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This study was to investigate the effects of different phytogenic feed additives (PFA) dosage levels in growing- finishing pigs stressed by high stocking density. A total of 72 mix sexed 12 weeks growing pigs ([Landrace × Yorkshire] × Duroc) with initial body weight (BW) of 49.28 ± 4.58 kg were used for 8 weeks. There were 3 replicate pens in each treatment group, with 3 pigs per pen. The dietary treatment groups consisted of basal diets in animal welfare density (negative control [NC]), basal diet in high stocking density (positive control [PC]), PC + 0.04% essential oil (ES1), PC + 0.08% essential oil (ES2), PC + 0.10% bitter citrus extract & essential oil (CES1), PC + 0.20% bitter citrus extract & essential oil (CES2), PC + 0.05% grape pomace extract (GP1), PC + 0.10% grape pomace extract (GP2). The reduction of space allowance decreased (p < 0.05) average daily gain, feed efficiency, and digestibility of dry matter, crude protein, and gross energy. Also, the fecal score of PC groups increased (p < 0.05) compared with other groups. Basic behaviors (feed intake, standing, lying) were inactive (p < 0.05) and singularity behavior (biting) was increased (p < 0.10) under high stocking density. There was no difference in blood profile. However, the supplementation of PFA alleviated the negative effects such as reduced growth performance, nutrient digestibility, and some increasing stress indicators in th blood (cortisol) and animal behavior (biting). In conclusion, the negative effect of high stocking density was most effectively mitigated by the normal dosage of the mixture of bitter citrus extract and essential oil additive (CES1).
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This study was conducted to investigate the effects of supplementing different ratios of phytogenic feed additives (PFA) to weaned pigs challenged with pathogenic Escherichia coli on growth performance, nutrient digestibility, intestinal barrier integrity, and immune response, and to determine the optimal mixing ratio for post-weaning diarrhea (PWD) prevention. A total of 48 4-wk-old weaned pigs with initial body weight of 8.01 ± 0.39 kg were placed in individual metabolic cages, and then randomly assigned to eight treatment groups. The eight treatments were as follows: a basal diet without E. coli challenge (negative control, NC), a basal diet with E. coli challenge (positive control, PC), PC with supplementing 0.1% mixture of 20% bitter citrus extract (BCE), 10% microencapsulated blend of thymol and carvacrol (MEO), and 70% excipient (T1), PC with supplementing 0.1% mixture of 10% MEO, 20% premixture of grape seed and grape marc extract, green tea, and hops (PGE), and 60% excipient (T2), PC with supplementing 0.1% mixture of 10% BCE, 10% MEO, 10% PGE, and 70% excipient (T3), PC with supplementing 0.1% mixture of 20% BCE, 20% MEO, and 60% excipient (T4), PC with supplementing 0.1% mixture of 20% MEO, 20% PGE, and 60% excipient (T5), and PC with supplementing 0.1% mixture of 10% BCE, 20% MEO, 10% PGE, and 60% excipient (T6). The experiments progressed in 16 days, including 5 days before and 11 days after the first E. coli challenge (day 0). In the E. coli challenge treatments, all pigs were orally inoculated by dividing a total of 10 mL of E. coli F 18 for three consecutive days from day 0 postinoculation (PI). Compared with the PC group, the PFA2 and PFA6 groups significantly increased (P < 0.05) feed efficiency and decreased (P < 0.05) diarrhea during the entire period. At day 11 PI, the PFA6 group significantly improved (P < 0.05) gross energy digestibility compared to the PFA1 group. The PFA6 group significantly decreased (P < 0.05) tumor necrosis factor α (TNF-α) and interleukin-6 in serum and increased (P < 0.05) the villus height to crypt depth ratio (VH:CD). The PFA2 significantly decreased (P < 0.05) the relative protein expression of calprotectin in the ileum. In conclusion, improvements in growth performance, diarrhea reduction, and immunity enhancement are demonstrated when 10% BCE, 20% MEO, 10% PGE, and 60% excipient are mixed.
