Proceedings of the Plant Resources Society of Korea Conference
/
2010.05a
/
pp.14-14
/
2010
Plastid proteomics are essential organelles present in virtually all cells in plants and green algae. Plastids are responsible for the synthesis and storage of key molecules required for the basic architecture and functions of plant cells. The proteome of plastid, and in particular of chloroplast, have received significant amounts of attention in recent years. Various fractionation and mass spectrometry (MS) techniques have been applied to catalogue the chloroplast proteome and its sub-organelles compartments. To better understanding the function of the lumenal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine-SDS-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, TMHMM, and TOPPRED) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.
The degree of attraction of Toxoplasma gondii to vertebrate cells varies with cell type and cell phase. Human promyelocytic leukemia cells, HL-60, were synchronized by double thymidine block method and co-cultured with Toxoplasma for 1 hr at each cell stage to investigate the cell cycle specific susceptibility of parasites to host cells. For 30 hr the average number of Texoplasma that invaded was a little changed except at 3 hr from G1/S phase boundary which concurred with the peak point of DNA synthesis. At 3 hr which is a relatively short interval compared to whole S phase, modification of cells by parasitic invasion was most remarkable. The number of Toxoplasma that penetrated was increased to more than sin times. The shape of the cells became sludgy and almost indiscernible by strong accessibility of parasites only for an hour of mfd-S phase. The same auctuation was also observed at the second peak of S phase but weakly. This suggests that there be surface molecules concerning with the attachment of Texoplasma to the host cells, which is expressed at special point of S phase. further studies on the specific protein or similar molecules related could be carried out using synchronized HL-60 cells.
Atherosclerosis and post-angiography restenosis are associated with intimal thickening and concomitant vascular smooth muscle cell (VSMC) proliferation. Obovatol, a major biphenolic component isolated from the Magnolia obovata leaf, is known to have anti-inflammatory and anti-tumor activities. The goal of the present study was to enhance the inhibitory effects of obovatol to improve its potential as a preventive or therapeutic agent in atherosclerosis and restenosis. Platelet-derived growth factor (PDGF)-BB-induced proliferation of rat aortic smooth muscle cells (RASMCs) was examined in the presence or absence of a newly synthesized obovatol derivative, OD78. The observed anti-proliferative effect of OD78 was further investigated by cell counting and [$^3H$]-thymidine incorporation assays. Treatment with 1-4 ${\mu}M$ OD78 dose-dependently inhibited the proliferation and DNA synthesis of 25 ng/ml PDGF-BB-stimulated RASMCs. Accordingly, OD78 blocked PDGF-BB-induced progression from the $G_0/G_1$ to S phase of the cell cycle in synchronized cells. OD78 decreased the expression levels of CDK4, cyclin E, and cyclin D1 proteins, as well as the phosphorylation of retinoblastoma protein and proliferating cell nuclear antigen; however, it did not change the CDK2 expression level. In addition, OD78 inhibited downregulation of the cyclin-dependent kinase inhibitor (CKI) $p27^{kip1}$. However, OD78 did not affect the CKI $p21^{cip1}$ or phosphorylation of early PDGF signaling pathway. These results suggest that OD78 may inhibit PDGF-BB-induced RASMC proliferation by perturbing cell cycle progression, potentially through $p27^{kip1}$ pathway activation. Consequently, OD78 may be developed as a potential anti-proliferative agent for the treatment of atherosclerosis and angioplasty restenosis.
Four Thai - rumen fistulated male swamp buffaloes (Bubalus bubalis), about four years old with $400{\pm}20kg$ liveweight, were randomly assigned according to a $4{\times}4$ Latin square design to receive dietary treatments. The treatments were: ground corn cob (GCC) replacement for cassava chip (CC) in concentrate at 0% (T1); GCC replacement at 33% (T2); GCC replacement at 67% (T3); and GCC replacement at 100% (T4), respectively. During the experiment, concentrate was offered at 0.5% BW while 5% urea-treated rice straw was given at ad libitum. The result revealed that there was no effect of GCC replacement on DMI among treatments. In addition, digestibilities of DM, OM and CP were not different while aNDF linearly increased with an increasing level of GCC replacement. However, GCC replacement did not affect rumen fermentation such as ruminal pH, $NH_3$-N and VFA concentration; except C3 proportion which was the highest at 33% replacement while the lowest was at 100% replacement. All replacements of GCC resulted in similar protozoal and bacterial populations and microbial protein synthesis (MPS). Purine derivatives (PD) concentration in urine and PD to creatinine (PDC) index were varied with time of urination and among treatments at 0 to 8 and 8 to 16 h post feeding and higher values were shown among the GCC replacement groups. However at 16 to 24 h-post feeding, it was untraceable. In addition, creatinine concentration was similar among all treatments at every sampling time. Based on the above results, GCC can be used as an energy source for swamp buffalo fed with rice straw. Spot sampling of urine can be used for purine derivatives determination.
Darlis, N. Abdullah;Halim, R.A.;Jalaludin, S.;Ho, Y.W.
