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1.
Astrobiology ; 24(2): 138-150, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38393829

RESUMO

Most of the chemical and physical interactions of interest to the astrobiology community are influenced by the mineralogy of the systems under consideration. Often, this mineralogy occurs in sediment or sediment-like aqueous microenvironments in which the early minerals differ dramatically from the mature version that results from a long diagenesis, which are tied to complex interactions of pH, redox state, concentration, and temperature. This interconnectedness is difficult to reproduce in a laboratory setting yet is essential to understanding how the physical and chemical demands of living systems alter and are altered by their geological context. We present a facile means for producing precipitated mineral analogues within a microchannel and demonstrate its analytical efficacy through instrumental and modeling techniques. We show that amorphous, early-stage analogues of iron sulfide, iron carbonate, and iron phosphate can be formed at the boundary between flowing solutions, modeled on the microscale, and analyzed by standard instrumental techniques such as scanning electron microscopy/energy-dispersive spectroscopy, X-ray photoelectron spectroscopy, and Raman spectroscopy.


Assuntos
Compostos Ferrosos , Minerais , Fosfatos , Minerais/química , Carbonatos/análise , Ferro/química
2.
Phytopathology ; 113(10): 2006-2013, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37260102

RESUMO

Two infectious clones of turnip mosaic virus (TuMV), pKBC-1 and pKBC-8, with differential infectivity in Chinese cabbage (Brassica rapa subsp. pekinensis), were obtained. Both infected Nicotiana benthamiana systemically, inducing similar symptoms, whereas only virus KBC-8 infected Chinese cabbage systemically. To identify the determinants affecting infectivity on Chinese cabbage, chimeric clones were constructed by restriction fragment exchange between the parental clones and tested on several Chinese cabbage cultivars. Chimeric clones p1N8C and p8N1C demonstrated that the C-terminal portion of the polyprotein determines systemic infection of Chinese cabbage despite only three amino acid differences in this region, in the cylindrical inclusion (CI), viral protein genome-linked (VPg), and coat protein (CP). A second pair of hybrid constructs, pHindIII-1N8C and pHindIII-8N1C, failed to infect cultivars CR Victory and Jinseonnorang systemically, yet pHindIII-1N8C caused hypersensitive response-like lesions on inoculated leaves of these cultivars, and could systemically infect cultivars CR Chusarang and Jeongsang; this suggests that R genes effective against TuMV may exist in the first two cultivars but not the latter two. Constructs with single amino acid changes in both VPg (K2045E) and CP (Y3095H) failed to infect Chinese cabbage, implying that at least one of these two amino acid substitutions is essential for successful infection on Chinese cabbage. Successful infection by mutant KBC-8-CP-H and delayed infection with mutant HJY1-VPg-E following mutation or reversion suggested that VPg (2045K) is the residue required for infection of Chinese cabbage and involved in the interaction between VPg and eukaryotic initiation factor eIF(iso)4E, confirmed by yeast two-hybrid assay.


Assuntos
Brassica , Potyvirus , Aminoácidos/metabolismo , Doenças das Plantas , Potyvirus/genética
3.
Phytopathology ; 112(6): 1361-1372, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35113673

RESUMO

Three infectious clones of radish mosaic virus (RaMV) were generated from isolates collected in mainland Korea (RaMV-Gg) and Jeju Island (RaMV-Aa and RaMV-Bb). These isolates differed in sequences and pathogenicity. Examination of the wild-type isolates and reassortants between the genomic RNA1 and RNA2 of these three isolates revealed that severe symptoms were associated with RNA1 of isolates Aa or Gg causing systemic necrosis in Nicotiana benthamiana, or with RNA1 of isolate Bb for induction of veinal necrosis and severe mosaic symptoms in radish. Reverse transcription, followed by quantitative real-time PCR (Q-RT-PCR), results from infected N. benthamiana confirmed that viral RNA2 accumulation level was correlated to RaMV necrosis-inducing ability, and that the RNA2 accumulation level was mostly dependent on the origin of RNA1. However, in radish, Q-RT-PCR results showed more similar viral RNA2 accumulation levels regardless of the ability of the isolate to induce necrosis. Phylogenetic analysis of genomic RNAs sequence including previously characterized isolates from North America, Europe, and Asia suggest possible recombination within RNA1, while analysis of concatenated RNA1+RNA2 sequences indicates that reassortment of RNA1 and RNA2 has been more important in the evolution of RaMV isolates than recombination. Korean isolate Aa is a potential reassortant between isolates RaMV-J and RaMV-TW, while isolate Bb might have evolved from reassortment between isolates RaMV-CA and RaMV-J. The Korean isolates were shown to also be able to infect Chinese cabbage, raising concerns that RaMV may spread from radish fields to the Chinese cabbage crop in Korea, causing further economic losses.