Phytogenic feed additives (PFA) include various herbs and spices, such as essential oils and polyphenols. Flavonoids and polyphenols contained in PFA are generally known to have antioxidant and antibacterial actions and based on this, PFA is considered an alternative to antibiotics in the swine industry. Pathogenic Escherichia coli infection is one of the most important causes of post-weaning diarrhea (PWD) in pigs. PWD causes intestinal damage, which leads to severe diarrhea, reduced growth performance, and mortality in weaned pigs, resulting in significant financial loss to the swine industry. Therefore, this study was conducted to investigate the effects of supplementing different ratios of PFA to weaned pigs challenged with E. coli and determine the optimal mixing ratio for PWD prevention. Our study results showed that growth performance was improved when supplementing a mixture of 10% bitter citrus extract (BCE), 20% microencapsulated blend of thymol and carvacrol (MEO), 10% premixture of grape seed and grape marc extract, green tea, and hops (PGE), and 60% excipient. Also, the effect of improving the immune response and intestinal morphology was shown. In conclusion, a mixture of 10% BCE, 20% MEO, 10% PGE, and 60% excipients is considered the optimal mixing ratio.
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Infecções por Escherichia coli , Doenças dos Suínos , Suínos , Animais , Escherichia coli , Desmame , Infecções por Escherichia coli/prevenção & controle , Infecções por Escherichia coli/veterinária , Excipientes , Diarreia/prevenção & controle , Diarreia/veterinária , Dieta/veterinária , Nutrientes , Imunidade , Ração Animal/análise , Doenças dos Suínos/prevenção & controleRESUMO
BACKGROUND: This study was conducted to investigate the effects of each phytogenic feed additive (PFA; PFA1, bitter citrus extract; PFA2, a microencapsulated blend of thymol and carvacrol; PFA3, a mixture of bitter citrus extract, thymol, and carvacrol; PFA4, a premixture of grape seed, grape marc extract, green tea, and hops; PFA5, fenugreek seed powder) on the growth performance, nutrient digestibility, intestinal morphology, and immune response in weaned pigs infected with Escherichia coli (E. coli). RESULTS: A total of 63 4-week-old weaned pigs were placed in individual metabolic cages and assigned to seven treatment groups. The seven treatments were as follows: 1) NC; basal diet without E. coli challenge, 2) PC; basal diet with E. coli challenge, 3) T1; PC + 0.04% PFA1, 4) T2; PC + 0.01% PFA2, 5) T3; PC + 0.10% PFA3, 6) T4; PC + 0.04% PFA4, 7) T5; PC + 0.10% PFA5. The experiments lasted in 21 d, including 7 d before and 14 d after the first E. coli challenge. In the E. coli challenge treatments, all pigs were orally inoculated by dividing a total of 10 mL of E. coli F18 for 3 consecutive days. The PFA-added groups significantly increased (P < 0.05) average daily gain and feed efficiency and decreased (P < 0.05) the fecal score at d 0 to 14 post-inoculation (PI). Tumor necrosis factor α was significantly lower (P < 0.05) in the PFA-added groups except for T1 in d 14 PI compared to the PC treatment. The T3 had a higher (P < 0.05) immunoglobulin G and immunoglobulin A concentration compared to the PC treatment at d 7 PI. Also, T3 showed significantly higher (P < 0.05) villus height:crypt depth and claudin 1 expression in ileal mucosa, and significantly down-regulated (P < 0.05) the expression of calprotectin compared to the PC treatment. CONCLUSIONS: Supplementation of PFA in weaned pigs challenged with E. coli alleviated the negative effects of E. coli and improved growth performance. Among them, the mixed additive of bitter citrus extract, thymol, and carvacrol showed the most effective results, improving immune response, intestinal morphology, and expression of tight junctions.
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Arrays of high-index dielectric nanoparticles supporting both electrical and magnetic resonances have gained increasing attention for their excellent light-trapping (LT) effects, thus greatly improving the performance of ultrathin solar cells. This work explores front-located, high-index dielectric subwavelength nanosphere arrays as an efficient and broadband LT structure patterned on top of an ultrathin perovskite solar cell (PSC) for a greatly enhanced absorption. Combined strong light scattering and anti-reflection properties achieved by optimized geometrical parameters of the LT structure lead to a broadband absorption enhancement in the ultrathin thickness of a photoactive layer (100 nm) yielding the short-circuit current density (Jsc) of 18.7 mA/cm2, which is 31.7% higher than that of a planar counterpart. Moreover, effects of the LT structure on far-field radiation patterns, scattering cross-sections, multipoles' contributions, and asymmetry parameters along with the incidence angle and polarization dependence are investigated. The present strategy could be applied to diverse applications, such as other ultrathin or semitransparent solar cells, absorbers and photodetectors.
