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ABSTRACT: To develop novel and crude anti-obesity drugs from natural products is a promising field to approach the solution to a global health problem such as obesity. The aim of this study was to screen crude anti-obesity drugs from 400 natural products on lipase inhibition activity in vitro.
Among the natural products examined, 31 extracts showed significantly inhibition activity against porcine pancreatic lipase (triacylglycerol lipase, EC 3.1.1.3) by using spectrophotometry with 2,4-dinitrophenylbutyrate as a substrate. Furthermore, 31 natural products were investigated with regard to their lipid inhibition in 3T3-L1 cells. Among these, one of most promising was Nepeta japonica Maximowicz extract, which showed inhibition of triglyceride accumulation in 3T3-L1 adipocytes, suggesting anti-obesity activity. Also, the amount of glycerol released from cells into the medium was increased by treatment of Nepeta japonica Maximowicz extract at a concentration of 100 µg mL(-1) .
The present study suggests that a promising crude anti-obesity drug screened from 400 natural products might be of therapeutic interest with respect to the treatment of obesity.
Journal of the Science of Food and Agriculture 02/2012; 92(10):2195-9. · 1.44 Impact Factor
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ABSTRACT: Obesity is a global health problem. It is also known to be a risk factor for the development of metabolic disorders, type 2 diabetes, systemic hypertension, cardiovascular disease, dyslipidemia, and atherosclerosis. In this study, we screened crude extracts from 400 plants to test their anti-obesity activity using porcine pancreatic lipase assay (PPL; triacylglycerol lipase, EC 3.1.1.3) in vitro activity. Among the 400 plants species examined, 44 extracts from plants, showed high anti-lipase activity using 2,4-dinitrophenylbutyrate as a substrate in porcine pancreatic lipase assay. Furthermore, 44 plant extracts were investigated for their inhibition of lipid accumulation in 3T3-L1 cells. Among these 44 extracts examined, crude extracts from 4 natural plant species were active. Salicis Radicis Cortex had the highest fat inhibitory activity, whereas Rubi Fructus, Corni Fructus, and Geranium nepalense exhibited fat inhibitory capacity higher than 30% at 100 μg/mL in 3T3-L1 adipocytes, suggesting anti-obesity activity. These results suggest that four potent plant extracts might be of therapeutic interest with respect to the treatment of obesity.
International Journal of Molecular Sciences 01/2012; 13(2):1710-9. · 2.60 Impact Factor
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ABSTRACT: Obesity is a global health problem. It is also known to be a risk factor for the development of metabolic disorders, type 2 diabetes, systemic hypertension, cardiovascular disease, dyslipidemia, and atherosclerosis. In this study, we elucidated that Buddleja officinalis Maximowicz extract significantly inhibited lipid accumulation during 3T3-L1 adipocyte differentiation. Furthermore, Buddleja officinalis Maximowicz extract reduced the body weight gain induced through feeding a high-fat diet to C57BL/6 mice. The treatment of Buddleja officinalis Maximowicz extract significantly reduced the adipose tissue weight to 2.7/100 g of body weight in high-fat mice. When their adipose tissue morphology was investigated for histochemical staining, the distribution of cell size in the high-fat diet groups was hypertrophied compared with those from Buddleja officinalis Maximowicz extract-treated mice. In addition, in Buddleja officinalis Maximowicz extract-treated mice, a significant reduction of serum triglyceride and T-cholesterol was observed at to 21% and 17%, respectively. The discovery of bioactive compounds from diet or dietary supplementation is one of possible ways to control obesity and to prevent or reduce the risks of various obesity-related diseases. These results support that Buddleja officinalis Maximowicz extract is expected to create the therapeutic interest with respect to the treatment of obesity.
