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1.
Plant Pathol J ; 40(3): 329-335, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38835304

RESUMO

Phytophthora root and stem rot (PRR), caused by Phytophthora sojae, can occur at any growth stage under poorly drained and humid conditions. The expansion of soybean cultivation in South Korean paddy fields has increased the frequency of PRR outbreaks. This study aimed to identify four P. sojae isolates newly collected from domestic fields and evaluate race-specific resistance using the hypocotyl inoculation technique. The four isolates exhibited various pathotypes, with GJ3053 exhibiting the highest virulence complexity. Two isolates, GJ3053 and AD3617, were screened from 205 soybeans, and 182 and 190 genotypes (88.8 and 92.7%, respectively) were susceptible to each isolate. Among these accessions, five genotypes resistant to both isolates were selected. These promising genotypes are candidates for the development of resistant soybean cultivars that can effectively control PRR through gene stacking.

2.
ACS Appl Mater Interfaces ; 16(17): 21509-21521, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38642038

RESUMO

In this study, we synthesized levan shell hydrophobic silica nanoclusters encapsulating doxorubicin (L-HSi-Dox) and evaluated their potential as ultrasound-responsive drug delivery systems for cancer treatment. L-HSi-Dox nanoclusters were successfully fabricated by integrating a hydrophobic silica nanoparticle-doxorubicin complex as the core and an amphiphilic levan carbohydrate polymer as the shell by using an electrospray technique. Characterization analyses confirmed the stability, size, and composition of the nanoclusters. In particular, the nanoclusters exhibited a controlled release of Dox under aqueous conditions, demonstrating their potential as efficient drug carriers. The levanic groups of the nanoclusters enhanced the targeted delivery of Dox to specific cancer cells. Furthermore, the synergism between the nanoclusters and ultrasound effectively reduced cell viability and induced cell death, particularly in the GLUT5-overexpressing MDA-MB-231 cells. In a tumor xenograft mouse model, treatment with the nanoclusters and ultrasound significantly reduced the tumor volume and weight without affecting the body weight. Collectively, these results highlight the potential of the L-HSi-Dox nanoclusters and ultrasound as promising drug delivery systems with an enhanced therapeutic efficacy for biomedical applications.


Assuntos
Doxorrubicina , Frutanos , Doxorrubicina/química , Doxorrubicina/farmacologia , Humanos , Animais , Frutanos/química , Frutanos/farmacologia , Camundongos , Linhagem Celular Tumoral , Portadores de Fármacos/química , Nanopartículas/química , Sistemas de Liberação de Medicamentos , Ondas Ultrassônicas , Camundongos Nus , Feminino , Sobrevivência Celular/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Neoplasias/tratamento farmacológico , Neoplasias/diagnóstico por imagem , Neoplasias/patologia , Dióxido de Silício/química , Ensaios Antitumorais Modelo de Xenoenxerto
3.
J Microbiol Biotechnol ; 34(4): 838-845, 2024 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-38247212

RESUMO

Excessive alcohol consumption can have serious negative consequences on health, including addiction, liver damage, and other long-term effects. The causes of hangovers include dehydration, alcohol and alcohol metabolite toxicity, and nutrient deficiency due to absorption disorders. Additionally, alcohol consumption can slow reaction times, making it more difficult to rapidly respond to situations that require quick thinking. Exposure to a large amount of ethanol can also negatively affect a person's righting reflex and balance. In this study, we evaluated the potential of lactic acid bacteria (LAB) to alleviate alcohol-induced effects and behavioral responses. Two LAB strains isolated from kimchi, Levilactobacillus brevis WiKim0168 and Leuconostoc mesenteroides WiKim0172, were selected for their ethanol tolerance and potential to alleviate hangover symptoms. Enzyme activity assays for alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) were then conducted to evaluate the role of these bacteria in alcohol metabolism. Through in vitro and in vivo studies, these strains were assessed for their ability to reduce blood alcohol concentrations and protect against alcohol-induced liver damage. The results indicated that these LAB strains possess significant ethanol tolerance and elevate ADH and ALDH activities. LAB administration remarkably reduced blood alcohol levels in rats after excessive alcohol consumption. Moreover, the LAB strains showed hepatoprotective effects and enhanced behavioral outcomes, highlighting their potential as probiotics for counteracting the adverse effects of alcohol consumption. These findings support the development of functional foods incorporating LAB strains that can mediate behavioral improvements following alcohol intake.


