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1.
Curr Res Food Sci ; 7: 100647, 2023.
Article in English | MEDLINE | ID: mdl-38077468

ABSTRACT

Consumption of aflatoxin-contaminated food can cause severe illness when consumed by humans or livestock. Because the mycotoxin frequently occurs in cereal grains and other agricultural crops, it is crucial to develop portable devices that can be used non-destructively and in real-time to identify aflatoxin-contaminated food materials during early stages of harvesting or processing. In this study, an aflatoxin detection method was developed using a compact Raman device that can be used in the field. Data were obtained using maize samples naturally contaminated with aflatoxin, and the data were analyzed using a machine learning method. Of the multiple classification models evaluated, such as linear discriminant analysis (LDA), linear support vector machines (LSVM), quadratic discriminant analysis (QDA), and quadratic support vector machines and spectral preprocessing methods, the best classification accuracy was achieved at 95.7% using LDA in combination with Savitzky-Golay 2nd derivative (SG2) preprocessing. Partial least squares regression (PLSR) models demonstrated a close-range accuracy within the scope of standard normal variate (SNV) and multiplicative scatter correction (MSC) preprocessing methods, with determination of coefficient values of R2C and R2V of 0.9998 and 0.8322 respectively for SNV, and 0.9916 and 0.8387 respectively for MSC. This study demonstrates the potential use of compact and automated Raman spectroscopy, coupled with chemometrics and machine learning methods, as a tool for rapidly screening food and feed for hazardous substances at on-site field processing locations.

2.
Molecules ; 28(14)2023 Jul 22.
Article in English | MEDLINE | ID: mdl-37513461

ABSTRACT

Pesticides effectively reduce the population of various pests that harm crops and increase productivity, but leave residues that adversely affect health and the environment. Here, a simultaneous multicomponent analysis method based on ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) pretreated by the QuEChERS method was developed to control the maximum residual levels. Among the 140 pesticides with high frequency of detection in agricultural products in Gyeongnam region in Korea for 5 years, 12 pesticides with high detection frequency in sweet pepper were selected. The analytical method is validated, linearities are r2 > 0.999, limit of detection (LOD) ranges from 1.4 to 3.2 µg/kg, and limit of quantification (LOQ) ranges from 4.1 to 9.7 µg/kg, and the recovery rate was 81.7-99.7%. In addition, it was confirmed that a meaningful value of these parameters can be achieved by determining the measurement uncertainty. The results proved that parameters such as recovery rate and relative standard deviation of the analysis method were within international standards. Using the developed method, better and safer sweet peppers will be provided to consumers, and effective pesticide residue management will be possible by expanding to other agricultural products.


Subject(s)
Capsicum , Pesticide Residues , Pesticides , Chromatography, High Pressure Liquid/methods , Pesticides/analysis , Mass Spectrometry/methods , Pesticide Residues/analysis
3.
Toxins (Basel) ; 15(7)2023 07 22.
Article in English | MEDLINE | ID: mdl-37505741

ABSTRACT

Aflatoxins and fumonisins, commonly found in maize and maize-derived products, frequently co-occur and can cause dangerous illness in humans and animals if ingested in large amounts. Efforts are being made to develop suitable analytical methods for screening that can rapidly detect mycotoxins in order to prevent illness through early detection. A method for classifying contaminated maize by applying hyperspectral imaging techniques including reflectance in the visible and near-infrared (VNIR) and short-wave infrared (SWIR) regions, and fluorescence was investigated. Machine learning classification models in combination with different preprocessing methods were applied to screen ground maize samples for naturally occurring aflatoxin and fumonisin as single contaminants and as co-contaminants. Partial least squares-discriminant analysis (PLS-DA) and support vector machine (SVM) with the radial basis function (RBF) kernel were employed as classification models using cut-off values of each mycotoxin. The classification performance of the SVM was better than that of PLS-DA, and the highest classification accuracies for fluorescence, VNIR, and SWIR were 89.1%, 71.7%, and 95.7%, respectively. SWIR imaging with the SVM model resulted in higher classification accuracies compared to the fluorescence and VNIR models, suggesting that as an alternative to conventional wet chemical methods, the hyperspectral SWIR imaging detection model may be the more effective and efficient analytical tool for mycotoxin analysis compared to fluorescence or VNIR imaging models. These methods represent a food safety screening tool capable of rapidly detecting mycotoxins in maize or other food ingredients consumed by animals or humans.


