Eighty-five different cultivars of Brassica rapa, B. juncea, B. nap us, B. carinata, B. oleracea and hexaploid Brassica collected from Bangladesh, Japan, China and Denmark were analyzed by SDS-PAGE for seed and leaf protein variations, using esterase, acid phosphatase and peroxidase isozyme analysis. Ten polymorphic bands were identified from seed protein however no identifiable polymorphic band was found in the leaf protein. Polymorphic markers clearly distinguished the different Brassica species as well as yellow sarson (YS) and brown seeded (BS) cultivars of B. rapa. The $F_1$ cross between YS and brown seeded cultivars showed the existance of all poly-morphic bands of the respective parents. The Bangla-deshi and Japanese cultivars of B. rapa differed in the amount of seed protein. In the case of isozyme analysis, esterase showed the highest number of polymorphic bands (13) followed by acid phosphatase (9) and peroxidase (5). These polymorphic markers were very effec-tive for classification of all the species studied in this experiment. In parentage tests using isozymes, the hybridity of intra-and-interspecific crosses of almost all the seedlings could be identified from their respective cross combinations. Esterase polymorphism showed a clear differentiation between YS and BS types of B. rapa. In addition, two esterase polymorphic markers were iden ified to differentiate some cultivars of B. juncea. Segregation patterns in these two esterase bands showed a simple Mendelian monohybrid ratio of 3:1 in $F_2$, 1:1 in test cross and 1:0 in back cross progenies. No polymorphic band was identified to distinguish different cultivars of the same species by acid phosphatase or peroxidase. Polymerase Chain Reaction (PCR) was carried out with seed coat color specific marker of B. juncea. The yellow seeded cultivars produced a strong band at 0.5 kb and weak band 1.2 kb. In the addition of these two specific bands, Japanese yellow-seeded cultivars expressed two more weak bands at 1.0 kb and 1.1 kb. Where the brown seeded cultivars generated a single strong band at 1.1 kb. In segregating population, the yellow seed coat color marker segregated at a ratio 15 (brown) : 1 (yellow), indicating the digenic inheritance pattern of the trait.
The effect of orthodontic force on the collagenase and phosphatase activities of the adjacent alveolar bone was evaluated. Maxillary canines of male cats were treated orthodontically with closed coil spring so as to exert about 80g force. Sixteen cats were equally divided into one control group and seven experimental groups (12 hrs, 24 hrs, 36 hrs, 2 days, 3 days, 5 days and 7 days after orthodontic treatment). After sacrificing all animals on experimental intervals, collagenase, acid phosphatase (ACP) and alkaline phosphatase (ALP) activities were determined in the pressure and tension sides of alveolar bones. ACP activities increased in both the pressure and tension sides, but significantly increased in the pressure side continuously. ALP activities increased in the tension side at early stage (1-2 days after treatment), but changed small amount in the pressure side. Collagenase activities increased in the pressure side, especially at late stage (5-7 days after treatment). These results suggest that (1) orthodontic fore force increases the ACP, ALP and collagenase activities generally and (2) activities of ACP and collagenase increase in the pressure side, but that of ALP in the tension side and (3) activities of ACP and ALP increase at early stage, but that of collagenase at late stage after orthodontic treatment. Therefore it is shown that there are time differences in the formation and destruction of organic and inorganic components in the bone metabolism of alveolus with application of the orthodontic forces.
Panax ginseng occupies an important role in the folk medicine of China, Korea and Japan. The present study was undertaken to determine the radioprotective efficacy of ginseng root extract in the liver of Swiss albino mice. The animals were divided into 4 groups. Group I-Only vehicle was administered. Group II-The animals received 10 mg/kg body weight ginseng root extract i.p. for 4 consecutive days. Group III-Animals were irradiated with 8Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 ems. Group IV-Animals were given by ginseng root extract (10 mg/kg body weight) continuously for 4 days and on 4th day they were irradiated with 8 Gy gamma radiation after 30 min. The animals from above groups were autopsied on 1,3,7,14 and 30 days. Biochemical estimations of phosphatases (acid & alkaline), LDH (lactate dehydrogenase), LPO (lipid peroxidation) and GSH (reduced glutathione) in liver and SGOT (serum glutamate oxaloacetate transaminase), SGPT (serum glutamate pyruvate transaminase) and alkaline phosphatase in serum were done. In ginseng treated group acid phosphatase (ACP), alkaline phosphatase (ALP), LPO and LDH in liver and SGOT, SGPT and alkaline phosphatase in serum did not show any significant alteration. However, a significant increase in GSH content in liver was recorded. In irradiated group there was a significant increase in ACP, ALP and LPO content in liver and SGOT & SGPT in serum was noted. Whereas, a significant decrease was recorded in GSH and LDH activity in liver and alkaline phosphatase activity in serum. Pretreatment of ginseng with radiation significantly alters the biochemical parameters in liver and serum. A significant decline in ACP, ALP activity and LPO content in liver and SGOT and SGPT activity in serum was observed. However, a significant increase in GSH content and LDH activity in liver and ALP activity in serum was estimated. The present study suggests that pretreatment of ginseng before irradiation significantly protects the liver and maintains the enzyme activity.