Assuntos
Nicotiana , Raphanus , Células Clonais , Comovirus , Necrose , Filogenia , Doenças das Plantas , RNA Bacteriano , RNA Viral/genética
4.
Analyst ; 146(10): 3289-3298, 2021 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-33999058

RESUMO

An impedimetric biosensor is used to measure electrical impedance changes in the presence of biomolecules from sinusoidal input voltages. In this paper, we present a new portable impedance-based biosensor platform to improve the sensitivity of immunoassays with microparticles as a label. Using a 2 × 4 interdigitated electrode array with a 10/10 µm electrode/gap and a miniaturized impedance analyzer, we performed immunoassays with microparticles by integrating a microfluidic channel to evaluate signal enhancement. First, to understand the material dependency of microparticles on the sensor array, magnetic, silica, and polystyrene microparticles were tested. Among these microparticles, magnetic microparticles presented a high signal enhancement with relevant stability from the sensor array. With the magnetic microparticles, we demonstrate a series of immunoassays to detect human tumor necrosis factor (TNF-α) and compare the level of signal enhancement by measuring the limit of detection (LOD). With the microparticles, we achieved over ten times improvement of LOD from sandwich immunoassays. By incorporating with sample preparation and flow manipulation systems, this impedance sensor array can be utilized for digital diagnostics for a real sample-in answer-out system.


Assuntos
Técnicas Biossensoriais , Microfluídica , Impedância Elétrica , Humanos , Imunoensaio , Limite de Detecção
5.
Investig Clin Urol ; 61(6): 555-564, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-33135401

RESUMO

PURPOSE: Volumetry is used in polycystic kidney and liver diseases (PKLDs), including autosomal dominant polycystic kidney disease (ADPKD), to assess disease progression and drug efficiency. However, since no rapid and accurate method for volumetry has been developed, volumetry has not yet been established in clinical practice, hindering the development of therapies for PKLD. This study presents an artificial intelligence (AI)-based volumetry method for PKLD. MATERIALS AND METHODS: The performance of AI was first evaluated in comparison with ground-truth (GT). We trained a V-net-based convolutional neural network on 175 ADPKD computed tomography (CT) segmentations, which served as the GT and were agreed upon by 3 experts using images from 214 patients analyzed with volumetry. The dice similarity coefficient (DSC), interobserver correlation coefficient (ICC), and Bland-Altman plots of 39 GT and AI segmentations in the validation set were compared. Next, the performance of AI on the segmentation of 50 random CT images was compared with that of 11 PKLD specialists based on the resulting DSC and ICC. RESULTS: The DSC and ICC of the AI were 0.961 and 0.999729, respectively. The error rate was within 3% for approximately 95% of the CT scans (error<1%, 46.2%; 1%≤error<3%, 48.7%). Compared with the specialists, AI showed moderate performance. Furthermore, an outlier in our results confirmed that even PKLD specialists can make mistakes in volumetry. CONCLUSIONS: PKLD volumetry using AI was fast and accurate. AI performed comparably to human specialists, suggesting its use may be practical in clinical settings.


