Park, Seung-Joon;Park, Hee-Soon;Lee, Mi-Na;Sohn, Sook-Jin;Kim, Eun-Hee;Jung, Jee-Chang;Frohman, Lawrence A.;Kineman, Rhonda D.
The Korean Journal of Physiology and Pharmacology
/
v.7
no.2
/
pp.79-84
/
2003
We have previously reported that expression of the somatostatin receptor subtypes, sst1-5, is differentially regulated by growth hormone (GH)-releasing hormone (GHRH) and forskolin (FSK), in vitro. GHRH binds to membrane receptors selectively located on pituitary somatotropes, activates adenylyl cyclase (AC) and increases sst1 and sst2 and decreases sst5 mRNA levels, without significantly altering the expression of sst3 and sst4. In contrast FSK directly activates AC in all pituitary cell types and increases sst1 and sst2 mRNA levels and decreases sst3, sst4 and sst5 expression. Two explanations could account for these differential effects: 1) GHRH inhibits sst3 and sst4 expression in somatotropes, but this inhibitory effect is masked by expression of these receptors in unresponsive pituitary cell types, and 2) FSK inhibits sst3 and sst4 expression levels in pituitary cell types other than somatotropes. To differentiate between these two possibilities, somatotropes were sequentially labeled with monkey anti-rat GH antiserum, biotinylated goat anti-human IgG, and streptavidin-PE and subsequently purified by fluorescent-activated cell sorting (FACS). The resultant cell population consisted of 95% somatotropes, as determined by GH immunohistochemistry using a primary GH antiserum different from that used for FACS sorting. Purified somatotropes were cultured for 3 days and treated for 4 h with vehicle, GHRH (10 nM) or FSK ($10{\mu}M$). Total RNA was isolated by column extraction and specific receptor mRNA levels were determined by semi-quantitative multiplex RT-PCR. Under basal conditions, the relative expression levels of the various somatostatin receptor subtypes were sst2>sst5>sst3=sst1> sst4. GHRH treatment increased sst1 and sst2 mRNA levels and decreased sst3, sst4 and sst5 mRNA levels in purified somatotropes, comparable to the effects of FSK on purified somatotropes and mixed pituitary cell cultures. Taken together, these results demonstrate that GHRH acutely modulates the expression of all somatostatin receptor subtypes within GH-producing cells and its actions are likely mediated by activation of AC.
KIM Se-Kwon;LEE Hyun-Chel;BYUN He-Guk;JEON Yon-Jin
Korean Journal of Fisheries and Aquatic Sciences
/
v.29
no.2
/
pp.246-255
/
1996
To develop a natural antioxidative peptide, the gelatin was extracted from fish (Yellowfin sole) skin by hot $water(50^{\circ}C)$ extraction method and hydrolyzed with Alcalase, pronase and collagenase through a continuous 3-step membrane reactor. Each step enzymatic hydrolysates were determined the antioxidative activity and their synergistic effects, compared with $\alpha-tocopherol$ and butylated hydroxytoluene (BHT). Also, we tried to investigate the antioxidative disposition of peptide which was successfully separated by gel filtration, ion-exchange chromatography, and HPIC in cultured rat hepatocytes intoxicated with tert-butyl hydroperoxide (TBHP). Second step enzymatic hydrolysate (SSEH) among all hydrolysates and $\alpha-tocoperol$ was showed the strongest antioxidative activity. The optimum concentration of antioxidative activity for SSEH was $1\%(w/w)$ in linoleic acid. The synergistic effects were increased in using the hydrolysate with tocopherol and BHT. In the presence of the peptide isolated from SSEH, supplemented hepatocytes exposed to TBHP showed that delayed cell killing and decreased significantly the lipid peroxidation, compared with hepatocytes not cultured with isolated peptide.