Subject(s)
Aflatoxins , Fumonisins , Mycotoxins , Humans , Animals , Aflatoxins/analysis , Fumonisins/analysis , Zea mays , Hyperspectral Imaging
4.
Curr Res Food Sci ; 6: 100532, 2023.
Article in English | MEDLINE | ID: mdl-37377492

ABSTRACT

This study presents a method for discriminating the geographical origin of dried chili peppers using femtosecond laser ablation-inductively coupled plasma-mass spectrometry (fsLA-ICP-MS) and multivariate analysis, such as orthogonal partial least squares discriminant analysis (OPLS-DA), heatmap analysis, and canonical discriminant analysis (CDA). Herein, 102 samples were analyzed for the content of 33 elements using optimized conditions of 200 Hz (repetition rate), 50 µm (spot size), and 90% (energy). Significant differences in count per second (cps) values of the elements were observed between domestic and imported peppers, with variations of up to 5.66 times (133Cs). The OPLS-DA model accuracy achieved an R2 of 0.811 and a Q2 of 0.733 for distinguishing dried chili peppers of different geographical origins. The variable importance in projection (VIP) and s-plot identified elements 10 and 3 as key to the OPLS-DA model, and in the heatmap, six elements were estimated to be significant in discriminating between domestic and imported samples. Furthermore, CDA showed a high accuracy of 99.02%. This method can ensure food safety for consumers, and accurately determine the geographic origin of agricultural products.

5.
Food Chem ; 427: 136675, 2023 Nov 30.
Article in English | MEDLINE | ID: mdl-37385061

ABSTRACT

In this study, we developed a method for detecting 335 pesticides in ginseng using liquid chromatography quadrupole mass spectrometry (LC-MS/MS) and gas chromatography quadrupole mass spectrometry (GC-MS/MS). Additionally, the linearity, sensitivity, selectivity, accuracy, and precision of the method was validated. The limits of detection (LOD) and limits of quantification (LOQ) for the instrument used in these experiments was 0.1-5.8 µg/kg and 0.3-17.5 µg/kg, respectively. The average recovery was 71.6-113.4%. From 2016 to 2019, 467 ginseng samples were analyzed, of which 304 samples detected pesticide residues, but most of them were below the standard. It can be observed that the hazard quotient (HQ) of ginseng for detected pesticides was less than 1, thus implying that the risk was low. Hence, in this study, we developed a specific, reliable, and suitable method for a fast and simultaneous analysis of 335 pesticides in ginseng.


Subject(s)
Panax , Pesticide Residues , Pesticides , Pesticides/analysis , Tandem Mass Spectrometry/methods , Gas Chromatography-Mass Spectrometry/methods , Chromatography, Liquid/methods , Panax/chemistry , Pesticide Residues/analysis , Risk Assessment
6.
Food Chem ; 423: 136235, 2023 Oct 15.
Article in English | MEDLINE | ID: mdl-37163917

ABSTRACT

Kimchi has been designated as one of the world's five healthiest foods, and it is a traditional Korean fermented food. The purpose of this study was to investigate whether the origin of Kimchi can be discriminated against by using inorganic elements to develop a more accurate method. The OPLS-DA showed that the R2 and Q2 values were 0.908 and 0.81, a high-quality model. We selected 24 elements (133Cs, 238U, 88Sr, K, 157Gd, Na, Mg, 139La, P, 141Pr, 72Ge, 146Nd, 147Sm, 153Eu, 55Mn, 165Ho, 163Dy, 166Er, Fe, 172Yb, 169Tm, 185Re, 175Lu, and 118Sn) with VIP 1 or higher in the OPLS-DA model. In ROC, the selected elements had an accuracy of 100%. The heatmap confirmed the contents of rare earth elements (REEs) in Korean and Chinese Kimchi, the accuracy of distinguishing classification in CDA is 100%. Such results will help to distinguish the origin of agricultural products through inorganic analysis.


