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1.
Int J Antimicrob Agents ; 63(1): 107011, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37863340

RESUMO

OBJECTIVES: Alternation of the colistin resistance-regulating two-component regulatory system (crrAB) is a colistin-resistance mechanism in Klebsiella pneumoniae (K. pneumoniae), but its role in bacteria is not fully understood. METHODS: Twelve colistin-susceptible K. pneumoniae clinical isolates were included in this study: six crrAB-positive and six crrAB-negative. We deleted the crrAB genes from two crrAB-positive isolates and complemented them. We measured the growth yields by determining growth curves in lysogeny broth and minimal media with or without Fe2+. In vitro selection rates for colistin resistance were determined by exposure to colistin, and survival rates against high concentrations of colistin (20 mg/L) at the early stage of growth (20 min) were investigated. Virulence was determined using a serum bactericidal assay and Galleria mellonella larval infection. RESULTS: The presence of crrAB was not associated with colistin resistance and did not increase the in vitro selection rate of colistin resistance after exposure. The growth yield of crrAB-positive isolates was higher in lysogeny broth media and increased when Fe2+ was added to minimal media. The crrAB-positive isolates showed higher survival rates in the early stages of exposure to high colistin concentrations. Decreased serum resistance was identified in the crrAB-deleted mutants. More G. mellonella larvae survived when infected by crrAB-deleted mutants, and higher survival rates of bacteria were identified within the larvae infected with wild-type than crrAB-deletant isolates. CONCLUSION: Through rapid response to external signals, crrAB would provide advantages for K. pneumoniae survival by increasing the final growth yield and initial survival against colistin treatment. This may partly contribute to the bacterial virulence.


Assuntos
Colistina , Infecções por Klebsiella , Animais , Colistina/farmacologia , Colistina/uso terapêutico , Klebsiella pneumoniae , Virulência , Proteínas de Bactérias/genética , Farmacorresistência Bacteriana/genética , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Larva , Testes de Sensibilidade Microbiana , Infecções por Klebsiella/tratamento farmacológico , Infecções por Klebsiella/microbiologia
2.
J Glob Antimicrob Resist ; 36: 45-49, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38128724

RESUMO

OBJECTIVES: Genetic changes in Mycobacterium abscessus during antibiotic treatment are not fully understood. This study aimed to investigate the genetic changes in M. abscessus in patients receiving antibiotic treatment, and their clinical implications. METHODS: Pretreatment and 12-month post-treatment M. abscessus isolates were obtained from patients with M. abscessus pulmonary disease. Isolates from each time point were separated into six groups based on their distinctive morphological characteristics. Twenty-four isolates, comprising 12 from patient A exhibiting progressive disease and 12 from patient B demonstrating stable disease, underwent sequencing. Subsequently, minimal inhibitory concentrations (MICs) for the administered antibiotics were measured. RESULTS: Persistent infection with a single strain was observed in patients A and B. During 12 months of treatment, MICs for administered drugs did not generally change over time in either patient and single nucleotide variations (SNV) associated with antimicrobial resistance (rrl, rrs, erm(41), gyrA, gyrB, whiB7 and hflX) were not mutated. Although not significant, 47 and 52 non-synonymous SNVs occurred in M. abscessus from patients A and B, respectively, and the accumulation of these SNVs differed in patients A and B, except for five SNVs. The most variable positions were within a probable NADH-dependent glutamate synthase gene and a putative YrbE family protein gene in patients A and B, respectively. CONCLUSIONS: Persistent infections by a single strain of M. abscessus were observed in two patients with different clinical courses. Genetic changes in M. abscessus during antibiotic treatment were relatively stable in these patients. CLINICAL TRIALS IDENTIFIER: NCT01616745 (ClinicalTrials.gov ID).


Assuntos
Pneumopatias , Mycobacterium abscessus , Humanos , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Testes de Sensibilidade Microbiana , Mycobacterium abscessus/genética
3.
Crit Care Med ; 48(8): e706-e714, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32697512

