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1.
Chin J Integr Med ; 29(2): 186-191, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36527536

RESUMO

Cerebral small vessel disease (CSVD) is a senile brain lesion caused by the abnormal structure and function of arterioles, venules and capillaries in the aging brain. The etiology of CSVD is complex, and disease is often asymptomatic in its early stages. However, as CSVD develops, brain disorders may occur, such as stroke, cognitive dysfunction, dyskinesia and mood disorders, and heart, kidney, eye and systemic disorders. As the population continues to age, the burden of CSVD is increasing. Moreover, there is an urgent need for better screening methods and diagnostic markers for CSVD, in addition to preventive and asymptomatic- and mild-stage treatments. Integrative medicine (IM), which combines the holistic concepts and syndrome differentiations of Chinese medicine with modern medical perspectives, has unique advantages for the prevention and treatment of CSVD. In this review, we summarize the biological markers, ultrasound and imaging features, disease-related genes and risk factors relevant to CSVD diagnosis and screening. Furthermore, we discuss IM-based CSVD prevention and treatment strategies to stimulate further research in this field.


Assuntos
Doenças de Pequenos Vasos Cerebrais , Disfunção Cognitiva , Medicina Integrativa , Acidente Vascular Cerebral , Humanos , Encéfalo/patologia , Doenças de Pequenos Vasos Cerebrais/etiologia , Doenças de Pequenos Vasos Cerebrais/patologia , Acidente Vascular Cerebral/complicações , Disfunção Cognitiva/complicações , Imageamento por Ressonância Magnética
2.
Taiwan J Obstet Gynecol ; 61(2): 299-305, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35361391

RESUMO

OBJECTIVE: During early pregnancy, the proliferation placental cells is crucial for proper implantation and formation of maternal-fetal circulation. Platelet-derived growth factor-AA (PDGF-AA) has been detected in placenta during early pregnancy; however, the role of PDGF-AA in placental cell growth has not been studied extensively. Primary cilium, a centrosome-based cellular protrusion, is an signaling hub for regulating development and differentiation. Importantly, the receptor of PDGF-AA (Pdgfr-α) is detected in the primary cilium and primary cilia-mediated PDGF-AA signaling regulates development and differentiation. Here we would like to investigate whether PDGF-AA regulates placental cell growth and whether primary cilia play roles in this process. MATERIALS AND METHODS: Human placental choriocarcinoma JAR cells were treated with PDGF-AA followed by examining cell growth. Primary cilia and subcellular localization of Pdgfr-α were observed by immunofluorescence staining. Manipulation of primary cilia was performed by treating cells with roscovitine or by transfecting cells with siRNA against IFT88. RESULTS: Here we showed that PDGF-AA induced JAR cell proliferation. In addition, JAR cells grew primary cilia where Pdgfr-α was detected. More importantly, pharmacological inhibition of primary cilia formation or depletion of cilia-related gene, IFT88, alleviated PDGF-AA induced JAR cell proliferation. CONCLUSION: Thus, our study show that PDGF-AA facilitates human placental choriocarcinomaJARcell growth via primary cilia.


Assuntos
Coriocarcinoma , Cílios , Proliferação de Células , Feminino , Humanos , Placenta , Fator de Crescimento Derivado de Plaquetas/farmacologia , Gravidez
3.
Invest Ophthalmol Vis Sci ; 62(13): 12, 2021 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-34661609

