The aim of this study was to investigate the absorption properties of ketoprofen. The in-situ perfusion model has advantages over in vitro models as it provides intact lymphatic and blood flow circulation. The absorption properties of six different concentrations of ketoprofen have been studied in single pass in-situ rat intestine model. $^{14}C-PEG$ 4000 was used as a permeability marker and the possibility of an energy dependent contribution to ketoprofen absorption was also Investigated using the metabolic inhibitor sodium azide. Three different concentrations of sodium azide were studied to examine its effect on absorption of ketoprofen from the rat intestine. The findings of this study suggest that mono-carboxylic type drugs like ketoprofen cause permeability changes in the intestine. This is shown by the increase in absorption of $^{14}C-PEG$ 4000 as the concentration of ketoprofen is increased. However, the trend for ketoprofen permeability is to decrease over the concentration ranges. It was observed that the Papp values for ketoprofen with sodium azide shows a trend towards reduction in the amount of ketoprofen absorbed from the rat intestine which was significantly different (p<0.05) from that of ketoprofen with sodium azide 3.0mM. This indicates that sodium azide has an affect on the absorption of ketoprofen. The pH of all the perfusion solutions was altered to ${\sim}pH\;6.7$ by the buffering capacity of the small intestine secretions. The results suggest that mechanisms other than passive diffusion may be involved in ketoprofen absorption. This would be consistent with the involvement of active transport or saturatable processes in the absorption of drugs containing monocarboxylic acid group, as has been previously suggested from in vitro data.
Cadmium was transported through the special carrier system into the cells of cotyledons, hypocotyls, and roots spring radish young plants. The transport of cadmium was inhibited by metabolic inhibitor, like DNP. The $K_m$ values for cadmium were 0.7 ppm for cotyledons, 1.72 ppm for hypocotyls, and 0.3 ppm for roots, and $V_{max}$ for cadmium was $40ppm{\cdot}h^{-1}{\cdot}g{\cdot}fresh\;weight^{-1}$ for cotyledons, $313ppm{\cdot}h^{-1}{\cdot}g{\cdot}fresh\;weight^{-1}$ for hypocotyls, and $606ppm{\cdot}h^{-1}{\cdot}g{\cdot}fresh\;weight^{-1}$ for roots. Cadmium cannot prove to be inducer for proline accumulation. Therefore, proline accumulation cannot be used as a marker to test the level of heavy metal pollution.
Based on the potential beneficial effects of growth hormone releasing peptide (GHRP)-6 on muscle functions, a newly synthesized GHRP-6-biotin conjugate was tested on cultured myoblast cells. Increased expression of myogenic marker proteins was observed in GHRP-6-biotin conjugate-treated cells. Additionally, increased expression levels of insulin-like growth factor-1 and collagen type I were observed. Furthermore, GHRP-6-biotin conjugate-treated cells showed increased metabolic activity, as indicated by increased concentrations of energy metabolites, such as ATP and lactate, and increased enzymatic activity of lactate dehydrogenase and creatine kinase. Finally, binding protein analysis suggested few candidate proteins, including desmin, actin, and zinc finger protein 691 as potential targets for GHRP6-biotin conjugate action. These results suggest that the newly synthesized GHRP-6-biotin conjugate has myogenic stimulating activity through, at least in part, by stimulating collagen type I synthesis and several key proteins. Practical applications of the GHRP-6-biotin conjugate could include improving muscle condition. [BMB Reports 2015; 48(9): 501-506]
Seo, Hyun-Ju;Cho, Young-Eun;Kim, Tae-Wan;Shin, Hong-In;Kwun, In-Sook
Nutrition Research and Practice
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v.4
no.5
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pp.356-361
/
2010
Zinc is an essential trace element required for bone formation, however not much has been clarified yet for its role in osteoblast. We hypothesized that zinc would increase osteogenetic function in osteoblasts. To test this, we investigated whether zinc treatment enhances bone formation by stimulating osteoblast proliferation, bone marker protein alkaline phosphatase activity and collagen synthesis in osteoblastic MC3T3-E1 cells. MC3T3-E1 cells were cultured and treated with various concentrations of zinc (0, 1, 3, 15, 25 uM) along with a normal osteogenic medium (OSM) as control for 1, 5, 10 days. As measured by MTT assay for mitochondrial metabolic activity, cell proliferation was stimulated even at low zinc treatment (1-3 ${\mu}M$) compared to OSM, and it was stimulated in a zinc concentration-dependent manner during 5 and 10 days, with the most pronounced effect at 15 and 25 uM Zn. Cellular (synthesized) alkaline phosphatase (ALP) activity was increased in a zinc concentration-dependent manner, so did medium (secreted) ALP activity. Cellular collagen concentration was increased by zinc as time went by, therefore with the maximum zinc stimulatory effect in 10 days, and medium collagen concentration showed the same pattern even on 1 and 5 day. This zinc stimulatory effect of collagen synthesis was observed in cell matrix collagen staining. The study results imply that zinc can increase osteogenic effect by stimulating cell proliferation, ALP activity and collagen synthesis in osteoblastic cells.