Assuntos
Cistos/patologia , Aprendizado Profundo , Hepatopatias/patologia , Doenças Renais Policísticas/patologia , Inteligência Artificial , Humanos
6.
Plant Pathol J ; 35(4): 381-387, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31481861

RESUMO

For several years, temperatures in the Korean peninsula have gradually increased due to climate change, resulting in a changing environment for growth of crops and vegetables. An associated consequence is that emerging species of insect vector have caused increased viral transmission. In Jeju Island, Korea, occurrences of viral disease have increased. Here, we report characterization of five newly collected turnip mosaic virus (TuMV) isolates named KBJ1, KBJ2, KBJ3, KBJ4 and KBJ5 from a survey on Jeju Island in 2017. Full-length cDNAs of each isolate were cloned into the pJY vector downstream of cauliflower mosaic virus 35S and bacteriophage T7 RNA polymerase promoters. Their fulllength sequences share 98.9-99.9% nucleotide sequence identity and were most closely related to previously reported Korean TuMV isolates. All isolates belonged to the BR group and infected both Chinese cabbage and radish. Four isolates induced very mild symptoms in Nicotiana benthamiana but KBJ5 induced a hypersensitive response. Symptom differences may result from three amino acid differences uniquely present in KBJ5; Gly(382)Asp, Ile(891)Val, and Lys(2522)Glu in P1, P3, and NIb, respectively.

8.
Arch Virol ; 164(6): 1553-1565, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30923966

RESUMO

Two isolates of Youcai mosaic virus (YoMV) were obtained, and their full-length genomic sequences were determined. Full-length infectious cDNA clones of each isolate were generated in which the viral sequence was under the control of dual T7 and 35S promoters for both in vitro transcript production and agro-infiltration. Comparison of the predicted amino acid sequences of the encoded proteins revealed only four differences between the isolates: three in the RNA-dependent RNA polymerase (RdRp) (V383I and M492I in the 125-kDa protein and T1245M in the 182-kDa protein); and one in the overlapping region of the movement protein (MP) and coat protein (CP) genes, affecting only the N-terminal domain of CP (CP M17T). One of the isolates caused severe symptoms in Nicotiana benthamiana plants, while the other caused only mild symptoms. In order to identify the amino acid residues associated with symptom severity, chimeric constructs were generated by combining parts of the two infectious YoMV clones, and the symptoms in infected plants were compared to those induced by the parental isolates. This allowed us to conclude that the M17T substitution in the N-terminal domain of CP was responsible for the difference in symptom severity. The M17T variation was found to be unique among characterized YoMV isolates. A difference in potential post-translational modification resulting from the presence of a predicted casein kinase II phosphorylation site only in the CP of isolate HK2 may be responsible for the symptom differences.


Assuntos
Nicotiana/virologia , Polimorfismo de Nucleotídeo Único , Tobamovirus/patogenicidade , Fatores de Virulência/genética , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/metabolismo , Doenças das Plantas , Processamento de Proteína Pós-Traducional , Fases de Leitura , Análise de Sequência de Proteína , Tobamovirus/genética , Proteínas Virais/genética , Proteínas Virais/metabolismo , Fatores de Virulência/metabolismo
9.
Phytopathology ; 109(5): 904-912, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30629482

RESUMO

Infectious clones were generated from 17 new Korean radish isolates of Turnip mosaic virus (TuMV). Phylogenetic analysis indicated that all new isolates, and three previously characterized Korean radish isolates, belong to the basal-BR group (indicating that the pathotype can infect both Brassica and Raphanus spp.). Pairwise analysis revealed genomic nucleotide and polyprotein amino acid identities of >87.9 and >95.7%, respectively. Five clones (HJY1, HJY2, KIH2, BE, and prior isolate R007) had lower sequence identities than other isolates and produced mild symptoms in Nicotiana benthamiana. These isolates formed three distinct sequence classes (HJY1/HJY2/R007, KIH2, and BE), and several differential amino acid residues (in P1, P3, 6K2, and VPg) were present only in mild isolates HJY1, HJY2, and R007. The remaining isolates all induced systemic necrosis in N. benthamiana. Four mild isolates formed a phylogenetic subclade separate from another subclade including all of the necrosis-inducing isolates plus mild isolate KIH2. Symptom severity in radish and Chinese cabbage genotypes was not correlated with pathogenicity in N. benthamiana; indeed, Chinese cabbage cultivar Norang was not infected by any isolate, whereas Chinese cabbage cultivar Chusarang was uniformly susceptible. Four isolates were unable to infect radish cultivar Iljin, but no specific amino acid residues were correlated with avirulence. These results may lead to the identification of new resistance genes against TuMV.