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As the clinical failure of glioblastoma treatment is attributed by multiple components, including myelin-associated infiltration, assessment of the molecular mechanisms underlying such process and identification of the infiltrating cells have been the primary objectives in glioblastoma research. Here, we adopted radiogenomic analysis to screen for functionally relevant genes that orchestrate the process of glioma cell infiltration through myelin and promote glioblastoma aggressiveness. The receptor of the Nogo ligand (NgR1) was selected as the top candidate through Differentially Expressed Genes (DEG) and Gene Ontology (GO) enrichment analysis. Gain and loss of function studies on NgR1 elucidated its underlying molecular importance in suppressing myelin-associated infiltration in vitro and in vivo. The migratory ability of glioblastoma cells on myelin is reversibly modulated by NgR1 during differentiation and dedifferentiation process through deubiquitinating activity of USP1, which inhibits the degradation of ID1 to downregulate NgR1 expression. Furthermore, pimozide, a well-known antipsychotic drug, upregulates NgR1 by post-translational targeting of USP1, which sensitizes glioma stem cells to myelin inhibition and suppresses myelin-associated infiltration in vivo. In primary human glioblastoma, downregulation of NgR1 expression is associated with highly infiltrative characteristics and poor survival. Together, our findings reveal that loss of NgR1 drives myelin-associated infiltration of glioblastoma and suggest that novel therapeutic strategies aimed at reactivating expression of NgR1 will improve the clinical outcome of glioblastoma patients.
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Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Glioblastoma/metabolismo , Glioblastoma/patologia , Bainha de Mielina/metabolismo , Receptor Nogo 1/metabolismo , Animais , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Proteína 1 Inibidora de Diferenciação/metabolismo , Proteínas Inibidoras de Diferenciação/metabolismo , Camundongos Endogâmicos BALB C , Bainha de Mielina/patologia , Proteases Específicas de Ubiquitina/metabolismoRESUMO
[Purpose] This study was conducted to determine whether acute aerobic exercise (climbing) is associated with changes in the dietary intake pattern. [Subjects and Methods] Food intake and physical activity data for 15 female college students were sampled for 3 days and categorized according to routine activity or high-intensity activity such as hiking. Nutrient intake based on the data was analyzed using a nutrition program. [Results] Carbohydrate and protein intake was significantly decreased after exercise compared to before acute aerobic exercise, but lipid intake showed no significant difference. Calorie intake was significantly decreased after exercise compared to before exercise; however, calorie consumption was significantly increased after exercise. [Conclusion] Aerobic exercise causes a decrease in total calories by inducing reduction in carbohydrate and protein intake. Therefore, aerobic exercise is very important for weight (body fat) control since it causes positive changes in the food intake pattern in female students.
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BACKGROUND/AIMS: Surgical site infection (SSI) is a major complication after radical neck dissection (RND) in patients with head and neck cancer (HNC). We investigated the incidence, risk factors, and etiology of SSI among patients who underwent RND. METHODS: A retrospective cohort study was performed on HNC patients, excluding those with thyroid cancer, who underwent first RND at a teaching hospital between January 2006 and June 2010. Medical records were collected and analyzed to evaluate the risk factors and microbiological etiologies. RESULTS: A total of 370 patients underwent first RND. The overall incidence of SSI was 19.7% (73/370). Multivariate analysis showed that male sex (odds ratio [OR], 4.281; p = 0.004), cardiovascular diseases (OR, 1.941; p = 0.020), large amount of blood loss during surgery (OR, 4.213; p = 0.001), and surgery lasting longer than 6 hours (OR, 4.213; p = 0.002) were significantly associated with SSI. The most common causative pathogen was Staphylococcus aureus (32.6%), and 93.2% of S. aureus isolates were methicillin-resistant. Klebsiella pneumoniae (13/92, 14.1%), Pseudomonas aeruginosa (11/92, 12.0%), and Enterococcus species (11/92, 12.0%) were also frequently detected. CONCLUSIONS: Based on our results, we predict that certain groups of patients are at high risk for SSIs after major HNC surgery. Preventive measures or close monitoring in these patients may be required to reduce the likelihood of postoperative SSIs. Furthermore, even though additional research is required, we would consider changing the prophylactic antibiotic regimens according to the causative organisms.
