Human prostatic acid phosphatase (PAP), with comprehensive homology to glandular kallikrein, are representative serum biomarkers of prostate cancer. Dendritic cell (DC), which is the potent antigen-presenting cells(APC) in the immune system, can induce strong T cell responses against viruses, microbial pathogens, and tumors. Therefore, the immunization using DC loaded with tumor-associated antigens is a powerful method for inducing anti-tumor immunity. The CTP (Cytoplasmic Transduction Peptide) technology developed by Creagene which can transport attached bio-polymers like nucleic acids or proteins into the cell with high permeation efficiency. As the active forms of PAP can mediate apoptotic processing, we used multimer forms of PAP as an inactive form for antigen pulsing of DCs. In this study, multimeric forms of CTP-rhPAP was obtained according to the advanced purification process and subsequently confirmed by gel filtration chromatography, western blot and Dynamic Light Scattering. Therefore, CTP-conjugated PA multimers transduced into the cytoplasm were efficiently presented on the cell surface without any harm effect on cells via MHC class I molecules and result in induction of a large number of effector cell.
Purpose: Ursodeoxycholic acid (UDCA) is known to decrease hepatic injury by promoting the biliary secretion of retained toxic endogenous bile acids in hepatobiliary diseases complicated by total parenteral nutrition (TPN). However, most studies have focused on treatment for complications after TPN. We investigated the preventive role of early administration of UDCA in TPN-induced hepatobiliary complications by a randomized, double-blind, placebo-controlled trial. Methods: Between May 2000 and May 2002, thirteen patients, who were given TPN more than 10 days in the hospital, were assigned randomly to two groups. One was the case group (7 patients) who were given UDCA simultaneously with TPN regimen, and the other, the control group (6 patients) who were given placebo. Their age ranged from 1 day to 13 years. They were affected with diseases impossible for enteral nutrition, such as prematurity, cerebral palsy, chronic diarrhea, anorexia nervosa, pancreatitis, and cyclic vomiting. The duration of TPN ranged from 10 to 70 days. Hematologic parameters including liver function test were measured at regular intervals, and the duration, composition, administration rate, total calorie of TPN were recorded. The serum levels of total bilirubin, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase were compared between groups after cessation of the study. Results: The autoregressive coefficient of the control group was 0.4419 (p=0.0651) in bilirubin, -0.0431 (p=0.7923) in AST, 0.2398 (p=0.2416) in ALT, and 0.2459 (p=0.1922) in alkaline phosphatase by mixed procedure model when the parameters were referred to the case group. Conclusion: The serum level of total bilirubin did not increase in comparison with that of the control group, but statistically insignificant, when both TPN and UDCA were administered simultaneously from the beginning.
Ki yong Kim;Dae yuong Son;Yong Gu Park;Won Il Jung;Jin Ki Jo
Journal of The Korean Society of Grassland and Forage Science
/
v.13
no.3
/
pp.177-183
/
1993
This study was conducted to obtain the transformed tobacco plants with S. cerevisiae Acid phosphatase gene(PH05) using Agrobacterium tumefaciens and th confirm plant transformation and gene expression. the results obtained were summarized as follows: APase activity of Saccharomyces cereviase NA 87-11A was remarkably showed up as deep red color when assayed by Tohe and Oshima(1974). PH05 fragment, Apase gene, was obtained from pVC727G and the graphically estimated size was about 1.5kb by agarose gel electrophoresis. The sequencing results of 5'end and 3'end of PH05 using dideoxy chain termination method were coinsided with the full length nucleotide already. pBKJ I vector was constructed by isolation of PH05 fragment from pVC727-1 and pBKSI-1 digesred with Sma I and Xba I. Isolated plasmid from transformed A. tumefaciens with constructed pBKJ I when it was electrophoresed with agarose gel. The dosc of tobacco leaf was cocultivated 재소 transformed Agronacterium tumefaciens. Transformed shoots were selected on kanamtcin-containing MS-n/B medium and they were regenerated. The transgenic tobacco plants were elucidated by isolation of genomic DNA and genomic southern hybridization using ${\alpha}-^{32}P$ labelled PH05 fragments. The PH05 in transformed tobacco plants was expressed in leaf, stem and root, and its APase activity was estimated as deep red color by Tohe method.
