Major egg white proteins and their hydrolysates serve as functional food ingredients that have certain metal-chelating properties. Employing egg white proteins and their hydrolysates to scavenge dietary oxalates is anticipated to have beneficial effect in the prevention of kidney stones. The objective of this study was to determine the biogenic oxalate-chelating activity of ovalbumin, ovomucin, and ovotransferrin and their hydrolysates. To prepare oxalate extracts, 30 mL of 0.25 N HCl was added to separately to 0.5 g of dried spinach and starfruit powders followed by boiling for 15 min, and after cooling, the addition of a further 20 mL of 0.25 N HCl. Having prepared these extracts, ovalbumin, ovomucin, and ovotransferrin and their hydrolysates were separately mixed with oxalate extracts and incubated at 3℃ for 24 h. Following centrifugation, supernatants were analyzed by HPLC using a reverse-phase C18 column coupled with a diode array detector. We found that all assessed proteins and their hydrolysates showed biogenic oxalate-chelating activity against the oxalates of spinach. In contrast, however, only ovalbumin, ovalbumin-hydrolysate, and ovomucin showed chelating activity (57.10%±8.84%, 85.44%±5.30%, 73.20%±4.13%, respectively) against the oxalates of starfruit (P<0.05). Overall, hydrolyzed ovalbumin was identified as the most effective chelator of the oxalates both spinach and starfruit. In this study, we thus established that the assessed egg white proteins and their hydrolysates have oxalate-chelating activity in vitro, thereby indicating that these compounds have potential utility as nutraceuticals for the chelation of dietary oxalate. However, further research will be necessary to verify their oxalate-chelating activities against different fruits and vegetables and under specific in vivo conditions and against purified oxalate.
Protein modification by N-acetyl-${\beta}$-D-glucosamine (O-G1cNAc) on the hydroxyl groups of Ser or Thr ubiq-uitously occurs in eukaryotic cells and is involved in many cellular phenomena. The level of O-G1cNAc-mod-ified protein is regulated by OGT and O-GlcNAcase enzymes. We have tried to produce recombinant O-GlcNAcase in E. coli as an effort to establish in vitro screening system for modulators of O-GlcNAcase. The culture conditions for improvement of O-GlcNAcase productivity, were as follows: induction temperature, $30^{\circ}C$; the concentration of L-arabinose, 0.02% and induction time, 5 hr. Under these culture conditions, E. coli cells containing O-GlcNAcase gene had no enzyme activity until up to 3 hr culture. However, O-GlcNAcase activity dramatically increased from 3 to 5 hr culture. It almost maintained the same level after 5 hr culture. Western blot analysis verified the amount of expressed O-GlcNAcase increased with culture time, being con-sistent with activity data. The optimal reaction condition determined in this study was as follows: protein quan-tity, $5{\mu}g$; reaction time, 30 min; reaction temperature, $45^{\circ}C$; substrate concentration, 2 mM; reaction pH, 6.5. Methanol had little effect on O-GlcNAcase activity and 90% of activity were retained at 10%. Only 15% resid-ual activity were detected at 5% of chloroform.
