Park, Soon-Jung;Jeon, Young-Joo;Kim, Ju-Mi;Shin, Jeong-Min;Chae, Jung-Il;Chung, Hyung-Min
Reproductive and Developmental Biology
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v.34
no.3
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pp.143-151
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2010
Pluripotency of human embryonic stem cell (hESC) is one of the most valuable ability of hESCs for applying cell therapy field, but also showing side effect, for example teratoma formation. When transplant multipotent stem cell, such as mesnchymal stem cell (MSC) which retains similar differentiation ability, they do not form teratoma in vivo, but there exist limitation of cellular source supply. Accordingly, differentiation of hESC into MSC will be promising cellular source with strong points of both hESC and MSC line. In this study, we described the derivation of MSC like cell population from feeder free cultured hESC (hESC-MSC) using direct differentiation system. Cells population, hESC-MSC and bone marrow derived MSC (BM-MSC) retained similar characteristics in vitro, such as morphology, MSC specific marker expression and differentiation capacity. At the point of differentiation of both cell populations, differentiation rate was slower in hESC-MSC than BM-MSC. As these reason, to verify differentially expressed molecular condition of both cell population which bring out different differentiation rate, we compare the molecular condition of hESC-MSC and BM-MSC using 2-D proteomic analysis tool. In the proteomic analysis, we identified 49 differentially expressed proteins in hESC-MSC and BM-MSC, and they involved in different biological process such as positive regulation of molecular function, biological process, cellular metabolic process, nitrogen compound metabolic process, macromolecule metabolic process, metabolic process, molecular function, and positive regulation of molecular function and regulation of ubiquitin protein ligase activity during mitotic cell cycle, cellular response to stress, and RNA localization. As the related function of differentially expressed proteins, we sought to these proteins were key regulators which contribute to their differentiation rate, developmental process and cell proliferation. Our results suggest that the expressions of these proteins between the hESC-MSC and BM-MSC, could give to us further evidence for hESC differentiation into the mesenchymal stem cell is associated with a differentiation factor. As the initial step to understand fundamental difference of hESC-MSC and BM-MSC, we sought to investigate different protein expression profile. And the grafting of hESC differentiation into MSC and their comparative proteomic analysis will be positively contribute to cell therapy without cellular source limitation, also with exact background of their molecular condition.
Objective: The experiment aimed to determine the standardized ileal digestibility (SID) of crude protein (CP) and amino acids (AA) in 10 brown rice samples fed to pigs, and to construct predictive models for SID of CP and AA based on the physical characteristics and chemical composition of brown rice. Methods: Twenty-two cannulated pigs (initial body weight: 42.0±1.2 kg) were assigned to a replicated 11×3 incomplete Latin square design, including an N-free diet and 10 brown rice diets. Each period included 5 d adaptation and 2 d ileal digesta collection. Chromic oxide was added at 0.3% to all the diets as an indigestible marker for calculating the ileal CP and AA digestibility. Results: The coefficients of variation of all detected indices for physical characteristics and chemical composition, except for bulk weight, dry matter (DM) and gross energy, in 10 brown rice samples were greater than 10%. The SID of CP, lysine (Lys), methionine, threonine (Thr), and tryptophan (Trp) in brown rice was 77.2% (62.6% to 85.5%), 87.5% (80.3% to 94.3%), 89.2% (78.9% to 98.9%), 55.4% (46.1% to 67.6%) and 92.5% (86.3% to 96.3%), respectively. The best prediction equations for the SID of CP, Lys, Thr, and Trp were as following, SIDCP = -664.181+8.484×DM (R2 = 0.40), SIDLys = 53.126+6.031×ether extract (EE)+0.893×thousand-kernel volume (R2 = 0.66), SIDThr = 39.916+7.843×EE (R2 = 0.41), and SIDTrp = -361.588+4.891×DM+0.387×total starch (R2 = 0.85). Conclusion: Overall, a great variation exists among 10 sources of brown rice, and the thousand-grain volume, DM, EE, and total starch can be used as the key predictors for SID of CP and AA.
