The precise effects of protein intake on fractional synthesis rate (FSR) of muscle protein are still under debate. The sample size of these studies was small and the conclusions in young and elderly subjects were inconsistent. To assess the effect of dietary protein intake on the FSR level, we conducted a meta-analysis of controlled protein intake trials. Random-effects models were used to calculate the weighted mean differences (WMDs). Ten studies were included and effects of short-term protein intake were evaluated. In an overall pooled estimate, protein intake significantly increased the FSR (20 trials, 368 participants; WMD: 0.025%/h; 95%CI: 0.019-0.031; P < 0.0001). Meta-regression analysis suggested that the protein dose was positively related to the effect size (regression coefficient = 0.108%/h; 95%CI: 0.035, 0.182; P = 0.009). A subgroup analysis indicated that protein intake significantly increased FSR when the protein dose was ${\leq}$ 0.80 g/kg BW (16 trials, 308 participants; WMD: 0.027%/h; 95%CI: 0.019-0.031; P < 0.0001), but did not affect FSR when the protein dose was > 0.80 g/kg BW (4 trials, 60 participants; WMD: 0.016%/h; 95%CI: 0.004-0.029; P = 0.98). In conclusion, this study is the first integrated results showing that a short-term protein intake is effective at improving the FSR of muscle protein in the healthy elderly as well as young subjects. This beneficial effect seems to be dose-dependent when the dose levels of protein range from 0.08 to 0.80 g/kg BW.
Endocytosis is a process by which cells absorb extracellular materials via the inward budding of vesicles formed from the plasma membrane. Receptor-mediated endocytosis is a highly selective process where receptors with specific binding sites for extracellular molecules internalize via vesicles. G protein-coupled receptors (GPCRs) are the largest single family of plasma-membrane receptors with more than 1000 family members. But the molecular mechanisms involved in the regulation of GPCRs are believed to be highly conserved. For example, receptor phosphorylation in collaboration with ${\beta}$-arrestins plays major roles in desensitization and endocytosis of most GPCRs. Nevertheless, a number of subsequent studies showed that GPCR regulation, such as that by endocytosis, occurs through various pathways with a multitude of cellular components and processes. This review focused on i) functional interactions between homologous and heterologous pathways, ii) methodologies applied for determining receptor endocytosis, iii) experimental tools to determine specific endocytic routes, iv) roles of small guanosine triphosphate-binding proteins in GPCR endocytosis, and v) role of post-translational modification of the receptors in endocytosis.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.30
no.3
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pp.186-192
/
2004
Purpose: Human tooth proteins are highly heterogeneous, comprising diverse proteins derived from a number of genes. The attempts to identify protein for activity of tooth matrix proteins have been defied by several factors. First, the amount of proteins within teeth is very small relative to many extracellular matrix proteins of other tissues. Second, the bioassay system is tedious and needed for long time. Therefore we tried to find easy techniques, which increase the product rate, and an assay of small proteins, with which amino acid sequence is possible without additional procedures. Materials and Methods: Total protein were extracted from 300 g enamel removed teeth and 600 g teeth with 4 mol/L guanidine HCl and purified by gel chromatography. Aliquot of proteins was implanted into muscle pouches in Sprague-Dawley rats for bioassay. By SDS-PAGE and membrane blotting, molecular weight of each protein was estimated and a partial amino acid sequence was obtained. Each fraction blotted on the membrane was cut out and inserted in rat ectopic model. Results: In dissociative method, total tooth proteins were obtained 1mg/ml from enamel removed teeth and 3.5 mg/ml from teeth. In SDS-PAGE, four clear bands at the sites corresponding to 66, 40, 20 and 18 kD. Especially The 66 kD band was clearly exhibited. Amino acid sequencing from tooth could be possible using PVDF membrane blotting technique. In amino acid sequencing, 66 kD protein was identified as albumin. Conclusion: Compared with conventional method for extraction of teeth protein and bioassay of proteins, the methods in this study were easy, time-saving and more productive technique. The matured tooth proteins omitting additional procedure of mechanical removal of enamel were simply analyzed using blotted PVDF membrane. This method seems to make a contribution as a technique for bioassay and amino acid sequencing of protein.
