The balance between bone formation by osteoblasts and destruction of mineralized bone matrix by osteoclasts is important for bone homeostasis. The increase of osteoclast differentiation by RANKL induces bone diseases such as osteoporosis. Recent studies have shown that insulin is one of main factors mediating the cross-talk between bone remodeling and energy metabolism. However, the systemic examination of insulin-induced differential gene expression profiles in osteoclasts has not been extensively studied. Here, we investigated the global effects of insulin on osteoclast precursors at the level of gene transcription by microarray analysis. The number of genes that were up-regulated by ${\geq}1.5$ fold after insulin treatment for 6 h, 12 h, or 24 h was 76, 73, and 39; and 96, 83, and 54 genes were down-regulated, respectively. The genes were classified by 20 biological processes or 24 molecular functions and the number of genes involved in 'development processes' and 'cell proliferation and differentiation' was 25 and 18, respectively, including Inhba, Socs, Plk3, Tnfsf4, and Plk1. The microarray results of these genes were verified by real-time RT-PCR analysis. We also compared the effects of insulin and RANKL on the expression of these genes. Most genes had a very similar pattern of expressions in insulin- and RANKL-treated cells. Interestingly, Tnfsf4 and Inhba genes were affected by insulin but not by RANKL. Taken together, these results suggest a potential role for insulin in osteoclast biology, thus contributing to the understanding of the pathogenesis and development of therapeutics for numerous bone and metabolic diseases.
Pseudomonas sp. K82 has been reported to be an aniline-assimilating soil bacterium. However, this strain can use not only aniline as a sole carbon and energy source, but can also utilize benzoate, p-hydroxybenzoate, and aniline analogues. The strain accomplishes this metabolic diversity by using dif-ferent aerobic pathways. Pseudomonas sp. K82, when cultured in p-hydroxybenzoate, showed extradiol cleavage activity of protocatechuate. In accordance with those findings, our study attempted the puri-fication of protocatechuate 4,5-dioxygenase (PCD 4,5). However the purified PCD 4,5 was found to be very unstable during purification. After Q-sepharose chromatography was performed, the crude enzyme activity was augmented by a factor of approximately 4.7. From the Q-sepharose fraction which exhibited PCD 4,5 activity, two subunits of PCD4,5 (${\alpha}$ subunit and ${\beta}$ subunit) were identified using the N-terminal amino acid sequences of 15 amino acid residues. These subunits were found to have more than 90% sequence homology with PmdA and PmdB of Comamonas testosteroni. The molecular weight of the native enzyme was estimated to be approximately 54 kDa, suggesting that PCD4,5 exists as a het-erodimer (${\alpha}$$_1$${\beta}$$_1$). PCD 4,5 exhibits stringent substrate specificity for protocatechuate and its optimal activity occurs at pH 9 and 15 $^{\circ}C$. PCR amplification of these two subunits of PCD4,5 revealed that the ${\alpha}$ subunit and ${\beta}$ subunit occurred in tandem. Our results suggest that Pseudomonas sp. K82 induced PCD 4,5 for the purpose of p-hydroxybenzoate degradation.
This study was performed to investigate the effects of recombinant BST formulation treatment on the milk yield, milk components, mastitis, and general cow health condition when the formulations of retinyl palmitate, cholecalciferol and rBST were administered after the peak period of milk production. The milk yields of treatment groups (Group I, II, III and IV) were increased from 21.5% to 29.0% than that of control group. There was significant difference in milk production between treatment group II, IV and control group (P<0.05). However, there was no significant difference in milk production between treatment group I, III and control group (P<0.05). And the addition of retinyl palmitate and cholecalciferol into rBST formulation did not increase the milk yield. The milk of treatment groups with sustained-release rBST did not show significant difference in milk components (milk fat, protein, lactose, and solid not fat). However, there were minor changes, primarily in fat content of milk, during the first few weeks of rBST administration. There was no incidence of clinical mastitis between rBST treatment groups and control group. Addition of high and medium concentration of retinyl palmitate and cholecalciferol into sustained-release rBST formulation was efficient in reduction of somatic cell count in milk. There was great energy deficit in all treatment groups compared with control group during the early study period. Thus, the body condition score of all treatment groups showed lower value than control group. No evidences of metabolic health problems, such as ketosis, milk fever, and downer cow were observed. Incidence of general lameness did not appear on all treatment groups during 140 days of this study.