RESUMO

OBJECTIVE: To determine the effects of adjunctive therapeutic hypothermia, by comparing hyperbaric oxygen therapy versus hyperbaric oxygen therapy combined with therapeutic hypothermia in acute severe carbon monoxide poisoning. DESIGN: Retrospective analysis of data from our prospectively collected carbon monoxide poisoning registry. SETTING: A single academic medical center in Wonju, Republic of Korea. PATIENTS: Patients with acute severe carbon monoxide poisoning older than 18 years. Acute severe carbon monoxide poisoning was defined as mental status showing response to painful stimulus or unresponsive at the emergency department, and a continuation of this depressed mental status even after the first hyperbaric oxygen therapy. Patients were classified into the no-therapeutic hypothermia and therapeutic hypothermia groups. Hyperbaric oxygen therapy was performed up to twice within 24 hours after emergency department arrival, whereas therapeutic hypothermia was performed at a body temperature goal of 33°C for 24 hours using an endovascular cooling device after the first hyperbaric oxygen therapy. INTERVENTIONS: Hyperbaric oxygen therapy versus hyperbaric oxygen therapy combined with therapeutic hypothermia. MEASUREMENTS AND MAIN RESULTS: We investigated the difference in the Global Deterioration Scale score at 1 and 6 months after carbon monoxide exposure, between the no-therapeutic hypothermia and therapeutic hypothermia groups. Global Deterioration Scale scores were classified as follows: 1-3 points (favorable neurocognitive outcome) and 4-7 points (poor neurocognitive outcome). During the study period, 37 patients were treated for acute severe carbon monoxide poisoning, with 16 and 21 patients in the no-therapeutic hypothermia and therapeutic hypothermia groups, respectively. The therapeutic hypothermia group demonstrated significantly higher number of patients with favorable outcomes (p = 0.008) at 6 months after carbon monoxide exposure and better improvement of the 6-month Global Deterioration Scale score than the 1-month score (p = 0.006). CONCLUSIONS: Our data suggest that in acute severe carbon monoxide poisoning, patients who were treated using therapeutic hypothermia combined with hyperbaric oxygen therapy had significantly more favorable neurocognitive outcomes at 6 months after carbon monoxide exposure than those treated with hyperbaric oxygen therapy alone.


Assuntos
Intoxicação por Monóxido de Carbono/terapia , Oxigenoterapia Hiperbárica/métodos , Hipotermia Induzida/métodos , Adolescente , Adulto , Idoso , Terapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto Jovem
4.
Undersea Hyperb Med ; 46(2): 135-143, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31051058

RESUMO

Purpose: Hyperbaric medicine is nascent in Korea when compared to other developed countries, such as the United States and Japan. Our facility has been managed by physicians with certifications from the Undersea and Hyperbaric Medical Society (UHMS) and National Oceanic and Atmospheric Administration in diving and clinical diseases since October 2016. This study was conducted to share similar issues that are encountered during the establishment of a program in a new area through our experiences in the operation of a hyperbaric oxygen (HBO2) therapy center. Methods: In this retrospective observational study we collected data on HBO2 patients treated at our center between October 2016 and June 2018 after HBO2 was conducted by HBO2-certified physicians. We then compared demographic data of patients with data from January 2011 to September 2015 - before HBO2 operations were conducted by HBO2-certified physicians. Result: A total of 692 patients received 5,130 treatments. Twelve indicated diseases were treated using HBO2 therapy. Fifty-six critically ill patients with intubation received HBO2. Although two patients experienced seizure due to oxygen toxicity during the study period, certified physicians and inside attendant took immediate corrective action. Conclusion: After the establishment of the HBO2 center operated by physicians with certification, more patients, including critically ill patients, received HBO2 safely for various diseases. In order to improve the practice of hyperbaric medicine in Korea, the Korean Academy of Undersea and Hyperbaric Medicine (KAUHM), an advanced and well-organized academic society, should communicate often with HBO2 centers, with the aim to set Korean education programs at UHMS course levels and increase reimbursement for HBO2 therapy.


Assuntos
Hospitais Especializados/estatística & dados numéricos , Oxigenoterapia Hiperbárica/estatística & dados numéricos , Desenvolvimento de Programas , Idoso , Intoxicação por Monóxido de Carbono/terapia , Certificação , Estado Terminal/terapia , Doença da Descompressão/terapia , Feminino , Perda Auditiva Neurossensorial/terapia , Perda Auditiva Súbita/terapia , Hospitais Especializados/organização & administração , Humanos , Oxigenoterapia Hiperbárica/efeitos adversos , Oxigenoterapia Hiperbárica/instrumentação , Masculino , Pessoa de Meia-Idade , Objetivos Organizacionais , Oxigênio/efeitos adversos , República da Coreia , Estudos Retrospectivos , Suspensão de Tratamento/estatística & dados numéricos
5.
J Sci Food Agric ; 99(2): 711-718, 2019 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-29971800

RESUMO

BACKGROUND: In recent years, protected crop production using plant factories to produce high-value crops with greater functional components has become more popular in many countries. The quantification of the components, however, is mainly conducted by laboratory analyses, which are both time- and labor-consuming. The present study aimed to investigate the potential of a non-destructive diffuse reflectance spectroscopy technique for estimating functional components (i.e. glucosinolates, amino acids, sugars and carotenoids) in the leaves of Chinese cabbage grown in a plant factory. RESULTS: From the overall analysis, better estimations were obtained using the partial least square regression procedure. The important wavelengths for each functional component were identified mainly in the ultraviolet-visible regions. Identified wavelengths were 317, 390, 888 and 940 nm for sugars; 520 and 960 nm for amino acids; 385, 860 and 945 nm for glucosinolates; and 454, 472 and 530 nm for carotenoids. CONCLUSION: Optical reflectance spectroscopy shows potential as a tool for the estimation of functional components in the leaves of Chinese cabbage. The results of the present study provide useful information for the design and application of sensors with respect to on-site quantification of the functional components. © 2018 Society of Chemical Industry.