Subject(s)
Fermented Foods , Metals, Rare Earth , Mass Spectrometry/methods , Metals, Rare Earth/analysis , Multivariate Analysis , Spectrum Analysis
7.
J Food Sci ; 87(11): 5099-5106, 2022 Nov.
Article in English | MEDLINE | ID: mdl-36201398

ABSTRACT

Lactic acid bacteria (LAB) are not only the most common probiotics in the food and feed industry but are also used as plant probiotics. Therefore, precise identification of LAB at the species level is required. In this study, we compared three different methods, the VITEK 2 ANC card, species-specific PCR, and MALDI-TOF MS, to identify six LAB (Lacticaseibacillus casei, Lacticaseibacillus paracasei, Lacticaseibacillus rhamnosus, Lactiplantibacillus plantarum, Lentilactobacillus buchneri, and Limosilactobacillus fermentum) species previously assigned to the genus Lactobacillus that are used as biofertilizers. Twenty-two strains of six LAB species were analyzed using the VITEK 2 ANC card, species-specific PCR, and MALDI-TOF MS, and identification rates at the species level were 45.5%, 95.5%, and 95.5%, respectively. There were cross-reactions between L. casei and L. parpacasei, and one strain of L. casei could not be identified by these three methods. PCR assays and MALDI-TOF MS were applicable for LAB identification. PRACTICAL APPLICATION: LAB are the most common probiotics in the food and feed industry, so precise identification and classification of LAB at the species level are required. This study aimed at comparing three different methods for the effective identification of six LAB species: biochemical testing using VITEK 2 ANC card, species-specific PCR, and MALDI-TOF MS analysis.


Subject(s)
Lacticaseibacillus casei , Lactobacillales , Lactobacillales/genetics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Polymerase Chain Reaction/methods , Lactobacillaceae , Lacticaseibacillus casei/genetics
8.
3 Biotech ; 11(7): 337, 2021 Jul.
Article in English | MEDLINE | ID: mdl-34221808

ABSTRACT

2-C-Methyl-d-erythrol-4-phosphate (MEP) pathway in plant supplies isoprene building blocks for carotenoids and chlorophylls essential in photosynthesis as well as plant hormones such as gibberellin and abscisic acid. To assess the effect of overexpression of the terminal enzyme of the MEP pathway, 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (HDR), transgenic Nicotiana tabacum overexpressing class 2 HDR from Ginkgo biloba (GbHDR2) under the control of 35S promoter was constructed. Contents of chlorophylls a and b in transgenic tobacco were enhanced by 19 and 7%, respectively, compared to those of the wild type. The carotenoid level was also 18% higher than that in the control plant. As a result, photosynthetic rate of the transgenic tobacco was increased by up to 51%. Diterepenoid duvatrienediol content of transgenic tobacco was also elevated by at least sixfold. To explore the molecular basis of the enhanced isoprenoid accumulation, transcript levels of the key genes involved in the isoprenoid biosynthesis were measured. Transcript levels of geranylgeranyl diphosphate synthase (GGPP), kaurene synthase (KS), gibberellic acid 20 oxidase (GA20ox), and phytoene desaturase (PD) genes in the transgenic tobacco leaves were about twofold higher compared to the wild type. Therefore, upregulation of down-stream genes involved in biosynthesis of di- and tetraterpenoids due to GbHDR2 overexpression was responsible for elevated production of isoprenoids and enhanced photosynthetic rate. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02887-5.

