• ALGAE
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• 제27권2호
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• pp.125-134
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• 2012

Germanium dioxide ($GeO_2$) has been used for many years in the cultivation of red and green algae as a means of controlling the growth of diatoms. Brown algae are sensitive to $GeO_2$, however, the basis of this sensitivity has not been characterized. Here we use embryos of $Fucus$ $vesiculosus$ to investigate morphological and physiological impacts of $GeO_2$ toxicity. Morphometric features of embryos were measured microscopically, and physiological features were determined using pulse amplitude modulated (PAM) fluorometry. At 5 mg $L^{-1}$ $GeO_2$, embryos grew slower than controls and developed growth abnormalities. After 24 h, initial zygote divisions were often oblique rather than transverse. Rhizoids had inflated tips in $GeO_2$ and were less branched, and apical hairs were deformed, with irregularly aligned, spheroidal cells. Minimum fluorescence ($F_0$) showed minor differences over the 10 days experiment, and pigment levels (chlorophylls $a$, $c$ and total carotenoids) showed no difference after 10 days. Optimum quantum yield increased from ca. 0.52 at 24 h to 0.67 at 5 days, and $GeO_2$-treated embryos had higher mean values (significant at 3 and 5 days). Optimum quantum yield of photosystem II (${\Phi}_{PSII}$) was stable in control thalli after 5 days, but declined significantly in $GeO_2$. Addition of silica (as $SiO_2$) did not reverse the effects of $GeO_2$. These results suggest that $GeO_2$ toxicity in brown algae is associated with negative impacts at the cytological level rather than metabolic impacts associated with photosynthesis.

• Nutrition Research and Practice
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• 제8권5호
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• pp.550-557
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• 2014

BACKGROUND/OBJECTIVES: Angelica keiskei is a green leafy vegetable rich in plant pigment phytochemicals such as flavonoids and carotenoids. This study examined bioavailability of flavonoids and carotenoids in Angelica keiskei and the alteration of the antioxidant performance in vivo. SUBJECTS AND MATERIALS: Absorption kinetics of phytochemicals in Angelica keiskei were determined in healthy older adults (> 60 y, n = 5) and subjects with metabolic syndrome (n = 5). Subjects consumed 5 g dry Angelica keiskei powder encapsulated in gelatin capsules with a low flavonoid and carotenoid liquid meal. Plasma samples were collected at baseline, 0.5, 1, 2, 3, 4, 5, 6, 7, and 8 h. Samples were analyzed for flavonoids and carotenoids using HPLC systems with electrochemical and UV detection, respectively, and for total antioxidant performance by fluorometry. RESULTS: After ingestion of Angelica keiskei increases in plasma quercetin concentrations were observed at 1-3 and 6-8 hr in the healthy group and at all time points in the metabolic syndrome group compared to baseline (P < 0.05). Plasma lutein concentrations were significantly elevated in both the healthy and metabolic syndrome groups at 8 hr (P < 0.05). Significant increases in total antioxidant performance were also observed in both the healthy and the metabolic syndrome groups compared to baseline (P < 0.05). CONCLUSIONS: Findings of this study clearly demonstrate the bioavailability of phytonutrients of Angelica keiskei and their ability to increase antioxidant status in humans.

• BMB Reports
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• 제38권1호
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• pp.104-110
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• 2005

We extended the measurable time scale of DNA dynamics to microsecond using $[Ru(phen)_2(dppz)]^{2+}$ (phen = 1,10-phenanthroline, dppz = dipyrido[3,2-a:2',3'-c]phenazine) (RuPD), which displays a mean lifetime near 500 ns. To evaluate the usefulness of this luminophore (RuPD) for probing nucleic acid dynamics, its intensity and anisotropy decays when intercalated into supercoiled and linear pBluescript (pBS) II SK(+) phagemids were examined using frequency-domain fluorometry with a blue light-emitting diode (LED) as the modulated light source. The mean lifetime for the supercoiled phagemids (< $\tau$ > = 489.7 ns) was somewhat shorter than that for the linear phagemids (< $\tau$ > = 506.4 ns), suggesting a more efficient shielding from water by the linear phagemids. The anisotropy decay data also showed somewhat shorter slow rotational correlation times for supercoiled phagemids (997.2 ns) than for the linear phagemids (1175.6 ns). The slow and fast rotational correlation times appear to be consistent with the bending and torsional motions of the phagemids, respectively. These results indicate that RuPD can have applications in studies of both bending and torsional dynamics of nucleic acids.