RESUMO

Purpose: The purpose of this study was to develop a preclinical compound, ITRI-E-(S)4046, a dual synergistic inhibitor of myosin light chain kinase 4 (MYLK4) and Rho-related protein kinase (ROCK), for reducing intraocular pressure (IOP). Methods: ITRI-E-(S)4046 is an amino-pyrazole derivative with physical and chemical properties suitable for ophthalmic formulation. In vitro kinase inhibition was evaluated using the Kinase-Glo Luminescent Kinase Assays. A comprehensive kinase selectivity analysis of ITRI-E-(S)4046 was performed using the KINOMEscan assay from DiscoverRx. The IOP reduction and tolerability of ITRI-E-(S)4046 were assessed in ocular normotensive rabbits, ocular normotensive non-human primates, and ocular hypertensive rabbits. In vivo studies were conducted to assess drug concentrations in ocular tissue. The adverse ocular effects of rabbit eyes were evaluated following the OECD405 guidelines. Results: ITRI-E-(S)4046 showed highly selective kinase inhibitory activity against ROCK1/2, MYLK4, and mitogen-activated protein kinase kinase kinase 19 (MAP3K19), with high specificity against protein kinase A, G, and C families. In ocular normotensive rabbits and non-human primates, the mean IOP reductions of 0.1% ITRI-E-(S)4046 eye drops were 29.8% and 28.5%, respectively. In hypertonic saline-induced and magnetic beads-induced ocular hypertensive rabbits, the mean IOP reductions of ITRI-E-(S)4046 0.1% eye drops were 46.9% and 22.0%, respectively. ITRI-E-(S)4046 was well tolerated with only temporary and minor signs of hyperemia. Conclusions: ITRI-E-(S)4046 is a novel type of highly specific ROCK1/2 and MYLK4 inhibitor that can reduce IOP in normotensive and hypertensive animal models. It has the potential to become an effective and well-tolerated treatment for glaucoma.


Assuntos
Benzoatos/farmacologia , Proteínas de Ligação ao Cálcio/antagonistas & inibidores , Pressão Intraocular/efeitos dos fármacos , Isoquinolinas/farmacologia , Quinase de Cadeia Leve de Miosina/antagonistas & inibidores , Hipertensão Ocular/tratamento farmacológico , Sulfonamidas/farmacologia , beta-Alanina/análogos & derivados , Animais , Modelos Animais de Doenças , Humanos , Macaca , Masculino , Hipertensão Ocular/fisiopatologia , Coelhos , Tonometria Ocular , beta-Alanina/farmacologia , Quinases Associadas a rho/antagonistas & inibidores
4.
Cells ; 10(6)2021 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-34208028

RESUMO

Etoposide (ETO) has been used in treating adrenocortical tumor (ACT) cells. Our previous study showed that ETO inhibits ACT cell growth. In the present study, we show that ETO treatment at IC50 (10 µM) inhibited ACT cell growth by inducing cellular senescence rather than apoptosis. Several markers of cellular senescence, including enlarged nuclei, activated senescence-associated ß-galactosidase activity, elevated levels of p53 and p21, and down-regulation of Lamin B1, were observed. We further found that ETO induced multiple centrosomes. The inhibition of multiple centrosomes accomplished by treating cells with either roscovitine or centrinone or through the overexpression of NR5A1/SF-1 alleviated ETO-induced senescence, suggesting that ETO triggered senescence via multiple centrosomes. Primary cilia also played a role in ETO-induced senescence. In the mechanism, DNA-PK-Chk2 signaling was activated by ETO treatment; inhibition of this signaling cascade alleviated multiple ETO-induced centrosomes and primary cilia followed by reducing cellular senescence. In addition to DNA damage signaling, autophagy was also triggered by ETO treatment for centrosomal events and senescence. Importantly, the inactivation of DNA-PK-Chk2 signaling reduced ETO-triggered autophagy; however, the inhibition of autophagy did not affect DNA-PK-Chk2 activation. Thus, ETO activated the DNA-PK-Chk2 cascade to facilitate autophagy. The activated autophagy further induced multiple centrosomes and primary cilia followed by triggering senescence.


Assuntos
Neoplasias do Córtex Suprarrenal/patologia , Senescência Celular , Centrossomo/fisiologia , Cílios/efeitos dos fármacos , Etoposídeo/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Neoplasias do Córtex Suprarrenal/tratamento farmacológico , Neoplasias do Córtex Suprarrenal/genética , Neoplasias do Córtex Suprarrenal/metabolismo , Antineoplásicos Fitogênicos/farmacologia , Apoptose , Autofagia , Proliferação de Células , Centrossomo/efeitos dos fármacos , Dano ao DNA , Humanos , Células Tumorais Cultivadas
5.
J Cell Physiol ; 236(4): 2706-2724, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-32869310