Lipids play many structural and metabolic roles, and dietary fat has great impact on metabolism and health. Fatty acid oxidation rate is dependent on tissue types. However there has been no report on the relationship between the rate of fatty acid oxidation and carnitine transport system in outer mitochondrial membrane of many tissues. In this study, the rate of fatty acid oxidation and carnitine palmitoyltransferase (CPT) I activity in the carnitine transport system were measured to understand the metabolic characteristics of fatty acid in various tissues. Palmitic acid oxidation rate and CPT I activity in various tissues were measured. Tissues were obtained from the white and red skeletal muscles, heart, liver, kidney and brain of rats. The highest lipid oxidation rate was demonstrated in the cardiac muscle, and the lowest oxidation rate was in brain. Red gastrocnemius muscle followed to the cardiac muscle. Lipid oxidation rates of kidney, white gastrocnemius muscle and liver were similar, ranging from 101 to 126 DPM/mg/hr. CPT I activity in the cardiac muscle was the highest, red gastrocnemius muscle followed by liver. Brain tissue showed the lowest CPT I activity as well as lipid oxidation rate, although the values were not significantly different from those of kidney and white gastrocnemius muscle. Therefore, lipid oxidation rate was highly (p<0.001) related to CPT I activity. Lipid oxidation rate is variable, depending on tissue types, and is highly (p<0.001) related to CPT I activity. CPT I activity may be a good marker to indicate lipid oxidation capacity in various tissues.
While much has been learned about the mechanisms of metastatic spread of cancer to bone, there has been little headway in establishing guidelines for monitoring the alteration in bone quality and estimating fracture risk. The aims of this study are, therefore, 1) to evaluate bone quality induced by metastatic bone tumor by analyzing the characteristics on bone microarchitecture and degree of bone mineralization and 2) analyze fracture risk increased secondary to the bone quality changes by metastatic bone tumor through calculating mechanical rigidities based on in-vivo micro CT images. For this study, eighteen female SD rats (12 weeks old, approximate 250 g) were randomly allocated in Sham and Tumor groups. W256 (Walker carcinosarcoma 256 malignant breast cancer cell) was inoculated in the right femur (intraosseous injection) in Tumor group, while 0.9% NaCl (saline solution) was injected in Sham group. The right hind limbs of all rats were scanned by in-vivo micro-CT to acquire structural parameters and degree of bone mineralization at 0 week, 4 weeks, 8 weeks, and 12 weeks after surgery. At the same time, urine was collected by metabolic cages for a biochemical marker test in order to evaluate bone resorption. Then, bone metastasis had been directly identified by positron emission tomography. Finally, axial, bending and torsional rigidities had been calculated based on in-vivo micro CT images for predict fracture risk. The results of this study showed that metastatic bone tumor might induce significant decrease in bone quality and increase of fracture risk. This study may be helpful to monitoring a degree of bone metastasis and predicting fracture risk due to metastatic bone tumor. In addition, this noninvasive diagnostic methodology may be utilized for evaluating other bone metabolic diseases such as osteoporosis.
Proceedings of the Korean Society of Applied Pharmacology
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2003.11a
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pp.79-79
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2003
Recently diabetes has been found to be associated with metabolic bone diseases such as osteoporosis. In the present study, attempts have been made-to explore the effect of high glucose in bone formation. Osteoblast-like UMR 106 cells were treated with high glucose (22mM, 33mM, 44mM) for 1 or 2 days. High glucose significantly inhibited proliferation of UMR106 cells in a time- and dose- dependent manner as evidenced by MTT assay. For the evaluation of collagen synthesis, UMR 106 cells were cultured in high glucose media (44mM) for 24 h and the ratio of collagen content to total protein was measured. In addition, gene expression pattern of type I collagen was assessed by RT-PCR. The high concentration of glucose inhibited a collagen synthesis, a marker of bone formation activity. JNK, c- Jun N-terminal Kinase, is known to play an important role in stress-associated cell death. In this regard, we tested to determine whether high glucose has any effect on JNK activity. It has been found that treatment of high glucose induced phosphorylation of JNK. On the other hand, ERK phosphorylation was inhibited by high glucose in a dose-dependent manner. Taken together, Therefore these results indicate that inhibition of proliferation in UMR 106 cells following high glucose is related to JNK/ERK containing signal pathways. This study showed high glucose concentration could alter the bone metabolism leading to defective bone formation, suggesting that high glucose due to diabetes may playa significant role in the development of metabolic bone disease.