Park, Myeong-Soo;Kang, Yoon-Hee;Cho, Sang-Hee;Seo, Jeong-Min;Ji, Geun-Eog
Proceedings of the Korean Society for Applied Microbiology Conference
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2001.06a
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pp.155-157
/
2001
Bifidobacterium spp. is nonpathogenic, gram-positive and anaerobic bacteria, which inhabit the intestinal tract of humans and animals. In breast-fed infants, bifidobacteria comprise morethan 90% of the gut bacterial population. Bifidobacteria spp. are used in commericial fermented dairy products and have been suggested to exert health promoting effects on the host by maintaining intestinal microflora balances, improving lactose tolerance, reducing serum cholesterol levels, increasing synthesis of vitamins, and aiding the immune enchancement and anticarcinogenic activity for the host. These beneficial effects of Bifidobacterium are strain-related. Therefore continued efforts to improve strain characteristics are warranted. in these respect, development of vector system for Bifidobacterium is very important not only for the strain improvement but also because Bifidobacterium is most promising in serving as a delivery system for the useful gene products, such as vaccine or anticarcinogenic polypeptides, into human intestinal tract. For developing vector system, we have characterized several bifidobacterial plasmids at genetic level and developed several shuttle vectors between E. coli and Bifidobacterium using them. Also, we have cloned and sequenced several metabolic genes and food grade selection marker. Also we have obtained bifidobacterial surface protein, which will be used as the mediator for surface display of foreign genes. Recently we have succeeded in expressing amylase and GFP in Bifidobacterium using our own expression vector system. Now we are in a very exciting stage for the molecular breeding and safe delivery system using probiotic Bifidobacterium strains.
Chlorella is a eukaryotic microalgae which shares metabolic pathways with higher plants. These charac-teristics make chlorella a potential candidate for eukaryotic overexpression systems. Recently, a foreign flounder growth hormone gene was stably introduced and expressed in transformed Chlorella ellipsoidea by using a modified plant transformation vector that contains cauliflower mosaic virus (CaMV) 35S pro-moter and the phleomycin resistant Sh ble gene as a selection marker. In this study, this same vector was modified by incorporating a promoter and a 3' UTR region of the 33kDa peptide gene from a chlorella virus that was isolated in our laboratory. The 33kDa gene promoter was used to replace the 35S promoter and the 3' UTR was introduced to separate the target gene and downstream Sh ble gene. Three different chlorella transformation vectors containing human erythropoietin (EPO) gene were constructed. The mp335EPO vector consists of a promoter from the 33kDa peptide gene, whereas the mp3353EPO vector contains the same promoter from the 33kDa peptide gene and its 3' UTR. The mp35S33pEPO vector contains the 35S promoter and the 3' UTR from the 33 kDa peptide gene. There was no significant difference in the expression levels of EPO protein in chlorella cells transformed with either of three of the transformation vectors. These data indicate that the promoters from the chlorella virus are comparable to the most common CaMV 35S promoter. Furthermore, these data suggest that other promoters from this virus can be used in future construction of chlorella transformation system for higher expression of target proteins.
Lee Chang Hyun;Han Woong;Kim Young Soo;Lee Kwang Gyu
Journal of Physiology & Pathology in Korean Medicine
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v.18
no.1
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pp.187-193
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2004
Constipation is a common clinical problem that comprises of symptoms that include excessive straining, hard feces, feeling of incomplete evacuation and infrequent defecation. Although many conditions, such as metabolic problems, fiber deficiency, anorectal problem, an drug, can cause constipation. This study was examined the effects of Chilsun-Whan on intestinal mucosa and gastrointestinal transit time and plasma lipids in rats. Adult male rats were fed for weeks on diets containing no addition(basal diet group), 5% cellulose(cellulose group) and 2.5% . Chilsun-Whan group(Chilsun-Whan group). The results were as follows; 1. The fecal weght was significantly increased 2 times in Chilsun-Whan administrated group compare to basal diet group. 2. The gastrointestinal transit times was significantly decreased in Chilsun-Whan administrated group compare to basal diet. 3. Carmine red mixed with Chilsun-Whan, as a marker, was administered through a gastric tube for stomach or intracecally by a chronically implanted catheter for colon transit. Small intestinal transit and large intestinal transit time were significantly decreased in Chilsun-Whan administrated group compare to basal diet. 4. The height of jejunal villi was developed in Chilsun-Whan administrated group compare to basal diet The thickness of mucosa and muscle layer of colonic mucosa were significantly developed in Chilsun-Whan administrated group compare to basal diet group. 5. The change of goblet cell in colonic mucosa was increased acid mucin stained alcian blue in Chilsun-Whan administrated group compare to basal diet and cellulose group. 6. HDL-cholesterol of plasma lipid was increased in Chilsun-Whan administrated group compare to basal diet and cellulose groups. Theses results suggests that Chilsun-Whan may be used in prevention and treatment of constipation resulting in increase of fecal weight, decrease of gastrointestinal transit time. development of intestinal villi, intensify of stainability of acid mucin in colon.