RESUMO

Septins play important roles in regulating development and differentiation. Septin 7 (SEPT7) is a crucial component in orchestrating the septin core complex into highly ordered filamentous structures. Here, we showed that genetic depletion of SEPT7 or treatment with forchlorfenuron (FCF; a compound known to affect septin filament assembly) led to reduced the S phase entry in cell models and zebrafish embryos. In addition to colocalizing with actin filaments, SEPT7 resided in the centrosome, and SEPT7 depletion led to aberrant mitotic spindle pole formation. This mitotic defect was rescued in SEPT7-deficient cells by wild-type SEPT7, suggesting that SEPT7 maintained mitotic spindle poles. In addition, we observed disorganized microtubule nucleation and reduced cell migration with SEPT7 depletion. Furthermore, SEPT7 formed a complex with and maintained the abundance of p150glued , the component of centriole subdistal appendages. Depletion of p150glued resulted in a phenotype reminiscent of SEPT7-deficient cells, and overexpression of p150glued reversed the defective phenotypes. Thus, SEPT7 is a centrosomal protein that maintains proper cell proliferation and microtubule array formation via maintaining the abundance of p150glued .


Assuntos
Proteínas de Ciclo Celular/metabolismo , Centrossomo/metabolismo , Complexo Dinactina/metabolismo , Microtúbulos/metabolismo , Fase S , Septinas/metabolismo , Animais , Proteínas de Ciclo Celular/genética , Linhagem Celular Tumoral , Proliferação de Células , Centrossomo/efeitos dos fármacos , Complexo Dinactina/genética , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Microtúbulos/efeitos dos fármacos , Microtúbulos/genética , Compostos de Fenilureia/farmacologia , Piridinas/farmacologia , Fase S/efeitos dos fármacos , Pontos de Checagem da Fase S do Ciclo Celular , Septinas/genética , Transdução de Sinais , Peixe-Zebra/embriologia , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
6.
Int J Mol Sci ; 20(20)2019 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-31640125

RESUMO

Gestational diabetes mellitus (GDM) is a type of unbalanced glucose tolerance that occurs during pregnancy, which affects approximately 10% of pregnancies worldwide. Fetuin-A is associated with insulin resistance, and the concentration of circulating fetuin-A increases in women with GDM, however, the role of fetuin-A in the placenta remains unclear. In this study, we enrolled placental samples from twenty pregnant women with GDM and twenty non-GDM pregnant women and found that the abundance of fetuin-A was upregulated in terms of mRNA and protein levels. Fetuin-A inhibited placental cell growth by inducing apoptosis and inhibiting S phase entry. Irregular alignment of mitotic chromosomes and aberrant mitotic spindle poles were observed. In addition, centrosome amplification was induced by fetuin-A treatment, and these amplified centrosomes nucleated microtubules with disorganized microtubule arrays in placental cells. Furthermore, fetuin-A inhibited autophagy, and thus blocked the growth of the primary cilium, a cellular antenna that regulates placenta development and differentiation. Thus, our study uncovered the novel function of fetuin-A in regulating placental cell growth and ciliogenesis.


Assuntos
Diabetes Gestacional/metabolismo , Placenta/metabolismo , Placentação , alfa-2-Glicoproteína-HS/genética , alfa-2-Glicoproteína-HS/metabolismo , Adulto , Apoptose , Estudos de Casos e Controles , Diferenciação Celular , Células Cultivadas , Centrossomo/metabolismo , Diabetes Gestacional/genética , Feminino , Humanos , Idade Materna , Placenta/citologia , Gravidez , Fuso Acromático/metabolismo , Regulação para Cima
7.
J Cell Physiol ; 234(7): 10445-10457, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30417365

RESUMO

Chloroquine (CQ) is an antimalaria drug that has been used in clinical practice for several decades. One serious complication of CQ treatment is the macular retinopathy caused by the disruption of the retinal pigmented epithelium, leading to vision loss. Little is known about how CQ affects retinal pigmented epithelium. In this study, we found that cell proliferation was reduced by CQ treatment in time and dose-dependent manners. No obvious cell death was detected; however, what was observed instead was G0/G1 arrest during which primary cilium started to grow in the presence of CQ. Pharmacological inhibition of primary cilium formation led to a reduction of cell viability suggesting that CQ-induced primary cilium protected cells from death. In addition to cell growth, with the CQ treatment the retina pigmented epithelium (RPE) cells less flattened with the spindle-like protrusion. When checking the microtubule networks, the microtubule nucleation activity was disrupted in the presence of CQ. The level of p150 glued , the largest subunit of dynactin, was reduced in CQ-treated RPE1 cells, and depletion of p150 glued resulted in a phenotype reminiscent of CQ-treated cells. Thus, CQ treatment reduced the expression of p150 glued , leading to reduced S phase entry and defective microtubule nucleation.