Assuntos
Brassica/química , Extratos Vegetais/química , Folhas de Planta/química , Análise Espectral/métodos , Carotenoides/química , Glucosinolatos/química
6.
Molecules ; 21(2): 157, 2016 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-26828471

RESUMO

A total of 13 anthocyanins and 33 metabolites; including organic acids, phenolic acids, amino acids, organic compounds, sugar acids, sugar alcohols, and sugars, were profiled in three radish cultivars by using high-performance liquid chromatography (HPLC) and gas chromatography time-of-flight mass spectrometry (GC-TOFMS)-based metabolite profiling. Total phenolics and flavonoids and their in vitro antioxidant activities were assessed. Pelargonidins were found to be the major anthocyanin in the cultivars studied. The cultivar Man Tang Hong showed the highest level of anthocyanins (1.89 ± 0.07 mg/g), phenolics (0.0664 ± 0.0033 mg/g) and flavonoids (0.0096 ± 0.0004 mg/g). Here; the variation of secondary metabolites in the radishes is described, as well as their association with primary metabolites. The low-molecular-weight hydrophilic metabolite profiles were subjected to principal component analysis (PCA), hierarchical clustering analysis (HCA), Pearson's correlation analysis. PCA fully distinguished the three radish cultivars tested. The polar metabolites were strongly correlated between metabolites that participate in the TCA cycle. The chemometrics results revealed that TCA cycle intermediates and free phenolic acids as well as anthocyanins were higher in the cultivar Man Tang Hong than in the others. Furthermore; superoxide radical scavenging activities and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging were investigated to elucidate the antioxidant activity of secondary metabolites in the cultivars. Man Tang Hong showed the highest superoxide radical scavenging activity (68.87%) at 1000 µg/mL, and DPPH activity (20.78%), followed by Seo Ho and then Hong Feng No. 1. The results demonstrate that GC-TOFMS-based metabolite profiling, integrated with chemometrics, is an applicable method for distinguishing phenotypic variation and determining biochemical reactions connecting primary and secondary metabolism. Therefore; this study might provide information on the relationship between primary and secondary metabolites and a synergistic antioxidant ability derived from the secondary metabolites in the radish cultivars.


Assuntos
Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Metabolômica/métodos , Raphanus/química , Antocianinas/isolamento & purificação , Antocianinas/farmacologia , Antioxidantes/química , Cromatografia Líquida de Alta Pressão/métodos , Flavonoides/química , Flavonoides/isolamento & purificação , Flavonoides/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Fenóis/química , Fenóis/isolamento & purificação , Fenóis/farmacologia , Extratos Vegetais/análise , Análise de Componente Principal , Raphanus/classificação
7.
Nat Prod Commun ; 11(9): 1283-1286, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30807023

RESUMO

This study investigated the influence of auxins on the growth of hairy roots and accumulation of anthocyanins, including cyanidin 3-O-glucoside (C3gl) and cyanidin 3-0-rutinoside (C3r), in the hairy root culture of tartary buckwheat cultivar Hokkai TIO. C3gl and C3r contents were evaluated using high- performance liquid chromatograph (HPLC). Four auxins, 2,4-dichlorophenoxyacetic acid (2,4-D), indoleacetic acid (IAA), indolebutyric acid (IBA) and naphthaleneacetic acid (NAA), were added to the medium of the hairy root cultures at diverse concentrations. IAA, IBA and 2,4-D promoted the growth of hairy roots since the dry weight of the roots was slightly higher than or comparable with that of the control. However, NAA at all concentrations suppressed the growth of hairy roots. Generally, auxin treatments resulted in higher accumulation of C3gl and C3r than that of the control except for 2.85 µM IAA and 2.69 µM NAA. The amount of C3gl and C3r after treatment with 4.92 µM IBA was the highest among all treatments and was 3.24 times more than that of the control. Our results suggested that auxins at appropriate concentrations might facilitate hairy root growth of tartary buckwheat and enhance the production of C3gl and C3r.