Assuntos
Álcool Desidrogenase , Aldeído Oxirredutases , Etanol , Lactobacillales , Probióticos , Animais , Etanol/metabolismo , Álcool Desidrogenase/metabolismo , Ratos , Masculino , Probióticos/administração & dosagem , Lactobacillales/metabolismo , Concentração Alcoólica no Sangue , Fígado/metabolismo , Fígado/efeitos dos fármacos , Administração Oral , Leuconostoc mesenteroides , Aldeído Desidrogenase/metabolismo , Levilactobacillus brevis/metabolismo , Ratos Sprague-Dawley , Alimentos Fermentados/microbiologia
4.
Plants (Basel) ; 12(20)2023 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-37896053

RESUMO

This study aimed to discover the quantitative trait loci (QTL) associated with a high seed protein content in soybean and unravel the potential candidate genes. We developed two recombinant inbred line populations: YS and SI, by crossing Saedanbaek (high protein) with YS2035-B-91-1-B-1 (low protein) and Saedanbaek with Ilmi (low protein), respectively, and evaluated the protein content for three consecutive years. Using single-nucleotide polymorphism (SNP)-marker-based linkage maps, four QTLs were located on chromosomes 15, 18, and 20 with high logarithm of odds values (5.9-55.0), contributing 5.5-66.0% phenotypic variance. In all three experimental years, qPSD20-1 and qPSD20-2 were stable and identified in overlapping positions in the YS and SI populations, respectively. Additionally, novel QTLs were identified on chromosomes 15 and 18. Considering the allelic sequence variation between parental lines, 28 annotated genes related to soybean seed protein-including starch, lipid, and fatty acid biosynthesis-related genes-were identified within the QTL regions. These genes could potentially affect protein accumulation during seed development, as well as sucrose and oil metabolism. Overall, this study offers insights into the genetic mechanisms underlying a high soybean protein content. The identified potential candidate genes can aid marker-assisted selection for developing soybean lines with an increased protein content.

5.
BMC Ophthalmol ; 23(1): 278, 2023 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-37328801

RESUMO

BACKGROUND: Selective laser trabeculoplasty (SLT) is relatively safe and effective in lowering intraocular pressure (IOP). However, although rare, complications can occur after SLT. This report describes a patient with choroidal detachment due to hypotony following SLT without anterior chamber (AC) inflammation. CASE PRESENTATION: A 67-year-old man was referred for elevated IOP in his left eye with advanced glaucomatous visual field loss. He had previously been diagnosed with idiopathic uveitic glaucoma in the left eye, for which he underwent laser iridotomy, trabeculectomy, and cataract surgery. At the first visit, the IOP of his left eye measured by Goldmann tonometry was 28 mmHg despite maximally tolerated medical treatment. SLT was performed in his left eye, resulting in an IOP of 7 mmHg 7 days later. At 3 weeks post-procedure, the patient experienced ocular pain and decreased visual acuity in his left eye. Slit-lamp examination revealed deep anterior chamber depth and no inflammation reaction, but the IOP in his left eye was 4 mmHg, and both fundus and B-scan ultrasonography showed serous choroidal detachment. All anti-glaucoma agents were stopped, and the patient was started on treatment with oral prednisolone and cyclopentolate eye drops. Three weeks later, choroidal detachment had resolved and the IOP in his left eye had stabilized at 8 mmHg. Follow-up 3 months later showed that the IOP in his left eye remained stable. CONCLUSIONS: Choroidal detachment-related hypotony is a rare complication of SLT. This possible complication following SLT should be informed to the patients and considered when performing the procedure.


Assuntos
Efusões Coroides , Glaucoma , Terapia a Laser , Trabeculectomia , Masculino , Humanos , Idoso , Trabeculectomia/efeitos adversos , Trabeculectomia/métodos , Resultado do Tratamento , Glaucoma/cirurgia , Pressão Intraocular , Malha Trabecular , Terapia a Laser/efeitos adversos , Terapia a Laser/métodos , Efusões Coroides/cirurgia , Lasers
6.
Artigo em Inglês | MEDLINE | ID: mdl-37114142

RESUMO

To evaluate the antiobesity effects of yellow and black soybean, C57BL/6 mice were provided with a normal diet, high-fat diet, HFD-containing yellow soybean powder (YS), and black soybean powder (BS) for six weeks. Compared with the HFD group, both YS and BS decreased body weight by 30.1% and 37.2% and fat in tissue by 33.3% and 55.8%, respectively. Simultaneously, both soybeans significantly reduced the serum triglyceride and total cholesterol levels and regulated the lipogenic mRNA expressions of Pparγ, Acc, and Fas genes in the liver, supporting reduced body adiposity. Furthermore, BS significantly increased Pgc-1α and Ucp1 mRNA expression levels in epididymal adipose tissue, indicating thermogenesis is the key mechanism of BS. Taken together, our findings suggest that both soybeans prevent high-fat diet-induced obesity in mice by regulating lipid metabolism, and BS, in particular, has a greater antiobesity potential than YS.