The effects of chromium (Cr), dietary crude protein (CP) level, and potential interactions of these two factors were investigated in term of energy metabolism in lambs. Forty-eight 9-week-old weaned lambs (Dorper${\times}$Small-tail Han sheep, male, mean initial body weight = 22.96 kg${\pm}$2.60 kg) were used in a 2${\times}$3 factorial arrangement of supplemental Cr (0 ${\mu}g$/kg, 400 $\mu{g}$/kg or 800 ${\mu}g$/kg from chromium yeast) and protein levels (low protein: 157 g/d to 171 g/d for each animal, or high protein: 189 g/d to 209 g/d for each animal). Blood samples were collected at the beginning and end of the feeding trial. The lambs were then sacrificed and tissue samples were frozen for further analysis. Chromium at 400 ${\mu}g$/kg decreased fasting insulin level and the ratio of plasma insulin to glucagon, but these differences were not statistically significant; in contrast, chromium at 800 ${\mu}g$/kg increased the ratio significantly (p<0.05). Protein at the high level increased plasma tumor necrosis factor $\alpha$ (TNF-$\alpha$) level (p = 0.060). Liver glycogen content was increased significantly by Cr (p<0.05), which also increased liver glucose-6-phosphatase (G-6-Pase) and adipose hormone-sensitive lipase (HSL) activity. At 400 ${\mu}g$/kg, Cr increased muscle hexokinase (HK) activity. High protein significantly increased G-6-Pase activities in both the liver (p<0.05) and the kidney (p<0.05), but significantly decreased fatty acid synthase (FAS) activity in subcutaneous adipose tissue (p<0.05). For HSL activity in adipose tissue, a Cr${\times}$CP interaction (p<0.05) was observed. Overall, Cr improved energy metabolism, primarily by promoting the glycolytic rate and lipolytic processes, and these regulations were implemented mainly through the modulation by Cr of the insulin signal transduction system. High protein improved gluconeogenesis in both liver and kidney. The interaction of Cr${\times}$CP indicated that 400 $\mu{g}$/kg Cr could reduce energy consumption in situations where energy was being conserved, but could improve energy utilization when metabolic rate was increased.
An OTHBVS cell line from HepG2 was established. This cell line stably expresses the human hepatitis B virus (HBV) middle S protein that includes the preS2 region which is important for HBV particle entry into the hepatocyte. To establish this cell line, the middle S open reading frame (ORF), with a promoter located in the 5' region and enhancer located in the 3' region, was cloned downstream from the metallothionine (MT) promoter of the OT1529 vector. In this vector, expression of the middle S protein was constructed to be regulated by its own promoter and enhancer. Expression of the large S protein which contains the preS1 region in addition to the middle S protein was designed to be regulated by the MT promoter. When extracts of OTHBVS cells were examined with an S protein detection kit (RPHA, Korea Green Cross Co.), an S protein was detected. Total mRNA of OTHBVS cell examined by northern blot analysis with an S ORF probe revealed small/middle S transcripts (2.1 kb). When the MT promoter was induced by Zn, large S transcripts (2.4 kb) were detected. The GP36 and GP33 middle S proteins were presumably detected, but large S proteins were not detected by immunostain analysis using anti-preS2 antibody.
Jung, Samooel;Bae, Young Sik;Yong, Hae In;Lee, Hyun Jung;Seo, Dong Won;Park, Hee Bok;Lee, Jun Heon;Jo, Cheorun
Asian-Australasian Journal of Animal Sciences
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v.28
no.12
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pp.1760-1766
/
2015
This study investigated the proximate composition and $\small{L}$-carnitine and betaine content of meats from 5 lines of Korean indigenous chicken (KIC) for developing highly nutritious meat breeds with health benefits from the bioactive compounds such as $\small{L}$-carnitine and betaine in meat. In addition, the relevance of gender (male and female) and meat type (breast and thigh meat) was examined. A total of 595 F1 progeny (black [B], grey-brown [G], red-brown [R], white [W], and yellow-brown [Y]) from 70 full-sib families were used. The moisture, protein, fat, and ash contents of the meats were significantly affected by line, gender, and meat type (p<0.05). The males in line G and females in line B showed the highest protein and the lowest fat content of the meats. $\small{L}$-carnitine and betaine content showed effects of meat type, line, and gender (p<0.05). The highest $\small{L}$-carnitine content was found in breast and thigh meats from line Y in both genders. The breast meat from line G and the thigh meat from line R had the highest betaine content in males. The female breast and thigh meats showed the highest betaine content in line R. These data could be valuable for establishing selection strategies for developing highly nutritious chicken meat breeds in Korea.
Changes in free amino acid concentrations is blood and various tissues were evaluated in cats adapted to the low-protein diet(20% protein, LPD) or the high-protein diet(60% protein, HPD) for 5 weeks. Cumulative body weigth gain for the 5 week period was 463$\pm$43g, and -128$\pm$40g for cats fed HPD and LPD, respectively. Feeding HPD significantly increased the size of liver and kidney. Cats adapted to HPD for 5 weeks have significantly elevated plasma concrntrations of essential amino acids (branched-chain amino acides, threonine, trytophan, phenylalanine and methoionine), whereas plasma levels of non-essential amino acids(alanine, asparagine, glycine, glutamine and serine) were significantly reduced in animals adapted to HPD(p<0.01, or p<0.001) compared to the values for the cats fed LPD. Changes in free amino acid concentratioks in whole blood induced by the variations in dietary level of protein closely reflect the pattern seen in plasma. Amino acids such as branched-chain amino acids, proline and threonine were most difficult to maintain homeostasis and consistantly elevated in lever, kidney, skeletal muscle and brain, as well as in blood of cats adapted to HPD(p<0.01 or p<0.001). All of the free amino acids in jejunum, excluding taurine and ornithine, were significantly elevated in animals adapted to HPD, most probably due to the rapid absorption of large amount of amino acids across the epithelium of small intestine.
