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Cholesterol-induced inflammation and macrophage accumulation in adipose tissue is reduced by a low carbohydrate diet in guinea pigs

  • Aguilar, David;deOgburn, Ryan C.;Volek, Jeff S.;Fernandez, Maria Luz
    • Nutrition Research and Practice
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    • v.8 no.6
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    • pp.625-631
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    • 2014
  • BACKGROUND/OBJECTIVES: The main objective of this study was to evaluate the effects of a high cholesterol (HC) dietary challenge on cholesterol tissue accumulation, inflammation, adipocyte differentiation, and macrophage infiltration in guinea pigs. A second objective was to assess whether macronutrient manipulation would reverse these metabolic alterations. MATERIALS/METHODS: Male Hartley guinea pigs (10/group) were assigned to either low cholesterol (LC) (0.04g/100g) or high cholesterol (HC) (0.25g/100g) diets for six weeks. For the second experiment, 20 guinea pigs were fed the HC diet for six weeks and then assigned to either a low carbohydrate (CHO) diet (L-CHO) (10% energy from CHO) or a high CHO diet (H-CHO) (54% CHO) for an additional six weeks. RESULTS: Higher concentrations of total (P < 0.005) and free (P < 0.05) cholesterol were observed in both adipose tissue and aortas of guinea pigs fed the HC compared to those in the LC group. In addition, higher concentrations of pro-inflammatory cytokines in the adipose tissue (P < 0.005) and lower concentrations of anti-inflammatory interleukin (IL)-10 were observed in the HC group (P < 0.05) compared to the LC group. Of particular interest, adipocytes in the HC group were smaller in size (P < 0.05) and showed increased macrophage infiltration compared to the LC group. When compared to the H-CHO group, lower concentrations of cholesterol in both adipose and aortas as well as lower concentrations of inflammatory cytokines in adipose tissue were observed in the L-CHO group (P < 0.05). In addition, guinea pigs fed the L-CHO exhibited larger adipose cells and lower macrophage infiltration compared to the H-CHO group. CONCLUSIONS: The results of this study strongly suggest that HC induces metabolic dysregulation associated with inflammation in adipose tissue and that L-CHO is more effective than H-CHO in attenuating these detrimental effects.

Insecticidal Activity and Histopathological Effects of Vip3Aa Protein from Bacillus thuringiensis on Spodoptera litura

  • Song, Feifei;Lin, Yunfeng;Chen, Chen;Shao, Ensi;Guan, Xiong;Huang, Zhipeng
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1774-1780
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    • 2016
  • Vegetative insecticidal proteins (Vips) are insecticidal proteins synthesized by Bacillus thuringiensis during the vegetative stage of growth. In this study, Vip3Aa protein, obtained by in vitro expression of the vip3Aa gene from B. thuringiensis WB5, displayed high insecticidal activity against Spodoptera litura aside from Spodoptera exigua and Helicoverpa armigera. Bioassay results showed that the toxicity of Vip3Aa protein against S. litura larvae statistically decreased along with the increase of the age of the larvae, with LC50 = 2.609 ng/cm2 for neonatal larvae, LC50 = 28.778 ng/cm2 for first instar larvae, LC50 = 70.460 ng/cm2 for second instar larvae, and LC50 = 200.627 ng/cm2 for third instar larvae. The accumulative mortality of 100% larvae appeared at 72 h for all instars of S. litura larvae, when feeding respectively with 83.22, 213.04, 341.40, and 613.20 ng/cm2 of Vip3Aa toxin to the neonatal and first to third instar larvae. The histopathological effects of Vip3Aa toxin on the midgut epithelial cells of S. litura larvae was also investigated. The TEM observations showed wide damage of the epithelial cell in the midgut of S. litura larvae fed with Vip3Aa toxin.