7.
Foods ; 12(1)2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36613443

RESUMO

This study investigated the effects of high hydrostatic pressure (HHP) treatment on the physicochemical properties of rice flour according to its moisture levels in order to develop new materials for processed rice foods. The rice varieties used were the Samkwang variety (normal and hard type) and the Shingil variety (processing and soft type). The moisture content of the rice flour was adjusted to 35-55% and it was treated with the HHP treatment at 400-600 MPa. The water absorption capacity, solubility, and swelling power of the rice flour increased as the moisture levels and pressure increased. The 600 MPa enzymatic hydrolysis-treated rice flour showed similar results to the heat-treated rice flour. Scanning electron microscopy showed few cavities, resulting in a dense structure. X-ray diffraction confirmed that the 23° peak, which indicates the degree of gelatinization, decreased with increasing moisture levels and pressure. The HHP treatment of the rice flour changed its physicochemical properties according to the moisture levels and pressure applied. These results can provide important information on the development and production of various foods.

8.
Int J Mol Sci ; 23(4)2022 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-35216500

RESUMO

Pod-shattering causes a significant yield loss in many soybean cultivars. Shattering-tolerant cultivars provide the most effective approach to minimizing this loss. We developed molecular markers for pod-shattering and validated them in soybeans with diverse genetic backgrounds. The genes Glyma.16g141200, Glyma.16g141500, and Glyma.16g076600, identified in our previous study by quantitative trait locus (QTL) mapping and whole-genome resequencing, were selected for marker development. The whole-genome resequencing of three parental lines (one shattering-tolerant and two shattering-susceptible) identified single nucleotide polymorphism (SNP) and/or insertion/deletion (InDel) regions within or near the selected genes. Two SNPs and one InDel were converted to Kompetitive Allele-Specific PCR (KASP) and InDel markers, respectively. The accuracy of the markers was examined in the two recombinant inbred line populations used for the QTL mapping, as well as the 120 varieties and elite lines, through allelic discrimination and phenotyping by the oven-drying method. Both types of markers successfully discriminated the pod shattering-tolerant and shattering-susceptible genotypes. The prediction accuracy, which was as high as 90.9% for the RILs and was 100% for the varieties and elite lines, also supported the accuracy and usefulness of these markers. Thus, the markers can be used effectively for genetic and genomic studies and the marker-assisted selection for pod-shattering tolerance in soybean.


Assuntos
Marcadores Genéticos/genética , Glycine max/genética , Mutação INDEL/genética , Polimorfismo de Nucleotídeo Único/genética , Alelos , Mapeamento Cromossômico/métodos , Genes de Plantas/genética , Genoma de Planta/genética , Genótipo , Fenótipo , Locos de Características Quantitativas/genética
9.
Plants (Basel) ; 10(9)2021 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-34579299

RESUMO

Different physiological and genetic studies show that the variations in the accumulation of pigment-stimulating metabolites result in color differences in soybean seed coats. The objective of this study was to analyze the nutrient contents and antioxidant potential in black, brown, and green seed-coated soybeans. Significant variations in protein (38.9-43.3%), oil (13.9-20.4%), total sugar (63.5-97.0 mg/g seed), total anthocyanin (3826.0-21,856.0 µg/g seed coat), total isoflavone (709.5-3394.3 µg/g seed), lutein (1.9-14.8 µg/g), total polyphenol (123.0-385.8 mg gallic acid/100 g seed), total flavonoid (22.1-208.5 mg catechin/100 g seed), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid (ABTS; 275.0-818.8 mg Trolox/100 g seed), and 2,2-diphenyl-1-picrylhydrazyl (DPPH; 96.3-579.7 mg Trolox/100 g seed) were found among the soybean genotypes. Ilpumgeomjeong2 contained the lowest protein but the highest oil and total sugar. The lowest oil-containing Wonheug had the highest protein content. Socheong2 was rich in all four variables of antioxidants. Anthocyanins were detected only in black soybeans but not in brown and green soybeans. The variation in isoflavone content was up to 5-fold among the soybean genotypes. This study could be a valuable resource for the selection and improvement of soybean because an understanding of the nutrient content and antioxidant potentials is useful to develop effective strategies for improving the economic traits; for example, the major emphasis of soybean breeding for fatty acids is to enhance the oleic and linoleic acid contents and to decrease linolenic acid content.