Molecules 01/2012; 17(7):8687-95. · 2.39 Impact Factor
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ABSTRACT: Ionizing radiation has become an inevitable health concern emanating from natural sources like space travel and from artificial sources like medical therapies. In general, exposure to ionizing radiation such as γ-rays is one of the methods currently used to stress specific model systems. In this study, we elucidated the long-term effect of acute and fractionated irradiation on DCX-positive cells in hippocampal neurogenesis. Groups of two-month-old C57BL/6 female mice were exposed to whole-body irradiation at acute dose (5 Gy) or fractional doses (1 Gy × 5 times and 0.5 Gy × 10 times). Six months after exposure to γ-irradiation, the hippocampus was analyzed. Doublecortin (DCX) immunohistochemistry was used to measure changes of neurogenesis in the subgranular zone (SGZ) of the hippocampal dentate gyrus (DG). The number of DCX-positive cells was significantly decreased in all acute and fractionally irradiation groups. The long-term changes in DCX-positive cells triggered by radiation exposure showed a very different pattern to the short-term changes which tended to return to the control level in previous studies. Furthermore, the number of DCX-positive cells was relatively lower in the acute irradiation group than the fractional irradiation groups (approximately 3.6-fold), suggesting the biological change on hippocampal neurogenesis was more susceptible to being damaged by acute than fractional irradiation. These results suggest that the exposure to γ-irradiation as a long-term effect can trigger biological responses resulting in the inhibition of hippocampal neurogenesis.
Molecules 01/2012; 17(8):9462-8. · 2.39 Impact Factor
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ABSTRACT: Globally, one in three of the World's adults are overweight and one in 10 is obese. By 2015, World Health Organization (WHO) estimates the number of chubby adults will balloon to 2.3 billion--Equal to the combined populations of China, Europe and the United States. The discovery of bioactive compounds from herbs is one possible way to control obesity and to prevent or reduce the risks of developing various obesity-related diseases. In this study, we screened anti-obesity agents such as methyl gallate from the herbal composition known as HemoHIM that actively inhibits lipid formation as evidenced by Oil Red O staining and triglyceride (TG) contents in 3T3-L1 adipocytes, suggesting their use as an anti-obesity agent. Furthermore, the amount of glycerol released from cells into the medium had increased by treatment of methyl gallate in a concentration-dependent manner. The present study suggests that a promising anti-obesity agent like methyl gallate might be of therapeutic interest for the treatment of obesity.
Molecules 01/2012; 17(4):3630-8. · 2.39 Impact Factor
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ABSTRACT: In this study, we elucidated a small molecule inhibitor on viral protein NS5B identified through a high-throughput screening strategy using optical nanoparticle-based RNA oligonucleotide. We have previously shown that quantum dots (QDs)-RNA oligonucleotide can specifically recognize the HCV viral proteins. We have also demonstrated that conjugated QDs-RNA oligonucleotide can specifically and sensitively interact with designed biochips [1,2]. Among the flavonoids examined, (-)-epigallocatechin gallate (EGCG) demonstrated a remarkable inhibition activity on HCV viral protein, NS5B. (-)-Epigallocatechin gallate, at 0.005 μg mL(-1) or more, concentration-dependently attenuated the binding affinity on a designed biochip as evidenced by QDs-RNA oligonucleotide. At a concentration of 0.1 μg mL(-1), (-)-epigallocatechin gallate showed a 50% inhibition activity on QDs-RNA oligonucleotide biochip assay. We screened a small molecule inhibitor on the viral protein, NS5B, identified through a high-throughput screening strategy using on-chip optical nanoparticle-based RNA oligonucleotide on chip. In this designed strategy, the convenient and efficient screening and development of an on-chip viral protein inhibitor using a QDs-RNA oligonucleotide assay is achievable with high sensitivity and simplicity. In addition, this platform is expected to be applicable toward the inhibitor screening of other types of diseases.
Talanta 10/2011; 85(5):2639-42. · 3.79 Impact Factor
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ABSTRACT: To investigate changes in gonadal white adipose tissue and lipogenesis-related gene expression induced by radiation exposure.