9.
ACS Omega ; 6(19): 12820-12830, 2021 May 18.
Article in English | MEDLINE | ID: mdl-34056433

ABSTRACT

Platycodon grandiflorum is a perennial plant that has been used for medicinal purposes. Specifically, it is widely used in Northern China and Korea for the treatment of various diseases. Terpenoids belong to a group called secondary metabolites and have attracted a wide range of interest. Here, we determined the expressed sequence tag (EST) library of the methyl jasmonate (MeJA)-treated hairy root of P. grandiflorum. In total, 5760 ESTs were obtained, but after deleting the vector sequences and removing poor-quality sequences, a total of 2536 ESTs were attained. Of these, 811 contigs and 1725 singletons were annotated. The data were further analyzed with a focus on the gene families of the terpenoid biosynthetic pathway (TBP). We identified and characterized four TBP genes; among these were three full-length cDNAs encoding PgHMGS, PgMK, and PgMVD, whereas PgHMGR had a partial sequence based on the EST library database. Phylogenetic analysis and a pairwise identity matrix showed that these identified genes were closely related to those of other higher plants. Moreover, the tertiary structure and multiple alignment analysis showed that several distinct conserved motifs were present. Quantitative reverse transcription-polymerase chain reaction results revealed that TBP genes were constitutively expressed in all organs of P. grandiflorum, while the expression of transcript levels of these genes varied distinctly among the organs. Additionally, the total amount of platycosides was highly detected in the root, accumulating in concentrations 7.8 and 2.6 times higher than in the hairy root and stem, respectively, and 1.4 times higher than in the leaf and flower. The highest amount of total phytosterols was found to accumulate in the leaves at 9.3, 9.1, 1.8, and 1.6 times higher than that of the stem, root, hairy root, and flower, respectively. After the hairy root was exposed to the MeJA treatment, the transcript levels of PgHMGS, PgHMGR, PgMK, and PgMVD had significantly increased. The highest level of transcript accumulation occurred at 3 h after initial exposure for most of the genes. Meanwhile, triterpene saponin accumulation increased with the increase in the time of exposure, and at 48 h after initial exposure, the total saponin content was the highest recorded.

10.
ACS Synth Biol ; 3(10): 773-9, 2014 Oct 17.
Article in English | MEDLINE | ID: mdl-24933610

ABSTRACT

To elucidate the function of mevalonate-5-pyrophosphate decarboxylase (MVD) and farnesyl pyrophosphate synthase (FPS) in triterpene biosynthesis, the genes governing the expression of these enzymes were transformed into Panax ginseng hairy roots. All the transgenic lines showed higher expression levels of PgMVD and PgFPS than that by the wild-type control. Among the hairy root lines transformed with PgMVD, M18 showed the highest level of transcription compared to the control (14.5-fold higher). Transcriptions of F11 and F20 transformed with PgFPS showed 11.1-fold higher level compared with control. In triterpene analysis, M25 of PgMVD produced 4.4-fold higher stigmasterol content (138.95 µg/100 mg, dry weight [DW]) than that by the control; F17 of PgFPS showed the highest total ginsenoside (36.42 mg/g DW) content, which was 2.4-fold higher compared with control. Our results indicate that metabolic engineering in P. ginseng was successfully achieved through Agrobacterium rhizogenes-mediated transformation and that the accumulation of phytosterols and ginsenosides was enhanced by introducing the PgMVD and PgFPS genes into the hairy roots of the plant. Our results suggest that PgMVD and PgFPS play an important role in the triterpene biosynthesis of P. ginseng.


Subject(s)
Carboxy-Lyases/metabolism , Geranyltranstransferase/metabolism , Panax/metabolism , Triterpenes/metabolism , Carboxy-Lyases/genetics , Gas Chromatography-Mass Spectrometry , Geranyltranstransferase/genetics , Metabolic Engineering , Panax/genetics , Plant Roots/metabolism , Plants, Genetically Modified , Synthetic Biology , Triterpenes/chemistry , Up-Regulation
11.
J Agric Food Chem ; 61(8): 1928-34, 2013 Feb 27.
Article in English | MEDLINE | ID: mdl-23298228

ABSTRACT

3-Hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) catalyzes the rate-limiting step in the mevalonate pathway. To elucidate the functions of HMGR in triterpene biosynthesis, Platycodon grandiflorum was transformed with a construct expressing Panax ginseng HMGR (PgHMGR). We used PCR analysis to select transformed hairy root lines and selected six lines for further investigation. Quantitative real-time PCR showed higher expression levels of HMGR and total platycoside levels (1.5-2.5-fold increase) in transgenic lines than in controls. Phytosterols levels were also 1.1-1.6-fold higher in transgenic lines than in controls. Among these lines, line T7 produced the highest level of total platycosides (1.60 ± 0.2 mg g(-1) dry weight) and α-spinasterol (1.78 ± 0.16 mg g(-1) dry weight). These results suggest that metabolic engineering of P. grandiflorum by Agrobacterium-mediated genetic transformation may enhance production of phytosterols and triterpenoids.