In the present work , the transport mechanism of a capsaicin derivative, DA-5018, through blood-brain barrier (BBB) has been investigated to evaluate the feasibility of potential drug development. The result of pharmacokinetic parameters obtained from the intravenous injection of plasma volume marker,$[3^H]RSA$ and $[{14}^C]DA-5018$, indicated that both AUC, area under the plasma concentration curve and VD, volume of distribution in brain of $[3^H]RSA$ agreed with those reported ($1620{\pm}10 $percentage injected dose minute per milliliter (%IDmin/ml) and $12.0{\pm}0.1{\mu}l/g$, respectively). Elimination half-life and AUC of $[{14}^C]DA-5018$is corrected by the PHLC analysis, 19.6$\pm$1.2 min and 7.69$\pm$0.85% IDmin/ml, respectively. The metabolic rate of $[{14}^C]DA-5018$was very rapid. The blood-brain barrier permeability surface area (PS) product of $[{14}^C]DA-5018$ was calculated to be 0.24$\pm$0.05 $\mu$l/min/g. The result of internal carotid artery perfusion and capillary depletion suggested that [14C]DA-5018 pass through BBB with the time increasingly. Investigation of transport mechanism of $[{14}^C]DA-5018$ using agonist and antagonist suggested that vanilloid (capsaicin) receptor did not exist in the BBB, and nutrient carrier system in the BBB has no effect on the transport of DA-5018. In conclusion, despite the fact that penetration of DA-5018 through BBB is significant, the intact drug found in the brain tissue is small because of a rapid metabolism. Therefore, for the central analgesic effect of DA-5018, the method to increase the metabolic stability in plasma and the brain permeability should be considered.
A study was conducted to compare the endogenous basal losses of phosphorus (EBLP) in pigs fed diets containing gelatin (GEL) or spray-dried porcine plasma (SDPP) as protein sources and to determine the standardized total tract digestibility (STTD) of phosphorus (P) in SDPP. The trial was carried out at the Federal University of Santa Maria, Brazil. Twelve castrated pigs with an initial body weight of 55 kg were individually allotted in metabolic crates during two 12-day periods, each with 7 days of adaptation and 5 days of total fecal collection. The beginning and the end of the collecting periods were determined according to the marker-to-marker approach, using ferric oxide as an indigestible marker. Pigs were submitted to four semi-purified diets, one being a P-free diet with 30% of GEL as the protein source and three were diets with 10%, 20%, and 30% inclusion of SDPP respectively. Data were subjected to analysis of variance and the model included the effects of period, animal and treatments; the results of the three diets with increased levels of SDPP were subjected to linear regression analysis. The intercept of the relation of between ingested P and absorbed P represented the EBLP, while the slope indicated the STTD of P in SDPP. The EBLP means obtained by P-free diet and regression method were compared with the Student t test. The EBLP were 128.95 mg/kg dry matter intake (DMI) and 153.63 mg/kg DMI (standard error = 77.0; p<0.06) using the P-free diet with GEL as the protein source and the regression method, obtained with diets containing increased levels of SDPP, respectively. The apparent digestibility of P was 87.9%, 94.2%, and 92.9% for the treatments containing 10%, 20%, and 30% inclusion of SDPP, respectively. The estimated STTD of P obtained with the linear regression was 97.4%. When the EBLP estimated by the P-free diet was used to corrected the apparent digestibility of P in diets containing SDPP, the STTD of P in SDPP was 96.9%, 98.8%, and 95.9% for 10%, 20%, and 30% SDPP, respectively. Therefore, it can be concluded that SDPP can replace GEL to estimate the endogenous losses of P. In addition, the STTD of P in SDPP estimated with the P-free diet was 97.2% and it was 97.4% by the regression method, utilizing SDPP.
