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1.
J Oncol Pharm Pract ; 26(6): 1441-1451, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32615867

RESUMO

BACKGROUND: Tyrosine kinase inhibitors, commonly prescribed for non-small cell lung cancer, are efficacious but pose safety risks that can diminish quality of life. Quality measures to ensure appropriate utilization and monitoring of tyrosine kinase inhibitors are needed to reduce unnecessary healthcare costs and maximize therapeutic benefit. With integrated clinical and pharmaceutical care services, a health-system specialty pharmacy is the ideal setting for quality measures development. PURPOSE: The purpose of this initiative was to develop a tool to assess quality in the utilization of tyrosine kinase inhibitors in non-small cell lung cancer by engaging multiple key stakeholder perspectives to identify the fundamental elements of high-quality clinical and specialty pharmacy care. SUMMARY: Quality measures for the utilization of tyrosine kinase inhibitors in non-small cell lung cancer were developed by conducting package insert, guideline, and literature review followed by integrating feedback from key stakeholder groups through individual conversations and a structured assessment. The finalized guide assesses safety, efficacy, persistence, and patient-reported outcomes using defined clinical and specialty pharmacy measures. Most stakeholders agreed that the proposed measures were relevant as well as accurate and expressed support for implementation. CONCLUSIONS: The proposed tool provides a framework for specialty pharmacies to deliver quality care in the use of tyrosine kinase inhibitors for the treatment of non-small cell lung cancer. Further work is needed to implement and validate this tool.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Neoplasias Pulmonares/tratamento farmacológico , Inibidores de Proteínas Quinases/administração & dosagem , Custos de Cuidados de Saúde , Humanos , Assistência Farmacêutica/economia , Indicadores de Qualidade em Assistência à Saúde , Qualidade de Vida
2.
Int Ophthalmol ; 39(12): 2807-2813, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31134425

RESUMO

PURPOSE: To analyze biometric parameters and patterns of corneal astigmatism in cataract surgery patients using optical low-coherence interferometry in Northern United Arab Emirates (UAE). METHODS: Axial length (AL), anterior chamber depth (ACD), horizontal corneal diameter (white to white, WTW) and flat and steep keratometry (K1 and K2, respectively) were optically measured using optical low-coherence interferometry (Aladdin). Ocular datasets acquired between 2015 and 2018 were collected and analyzed. RESULTS: This study evaluated 238 eyes of 123 cataract patients with a mean age of 67.1 ± 9.4 years. Mean AL, ACD, WTW, K1, and K2 were 23.22 ± 0.99 mm, 3.05 ± 0.36 mm, 11.19 ± 0.46 mm, 43.72 ± 1.80 diopter (D), and 45.04 ± 1.71 D, respectively. Mean corneal astigmatism was 1.32 ± 0.97 D. Corneal astigmatism of 1.5 D or greater was found in 32.4%. With the rule, against the rule, and oblique astigmatism were found in 29.8%, 57.1%, and 13.0% of eyes, respectively. CONCLUSIONS: Results of this study provide normative data for cataract surgery as an informative reference in Northern UAE, the Middle East. This study showed higher corneal astigmatism in Northern UAE than those in other countries.


Assuntos
Astigmatismo/fisiopatologia , Extração de Catarata , Córnea , Complicações Pós-Operatórias/fisiopatologia , Idoso , Idoso de 80 Anos ou mais , Câmara Anterior/patologia , Astigmatismo/etiologia , Comprimento Axial do Olho/patologia , Biometria , Córnea/patologia , Córnea/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/etiologia , Emirados Árabes Unidos
3.
BMC Plant Biol ; 18(1): 118, 2018 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-29902970