Assuntos
Antocianinas/biossíntese , Fagopyrum/química , Ácidos Indolacéticos/farmacologia , Raízes de Plantas/química , Fagopyrum/efeitos dos fármacos , Fagopyrum/crescimento & desenvolvimento , Glucosídeos , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos
8.
Nat Prod Commun ; 10(7): 1211-4, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26411013

RESUMO

Radish sprouts and young seedlings are considered important dietary vegetables in Asian countries. In this study, we investigated the levels of glucosinolate and anthocyanin accumulation in radish seedlings in response to light and methyl jasmonate (MeJA) treatments. MeJA facilitated the accumulation of glucosinolate and anthocyanins under light conditions. The glucosinolate and anthocyanin contents in the radish seedlings that were exposed to light after MeJA treatment were higher than those of the seedlings that were grown in the dark without MeJA. At a concentration of 100 µM, MeJA led to the greatest accumulation of the most glucosinolates under both light and dark conditions. Under light conditions, the levels of glucoraphenin, glucoerucin, and glucotropaeolin accumulation were 1.53-, 1.60-, and 1.30-fold higher, respectively, than those of the control. Remarkable accumulations of glucobrassicin were observed under light conditions (4.4-, 6.7-, and 7.8-fold higher than that of the control following the application of 100, 300, and 500 µM MeJA, respectively). The level of cyanidin in the 300 µM MeJA-treated seedlings was double of that in the control without MeJA treatment. The highest level of pelargonidin was observed after treatment with 500 µM MeJA under light conditions; this level was 1.73 times higher than that in the control. A similar trend of anthocyaninaccumulation was observed in the radish seedlings following MeJA treatment under dark conditions, but the levels of anthocyanins were considerably lower in the seedlings that were grown in the dark. Our findings suggest that light and low concentrations of MeJA enhance the accumulations of glucosinolates and anthocyanins during the development of radish seedlings.


Assuntos
Acetatos/metabolismo , Antocianinas/biossíntese , Ciclopentanos/metabolismo , Glucosinolatos/biossíntese , Oxilipinas/metabolismo , Raphanus/metabolismo , Raphanus/efeitos da radiação , Luz , Plântula/metabolismo , Plântula/efeitos da radiação
9.
Phytomedicine ; 22(6): 615-20, 2015 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-26055126

RESUMO

BACKGROUND: The bark of Machilus thunbergii (Lauraceae) has been used as a folk medicine to treat abdominal pain and distension, and leg edema in Korea. Machilin A (MA), a lignan isolated from Machilus thunbergii, exhibits several biological activities including anti-oxidant and stimulatory effects on cell differentiation and proliferation. PURPOSE: Potential drug-interactions with MA via inhibition of cytochrome P450 (CYP) activity in human liver microsomes (HLMs), have not been investigated. STUDY DESIGN: The inhibitory effects of MA on the activities of CYPs were investigated using cocktail probe substrates in pooled HLMs and on human recombinant cDNA-expressed CYP isoforms. METHODS: The nine CYP-specific substrates were incubated in HLM or recombinant cDNA-expressed CYP 1A1, 1A2 and 2B6 with MA. After incubation, the samples were injected onto a C18 column for liquid chromatography-tandem mass spectrometry analysis. To investigate the binding poses between MA and CYP, we carried out structure-based docking simulations by using software and scripts written in-house (ALIS-DOCK; Automatic pLatform for Iterative Structure-based DOCKing). RESULTS: MA strongly inhibited CYP1A2-mediated phenacetin O-deethylation and CYP2B6-mediated bupropion hydroxylation with IC50 values of 3.0 and 3.9 µM, respectively, while it did not significantly inhibit other CYPs. A Dixon plot indicated that MA competitively inhibits CYP1A2 and CYP2B6 with Ki values of 0.71 and 4.1 µM, respectively. CONCLUSION: Overall, this was the first investigation of the inhibitory effects of MA on CYP1A2 and CYP2B6 in HLMs, and it has identified that MA acts via competitive inhibition.


Assuntos
Benzodioxóis/farmacologia , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1A2/metabolismo , Citocromo P-450 CYP2B6/metabolismo , Inibidores das Enzimas do Citocromo P-450/farmacologia , Lauraceae/química , Lignanas/farmacologia , Humanos , Microssomos Hepáticos/enzimologia , Simulação de Acoplamento Molecular , Estrutura Molecular , Casca de Planta/química , Proteínas Recombinantes/metabolismo
10.
Food Chem ; 177: 204-13, 2015 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-25660878

RESUMO

The present study aimed to investigate the effects of different l-phenylalanine (l-Phe) concentrations and various light-emitting diodes (LEDs) on the accumulation of phenolic compounds (chlorogenic acid, vitexin, rutin, quercetin, cyanidin 3-O-glucoside, and cyanidin 3-O-rutinoside) in Tartary buckwheat sprouts. We found that 5mM was the optimum l-Phe concentration for the synthesis of total and individual phenolic compounds. The highest rutin (53.09 mg/g DW) and chlorogenic acid (5.62 mg/g DW) content was observed with Red+Blue and white lights. Comprehensive differences in total and individual anthocyanin content were observed between different lights; however, the total anthocyanin content (9.12 mg/g DW) was 1.5-fold higher in blue light. The expression levels of regulatory genes, such as FtDFR and FtANS, were 7.1-fold higher with l-Phe treatment. Gene expression results showed that the phenolic compounds in Tartary buckwheat sprouts increased with the use of l-Phe and LED lights.