This study investigates the toxic effects of $Cd^{2+}$on frog (Rana dybowskii) by the determination of oocyte maturation and development of embryo exposure to different concentrations of the toxicant. The results show that $Cd^{2+}$ concentration of 0.1ppm suppressed the maturation of the oocytes. To examine the reversibility of the inhibitory effects, the oocytes were exposed to the $Cd^{2+}$ only for 3 hours, and then transferred to plain medium and cultured further for 17 hours. The oocytes were recovered from the toxic effect of the $Cd^{2+}$ when they were exposed to 1ppm, but not to 2.5ppm of the $Cd^{2+}$. The development of 2 cell embryos to 32 cell was completely suppressed at 0.1ppm and the longer the embryos were exposed to the $Cd^{2+}$, the more damage appeared to the embryos and the cytolysis of the 32 cell was induced by $Cd^{2+}$ at 0.1ppm. On the other hand, the embryos of blastula stage were cultured 96 hours in presence of the $Cd^{2+}$ at various concentrations and were examined. The rates of mortality and malformed larvae were investigated by probit analysis. From the results of LC$_{50}$ of 0.1ppm and EC$_{50}$ of 0.08ppm, Tl of 5.0 was derived, which indicates $Cd^{2+}$ is to be considered a teratogenic compound. Such specific malformations occurred in 14.3% as spine deformations at the 0.05ppm, in 75.0% as tail deformations at the 0.1ppm, in 66.7% as abdominal deformations at the 0.01ppm and in 26.0% as profound deformations at the 0.1ppm of $Cd^{2+}$ concentration which living embryos were exposed to. $Cd^{2+}$ suppressed growth to head-tail length at 0.1ppm. In conclusion, The study results reveal that $Cd^{2+}$ must be considered highly toxic effect to oocyte maturation and embryonic development.
Kim, Ae-Hyang;Kim, Yi-Soo;Piao, Zhe;Shin, Yong Chul;Ha, Min Woo
Korean Journal of Food Science and Technology
/
v.50
no.4
/
pp.420-429
/
2018
Skin ageing is associated with compromised performance of its fundamental barrier functions, with undesirable changes in appearance. Since this may introduce a detrimental impact on the quality of life, significant effort to discover effective ingredients against ageing is being invested. Recently, collagen hydrolysates containing tripeptides such as GlyPro-Hyp (GPH) have been developed with anticipation of improved effects compared to that of existing collagen hydrolysate-products. To evaluate the cutaneous hydration effect of collagen tripeptides (CTP), meaningful biomarkers in human dermal fibroblasts (HDF) and NC/Nga Tnd mice were analyzed in this study. Increased levels of ceramide kinase, hyaluronic acid, collagen 1A, and hyaluronan synthase-2 (HAS2), and decreased levels of hyaluronidase-1 (HYAL1) and CD44 in HDF cells were demonstrated. Furthermore, significant reduction of transepidermal water loss (TEWL), scratching behavior, HYAL1, $TNF-{\alpha}$ and IL-6 and increased water content and HAS2 were verified by in vivo tests. These results strongly suggest the potential of CTP as a skin hydration agent.
Cultured products (callus and exopolysaccharide) were obtained from suspension culture of Aloe vera callus, and the extracts of callus were further prepared with cold water or 60% ethanol solution. The ethanol extract of callus (AC) and exopolysaccharide (ACP) of 10 mg/mL exhibited the relatively higher suppression activity of 43.2-52.1% against hyaluronidase activity. Thus, their anti-inflammatory effects were further investigated using animal cell (Raw 264.7) in vitro. Though AC shows a slight suppression effect of cell survival rate (97%) using MTT assay in the presence of $400{\mu}g/mL$ AC- dimethyl sulfoxide (DMSO), cell growth promotion was observed in the other samples of lower levels. It indicates that the ethanol extract of Aloe callus rarely affect cell survival rate in the ranges ($200-400{\mu}g/mL$) used in the study. Using Griess reagent, the suppression of NO production by the aloe callus extract was analyzed by measuring the amount of the nitrite produced in Raw 264.7 culture activated by lipopolysaccharide (LPS). As a result, supplementation of AC-distilled water (DW) and AC-DMSO produced higher levels of NO than the positive control LPS. However, the NO suppression effect by ACP-DW was so intense that lower amount ($80-100{\mu}g/mL$) suppressed NO production to the level of the control. The effect was attributed to the expression of the iNOS. Then, Raw 264.7 cells were stimulated with the LPS and expression of COX-2 protein level was analyzed depending on the Aloe suspension culture product treatment. The results showed that the ACP-DW supplemented medium did not express COX-2 by itself, and LPS stimulated COX-2 expression was slightly decreased. On the other hand, realtime-PCR analysis of the expression of inflammatory cytokine showed that IL-$1{\beta}$ and TNF-${\alpha}$ expression was highly suppressed in the ACP- distilled water supplemented medium.