Assuntos
Proliferação de Células/efeitos dos fármacos , Cloroquina/farmacologia , Regulação para Baixo/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Microtúbulos/efeitos dos fármacos , Proteínas Quinases/metabolismo , Retina/efeitos dos fármacos , Animais , Ciclo Celular/efeitos dos fármacos , Linhagem Celular , Linhagem Celular Tumoral , Complexo Dinactina/metabolismo , Células Epiteliais/metabolismo , Humanos , Camundongos , Proteínas Associadas aos Microtúbulos/metabolismo , Microtúbulos/metabolismo , Retina/metabolismo
8.
J Chromatogr A ; 1426: 226-32, 2015 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-26643722

RESUMO

The objective of this study was to establish a practical and reliable analytical method for monitoring trace amounts of Δ(9)-tetrahydrocannabinol (THC) and its metabolites in biological samples. A novel on-line preconcentration capillary electrophoresis method combining large volume sample injection, anion selective exhaustive injection and sweeping was developed to enhance analytical sensitivity. A background buffer composed with 30mM phosphate buffer (pH 2.5) containing 40% methanol and 100mM SDS was used to suppress the electroosmotic flow of the uncoated fused silica capillary (40cm×50µm i.d.). High conductivity buffer (200mM phosphate, pH 2.5) was injected for analyte accumulation. The samples, prepared in phosphate buffer or Tris buffer, were introduced by hydrodynamic injection and electrokinetic injection. After sweeping, the separation was performed in micellar electrokinetic chromatography (MEKC) mode at -15kV. During the method validation, the coefficient of determination of the regression curve was measured at greater than 0.993, and the relative standard deviation and relative error were lower than 11.06% and 9.24%, respectively. Under optimized conditions, an improvement of up to 2000-fold higher sensitivity was achieved. This method was applied to the analysis of urine samples, indicating that it could be satisfactorily utilized in the toxicological and clinical monitoring of cannabis.


Assuntos
Dronabinol/análise , Soluções Tampão , Cromatografia Capilar Eletrocinética Micelar/métodos , Dronabinol/metabolismo , Dronabinol/urina , Eletroforese Capilar , Humanos , Sensibilidade e Especificidade , Detecção do Abuso de Substâncias
9.
Electrophoresis ; 36(19): 2396-403, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26081373

RESUMO

The operating parameters that affect the performance of the online preconcentration technique "analyte focusing by micelle collapse-MEKC (AFMC-MEKC)" were examined using a multivariate approach involving experimental design to determine the sunscreen agents in cosmetics. Compared to the single-variable approach, the advantage of the multivariate approach was that many factors could be investigated simultaneously to obtain the best separation condition. A fractional factorial design was used to identify the fewest significant factors in the central composite design (cCD). The cCD was adopted for evaluating the location of the minimum or maximum response in this study. The influences of the experimental variables on the response were investigated by applying a chromatographic exponential function. The optimized condition and the relationship between the experimental variables were acquired using the JMP software. The ANOVA analysis indicated that the Tris pH value, SDS concentration, and ethanol percentage influenced the separation quality and significantly contributed to the model. The optimized condition of the running buffer was 10 mM Tris buffer (pH 9.5) containing 60 mM SDS, 7 mM γ-CD, and 20% v/v ethanol. The sample was prepared in 100 mM Tris buffer (pH 9.0) containing 7.5 mM SDS and 20% v/v ethanol. The SDS concentration in the sample matrix was slightly greater than the CMC value that makes the micelle be easily collapsed and the analytes be accumulated in the capillary. In addition, sunscreen agents in cosmetics after 1000-fold dilution were successfully determined by AFMC-MEKC.


