• BMB Reports
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• v.42 no.7
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• pp.418-420
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• 2009

We describe a method for producing polyclonal antibodies against peptide antigen cytochrome P450 1A2 and 3A4 using a tandem repeat of the epitope region and incorporation of proline residue between the repeated sequences. An ELISA assay revealed more efficient generation of polyclonal antibodies to tandem repeat peptide antigens than mono-epitope peptides. The incorporation of proline residues further stimulated antibody production.

• The Journal of the Korean dental association
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• v.19 no.7 s.146
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• pp.609-614
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• 1981

Effects of steroid hormones on the collagen biosynthesis in aorta and uterus were studied with ovariectomized Sprague-Dawley rats. Effects of administration of hormones, such as estrogen, testosterone and prednisolone, to the ovariectomized animals were studied, comparing with the control. Each group was injected with ³H-proline and sacrificed, followed by removals of aorta and uterus. Separations and quantitative analyses of proline and hydroxyproline were performed by means of thin layer chromatography; and radioactivities of the separated amino acids were assayed by liquid scintillation counter. Normally the incorporation of ³H-proline into hydroxyproline was greater in uterus than in aorta, and collagen turnover rate of uterus was observed rapid as well than that of aorta. In the two tissues from ovariectomized rats, the incorporation rate of ³H-proline into hydroxypoline was markedly decreased than that of the former. Changes in the turnover rate of collagen in these tissues were not observed. Decrease in ³H-proline incorporation into collagen in ovariectomized rats was markedly antagonized by estrogen, but not influenced by prednisolone in the tissues tested.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.3
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• pp.241-246
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• 1992

To clarify the requirement of L-ascorbic acid (AsA) in collagen synthesis, the incorporation of 1-$^{14}$ C-proline into the tissues of guinea pigs and the specific radioactivity ratio (proline/hydroxyproline) in collagen were investigated. Male guinea pigs maintained on the AsA-deficient diet were divided into three groups ; group A (AsA-deficient animals) : group B (control animals) supplemented with 5mg AsA/day ; group C (high dose animals) with 300mg AsA/day, and orally supplemented with or with-out AsA for 14 days. Collagen synthesis was estimated by measuring the incorporation of labeled pro-line into collagen in lung and dorsal skin, and the hydroxyproline contents in lung and skin. The AsA contents in the tissues were determined by high-peforrnance liquid chromatography (HPLC), and serum alkaline phosphatase activity was also measured. The serum alkaline phosphatase activity of AsA deficient group was very low as compared with those of AsA supplemented group. Incorporation of labelled proline into collagen and its specific radioactivity ratio in collagen increased with increasing levels of AsA in the tissues. There was a significantly positive relationship between the levels of AsA and hydroxyproline in the tissues.

• BMB Reports
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• v.31 no.2
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• pp.201-208
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• 1998

Amino acid analogs, like other inducers of stress response, induce the synthesis of stress proteins in mammalian cells. In this study, Drosophila Kc cells, in which translation is tightly controlled during stress response, was treated with proline analogs, L-azetidine-2-carboxylic acid (AzC) and 3,4-dehydro-L-proline (dh-P). Kc cells exposed to AzC or dh-P induced the synthesis of several proteins which had the same molecular weights as known heat shock proteins. However, in Kc cells, normal protein synthesis still continued in the presence of amino acids analogs unlike in heat-shocked cells. For the induction of stress response, the incorporation of dh-P into the protein was not essential, but the incorporation of AzC was. The stress protein synthesis was regulated mainly at the transcriptional level by AzC, whereas it was regulated by dh-P at the transcription level and possibly posttranscription level. During recovery, the stress protein synthesis stopped sooner in analog-treated cells than in heat-shocked cells even though the accumulated amount of Hsp70 was much less in proline analogstreated cells. It could be concluded that the proline analogs, AzC and dh-P, induced stress response through a different mechanism from heat shock.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.17 no.1
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• pp.51-87
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• 1987