Asian-Australasian Journal of Animal Sciences
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v.13
no.7
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pp.922-928
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2000
The effects of animal species and supplements on rumen fluid characteristics, plasma urea-N (PUN) concentration, plasma urea-N pool size, urea-N degradation in the gut and urea-N net flux (urea-N synthesis rate) were studied in goats and sheep, with some minor differences detected. The animals were fed either chopped rice straw ad libitum+200 g soybean meal (SBM), or chopped rice straw ad libitum+190 g soybean meal+300 g sago meal (SBM+SM) for 14 days. The supplements were isonitrogenous (80 g crude protein/animal/d). [$^{14}C$]-urea was used as the marker for urea metabolism studies. Two animals from each species were fed either supplement in a cross-over design in two periods. The results showed that rumen pH was significantly (p<0.001) lower in animals fed SBM+SM than those fed SBM supplement. The ammonia concentrations of rumen fluid were significantly (p<0.01) higher in sheep (382.9 mg N/L) than goats (363.1 mg N/L) when fed SBM supplement but lower (282.5 mg N/L) than that of goats (311.0 mg N/L) when fed SBM+SM supplement. Total VFA concentrations were significantly (p<0.05) higher in animals fed SBM+SM supplement than those fed SBM supplement. Goats had significantly (p<0.01) higher molar proportions of acetate (79.1, 77.7%, respectively) than sheep (75.8, 74.0%, respectively) in both supplements. The molar proportion of acetate was significantly (p<0.05) higher, while that of butyrate lower in animals fed SBM supplement than those fed SBM+SM supplement. In animals fed SBM supplement, the molar proportion of propionate was significantly (p<0.01) higher in sheep (18.0%) than in goats (15.6%), but in animals fed SBM+SM, the molar proportion of butyrate was significantly (p<0.01) higher (9.6%) in sheep than in goats (7.2%). Plasma urea-N concentration, plasma urea-N pool size, urea-N degradation in the gut, urea-N net flux and the fraction of urea-C from the blood entering the rumen were not significantly different between goats and sheep fed either supplement. However, PUN concentration was significantly (p<0.05) lower in animals fed SBM+SM supplement (average of 13.8 mg N/100 ml) than in those fed SBM supplement (average of 16.5 mg N/100 ml). The urea net flux was significantly (p<0.05) higher in goats (average of 14.5 g N/d) than sheep (average of 12.9 g N/d), and animals fed SBM supplement showed higher (average of 14.9 g N/d) urea net flux than animals fed SBM+SM supplement (average of 12.9 g N/d). A significant (p<0.05) positive correlation was observed between urea-N net flux and urea-N degradation; urea-N net flux and pool size; urea-N net flux and urea excretion in the urine; and PUN and rumen ammonia in goats. While in sheep, significant (p<0.05) positive correlation was observed between urea-N net flux and urea excretion in the urine; and PUN and rumen ammonia.
Journal of the Korean Society of Food Science and Nutrition
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v.44
no.12
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pp.1905-1911
/
2015
Melanin is one of the most important factors affecting skin color. Melanogenesis is the bioprocess of melanin production by melanocytes in the skin and hair follicles and is mediated by several enzymes, such as tyrosinase, tyrosinase related protein (TRP)-1, and TRP-2. Convenient enzymatic transformation of the simple phenol pyrogallol with polyphenol oxidase originating from pear to an oxidative product, purpurogallin, was efficient. The structure of the pyrogallol oxidation product was identified on the basis of spectroscopic methods. The biotransformation product purpurogallin showed significant inhibitory effects against both melanin synthesis and tyrosinase activity in a dose-dependent manner in B16 melanoma cells. In addition, purpurogallin significantly attenuated melanin production by inhibiting TRP-1, and TRP-2 expression through modulation of their corresponding transcription factors, and microphthalamia- associated transcription factor in B16 cells. Consequently, purpurogallin derived from convenient enzymatic transformation of pyrogallol might be a beneficial material for reducing skin hyperpigmentation.
This study explores the interaction between the production of indole-3-acetic acid (IAA), a typical phytohormone auxin and the role of IAA biosynthetic pathways in each IAA producing rhizobacterial strain. The bacterial strains were isolated from rhizosphere of wild plants and identified as Acinetobacter guillouiae SW5, Bacillus thuringiensis SW17, Rhodococcus equi SW9, and Lysinibacillus fusiformis SW13. A. guillouiae SW5 exhibited the highest production of IAA using tryptophan-dependent pathways among the 4 strains. When indole-3-acetamide (IAM) was added, Rhodococcus equi SW9 showed the highest IAA production of $3824{\mu}g/mg$ protein using amidase activity. A. guillouiae SW5 also showed the highest production of IAA using two pathways with indole-3-acetonitrile (IAN), and its nitrile hydratase activity might be higher than nitrilase. B. thuringiensis SW17 showed the lowest IAA production, and most of IAA might be produced by the amidase activity, although the nitrilase activity was the highest among 4 strains. The roles of nitrile converting enzymes were relatively similar in IAA synthesis by Lysinibacillus fusiformis SW13. Tryptophan-independent pathway of IAA production was utilized by only A. guillouiae SW5.