Park, So-Hyang;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Kim, Yong-Ki
Research in Plant Disease
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v.23
no.2
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pp.159-167
/
2017
This study was conducted to evaluate the control efficacy of the organic farming materials on cucumber scab caused by Cladosporium cucumerinum PT1 (KACC 48094). The antifungal activities in vitro as well as the suppressive effect of 43 organic farming materials on the spore germination and germ tube growth by inoculating spore suspension on cucumber seedlings in vivo were investigated. Thirteen organic farming materials inhibited the mycelial growth of C. cucumerinum and nine of these were microbial agents. In the screening using cucumber seedlings, six organic farming materials were very effective with control efficacy value of 90%. Among them, Bacillus amyloliquefaciens M27 provided suppressive effect on both mycelial growth and spore germination against cucumber scab. Finally, nine organic farming materials were selected to test the protective and curative effects, and all chosen organic farming materials significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, Bordeaux mixture I and III gave excellent protective control efficacy with control values of 96.7% and 73.3%, respectively, whereas its curative control effect was significant low. Among these, only Thymus quinquecostatus+Sophora extract showed curative activity, although the control value was as low as 50%. This study suggests that cucumber scab can be controlled by some organic farming materials in the farmhouses under comparatively cold and wet condition and protective treatment is more important and efficient.
Journal of the korean academy of Pediatric Dentistry
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v.31
no.4
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pp.555-563
/
2004
The pH of beverages is known to be low and have, therefore, been implicated in the increasing incidence of erosion. Erosion is believed to be the predominant cause of teeth wear in children and young adults, although there will always be a contribution from attrition and abrasion. The aim of the present study was to evaluate the effect of yogurt on the progression of erosive demineralization in human enamel using demineralization model in vitro. In 4 yogurts, available on the market, pH, buffering capacity and the concentrations of calcium, phosphate and fluoride were determined. The buffering effect was determined by titration with NaOH. 50 milliliters of each drink was then titrated with 1M sodium hydroxide, added in 0.5 milliliters increments, until the pH reached about 7. Human deciduous enamel(n=40) samples were divided into four groups and exposed to 80ml of the yogurt for 30,60, 90 and 120min. Enamel surface microhardness(VHN) was examined before and after each exposure. 1. The average PH of fermented milk was 3.77 and this pH value was acidic enough to cause tooth erosion. 2. All of the fermented milks were found to be erosive(p<0.05) 3. The teeth exposed to the fermented milk all showed erosion like lesions and microhardness measurements showed that enamel surface hardness decreased proportionately with increased time of immersion in all tooth specimen groups. 4. After immersion for 30 and 60 minutes, reduction rate of microhardness values was not significantly different between the groups(p>0.05). However, after 90 and 120 minutes, reduction rate of each group was significantly different(p<0.05).
Background: The leaves and roots of Panax ginseng are rich in ginsenosides. However, the chemical compositions of the leaves and roots of P. ginseng differ, resulting in different medicinal functions. In recent years, the aerial parts of members of the Panax genus have received great attention from natural product chemists as producers of bioactive ginsenosides. The aim of this study was the isolation and structural elucidation of novel, minor ginsenosides in the leaves of P. ginseng and evaluation of their antiinflammatory activity in vitro. Methods: Various chromatographic techniques were applied to obtain pure individual compounds, and their structures were determined by nuclear magnetic resonance and high-resolution mass spectrometry, as well as chemical methods. The antiinflammatory effect of the new compounds was evaluated on lipopolysaccharide-stimulated RAW 264.7 cells. Results and conclusions: Two novel, minor triterpenoid saponins, ginsenoside $LS_1$ (1) and 5,6-didehydroginsenoside $Rg_3$ (2), were isolated from the leaves of P. ginseng. The isolated compounds 1 and 2 were assayed for their inhibitory effect on nitric oxide production in LPS-stimulated RAW 264.7 cells, and Compound 2 showed a significant inhibitory effect with $IC_{50}$ of $37.38{\mu}M$ compared with that of NG-monomethyl-L-arginine ($IC_{50}=90.76{\mu}M$). Moreover, Compound 2 significantly decreased secretion of cytokines such as prostaglandin $E_2$ and tumor necrosis factor-${\alpha}$. In addition, Compound 2 significantly suppressed protein expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggested that Compound 2 could be used as a valuable candidate for medicinal use or functional food, and the mechanism is warranted for further exploration.