Assuntos
Antocianinas/biossíntese , Fagopyrum/efeitos dos fármacos , Fagopyrum/efeitos da radiação , Flavonoides/biossíntese , Fenilalanina/farmacologia , Fagopyrum/genética , Fagopyrum/metabolismo , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/efeitos da radiação , Luz
11.
Molecules ; 19(11): 17141-53, 2014 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-25347458

RESUMO

Riboflavin (vitamin B2) is the precursor of flavin mononucleotide and flavin adenine dinucleotide-essential cofactors for a wide variety of enzymes involving in numerous metabolic processes. In this study, a partial-length cDNA encoding bifunctional GTP cyclohydrolase II/3,4-dihydroxy-2-butanone-4-phosphate synthase (LcRIBA), 2 full-length cDNAs encoding lumazine synthase (LcLS1 and LcLS2), and a full-length cDNA encoding riboflavin synthase (LcRS) were isolated from Lycium chinense, an important traditional medicinal plant. Sequence analyses showed that these genes exhibited high identities with their orthologous genes as well as having the same common features related to plant riboflavin biosynthetic genes. LcRIBA, like other plant RIBAs, contained a DHBPS region in its N terminus and a GCHII region in its C-terminal part. LcLSs and LcRS carried an N-terminal extension found in plant riboflavin biosynthetic genes unlike the orthologous microbial genes. Quantitative real-time polymerase chain reaction analysis showed that 4 riboflavin biosynthetic genes were constitutively expressed in all organs examined of L. chinense plants with the highest expression levels found in the leaves or red fruits. LcRIBA, which catalyzes 2 initial reactions in riboflavin biosynthetic pathway, was the highest transcript in the leaves, and hence, the richest content of riboflavin was detected in this organ. Our study might provide the basis for investigating the contribution of riboflavin in diverse biological activities of L. chinense and may facilitate the metabolic engineering of vitamin B2 in crop plants.


Assuntos
DNA Complementar/genética , GTP Cicloidrolase/genética , Lycium/genética , Complexos Multienzimáticos/genética , Riboflavina Sintase/genética , Riboflavina/genética , Riboflavina/metabolismo , Sequência de Aminoácidos , Biodiversidade , GTP Cicloidrolase/metabolismo , Genes de Plantas/genética , Lycium/metabolismo , Complexos Multienzimáticos/metabolismo , Peptídeo Sintases/genética , Peptídeo Sintases/metabolismo , Plantas Medicinais/genética , Plantas Medicinais/metabolismo , Riboflavina Sintase/metabolismo , Alinhamento de Sequência , Fosfatos Açúcares/metabolismo
12.
Appl Biochem Biotechnol ; 174(7): 2537-47, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25194705

RESUMO

Differential expression patterns of flavonoid biosynthetic pathway genes in the hairy roots of tartary buckwheat cultivars "Hokkai T8" and "Hokkai T10" were studied over a time course of the light-dark cycle. The Agrobacterium rhizogenes-mediated transformation system was applied for inducing hairy roots. Further, a total of six phenolic compounds and two anthocyanins were analyzed in the hairy roots which were exposed to both light and dark conditions, and their amounts were estimated by HPLC. The gene expression levels peaked on day 5 of culture during the time course of both dark and light conditions. Notably, FtPAL, Ft4CL, FtC4H, FtCHI, FtF3H, FtF3'H-1, and FtFLS-1 were more highly expressed in Hokkai T10 than in Hokkai T8 under dark conditions, among which FtPAL and FtCHI were found to be significantly upregulated, except on day 20 of culture. Significantly higher levels of phenolic compound, rutin, along with two anthocyanins were detected in the hairy roots of Hokkai T10 under both conditions. Furthermore, among all the phenolic compounds detected, the amount of rutin in Hokkai T10 hairy roots was found to be ∼5-fold (59,01 mg/g dry weight) higher than that in the control (12.45 mg/g dry weight) at the respective time periods under light and dark conditions.