10.
Plants (Basel) ; 10(9)2021 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-34579348

RESUMO

Improving drought stress tolerance of soybean could be an effective way to minimize the yield reduction in the drought prevailing regions. Identification of drought tolerance-related quantitative trait loci (QTLs) is useful to facilitate the development of stress-tolerant varieties. This study aimed to identify the QTLs for drought tolerance in soybean using a recombinant inbred line (RIL) population developed from the cross between a drought-tolerant 'PI416937' and a susceptible 'Cheonsang' cultivar. Phenotyping was done with a weighted drought coefficient derived from the vegetative and reproductive traits. The genetic map was constructed using 2648 polymorphic SNP markers that distributed on 20 chromosomes with a mean genetic distance of 1.36 cM between markers. A total of 10 QTLs with 3.52-4.7 logarithm of odds value accounting for up to 12.9% phenotypic variance were identified on seven chromosomes. Five chromosomes-2, 7, 10, 14, and 20-contained one QTL each, and chromosomes 1 and 19 harbored two and three QTLs, respectively. The chromosomal locations of seven QTLs overlapped or located close to the related QTLs and/or potential candidate genes reported earlier. The QTLs and closely linked markers could be utilized in maker-assisted selection to accelerate the breeding for drought tolerance in soybean.

11.
Nat Commun ; 12(1): 1395, 2021 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-33654088

RESUMO

On-chip glycan biosynthesis is an effective strategy for preparing useful complex glycan sources and for preparing glycan-involved applications simultaneously. However, current methods have some limitations when analyzing biosynthesized glycans and optimizing enzymatic reactions, which could result in undefined glycan structures on a surface, leading to unequal and unreliable results. In this work, a glycan chip is developed by introducing a pH-responsive i-motif DNA linker to control the immobilization and isolation of glycans on chip surfaces in a pH-dependent manner. On-chip enzymatic glycosylations are optimized for uniform biosynthesis of cancer-associated Globo H hexasaccharide and its related complex glycans through stepwise quantitative analyses of isolated products from the surface. Successful interaction analyses of the anti-Globo H antibody and MCF-7 breast cancer cells with on-chip biosynthesized Globo H-related glycans demonstrate the feasibility of the structure-switchable DNA linker-based glycan chip platform for on-chip complex glycan biosynthesis and glycan-involved applications.


Assuntos
DNA/metabolismo , Neoplasias/metabolismo , Polissacarídeos/biossíntese , Antígenos Glicosídicos Associados a Tumores/metabolismo , Toxina da Cólera/metabolismo , Gangliosídeo G(M1)/metabolismo , Glicosilação , Humanos , Concentração de Íons de Hidrogênio , Células MCF-7 , Análise de Sequência com Séries de Oligonucleotídeos , Polissacarídeos/química , Subunidades Proteicas/metabolismo
12.
Plants (Basel) ; 9(9)2020 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-32911865

RESUMO

Pod shattering is an important reproductive process in many wild species. However, pod shattering at the maturing stage can result in severe yield loss. The objectives of this study were to discover quantitative trait loci (QTLs) for pod shattering using two recombinant inbred line (RIL) populations derived from an elite cultivar having pod shattering tolerance, namely "Daewonkong", and to predict novel candidate QTL/genes involved in pod shattering based on their allele patterns. We found several QTLs with more than 10% phenotypic variance explained (PVE) on seven different chromosomes and found a novel candidate QTL on chromosome 16 (qPS-DS16-1) from the allele patterns in the QTL region. Out of the 41 annotated genes in the QTL region, six were found to contain SNP (single-nucleotide polymorphism)/indel variations in the coding sequence of the parents compared to the soybean reference genome. Among the six potential candidate genes, Glyma.16g076600, one of the genes with known function, showed a highly differential expression levels between the tolerant and susceptible parents in the growth stages R3 to R6. Further, Glyma.16g076600 is a homolog of AT4G19230 in Arabidopsis, whose function is related to abscisic acid catabolism. The results provide useful information to understand the genetic mechanism of pod shattering and could be used for improving the efficiency of marker-assisted selection for developing varieties of soybeans tolerant to pod shattering.