Purpose: The objective of this clinical presentation was to present a clinical case series report of socket preservation, sinus augmentation, and bone grafting using a horse-derived biomaterial. Methods: A horse-derived biomaterial was used in 8 patients for different indications including socket preservation following tooth extraction, osseous bone grafting, and sinus augementation procedures. Surgeries were performed by a well trained specialist and clinical radiographs were obtained at designated intervals. Biopsy cores of 2 ${\times}$ 8 mm prior to implant placement was obtained following a healing interval of 4 - 6 months. A clinical and histologic evaluation was performed to evaluate the clinical effectiveness and biocompatibility of the biomaterial. Results: All surgeries in 8 patients were successful with uneventful healing except for one case with membrane exposure that eventually resulted with a positive outcome. Radiographic display of the healing phase during different intervals showed increased radiopacity of granular nature as the healing time increased. No signs of adverse effect or infection was observed clinically and the tissues surrounding the biomaterial seemed well-tolerated with good intentional healing. The augmented sinuses healed uneventfully suggesting in part, good biocompatibility of the biomaterial. Dental implants placed following socket preservation were inserted with high initial torque suggesting good initial stability and bone quality. Conclusions: Our results show that at least on a tentative level, a horse-derived biomaterial may be used clinically in socket preservation, sinus augmentation, bone grafting techniques with good intentional healing and positive results.
Journal of Korean Society of Environmental Engineers
/
v.37
no.12
/
pp.657-667
/
2015
Cyanobacteria frequently dominate the freshwater phytoplankton community in eutrophic waters. Cyanotoxins can be classified according to toxicity as neurotoxin (Anatoxin-a, Anatoxin-a(s), Saxitoxins) or hepatotoxin (microcystins, nodularin, cylindrospermopsin). Microcystins are present within cyanobacterial cells generally, and they are extracted by the damage of cell membrane. It has been reported that cyanotoxins caused adverse effects and they are acculmulated in aquatic oganisms of lake, river and ocean. In natural, microcystins are removed by biodegradation of microorganisms and/or feeding of predators. However, in process of water treatment, the use of copper sulfate to remove algal cells caused extraction of a mess of microcystins. Microcysitns are removed by physical, chemical and biological methods according to reports. The reduction of nutrients (N and P) inflow is basic method of prevention of cyanobacteria bloom formation. However, it is less effective than investigation because nutrients already present in the eutrophic lake. In natural lake, cyanobacteria bloom are not formed because macrophytes invade from coastal lake by eutrophication. Therefore, a coastal lake has to recover to prevent of cyanobacteria bloom formation.
The contents of bioactive and antioxidative activities (DPPH (${\alpha},{\alpha}'$-diphenyl-${\beta}$-picrylhydrazyl), free radical scavenging activity, peroxidation of linoleic acid and rat hepatocyte microsome, and Fe/Cu reducing power, tyrosinase inhibition activity) were tested by in vitro experimental models using water, hot water, ethanol and methanol extracts of leaves (ASL), roots (ASR), stems (ASS) and fruits (ASF) from Acanthopanax senticosus. Hot water extract from ASL showed the highest extraction yield (16.04%) as well as highest contents of phenolic compounds (2.67%) and flavonoids (1.43%). Major minerals were K, Ca and Mg. In oxidation in vitro models using DPPH free radical scavenging activity, Fe/Cu reducing power, $Fe^{2+}$/ascorbate-induced linoleic acid peroxidation by ferric thiocyanate and thiobarbituric acid (TBA) methods, tyrosinase inhibition activity and autooxidation of rat hepatic microsomes membrane, and antioxidative activities were strong in Acanthopanax senticosus. From these results, ASL extracts were shown to have the most potent antioxidative properties and contain the highest amounts of antioxidative compounds such as phenolic compounds and flavonoids. These results may provide the basic data to understand the biological activities of bio-active materials derived from leaves of Acanthopanax senticosus.