Subject(s)
Acyl Coenzyme A/genetics , Campanulaceae/metabolism , Panax/enzymology , Phytosterols/metabolism , Plant Proteins/genetics , Plant Roots/metabolism , Plants, Genetically Modified/metabolism , Triterpenes/metabolism , Acyl Coenzyme A/metabolism , Campanulaceae/genetics , Cells, Cultured , Gene Expression , Panax/genetics , Plant Proteins/metabolism , Plant Roots/cytology , Plants, Genetically Modified/genetics
12.
Clin Cancer Res ; 18(11): 3071-8, 2012 Jun 01.
Article in English | MEDLINE | ID: mdl-22472175

ABSTRACT

PURPOSE: Anaplastic thyroid carcinoma (ATC) is one of the most invasive human cancers and has a poor prognosis. Molecular targets of ATC that determine its highly aggressive nature remain unidentified. This study investigated L1 cell adhesion molecule (L1CAM) expression and its role in tumorigenesis of ATCs. EXPERIMENTAL DESIGN: Expression of L1CAM in thyroid cancer was evaluated by immunohistochemical analyses of tumor samples from patients with thyroid cancer. We investigated the role of L1CAM in proliferation, migration, invasion, and chemoresistance using short hairpin RNA (shRNA) knockdown experiments in human ATC cell lines. Finally, we evaluated the role of L1CAM on tumorigenesis with ATC xenograft assay in a nude mouse model. RESULTS: L1CAM expression was not detectable in normal follicular epithelial cells of the thyroid or in differentiated thyroid carcinoma. In contrast, analysis of ATC samples showed specifically higher expression of L1CAM in the invasive area of the tumor. Specific knockdown of L1CAM in the ATC cell lines, FRO and 8505C, caused a significant decrease in the proliferative, migratory, and invasive capabilities of the cells. Suppression of L1CAM expression in ATC cell lines increased chemosensitivity to gemcitabine or paclitaxel. Finally, in an ATC xenograft model, depletion of L1CAM markedly reduced tumor growth and increased the survival of tumor-bearing mice. CONCLUSIONS: We report that L1CAM is highly expressed in the samples taken from patients with ATCs. L1CAM plays an important role in determining tumor behavior and chemosensitivity in cell lines derived from ATCs. Therefore, we suggest that L1CAM may be an important therapeutic target in patients with ATCs.


Subject(s)
Neural Cell Adhesion Molecule L1/metabolism , Thyroid Neoplasms/metabolism , Animals , Biomarkers, Pharmacological , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Knockdown Techniques , Humans , Mice , Mice, Nude , Neoplasm Invasiveness , Neoplasm Transplantation , Neural Cell Adhesion Molecule L1/genetics , RNA, Small Interfering , Thyroid Carcinoma, Anaplastic , Transplantation, Heterologous
13.
J Ginseng Res ; 36(2): 153-60, 2012 Apr.
Article in English | MEDLINE | ID: mdl-23717115

ABSTRACT

Panax ginseng has long been used as a traditional herbal medicine. More recently, it has received attention for its anti-diabetic and anti-obesity effects in humans and in animal models of type 2 diabetes. In the present study, we tested the hypoglycemic effects of ginseng berry extract in beta-cell-deficient mice and investigated the mechanisms involved. Red (ripe) and green (unripe) berry extracts were prepared and administered orally (100 or 200 mg/kg body weight) to streptozotocin-induced diabetic mice daily for 10 wk. The body weight was measured daily, and the nonfasting blood glucose levels were measured after 5 and 10 wk after administration. Glucose tolerance tests were performed, and the serum insulin levels were measured. The proliferation of betacells was measured in vitro. The administration of red or green ginseng berry extract significantly reduced the blood glucose levels and improved the glucose tolerance in beta-cell deficient mice, with the higher doses resulting in better effects. Glucose-stimulated insulin secretion was significantly increased in berry extract-treated mice compared with streptozotocin-induced diabetic control mice. Treatment with ginseng berry extract increased beta-cell proliferation in vitro. Both red berry and green berry extracts improved glycemic control in streptozotocin-induced diabetic mice and increased insulin secretion, possibly due to increased beta-cell proliferation. These results suggest that ginseng berry extracts might have beneficial effects on beta-cell regeneration.