Tolerance and Histological Responses to Formalin of Black Seabream Acanthopagrus schlegelii Juveniles (감성돔(Acanthopagrus schlegelii) 치어의 포르말린에 대한 내성 및 조직학적 반응)

  • Myeong, Jeong-In;Min, Byung Hwa;Park, Mi Seon;Hwang, Hyung Kyu;Do, Jeung-Wan;Jeoung, Kyung Il;Chang, Young Jin;Jeong, Dal Sang
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.46 no.6
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    • pp.923-929
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    • 2013
  • Black seabream juveniles Acanthopagrus schlegelii held at $20^{\circ}C$ were exposed to formalin at concentrations of 50 to 400 ppm, and tolerance values were determined by calculating median lethal concentration values (LC50) through probit analysis. The 12-, 24, 48, 72- and 96-h LC50 values for formalin were 297, 233, 171, 162 and 157 ppm, respectively. The histological effects of formalin on gill and liver tissues in this fish were determined. No histological effects were observed in the control group. The intensity of cell damage increased with the concentration of, and duration of exposure to, formalin. Hyperplasia, separation and epithelial necrosis, epithelial lifting, lamellar synechiae and collapsed lamellae were observed in gill tissues exposed to formalin. Hepatic lesions in liver tissues of fishes exposed to formalin were characterized by cloudy swelling of hepatocytes, necrosis, cytoplasmic vacuolization, deposition of pigments, spongiosis hepatis, nuclear hypertrophy, dilation of sinusoids and bile stagnation. The LC50 values and histological results obtained in this study will aid in designing treatment regimens to minimize toxic side effects and increase efficacy.

Design of a Low-Pressure Hydrocyclone with Application for Fine Settleable Solid Removal Using Substitute Polystyrene Particles

  • Lee, Jin-Hwan;Jo, Jae-Yoon
    • Journal of Aquaculture
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    • v.18 no.3
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    • pp.189-195
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    • 2005
  • By testing the separation performance for a fine settleable solid removal system in an aquaculture system using polystyrene particles as an experimental substitute, the optimal geometric dimensions for a Low-Pressure Hydrocyclone (LPH) were obtained. The design approach far the LPH took into consideration two inflow diameters (Di: 30, 50 mm), three overflow diameters (Do: 60, 70, 100 mm) and four cylinder lengths (Lc: 250, 345, 442, 575 mm), while the cylinder diameter (Dc) at 335 mm, the underflow diameter (Du) at 50 mm and the cone angle (${\theta}$) at $68^{\circ}$ were kept constant. The separation performances of 19 different dimension combinations of LPH were tested, ranging from 300 to 1200 ml/sec of inflow rate using substitute polystyrene particles (0.4-0.7 mm dia., ${\rho}_s=1.05g/cm^3$). These polystyrene particles exhibit a similar density and settling velocity to the fine fecal debris of the common carp. The total separation efficiency for the inflow rate ranged from a high of 97% to a low of 20%. Experimental results obtained by ANCOVA and the Tukey test (${\alpha}=0.05$) showed that the separation performances of the LPH were significantly affected (P<0.05) by the fi, Di, Do and Lc. The maximum separation performance was detected at a dimension combination of 30 mm of inflow diameter (Di), 60 mm of overflow diameter (Do), 442 and 575 mm of cylinder length (Lc). The dimension proportions were 0.09, 1.32-1.72, 0.18 and 0.15 for Di/Dc, Lc/Dc, Do/Dc and Du/Dc respectively.

Induction of Conditioned Taste Aversion to Korean Pine Nuts (Pinus koraiensis) Treated with Lithium Chloride in Red Squirrels (Sciurus vulgaris)