Groups of two-month-old C57BL/6 mice were exposed whole-body to ¹³⁷Cs γ-rays at a single dose (5 gray [Gy]) or fractionated doses (1 Gy x 5 times, 0.5 Gy x 10 times, or 0.2 Gy x 25 times). Six months after irradiation, gonadal white adipose tissue was isolated from mice. Two and 25-month-old mice were used as young and old study references. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was used to measure messenger RNA (mRNA) expression of genes related to: (i) Primary lipid metabolism (ATP-citrate lyase [ACL], malic enzyme1 [ME1] and glucose-6-phosphate dehydrogenase 2 [G6PD2]), (ii) glucose uptake (glucose transporter 4 [GLUT4]), (iii) fatty acid synthesis (sterol regulatory element binding transcription factor 1 [SREBP-1c], fatty acid synthetase [FAS] and acetyl-coenzyme A carboxylase beta [ACC]), (iv) triglyceride synthesis (diacylglycerol O-acyltransferase 1 [DGAT1] and diacylglycerol O-acyltransferase 2 [DGAT2]), and (v) adipose-derived hormones (leptin [LEP]).
The weight of gonadal white adipose tissue in the irradiated groups tended to increase compared to the non-irradiated group though the radiation-induced increase in white adipose tissue was only significant for the 5 x 1 Gy group. The mRNA levels of SREBP-1c, ACC, FAS, ACL, GLUT4, ME1 and G6PD2 were relatively lower in γ-irradiated groups than in non-irradiated groups. The mRNA levels of leptin and DGAT were relatively higher than non-irradiated groups. The changes in expression of these lipogenesis-related genes caused by γ-irradiation showed a very similar pattern to changes caused by ageing.
A physical agent such as γ-rays can trigger biological responses resulting in fat accumulation of gonadal white adipose tissue in mice.
International Journal of Radiation Biology 01/2011; 87(3):302-10. · 2.28 Impact Factor
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ABSTRACT: In this study, we elucidated the inhibitory effect of fucoidan from marine brown algae on the lipid accumulation in differentiated 3T3-L1 adipocytes and its mechanism. The treatment of fucoidan in a dose-dependent manner was examined on lipid inhibition in 3T3-L1 cells by using Oil Red O staining. Fucoidan showed high lipid inhibition activity at 200 μg/mL concentration (P < 0.001). Lipolytic activity in adipocytes is highly dependent on hormone sensitive lipase (HSL), which is one of the most important targets of lipolytic regulation. Here, we examined the biological response of fucoidan on the protein level of lipolysis pathway. The expressed protein levels of total hormone sensitive lipase (HSL) and its activated form, phosphorylated-HSL were significantly increased at concentration of 200 μg/mL fucoidan. Furthermore, insulin-induced 2-deoxy-D-[³H] glucose uptake was decreased up to 51% in fucoidan-treated cells as compared to control. Since increase of HSL and p-HSL expression and decrease of glucose uptake into adipocytes are known to lead to stimulation of lipolysis, our results suggest that fucoidan reduces lipid accumulation by stimulating lipolysis. Therefore, these results suggest that fucoidan can be useful for the prevention or treatment of obesity due to its stimulatory lipolysis.
Marine Drugs 01/2011; 9(8):1359-67. · 3.85 Impact Factor
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ABSTRACT: Globally, over 170 million people (ca. 3% of the World's population) are infected with the hepatitis C virus (HCV), which can cause serious liver diseases such as chronic hepatitis, evolving into subsequent health problems. Driven by the need to detect the presence of HCV, as an essential factor in diagnostic medicine, the monitoring of viral protein has been of great interest in developing simple and reliable HCV detection methods. Despite considerable advances in viral protein detection as an HCV disease marker, the current enzyme linked immunosorbent assay (ELISA) based detection methods using antibody treatment have several drawbacks. To overcome this bottleneck, an RNA aptamer become to be emerged as an antibody substitute in the application of biosensor for detection of viral protein. In this study, we demonstrated a streptavidin-biotin conjugation method, namely, the RNA aptamer sensor system that can quantify viral protein with detection level of 700 pg mL(-1) using a biotinylated RNA oligonucleotide on an Octet optical biosensor. Also, we showed this method can be used to screen inhibitors of viral protein rapidly and simply on a biotinylated RNA oligonucleotide biosensor. Among the inhibitors screened, (-)-Epigallocatechin gallate showed high binding inhibition effect on HCV NS5B viral protein. The proposed method can be considered a real-time monitoring method for inhibitor screening of HCV viral protein and is expected to be applicable to other types of diseases.