RESUMO

BACKGROUND: Isoprene is a five-carbon chemical that is an important starting material for the synthesis of rubber, elastomers, and medicines. Although many plants produce huge amounts of isoprene, it is very difficult to obtain isoprene directly from plants because of its high volatility and increasing environmental regulations. Over the last decade, microorganisms have emerged as a promising alternative host for efficient and sustainable bioisoprene production. Isoprene synthase (IspS) has received much attention for the conversion of isoprene from dimethylallyl diphosphate (DMAPP). Herein, we isolated a highly expressible novel IspS gene from Metrosideros polymorpha (MpIspS), which was cloned and expressed in Escherichia coli, using a plant cDNA library and characterized its molecular and biochemical properties. RESULTS: The signal sequence deleted MpIspS was cloned and expressed in E. coli as a 65-kDa monomer. The maximal activity of the purified MpIspS was observed at pH 6.0 and 55 °C in the presence of 5 mM Mn2+. The Km, kcat, and kcat/Km for DMAPP as a substrate were 8.11 mM, 21 min- 1, and 2.59 mM- 1 min- 1, respectively. MpIspS was expressed along with the exogenous mevalonate pathway to produce isoprene in E. coli. The engineered cells produced isoprene concentrations of up to 23.3 mg/L using glycerol as the main carbon source. CONCLUSION: MpIspS was expressed in large amounts in E. coli, which led to increased enzymatic activity and resulted in isoprene production in vivo. These results demonstrate a new IspS enzyme that is useful as a key biocatalyst for bioisoprene production in engineered microbes.


Assuntos
Alquil e Aril Transferases/genética , Myrtaceae/enzimologia , Proteínas de Plantas/genética , Alquil e Aril Transferases/isolamento & purificação , Alquil e Aril Transferases/metabolismo , Butadienos/metabolismo , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Escherichia coli , Genes de Plantas/genética , Hemiterpenos/metabolismo , Microrganismos Geneticamente Modificados , Myrtaceae/genética , Filogenia , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , Alinhamento de Sequência
4.
Biochemistry ; 55(36): 5117-27, 2016 09 13.
Artigo em Inglês | MEDLINE | ID: mdl-27551953

RESUMO

Many bacteria, particularly pathogens, possess methionine sulfoxide reductase A (MsrA) and B (MsrB) as a fusion form (MsrAB). However, it is not clear why they possess a fusion MsrAB form rather than the separate enzymes that exist in most organisms. In this study, we performed biochemical and kinetic analyses of MsrAB from Treponema denticola (TdMsrAB), single-domain forms (TdMsrA and TdMsrB), and catalytic Cys mutants (TdMsrAB(C11S) and TdMsrAB(C285S)). We found that the catalytic efficiency of both MsrA and MsrB increased after fusion of the domains and that the linker region (iloop) that connects TdMsrA and TdMsrB is required for the higher catalytic efficiency of TdMsrAB. We also determined the crystal structure of TdMsrAB at 2.3 Å, showing that the iloop mainly interacts with TdMsrB via hydrogen bonds. Further kinetic analysis using the iloop mutants revealed that the iloop-TdMsrB interactions are critical to MsrB and MsrA activities. We also report the structure in which an oxidized form of dithiothreitol, an in vitro reductant for MsrA and MsrB, is present in the active site of TdMsrA. Collectively, the results of this study reveal an essential role of the iloop in maintaining the higher catalytic efficiency of the MsrAB fusion enzyme and provide a better understanding of why the MsrAB enzyme exists as a fused form.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/química , Biocatálise , Dicroísmo Circular , Proteínas Recombinantes de Fusão/química , Homologia de Sequência de Aminoácidos
5.
Biochem Biophys Res Commun ; 461(4): 648-52, 2015 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-25912135

RESUMO

Thioredoxin (Trx) is a major thiol-disulfide reductase that plays a role in many biological processes, including DNA replication and redox signaling. Although selenocysteine (Sec)-containing Trxs have been identified in certain bacteria, their enzymatic properties have not been characterized. In this study, we expressed a selenoprotein Trx from Treponema denticola, an oral spirochete, in Escherichia coli and characterized this selenoenzyme and its natural cysteine (Cys) homologue using E. coli Trx1 as a positive control. (75)Se metabolic labeling and mutation analyses showed that the SECIS (Sec insertion sequence) of T. denticola selenoprotein Trx is functional in the E. coli Sec insertion system with specific selenium incorporation into the Sec residue. The selenoprotein Trx exhibited approximately 10-fold higher catalytic activity than the Sec-to-Cys version and natural Cys homologue and E. coli Trx1, suggesting that Sec confers higher catalytic activity on this thiol-disulfide reductase. Kinetic analysis also showed that the selenoprotein Trx had a 30-fold higher Km than Cys-containing homologues, suggesting that this selenoenzyme is adapted to work efficiently with high concentrations of substrate. Collectively, the results of this study support the hypothesis that selenium utilization in oxidoreductase systems is primarily due to the catalytic advantage provided by the rare amino acid, Sec.