  • Kim, Eui-Kyeong;Kim, Won-Myeong;Park, Yung-Chul;Yoo, Byung-Ho;Kim, Jong-Kuk
    • Journal of Ecology and Environment
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    • v.31 no.4
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    • pp.341-344
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    • 2008
  • We investigated the responses of red squirrels to pine nuts (Pinus koraiensis) treated with Lithium Chloride (LiCl) and the potential of the chemical for inducing conditioned taste aversion (CTA) in red squirrels. In red squirrels, nut feeding declined dramatically during the first 4 days after feeding with LiCl-treated nuts. The ratio of LiCl-treated nuts eaten to total nuts eaten declined from the $1^{st}$ day in LC-1 and the $2^{nd}$ day in LC-2, along with a general reduction in quantity eaten. Thus, feeding with LiCl-treated nuts induced CTA from the 2nd day after feeding, and CTA remained constant until the $4^{th}$ day, but disappeared on the $5^{th}$ day. The squirrels ate an average of $757.0{\pm}106.1mg$ (n = 2, range $682.0\sim832.0$) of LiCl before dying on the $16^{th}$ day of the study. The lethal dose of LiCl was 2.32 mg LiCl/g body weight, and the average amount of LiCl needed to induce CTA was $23.0{\pm}4.24mg$ (20 mg in LC-1 and 26 mg in LC-2).

A Study on the Production Medium of Lactic Acid Bacterial Cells by Using Corn Steep Liquor (Corm Steep Liquor를 이용한 젖산균이 생산배지에 관한 연구)

  • 안영태;김근배;인영민;정석근;함준상;김동운;이경욱;김선기;김현욱
    • Food Science of Animal Resources
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    • v.20 no.3
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    • pp.181-191
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    • 2000
  • 젖산균의 생장을 위한 질소, 탄소 공급원으로서 corn steep liquor의 이용 가능성을 시험하고 반응 표면 분석(Response surface metho-dology)을 이용하여 젖산균의 최적 생상 배지 조성을 연구하였다. 반응 표면 분석에서 L.fermentum의 생장배지에 첨가되 corn steep liquor와 yeast extract의 농도(p<0.01) 그리고 corn steep liquor와 yeast extract의 교호작용(P<0.05)이 L.ferm-entum의 생장에 큰 영향을 미치는 것으로 나타났으며, 이때 생균수가 최대인 corn steep liquor의 함량은 10.77%, yeast extract는 3.39%.Tween 80은 1.69%으로 예측되었다. 한편, Lc, lactis ssp. lactis 의 생장배지는 corn steep liquor 의 농도(P<0.01) 그리고 corn steep li-quor와 $\beta$-glycerophosphate disodium salt의 교호작용(P<0.05)인 Lc. lactis ssp. lactis의 생장에 큰 영향을 미치는 것으로 나타났으며 이때 생균수가 최대인 corn steep liquor의 함량은 3.5%, $\beta$-glycerophosphate disodium salt는 4.38%로 예측되었다. MRS broth와 예측된 최적 배지에서 L.fermentum의 젖산과 초산의 생성량은 각각 0.166, 0.114과 0.273, 0.081 M이고 M 17glc broth와 최적배지에서 Lc. lacti ssp. lactis의 젖산과 초산의 생성량은 각각 0.089, 0.003과 0.189, 0.003M이었다. 따라서 corn steep liquor는 L. fermentum와 Lc. lactis ssp, lactis 의 생장을 위해 질소 또는 탄소 공급원으로서 배지에 첨가 될 수 있는 우수한 농업 부산물로 판단되었다.

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MICROLEAKAGE AND MARGINAL ADAPTATION OF POSTERIOR COMPOSITE RESIN RESTORATIONS ACCORDING TO RESTORATIVE TECHNIQUE (수복에 따른 구치부 복합레진 수복물의 변연부 미세누출 및 적합도)