Assuntos
Cromatografia Capilar Eletrocinética Micelar/métodos , Cosméticos/química , Micelas , Protetores Solares/análise , Análise Multivariada , Protetores Solares/química
10.
J Cell Biochem ; 116(9): 2049-60, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25774020

RESUMO

p150(glued) is the largest subunit of dynactin protein complex, through which cargo vesicles link to the microtubule minus-end directed motor protein dynein. In addition, p150(glued) also locates in the mother centriole where it organizes the subdistal appendage. The components of appendage are dynamically regulated throughout the cell cycle stages, but it is still unclear whether the centrosomal residency of p150(glued) correlated with cell cycle progression. Here we found that p150(glued) was located in the mother centriole during G1/S stage and its centrosomal residency was independent of microtubule transportation. However, the centrosomal p150(glued) became blurred at G2/M phase and this event was not regulated by its phosphorylation. Entering into mitosis, p150(glued) was robustly enriched in the mitotic spindle nearby the spindle poles but not in the centrosome. During serum starvation (G0 stage), p150(glued) appeared at the base of primary cilium and its depletion attenuated starvation-induced primary cilium formation. We also checked its role in the maintenance of centrosome homeostasis and configuration, and found depletion of p150(glued) did not induce centrosome amplification or splitting but inhibited U2OS cell growth. G1 arrest and reduced EdU incorporation were observed in p150(glued) deficient U2OS cells. In addition, cyclin E was downregulated following p150(glued) depletion. The p53/p21 signaling was activated indicating that CDKs were inactivated. The reduced cell growth was ameliorated in the p150(glued) depleted cells when treated with p53 inhibitor. Thus, we have identified the centrosomal targeting of p150(glued) in distinct cell cycle stage and uncovered its role in controlling G1/S transition.


Assuntos
Ciclo Celular , Centrossomo/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Linhagem Celular , Ciclina E/metabolismo , Complexo Dinactina , Humanos , Proteínas Associadas aos Microtúbulos/genética , Microtúbulos/metabolismo , Fosforilação , Fuso Acromático/metabolismo
11.
Exp Ther Med ; 8(4): 1047-1054, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25187795

RESUMO

The aim of the present study was to compare the root canal preparation ability of rotary nickel-titanium (NiTi) Hero 642 and K3 files in curved mandibular or maxillary molars. A total of 40 extracted mandibular molars with two separate mesial canals, an apical width of approximately size ≤15 and a root canal curvature of 15-30° were randomly divided into two groups and instrumented using Hero 642 (n=20) or K3 files (n=20). Canal straightening, working length, transportation, cross-sectional area, minimum dentin thickness and the canal angle curvature degree were examined, and a systematic review of the literature was conducted. No statistically significant differences were observed between the two groups with regard to the mean degree of straightening, mean change in working length, mean transportation, amount of dentin removed or remaining minimum dentin thickness (P>0.05). The canal angle curvature decreased in the two groups postoperatively. The systematic review identified six studies, and overall the two files performed similarly in the majority of categories examined. Therefore, the rotary NiTi Hero 642 and K3 files demonstrated comparable shaping abilities and maintenance of working length.

12.
J Agric Food Chem ; 62(20): 4618-24, 2014 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-24785577

RESUMO

This study aimed to identify the bioactive compounds and evaluate the anti-cold-stress function of the sorghum distillery residue (SDR) using tilapia as an alternative animal model. The highest contents of water-soluble bioactive compounds in SDR were polyphenols, followed by tannins, anthocyanins, and flavonoids. SDR was extracted with double-distilled water, 95% ethanol, and ethyl acetate, separately. The ethanol extract (SDR-E) yielded the highest polyphenol content [15.03 mg/g of SDR dry weight (dw)], of which the EC50 value of R,R-diphenyl-ß-picrylhydrazyl (DPPH) radical scavenging efficiency was 0.56 ± 0.04 mg/mL. The SDR-E suppressed the oxidation of low-density lipoproteins (LDLs) more efficiently than that of other extracts. Tilapia fed a diet containing 3.6% SDR-E decreased accumulative mortality during cold stress, of 46.2%. The accumulative morality of the control was 92.9%. The phenolic acids identified in SDR included gallic acid (0.36 ± 0.08 mg/g of SDR dw), 3,4-dihydroxybenzoic acid (0.16 ± 0.12 mg/g of SDR dw), and 4-hydroxybenzoic acid (0.49 ± 0.23 mg/g of SDR dw). Diets supplemented with 0.5% 4-hydroxybenzoic acid fed to tilapia showed a lower mortality rate than that fed 1.0% 4-hydroxybenzoic acid, comparable to that of the tilapia fed 20% SDR. The latter showed lower mortality than that of the control. These results suggested that 4-hydroxybenzoic acid is one of the major anti-cold-stress compounds in SDR.