14.
J Agric Food Chem ; 59(24): 12948-53, 2011 Dec 28.
Article in English | MEDLINE | ID: mdl-22087851

ABSTRACT

Sorgoleone is a major component of the hydrophobic root exudate of Sorghum bicolor and is of particular interest to plant chemical ecology as well as agriculture. Sorgoleone was evaluated in this study to observe the expression levels of genes involved in its biosynthesis in response to auxins. Sorgoleone content varied widely according to the duration of application and the concentrations of the auxins. When the application time was increased, the sorgoleone content increased accordingly for all concentrations of IBA (1, 3, and 5 mg/L) and at 1 mg/L for both IAA and NAA. In this study, five different sorgoleone biosynthetic genes were observed, namely DES2, DES3, ARS1, ARS2, and OMT3, which are upregulated in response to IAA, IBA, and NAA. Transcript accumulation was apparent for all genes, but particularly for DES2, which increased up to 475-fold and 180-fold following 72 h exposure to NAA and IBA, respectively, compared to no treatment.


Subject(s)
Gene Expression/drug effects , Indoleacetic Acids/pharmacology , Lipids/biosynthesis , Lipids/genetics , Plant Roots/genetics , Sorghum/genetics , Benzoquinones , Herbicides , Plant Roots/drug effects , Plant Roots/metabolism , Sorghum/drug effects , Sorghum/metabolism
15.
J Clin Endocrinol Metab ; 96(1): E19-30, 2011 Jan.
Article in English | MEDLINE | ID: mdl-20926530

ABSTRACT

CONTEXT: The oncogenic BRAF(V600E) mutation results in an active structural conformation characterized by greatly elevated ERK activity. However, additional cellular effects caused by subcellular action of BRAF(V600E) remain to be identified. OBJECTIVE: To explore these effects, differences in the subcellular localization of wild-type and mutant BRAF in thyroid cancer were investigated. RESULTS: A significant proportion of endogenous and exogenous BRAF(V600E), but not wild-type BRAF, was detected in the mitochondrial fraction, similar to other BRAF mutants including BRAF(V600D), BRAF(V600K), BRAF(V600R), and BRAF(G469A), which showed elevated kinase activity and mitochondrial localization. Induced expression of BRAF(V600E) suppressed the apoptotic responses against staurosporine and TNFα/cycloheximide. Interestingly, the mitochondrial localization and antiapoptotic activities of BRAF(V600E) were unaffected by sorafenib and U0126 suppression of MAPK kinase (MEK) and ERK activities. Similarly, although the RAF inhibitor sorafenib effectively inhibited MEK/ERK activation, it did not block the mitochondrial localization of BRAF(V600E). In addition, inducible expression of BRAF(V600E) increased the glucose uptake rate and decreased O(2) consumption, suggesting that BRAF(V600E) reduces mitochondrial oxidative phosphorylation, a signature feature of cancer cells. Again, these metabolic alterations resulted by BRAF(V600E) expression were not affected by the treatment of thyroid cells by sorafenib. Therefore, RAF and MEK inhibitors are unable to block the antiapoptotic activity of BRAF(V600E) or correct the high glucose uptake rate and glycolytic activity and suppressed mitochondrial oxidative phosphorylation induced by BRAF(V600E). CONCLUSIONS: The mitochondrial localization observed in oncogenic BRAF mutants might be related to their altered responses to apoptotic stimuli and characteristic metabolic phenotypes found in thyroid cancer. The inability of MEK and RAF inhibitors, U0126 and sorafenib, respectively, to block the mitochondrial localization of BRAF(V600E) has additional therapeutic implications for BRAF(V600E)-positive thyroid cancers.