13.
ACS Appl Mater Interfaces ; 12(35): 38899-38905, 2020 Sep 02.
Artigo em Inglês | MEDLINE | ID: mdl-32805844

RESUMO

The work presented in this report demonstrates that amphiphilic polysaccharide-clasped self-assembly (Amp-SA) with nanometer size, encapsulating hydrophobic nanoparticles (NPs) can be generated via electrohydrodynamic spraying. It is observed that the formation of hydrophobic NP-encapsulated Amp-SA is dependent on the surface chemistry of NPs. The citrate-coated magnetic NPs (MNPs-Cit) were also prepared and compared. The hydrophobic magnetic NP-encapsulated Amp-SA (Amp-SA-M) exhibited around 2.7-2.8-fold higher values in r2 relaxivity than that of MNPs-Cit. In addition, the resulting Amp-SA-M achieved ∼17.2-fold higher values in r2/r1 ratios than MNPs-Cit. The enhanced performances in magnetic transverse (r2) relaxivity and r2/r1 ratio as well as the in vivo behavior of Amp-SA-M suggest the potential of Amp-SA-M as a promising MRI nanoprobe. This approach based on the nature-originated amphiphilic biopolymers may provide a novel insight into electrohydrodynamic techniques that have the ability to create various nanostructures, encapsulating high-quality hydrophobic nanomaterials for applications in diverse biotechnology.


Assuntos
Meios de Contraste/química , Nanopartículas de Magnetita/química , Polissacarídeos/química , Animais , Ácido Cítrico/química , Interações Hidrofóbicas e Hidrofílicas , Rim/diagnóstico por imagem , Imageamento por Ressonância Magnética , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Razão Sinal-Ruído
14.
Biomacromolecules ; 20(11): 4150-4157, 2019 11 11.
Artigo em Inglês | MEDLINE | ID: mdl-31545593

RESUMO

Polysaccharide-nanoparticle (NP) hybrid nanoclusters have great potential to revitalize diverse bioapplications; however, fabricating polysaccharide-based hybrid nanoclusters composed of high-quality NPs generated in the organic phase remains a challenge. Here, using calcium alginate as a polysaccharide/tetramethylammonium hydroxide (TMAOH) combination, we report a novel approach to the design of alginate-hydrophobic magnetic-plasmonic core-shell (MPCS) NP hybrid nanoclusters (A-MPCS HNCs). Furthermore, we observe the dependence of the formation of A-MPCS HNCs on the TMAOH concentration. The enhanced performance in both magnetic resonance r2 relaxivity and photoacoustic (PA) signals and the biocompatibility/bioactivity as well as the in vivo performance of A-MPCS HNCs shows them to be a promising magnetic resonance/photoacoustic dual-mode imaging agent. Our strategy could open doors to the use of other precious high-quality nanomaterials created in the organic phase via well-established synthetic chemistry in the design of alginate-hydrophobic nanomaterial hybrid nanoclusters, giving rise to novel and multifarious bioapplications.


Assuntos
Materiais Biocompatíveis/química , Nanopartículas/química , Nanoestruturas/química , Polissacarídeos/farmacologia , Alginatos/química , Alginatos/farmacologia , Ouro/química , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Imageamento por Ressonância Magnética , Espectroscopia de Ressonância Magnética , Técnicas Fotoacústicas , Polissacarídeos/química , Compostos de Amônio Quaternário/química , Compostos de Amônio Quaternário/farmacologia
15.
Chem Commun (Camb) ; 55(1): 71-74, 2018 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-30507991

RESUMO

A functional glycan chip combined with on-chip enzymatic glycosylation was developed to prepare complex glycan sources and to apply glycan-involved applications simultaneously. GM3 trisaccharide, GM2 tetrasaccharide, and GM1 pentasaccharide were successfully directly biosynthesized on lactose-immobilized surfaces through three consecutive glycosyltransferase reactions along with small amounts of enzymes and donors, without any additional processes. Biosynthesized GM1 pentasaccharide-related complex glycans were demonstrated to provide information on the substrate specificity of whole cholera toxin. Thus, the proposed on-chip glycan biosynthesis system can provide a new direction toward obtaining complex glycan sources and complex glycan-involved applications such as glycan-protein interaction analysis and glycan biomarker-based diagnosis.