Assuntos
Antioxidantes/metabolismo , Hidroxibenzoatos/metabolismo , Extratos Vegetais/metabolismo , Sorghum/química , Tilápia/fisiologia , Resíduos/análise , Ração Animal/análise , Animais , Antioxidantes/química , Temperatura Baixa , Destilação , Hidroxibenzoatos/química , Extratos Vegetais/química , Tilápia/crescimento & desenvolvimento
13.
Mitochondrial DNA ; 25(3): 188-9, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23992243

RESUMO

We determined the complete mitochondrial genome (mitogenome) sequence of Onychostoma alticorpus, which is known as an endemic freshwater species in Taiwan, by using long polymerase chain reaction method. The total length of O. alticorpus mitogenome is 16,680 bp, consisting of 13 protein-coding genes, 22 transfer RNAs, 2 ribosomal RNAs genes and a noncoding control region. The overall base composition of O. alticorpus is 30.88% for A, 23.57% for T, 16.56% for G and 28.99% for C, with a slight AT bias of 54.45%. Gene location and specific usage of distinct termination codon types characterize typically the vertebrate mitochondrial genome. The determination of O. alticorpus mitogenome would play an important role not only in the delineation of phylogeographic history and population genetic structure, but reflection of conservation efforts on the genetic diversity as well as population vitality.


Assuntos
Cyprinidae/genética , Genoma Mitocondrial , Animais , DNA Mitocondrial/genética , Dados de Sequência Molecular , RNA Ribossômico/genética , RNA de Transferência/genética , Análise de Sequência de DNA
14.
Mitochondrial DNA ; 25(5): 377-8, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23992244

RESUMO

The Onychostoma barbatulus is a small cyprinid fish and distributed mainly in Taiwan and mainland China. The total length of O. barbatulus mitogenome is 16,597 bp, and had a gene content consisting of 13 protein-coding genes, 22 transfer RNAs, 2 ribosomal RNAs genes and a noncoding control region. The determination of O. barbatulus mitogenome would play an important role both in the delineation of phylogeographic history and population genetic structure, as well as reflection of conservation efforts on the genetic diversity and population vitality.


Assuntos
Cyprinidae/genética , Genoma Mitocondrial , Análise de Sequência de DNA/métodos , Animais , China , Cyprinidae/classificação , DNA Mitocondrial/análise , Genes Mitocondriais
15.
J Agric Food Chem ; 61(24): 5914-20, 2013 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-23718839

RESUMO

An online stacking capillary electrophoresis (CE) method, cation-selective exhaustive injection sweeping micellar electrokinetic chromatography (CSEI-sweep-MEKC), is developed and optimized for analysis of ractopamine (RP) and its homologue dehydroxyractopamine (DRP) in porcine meat. Chemometric experimental design was used to achieve the best possible optimization and reduce the number of trials and errors. The CSEI-sweep-MEKC method enables nanogram per gram level analysis, with limits of detection (LODs) in meat of 5 ng/g for RP and 3 ng/g for DRP (S/N = 3). A higher conductivity buffer (HCB) zone was injected into the capillary, allowing for the analytes to be electrokinetically injected at a voltage of 9 kV for 12 min. Using 125 mM sodium dodecyl sulfate and 15% methanol in the sweeping buffer, RP and DRP were well-separated. The method was validated with a linear calibration curve of 10-300 ng/g (r > 0.994). In comparison to the normal capillary zone electrophoresis method (1 psi for 10 s), this stacking strategy resulted in 900 times sensitivity enhancement. This technique was further applied for analyzing seven kinds of commercial meats, and the residual RP was detected in one (5.76 ng/g of RP). The data were corresponding to the data analyzed by the commercial testing kit and mass spectrometry spectra. This method was successfully used on real samples and is considered feasible for serving as a tool for routine examination in markets.