• Reproductive and Developmental Biology
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• 제28권3호
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• pp.191-196
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• 2004

본 실험에서는 외래 유전자의 효율적인 발현을 위하여 GFP 표지유전자를 이용하여 여러 종류의 promoter를 검정하였다. 또한, retrovirus vector에 WPRE 서열을 도입함으로써 GFP 유전자의 발현 증가 여부를 확인하였다. 모든 표적 세포에 있어서 UbC와 β-actin promoter에 비해 RSV와 CMV promoter 통제하의 GFP의 발현이 더 강하게 나타났으며, 특히 CEF 세포에서는 RSV promoter가 가장 우수한 것으로 확인되었다. WPRE의 도입으로 인한 발현율의 증가는 CEF를 제외한 세포주에서 promoter의 종류에 관계없이 확인되었다. 이상의 결과로 각 세포주는 promoter에 따라 발현 양상이 약간의 차이를 보이고 있으나 RSV와 CMV promoter에서 유전자의 발현이 보다 효율적이며, WPRE 서열이 도입된 경우에 HeLa와 PFF 세포에서 발현이 현저히 증가하는 것을 확인할 수 있었다. 이러한 연구 결과는 효율적인 유전자의 발현 체계를 확립하는데 기여함으로써 더 나아가 유전자 치료나 형질전환 동물생산에 적극적으로 활용되어질 수 있을 것이다.

• BMB Reports
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• 제35권4호
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• pp.389-394
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• 2002

$[Ru(bpy)_2(dppz)]^2+$ (bpy=2,2'-bipyfidine, dppz=dipyrido[3,2-a:2',3'-c]phenazine) (RuBD), a long-lifetime metal-ligand complex, displays favorable photophysical properties. These include long lifetime, polarized emission, but no significant fluorescence from the complex that is not bound to DNA. To show the usefulness of this luminophore (RuBD) for probing the bending and torsional dynamics of nucleic acids, its intensity and anisotropy decays when intercalated into supercoiled and relaxed pTZ18U plasmids were examined using frequency-domain fluorometry with a blue light-emitting diode (LED) as the modulated light source. The mean lifetimes for the supercoiled plasmids (< $\tau$ >=148 ns) were somewhat shorter than those for the relaxed plasmids (< $\tau$ >=160 ns). This suggests that the relaxed plasmids were shielded more efficiently from water. The anisotropy decay data also showed somewhat shorter slow rotational correlation times for supercoiled plasmids (288 ns) than for the relaxed plasmids (355 ns). The presence of two rotational correlation times suggests that RuBD reveals both the bending and torsional motions of the plasmids. These results indicate that RuBD can be useful for studying both the bending and torsional dynamics of mucleic acids.

• 대한한방내과학회지
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• 제24권1호
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• pp.75-83
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• 2003

Objectives : Peroxynitrite($ONOO^-$), formed from the reaction of $O2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been involved in the aging process and age-related diseases such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors, NO and $O_{2^-}$ and its scavenging mechanism of Zingiberis Rhizoma (ZR). Methods : To investigate scavenging activities of $ONOO^-,\;NO,\;O_{2^-}$ and its scavenging mechanism, we used fluorescent probes like DCFDA, DAF-2 and DHR 123. The $ONOO^-$ scavenging activity on ZR was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorometry. The scavenging efficacy was expressed as IC50, showing the concentration of each sample that is required to cause 50% inhibition of DHR 123 oxidation. In a separate study, the protective effect of ZR on $ONOO^-$-induced nitration of bovine serum albumin was investigated through immuno-assay with a monoclonal anti-nitryrosine antibody, and a horseradish peroxidase-conjugated anti-mouse secondary antibody from sheep. Results : ZR markedly scavenged authentic $ONOO^-,\;O_{2^-}$ and NO. It also inhibited $ONOO^-$ induced by $O_{2^-}$ and NO which are derived from SIN-1. The data demonstrated that ZR led to decreased $ONOO^-$ mediated nitration of tyrosine through electron donation. It also inhibited the nitration of bovine serum albumin induced by $ONOO^-$ in a dose-dependent manner. Furtheremore, it blocked LPS-induced ROS and RNS generation. Conclusions : These results suggest that ZR can be developed as an effective $ONOO^-$ scavenger for the prevention of aging process and age-related diseases.

• 대한한방내과학회지
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• 제26권1호
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• pp.107-118
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• 2005

Objectives : Peroxynitrite $(ONOO^-)$, fonned from the reaction of $O_2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in the aging process and age-related disease such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. Due to the lack of endogenous enzymes to thwart $ONOO^-$ activation, developing a specific $ONOO^-$ scavenger is remarkably important. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors, NO and $O_2^-$ and its scavenging mechanism of Ojawhan. Methods : To investigate scavenging activities of $ONOO^-$, NO, $O_2^-$ and its scavenging mechanism using fluorescent probes, DCFDA, DAF-2 and DHR 123. The $ONOO^-$ scavenging activity on Ojawhan was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorometry. Oxidative stress was induced by strong oxidants t-butyl hydroperoxide (t-BHP). Endothelial cell (YPEN-1) was used for detection of intracellular oxidative stress. Results : Ojawhan markedly scavenged authentic $ONOO^-$, $O_2^-$ and NO. It also inhibited $ONOO^-$ induced by $O_2^-$ and NO which are derived from SIN-1. Furthennore, ${\underline{Ojawhan}}$ blocked lipopolysaccharide (LPS)-induced $ONOO^-$, $O_2^-$ and NO generation utilizing kidney homogenates of LPS-injected mouse and inhibited t-BHP-induced ROS and $ONOO^-$ in endothelial cell culture system. Conclusions : These results suggest that Ojawhan be developed as an effective $ONOO^-$ scavenger for the prevention of $ONOO^-$ involved diseases and age-related diseases.