Carnitine is known to be involved in lipid metabolism and affects body composition as well as energy metabolism of the whole body. Improvement of obesity by L-carnitine supplement suggests that obesity can be related with the abnormality of carnitine metabolism and therefore, plasma carnitine level in normal and obesity groups was investigated. For the characterization of plasma carnitine level in obese people, 60 plasma samples collected from Korean women subjects were analyzed using LC/MS and plasma fatty acid level was also determined using GC/MS. Additionally, several clinical chemical parameters including fasting glucose, cholesterol, AST, and ALT level were measured. All the data obtained were combined and pattern recognition analysis was carried out with the dataset. Obese group showed a different metabolic pattern compared with normal group. Plasma acylcarnitine level of the obese group was found to be $11.7{\mu}g/ml$, which was higher than that of normal group ($8.0{\mu}g/ml$). Statistically significant differences in plasma fatty acid level were not observed between the two groups. Other clinical parameters for the obese group were within normal ranges but AST and ALT levels were slightly elevated compared to normal group. The obese group showed elevated plasma acylcarnitine level.
ATP synthase produces ATP, a major energy source for metabolic processes in organisms, from ADP and inorganic phosphate in cellular membranes. ATP synthase is known as a rotary motor, in which the c-subunit ring functions as a rotor. In this work, we have tried to develop a more general preparation procedure of thermophilic $F_oc$-ring ($TF_oc$-ring) for NMR measurements. The expression of $TF_oF_1$ is easily affected by various experimental conditions such as temperature, shape and size of a flask, a volume of medium, and shaking rate of an incubator. Accordingly, we have tried to optimize the expression conditions of $TF_oF_1$. $TF_oc$-rings were purified from $TF_oF_1$ according to a reported method. We modified purification procedures to improve purity and yield of $TF_oc$. On top of them, we found a new combination of detergents for the purification at anion-exchange column chromatography. To examine the effect of lipid environments on the structure, the $TF_oc$-rings were reconstituted into two kinds of lipid bilayers, namely, saturated and unsaturated lipid ones. Then, we have compared characteristics of the $TF_oc$-ring structures in these membranes with solid-state NMR.
This study was designed to describe the effect of the protein quality at different intake level of protein on the protein metabolism in the whole body of growing pigs with a simulation model. Varying to the protein level in feeds, four simulations were conducted. The feed protein level, represented as proportions of digestible protein to the metabolic energy (DP/ME, g/MJ), were 6-8, 11-13, 17-19, and 23-25 DP/ME, respectively. Two protein quality and six weeks of growth time were used at each simulation. The objective function for the simulations was protein deposition in the whole body, which was calculated from the experimental results. The parameters in the simulation were determined by the parameter estimation technique. The results obtained from the simulation were as follows: The protein synthesis and breakdown rates(g/day) in the whole body was increased with the increase of protein quality only at lower or required level of protein intake. They showed a parallel behavior in the course of growth, irrespective of quality and level of feed protein intake. The simulated protein deposition and protein synthesis showed a linear relationship between them at different protein quality and level. The affinity parameter showed a linear relationship between them at different protein quality and level. The affinity parameter showed that arginine, tryptophan and isoleucine were more efficient in the stimulation ofbody protein synthesis. Lysine and phenylalanine+tyrosine were less efficient. The oxidation parameter showed that histidine, pheyalanine+tyrosine were less efficient. The oxidation parameter showed that histidine, phenyalanine+tyrosine, and methionine+cystine were oxidized in larger magnitude than lysine and threonine. The oxidation parameter of most amino acids increased with the increase of protein intake beyond the requirement level, but not any more at highest protein intake level. Finally it was found that the improvement of feed protein quality at the lower or required level of protein intake increase protein deposition through a parallel increase of protein synthesis and breakdown.
Park, Jungwook;Lee, Areum;Lee, Hyun-Hee;Park, Inmyoung;Seo, Young-Su;Cha, Jaeho
Genes and Genomics
/
v.40
no.11
/
pp.1157-1167
/
2018
Sulfolobus species can grow on a variety of organic compounds as carbon and energy sources. These species degrade glucose to pyruvate by the modified branched Entner-Doudoroff pathway. We attempted to determine the differentially expressed genes (DEGs) under sugar-limited and sugar-rich conditions. RNA sequencing (RNA-seq) was used to quantify the expression of the genes and identify those DEGs between the S. acidocaldarius cells grown under sugar-rich (YT with glucose) and sugar-limited (YT only) conditions. The functions and pathways of the DEGs were examined using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Quantitative real-time PCR (qRT-PCR) was performed to validate the DEGs. Transcriptome analysis of the DSM 639 strain grown on sugar-limited and sugar-rich media revealed that 853 genes were differentially expressed, among which 481 were upregulated and 372 were downregulated under the glucose-supplemented condition. In particular, 70 genes showed significant changes in expression levels of ${\geq}$ twofold. GO and KEGG enrichment analyses revealed that the genes encoding components of central carbon metabolism, the respiratory chain, and protein and amino acid biosynthetic machinery were upregulated under the glucose condition. RNA-seq and qRT-PCR analyses indicated that the sulfur assimilation genes (Saci_2197-2204) including phosphoadenosine phosphosulfate reductase and sulfite reductase were significantly upregulated in the presence of glucose. The present study revealed metabolic networks in S. acidocaldarius that are induced in a glucose-dependent manner, improving our understanding of biomass production under sugar-rich conditions.