Butyrate is one of the short-chain fatty acids that are present in the colon of mammals in millimolar concentration as a result of microbial anaerobic fermentation of dietary fiber, undigested starch, and proteins. In this study, sodium butyrate was examined in HT29 cell, human colonic cancer cell line, on cell viability, alkaline phosphatase activity, PLC-${\gamma}$1 expression and complex sphingolipid biosynthesis. Treatment with butyrate showed that the decrease of cell adhesion and viability was time-dependent. Sodium butyrate also induced to increase the activity of alkaline phosphatase which is a differentiation marker enzyme and decrease the expression of PLC-${\gamma}$1. Biosynthesis of sphingomyelin and galactosylceramide by butyrate treatment were decreased so fast but ceramide was increased 680dpm/mg protein% more than untreated group on first day and then decreased fast. In addition, acid ceramidase and neutral ceramidase activity were inhibited early stage by sodium butyrate. These results suggest that sodium butyrate causes cell differentiation or cell growth arrest of HT29 cell accompanied by early increase of ceramide content and alkaline phosphatase activity and decrease of galactosylceramide content and PLC-r1 expression.
Jang, Sei-Heon;Hwang, Si Ae;Kim, Mijin;Yun, Sung-Hae;Kim, Moon-Sook;Karnik, Sadashiva S.;Lee, ChangWoo
Molecules and Cells
/
v.28
no.1
/
pp.25-30
/
2009
${\beta}$-Arrestins turn off G protein-mediated signals and initiate distinct G protein-independent signaling pathways. We previously demonstrated that angiotensin $AT_1$ receptorbound ${\beta}$-arrestin 1 is cleaved after $Phe^{388}$ upon angiotensin II stimulation. The mechanism and signaling pathway of angiotensin II-induced ${\beta}$-arrestin cleavage remain largely unknown. Here, we show that protein Tyr phosphatase activity is involved in the regulation of ${\beta}$-arrestin 1 cleavage. Tagging of green fluorescent protein (GFP) either to the N-terminus or C-terminus of ${\beta}$-arrestin 1 induced conformational changes and the cleavage of ${\beta}$-arrestin 1 without angiotensin $AT_1$ receptor activation. Orthovanadate and molybdate, inhibitors of protein Tyr phosphatase, attenuated the cleavage of C-terminal GFP-tagged ${\beta}$-arrestin 1 in vitro. The inhibitory effects of okadaic acid and pyrophosphate, which are inhibitors of protein Ser/Thr phosphatase, were less than those of protein Tyr phosphatase inhibitors. Cell-permeable pervanadate inhibited angiotensin II-induced cleavage of ${\beta}$-arrestin 1 in COS-1 cells. Our findings suggest that Tyr phosphorylation signaling is involved in the regulation of angiotensin II-induced ${\beta}$-arrestin cleavage.
The present investigation has been undertaken to understand the mechanism of implantation process, by demonstrating the role of ovarian steroids in connection with phosphatase activity in the differentiation of uterine endometrium for implantation. The results obtained are as followings: The differentiation of the uterine endometrial tissue was closely influenced by the ovarian steroid hormones; at first, 17${\beta}$-estradiol initiated the differentiation of the uterine luminal and glandular epithelial cells, and then progesterone induced differentiation of stromal cells, and thereby two steroids maintain decidualization of the uterine tissues. We observed that the phosphatase activities seem to be dependent upon the ovarian steroids; that is the activity showed higher level in progesterone treated group than in estradiol treated one, and the highest activity was found in the group treated with both estradiol and progesterone. Acid phosphatase showed the highest activity whereas alkaline phosphatase showed the lowest in the rat uterine endometrium during early pregnancy.
Megagametophyte tissues of Pinus densiflora were subjected to study the inheritance of acid phosphatase (ACP), alcohol dehydrogenase (ADH) and catalase (CAT) isozymes by starch gel zone-electrophoresis. At least three or four zones were segregated for ACP isozyme. However, as one isozyme of ACP-A zone was separated clearly, only that isozyme was analysed. Five isozyme phenotypes (A1-A5), observed in ACP-A zone, were segregated to a simple Mendelian ratio, suggesting that these are controlled by five codominant alleles existed at ACP-A locus. Two zones of activity were segregated in the gels after staining for ADH, the more anodal zone (ADH-A) of the two was invariant in our materials. Three isozyme phenotypes (B1-B3) were observed in ADH-B zone and these variants showed a 1:1 segregation pattern, suggesting that each variant is controlled by three codominant alleles at ADH-B locus. A total of five isozyme phenotypes, composed of multiple bands, were observed in CAT isozyme. The segregation of these phenotypes in heterozygous trees did not show any significant deviation from a 1:1 segregation. Therefore, the genetic control of CAT isozyme in Pinus densiflora seeds seems to be based on a single locus (CAT-A) with Five codominant alleles ($A_1-A_5$).