Assuntos
Fagopyrum/metabolismo , Flavonoides/biossíntese , Raízes de Plantas/metabolismo , Agrobacterium/genética , Escuridão , Fagopyrum/genética , Regulação da Expressão Gênica de Plantas/genética , Luz , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética , Raízes de Plantas/genética
13.
J Agric Food Chem ; 62(21): 4839-45, 2014 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-24793050

RESUMO

Buckwheat sprouts are a popular food item in many countries. The effects of light-emitting diodes (LEDs) on sprout growth and development, changes in mRNA transcription, and accumulation of phenylpropanoid compounds were studied in tartary buckwheat 'Hokkai T8' sprouts. The highest transcript levels were observed after 2 days of LED exposure for all genes, especially FtPAL and FtF3'H, which showed higher expression in sprouts grown under blue and white light than in those grown under red light. Catechin content in sprouts grown under red light increased dramatically throughout the 10 day time course. Maximum rutin content (43.37 mg/g dry weight (DW)) was observed in sprouts at 4 days after exposure (DAE) to blue light. Similarly, the highest cyanidin 3-O-rutinoside content (0.85 mg/g DW) was detected at 10 DAE to blue light. On the basis of these results, blue LED light is recommended as a light source for enhancing the content of phenolic compounds in tartary buckwheat sprouts.


Assuntos
Fagopyrum/genética , Fagopyrum/efeitos da radiação , Flavonoides/biossíntese , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Proteínas de Plantas/genética , Vias Biossintéticas/efeitos da radiação , Fagopyrum/crescimento & desenvolvimento , Fagopyrum/metabolismo , Alimentos Orgânicos/análise , Luz , Proteínas de Plantas/metabolismo , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Sementes/efeitos da radiação
14.
Nat Prod Commun ; 9(4): 533-7, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24868877

RESUMO

Chinese cabbage is one of the most important leafy vegetables widely used in East Asian cuisines. The glucosinolate (GSL) accumulation and transcript levels of 7 transcription factors (Dof1.1, IQD1-1, MYB28, MYB29, MYB34, MYB51, and MYB122, and their isoforms) involved in the biosynthesis of aliphatic and indolic glucosinolates (GSLs) were analyzed at different stages of Chinese cabbage (Brassica rapa ssp. pekinensis) seedlings under light and dark conditions using high performance liquid chromatography and quantitative real time PCR. During seedling development, transcription of almost all transcription factors under light conditions was higher expressed than under dark conditions. Five aliphatic GSLs (progoitrin, sinigrin, glucoalyssin, gluconapin, and glucobrassicanapin) and four indolic GSLs (4-hydroxyglucobrassicin, glucobrassicin, 4-methoxyglucobrassicin, and neoglucobrasscin) were detected. Total GSL contents under light conditions 6, 8, and 10 days after sowing (DAS) were 3.2-, 3.9-, and 6.9-fold higher, respectively than those of dark conditions. Interestingly, total GSL contents 2 {85.4 micromol/g dry weight (DW)} to 10 (7.74 micromol/g DW) DAS under dark conditions were gradually decreased. In this study, our results suggest that light affects the levels of GSL in Chinese cabbage seedlings. These results could be useful for obtaining cabbage varieties rich in GSLs.


Assuntos
Brassicaceae/metabolismo , Brassicaceae/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Glucosinolatos/metabolismo , Plântula/metabolismo , Fatores de Transcrição/metabolismo , Brassicaceae/genética , Glucosinolatos/química , Luz , Fatores de Transcrição/genética
15.
ScientificWorldJournal ; 2014: 726567, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24605062

RESUMO

Anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR) play an important role in the monomeric units biosynthesis of proanthocyanidins (PAs) such as catechin and epicatechin in several plants. The aim of this study was to clone ANR and LAR genes involved in PAs biosynthesis and examine the expression of these two genes in different organs under different growth conditions in two tartary buckwheat cultivars, Hokkai T8 and T10. Gene expression was carried out by quantitative real-time RT-PCR, and catechin and epicatechin content was analyzed by high performance liquid chromatography. The expression pattern of ANR and LAR did not match the accumulation pattern of PAs in different organs of two cultivars. Epicatechin content was the highest in the flowers of both cultivars and it was affected by light in only Hokkai T8 sprouts. ANR and LAR levels in tartary buckwheat might be regulated by different mechanisms for catechin and epicatechin biosynthesis under light and dark conditions.


Assuntos
Antocianinas/metabolismo , Vias Biossintéticas/genética , Catequina/análise , Fagopyrum/enzimologia , NADH NADPH Oxirredutases/genética , Sequência de Bases , Catequina/biossíntese , Cromatografia Líquida de Alta Pressão , Clonagem Molecular , Biologia Computacional , DNA Complementar/genética , Fagopyrum/química , Dados de Sequência Molecular , Estrutura Molecular , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Especificidade da Espécie
16.
Int J Mol Med ; 33(2): 469-77, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24336971