Assuntos
Brassica rapa/virologia , Nicotiana/virologia , Potyvirus/genética , Raphanus/virologia , Especificidade de Hospedeiro , Filogenia , Doenças das Plantas/virologia , Potyvirus/patogenicidade , Virulência
10.
Lab Chip ; 18(11): 1539-1551, 2018 05 29.
Artigo em Inglês | MEDLINE | ID: mdl-29736535

RESUMO

Animals are commonly used for pharmacokinetic studies which are the most frequent events tested during ocular drug development and preclinical evaluation. Inaccuracy, cost, and ethical criticism in these tests have created a need to construct an in vitro model for studying corneal constraints. In this work, a porous membrane embedded microfluidic platform is fabricated that separates a chip into an apical and basal side. After functionalizing the membrane surface with fibronectin, the membrane's mechanical and surface properties are measured to ensure correct modeling of in vivo characteristics. Immortalized human corneal epithelial cells are cultured on the membrane to create a microengineered corneal epithelium-on-a-chip (cornea chip) that is validated with experiments designed to test the barrier properties of the human corneal epithelium construct using model drugs. A pulsatile flow model is used that closely mimics the ocular precorneal constraints and is reasonable for permeability analysis that models in vivo conditions. This model can be used for preclinical evaluations of potential therapeutic drugs and to mimic the environment of the human cornea.


Assuntos
Epitélio Corneano , Dispositivos Lab-On-A-Chip , Técnicas Analíticas Microfluídicas/instrumentação , Modelos Biológicos , Soluções Oftálmicas/farmacocinética , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Epitélio Corneano/citologia , Epitélio Corneano/metabolismo , Desenho de Equipamento , Humanos , Membranas Artificiais , Técnicas Analíticas Microfluídicas/métodos , Permeabilidade
11.
Sci Rep ; 7(1): 1188, 2017 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-28446756

RESUMO

A chemically patterned microfluidic paper-based analytical device (C-µPAD) is developed to create fluidic networks by forming hydrophobic barriers using chemical vapor deposition (CVD) of trichlorosilane (TCS) on a chromatography paper. By controlling temperature, pattern size, and CVD duration, optimal conditions were determined by characterizing hydrophobicity, spreading patterns, and flow behavior on various sized fluidic patterns. With these optimal conditions, we demonstrated glucose assay, immunoassay, and heavy metal detection on well-spot C-µPAD and lateral flow C-µPAD. For these assays, standard curves showing correlation between target concentration and gray intensity were obtained to determine a limit of detection (LOD) of each assay. For the glucose assays on both well-spot C-µPAD and lateral flow C-µPAD, we achieved LOD of 13 mg/dL, which is equivalent to that of a commercial glucose sensor. Similar results were obtained from tumor necrosis factor alpha (TNFα) detection with 3 ng/mL of LOD. For Ni detection, a colorimetric agent was immobilized to obtain a stationary and uniform reaction by using thermal condensation coupling method. During the immobilization, we successfully functionalized amine for coupling the colorimetric agent on the C-µPAD and detected as low as 150 µg/L of Ni. These C-µPADs enable simple, rapid, and cost-effective bioassays and environmental monitoring, which provide practically relevant LODs with high expandability and adaptability.