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Neoplasias de Cabeça e Pescoço/cirurgia , Esvaziamento Cervical/efeitos adversos , Infecção da Ferida Cirúrgica/microbiologia , Idoso , Técnicas Bacteriológicas , Perda Sanguínea Cirúrgica , Doenças Cardiovasculares/complicações , Distribuição de Qui-Quadrado , Feminino , Humanos , Incidência , Modelos Logísticos , Masculino , Prontuários Médicos , Pessoa de Meia-Idade , Análise Multivariada , Razão de Chances , Duração da Cirurgia , República da Coreia , Estudos Retrospectivos , Fatores de Risco , Fatores Sexuais , Infecção da Ferida Cirúrgica/diagnóstico , Infecção da Ferida Cirúrgica/epidemiologia , Resultado do TratamentoRESUMO
While cell culture-based technology has been recently used for manufacturing influenza vaccines, currently available seed viruses are mostly egg-derived reassortants that are egg-adapted to achieve high virus growth in eggs. For use as viruses for cell culture-based influenza vaccine manufacturing, egg-adapted viral seeds may undergo several passages in manufacturing cell lines. However, the suitability of such cell-passaged viruses for vaccine production remains largely unelucidated. In this study, influenza viruses produced in suspension Madin-Darby canine kidney (MDCK) cell cultures were compared to those produced in embryonated hen's eggs for manufacturing MDCK cell culture-based influenza vaccines through comparability studies of virus productivity and vaccine immunogenicity. The results indicate no change in the amino acid sequence of the main antigens, including hemagglutinin (HA) and neuraminidase (NA), of cell-passaged viruses after three passages in suspension MDCK cells. In lab-scale (3-L) single-use bioreactors, suspension MDCK culture supernatants inoculated with cell-passaged viruses were found to show higher virus productivity, suspension MDCK culture supernatants inoculated with egg-passaged viruses, in respect to the HA titers and HA contents determined by single radial immunodiffusion. Finally, comparable hemagglutination inhibition and influenza-specific IgG titers were determined in the mice immunized with cell culture-based vaccines produced with cell- or egg-passaged viruses. These results indicate that MDCK cell-passaged viruses from egg-adapted viruses, as well as egg-derived seed virus, are suitable for MDCK cell culture-based influenza vaccine production.
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Antígenos Virais/imunologia , Galinhas/imunologia , Vacinas contra Influenza/imunologia , Influenza Humana/prevenção & controle , Orthomyxoviridae/imunologia , Cultura de Vírus/métodos , Animais , Técnicas de Cultura de Células , Galinhas/virologia , Cães , Ovos , Feminino , Testes de Inibição da Hemaglutinação , Hemaglutininas/imunologia , Humanos , Imunização , Influenza Humana/imunologia , Células Madin Darby de Rim Canino , Camundongos , Neuraminidase/imunologiaRESUMO
Light-emitting 5,6,11,12-tetraphenylnaphthacene (rubrene) nanoparticles (NPs) prepared by a reprecipitation method were treated hydrothermally. The diameters of hydrothermally treated rubrene NPs were changed from 100 nm to 2 µm, depending on hydrothermal temperature. Photoluminescence (PL) characteristics of rubrene NPs varied with hydrothermal temperatures. Luminescence of pristine rubrene NPs was yellow-orange, and it changed to blue as the hydrothermal temperature increased to 180°C. The light-emitting color distribution of the NPs was confirmed using confocal laser spectrum microscope. As the hydrothermal temperature increased from 110°C to 160°C, the blue light emission at 464 to approximately 516 nm from filtered-down NPs was enhanced by H-type aggregation. Filtered-up rubrene NPs treated at 170°C and 180°C exhibited blue luminescence due to the decrease of intermolecular excimer densities with the rapid increase in size. Variations in PL of hydrothermally treated rubrene NPs resulted from different size distributions of the NPs.
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Functionalization of light-emitting poly(3-methylthiophene) (P3MT) nanowires (NWs) with probe-DNA (p-DNA) and their label-free recognition of target-DNA (t-DNA) were correlated quantitatively with both the photoluminescence (PL) color and intensity of P3MT NWs.