Gold compounds depress phagocytic cell responses, including chemotaxis, and respiratory burst. However, the effects of gold compounds on the function of phagocytic cells are variable according to the preparation of medicine. In this study, effect of tetrachloroauric acid on activated neutrophil responses, including respiratory burst, lysosomal enzyme release and change of intracellular $Ca^{2+}$ level and on the synthesis of interleukin-8 and granulocyte-macrophage colony stimulating factor by macrophages was studied. This study further examines how gold compounds affect the activation processes. The respiratory burst stimulated by complement C5a, degraded IgG and PMA in neutrophils was inhibited by tetrachloroauric acid. In contrast to C5a and degraded IgG, PMA-stimulated superoxide production was weakly inhibited by tetrachloroauric acid. Staurosporine, genistein, EGTA and verapamil inhibited superoxide and $H_2O_2$ production caused by C5a and degraded IgG. PMA-stimulated superoxide production was inhibited by staurosporine but was not affected by genistein. Tetrachloroauric acid, genistein, EGTA and verapamil inhibited the release of acid phosphatase and myeloperoxidase, while the effect of staurosporine was not detected. The synthesis of interleukin-8 and granulocyte-macrophage colony stimulating factor by $interleukin-1{\beta}$ in macrophages was inhibited by tetrachloroauric acid. Preincubation with tetrachloroauric acid, genistein, EGTA and verapamil, the elevation of [$Ca^{2+}_i$] evoked by C5a was inhibited. Store-regulated $Ca^{2+}$ entry in thapsigargin-pretreated neutrophils was decreased by the addition of tetrachloroauric acid and genistein. The effect of staurosporine on intracellular $Ca^{2+}$ mobilization was not observed. In conclusion, tetrachloroauric acid may suppress neutrophil responses through its inhibitory action on elevation of intracellular $Ca^{2+}$ level and protein kinase C. It might exhibit an inhibitory effect on the action of protein tyrosine kinase. Tetrachloroauric acid depresses cytokine production by macrophages.
Park, Kyoung Soo;Shin, Myung Seok;Chang, Mea Young
Clinical and Experimental Pediatrics
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v.49
no.8
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pp.851-856
/
2006
Purpose : The purpose of this study is to find out the diagnostic significance of serum bile acid on total parenteral nutrition induced cholestasis in premature infants. Methods : Infants without cholestasis were classified into postnatal days and each change of serum bile acid was measured and analyzed. Also, the serum direct bilirubin, serum bile acid, ${\gamma}$-glutamic acid transferase, and alkaline phosphatase of premature infants with total parenteral nutrition induced cholestasis were measured for comparison and analysis of their correlation. Results : Changes of serum bile acid analysis after birth showed no significant difference between boys and girls, between premature infants and term infants without cholestasis. Serum bile acid levels are constant after two weeks after birth in neonates without cholestasis. In premature infants with total parenteral nutrition induced cholestasis, the increase of serum direct bilirubin over 2 mg/dL was $34.9{\pm}18.3$ days after birth, and the increase of serum bile acid was $28.1{\pm}18.3$ days. Its increase was about 1 week faster than serum direct bilirubin, however, there was no statistical significance(P=0.114). Comparing analysis of serum bile acid, ${\gamma}$-glutamic acid transferase, and alkaline phosphatase, serum bile acid showed the highest correlation to serum direct bilirubin(r=0.487, P=0.000). Conclusion : Serum bile acid is an important parameter of total parenteral nutrition induced cholestasis in premature infants and will be useful for early diagnosis and treatment.
The study was intended to know any relations between the rice tolerance to bensulfuron and varietal speciation in seed protein composition or any enzymatical allelies with or without chemical treatment. Rice varieties used were UCP-28, Chinsurah Boro II, Fukunohama, Fadehpur-2, IR 14252-13-2-2-5 as the tolerant group, and HP 93(3) FA, HP94(9) FA, Padilabou Alumbis, KH-17854, and IR 1846-2841-1 as the susceptible, respectively. Electrophoretic methods used were SDS-PAGE for seed protein, 7% PAGE for isozymes (acid phosphatase, peroxidase, malate dehydrogenase, and esterase from rice seedling) and variation in isoenzyme profiles (malate dehydrogenase, peroxidase, and esterase) as affected by different concentrations of bensulfuron(0, $10^{-6}$, $10^{-5}$ and $3{\times}10^{-5}M$) was also studied. The results are summarized as follows. -Among 16 bands separated in seed proteins, two different rice groups selected in terms of tolerance to bensulfuron were clustered in dissimilarity, which was based on relatively larger area in whole peaks and higher activities in N, O, P bands for the tolerant group. -Among isozymes obtained from rice seedlings without chemical treatments, the following specificities were obtained. The tolerant varieties had the relatively higher activity in D band out of 4 peroxidase bands. Malate dehydrogenase was separated into 3 bands and only tolerant varieties had A band and higher activities in Band C bands. Esterase was separated into 3-4 bands with higher activities in A and B bands for tolerant varieties. There were one major band accompanied by 2-3 minor bands for acid phosphatase in which only tolerant varieties had the B band. -The effect of Bensulfuron concentration on the isozyme activities showed that the activity of C band in peroxidase was not present in tolerant varieties which was contrary to the increased activities in susceptible varieties. However, D band was gradually disappeared only in susceptible varieties as the concentration of bensulfuron was increased. For malate dehydrogenase in the susceptible varieties, major bands D, E and F kept consistantly higher activities while minor bands A, B and C disappeared sensitively. Among 5 bands of esterase separated, D band was present only in the tolerant varieties while E band only in the susceptible. The activities in A, C, E bands were sharply decreased in the susceptible varieties as the concentration of bensulfuron was increased.