Assuntos
Técnicas de Química Analítica/métodos , Oligossacarídeos/biossíntese , Sequência de Carboidratos , Toxina da Cólera/análise , Toxina da Cólera/metabolismo , Glicosilação , Glicosiltransferases/genética , Glicosiltransferases/metabolismo , Lactose/química , Lactose/metabolismo , Proteínas Recombinantes/metabolismo , Especificidade por Substrato
16.
ACS Appl Mater Interfaces ; 10(30): 25080-25089, 2018 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-29989402

RESUMO

Relaxivity tuning of nanomaterials with the intrinsic T1- T2 dual-contrast ability has great potential for MRI applications. Until now, the relaxivity tuning of T1 and T2 dual-modal MRI nanoprobes has been accomplished through the dopant, size, and morphology of the nanoprobes, leaving room for bioapplications. However, a surface engineering method for the relaxivity tuning was seldom reported. Here, we report the novel relaxivity tuning method based on the surface engineering of dual-mode T1- T2 MRI nanoprobes (DMNPs), along with protein interaction monitoring with the DMNPs as a potential biosensor application. Core nanoparticles (NPs) of europium-doped iron oxide (EuIO) are prepared by a thermal decomposition method. As surface materials, citrate (Cit), alendronate (Ale), and poly(maleic anhydride- alt-1-octadecene)/poly(ethylene glycol) (PP) are employed for the relaxivity tuning of the NPs based on surface engineering, resulting in EuIO-Cit, EuIO-Ale, and EuIO-PP, respectively. The key achievement of the current study is that the surface materials of the DMNP have significant impacts on the r1 and r2 relaxivities. The correlation between the hydrophobicity of the surface material and longitudinal relaxivity ( r1) of EuIO NPs presents an exponential decay feature. The r1 relaxivity of EuIO-Cit is 13.2-fold higher than that of EuIO-PP. EuIO can act as T1- T2 dual-modal (EuIO-Cit) or T2-dominated MRI contrast agents (EuIO-PP) depending on the surface engineering. The feasibility of using the resulting nanosystem as a sensor for environmental changes, such as albumin interaction, was also explored. The albumin interaction on the DMNP shows both T1 and T2 relaxation time changes as mutually confirmative information. The relaxivity tuning approach based on the surface engineering may provide an insightful strategy for bioapplications of DMNPs and give a fresh impetus for the development of novel stimuli-responsive MRI nanoplatforms with T1 and T2 dual-modality for various biomedical applications.

17.
Biosens Bioelectron ; 99: 289-295, 2018 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-28780344

RESUMO

Cholera toxin is a major virulent agent of Vibrio cholerae, and it can rapidly lead to severe dehydration, shock, causing death within hours without appropriate clinical treatments. In this study, we present a method wherein unique and short peptides that bind to cholera toxin subunit B (CTX-B) were selected through M13 phage display. Biopanning over recombinant CTX-B led to rapid screening of a unique peptide with an amino acid sequence of VQCRLGPPWCAK, and the phage-displayed peptides analyzed using ELISA, were found to show specific affinities towards CTX-B. To address the use of affinity peptides in development of the biosensor, sequences of newly selected peptides were modified and chemically synthesized to create a series of affinity peptides. Performance of the biosensor was studied using plasmonic-based optical techniques: localized surface plasmon resonance (LSPR) and surface-enhanced Raman scattering (SERS). The limit of detection (LOD) obtained by LSPR with 3σ-rule was 1.89ng/mL, while SERS had a LOD of 3.51pg/mL. In both cases, the sensitivity was much higher than the previously reported values, and our sensor system was specific towards actual CTX-B secreted from V. cholera, but not for CTX-AB5.