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Técnicas de Química Analítica/instrumentação , DNA/análise , Luz , Nanofios/química , Polímeros/química , Tiofenos/química , Óxido de Alumínio/química , Benzenossulfonatos/química , DNA/química , Eletroquímica , Eletrodos , Medições Luminescentes , Compostos de Amônio Quaternário/químicaRESUMO
PURPOSE: The purpose of this study was to develop a population pharmacokinetic (PPK) model of glimepiride and to investigate the influence of genetic polymorphisms in CYP2C9 on the PPK of glimepiride in healthy Korean subjects. METHODS: Serum data after a single oral dose of 2 mg of glimepiride in 177 healthy male Korean subjects (CYP2C9*1*1: 163 subjects, *1/*3: 14 subjects) were used. We estimated the PPK of glimepiride using a nonlinear mixed effects modeling (NONMEM) method and explored the possible influence of genetic polymorphisms in CYP2C9 on the PPK of glimepiride. RESULTS: The disposition of glimepiride was best described with a two-compartment model with a Weibull-type absorption and first-order elimination. The visual predictive check indicated that the pharmacokinetic profile of glimepiride was adequately described by the proposed PPK model. The CYP2C9 genotypes as covariate significantly (P < 0.001) influenced the apparent oral clearance (CL/F) of glimepiride. The estimated CL/F of glimepiride was higher (1.60-fold) in CYP2C9*1/*1 subjects than in CYP2C9*1/*3 subjects. CONCLUSIONS: This study indicates that genetic polymorphisms of CYP2C9 influence the substantial interindividual variability in the disposition of glimepiride, and these polymorphisms may affect the clinical response to glimepiride therapy.
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Hidrocarboneto de Aril Hidroxilases/genética , Hipoglicemiantes/farmacocinética , Modelos Biológicos , Polimorfismo de Fragmento de Restrição , Compostos de Sulfonilureia/farmacocinética , Absorção , Administração Oral , Adulto , Estudos Cross-Over , Citocromo P-450 CYP2C9 , Humanos , Hipoglicemiantes/sangue , Masculino , Taxa de Depuração Metabólica , Mucosa Bucal/metabolismo , Reação em Cadeia da Polimerase , República da Coreia , Compostos de Sulfonilureia/sangue , Equivalência Terapêutica , Adulto JovemRESUMO
Poly(3-hexylthiophene) (P3HT) nanoparticles (NPs) were prepared by a reprecipitation method. Hydrothermal processing applied external pressure to the pristine P3HT NPs at temperatures ranging from 60 to 150 °C. Optical absorption and photoluminescence (PL) spectra for the hydrothermally treated P3HT NPs varied markedly with the processing temperature. With increasing treatment temperature, the absorption peak broadened and the peak position shifted from 510 to 623 nm; moreover, the intensity ratio of the 0-1 to 0-0 emission varied. These changes were caused by interactions between the P3HT main chains and alkyl side groups and conformational modifications induced by the high pressure during the hydrothermal process. The evolution of the optical absorption spectra of the P3HT NPs during the hydrothermal processing was strongly correlated with the variation of PL excitation spectra and with the PL emission spectra of a single NP.
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We report on the light-emitting color barcode nanowires (LECB-NWs), which were fabricated by alternating the electrochemical polymerization of light-emitting polymers with various luminescence colors and efficiencies. The nanoscale photoluminescence characteristics of LECB-NWs were investigated using a laser confocal microscope with a high spatial resolution. The alternating light emissions of the LECB-NWs showed orange-yellow, red, and green colors due to the serial combination of poly(3-butylthiophene), poly(3-methylthiophene), and poly(3,4-ethylenedioxythiophene), respectively, with distinct luminescence intensities. The optical detection sensitivity and stability of LECB-NWs have been enhanced through a nanoscale Cu metal coating onto the NWs, based on surface plasmon resonance coupling and protection against oxidation. The flexibility of the LECB-NWs has been investigated through the folding and unfolding of the NWs by an applied nanotip impetus. The flexible LECB-NWs can be used as highly sensitive optical identification nanosystems for nanoscale or microscale products with complex physical shapes.