RESUMO

In this study, we investigated the anti-allergic action of mulberry fruit extract (MFE) or MFE in combination with naringinase (MFEN) in IgE-activated RBL-2H3 cells, and investigated the mechanisms responsible for the anti-allergic effects of MFEN. ß-hexosaminidase release assay was used to measure the amount of ß-hexosaminidase released from the cells, and ELISA was used to measure the levels of tumor necrosis factor-α (TNF-α). We found that MFE significantly reduced the release of ß-hexosaminidase (IC(50), 10.59 mg/ml) and TNF-α (IC(50), 4.87 mg/ml). Moreover, MFEN enhanced the inhibitory effects on the release of ß-hexosaminidase (IC(50), 123.10 µg/ml) and TNF-α (IC(50), 65.01 µg/ml). Furthermore, MFEN had no cytotoxicity at the concentration range used to exert the anti-allergic effects. In addition, we evaluated the effects of MFEN on the formation of pro-inflammatory lipid mediators, such as prostaglandin D(2) (PGD(2)), leukotriene C(4) (LTC(4)) and leukotriene B(4) (LTB(4)) using enzyme immunoassay (EIA) kits. MFEN markedly reduced the formation of PGD(2) (IC(50), 6.47 µg/ml) and LTC(4) (IC(50), 0.31 µg/ml), but not LTB(4) (IC(50), 25.75 µg/ml). In mechanistic analyses, we measured the phosphorylation of Syk, Lyn and Fyn by immunoblot analysis. MFEN significantly inhibited the phosphorylation of Syk, but not that of Lyn or Fyn. MFEN also suppressed the phosphorylation of phospholipase C (PLC)γ1/2, protein kinase C (PKC)δ, linker for activation of T cells (LAT), extracellular signal-regulated protein kinase (ERK)1/2, JNK, GRB2-associated binding protein 2 (Gab2), phosphoinositide-3-kinase (PI3K), Akt, cytosolic phospholipase A2 and 5-lipoxygenase, as well as the expression of cyclooxygenase-2. In conclusion, these results suggest that MFEN exerts potent inhibitory effects on allergic response through the suppression of the activation of the FcεRI signaling cascade. Our data demonstrating the anti-allergic effects of MFEN may provide further insight into the therapeutic application of MFEN or its use as a functional food.


Assuntos
Antialérgicos/farmacologia , Frutas/química , Imunoglobulina E/metabolismo , Morus/química , Complexos Multienzimáticos/metabolismo , Extratos Vegetais/farmacologia , beta-Glucosidase/metabolismo , Animais , Linhagem Celular Tumoral , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Hipersensibilidade/tratamento farmacológico , Concentração Inibidora 50 , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt , Proteínas Proto-Oncogênicas c-fyn/genética , Proteínas Proto-Oncogênicas c-fyn/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais , Quinase Syk , Fator de Necrose Tumoral alfa/metabolismo , Quinases da Família src/genética , Quinases da Família src/metabolismo
17.
Nat Prod Commun ; 8(9): 1233-6, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24273854

RESUMO

Gamma-aminobutyric acid (GABA), synthesized by glutamate decarboxylase (GAD), plays an important role in plants. To study the molecular mechanism of GAD regulation and to examine the levels of GABA in Scutellaria baicalensis, we isolated cDNA clones (SbGAD1 and 2) encoding GAD from S. baicalensis. The open reading frames of SbGADI and 2 were 1,503 and 1,494 bp long and had 450 and 497 amino acid residues, respectively. Quantitative real-time RT-PCR analysis was performed to show the variation of transcript levels among different organs of S. baicalensis. Transcript levels of SbGAD1 and 2 were highest in the root and flower, respectively. The GABA content of different parts (ranked in descending order) was as follows: leaf > flower > stem > root. We concluded that the expression pattern of SbGAD1 and 2 did not match the accumulation pattern of GABA in different organs. We presume that GABA biosynthesis might be more controlled by SbGAD2 than SbGADI. These data will aid in future studies that seek to understand the mechanisms underlying GABA biosynthesis, an important amino acid that is synthesized by the GAD enzyme. To explain adequately the GABA biosynthesis mechanisms in S. baicalensis, the enzyme activities of SbGAD1 and 2 should be determined in the near future.


Assuntos
Glutamato Descarboxilase/genética , Scutellaria baicalensis/genética , Ácido gama-Aminobutírico/biossíntese , Sequência de Bases , Cálcio/metabolismo , Calmodulina/metabolismo , DNA Complementar/química , Glutamato Descarboxilase/metabolismo , Dados de Sequência Molecular , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Scutellaria baicalensis/enzimologia , Análise de Sequência de DNA
18.
J Agric Food Chem ; 61(50): 12356-61, 2013 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-24274859

RESUMO

In this study, the optimum wavelengths of light required for carotenoid biosynthesis were determined by investigating the expression levels of carotenoid biosynthetic genes and carotenoid accumulation in sprouts of tartary buckwheat (Fagopyrum tataricum Gaertn.) exposed to white, blue, and red light-emitting diodes (LEDs). Most carotenoid biosynthetic genes showed higher expression in sprouts irradiated with white light at 8 days after sowing than in those irradiated with blue and red lights. The dominant carotenoids in tartary buckwheat sprouts were lutein and ß-carotene. The richest accumulation of total carotenoids was observed in sprouts grown under white light (1282.63 µg g(-1) dry weight), which was relatively higher than that in sprouts grown under blue and red lights (940.86 and 985.54 µg g(-1), respectively). This study might establish an effective strategy for maximizing the production of carotenoids and other important secondary metabolites in tartary buckwheat sprouts by using LED technology.