Assuntos
Resíduos de Drogas/análise , Contaminação de Alimentos , Inspeção de Alimentos/métodos , Substâncias de Crescimento/análise , Carne/análise , Fenetilaminas/análise , Animais , Cromatografia Capilar Eletrocinética Micelar , Biologia Computacional , Resíduos de Drogas/química , Contaminação de Alimentos/legislação & jurisprudência , Inspeção de Alimentos/legislação & jurisprudência , Inspeção de Alimentos/normas , Rotulagem de Alimentos/normas , Substâncias de Crescimento/química , Hidroxilação , Legislação sobre Alimentos , Limite de Detecção , Carne/economia , Carne/normas , Fenetilaminas/química , Sus scrofa , Taiwan
16.
J Nat Prod ; 76(4): 580-7, 2013 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-23540981

RESUMO

Four new 8,8',7,2'-lignans, (+)-ovafolinin B-9'-O-ß-d-glucopyranoside (1), (-)-ovafolinin B-9'-O-ß-d-glucopyranoside (2), (+)-ovafolinin E-9'-O-ß-d-glucopyranoside (3), and (-)-ovafolinin E-9'-O-ß-d-glucopyranoside (4), two neolignans, eusiderin N (5) and (7S,8R)-3,5,5'-trimethoxy-4',7-epoxy-8,3'-neolignan-9,9'-diol-4-O-ß-d-xylopyranoside (6), and two new chromone glycosides, 5,7-dihydroxy-4H-chromen-4-one-3-O-ß-d-glucopyranoside (7) and 5,7-dihydroxy-4H-chromen-4-one-3-O-ß-d-xylopyranoside (8), together with 25 known compounds, were isolated from the stems of Eurya japonica. Structural elucidation of compounds 1-8 was established by spectroscopic methods, especially 2D NMR techniques, electronic circular dichroism data, and comparison with reported data. The isolates were evaluated for antioxidant and anti-NO production activities. Compounds 1, 2, 12-20, and 29 (ED50 23.40 µM for 1) demonstrated potent antioxidant activity compared to the positive control α-tocopherol (ED50 27.21 µM). On the other hand, compounds 1, 2, 7-9, 12-20, and 32 showed only weak anti-NO production activity when compared to the positive control quercetin.


Assuntos
Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Cromonas/isolamento & purificação , Cromonas/farmacologia , Glicosídeos/isolamento & purificação , Glicosídeos/farmacologia , Lignanas/isolamento & purificação , Lignanas/farmacologia , Theaceae/química , Antioxidantes/química , Compostos de Bifenilo/farmacologia , Cromonas/química , Glicosídeos/química , Lignanas/química , Estrutura Molecular , Óxido Nítrico/biossíntese , Ressonância Magnética Nuclear Biomolecular , Picratos/farmacologia , Caules de Planta/química , Quercetina/farmacologia , Estereoisomerismo , Taiwan , alfa-Tocoferol/farmacologia
17.
J Agric Food Chem ; 61(12): 2979-86, 2013 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-23432055

RESUMO

This paper reports that bioassay-guided fractionations of EtOH extract of Momordica charantia fruits led to the isolation of 15 cucurbitane-type triterpene glycosides including 4 new compounds, kuguaosides A-D (1-4), along with 11 known ones, charantoside A (5), momordicosides I (6), F1 (7), F2 (8), K (9), L (10), and U (11), goyaglycosides-b (12) and -d (13), 7ß,25-dihydroxycucurbita-5,23(E)-dien-19-al 3-O-ß-d-allopyranoside (14), and 25-hydroxy-5ß,19-epoxycucurbita-6,23-dien-19-on-3ß-ol 3-O-ß-d-glucopyranoside (15). Their structures were elucidated on the basis of spectroscopic analyses and chemical methods. This study also established the HPLC-ELSD fingerprinting profile of an antiproliferative fraction of which 11 main peaks were identified. Biological evaluation showed that several isolated cucurbitane-type triterpene glycosides had antiproliferative activities against MCF-7, WiDr, HEp-2, and Doay human tumor cell lines. In addition, compound 14 showed potent hypoglycemic activities by glucose uptake assay.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Frutas/química , Glicosídeos/farmacologia , Hipoglicemiantes/farmacologia , Momordica charantia/química , Triterpenos/farmacologia , Animais , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Glicosídeos/química , Glicosídeos/isolamento & purificação , Humanos , Camundongos , Estrutura Molecular , Extratos Vegetais/química , Triterpenos/química , Triterpenos/isolamento & purificação
18.
Anal Bioanal Chem ; 405(6): 1969-76, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23263519