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Processamento Eletrônico de Dados/métodos , Luz , Nanotecnologia/métodos , Nanofios/química , Fenômenos Ópticos , Polímeros/química , Cor , Cobre/química , Elasticidade , Transferência de EnergiaRESUMO
Human astrovirus (HAstV) is the second most important cause of viral diarrhea and acute gastroenteritis in infants under five. However, determination of the infectivity of clinical isolates is difficult, and the replication cycle of HAstV is not yet fully understood. In this study, it was attempted to detect negative-sense (-)RNAs generated during the replication of RNA viruses. We used clinical isolates of HAstV to infect CaCo-2 cells. Reverse transcription using only a sense primer followed by PCR using both sense and antisense primers showed that (-)RNAs were first detected in CaCo-2 cells between 9 and 12 h postinfection (p.i.). However, these (-)RNAs were not detected when cells were treated with the protein synthesis inhibitor cycloheximide during HAstV infection. Next, RT with only an antisense primer followed by PCR was performed to detect (+)RNA of HAstVs after production of (-)RNAs during replication. RT-PCR results using the antisense primer revealed that the amount of (+)RNA began to increase starting 9 h p.i., indicating an accumulation of the newly synthesized (+)RNA genome. Cycloheximide was observed to abrogate the increase of newly made (+)RNA during HAstV infection. In conclusion, the use of sense or antisense primers during the RT reaction together with cycloheximide enabled us to quantitatively detect (-)RNAs, and this proved to be an useful tool in understanding the replication cycle of HAstV.
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Infecções por Astroviridae/virologia , Mamastrovirus/isolamento & purificação , Mamastrovirus/fisiologia , RNA Viral/genética , Replicação Viral , Infecções por Astroviridae/diagnóstico , Células CACO-2 , Primers do DNA/genética , Fezes/virologia , Humanos , Mamastrovirus/genética , Reação em Cadeia da PolimeraseRESUMO
Pi-conjugated organic systems have been used as optoelectronic and sensing materials due to their characteristics of efficient light emission or absorption, and p-type charge transport. The hybrid nanostructures of pi-conjugated organic systems with nanoscale metals offer surface plasmon (SP)-enhanced luminescence, which can be applied to organic-based optoelectronics, photonics, and sensing. Various hybrid nanostructures using light-emitting polymers with nanoscale metals have been fabricated and have shown considerable enhancement of photoluminescence efficiency due to energy and charge transfer effects in SP resonance coupling. In this tutorial review, recent conceptual and technological achievements in light-emitting polymers-based hybrid nanostructures are described.
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Luminescência , Nanoestruturas/química , Nanotecnologia/métodos , Ressonância de Plasmônio de Superfície/métodos , Quimera , Luz , Metais/química , Óptica e Fotônica/métodos , Compostos Orgânicos , Polímeros/químicaRESUMO
Traditionally primers for PCR detection of viruses have been selected from genomic sequence of single or representative viral strain. However, high mutation rate of viral genomes often results in failure in detecting viruses in clinical and environmental samples. Thus, it seems necessary to consider primers designed from multiple viral sequences in order to improve detection of viral variants. Matchup is a program intended to select universal primers from multiple sequences. We designed using Matchup program primer pairs for HBV detection from 691 full genomic HBV DNA sequences available from NCBI GenBank database. Thousands of primer candidates were initially extracted and these were sequentially filtered down to 5 primer pairs. These primer pairs were tested by PCR using 5 HBV Korean HBsAg(+) patient sera, and eventually one universal primer pair was selected and named MUW (multiple-universal-worldwide). This primer pair, 3 HBV reference primer pairs reported by others and 1 commercial primer pair were compared using 86 HBV HBsAg(+) sera from Korean and Vietnamese patients. The detection rate for MUW primer pair was 72.1%, much greater than those obtained by reference and commercial primers (32.5 to 40.7%). The superiority of MUW primer pair appeared to be correlated with the conserved sequences of the forward primer binding sites and primer quality score. These results suggest that the universal primers designed by the Matchup program from multiple sequences could be useful in detecting viruses from clinical samples.