Previous studies have demonstrated that oxidative stress involving generation of reactive oxygen species (ROS) is responsible for the cytotoxic action of $TNF{\alpha}$. Protective effect of small heat shock proteins (small HSP) against diverse oxidative stress conditions has been suggeted. Although overexpression of small hsp was shown to provide an enhanced survival of $TNF{\alpha}$-sensitive cells when challenged with $TNF{\alpha}$, neither the nature of $TNF{\alpha}$-induced cytotoxicity nor the protective mechanism of small HSP has not been completely understood. In this study, we have attempted to determine whether $TNF{\alpha}$ induces oxidative DNA damage in $TNF{\alpha}$-sensitive L929 cells. We chose to measure the level of 8-hydroxy-2'-deoxyguanosine (8 ohdG), which has been increasingly recognized as one of the most sensitive markers of oxidative DNA damage. Our results clearly demonstrated that the level of 8 ohdG increased in L929 cells in a $TNF{\alpha}$ dose-dependent manner. Subsequently, we asked whether small HSP has a protective effect on $TNF{\alpha}$-induced oxidative DNA damage. To accomplish this goal, we have stably transfected L929 cells with mouse small hsp cDNA (hsp25) since these cells are devoid of endogenous small hsps. We found that $TNF{\alpha}$-induced 8 ohdG was decreased in cells overexpressing exogenous small hsp. We also found that the cell killing activity of $TNF{\alpha}$ was decreased in these cells as measured by clonogenic survival. Taken together, results from the current study show that cytotoxic mechanism of $TNF{\alpha}$ involves oxidative damage of DNA and that overexpression of the small hsp reduces this oxidative damage. We suggest that the reduction of oxidative DNA damage is one of the most important protective mechanisms of small HSP against $TNF{\alpha}$.
This study aimed to verify the nutritional and curative effects of protein hydrolysate in rats with cysteamine-induced duodenal uncer. Duodenal ulcer rat model was established by intraperitoneal injections of cysteamine. Sprague-Dawley, female rats weighing approximately 200g were intraperitoneally injected twice cysteamine(13mg/100g BW) at intervals of 3h per day. This procedure was repeated 3$\times$at intervals of 3d. Animals fed on 10% casein diet for infection periods. After last injection, 4 kinds of diets(10% casein, 20% casein, 10% casein hydrolysate, 20% casein hydrolysate) were given. Gastric montility, trypsin activity in gastrointestinal content, retention rate of nitrogen, plasma total protein, albumin, amino-N, urinary urea nitrogen, creatinine and hydroxyproline were analyzed for nutritional effects of dietary nitrogen levels(10%, 20%) and sources(casein, casein hydrolysate). In duodenal ulcer rat model, there was no differences between 20% casein diet and 20% casein hydrolysate in the view of severeness of ulcer, gastric emptying rate, serum total protein, serum albumin, plasma $\alpha$-amino-N, UUN, creatinine excretion, GFR, nitrogen retention. On the other hand, rats on 10% casein hydrolysate diet group had more curative effect of the ulcer, higher plasma albumin concentration and nitrogen retention than 10% casein diet group. The casein hydrolysate diet group was lower trypsin activity in small intestinal content than the casein diet group, at both nitrogen levels(10%, 20%). The results suggest that protein hydrolysate be applied in diet therapy for the patients with gastrointestinal ulcer.
The effect of undecorticated sunflower cake (USFC) as critical protein supplement was assessed and compared with deoiled groundnut cake (DGNC) in adult goats and sheep. The animals were fed a basal diet of wheat straw ad libitum and supplemented with either USFC or DGNC to meet their protein requirement for maintenance. Total dry-matter intake by sheep and goats (g/kg $BW^{0.75}$) on USFC was similar to their counterparts on DGNC supplemented group. However, while intake of cake moiety was significantly (p<0.05) higher in USFC, the intake of wheat straw was significantly (p<0.05) higher by animals on DGNC. Digestibility of various nutrients, except lower crude protein digestibility by goats in USFC group, did not differ significantly between animals given DGNC or USFC. DCP and TDN concentration (% DM) was comparable in sheep and goats irrespective of dietary supplement. Similarly, the intake (g/kg $W^{0.75}$) of DCP, DDM, DOM, and TDN was similar between DGNC and USFC in both sheep and goats. It may be concluded that undecorticated sunflower cake is comparable to deoiled groundnut cake as a critical protein supplement to the roughage based diet of small ruminants.
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