The capacity for liver regeneration involves a variety of nutritional factors. Vitamin C has multiple metabolic and antioxidant functions. In this study, we investigated the role of vitamin C in liver regeneration following hepatectomy in senescence marker protein (SMP)-30 knockout (KO) mice. Partial hepatectomy was performed by resecting the median and left lateral lobes of mice. Vitamin C accelerated liver recovery in SMP30 KO mice treated with vitamin C (KV). The livers of the KV mice exhibited lower levels of aspartate aminotransferase and lower injury than those of the KO mice. Increased type II transforming growth factor-β receptor (TGF-βRII)-mediated regeneration signaling was accompanied by HGF and cMet in the KV but not the KO mice. Consistent with this, the expression of cell cycle regulatory proteins, including cyclin D1 and proliferating cell nuclear antigen (PCNA), increased rapidly in the KV mice. Enhanced activation of ERK and GSK-3β proteins and a significantly increased number of binuclear hepatocytes were also detected in the livers of the KV mice. Moreover, the KV mice synthesized the highest levels of albumin. These data suggest that treating SMP30 knockout mice with vitamin C resulted in earlier recovery and liver regeneration by activation of the regeneration system.
The purpose of this study was to investigate the effects of resveratrol supplementation on inflammasome, inflammation, and macrophage markers in subcutaneous adipose tissue of high-fat-diet-induced obese mice. C57BL/6 mice were randomly assigned to three groups: normal diet control (NC; n=10), high-fat diet control (HC; n=10), or high fat with resveratrol (HRE; n=10) group. The mice were fed a high-fat diet (60% of calories from fat) or normal diet (18% of calories from fat). Resveratrol dissolved in a 0.1ml solution of dimethyl sulfoxide was supplemented orally at 25 mg/kg body weight. After 15 weeks, the body weight was significantly higher in the high-fat diet group than in the normal diet group. The inflammasome markers NLRP3, ASC, and caspase1 were significantly lower in the HRE group than in the HC group. The levels of an inflammation marker, IL-18, were also significantly lower in the HRE group than in the NC and HC groups. The levels of macrophage markers F480 and CD86 were significantly lower in the HRE group than in the HC group. The levels of the M2 macrophage marker CD206 were significantly decreased in the HC and HRE groups. Resveratrol had a positive effect on ameliorating the complications of high fat diet-induced obesity by reducing inflammasome and M1 macrophage gene expressions. However, resveratrol supplementation did not reduce inflammation gene expression.
Fernandez, Maria Luz;Jones, Jennifer J.;Ackerman, Daniela;Barona, Jacqueline;Calle, Mariana;Comperatore, Michael V.;Kim, Jung-Eun;Andersen, Catherine;Leite, Jose O.;Volek, Jeff S.;McIntosh, Mark;Kalynych, Colleen;Najm, Wadie;Lerman, Robert H.
Nutrition Research and Practice
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v.4
no.6
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pp.492-498
/
2010
Both metabolic syndrome (MetS) and elevated LDL cholesterol (LDL-C) increase the risk for cardiovascular disease (CVD). We hypothesized that low HDL cholesterol (HDL-C) would further increase CVD risk in women having both conditions. To assess this, we recruited 89 women with MetS (25-72 y) and LDL-C ${\geq}$ 2.6 mmol/L. To determine whether plasma HDL-C concentrations were associated with dietary components, circulating atherogenic particles, and other risk factors for CVD, we divided the subjects into two groups: high HDL-C (H-HDL) (${\geq}$ 1.3 mmol/L, n=32) and low HDL-C (L-HDL) (< 1.3 mmol/L, n=57). Plasma lipids, insulin, adiponectin, apolipoproteins, oxidized LDL, Lipoprotein(a), and lipoprotein size and subfractions were measured, and 3-d dietary records were used to assess macronutrient intake. Women with L-HDL had higher sugar intake and glycemic load (P< 0.05), higher plasma insulin (P< 0.01), lower adiponectin (P< 0.05), and higher numbers of atherogenic lipoproteins such as large VLDL (P < 0.01) and small LDL (P<0.001) than the H-HDL group. Women with L-HDL also had larger VLDL and both smaller LDL and HDL particle diameters (P<0.001). HDL-C was positively correlated with LDL size (r=0.691, P<0.0001) and HDL size (r=0.606, P<0.001), and inversely correlated with VLDL size (r=-0.327, P<0.01). We concluded that L-HDL could be used as a marker for increased numbers of circulating atherogenic lipoproteins as well as increased insulin resistance in women who are already at risk for CVD.
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