Purpose. This study aims to evaluate the combined effect of reduced thickness in different regions on the fracture resistance of monolithic zirconia crowns. Materials and methods. Seven nickel-chromium dies were generated from a 3D model of mandibular first molar using the digital scanner with the following geometries: 1.5 mm occlusal reduction, 1.0 mm deep chamfer. Based on the abutment model, Zirconia blocks (Luxen Zirconia) were selected to fabricate Sixty-three zirconia crowns with occlusal thicknesses of 0.3 mm, 0.5 mm, and 1.5 mm, and different axial thicknesses of 0.3 mm, 0.5 mm, and 1.0 mm. All crowns were cemented by resin cement. Next, the crowns were subjected to load-to-fracture test until fracture using an electronic universal testing machine. In addition, fracture patterns were observed with a scanning electron microscope (SEM). Two-way ANOVA and the Tuckey HSD test for post hoc analysis were used for statistical analysis (P < .05). Results. The mean values of fracture resistancerecorded was higher than the average biting force in the posterior region. The two-way ANOVA showed that the occlusal and axial thickness affected the fracture resistance significantly (P < .05). However, the effect of axial thickness on fracture resistance did not show a statistical difference when thicker than 0.5 mm. The observed failure modes were partial or complete fracture depending on the severity of crack propagation. Conclusion. Within the limitations of the present study, the CAD-CAM monolithic zirconia crown with extremely reduced thickness showed adequate fracture resistance to withstand occlusal load in molar regions. In addition, both occlusal and axial thickness affected the fracture resistance of the zirconia crown and showed different results as combined.
Total phenolics and flavonoids, and the antioxidant capacity of plum cultivar wines (Prunus salicina L. cv. Soldam and P. salicina L. cv. Formosa) were determined using spectrophotometric methods. The total phenolic and flavanoid contents of Soldam wine were $478.4\;{\pm}\;5.6\;mg$ GAE and $202.4\;{\pm}\;7.5\;mg$ CE per L,respectively, and in Formosa wine were $200.6\;{\pm}\;7.5\;mg$ GAE and $64.4\;{\pm}\;6.8\;mg$ CE per L, respectively. Neutral and acidic phenolics in Soldam wine were extracted with ethyl acetate and 0.01 N HCl, respectively. In the 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay, neutral phenolics (64.5 EDA%) had $3{\sim}4$ times higher antioxidant activity than acidic phenolics (21.5 EDA%) and other related phenolic compounds such as chlorogenic acid (15.5 EDA%) and quercetin (24.6 EDA%) at a concentration of $100\;{\mu}g/mL$. The antiviral activities of neutral and acidic phenolics in Soldam wine were investigated in vitro using a virus-induced cytopathic effect (CPE) inhibition assay. Results showed that neutral and acidic phenolics at concentrations of $100\;{\mu}g/mL$ inhibited porcine epidemic diarrhea virus (PEDV) replication at rates of 78.12% and 58.37%, respectively. The inhibition rate of 10 g/mL neutral phenolics (69.42%) was higher than that of ribavirin as an antiviral reagent (57.86%). At concentrations of $100\;{\mu}g/mL$ or less, neutral and acidic phenolics of Soldam wine had no cytotoxic effect against vero cells.
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