Sensors 01/2011; 11(7):6685-96. · 1.74 Impact Factor
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ABSTRACT: Globally, there are concerns over the risks associated with radiation exposure, so it is important to understand the biological effects of radiation exposure. Driven by the need to detect the presence of radiation exposure, biomarkers to monitor potential exposure after radiological accidents can be developed and would be extremely valuable for biological response. In this study, the behavior of spermidine as a biomarker was investigated in a C57BL/6 mouse model exposed to an acute whole-body sublethal dose of 6 Gy. The spermidine content values in serum increased for up to two days after 6 Gy irradiation. However, the enhanced spermidine content observed on day +3 in irradiated mice returned to normal levels on the subsequent five days. The result indicates that spermidine can be used as a biomarker of biological response to radiation exposure.
Molecules 01/2011; 17(1):145-50. · 2.39 Impact Factor
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ABSTRACT: In this study, we showed that γ-irradiation could trigger biological response like the fat accumulation of gonadal white adipose tissue (gWAT) in mice. The irradiated animal can be used as a useful model for evaluation of anti-obesity drugs. To induce the fat accumulation by γ-irradiation, 2-months-old female C57BL/6 mice were irradiated at 5 Gy and further raised for 6 months. Subsequently, the mice were i.p. injected daily with Orlistat (25 mg/kg) or vehicle for 3 wk and analyzed for the adipose tissue weight and serum triglyceride and T-cholesterol levels. The abdominal WAT of the γ-irradiated mice weighed an average of 3.9/100 g body weight, 1.7-fold higher than what was seen in normal mice (2.3/100 g body weight), indicating that γ-irradiation induced the fat accumulation in the adipose tissue. However, the administration of Orlistat, a well-known anti-obesity agent, significantly reduced the adipose tissue weight to 1.7/100 g body weight in irradiated mice. In addition, in these Orlistat-treated mice, a significant reduction of serum triglyceride and T-cholesterol was observed up to 14% and 27%, respectively.
European Journal of Lipid Science and Technology 12/2010; 112(12):1384 - 1388. · 1.73 Impact Factor
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ABSTRACT: BACKGROUND: Over 170 million people, more than 3% of the world's population, suffer from the hepatitis C virus (HCV) infection and the rate of death from liver-related mortality to HCV has increased. In respect of this, the development of assays for biological imaging should be urgently considered as an essential factor in diagnosis.RESULTS: A novel HCV-detecting technique using a nanoparticle-supported aptamer probe was demonstrated. With the aid of nanoparticle quantum dots (QDs) with carboxyl group as an imaging probe, and 5′-end-amine-modified RNA oligonucleotide as a capturing probe, target HCV NS3 was visually detected on chip. The QDs-based RNA aptamer for HCV NS3 showed high selectivity and specificity against other protein such as BSA. The detection limit of HCV NS3 protein was 5 ng mL−1 level.CONCLUSION: With a novel strategy for protein–aptamer interaction, the feasibility of applying QDs-based fluorescent detection technique to HCV viral protein assay for the development of a protein biochip was demonstrated. This scheme of QDs-mediated imaging with a target-oriented specific RNA aptamer for the detection of infectious HCV diseases provides an efficient strategy and a promising new platform for monitoring applications. Copyright © 2010 Society of Chemical Industry
Journal of Chemical Technology & Biotechnology 07/2010; 85(8):1130 - 1134. · 2.17 Impact Factor
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ABSTRACT: Globally, approximately 170 million people (representing approximately 3% of the population worldwide), are infected with hepatitis C virus (HCV) and at risk of serious liver disease, including chronic hepatitis. We propose a new quantum dots (QDs)-supported RNA oligonucleotide approach for the specific and sensitive detection of viral protein using a biochip. This method was developed by immobilizing a HCV nonstructural protein 5B (NS5B) on the surface of a glass chip via the formation of a covalent bond between an amine protein group and a ProLinker glass chip. The QDs-supported RNA oligonucleotide was conjugated via an amide formation reaction from coupling of a 5'-end-amine-modified RNA oligonucleotide on the surface of QDs displaying carboxyl groups via standard EDC coupling. The QDs-conjugated RNA oligonucleotide was interacted to immobilized viral protein NS5B on the biochip. The detection is based on the variation of signal of QDs-supported RNA oligonucleotide bound on an immobilized biochip. It was demonstrated that the value of the signal has a linear relationship with concentrations of the HCV NS5B viral protein in the 1 microg mL(-1) to 1 ng mL(-1) range with a detection limit of 1 ng mL(-1). The major advantages of this RNA-oligonucleotide nanoparticle assay are its good specificity, ease of performance, and ability to perform one-spot monitoring. The proposed method could be used as a general method of HCV detection and is expected to be applicable to other types of diseases as well.