Assuntos
Testes Diagnósticos de Rotina/instrumentação , Testes Diagnósticos de Rotina/métodos , Microfluídica/instrumentação , Microfluídica/métodos , Sistemas Automatizados de Assistência Junto ao Leito , Técnicas de Química Analítica , Imunoensaio/métodos , Papel
12.
Virus Genes ; 53(3): 434-445, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28176159

RESUMO

Two isolates of Pepper mild mottle virus (PMMoV) were selected from a nationwide survey of pepper fields in South Korea in 2014 and 2015, in which Cucumber mosaic virus was also detected; the two PMMoV isolates, Sangcheong 47 (S-47, KX399390) and Jeongsong 76 (J-76, KX399389), share ~99% nucleotide and amino acid identity and are closely related to Japanese and Chinese isolates at the nucleotide level. Amino acid sequence comparisons revealed 99.73, 99.81, 98.44, and 100% identity in the ORF1, ORF2, MP, and CP, respectively, between S-47 and J-76. In addition, we generated infectious clones of S-47 and J-76, and T7 promoter driven transcripts of each inoculated to Nicotiana benthamiana produced very severe symptoms, whereas only mild symptoms developed in Capsicum annuum. Gene silencing suppressor function of 126 kDa and cytoskeleton-connected plasmodesmata localization of movement protein of S-47 and J-76 showed no difference between isolates, whereas 126 kDa of J-76 clearly formed intracellular aggregates not observed with S-47 126 kDa protein. Differences between these isolates in 126/183 kDa-related functions including subcellular localization suggest that differential interactions with host proteins may affect symptom development in C. annuum.


Assuntos
Doenças das Plantas/virologia , Tobamovirus/isolamento & purificação , Tobamovirus/patogenicidade , Proteínas Virais/genética , Sequência de Aminoácidos , Substituição de Aminoácidos , Capsicum/virologia , Clonagem Molecular , Cucumovirus/genética , DNA Complementar/genética , Genoma Viral/genética , Filogenia , Regiões Promotoras Genéticas , Vírus de RNA/genética , República da Coreia , Nicotiana/virologia , Tobamovirus/genética
13.
Virus Genes ; 53(2): 286-299, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27913980

RESUMO

Seed-transmitted viruses have caused significant damage to watermelon crops in Korea in recent years, with cucumber green mottle mosaic virus (CGMMV) infection widespread as a result of infected seed lots. To determine the likely origin of CGMMV infection, we collected CGMMV isolates from watermelon and melon fields and generated full-length infectious cDNA clones. The full-length cDNAs were cloned into newly constructed binary vector pJY, which includes both the 35S and T7 promoters for versatile usage (agroinfiltration and in vitro RNA transcription) and a modified hepatitis delta virus ribozyme sequence to precisely cleave RNA transcripts at the 3' end of the tobamovirus genome. Three CGMMV isolates (OMpj, Wpj, and Mpj) were separately evaluated for infectivity in Nicotiana benthamiana, demonstrated by either Agroinfiltration or inoculation with in vitro RNA transcripts. CGMMV nucleotide identities to other tobamoviruses were calculated from pairwise alignments using DNAMAN. CGMMV identities were 49.89% to tobacco mosaic virus; 49.85% to pepper mild mottle virus; 50.47% to tomato mosaic virus; 60.9% to zucchini green mottle mosaic virus; and 60.96% to kyuri green mottle mosaic virus, confirming that CGMMV is a distinct species most similar to other cucurbit-infecting tobamoviruses. We further performed phylogenetic analysis to determine relationships of our new Korean CGMMV isolates to previously characterized isolates from Canada, China, India, Israel, Japan, Korea, Russia, Spain, and Taiwan available from NCBI. Analysis of CGMMV amino acid sequences showed three major clades, broadly typified as 'Russian,' 'Israeli,' and 'Asian' groups. All of our new Korean isolates fell within the 'Asian' clade. Neither the 128 nor 186 kDa RdRps of the three new isolates showed any detectable gene silencing suppressor function.