Dietary fiber is defined as soluble and insoluble polysaccharide consisted in the plant cell wall-associated fibers naturally occurring in fruits, vegetables, and cereal products, and of isolated fibers that are added to processed foods which are also artificially modified. There are so many difference types of dietary fibers as arabinoxylan, polydextrose chicory, oligosccharide. inulin, pectin, bran, cellulose, ${\beta}$-glucan, resistant starch and some seaweed polymers as alginate. Most of them provide many biological benefits in the intestine, as lower risk for developing coronary heart disease, stroke, hypertension, diabetes, obesity and some of the gastrointestinal disease like as colon cancer. And also lowering cholesterol levels, improves glycemic and insulin sensitivity to non-diabetic and diabetic persons including immune system. Beside of many benefits, average consumers in developed and under developing countries take far less amounts of dietary fiber that international organization recommended. Adequate intake of dietary fiber is 14g/1,000kcal base using the energy guide line of 2,000kcal/day for women and 26,000 kcal/day for men, dietary intake is 28g/day of adult women and 36g/day for adult men. The mechanisms behind the reported effects of dietary fiber on metabolic health are not fully well established. It is suggested that changes in intestinal viscosity resulting mucus increasing, macro-nutrients absorption, rate of passage of large intestinal, production of short chain fatty acids by fermentation. Production of gut hormones and changes of microbiota in intestine. It is necessary to do more research in this field in the future and combined interdisciplinary works together.
Objective: Intramuscular fat (IMF) content plays an important role in meat quality. Identification of single nucleotide polymorphisms (SNPs) and genes related to pig IMF, especially using pig populations with high IMF content variation, can help to establish novel molecular breeding tools for optimizing IMF in pork and unveil the mechanisms that underlie fat metabolism. Methods: We collected muscle samples of 453 Chinese Lulai black pigs, measured IMF content by Soxhlet petroleum-ether extraction method, and genotyped genome-wide SNPs using GeneSeek Genomic Profiler Porcine HD BeadChip. Then a genome-wide association study was performed using a linear mixed model implemented in the GEMMA software. Results: A total of 43 SNPs were identified to be significantly associated with IMF content by the cutoff p<0.001. Among these significant SNPs, the greatest number of SNPs (n = 19) were detected on Chr.9, and two linkage disequilibrium blocks were formed among them. Additionally, 17 significant SNPs are mapped to previously reported quantitative trait loci (QTLs) of IMF and confirmed previous QTLs studies. Forty-two annotated genes centering these significant SNPs were obtained from Ensembl database. Overrepresentation test of pathways and gene ontology (GO) terms revealed some enriched reactome pathways and GO terms, which mainly involved regulation of basic material transport, energy metabolic process and signaling pathway. Conclusion: These findings improve our understanding of the genetic architecture of IMF content in pork and facilitate the follow-up study of fine-mapping genes that influence fat deposition in muscle.
As recently many studies on the relationship between excessive sugar intake and chronic disease have been reported across the world, more attention to sugar intake has been paid. Sugars, called simple sugars, is a kind of carbohydrate and sum of monosaccharides and disaccharides. Sugars that are particularly related to health are added sugars or free sugars, which are added to food by processing or cooking. It is generally not to give satiety and increases energy density without other nutrients. Especially, the major food source of added sugar is sugar-sweetened beverages(SSBs), from which sugars are closely linked with health outcome. The total sugar intake in Korea is 61.4 g per day and it is almost half of those in the western countries such as the U.S. or Australia and it is within the recommendation level by the Dietary Reference Intakes for Koreans. However, when it is classified by age groups or food sources, sugar intake in adolescents and young adults are high and the sugar from processed foods is also high that would lead to high intake levels of added sugar. Particularly, the frequency of SSBs, the major food source of added sugar, is the highest among 20's. While the studies of excessive sugar intake with chronic diseases including obesity, type 2 diabetes, metabolic disease, cardiovascular disease reported in the Western populations, those in the Korean populations have just started. Given our dietary practice is very different from those in the western population, more prospective studies would be necessary to evaluate sugar intake by type or food source exactly and examine its relationship with chronic diseases in the Korean population.
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