Assuntos
Selênio/química , Selenocisteína/química , Tiorredoxinas/química , Treponema denticola/enzimologia , Sítios de Ligação , Catálise , Ativação Enzimática , Ligação Proteica , Relação Estrutura-Atividade , Especificidade por Substrato
6.
Biochem Biophys Res Commun ; 457(4): 567-71, 2015 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-25600814

RESUMO

Glutaredoxin (Grx), a major redox regulator, can act as a reductant of methionine sulfoxide reductase A (MsrA). However, the biochemical mechanisms involved in MsrA activity regeneration by Grx remain largely unknown. In this study, we investigated the regeneration mechanism of 1-Cys type Clostridium oremlandii MsrA (cMsrA) lacking a resolving Cys residue in a Grx-dependent assay. Kinetic analysis showed that cMsrA could be reduced by both monothiol and dithiol Grxs as efficiently as by in vitro reductant dithiothreitol. Our data revealed that the catalytic Cys sulfenic acid intermediate is not glutathionylated in the presence of the substrate, and that Grx instead directly formed a complex with cMsrA. Mass spectrometry analysis identified a disulfide bond between the N-terminal catalytic Cys of the active site of Grx and the catalytic Cys of cMsrA. This mixed disulfide bond could be resolved by glutathione. Based on these findings, we propose a model for regeneration of 1-Cys type cMsrA by Grx that involves no glutathionylation on the catalytic Cys of cMsrA. This mechanism contrasts with that of the previously known 1-Cys type MsrB.


Assuntos
Clostridium/enzimologia , Glutarredoxinas/metabolismo , Metionina Sulfóxido Redutases/metabolismo , Sequência de Aminoácidos , Clostridium/química , Clostridium/metabolismo , Cisteína/análogos & derivados , Cisteína/química , Cisteína/metabolismo , Glutarredoxinas/química , Glutationa/metabolismo , Cinética , Metionina Sulfóxido Redutases/química , Dados de Sequência Molecular , Ácidos Sulfênicos/química , Ácidos Sulfênicos/metabolismo
7.
Arch Biochem Biophys ; 545: 1-8, 2014 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-24412203

RESUMO

Methionine sulfoxide reductase A (MsrA) reduces free and protein-based methionine-S-sulfoxide to methionine. Structures of 1-Cys MsrAs lacking a resolving Cys, which interacts with catalytic Cys, are unknown. In addition, no structural information on selenocysteine (Sec)-containing MsrA enzymes has been reported. In this work, we determined the crystal structures of 1-Cys type selenoprotein MsrA from Clostridium oremlandii at 1.6-1.8Å, including the reduced, oxidized (sulfenic acid), and substrate-bound forms. The overall structure of Clostridium MsrA, consisting of ten α-helices and six ß-strands, folds into a catalytic domain and a novel helical domain absent from other known MsrA structures. The helical domain, containing five helices, tightly interacts with the catalytic domain, and is likely critical for catalytic activity due to its association with organizing the active site. This helical domain is also conserved in several selenoprotein MsrAs. Our structural analysis reveals that the side chain length of Glu55 is critical for the proton donor function of this residue. Our structures also provide insights into the architecture of the 1-Cys MsrA active site and the roles of active site residues in substrate recognition and catalysis.