RESUMO

This study describes an on-line stacking CE approach by sweeping with whole capillary sample filling for analyzing five anabolic androgenic steroids in urine samples. The five anabolic steroids for detection were androstenedione, testosterone, epitestosterone, boldenone, and clostebol. Anabolic androgenic steroids are abused in sport doping because they can promote muscle growth. Therefore, a sensitive detection method is imperatively required for monitoring the urine samples of athletes. In this research, an interesting and reliable stacking capillary electrophoresis method was established for analysis of anabolic steroids in urine. After liquid-liquid extraction by n-hexane, the supernatant was dried and reconstituted with 30 mM phosphate buffer (pH 5.00) and loaded into the capillary by hydrodynamic injection (10 psi, 99.9 s). The stacking and separation were simultaneously accomplished at -20 kV in phosphate buffer (30 mM, pH 5.0) containing 100 mM sodium dodecyl sulfate and 40 % methanol. During the method validation, calibration curves were linear (r≥0.990) over a range of 50-1,000 ng/mL for the five analytes. In the evaluation of precision and accuracy for this method, the absolute values of the RSD and the RE in the intra-day (n=3) and inter-day (n=5) analyses were all less than 6.6 %. The limit of detection for the five analytes was 30 ng/mL (S/N=5, sampling 99.9 s at 10 psi). Compared with simple MECK, this stacking method possessed a 108- to 175-fold increase in sensitivity. This simple and sensitive stacking method could be used as a powerful tool for monitoring the illegal use of doping.


Assuntos
Androstenodiona/urina , Eletroforese Capilar/métodos , Epitestosterona/urina , Detecção do Abuso de Substâncias/métodos , Testosterona/análogos & derivados , Testosterona/urina , Atletas , Calibragem , Dopagem Esportivo/prevenção & controle , Análise de Injeção de Fluxo , Hexanos/química , Humanos , Extração Líquido-Líquido , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
19.
Mitochondrial DNA ; 24(2): 108-10, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23025525

RESUMO

In this study, we sequenced the complete mitochondrial genome of Scaphiodonichthys acanthopterus (Cypriniformes, Cyprinidae). This mitochondrial genome, consisting of 16,612 base pairs (bp), encoded genes for 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, and one non-coding control region as those found in other vertebrates, with the gene order identical to that of typical vertebrates. The control region, of 941 bp in length, is located between tRNA (Pro) and tRNA (Phe) . The overall base composition of the heavy strand shows T 25.23%, C 26.85%, A 31.96%, and G 15.95%, with a slight AT bias of 57.19%.


Assuntos
Cyprinidae/genética , DNA Mitocondrial/genética , Animais , RNA Ribossômico/genética , RNA de Transferência/genética
20.
J Tradit Complement Med ; 3(1): 33-40, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24716153

RESUMO

Seventeen compounds, quercetin-3-O-α-l-rhamnoside (1), kaempferol-3-O-α-L-rhamnoside (2), apigenin-7-O-ß-D-glucuronide (3), apigenin 7-O-ß-D-glucuronide methyl ester (4), apigenin 7-O-ß-D-glucuronide ethyl ester (5), chrysoeriol (6), apigenin (7), kaempferol (8), luteolin (9), quercetin (10), methyl 3,4-dihydroxybenzoate (11), p-coumaric acid (12), 4-hydroxybenzoic acid (13), hydroquinone (14), protocathehuic acid (15), gallic acid (16), and indole-3-carboxylic acid (17), were isolated from the ethanol extract of Taiwanese Cardiospermum halicabum. All chemical structures were determined by physical and extensive spectroscopic analyses such as (1) H Nuclear Magnetic Resonance spectroscopy (NMR), (13)C NMR, (1)H-(1)H Correlation spectroscopy ((1)H-(1)H COSY), Heteronuclear Multiple Quantum Coherence spectroscopy (HMQC), Heteronuclear Multiple-bond Correlation spectroscopy (HMBC), and Nuclear Overhauser Effect spectroscopy (NOESY), as well as comparison with literature values. Furthermore, the High-Performance Liquid Chromatography- Photodiode Array Detector (HPLC-DAD) fingerprint profile was established for the determination of major constituents in the EtOAc extract and retention times of the isolated compounds. All isolated compounds were also evaluated for antiinflammatory and antioxidant activities.

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