Subject(s)
Benzenesulfonates/pharmacology , Mitochondria/genetics , Proto-Oncogene Proteins B-raf/genetics , Pyridines/pharmacology , Thyroid Gland/metabolism , Thyroid Neoplasms/genetics , Animals , Apoptosis/genetics , Blotting, Western , Cell Line, Tumor , Cell Proliferation/drug effects , Cells, Cultured , Extracellular Signal-Regulated MAP Kinases/genetics , Extracellular Signal-Regulated MAP Kinases/metabolism , Fluorescent Antibody Technique , Humans , Mice , Mice, Transgenic , Mitochondria/drug effects , Mitochondria/metabolism , Mitogen-Activated Protein Kinase Kinases/genetics , Mitogen-Activated Protein Kinase Kinases/metabolism , Mutation , Niacinamide/analogs & derivatives , Phenylurea Compounds , Phosphorylation/drug effects , Protein Kinase Inhibitors/pharmacology , Proto-Oncogene Proteins B-raf/metabolism , Sorafenib , Subcellular Fractions/drug effects , Subcellular Fractions/metabolism , Thyroid Neoplasms/metabolism
16.
Mol Biol Rep ; 38(4): 2307-13, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21052843

ABSTRACT

The balloon flower (Platycodon grandiflorum) is a popular traditional medicinal plant used in Korea to treat conditions such as bronchitis, asthma, tuberculosis, diabetes, and inflammatory diseases. Recently, immunopharmacological research identified triterpenoid and saponin as important active compounds in P. grandiflorum. To study and extract these compounds and other metabolites from P. grandiflorum, a technique was developed for producing hairy root cultures, which are a reliable source of plant compounds. To achieve this, the activity of Agrobacterium rhizogenes was exploited, which can transfer DNA segments into plant genomes after infecting them. In this study, the A. rhizogenes strain R1000 was determined that had the highest infection frequency (87.5%) and induced the most hairy roots per plant, and the concentration of antibiotics (75 mg/l kanamycin) was elucidated for selection after transformation. Wild-type and transgenic hairy roots contained various phenolic compounds, although both of them had similar concentrations of phenolic compounds. In the future, the protocols described here should be useful for studying and extracting valuable metabolites such as phenolic compounds from P. grandiflorum hairy root cultures.


Subject(s)
Platycodon/genetics , Rhizobium/genetics , Tissue Culture Techniques/methods , Transformation, Genetic/genetics , Chromatography, High Pressure Liquid , DNA Primers/genetics , Kanamycin , Plant Roots/growth & development , Plant Roots/metabolism , Polymerase Chain Reaction , Saponins/isolation & purification , Triterpenes/isolation & purification
17.
Bioresour Technol ; 101(24): 9715-22, 2010 Dec.
Article in English | MEDLINE | ID: mdl-20708926

ABSTRACT

The involvement of genes in flavones biosynthesis was investigated in different organs and suspension cells obtained from Scutellaria baicalensis. Three full-length cDNAs encoding phenylalanine ammonia-lyase isoforms (SbPAL1, SbPAL2, and SbAPL3) and one gene encoding cinnamate 4-hydroxylase (SbC4H) from S. baicalensis were isolated using rapid amplification of cDNA ends (RACE)-PCR. These cDNAs were used together with previously-isolated clones for 4-coumaroyl CoA ligase (4CL) and chalcone synthase (CHS) to show the expression level in different organs of S. baicalensis. These genes were upregulated in suspension cells of S. baicalensis with biotic/abiotic stress factors. The baicalin and baicalein contents in roots were 22 and 107 times higher than those in flowers, respectively. The treatment of suspension cells with methyl jasmonate (MeJa) enhanced the major flavones in S. baicalensis. Cumulatively, the results of this study should advance ability to biosynthesize important and useful medicinal compounds from a variety of plant species.


Subject(s)
Flavones/biosynthesis , Genes, Plant/genetics , Phenylalanine Ammonia-Lyase/genetics , Scutellaria baicalensis/enzymology , Scutellaria baicalensis/genetics , Trans-Cinnamate 4-Monooxygenase/genetics , Amino Acid Sequence , Chromatography, High Pressure Liquid , Cloning, Molecular , Flavanones/biosynthesis , Flavonoids/biosynthesis , Gene Expression Regulation, Plant , Molecular Sequence Data , Organ Specificity/genetics , Phenylalanine Ammonia-Lyase/chemistry , Phenylalanine Ammonia-Lyase/metabolism , Sequence Alignment , Sequence Analysis, DNA , Time Factors , Trans-Cinnamate 4-Monooxygenase/chemistry , Trans-Cinnamate 4-Monooxygenase/metabolism
18.
J Chem Ecol ; 36(8): 914-22, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20614166

