parasites were able to egress from erythrocytes similar to wild type parasites; however, erythrocyte invasion efficacy was significantly reduced. During sexual stage development, no clear changes were seen in male and female gametocytemias as well as gametocyte egress from erythrocytes; but, the number of exflagellation centers and oocysts were significantly reduced in ΔpPK1 parasites. Taken together, PypPK1 has an important role for both erythrocyte invasion and exflagellation center formation.


Assuntos
Eritrócitos/parasitologia , Plasmodium yoelii/enzimologia , Proteínas de Protozoários/genética , Animais , Feminino , Gametogênese , Estágios do Ciclo de Vida , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Plasmodium yoelii/patogenicidade , Proteínas de Protozoários/metabolismo , Esquizontes/enzimologia , Esquizontes/patogenicidade
14.
Sci Rep ; 9(1): 12704, 2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31481710

RESUMO

The metal-air battery is a form of renewable energy generation technology that produces energy electrochemically and can address energy concerns in the near future. However, state-of-the-art Pt electrocatalysts often suffer from agglomeration or detachment from carbon supports under prolonged operation, eventually limiting the long-term utilization of metal-air batteries. In this work, Pt nanoparticles were deposited on sulfur-doped nanocarbon to increase its stability. We first synthesized sulfur-doped (S-doped) and pristine carbon as support materials via a plasma process, and thereafter loaded platinum (Pt) nanoparticles onto the S-doped and pristine carbon matrix. From a sintering test at 600 °C, the Pt nanoparticles supported on pristine carbon increased from 2.4 to 5.2 nm; meanwhile, the average size of Pt NPs supported on S-doped carbon only increased from 2.2 to 2.51 nm. From the electrochemical analyses, the mass activity of Pt on pristine and S-doped carbon supports decreased by 25% and 10%, respectively, after 1500 cycles. The results proposed that the sulfide C-S-C bond provided a strong platinum-S-doped carbon support interaction between the support materials and the loaded Pt nanoparticles. Thus, S-doped carbon supports can serve as a stabilizer of Pt nanoparticles to enhance their durability in the application of metal-air batteries and other electrochemical devices.

15.
mSphere ; 4(3)2019 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-31189559

RESUMO

Babesia bovis, the most virulent causative agent of bovine babesiosis, is prevalent in tropical and subtropical regions of the world. Although the whole-genome sequence was released more than a decade ago, functional analysis of the genomics of this parasite is hampered by the limited breadth of genetic engineering tools. In this study, we implemented the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system for B. bovis and demonstrated its potential for genome editing. Cas9 and human dihydrofolate reductase (hDHFR) were simultaneously expressed by the B. boviselongation factor-1α bidirectional promoter, and a single guide RNA was expressed via the B. bovisU6 spliceosomal RNA promoter. Using a single plasmid construct, we were able to add an epitope tag to spherical body protein 3 (SBP3), introduce a point mutation into thioredoxin peroxidase 1 (tpx-1) to impair the function of the product, and replace the tpx-1 open reading frame with the other protein. Epitope tagging of SBP3 was efficient using this system, with a negligible number of remaining wild-type parasites and a pure transgenic population produced by allelic replacement of tpx-1 This advancement in genetic engineering tools for B. bovis will aid functional analysis of the genome and underpin characterization of candidate drug and vaccine targets.IMPORTANCEBabesia bovis is the most virulent cause of bovine babesiosis worldwide. The disease consequences are death, abortion, and economical loss due to reduced milk and meat production. Available vaccines are not effective, treatment options are limited, and emergence of drug and acaricide resistance has been reported from different regions. There is an urgent need to identify new drug and vaccine targets. Greater than half of the genes in B. bovis genome, including several expanded gene families which are unique for Babesia spp., have no predicted function. The available genetic engineering tools are based on conventional homologous recombination, which is time-consuming and inefficient. In this study, we adapted the CRISPR/Cas9 system as a robust genetic engineering tool for B. bovis This advancement will aid future functional studies of uncharacterized genes.


Assuntos
Babesia bovis/genética , Sistemas CRISPR-Cas , Edição de Genes , Deleção de Genes , Proteínas de Fluorescência Verde/genética , Humanos , Plasmídeos/genética , Mutação Puntual , Tetra-Hidrofolato Desidrogenase/genética
16.
Nanomaterials (Basel) ; 9(5)2019 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-31091814

RESUMO

Metal-air batteries are attracting increasing attention as a superior renewable energy conversion device due to their high performance and strong potential. However, the high cost and low stability of the current Pt catalyst is the main obstacle preventing wide industrial application. In this work, we applied a plasma process to fabricate aniline and a transition metals electrode (Fe, Co, Ni) as the carbon-nitrogen and the metal nanoparticle (NP) precursors, respectively, for selective metal/amino-N-doped carbon catalysts. All three as-synthesized catalysts exhibited dominant amino-N as the major C-N bonding state. In electrochemical testing, Co/amino-N-doped carbon showed positive E1/2 potential (0.83 V vs. Reversible Hydrogen Electrode (RHE)). In addition, the calculated electron transfer number (n) of Co/amino-N-doped carbon at 0.5 V vs. RHE was 3.81, which was only slightly less than that of commercial Pt/C (3.97). This superior performance of transition metal/amino-N-doped carbon promotes it as an economical oxygen reduction reaction (ORR) electrocatalyst to replace expensive Pt/C in metal-air batteries.