Assuntos
Carotenoides/biossíntese , Fagopyrum/efeitos da radiação , Proteínas de Plantas/genética , Vias Biossintéticas/efeitos dos fármacos , Fagopyrum/genética , Fagopyrum/crescimento & desenvolvimento , Fagopyrum/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Luz , Proteínas de Plantas/metabolismo
19.
J Agric Food Chem ; 61(44): 10525-33, 2013 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-24083509

RESUMO

Red-flowered buckwheat ( Fagopyrum esculentum ) is used in the production of tea, juice, and alcohols after the detoxification of fagopyrin. In order to investigate the metabolomics and regulatory of anthocyanin production in red-flowered (Gan-Chao) and white-flowered (Tanno) buckwheat cultivars, quantitative real-time RT-PCR (qRT-PCR), gas chromatography time-of-flight mass spectrometry (GC-TOFMS), and high performance liquid chromatography (HPLC) were conducted. The transcriptions of FePAL, FeC4H, Fe4CL1, FeF3H, FeANS, and FeDFR increased gradually from flowering stage 1 and reached their highest peaks at flowering stage 3 in Gan-Chao flower. In total 44 metabolites, 18 amino acids, 15 organic acids, 7 sugars, 3 sugar alcohols, and 1 amine were detected in Gan-Chao flowers. Two anthocyanins, cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside, were identified in Gan-Chao cultivar. The first component of the partial least-squares to latent structures-discriminate analysis (PLS-DA) indicated that high amounts of phenolic, shikimic, and pyruvic acids were present in Gan-Chao. We suggest that transcriptions of genes involved in anthocyanin biosynthesis, anthocyanin contents, and metabolites have correlation in the red-flowered buckwheat Gan-Chao flowers. Our results may be helpful to understand anthocyanin biosynthesis in red-flowered buckwheat.


Assuntos
Antocianinas/biossíntese , Fagopyrum/química , Fagopyrum/metabolismo , Regulação da Expressão Gênica de Plantas , Metabolômica , Proteínas de Plantas/genética , Antocianinas/química , Cromatografia Líquida de Alta Pressão , Fagopyrum/classificação , Fagopyrum/genética , Espectrometria de Massas , Estrutura Molecular , Proteínas de Plantas/metabolismo
20.
Food Chem Toxicol ; 62: 292-8, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23994088

RESUMO

Previous studies suggested that the hepatoprotective activity of betaine is associated with its effects on sulfur amino acid metabolism. We examined the mechanism by which betaine prevents the progression of alcoholic liver injury and its therapeutic potential. Rats received a liquid ethanol diet for 6 wk. Ethanol consumption elevated serum triglyceride and TNFα levels, alanine aminotransferase and aspartate aminotransferase activities, and lipid accumulation in liver. The oxyradical scavenging capacity of liver was reduced, and expression of CD14, TNFα, COX-2, and iNOS mRNAs was induced markedly. These ethanol-induced changes were all inhibited effectively by betaine supplementation. Hepatic S-adenosylmethionine, cysteine, and glutathione levels, reduced in the ethanol-fed rats, were increased by betaine supplementation. Methionine adenosyltransferase and cystathionine γ-lyase were induced, but cysteine dioxygenase was down-regulated, which appeared to account for the increment in cysteine availability for glutathione synthesis in the rats supplemented with betaine. Betaine supplementation for the final 2 wk of ethanol intake resulted in a similar degree of hepatoprotection, revealing its potential therapeutic value in alcoholic liver. It is concluded that the protective effects of betaine against alcoholic liver injury may be attributed to the fortification of antioxidant defense via improvement of impaired sulfur amino acid metabolism.


Assuntos
Aminoácidos Sulfúricos/metabolismo , Antioxidantes/metabolismo , Betaína/farmacologia , Hepatopatias Alcoólicas/tratamento farmacológico , Animais , Peso Corporal/efeitos dos fármacos , Ciclo-Oxigenase 2/genética , Cisteína/metabolismo , Citosol/efeitos dos fármacos , Citosol/metabolismo , Suplementos Nutricionais , Enzimas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Glutationa/metabolismo , Receptores de Lipopolissacarídeos/genética , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Hepatopatias Alcoólicas/genética , Hepatopatias Alcoólicas/metabolismo , Masculino , Óxido Nítrico Sintase Tipo II/genética , Tamanho do Órgão/efeitos dos fármacos , Ratos Wistar , Fator de Necrose Tumoral alfa/genética
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