• Archives of Pharmacal Research
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• 제25권2호
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• pp.143-150
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• 2002

Fluorescent probes bound to DNA typically display nanosecond decay times and reveal only nanosecond motions. We extend the time range of measurable DNA dynamics using $[Ru(pby)_2(dppz)]^{2+}$ (bpy=2.2'-bipyridine, dppz=dipyrido[3,2-a2',3'-c]phenazine) (RuBD) which displays a mean lifetime near 90 ns. To test the usefulness of RuBD as a probe for diffusive processes in calf thymus DNA, we compared the efficiencies of fluorescence resonance energy transfer (FRET) using three donors which display lifetimes near 5 ns for acridine orange (AO), 22 ns for ethidum bromide (EB) and 92 ns for RuBD, with nile blue (NB) as the acceptor. The F rster distances for AO-NB, EB-NB and RuBD-NB donor-acceptor pairs were 42.3, 52.3, and $30.6{\;}{\AA}$, respectively. All three donors showed dramatic decreases in fluorescence intensities and more rapid intensity decays with increasing NB concentrations. The intensity decays of AO and EB in the presence of varying concentrations of NB were satisfactorily described by the one-dimensional FRET model without diffusion (Blumen and Manz, 1979). In the case of the long-lifetime donor RuBD, the experimental phase and modulation somewhat deviated from the recovered values computed from this model. The recovered NB concentrations and FRET efficiencies from the model were slightly larger than the expected values, however, the recovered and expected values did not show a significant difference. Thus, it is suggested that the lifetime of RuBD is too short to measure diffusive processes in calf thymus DNA.

• Journal of Photoscience
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• 제11권3호
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• pp.133-139
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• 2004

The metal-ligand complex, $[Ru(phen)_2(dppz)]^{2+}$ (phen=1,10-phenanthroline, dppz=dipyrido[3,2-a:2',3'-c]phenazine) (RuPD), was used as a spectroscopic probe for studying nucleic acid dynamics. The RuPD complex displays a long lifetime and a molecular light switch property upon DNA binding due to shielding of its dppz ligand from water. To show the usefulness of this luminophore (RuPD) for probing nucleic acid dynamics, we compared its intensity and anisotropy decays when intercalated into the $tRNA^{val}$ and pBluescript (pBS) II SK(+) phagemid through a comparison with ethidium bromide (EB), a conventional nucleic acid probe. We used frequency-domain fluorometry with a blue light-emitting diode (LED) as the modulated light source. The mean lifetime for the $tRNA^{val}$ (<${\tau}$> = 166.5 ns) was much shorter than that for the pBS II SK(+) phagemid (<${\tau}$> = 481.3 ns), suggesting a much more efficient shielding from water by the phagemid. Because of their size difference, the anisotropy decay data showed a much shorter rotational correlation times for the $tRNA^{val}$ (99.9 and 23.6 ns) than for the pBS II SK(+) phagemid (968.7 and 39.5 ns). These results indicate that RuPD can be useful for studying nucleic acid dynamics.

• Journal of Ginseng Research
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• 제40권4호
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• pp.351-358
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• 2016

Background: This study was designed to investigate whether ginsenoside Rb1 (Rb1) and compound K (CK) ameliorated insulin resistance by suppressing endoplasmic reticulum (ER) stress-induced inflammation in adipose tissue. Methods: To induce ER stress, epididymal adipose tissue from mice or differentiated 3T3 adipocytes were exposed to high glucose. The effects of Rb1 and CK on reactive oxygen species production, ER stress, TXNIP/NLRP3 inflammasome activation, inflammation, insulin signaling activation, and glucose uptake were detected by western blot, emzyme-linked immunosorbent assay, or fluorometry. Results: Rb1 and CK suppressed ER stress by dephosphorylation of $IRE1{\alpha}$ and PERK, thereby reducing TXNIP-associated NLRP3 inflammasome activation in adipose tissue. As a result, Rb1 and CK inhibited IL-$1{\beta}$ maturation and downstream inflammatory factor IL-6 secretion. Inflammatory molecules induced insulin resistance by upregulating phosphorylation of insulin receptor substrate-1 at serine residues and impairing insulin PI3K/Akt signaling, leading to decreased glucose uptake by adipocytes. Rb1 and CK reversed these changes by inhibiting ER stress-induced inflammation and ameliorating insulin resistance, thereby improving the insulin IRS-1/PI3K/Akt-signaling pathway in adipose tissue. Conclusion: Rb1 and CK inhibited inflammation and improved insulin signaling in adipose tissue by suppressing ER stress-associated NLRP3 inflammation activation. These findings offered novel insight into the mechanism by which Rb1 and CK ameliorate insulin resistance in adipose tissue.