Assuntos
Cucumis sativus/virologia , Cucumovirus/genética , Filogenia , Doenças das Plantas/genética , Bacteriófago T7/genética , Citrullus/virologia , Cucumovirus/patogenicidade , Cucurbitaceae/virologia , DNA Complementar/genética , Genoma Viral , Doenças das Plantas/virologia , Regiões Promotoras Genéticas , Nicotiana/virologia , Vírus do Mosaico do Tabaco/genética , Tobamovirus/genética
14.
Virus Genes ; 52(4): 592-6, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27059238

RESUMO

In 2014, we performed a nationwide survey in Korean radish fields to investigate the distribution and variability of Turnip mosaic virus (TuMV). Brassica rapa ssp. pekinensis sap-inoculated with three isolates of TuMV from infected radish tissue showed different symptom severities, whereas symptoms in Raphanus sativus were similar for each isolate. The helper component-protease (HC-Pro) genes of each isolate were sequenced, and phylogenetic analysis showed that the three Korean isolates were clustered into the basal-BR group. The HC-Pro proteins of these isolates were tested for their RNA silencing suppressor (VSR) activity and subcellular localization in Nicotiana benthamiana. A VSR assay by co-agroinfiltration of HC-Pro with soluble-modified GFP (smGFP) showed that HC-Pro of isolate R007 and R041 showed stronger VSR activity than R065. The HC-Pros showed 98.25 % amino acid identity, and weak VSR isolate (R065) has a single variant residue in the C-terminal domain associated with protease activity and self-interaction compared to isolates with strong VSR activity. Formation of large subcellular aggregates of GFP:HC-Pro fusion proteins in N. benthamiana was only observed for HC-Pro from isolates with strong VSR activity, suggesting that R065 'weak' HC-Pro may have diminished self-association; substitution of the variant C-terminal residue largely reversed the HC-Pro aggregation and silencing suppressor characteristics. The lack of correlation between VSR efficiency and induction of systemic necrosis (SN) suggests that differences in viral accumulation due to HC-Pro are not responsible for SN.


Assuntos
Peptídeo Hidrolases/genética , RNA Viral/genética , Tymovirus/genética , Tymovirus/isolamento & purificação , Brassica napus/virologia , Cisteína Endopeptidases/genética , Coreia (Geográfico) , Filogenia , Doenças das Plantas/virologia , Interferência de RNA/fisiologia , Proteínas Virais/genética
15.
IEEE Trans Nanobioscience ; 14(8): 841-9, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26761951

RESUMO

Recent applications of PDMS nanocomposite materials and nanostructures have dramatically increased in biomedical fields due to optical, mechanical and electrical properties that are controllable by nanoengineering fabrication processes. These applications include biomedical imaging, biosensing, and cellular bioengineering studies using PDMS engineered structures with nanoparticles, nanopillars and functional nanoporous membranes. This article reviews the recent progress of PDMS nanocomposite materials and nanostructures and provides descriptions of various fabrication techniques. Together with these fabrication techniques, we discuss how these nanocomposite PDMS biomedical devices are revolutionizing biomedical science and engineering fields.


Assuntos
Bioengenharia/métodos , Nanocompostos , Nanoestruturas , Nanotecnologia/métodos , Linhagem Celular Tumoral , Humanos , Porosidade
16.
Am J Rhinol Allergy ; 28(2): 95-8, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24717942

RESUMO

BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) has various histological patterns and the immunologic characteristics differ between eastern and western patients. This study assessed the histological pattern of Korean NPs and determined whether Korean NP patterns have become westernized. METHODS: Three hundred forty-nine CRSwNP patients were enrolled in this study in 2001 (n = 107), 2006 (n = 111), and 2011 (n = 131). Histological type was categorized as eosinophilic, neutrophilic, seromucinous gland hyperplasia, and stromal atypia types. Medical records were reviewed for their atopic status, olfactory function, and computed tomography (CT) score. RESULTS: The eosinophilic type was significantly increased in 2011 (62.6%) when compared with 2001 (52.3%) and 2006 (47.7%). The incidence of atopic NPs was significantly higher in 2006 (18.9%) and 2011 (21.5%) than in 2001 (14.0%). Although atopic status and CT score were not significantly different among three NP types, anosmia was more common in eosinophilic NPs. CONCLUSION: The prevalence of NPs with atopy and eosinophilic NPs has continuously increased in Korea, indicative that the sinonasal disease pattern is becoming similar with western countries.