The incorporation of ³H-proline by epithelial and connective tissue elements of rat palatal mucosae was studied in order to investigate the relative levels of protein synthesis by the epithelium and underlying connective tissue cells. Following a sixty minutes incorporation of the radioactive tracer in vitro, it was found that the suprabasal cells had most grains per unit area. Furthermore, the grains were more concentrated over the cytoplasm than the nucleus. This was in contrast with the labeling of basal cells which had twice as many grains over the nucleoplasm than that over the cytoplasm. In intermediate cells; i.e., the spinous layer, the number of silver grains per unit area was decreased from that of the suprabasal cells. In areas where desmosomes were more prominent, many grains were in touch with such desmosomes. However, the labeling appeared to be reduced as soon as the cells became flattened. Moreover, the epidermal keratohyalin granules were relatively free of grains. Except for certain intercellular surfaces the keratinized cells were generally free of grains. On the connective tissue side, silver grains were primarily localized over the fibroblasts with occasional grains being found over palatal muscle cells, neural elements and so on. Most grains over collagenous fibers were found in relation to mature collagen fibrils. Thus, protein synthesis in isolated mucosae of the rat palate appeared to take place both in epithelial and connective elements. There were no apparent tissue alterations caused by the in vitro incorporation procedure utilized under conditions of this study.

• The korean journal of orthodontics
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• v.20 no.2
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• pp.227-245
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• 1990

The purpose of this study was to analyze the reorganization of periodontal ligament after collagenase treatment with autoradiography. The author compared the collagenase-treated experimental group and no-treated experimental group with control group. Fourty eight Sprague-Dawley rats were divided into nine groups, including normal control and immediate group. Closed coil springs were used between the upper incisors and the first molars with 100 grams. Collagenase and $^3H-proline$ were adminstered and the samples were sacrificed and sectioned. After being dipped into the NTB-3 emulsion the samples were analyzed with light microscope under H/E stain. Data were analyzed by t-test and ANOVA. The results were as follows: 1) Generally collagenase-treated groups got more $^3H-proline$ uptake than no-treated groups. 2) Compared with normal control group, collagenase-treated group had the same $^3H-proline$ uptake in amount at 21th day. 3) Among cemento-enamel junction, middle, apex areas, cementa-enamel junction area of collagenase-treated group arrived at normal control level earlier than no-treated group. 4) Cemento-enamel junction area had the most $^3H-proline$ incorporation amount in no-treated group, but apex area had the most in collagenase group.

• Journal of Ecology and Environment
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• v.36 no.4
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• pp.255-265
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• 2013

This study was conducted to determine the extent of drought resistance based on physiological responses of Calystegia soldanella under water deficit. In order to investigate the changes of plant growth, stomatal density, photosynthesis, chlorophyll fluorescence, the contents of chlorophyll and carotenoid, osmolality, total ion contents, the contents of carbohydrate and proline, C. soldanella was grown under well watered and drought stressed conditions for 12 days. In this study, water-deficit resulted in remarkable growth inhibition of C. soldanella. The effect of water-deficit on plant growth was associated with low osmotic potential of soil. On day 12 after drought treatment, dry weight, relative water contents, number and area of leaves and stem length were lower than those of control. The stomatal conductance and net photosynthetic rate were significantly reduced in water stressed plant to regulate inner water contents and $CO_2$ exchange through the stomatal pore. Chlorophyll fluorescence and chlorophyll contents were not different in comparison with the control, indicating that the efficiency of photosystem II was not affected by drought stress. This results could be explained that water-deficit in C. soldanella limits the photosynthetic rate and reduces the plant's ability to convert energy to biomass. A significant increase in total ion contents and osmolality was observed on day 7 and day 12. Accumulation of proline in leaves is associated with the osmotic adjustment in C. soldanella to soil water-deficit. Consequently, this increase in osmolality in water stressed plant can be a result in the increase of ion contents and proline.

• Archives of Pharmacal Research
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• v.26 no.11
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• pp.929-936
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• 2003

Methanol extract (MeOH), n-hexane (Hx), chloroform ($CHCl_3$), ethyl acetate (EA), butanol (BuOH) and aqueous ($H_2O$) fractions of Eucommiae Cortex including geniposidic acid (GA), geniposide (GP) and aucubin (AU) were tested for their therapeutic efficacy on osteoporosis. The contents of GA, GP and AU in the cortex and leaf of Eucommia ulmoides Oliver were quantified by HPLC. The effect of Eucommiae Cortex on the induction of growth hormone (GH) release was studied by using rat pituitary cells. The proliferation of osteoblast-like cells increased by herbal extracts was assayed using a tetrazolium (MTT), alkaline phosphatase (ALP) activity, and [$^3H$]-proline incorporation assays. The inhibition of osteoclast was studied by using the coculture of mouse bone marrow cells and ST-2 cells. As a result, the GA, GP and AU were present in the cortex more than in the leaf of E. ulmoides Oliver. The MeOH (1mg/mL), Hx, $CHCl_3$ and EA fractions (each 20 $\mu$ g/mL) had potent induction of GH release. The $CHCl_3$ exhibited the potent proliferation of osteoblasts. The AU, GP and GA were increased proliferation of osteoblasts. In addition, GA ($IC_{50}: 4.43{\times}10^{-7}$M), AU and GP were significantly inhibited proliferation of osteoclast. In summary, it is thought that the components in a part of the fractions of Eucommiae Cortex participate in each step of mechanism for activating osteoblast to facilitate osteogenesis, and suppress osteoclast activity to inhibit osteolysis.