ABSTRACT

Sorgoleone, found in the root exudates of sorghum [(Sorghum bicolor (L.) Moench], has been a subject of continued research. Sorgoleone production in grain sorghum roots was investigated under different growth conditions. Methanol was the most effective solvent for extracting sorgoleone from grain sorghum roots. Sorgoleone production is high in young developing plants. The maximum concentration (microg mg(-1) root dry weight) was produced in 5-d-old seedlings; beyond this age, production declined. However, considering both root weight and sorgoleone content per seedling, 10-d-old seedlings had the highest total amounts (microg). Compared with the control, sorgoleone content increased 6.1, 8.6, and 14.2 times when sorghum seeds were treated with auxins, Hoagland solution, and a combination of auxins and Hoagland solution, respectively. Among the innate immunity response elicitors, cellulose (an elicitor of plant origin) stimulated higher sorgoleone production than the others, and it produced 6.2 times more sorgoleone than the control. Combined treatment of sorghum seeds with half strength Hoagland solution and 5 microg ml(-1) of IBA significantly increased both root growth and sorgoleone content in sorghum seedlings.


Subject(s)
Benzoquinones/metabolism , Edible Grain/metabolism , Plant Roots/metabolism , Sorghum/metabolism , Benzoquinones/isolation & purification , Edible Grain/drug effects , Edible Grain/growth & development , Indoleacetic Acids/pharmacology , Lipids/biosynthesis , Lipids/isolation & purification , Plant Roots/drug effects , Plant Roots/growth & development , Seedlings/growth & development , Seedlings/metabolism , Sorghum/drug effects , Sorghum/growth & development , Time Factors
19.
Nat Prod Commun ; 4(7): 903-6, 2009 Jul.
Article in English | MEDLINE | ID: mdl-19731589

ABSTRACT

The ginsenoside content of berries and roots of three cultivars of Korean ginseng have been investigated. For all cultivars, ginsenoside Re was the most abundant ginsenoside in roots and berries. However, berries produced more total ginsenosides, and berry the ginsenoside profile differed from that of roots. The ginsenoside Re content of berries was 4-6 times more than that of roots. Averaged across all cultivars, the amounts of the five ginsenosides in berries was Re > Rc approximately = Rg1 approximately = Rb1 approximately = Rd. For roots, the amounts were Re > Rg1 > Rb1 > Rc > Rd. Roots of the Yunpoong cultivar had the greatest ginsenoside content, followed by roots of the Chunpoong cultivar and the Gumpoong cultivar. The total amount of ginsenosides (especially Rb1, Re, and Rg1) was greatest in the Yunpoong cultivar.


Subject(s)
Ginsenosides/chemistry , Panax/chemistry , Calibration , Chromatography, High Pressure Liquid , Fruit/chemistry , Ginsenosides/isolation & purification , Korea , Plant Roots/chemistry , Reproducibility of Results
20.
J Agric Food Chem ; 57(16): 7513-22, 2009 Aug 26.
Article in English | MEDLINE | ID: mdl-19655726

ABSTRACT

Metabolite profiling and fingerprint analysis by (1)H NMR spectroscopy were used to identify potential biomarkers capable of distinguishing different ginseng species, varieties, and commercial products with the aim of establishing quality control code protocol based on biochemical phenotype. Principal component (PC) analyses of (1)H NMR spectra reliably discriminated between the various ginseng samples, demonstrating the potential utility of metabolomics in the natural health products industry. Four Asian ginseng varieties separated along the PC1 and PC2 axes, and four different Korean ginseng products were divided into two groups by PC1. A strong separation was also revealed between Asian ginseng (Panax ginseng) and American ginseng (Panax quinquefolius). Glutamine, arginine, sucrose, malate, and myo-inositol were the major metabolites in ginseng samples tested in this study. Combined metabolite fingerprinting and profiling suggested that several compounds including glucose, fumarate, and various amino acids could serve as biomarkers for quality assurance in ginseng.


Subject(s)
Drugs, Chinese Herbal/chemistry , Magnetic Resonance Spectroscopy/methods , Metabolomics/methods , Metabolomics/organization & administration , Panax/chemistry , Drugs, Chinese Herbal/metabolism , Panax/metabolism , Quality Control
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