Assuntos
Eosinófilos/imunologia , Pólipos Nasais/diagnóstico , Seios Paranasais/patologia , Rinite/diagnóstico , Sinusite/diagnóstico , Adolescente , Adulto , Idoso , Doença Crônica , Progressão da Doença , Feminino , Humanos , Hiperplasia , Incidência , Coreia (Geográfico) , Masculino , Pessoa de Meia-Idade , Pólipos Nasais/imunologia , Pólipos Nasais/patologia , Seios Paranasais/diagnóstico por imagem , Prevalência , Rinite/imunologia , Rinite/patologia , Sinusite/imunologia , Sinusite/patologia , Olfato , Fatores de Tempo , Tomografia Computadorizada por Raios X , Adulto Jovem
17.
Arch Virol ; 159(6): 1373-83, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24378822

RESUMO

Soybean mosaic virus (SMV), a member of the family Potyviridae, is an important viral pathogen affecting soybean production in Korea. Variations in helper component proteinase (HC-Pro) sequences and the pathogenicity of SMV samples from seven Korean provinces were compared with those of previously characterized SMV isolates from China, Korea and the United States. Phylogenetic analysis separated 16 new Korean SMV isolates into two groups. Fourteen of the new Korean SMV samples belonged to group II and were very similar to U.S. strain SMV G7 and Chinese isolate C14. One isolate in group II, A297-13, differed at three amino acid positions (L54F, N286D, D369N) in the HC-Pro coding sequence from severe isolates and SMV 413, showed very weak silencing suppressor activity, and produced only mild symptoms in soybean. To test the role of each amino acid substitution in RNA silencing and viral RNA accumulation, a series of point mutations was constructed. Substitution of N for D at position 286 in HC-Pro of SMV A297-12 significantly reduced silencing suppression activity. When the mutant HC-Pro of A297-13 was introduced into an infectious clone of SMV 413, accumulation of viral RNA was reduced to less than 3 % of the level of SMV 413 containing HC-Pro of A297-12 at 10 days post-inoculation (dpi) but increased to 40 % of SMV 413(HC-Pro A297-12) at 40 dpi. At 50 dpi RNA accumulation of SMV 413(HC-Pro A297-13) was similar to that of SMV 413(HC-Pro A297-12). However, at 50 dpi, the D at position 286 of HC-Pro in SMV 413(HC-Pro A297-13) was found to have reverted to N. The results showed that 1) a naturally occurring mutation in HC-Pro significantly reduced silencing suppression activity and accumulation of transgene and viral RNAs, and 2) that there was strong selection for revision to wild type when the mutation was introduced into an infectious clone of SMV.


Assuntos
Cisteína Endopeptidases/genética , Glycine max/imunologia , Glycine max/virologia , Interações Hospedeiro-Patógeno , Potyvirus/genética , Potyvirus/imunologia , Interferência de RNA , Proteínas Virais/genética , Análise por Conglomerados , Cisteína Endopeptidases/metabolismo , Variação Genética , Coreia (Geográfico) , Dados de Sequência Molecular , Filogenia , Potyvirus/fisiologia , RNA Viral/genética , Análise de Sequência de DNA , Proteínas Virais/metabolismo
18.
Pharmacology ; 91(1-2): 39-47, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23154617