Urea in the oral cavity is hydrolyzed mainly by bacterial ureases to ammonia, which in turn, raises pH of the oral environment, maintaining oral pH homeostasis, thereby inhibiting dental caries. Streptococcus salivarius has been shown to be a major contribution to oral ureolysis. Synthesis of urease by S. salivarius appears to be constitutive, but can be greatly enhanced in the acidic environment. It has been presumed that ureolytic activity of S. salivarius strains isolated from caries-active site is greater than that of strains from caries-free site. However, no in vivo study has supported the presumption. The present study was performed to observe the ureolytic activity of S. salivarius strains isolated from different environments in the same individual, finding out whether the ureolytic activity is related to dental caries. For the purpose, S. salivarius strains were isolated from caries-active site (>C2), a caries-free site of the tooth, and the dorsum of the tongue of each of 50 patients having decayed teeth. The strains isolated from the patients who harbored S. salivarius in more than two sites were selected and then their ureolytic activities were measured. In order to examine clonal diversity of the strains, their ureC genes were amplified by polymerase chain reaction (PCR) and then restricted with EcoRV, and the protein profiles of the strains were compared by SDS-PAGE. The results were as follows: 1. Of 50 patients, 13 patients harbored S. salivarius in more than two sites; a total of 61 S. salivarius strain were isolated from the patients and selected for the study. 2. Of 17 isolates from the caries-active site of 9 patients harboring S. salivarius in more than two sites including carious lesion, 10 (58.8%) showed a high ureolytic activity (> 200 ${\mu}mol/min/mg$). While, 19 out of 44 isolates (43.2%) from the caries-free site of the teeth and the dorsum of the tongues of 13 patients were the strains with a high ureolytic activity. 3. Of 9 patients harboring S. salivarius in more than two sites including caries-active site. 6 patients were found to have the strains in the caries-active site showing a lower ureolytic activity than the strains in the other sites. 4. Of 34 isolates with ureolytic activity higher than 40 ${\mu}mol/min/mg$, 32 isolates produced 0.54-Kbp PCR products regardless of the sites of bacterial collection. In contrast, of 27 isolates with ureolytic activity lower than 40${\mu}mol/min/mg$, 26 isolates yielded 1.3-Kbp PCR products or none regardless of the sites. 5. Different clonal types of S. salivarius with relatively higher and lower ureolytic activities were found in the same individuals and even in the same sites. 6. None of strains showing different ureolytic activity appeared to be the same clonal type. The overall results suggest that ureolytic activity of the isolates does not appear to be related to differences of the environments but related to their own genetic traits.
The activity of the liver cells of greenling, Agrammus agrammus were histologically investigated under photo-and electron microscopy, and studied by comparing seasonal changes of hepatosomatic index (HSI). The materials were monthly collected at the costal area of Tongbaeksom, Pusan, Korea, from September 1983 to August 1984. The annual variations of HSI of male were not distinct, but those of HSI in female began to increase in autumn, and reached the maximum in winter when the ovary was getting mature. During the period of yolk accumulation in the oocytes, the female liver and its hepatic cells were seen to large and nuclei and nucleoli were hypertrophic also. At this time the amounts of glycogen and lipid in the cells gradually decreased, while basophilic substance (RNA) increased. And well-developed granular endoplasmic reticula binding ribosomes were supposed to play the leading role in protein synthesis and deposition for vitellogenin in the cystoplasm. Just prior to spawning, glycogen and lipid droplets were decreased, but basophilic substances(RNA) were found in a high concentration especially at the peripheral region of the liver cells of females. In the liver cells of males, were hardly altered by gonadal maturation, basophilic substances gradually increased, glycogen particles and lipid droplets were still observed in large quantities. After spawning, basophilic subtances decreased in the liver cells of female and male.
Journal of the Korean Society for Marine Environment & Energy
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v.19
no.2
/
pp.144-150
/
2016
In southwestern East Sea, we investigated the spatio-temporal distribution characteristics of particulate organic carbon (POC) and nitrogen (PON) in September 2011 (summer), January (winter) and May 2012 (spring). Although cold waters known as the origin upwelling in the surface layer of September were not observed, this periods showed high primary productivity because of high concentrations of chlorophyll, low percentage of non-autotrophic particulate fraction among POC calculated by POC/Chl-a ratio (27%) and low POC/PON ratio (6.2), which means active amino acid and protein synthesis, However, May, 2012 showed low primary productivity because of high percentage of non-autotrophic particulate fractions among POC (66%) and high POC/PON ratio (8.1), Although spring bloom and high primary productivity has been reported in the East Sea, high percentage of non-autotrophic particulate fractions in POC, observed in the East sea during the post 2012 spring, is suggested to be due to the increase of phaeo-pigment during post spring bloom. Thus, composition of particulate organic matter may have sensitively changed by marine environmental factors in spite of same season.
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