Five different secretion vectors were constructed by varying the signal sequences and .alpha.-antitrypsin (.alpha.1-AT) a numan secretory protein, was produced from yeast cells. The signal sequences used are those of acid phosphatase (PH05) and .alpha.-factor (M f.alphal1) of Saccharomyces cerevisiae, glucoamylase (STA1) of Saccharomyces diastaticus, and human .alpha.1-AT. Four vectors directed the efficient secretion of .alpha.1-AT ito the culture media. The secretion vector carrying the glucoamylase signal sequence (pGAT11) showed the highest efficiency of secretion. About 70% of .alpha.1-AT produce dwere secreted into the media. The endo H treatment of partially purified .alpha.1-AT indicates that the secreted .alpha.1-AT appeared to be glycosylated. This glycosylation pattern was altered when amino acid substitution mutations were introduced at the three glycosylation sites of .alpha.-AT.
The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of periodontal ligament cells. Primary human periodontal ligament cells were cultured in dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells of 4th to 7th passage were inoculated in the multiwell plates coated with chitosan in concentration of 0.22, 0.2, and $2mg/m{\ell}$. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized modules was evaluated after 21 days of culture. The results were as follows : 1. The morphology of periodontal ligament cells on the chitosan coating was round or spheric. Round cells were aggregated after 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule-like appearance after 24 hours of culture and not achieved confluency at 7 days. 2. During early period of culture, the attachment of periodontal ligament cells were inhibited by chitosan coating. Inhibition of cell attachment tended to increase with the concentration of chitosan. 3. At the chitosan concentration of 0.02 and $0.2mg/m{\ell}$, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of $2mg/m{\ell}$, the proliferation of periodontal ligament cells was inhibitied(p<0.01). 4. Alkaline phosphatase activity of periodontal ligament cells was increased in chitosan coated group, especially at the concentration of $0.02mg/m{\ell}$after 4 days of culture.5. Periodontal ligament cells produced mineralized nodules on chitosan coated wells without the addition of mineralized nodule forming materials (ascorbic acid, ${\beta}-glycerophosphat$, dexamethasone). With the addition of mineralized nodule forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of $0.02mg/m{\ell}$, compared to the control. In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration($0.02mg/m{\ell}$), chitosan could increase alkaline phosphatase activity and stimulate the formation of mineralized nodule by periodontal ligament cells. These results suggest that chitosan can be used as an adjunct for bone graft material, and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.
Changes of contents of reducing sugar and sucrose and activities of sucrose-phosphate synthase, sucrose synthease and invertase from the leaves of barley (Hordeum vulgare L. cv. Chalssal) seedlings grown at $4^{\circ}C$ were investigated, and the property of acid invertase were also examined. In the seedlings grown at $4^{\circ}C$ for 3 days, the contents of reducing sugar and sucrose were increased to 1.3 and 2.4 times, respectively. Activity of acid invertase was decreased markedly by cold treatment while the activities of sucrosephosphate synthase, sucrose synthase, and alkaline invertase were not changed. In acid phosphatase purified partially by ammonium sulfate fractionation and DEAE-Sephacel column chromatography, the $K_m$ value for sucrose was 9.5 mM and the optimum pH and temperature was 5.5 and $35^{\circ}C$ respectively. This enzyme was supposed to be ${\beta}-fructosidase$ by studies on the substrate specificity and the molecular weight was estimated to be 63 Kd by Sephadex G-200 gel chromatography.graphy.
Kim, Seongjun;Han, Seung Hyun;Li, Guanlin;Yoon, Tae Kyung;Lee, Sang-Tae;Kim, Choonsig;Son, Yowhan
Journal of Ecology and Environment
/
v.40
no.1
/
pp.5-11
/
2016
Background: As the decomposition of lignocellulosic compounds is a rate-limiting stage in the nutrient mineralization from organic matters, elucidation of the changes in soil enzyme activity can provide insight into the nutrient dynamics and ecosystem functioning. The current study aimed to assess the effect of thinning intensities on soil conditions. Un-thinned control, 20 % thinning, and 30 % thinning treatments were applied to a Larix kaempferi forest, and total carbon and nitrogen, total carbon to total nitrogen ratio, extractable nutrients (inorganic nitrogen, phosphorus, calcium, magnesium, potassium), and enzyme activities (acid phosphatase, ${\beta}$-glucosidase, ${\beta}$-xylosidase, ${\beta}$-glucosaminidase) were investigated. Results: Total carbon and nitrogen concentrations were significantly increased in the 30 % thinning treatment, whereas both the 20 and 30 % thinning treatments did not change total carbon to total nitrogen ratio. Inorganic nitrogen and extractable calcium and magnesium concentrations were significantly increased in the 20 % thinning treatment; however, no significant changes were found for extractable phosphorus and potassium concentrations either in the 20 or the 30 % thinning treatment. However, the applied thinning intensities had no significant influences on acid phosphatase, ${\beta}$-glucosidase, ${\beta}$-xylosidase, and ${\beta}$-glucosaminidase activities. Conclusions: These results indicated that thinning can elevate soil organic matter quantity and nutrient availability, and different thinning intensities may affect extractable soil nutrients inconsistently. The results also demonstrated that such inconsistent patterns in extractable nutrient concentrations after thinning might not be fully explained by the shifts in the enzyme-mediated nutrient mineralization.
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