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Primers do DNA/genética , DNA Viral/genética , Vírus da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA/métodos , Sequência de Bases , DNA Viral/sangue , Bases de Dados Genéticas , Variação Genética , Humanos , Dados de Sequência Molecular , Mutação , Carga ViralRESUMO
Cyclooxygenase (COX)-2 plays an important role in inflammatory reaction and human cytomegalovirus (HCMV) is known to induce COX-2 gene expression. However, little is known for the mechanism of COX-2 gene expression by HCMV. In this study, three recently isolated HCMV strains including TB40/E and clinical isolates from Korean patients as well as highly laboratory adapted strain AD169 were used to stimulate COX-2 induction. Western blot analyses revealed that recently isolated HCMV strains induced COX-2 expression in human foreskin fibroblast (HFF) cells to higher levels than lab-adapted strain AD169. HCMV gene expression was not required for COX-2 induction since UV-inactivated virus was able to stimulate COX-2 gene expression during immediate early infection. Moreover, when HCMV-infected HFF cells were transfected with small interfering RNA for major immediate early (MIE) genes, HCMV-induced COX-2 expression was not abrogated, suggesting that HCMV MIE genes are not needed for COX-2 expression. In order to understand the signal pathways involved in COX-2 gene expression induced by HCMV, selective inhibitors were used. When HCMV-infected HFF cells were treated with epidermal growth factor receptor (EGFR) kinase inhibitors and c-Raf/mitogen-activated protein kinases (MAPK, MEK) 1/2 inhibitors, HCMV-induced COX-2 expression was diminished. However, COX-2 expression by HCMV infection was not inhibited if HCMV-infected HFF cells were treated with inhibitors of phosphatidylinositol-3-kinase, c-Jun N-terminal kinase, nuclear factor kappa B, p-38 MAPK and Rho-associated protein kinase. We concluded that EGFRK, Raf, MEK1/2 and extracellular signal-regulated kinases (ERK 1/2) pathway may participate in the COX-2 mediated inflammatory response to HCMV infection.
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Ciclo-Oxigenase 2/biossíntese , Citomegalovirus/patogenicidade , Interações Hospedeiro-Patógeno , Western Blotting , Células Cultivadas , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/virologia , Receptores ErbB/metabolismo , Humanos , Coreia (Geográfico) , MAP Quinase Quinase 1/metabolismo , MAP Quinase Quinase 2/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Proteínas Proto-Oncogênicas c-raf/metabolismoRESUMO
Complex nanoparticles (NPs) of poly(2-methoxy-5-(2'-ethylhexyloxy)-p-phenylene vinylene) (MEH-PPV) NP adsorbed with Au NPs (MEH-PPV/Au NPs) were fabricated through a reprecipitation method. The formation of MEH-PPV/Au NP complexes was confirmed through high-resolution transmission electron microscopy and Fourier transform infrared experiments. The laser confocal microscope photoluminescence (PL) efficiency of the complex MEH-PPV/Au single NP dramatically increased compared with that of the MEH-PPV single NP without Au NPs, which was directly confirmed through color charge-coupled device images. The enhanced PL efficiency of the MEH-PPV/Au NP complex might have originated from the energy transfer effect in a surface plasmon resonance coupling between a MEH-PPV NP and Au NPs. The strong local field enhancement due to nanogaps between Au NPs in the background of a light-emitting MEH-PPV NP might be another origin of the PL enhancement of the NP complex, as supported by finite difference time domain calculations. We also observed the blue shift of the PL peaks of the single MEH-PPV and MEH-PPV/Au NP, compared with the solution PL peaks of those NPs.
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Molecular phylogenetic studies of the HIV-1 isolated from Koreans have suggested the presence of the so-called "Korean clade", which can be defined as a cluster free of foreign isolates. The Korean clade accounts for more than 60% of Korean isolates and exerts characteristic amino acid sequences. Thus, it is merited to estimate when this Korean clade first emerged in order to understand the evolutionary pattern of the Korean clade. We analyzed and reconstructed the most recent common ancestor (MRCA) sequences from nef (n=229) and vif (n=179) Korean clade sequences. Linear regression analyses of sequence divergence estimates were plotted against sampling years to infer the year in which there was zero divergence from the MRCA sequences. MRCA sequences suggested the Korean clade was first emerged around 1984, before the first detection of HIV-1 in Korea in 1985. Further studies on synonymous and nonsynonymous substitution rates suggested positive selection event for the Korean clade, while other subtype B had undergone negative to neutral evolution.