Assuntos
Clostridium/enzimologia , Cisteína/química , Metionina Sulfóxido Redutases/química , Sítios de Ligação , Domínio Catalítico , Clostridium/química , Cristalografia por Raios X , Cisteína/metabolismo , Metionina Sulfóxido Redutases/metabolismo , Modelos Moleculares , Oxirredução , Conformação Proteica , Estrutura Terciária de Proteína
8.
Nutrients ; 16(5)2024 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-38474817

RESUMO

Positional vertigo manifests as a spinning sensation triggered by changes in head position relative to gravity. Benign paroxysmal positional vertigo (BPPV) is an inner ear disorder characterized by recurrent episodes of positional vertigo. The connection between vitamin D insufficiency/deficiency and the onset and recurrence of BPPV is established. This study aims to assess vitamin D as a recurring factor in BPPV and the efficacy of vitamin D supplementation in preventing its recurrence. A comprehensive literature review on the relationship between vitamin D and BPPV recurrence was conducted, searching PubMed, Embase, Web of Science, and article reference lists for studies published from 2020 to 2023. A total of 79 articles were initially identified through the search, with 12 of them being utilized in the study. Recurrence rates for BPPV varied from 13.7% to 23% for studies with follow-up less than 1 year and 13.3% to 65% for studies with follow-up equal to or exceeding 2 years. Risk factors for BPPV recurrence include advanced age, female sex, hypertension, diabetes mellitus, hyperlipidemia, osteoporosis, and vitamin D deficiency. While earlier studies did not establish a link between low vitamin D levels and initial BPPV occurrence, they did associate recurrent episodes with low vitamin D levels. Recent research indicates that vitamin D supplementation in BPPV patients with deficiency or insufficiency decreases both the numbers of relapsing patients and relapses per patient. To validate these findings across diverse populations, further randomized controlled studies with larger cohorts and extended follow-up durations are essential.


Assuntos
Vertigem Posicional Paroxística Benigna , Deficiência de Vitamina D , Humanos , Feminino , Vertigem Posicional Paroxística Benigna/epidemiologia , Vertigem Posicional Paroxística Benigna/etiologia , Vertigem Posicional Paroxística Benigna/prevenção & controle , Vitamina D , Deficiência de Vitamina D/epidemiologia , Vitaminas , Recidiva , Suplementos Nutricionais
9.
J Clin Med ; 12(24)2023 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-38137794

RESUMO

The vestibular organ is involved in controlling blood pressure through vestibulosympathetic reflexes of the autonomic nervous system. This study aimed to investigate the effect of benign paroxysmal positional vertigo (BPPV) on blood pressure control by the autonomic nervous system by observing changes in blood pressure before and after BPPV treatment using the head-up tilt test (HUTT). A total of 278 patients who underwent the HUTT before and after treatment were included. The HUTT measured blood pressure repeatedly on the day of diagnosis and the day of complete recovery, and the results were analyzed using repeated measures analysis of variance. Regarding the difference in the systolic blood pressure of patients with BPPV, the blood pressure at 1, 2, and 3 min in the upright position after complete recovery was significantly lower than before treatment (p = 0.001, p = 0.001, and p = 0.012, respectively). Blood pressure at 1 and 2 min in the diastolic blood pressure of patients with BPPV in the upright position after complete recovery was significantly lower than before treatment (p = 0.001 and p = 0.034, respectively). This study shows that BPPV increases blood pressure during the initial response to standing in the HUTT.

10.
Mol Microbiol ; 79(5): 1194-203, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21210868

RESUMO

Several engineered selenocysteine (Sec)-containing glutaredoxins (Grxs) and their enzymatic properties have been reported, but natural selenoprotein Grxs have not been previously characterized. We expressed a bacterial selenoprotein Grx from Clostridium sp. (also known as Alkaliphilus oremlandii) OhILAs in Escherichia coli and characterized this selenoenzyme and its natural Cys homologues in Clostridium and E. coli. The selenoprotein Grx had a 200-fold higher activity than its Sec-to-Cys mutant form, suggesting that Sec is essential for catalysis by this thiol-disulfide oxidoreductase. Kinetic analysis also showed that the selenoprotein Grx had a 10-fold lower K(m) than Cys homologues. Interestingly, this selenoenzyme efficiently reduced a Clostridium selenoprotein methionine sulfoxide reductase A (MsrA), suggesting that it is the natural reductant for the protein that is not reducible by thioredoxin, a common reductant for Cys-containing MsrAs. We also found that the selenoprotein Grx could not efficiently reduce a Cys version of Clostridium MsrA, whereas natural Clostridium and E. coli Cys-containing Grxs, which efficiently reduce Cys-containing MsrAs, poorly acted on the selenoprotein MsrA. This specificity for MsrA reduction could explain why Sec is utilized in Clostridium Grx and more generally provides a novel example of the use of Sec in biological systems.