Assuntos
Técnicas Biossensoriais , Toxina da Cólera/isolamento & purificação , Cólera/diagnóstico , Vibrio cholerae O1/isolamento & purificação , Sequência de Aminoácidos/genética , Bacteriófago M13/genética , Cólera/microbiologia , Toxina da Cólera/toxicidade , Humanos , Peptídeos/química , Peptídeos/genética , Vibrio cholerae O1/patogenicidade
18.
Nanoscale ; 9(37): 13976-13982, 2017 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-28920122

RESUMO

We have demonstrated that the Verwey transition, which is highly sensitive to impurities, survives in anisotropic Gd-doped magnetite nanoparticles. Transmission electron microscopy analysis shows that the nanoparticles are uniformly distributed. X-ray photoelectron spectroscopy and EDS mapping analysis confirm Gd-doping on the nanoparticles. The Verwey transition of the Gd-doped magnetite nanoparticles is robust and the temperature dependence of the magnetic moment (zero field cooling and field cooling) shows the same behaviour as that of the Verwey transition in bulk magnetite, at a lower transition temperature (∼110 K). In addition, irregularly shaped nanoparticles do not show the Verwey transition whereas square-shaped nanoparticles show the transition. Mössbauer spectral analysis shows that the slope of the magnetic hyperfine field and the electric quadrupole splitting change at the same temperature, meaning that the Verwey transition occurs at ∼110 K. These results would provide new insights into understanding the Verwey transition in nano-sized materials.

19.
Biotechnol Prog ; 32(4): 848-54, 2016 07 08.
Artigo em Inglês | MEDLINE | ID: mdl-27071499

RESUMO

Organophosphorus hydrolase (OPH) from Flavobacterium species is a membrane-associated homodimeric metalloenzyme and has its own signal peptide in its N-terminus. We found that OPH was translocated into the periplasmic space when the original signal peptide-containing OPH was expressed in recombinant Escherichia coli even though its translocation efficiency was relatively low. To investigate the usability of this OPH signal peptide for periplasmic expression of heterologous proteins in an E. coli system, we employed green fluorescent protein (GFP) as a cytoplasmic folding reporter and alkaline phosphatase (ALP) as a periplasmic folding reporter. We found that the OPH signal peptide was able to use both twin-arginine translocation (Tat) and general secretory (Sec) machineries by switching translocation pathways according to the nature of target proteins in E. coli. These results might be due to the lack of Sec-avoidance sequence in the c-region and a moderate hydrophobicity of the OPH signal peptide. Interestingly, the OPH signal peptide considerably enhanced the translocation efficiencies for both GFP and ALP compared with commonly used TorA and PelB signal peptides that have Tat and Sec pathway dependences, respectively. Therefore, this OPH signal peptide could be successfully used in recombinant E. coli system for efficient periplasmic production of target protein regardless of the subcellular localization where functional folding of the protein occurs. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:848-854, 2016.


Assuntos
Arildialquilfosfatase/química , Escherichia coli/metabolismo , Flavobacterium/enzimologia , Peptídeos/metabolismo , Periplasma/metabolismo , Arildialquilfosfatase/metabolismo , Escherichia coli/citologia , Peptídeos/química , Periplasma/química
20.
BMC Biotechnol ; 16: 16, 2016 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-26879700

RESUMO

BACKGROUND: von Willebrand factor (VWF) is a key load bearing domain for mamalian cell adhesion by binding various macromolecular ligands in extracellular matrix such as, collagens, elastin, and glycosaminoglycans. Interestingly, vWF like domains are also commonly found in load bearing systems of marine organisms such as in underwater adhesive of mussel and sea star, and nacre of marine abalone, and play a critical load bearing function. Recently, Proximal Thread Matrix Protein1 (PTMP1) in mussel composed of two vWF type A like domains has characterized and it is known to bind both mussel collagens and mammalian collagens. RESULTS: Here, we cloned and mass produced a recombinant PTMP1 from E. coli system after switching all the minor codons to the major codons of E. coli. Recombinant PTMP1 has an ability to enhance mouse osteoblast cell adhesion, spreading, and cell proliferation. In addition, PTMP1 showed vWF-like properties as promoting collagen expression as well as binding to collagen type I, subsequently enhanced cell viability. Consequently, we found that recombinant PTMP1 acts as a vWF domain by mediating cell adhesion, spreading, proliferation, and formation of actin cytoskeleton. CONCLUSIONS: This study suggests that both mammalian cell adhesion and marine underwater adhesion exploits a strong vWF-collagen interaction for successful wet adhesion. In addition, vWF like domains containing proteins including PTMP1 have a great potential for tissue engineering and the development of biomedical adhesives as a component for extra-cellular matrix.


Assuntos
Bivalves/genética , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Animais , Linhagem Celular , Sobrevivência Celular , Colágeno , Escherichia coli/genética , Camundongos , Estrutura Terciária de Proteína/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Fator de von Willebrand/genética
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