  • Yang, In-Seo;Shin, Dong-Hoo
    • Restorative Dentistry and Endodontics
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    • v.22 no.1
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    • pp.334-346
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    • 1997
  • Incidence of using esthetic composite resin in the posterior area is increasing but there were lots of inconsistent reports about their microleakage and marginal adaptation. The purpose of this study was to evaluate the differences of microleakage and marginal adaptation according to restorative techniques. 30 cavities with enamel gingival margin were prepared and restored with 3 types of composite resin [Z-100($Scotchbond^{TM}$ MP), AELITEFIL ($Onestep^{TM}$), Her culite XRV(Fuji BOND LC)] in direct technique and another 30 cavities were restored with preformed CR inlays and 3 different modern resin and resin-modified GI cements (Superbond C&B, Choice, Fuji Duet). Samples were chemically stressed in 75% ethanol for 24 hours and were thermocycled (5-$55^{\circ}C$(500 times. The degree of microleakage through proximal and gingival margins was examined by 1 % MB dye and the degree of marginal adaptation by examining the margins via SEM. The following results were obtained ; 1. In direct groups, Herculite XRV [Fuji BOND LC, 35.13 (15.50) %] group showed statistically different, less microleakage than Z-100 [$Scotchbond^{TM}$ MP, 72.91 (16.91 %] group and AELITEFIL [One-step, 93.73 (13.66) %] group (p<0.05). 2. In indirect groups, the degree of microleakage in Mean(S.D.) were: Super bond C & B [39.00 (24.35) %], Choice [57.19 (33.80) %], Fuji Duet [58.22 (40.36) %]. But there was no significant difference. 3. There was no significant difference between resin cement and resin-modified GI cement. 4. There were gaps at the interface with the tooth structure, but no gap was seen at the interface with restoration in all specimens. 5. In direct groups, Herculite XRV(Fuji BOND LC) group made little gap compared with other groups, but 40-$50{\mu}m$ thickness of bonding agent, Fuji BOND LC, looked like a cement used in indirect technique. 6. All indirect groups showed a variety of cement thickness, from less than $20{\mu}m$ to over $100{\mu}m$ and that dimension of buccal/lingual margin was less than that of gingival margin.

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Simultaneous Determination of Doxifluridine and 5-FU in Liver and Intestine Tissue Using LC/MS/MS (LC/MS/MS를 이용한 원숭이 및 비글견의 간 및 장관 조직에서의 Doxifluridine과 대사체 5-FU 동시분석법 개발)

  • Woo, Young-Ah;Kim, Ghee-Hwan;Jeong, Eun-Ju;Kim, Choong-Yong
    • YAKHAK HOEJI
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    • v.52 no.2
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    • pp.93-100
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    • 2008
  • A liquid chromatographic method with tandom spectrometric detection (LC/MS/MS) for the simultaneous determination of doxifluridine and its active metabolite, 5-fluorouracil (5-FU) was developed over the concentration range of $5{\sim}2000$ ng/ml, respectively. Doxifluridine, 5-FU and internal standard, 5-chlorouracil (5-CU), were extracted from liver and intestine tissue via protein precipitation. Acetonitrile was used as the extraction solvent and the supernatant was evaporated and reconstructed in mobile phase. Optimum chromatographic separation was achieved on a Agilent Zorbax $C_{18}$ ($100\;mm{\times}2.1\;mm$, $3.5\;{\mu}m$) column with mobile phase run in isocratic with methanol : water (20 : 80, v/v). The flow rate was 0.2 ml/min with total cycle time of 5 min. The lower limit of quantification was validated at 5.0 ng/ml of liver and intestine tissue, for both doxifluridine and 5-FU, respectively. The intra-day and inter-day precision and accuracy of quality control (QC) samples were <11% coefficient of variation and <7% relative error from theoretical concentration for both analytes. In addition, the special designed stability study was performed, because the metabolism of doxifluridine occurs spontaneously even in ice bath for monkey liver. The stability of doxifluridine in liver and intestine of monkey and beagle dog was compared. It was found that bioanalytical validation could not be performed for the monkey liver; however, beagle dog's liver has relatively low speed of metabolism compared to monkey liver and instead of monkey liver, beagle dog's liver could be used for the validation. Bioanalytical validation could be performed in monkey intestine. Eventually, this developed method for liver and intestine will be useful in support of the toxicokinetic and pharmacokinetic studies of doxifluridine and 5-FU.