Assuntos
Proteínas de Bactérias/metabolismo , Clostridium/enzimologia , Glutarredoxinas/metabolismo , Metionina Sulfóxido Redutases/metabolismo , Selenoproteínas/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Clostridium/química , Clostridium/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Glutarredoxinas/química , Glutarredoxinas/genética , Cinética , Metionina Sulfóxido Redutases/genética , Dados de Sequência Molecular , Oxirredução , Engenharia de Proteínas , Selenocisteína/metabolismo , Selenoproteínas/genética , Alinhamento de Sequência
11.
Saudi J Ophthalmol ; 36(2): 189-194, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36211306

RESUMO

PURPOSE: The purpose of the study was to evaluate the agreement between measurements by optical low-coherence interferometry (OLCI, Aladdin) and those by Scheimpflug imaging combined with partial coherence interferometry (Scheimpflug-PCI, Pentacam AXL) in cataract patients. METHODS: This was a retrospective comparative study conducted in the United Arab Emirates. Axial length (AL), corneal power (keratometry, K), anterior chamber depth (ACD), and corneal astigmatism in patients with cataracts were measured with both devices. Difference and correlation were evaluated with paired t-test (p) and Pearson's correlation coefficient®, respectively. RESULTS: A total of 164 eyes of 95 patients were analyzed (164 eyes for K, 155 for ACD, and 112 for AL). The mean AL taken by OLCI was longer than that by Scheimpflug-PCI (23.25 mm vs. 23.23 mm, P ≤ 0.0001), showing an excellent correlation between the two (r = 0.9990). ACD measured by OLCI was 0.08 mm shallower than that by Scheimpflug-PCI (P = 0.0003, r = 0.7386). Corneal power measured by OLCI was lower than that by Scheimpflug-PCI (differences in mean K, flat K, and steep K were 0.05 diopters (D), 0.08 D, and 0.02 D, respectively), showing very strong correlations between the two devices (r = 0.9614, 0.9445, and 0.9535, respectively). Only flat K values measured with the two devices were significantly different (P = 0.0428). There were no statistically significant differences in the magnitude of astigmatism or J45 vector between the two devices (P = 0.1441 and P = 0.4147, respectively). However, J0 vector values were significantly different (P = 0.0087). CONCLUSION: Although OCLI and Scheimpflug-PCI showed strong correlations for measurements of AL, K, ACD, and corneal astigmatism in cataract patients, there were small but statistically significant differences in AL, ACD, flat K, and J0 vector. Thus, these two devices are not interchangeable for calculating intraocular lens power.

12.
Artigo em Inglês | MEDLINE | ID: mdl-34501609

RESUMO

Despite numerous studies on multiple socio-economic factors influencing urban PM2.5 pollution in China, only a few comparable studies have focused on developed countries. We analyzed the impact of three major socio-economic factors (i.e., income per capita, population density, and population size of a city) on PM2.5 concentrations for 254 cities from six developed countries. We used the Stochastic Impacts by Regression on Population, Affluence and Technology (STIRPAT) model with three separate data sets covering the period of 2001 to 2013. Each data set of 254 cities were further categorized into five subgroups of cities ranked by variable levels of income, density, and population. The results from the multivariate panel regression revealed a wide variation of coefficients. The most consistent results came from the six income coefficients, all of which met the statistical test of significance. All income coefficients except one carried negative signs, supporting the applicability of the environmental Kuznet curve. In contrast, the five density coefficients produced statistically significant positive signs, supporting the results from previous studies. However, we discovered an interesting U-shaped distribution of density coefficients across the six subgroups of cities, which may be unique to developed countries with urban pollution. The results from the population coefficients were not conclusive, which is similar to the results of previous studies. Implications from the results of this study for urban and national policy makers are discussed.