17.
Materials (Basel) ; 12(7)2019 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-30978976

RESUMO

Mg alloys are expected to be used in fields of the transportation industry because of their lightweight property, however, they show low corrosion resistance. To improve the corrosion resistance, preparation of the protective film on Mg alloys is essential. In this study, composite hydroxide films were prepared on three types of Mg alloys with different aluminum contents-that is, AZ31, AZ61, and AZ91D-by steam coating to investigate the relationship between the Mg-Al layered double hydroxide (LDH) content in the film and the Al content in the Mg alloys. Scanning electron microscopy (SEM) observation demonstrated that films were formed densely on all Mg alloy surfaces. X-ray diffraction (XRD) analyses revealed that all films prepared on AZ61 and AZ91D were composed of Mg(OH)2, AlOOH, and Mg-Al LDH, while the film containing Mg(OH)2 and Mg-Al LDH were formed only on AZ31. The Mg-Al LDH content in the film prepared on AZ61 was relatively higher than those prepared on AZ31 and AZ91D. The content of AlOOH in the film increased with an increase in the Al content in the Mg alloys. The film thickness changed depending on the treatment time and type of Mg alloy. Polarization curve measurements in 5 mass% NaCl solution demonstrated that the film prepared on the AZ61 showed complete passive behavior within the potential range of -1.0 to -0.64 V. In addition, immersion tests in 5 mass% NaCl aqueous solution for 480 h demonstrated that the film on the AZ61 had superior durability against 5 mass% NaCl aqueous solution. These results indicated that the film on the AZ61 had the most superior corrosion resistance among all samples. The results obtained in this study suggest that the LDH content in the film could be related to the corrosion resistance of the film.

18.
Materials (Basel) ; 11(9)2018 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-30205556

RESUMO

Corrosion resistant films with almost the same film thickness were prepared on the magnesium alloy AZ61 by steam coating at different vapor pressure and treatment times. The effect of the vapor pressure on the structures and the corrosion resistance of the films was investigated by using FE-SEM, SEM-EDX, GAXRD, and potentiodynamic polarization curve measurements in a 3.5 mass percentage NaCl aqueous solution. These studies clarified that the interlayers of Mg-Al Layered Double Hydroxide (LDHs) increased and its structure became non-uniform with an increase in the vapor pressure. The corrosion current density slightly increased with an increase in the vapor pressure during the treatment, but pitting corrosion occurred at both low and high vapor pressures. These results indicate that water molecules were pushed into an interlayer of Mg-Al LDHs by high vapor pressure. Consequently, the interlayer distance of Mg-Al LDH was widened and the cracks were generated in the anti-corrosive film. On the other hand, the Mg-Al LDH with an insufficiently large interlayer distance could not fill the cracks in the Mg(OH)2 crystallites and caused pitting corrosion when the vapor pressure was low.

19.
Parasitol Int ; 67(6): 706-714, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30025976

RESUMO

Plasmodium malaria parasites multiply within erythrocytes and possess a repertoire of proteins whose function is to recognize and invade these vertebrate host cells. One such protein involved in erythrocyte invasion is the micronemal protein, Erythrocyte Binding-Like (EBL), which has been studied as a potential target of vaccine development in Plasmodium vivax (PvDBP) and Plasmodium falciparum (EBA-175). In the rodent malaria parasite model Plasmodium yoelii, specific substitutions in the EBL regions responsible for intracellular trafficking (17XL parasite line) or receptor recognition (17X1.1pp. parasite line), paradoxically increase invasion ability and virulence rather than abolish EBL function. Attempts to disrupt the ebl gene locus in the 17XL and 17XNL lines were unsuccessful, suggesting EBL essentiality. To understand the mechanisms behind these potentially conflicting outcomes, we generated 17XL-based transfectants in which ebl expression is suppressed with anhydrotetracycline (ATc) and investigated merozoite behavior during erythrocyte invasion. In the absence of ATc, EBL was secreted to the merozoite surface, whereas following ATc administration parasitemia was negligible in vivo. Merozoites lacking EBL were unable to invade erythrocytes in vitro, indicating that EBL has a critical role for erythrocyte invasion. Quantitative time-lapse imaging revealed that with ATc administration a significant number of merozoites were detached from the erythrocyte after the erythrocyte deformation event and no echinocytosis was observed, indicating that EBL is required for merozoites to establish an irreversible connection with erythrocytes during invasion.


Assuntos
Antígenos de Protozoários/metabolismo , Eritrócitos/parasitologia , Malária/parasitologia , Parasitemia/parasitologia , Plasmodium yoelii/fisiologia , Proteínas de Protozoários/metabolismo , Receptores de Superfície Celular/metabolismo , Animais , Feminino , Técnicas de Silenciamento de Genes , Camundongos , Camundongos Endogâmicos ICR
20.
Colloids Surf B Biointerfaces ; 159: 655-661, 2017 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-28866413

RESUMO

Polymer brush, owing to its precisely controllable nanostructure, has great potential for surface modification in the biomedical field. In this study, we evaluated the bio-inertness of polymer brush, monomer monolayers, and polymer-coated surfaces based on their structures, to identify the most effective bio-inert modification. We focused on two well-known bio-inert materials, 2-methacryloyloxyethyl phosphorylcholine (MPC) and ethylene glycol (EG). The amount of adsorbed proteins on the surface was found to be dependent on the monomer unit density in the case of MPC, whereas this correlation was not observed in the case of EG. Cell adhesion was suppressed on the brush structure of both MPC and EG units, regardless of their density. The brush structure of MPC and EG units showed better anti-protein- and anti-cell-adhesion than monolayers and polymer-coated surfaces. Thus, the steric repulsion was not only important in EG units-based surface, but also in MPC-based surface. In addition, multiple polymer layers formed by MPC-based polymer coating also displayed similar properties.


Assuntos
Etilenoglicol/química , Metacrilatos/química , Fosforilcolina/análogos & derivados , Adesão Celular/fisiologia , Fosforilcolina/química , Polímeros/química , Proteínas/química , Propriedades de Superfície
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