• The korean journal of orthodontics
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• v.23 no.2 s.41
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• pp.229-248
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• 1993

[ $Interferon-\gamma$ ] has been suggested as a cytokine of connective tissue stabilizer. In addition, it has also been demonstrated that this cytokine inhibited bone remodeling activities of the bone derived cells. In order to illuminate the effects of this cytokine in orthodontic force induced bone remodeling, it was administered to primary cultured periodontal ligament cells which have been known to have some osteoblast like characteristics. $Interferon-\gamma$ slightly decreased $[^3H]thymidine$ incorporation rate without a significant change in the total cellular DNA content up to 1000 U/ml, which meant these doses were not cytotoxic to the cell. Total protein synthesis was not influenced by various concentration of interferon-y whether it was determined by the $[^3H]proline$ incorporation rate or by the Lowry smethod. The effect of $interferon-\gamma$ on the individual protein was, however, differential, ie, it increased $[^3H]proline$ incorporation into the noncollagenous protein marginally, while it decreased $[^3H]proline$ incorporation into the collagen, so that it caused dose-dependent suppression of the relative collagen synthesis. On the contrary, the fibronectin synthesis determined by the ELISA was increased by 1000 U/ml of $interferon-\gamma$. The differential effects of the interferon-y on the collagen and fibronectin synthesis exhibited not only their protein level but also the steady state mRNA level. $Interferon-\gamma$ decreased steady state level of ${\alpha}1(I)$ procollagen mRNA significantly, while showing no significant changes in the fibronectin mRNA level. In addition to this, it was also found that indomethacin did not affect on the $interferon-\gamma$ induced collagen decrease in this cell, which meant prostaglandins were not involed in the process of $interferon-\gamma$ induced collagen decrease. So it can be concluded that the incubation of periodontal ligament cells with 1000 U/ml of $interferon-\gamma$ for 24 hr showed differential effects on the type I collagen and fibronectin gene expression. The decrease in relative collagen synthesis in the protein level was related with decrease in the steady state level of mRNA, while the increase in the fibronectin synthesis in the protein level was not correlated with the mRNA level.

• Proceedings of the SCSK Conference
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• 1996.07a
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• pp.52-67
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• 1996

새로운 노화방지 물질로 개발한 3-APPA가 노화에 의해 야기되는 여러 변화들, 특히 세포 증식, 유전자 수준 및 단백질 수준에서의 collagen의 생합성 변화, 면역조직화학염색을 이용한 collagen 생합성의 변화등을 세포배양 및 동물실험을 통하여 측정하였다. MTT assay를 이용한 인체 피부 섬유아세포의 증식 실험에서 3-APPA는 무처치군에 비교해서 최고 2배의 섬유아세포 증식 효능을 나타내었으며, $^3$[H]-proline incorporation 방법을 이용한 단층세포 배양 및 3차원 dermal equivalent 섬유아세포 배양에서 무처치군 및 vitamin C 처리군에 비해 최고 1.5배의 collagen 생합성 증가를 나타내었다. 그러나 type I alpha-procollagen mRNA expression에는 영향을 미치지 않는 것으로 나타났다. H&E 염색을 이용한 hairless mice의 피부에 대한 형태학적 변화 및 type I pM procollagen antibody를 이용한 면역조직화학염색에서, 3-APPA는 collagen 생합성을 증가시키는 것으로 나타났다. 이상의 결과에서 3-APPA는 섬유아세포 배양 및 hairless mouse를 이용한 실험에서 피부 섬유아세포 증식을 촉진시키며 collagen 생합성을 증가시켜 피부노화를 억제 할 수 있는 물질임을 밝혔다.