Assuntos
Poluentes Atmosféricos , Poluição do Ar , Poluentes Atmosféricos/análise , Poluição do Ar/análise , China , Cidades , Países Desenvolvidos , Monitoramento Ambiental , Material Particulado/análise , Densidade Demográfica
13.
Biochem Biophys Res Commun ; 395(3): 412-5, 2010 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-20382110

RESUMO

Free methionine-R-sulfoxide reductase (fRMsr) is a new type of methionine sulfoxide reductase that catalyzes the reduction of free methionine-R-sulfoxide to methionine. This enzyme cannot reduce oxidized methionine residues in proteins. While three Cys residues, Cys-91, Cys-101 and Cys-125, have been demonstrated to be involved in the catalysis by Saccharomyces cerevisiae fRMsr, their specific functions have not been fully established. In this work, we performed in vivo growth complementation experiments using S. cerevisiae cells lacking all three known methionine sulfoxide reductases. Cells containing a C125S construct, in which Cys-125 in fRMsr was replaced with Ser, did not grow in methionine sulfoxide medium, whereas cells containing C91S, C101S, or C91/101S constructs could grow in this medium. In addition, when assayed with thioredoxin and glutaredoxin reduction systems, the C125S form was inactive, whereas C91S and C101S had 1-2% and 9-10%, respectively, of the activity of the wild-type fRMsr. These data show that Cys-125 is the catalytic residue in fRMsr.


Assuntos
Cisteína/metabolismo , Metionina Sulfóxido Redutases/metabolismo , Saccharomyces cerevisiae/enzimologia , Domínio Catalítico/genética , Cisteína/genética , Teste de Complementação Genética , Metionina Sulfóxido Redutases/genética , Mutação , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento
14.
Clin Drug Investig ; 40(8): 727-735, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32474825

RESUMO

BACKGROUND AND OBJECTIVES: The vaccine adjuvant, AS04, present in bivalent human papilloma virus (bHPV) vaccines, induces greater local immune responses than the aluminum adjuvant in quadrivalent HPV (qHPV) vaccines. These distinctions might also result in disparities in immunogenicity and responsiveness, possibly contributing to differences in adverse events (AEs) between these vaccines. Here we comparatively analyzed AEs between qHPV and bHPV vaccines based on 10-year real-world AE data. METHODS: We evaluated the Korea Adverse Events Reporting System (KAERS), a nationwide drug database including vaccines from January 2007 to December 2016, analyzing AEs reported for qHPV and bHPV vaccines. Vaccine-AE pairs were generated, and the characteristics of all reported AEs were analyzed. Signals were derived using the disproportionality method of signal detection algorithms (reporting odds ratios and information component). RESULTS: Of the total 2566 HPV vaccine-associated AE reports, 2686 and 1994 were vaccine-AE pairs for qHPV and bHPV, respectively. Application site disorders were the most frequent AEs for both vaccines but were more frequently reported with the bHPV vaccine. The characteristics of non-application site AEs between the two vaccines were generally similar, but systemic AEs such as fever and fatigue were more common with the bHPV vaccine. Tremor, rash, eye pain, myopathy and circulatory failure were identified as signals in both qHPV and bHPV vaccines. CONCLUSIONS: Both application site disorders and systemic AEs were somewhat more frequent with the bHPV vaccine than with the qHPV vaccine. This might be caused by an immune response induced by adjuvants contained within the vaccines.


Assuntos
Aprovação de Drogas , Infecções por Papillomavirus/prevenção & controle , Vacinas contra Papillomavirus/uso terapêutico , Vigilância de Produtos Comercializados , Adolescente , Adulto , Algoritmos , Feminino , Humanos , Vacinas contra Papillomavirus/efeitos adversos , Vacinas contra Papillomavirus/imunologia , República da Coreia
15.
J Vet Sci ; 21(1): e4, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31940683

RESUMO

Fast and accurate detection of viral RNA pathogens is important in apiculture. A polymerase chain reaction (PCR)-based detection method has been developed, which is simple, specific, and sensitive. In this study, we rapidly (in 1 min) synthesized cDNA from the RNA of deformed wing virus (DWV)-infected bees (Apis mellifera), and then, within 10 min, amplified the target cDNA by ultra-rapid qPCR. The PCR products were hybridized to a DNA-chip for confirmation of target gene specificity. The results of this study suggest that our method might be a useful tool for detecting DWV, as well as for the diagnosis of RNA virus-mediated diseases on-site.