RESUMO

Bee venom (BV) has long been used as an oriental traditional medicine for the control of pain and inflammation. However, BV's anti-inflammatory mechanisms remain unclear. This study aimed to clarify the potential clinical efficacy of BV concerning the anti-inflammatory effect on nasal epithelial cell inflammation. Nasal polyp epithelial cells were obtained from patients. Cells were exposed to Alternaria alternata, Aspergillus nigra, Dermatophagoides pteronyssinus, Dermatophagoides farina and lipopolysaccharide with or without various concentrations of BV. Interleukin (IL)-6, IL-8, and granulocyte macrophage colony-stimulating factor were measured to determine the activation of epithelial cells. Nuclear factor-ĸB (NF-ĸB) and activator protein 1 expression and activity were determined with Western blot analysis and ELISA. Cytotoxicity of BV was measured using a CellTiter-96® aqueous cell proliferation assay kit. Cell survival was significantly decreased at BV concentrations exceeding 5 µg/ml. Fungi-induced cytokine production was more effectively inhibited by BV than house dust mite. Alternaria enhanced NF-ĸB expression, which was strongly inhibited by BV. BV appears to be relatively safe, and is of potential value for the treatment of airway inflammation and/or immunologic diseases.


Assuntos
Anti-Inflamatórios/farmacologia , Venenos de Abelha/farmacologia , Poluentes Atmosféricos , Alérgenos , Alternaria , Animais , Aspergillus niger , Células Cultivadas , Citocinas/metabolismo , Dermatophagoides farinae , Dermatophagoides pteronyssinus , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Humanos , NF-kappa B/metabolismo , Pólipos Nasais , Fator de Transcrição AP-1/metabolismo
19.
J Clin Biochem Nutr ; 50(2): 106-13, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22448090

RESUMO

Antioxidant supplementations are commonly used as an ergogenic aid for physical exercise despite its limited evidence. The study aimed to investigate the effects of a polyphenol mixture and vitamins on exercise endurance capacity. Seventy regularly exercising male participants were randomly assigned to receive oligomerized lychee fruit extract, a mixture of vitamin C (800 mg) and E (320 IU), or a placebo for 30 consecutive days. The study results showed that oligomerized lychee fruit extract significantly elevated the submaximal running time (p = 0.01). The adjusted mean change was 3.87 min (95% CI: 1.29, 6.46) for oligomerized lychee fruit extract, 1.33 (-1.23, 3.89) for the vitamins, and 1.60 (-1.36, 4.56) for the placebo (p = 0.33 in between groups). Oligomerized lychee fruit extract significantly increased the anaerobic threshold by 7.4% (1.8, 13.0). On the other hand, vitamins significantly attenuated VO(2)max by -3.11 ml/kg/m (-5.35, -0.87). Their effects on plasma free radical amount, however, were similar. Our results suggest that a polyphenol-containing supplement and typical antioxidants may have different mechanisms of action and that the endurance-promoting effect of oligomerized lychee fruit extract may not directly come from the scavenging of free radicals but may be attributed to other non-antioxidant properties of polyphenols, which requires further investigation.

20.
Lab Chip ; 11(18): 3106-12, 2011 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-21804972

RESUMO

A microfluidic device for solid-phase immunoassays based on microparticle labeling is developed using microvalve-control structures for automated sample processing. Programmable microvalve control in a multilayer structure provides automated sample delivery, adjustable hydrodynamic washing and compatibility with a wide range of substrates. Capture antibodies are derivatized on glass surfaces within the processor using an APTES patterning method, and magnetic microspheres conjugated with a secondary detection antibody are used as labels in a capture-sandwich format. In this microfluidic processor, washing force can be precisely controlled to remove the nonspecifically bound microparticles. Automated microfluidic immunoassays are demonstrated for mouse immunoglobulin (IgG) and human prostate specific antigen (PSA) with limits of detection of 1.8 and 3 pM, respectively. The sample processor architecture is easily parallelized for high-throughput analysis and easily interfaced with various assay substrates.


Assuntos
Anticorpos Imobilizados/química , Imunoensaio/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Animais , Anticorpos Imobilizados/metabolismo , Desenho de Equipamento , Humanos , Imunoensaio/métodos , Imunoglobulina G/análise , Imunoglobulina G/metabolismo , Camundongos , Pressão , Impressão , Antígeno Prostático Específico/análise , Antígeno Prostático Específico/metabolismo , Sensibilidade e Especificidade
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