International Journal of Nanomedicine 01/2010; 5:323-9. · 3.13 Impact Factor
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ABSTRACT: Regiospecific 3'-hydroxylation reaction of daidzein was performed with CYP105D7 from Streptomyces avermitilis MA4680 expressed in Escherichia coli. The apparent K(m) and k(cat) values of CYP105D7 for daidzein were 21.83 +/- 6.3 microM and 15.01 +/- 0.6 min(-1) in the presence of 1 microM of CYP105D7, putidaredoxin (CamB) and putidaredoxin reductase (CamA), respectively. When CYP105D7 was expressed in S. avermitilis MA4680, its cytochrome P450 activity was confirmed by the CO-difference spectra at 450 nm using the whole cell extract. When the whole-cell reaction for the 3'-hydroxylation reaction of daidzein was carried out with 100 microM of daidzein in 100 mM of phosphate buffer (pH 7.5), the recombinant S. avermitilis grown in R2YE media overexpressing CYP105D7 and ferredoxin FdxH (SAV7470) showed a 3.6-fold higher conversion yield (24%) than the corresponding wild type cell (6.7%). In a 7 L (working volume 3 L) jar fermentor, the recombinants S. avermitilis grown in R2YE media produced 112.5 mg of 7,3',4'-trihydroxyisoflavone (i.e., 29.5% conversion yield) from 381 mg of daidzein in 15 h.
Biotechnology and Bioengineering 10/2009; 105(4):697-704. · 3.95 Impact Factor
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ABSTRACT: Screening of bacterial whole cells was performed for regioselective hydroxylation of daidzein and genistein. Among the strains examined, Streptomyces avermitilis MA-4680 showed high ortho-dihydroxylation activity to produce 3',4',7-trihydroxyisoflavone and 3',4',5,7-tetrahydroxyisoflavone from daidzein (4',7-dihydroxyisoflavone) and genistein (4',5,7-trihydroxyisoflavone), respectively. Using 100 mg cells (wet wt.) and 1% (v/v) Triton X100 in 1 ml of total reaction volume, where 100 microl of the substrate solution (0.5 mM in 10% (v/v) mixed solvent of DMSO:MeOH = 3:7) was added to 900 microl of potassium phosphate buffer (100 mM, pH 7.2), a 16% molar conversion yield of 3',4',7-trihydroxyisoflavone was obtained from 0.5 mM daidzein after 24 h of reaction time at 28 degrees C and 200 rpm. Ketoconazole significantly (ca. 90%) inhibited the ortho-hydroxylation activity of daidzein, suggesting that cytochrome P450 enzymes putatively play roles in regiospecific daidzein hydroxylation. The analysis of the reaction products was determined by gas chromatography/mass spectrometry (GC/MS) and (1)H NMR
Journal of Molecular Catalysis B Enzymatic 09/2009; 59(4). · 2.73 Impact Factor
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ABSTRACT: Screening of bacterial whole cells was performed for regioselective hydroxylation of daidzein and genistein. Among the strains examined, Streptomyces avermitilis MA-4680 showed high ortho-dihydroxylation activity to produce 3',4',7-trihydroxyisoflavone and 3',4',5,7-tetrahydroxyisoflavone from daidzein (4',7-dihydroxyisoflavone) and genistein (4',5,7-trihydroxyisoflavone), respectively. Using 100 mg cells (wet wt.) and 1% (v/v) Triton X100 in 1 ml of total reaction volume, where 100 microl of the substrate solution (0.5 mM in 10% (v/v) mixed solvent of DMSO:MeOH = 3:7) was added to 900 microl of potassium phosphate buffer (100 mM, pH 7.2), a 16% molar conversion yield of 3',4',7-trihydroxyisoflavone was obtained from 0.5 mM daidzein after 24 h of reaction time at 28 degrees C and 200 rpm. Ketoconazole significantly (ca. 90%) inhibited the ortho-hydroxylation activity of daidzein, suggesting that cytochrome P450 enzymes putatively play roles in regiospecific daidzein hydroxylation. The analysis of the reaction products was determined by gas chromatography/mass spectrometry (GC/MS) and (1)H NMR.