Assuntos
Abelhas/virologia , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , Vírus de RNA/isolamento & purificação , RNA Viral/análise , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Criação de Abelhas/métodos , DNA Complementar/análise , DNA Viral/análise , Vírus de RNA/genética
16.
PLoS One ; 12(1): e0170398, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28099480

RESUMO

Cellulose-binding domains (CBDs) are protein domains with cellulose-binding activity, and some act as leaders in the localization of cellulosomal scaffoldin proteins to the hydrophobic surface of crystalline cellulose. In this study, we found that a CBD fusion enhanced and improved soluble ß-glucuronidase (GusA) enzyme properties through the formation of an artificially oligomeric state. First, a soluble CBD fused to the C-terminus of GusA (GusA-CBD) was obtained and characterized. Interestingly, the soluble GusA-CBD showed maximum activity at higher temperatures (65°C) and more acidic pH values (pH 6.0) than free GusA did (60°C and pH 7.5). Moreover, the GusA-CBD enzyme showed higher thermal and pH stabilities than the free GusA enzyme did. Additionally, GusA-CBD showed higher enzymatic activity in the presence of methanol than free GusA did. Evaluation of the protease accessibility of both enzymes revealed that GusA-CBD retained 100% of its activity after 1 h incubation in 0.5 mg/ml protease K, while free GusA completely lost its activity. Simple fusion of CBD as a single domain may be useful for tunable enzyme states to improve enzyme stability in industrial applications.


Assuntos
Escherichia coli/metabolismo , Glucuronidase/metabolismo , Domínios Proteicos/fisiologia , Proteínas Recombinantes de Fusão/metabolismo , Sítios de Ligação , Celulose/metabolismo , Endopeptidase K/metabolismo , Estabilidade Enzimática , Escherichia coli/genética , Glucuronidase/genética , Temperatura Alta , Metanol/química , Ligação Proteica/fisiologia , Proteínas Recombinantes de Fusão/genética
17.
Yonsei Med J ; 46(2): 260-7, 2005 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-15861500

RESUMO

Endothelial progenitor cells (EPCs) have been reported to possess the capacity to colonize vascular grafts and hold promise for therapeutic neovascularization. However, limited quantities of EPCs have been the major factor impeding effective research on vasculoangiogenesis. In this study, cytokine and culture conditions necessary for the provision of large quantities of endothelial cells (ECs) were investigated. Cord blood was collected from 18 normal full-term deliveries and CD34+ cells were isolated by MACS system (Miltenyi Biotech, Bergish-Gladbach, Germany). To evaluate the effect of cytokines, CD34+ cells were cultured with various cytokine combinations, such as stem cell factor (SCF), flt3-ligand (FL), and thrombopoietin (TPO) with vascular endothelial growth factor (VEGF), interleukin-1 beta , fibroblast growth factor-basic (FGF-b) as basic cytokines. The quantities of non-adherent and adherent cells were the greatest with SCF, FL and TPO. The addition of TPO to all other cytokines significantly increased the number of non-adherent and adherent cells (p< 0.05, Wilcoxon rank sum test). After four weeks of culture, adherent cells expressed endothelial specific markers such as KDR, CD31 and CD62E. Typical morphology of ECs was observed during culture, such as cord-like structure and cobblestone appearance, suggesting that the adherent cells were consistent with ECs. In this study, the experimental conditions that optimize the production of ECs for therapeutic neovascularization were described. And it was possibly suggested that TPO plays a major role in differentiation from EPCs to ECs.