This study was performed to examine the effect of bisphosphonate, an inhibitor of bone resorption, on the formation of osteoclast and bone resorption during experimental tooth movement. Whether bisphosphonate has a cytotoxicity in high dose was also examined. Eighty-seven male Sprague-Dawley rats, weighing 260-350g, were classified into normal (no appliance + $0.9\%$ NaCl), control (appliance + $0.9\%$ NaCl) and four bisphosphonate-treated (appliance + 0.8, 4, 20 or 100mg/kg) groups. The maxillary left first molar was moved mesially with the tipping movement using 50-70g of force. Bisphosphonate(etidronate disodium) was injected intraperitoneally with a dose of 0.8, 4, 20, or 100 mg/kg simultaneously with the application or the orthodontic force. They were killed at day 1, 3, or 7 after the application or the orthodontic force. The activities of serum acid phosphatase and lactate dehydrogenase (LDH) were assayed, and osteoclasts and the degree of bone resorption were examined histologically. The results obtained were as follows: 1. Acid phosphatase activities were significantly higher in the appliance groups, both control and bisphosphonate-treated (4, 20, and 100 mg/kg) groups, at days 1 and 3 than these in normal. At day 1, bisphosphonate-treated(4, 20mg/kg) groups showed even higher acid phosphatase than control. However, at day 7, no significant difference was noted between the control and bisphosphonate-treated groups. 2. LDH activities in the 4, 20mg/kg bisphosphonate-treated groups were increased during the experimental Periods examined, but there were no significant differences in the 0.8, 100mg/kg bisphosphonate-treated groups. 3. There was no bone resolution at day 1, but severe bone resorption was observed at days 3 and 7 in the control. Bone resorption was reduced by bisphosphonate-treatment at day 3. Bone resolution observed at day 7 was similar between the control and bisphosphonate-treated groups. 4. Few osteoclasts were observed at the alveolar bone in the control and bisphosphonate-treated groups at day 1. At day 3, numerous osteoclasts were shown in the control, the degree of which was reduced in bisphosphonate-treated groups. These results suggest that the inhibition of the osteoclast formation was not the mechanism of bone resorption by the bisphosphonate-treatment during experimenal tooth movement. There was no distinct cytotoxicity with a high dose of bisphosphonate. And the drug should be administrated repeatedly to maintain the inhibitory effect of bone resolution.
Objective: The study was conducted to investigate the dephosphorylation of Pseudomonas aeruginosa flagellin (PA FLA) by sweet potato purple acid phosphatase (PAP) and the effect of the enzyme on the flagellin-mediated inflammatory response in the A549 lung epithelial cell line. Methods: The activity of sweet potato PAP on PA FLA was assayed at different pH (4, 5.5, 7, and 7.5) and temperature (25℃, 37℃, and 55℃) conditions. The release of interleukin-8 (IL-8) and the activation of nuclear factor kappa- light-chain-enhancer of activated B cells (NF-κB) in A549 cells exposed to PA FLA treated with or without sweet potato PAP was measured using IL-8 and NF-κB ELISA kits, respectively. The activation of toll-like receptor 5 (TLR5) in TLR5-overexpressing HEK-293 cells exposed to PA FLA treated with or without sweet potato PAP was determined by the secreted alkaline phosphatase-based assay. Results: The dephosphorylation of PA FLA by sweet potato PAP was favorable at pH 4 and 5.5 and highest at 55℃. PA-FLA treated with the enzyme decreased IL-8 release from A549 cells to about 3.5-fold compared to intact PA FLA at 1,000 ng/mL of substrate. Moreover, PA-FLA dephosphorylated by the enzyme repressed the activation of NF-κB in the cells compared to intact PA FLA. The activation of TLR5 by PA-FLA was highest in TLR-overexpressing HEK293 cells at a substrate concentration of 5,000 ng/mL, whereas PA FLA treated with the enzyme strongly repressed the activation of TLR5. Conclusion: Sweet potato PAP has the potential to be a new alternative agent against the increased antibiotic resistance of P. aeruginosa and may be a new conceptual feed additive to control unwanted inflammatory responses caused by bacterial infections in animal husbandry.
Dry powder and different extracts of Agave lophantha were tested against Biomphalaria alexandrina. The results showed that the butanol extract has high molluscicidal activity. The activity of the dry powder has been found to be stable under the effect of some simulated field conditions. Also the toxicological effect of the plant on mice was tested through determination of certain parameters such as total protein, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and acid phosphatase enzymes as well as histopathological study on liver and kidney.
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