Journal of Bioscience and Bioengineering 08/2009; 108(1):41-6. · 1.79 Impact Factor
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ABSTRACT: Metagenomic DNA (gDNA) is an important research topic because uncultivated microorganisms represent an interesting reservoir of genes with potential biotechnological applications. Here we describe a novel Si-C-N chromatography approach of gDNA extraction from environmental samples. Amorphous Si-C-N ceramic was obtained by the pyrolysis of a polymer precursor (polycarbosilazane) at 1350 degrees C in Ar. The purified gDNA fragments were at least 12 kilo base pairs in size and were sufficiently free of contaminants, thus were applicable to both restriction enzyme digestion using five different enzymes and polymerase chain reaction amplification for detecting phylogenetic groups of native microorganisms in environmental samples. This Si-C-N material produces pure gDNA that can be utilized in some of the most common molecular biological procedures applied in the purification step.
Journal of Chromatography 03/2008; 1179(2):145-51. · 4.53 Impact Factor
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ABSTRACT: To analyze a cotG-based Bacillus subtilis spore display system directly, GFP(uv) was expressed on the surface of Bacillus subtilis spores. When GFP(uv) was fused to the C-terminal of the cotG structural gene and expressed, the existence of a CotG-GFP(uv) fusion protein on the B. subtilis spore was confirmed by flow cytometry confocal microscopic analysis. When the cotG anchoring motif was deleted, no fluorescence emission was observed under flow cytometry and confocal microscopic analysis from the purified spore, confirming the essential role of CotG as an anchoring motif. This GFP(uv) displaying spore might be used for another signaling application triggered by intracellular or extracellular stimuli.
Journal of Microbiology and Biotechnology 05/2007; 17(4):677-80. · 1.38 Impact Factor
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ABSTRACT: An important prerequisite for successful construction of a metagenome library is an efficient procedure for extracting DNA from environmental samples. We compared three indirect and four direct extraction methods, including a commercial kit, in terms of DNA yield, purity, and time requirement. A special focus was on methods that are appropriate for the extraction of environmental DNA (eDNA) from very limited sample sizes (0.1 g) to enable a highly parallel approach. Direct extraction procedures yielded on average 100-fold higher DNA amounts than indirect ones. A drawback of direct extraction was the small fragment size of approx 12 kb. The quality of the extracted DNA was evaluated by the ability of different restriction enzymes to digest the eDNA. Only the commercial kit and a direct extraction method using freeze-thaw cell lysis in combination with an in-gel patch electrophoresis with hydroxyapatite to remove humic acid substances yielded DNA, which was completely digested by all restriction enzymes. Moreover, only DNA extracted by these two procedures could be used as template for the amplification of fragments of several 16S rDNA, 18S rDNA groups under standard polymerase chain reaction conditions.
Applied Biochemistry and Biotechnology 09/2006; 134(2):97-112. · 1.94 Impact Factor
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ABSTRACT: Most of the microorganism species are largely untapped and could represent an interesting reservoir of genes useful for biotechnological applications. Unfortunately, a major difficulty associated with the methods used to isolate environmental DNA is related to the contamination of the extracted material with humic substances. These polyphenolic compounds inhibit the DNA processing reactions and severely impede cloning procedures. In this work, we describe a rapid, simple, and efficient method for the purification of genomic DNA from environmental samples: we added a chromatography step directly embedded into an agarose gel electrophoresis. This strategy enabled the DNA extraction from various environmental samples and it appeared that the purity grade was compatible with digestion by restriction enzymes and polymerase chain reaction (PCR) amplifications.
Electrophoresis 09/2005; 26(16):3055-61. · 3.30 Impact Factor