Assuntos
Separação Celular , Células Endoteliais , Sangue Fetal/citologia , Células-Tronco/imunologia , Antígenos CD34/análise , Diferenciação Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Células Cultivadas , Citocinas/farmacologia , Células Endoteliais/imunologia , Feto , Citometria de Fluxo , Humanos , Trombopoetina/farmacologia
18.
Clin Exp Emerg Med ; 2(3): 155-161, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27752590

RESUMO

OBJECTIVE: To investigate the association of red cell distribution width (RDW) with 30-day mortality in elderly patients with severe sepsis and septic shock. METHODS: Patients were recruited from a single tertiary emergency department. Patients with age over 65 years were selected. The main outcome was 30-day mortality. Potential confounders as Acute Physiologic and Chronic Health Evaluation (APACHE) II score and Sequential Organ Failure Assessment (SOFA) score along with initial vital signs were collected. Multivariate Cox proportional hazards analysis was performed to identify independent predictors of 30-day mortality. The discriminative ability of RDW for 30-day mortality was evaluated using receiver operating characteristic curve analysis. RESULTS: Overall, 458 patients were included. Univariate analysis showed that patients' survival was significantly associated with sites of infection, comorbidities, and severity scores. In the multivariate Cox proportional hazard model, the RDW was an independent predictor of 30-day mortality (hazards ratio, 1.10; 95% confidence interval, 1.04 to 1.17; P<0.001). CONCLUSION: In this study, initial RDW values were significantly associated with 30-day mortality in older patients hospitalized with severe sepsis and septic shock.

19.
IEEE Trans Inf Technol Biomed ; 16(4): 572-85, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22434818

RESUMO

Facing the increasing demands and challenges in the area of chronic disease care, various studies on the healthcare system which can, whenever and wherever, extract and process patient data have been conducted. Chronic diseases are the long-term diseases and require the processes of the real-time monitoring, multidimensional quantitative analysis, and the classification of patients' diagnostic information. A healthcare system for chronic diseases is characterized as an at-hospital and at-home service according to a targeted environment. Both services basically aim to provide patients with accurate diagnoses of disease by monitoring a variety of physical states with a number of monitoring methods, but there are differences between home and hospital environments, and the different characteristics should be considered in order to provide more accurate diagnoses for patients, especially, patients having chronic diseases. In this paper, we propose a patient status classification method for effectively identifying and classifying chronic diseases and show the validity of the proposed method. Furthermore, we present a new healthcare system architecture that integrates the at-home and at-hospital environment and discuss the applicability of the architecture using practical target services.


Assuntos
Doença Crônica/terapia , Prestação Integrada de Cuidados de Saúde/métodos , Serviços Hospitalares de Assistência Domiciliar , Informática Médica , Assistência Centrada no Paciente/métodos , Eletrocardiografia , Humanos , Síndrome Metabólica/terapia , Modelos Teóricos , Monitorização Fisiológica , Obesidade/terapia , Processamento de Sinais Assistido por Computador
20.
N Biotechnol ; 27(4): 435-9, 2010 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-20085831

RESUMO

This study investigated the stability of meoru (wild vine grape) anthocyanins in the aqueous solution under singlet oxygen. Freeze-dried meoru (1 kg) contained 179.98 mg anthocyanins including delphinidin-3-glucoside, malvidin-3,5-diglucoside, cyanidin-3,5-diglucoside, malvidin-3-glucoside, and cyanidin-3-glucoside. Malvidin-3,5-diglucoside and cyanidin-3-glucoside were the meoru anthocyanins at the highest and the lowest concentration, respectively. Little decrease in total anthocyanins in the aqueous solution was observed in the dark with or without riboflavin, or with light without riboflavin. Singlet oxygen degraded the meoru anthocyanins in the aqueous solution, which suggested chemical quenching of singlet oxygen by the anthocyanins. Degradation of the meoru anthocyanins was structure-dependent; diglucoside anthocyanins were more stable than monoglucoside. And malvidin glucoside was more stable than delphinidin or cyanidin glucoside, which suggested the number of hydroxy groups in the structure was partly related with the anthocyanin stability under singlet oxygen. This is the first report on anthocyanins stability affected by its structure under singlet oxygen.


Assuntos
Antocianinas/química , Fotoquímica/métodos , Riboflavina/química , Oxigênio Singlete/farmacologia , Vitis/química , Cromatografia Líquida de Alta Pressão , Luz , Soluções , Vitis/efeitos dos fármacos , Vitis/efeitos da radiação
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