Sensitive Determination of Pinaverium Bromide in Human Plasma by LC-ESI-MS/MS : Applicability to Oral Bioavailability Determination (LC-ESI-MS/MS를 이용한 생체시료 중 브롬화피나베리움의 고감도 분석 및 이를 이용한 생체이용률 평가)

  • Park, Seok;Lee, Ye-Rie;Kim, Ho-Hyun;Lee, Hee-Joo;Kim, Yoon-Gyoon;Youm, Jeong-Rok;Han, Sang-Beom
    • Journal of Pharmaceutical Investigation
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    • v.34 no.6
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    • pp.513-519
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    • 2004
  • A sensitive method for quantification of pinaverium bromide in human plasma was established using liquid chromatography-electrospray ionization tandem mass spectrometry(LC-ESI-MS/MS). Glimepiride was used as internal standard. Pinaverium bromide and internal standard in plasma sample were extracted using tert-butylmethylether(TBME). A centrifuged upper layer was then evaporated and reconstituted with mobile phase of acetonitrile-5 mM ammonium formate (80/20, pH 3.0). The reconstituted samples were injected into a $C_{18}$ reversed-phase column. Using MS/MS with multiple reaction monitoring (MRM) mode, pinaverium and glimepirde were detected without severe interference from human plasma matrix. Pinaverium produced a protonated precursor ion $([M+H]^+)$ at m/z 510.3 and a corresponding product ion at m/z 228.9. Internal standard produced a protonated precursor ion $([M+H]^+)$ at m/z 491.5 and a corresponding product ion at m/z 352.0. Detection of pinaverium bromide in human plasma was accurate and precise, with limit of quantitation at 0.5 ng/ml. The method has been successfully applied to bioavailability study of pinaverium bromide tablet in Korean healthy male volunteers. Pharmacokinetic parameters such as $AUC_t,\;C_{max},\;T_{max},\;K_{el}\;and\;t_{1/2}$ were calculated.

Analysis and Exposure Assessment of Perchlorate in Korean Dairy Products with LC-MS/MS

  • Oh, Sung-Hee;Lee, Ji-Woo;Mandy, Pawlas;Oh, Jeong-Eun
    • Environmental Analysis Health and Toxicology
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    • v.26
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    • pp.12.1-12.7
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    • 2011
  • Objectives: Perchlorate is an emerging contaminant that is found everywhere, including various foods. Perchlorate is known to disturb the production of thyroid hormones and leads to mental disorders in fetuses and infants, as well as metabolic problems in adults. In this study, we attempted to establish an LC-MS/MS method for measuring perchlorate in dairy products and used this developed method to investigate perchlorate levels in Korean milk and yogurt samples. Methods: The developed method of perchlorate analysis requires a shaker and 1% acetic acid/acetonitrile as the extracting solvent. Briefly, the samples were extracted and then centrifuged (4000 rpm, 1hour), and the supernatant was then passed through a $Envi^{TM}$ Carb SPE cartridge that had been prewashed sequentially with 6 mL of acetonitrile and 6 mL of 1% acetic acid in water. The final volume of the sample extract was adjusted to 40 mL with reagent water and the final sample was filtered through a 0.20-${\mu}m$ pore size PTFE (Polytetrafluoroethylene) syringe filter prior to LC-MS/MS. Results: The average levels of perchlorate in milk and yogurt samples were $5.63{\pm}3.49\;{\mu}g/L$ and $3.65{\pm}2.42\;{\mu}g/L$, respectively. The perchlorate levels observed in milk samples in this study were similar to those reported from China, Japan, and the United States. Conclusions: The exposure of Koreans to perchlorate through the consumption of dairy products was calculated based on the results of this study. For all age groups, the calculated exposure to perchlorate was below the reference of dose (0.7 ${\mu}g/kg$-day) proposed by the National Academy of Science, USA, but the perchlorate exposure of children was higher than that of adults. Therefore, further investigation of perchlorate in other food samples is needed to enable a more exact assessment of exposure of children to perchlorate.