Adverse changes in individual's biochemistry under heavy metal stress are directly linked with its metabolic activity and health status. The present investigation highlights the differences in protecting role of Panax ginseng extract against mercuric chloride induced alterations in serum proteins. The assessment was based on dividing fifty albino rats into two sets, one for acute and the other for sub-acute study. All the sets had five groups with five albino rats in each i.e. control group, mercuric chloride treated group, Panax ginseng extract treated group, mercuric chloride followed by Panax ginseng extract treated group and Panax ginseng extract followed by mercuric chloride treated group. Mercuric chloride was given orally 0.926 mg/kg body weight for acute set and 0.044 mg/kg body weight for sub-acute set after LD50 (9.26 mg/kg body weight) determination by probitt analysis. 10 mg/kg body weight Panax ginseng extract was given in both acute and sub-acute sets after incorporating safety trials. The control group received tween-20 and distilled water only. The result exhibited significantly reduction (P<0.01) in serum protein, albumin and globulin following mercuric chloride intoxication whereas significant (P<0.01) enhancement in other groups with Panax ginseng extract as an ingredient confirming its protective role. All serum samples were also electrophoresed in 10% SDS with standard marker using discontinuous buffering system. Gradual disappearance of alpha-2 and beta-1 globulin bands from electrophoretic pattern was observed, while a single sharp band was observed between beta-2 and gamma globulin in serum protein pattern of acutely mercuric chloride treated rats. However, this band could not be visualized in sub-acute studies. Panax ginseng extract exhibits a better protection after acute intoxication.
Objectives : The purpose of this study was to evaluate effects of taping therapy on recovery of behavioral symptoms and neural excitability of the lumbar spinal cord in rat model for ankle sprain. Methods : Adult Sprague-Dawley rats was used and divided into 3 experimental groups: normal group(n=6), ankle sprain(n=6), and ankle sprain with taping treatment(n=6). In order to induce ankle sprain the right ankle joint was injured with 4~5 repetitive over-flexions and over-extensions manually. The severity of joint pain was evaluated by measuring foot weight bearing force ratio(FWBRF) of the hind limb and the injury-induced edema formation by diameter of the joint following ankle sprain. The changes of neural excitability in the lumbar spinal cord was tested by observation of cFos protein expression, a metabolic marker for neural excitation. Results : Severity of ankle injury induced in this experiment coincided with Grade 1 ankle sprain. Compared with ankle sprain group, ankle sprain+taping showed a significant reductions of joint pain as well as of edema formation at the ankle joint following ankle sprain. There was significant upregulation of cFos-immunoreactive neurons in the lumbar spinal cord 24 hours after ankle sprain. In contrast, taping therapy resulted in significant inhibition of cFos-immunoreactive neurons in the lumbar spinal cord. Conclusions : Collectively, these results suggest that taping therapy may be an alternative therapeutic intervention for symptom recovery of the mild ankle sprain.
Ji, Hye-Young;Lee, Seung-Seok;Yoo, Sung-Eun;Kim, Hosoon;Lee, Dong-Ha;Lim, Hong;Lee, Hye-Suk
Archives of Pharmacal Research
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v.27
no.2
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pp.239-245
/
2004
KR-31543, (2S,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(2 -methyl-2H-tetrazol-5-ylmethyl) amino]-3,4-dihydro-2-dimethoxymethyl-3-hydroxy-2-methyl-2H-1-benzopyran, is a new neuroprotective agent for preventing ischemia-reperfusion damage. This study was performed to identify the metabolic pathway of KR-31543 in human liver microsomes and to characterize cytochrome P450 (CYP) enzymes that are involved in the metabolism of KR-31543. Human liver microsomal incubation of KR-31543 in the presence of NADPH resulted in the formation of two metabolites, M1 and M2. M1 was identified as N-(4-chlorophenyl)-N-(2-methyl-2H-tetrazol-5-ylmethyl)amine on the basis of LC/MS/MS analysis with a synthesized authentic standard, and M2 was suggested to be hydroxy-KR-31543. Correlation analysis between the known CYP enzyme activities and the rates of the formation of M 1 and M2 in the 12 human liver microsomes have showed significant correlations with testosterone 6$\beta$-hydroxylase activity (a marker of CYP3A4). Ketoconazole, a selective inhibitor of CYP3A4, and anti-CYP3A4 monoclonal antibodies potently inhibited both N-hydrolysis and hydroxylation of KR-31543 in human liver microsomes. These results provide evidence that CYP3A4 is the major isozyme responsible for the metabolism of KR-31543 to M1 and M2.
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