Rice bran protein hydrolysates were prepared and some of their physicochemical properties were investigated to utilize rice bran as starting material for functional food ingredient. Rice bran proteins (RBP) were prepared from defatted rice bran by alkaline extraction and isoelectric precipitation. The enzyme for hydrolysis of RBP was selected through measuring relative activity by pH-drop method and comparing the degree of hydrolysis (DH) of hydrolysates. The enzymatic hydrolysates prepared by $Esperase^{\circledR}$ treatment were partitioned into two fractions by ultrafiltration(UF) with a 10 kDa molecular weight cut-off membrane. Each fraction was applied to a cholic acid-conjugated ${\omega}-aminohexyl$ Sepharose 4B column and the bile acid-binding components were obtained by eluting with deoxycholate. Gel permeation chromatography on a Sephadex G-50 column revealed that molecular weight of the bile acid-binding fraction of UF permeate was distributed in ranges of $2\;kDa{\sim}10\;kDa$ and $0.2\;kDa{\sim}0.6\;kDa$. Three peaks (R-1, R-2 and R-3) were obtained by prep-HPLC of bile acid-binding fraction of UF retentate and analyzed for total and free amino acid composition. The results showed that proline content of the bile-acid binding polypeptides and peptides was four times as much as that of rice bran protein and that the peak corresponding to higher average hydrophobicity had a higher free amino acid content. Average hydrophobicity slightly increased with enzymatic hydrolysis.
Bovine brucellosis, an important zoonosis, is diagnosed with serological tests such as the RBT, TAT using inactivated whole bacterial cells or bacterial lipopolysaccharide (LPS) antigen in Korea. However, a strong cross-reaction between Brucella spp. and Yersinia enterocolitica O9 in these tests has seriously complicated the diagnosis of animal brucellosis because Brucella spp. shares common antigenic determinants with Y. enterocolitica O9 in the smooth LPS region. In this study, Brucella-field strains were isolated from Brucellapositive Hanwoo in Kimhae, Korea and outer membrane protein (omp) which has low cross-reaction with Y. enterocolitica O9 and high immunogenicity was extracted from the field strains Then we compared ELISA using the extract with RBT-TAT. Fifteen field strains were isolated from 47 supra-mammary-lymph nodes, which were collected from 18 farms. Isolation rate was 32%. Brucella-specific antigen was identified by performing SDS-PAGE or Western blotting on extracted omp with at 0.5% n-lauroylsarcosine One hundred and ninety-two serum-samples were used in the experiment: 142 negative and 50 positive samples verified by RBT-TAT. According to ELISA results, 127 samples were negative and 15 appeared positive among 142 negatives by RBT-TAT, while 42 samples were positive and 8 were negative among 50 positives by RBT-TAT. Therefore, it showed 89.4% of specificity and 84% of sensi-tivity. Through the current experiments, we could set up an ELISA based on the omp which has low cross-reaction and high immunogenicity and concluded that the omp could be a good material for accurate diagnosis of bovine brucellosis.
Bee pollen is produced by honeybees and is considered one of the most balanced and nourishing nutritional supplements available. Historically, bee pollen has been prescribed for its healing properties and consumed for its high-energy supply. Recent research has provided evidence that bee pollen has diverse biological activities, such as anti-oxidant, anti-inflammatory, anti-bacterial, and even anti-cancer effects. However, the outer membrane of the pollen grain, exine, is highly resistant to most acidic solutions, high pressure, and even digestive enzymes, and the resulting low bioavailability limits its nutritional and clinical applications. This study applied a wet-grinding method to destroy the exine effectively, and it then examined the pollen's enhanced biological activity. First, microscopic observations provided strong evidence that wet grinding destroyed the exine time-dependently. In addition, the content of polyphenols, well-known ingredients of bee pollen and used as internal standards for the quality control of commercial pollen preparations, increased up to 11-fold with wet grinding. Further, the anti-oxidant activity demonstrated on the ABTS anti-oxidant assay, as well as the DPPH radical scavenging assay, was also dramatically increased. Together, the results presented here support a new technology by which bee pollen can be used as a resource for medical, nutritional, and cosmetic applications.
Kim, Soo-Byeong;Chung, Kyung-Yul;Jeon, Mi-Seon;Shin, Tae-Min;Lee, Yong-Heum
Korean Journal of Acupuncture
/
v.31
no.2
/
pp.66-78
/
2014
Objectives : The specificity of acupuncture point has been a highly controversial subject. Existing researches said that ion-distribution differences are observed on the acupuncture point. This study was conducted under the assumption that multiple ionic changes induced by muscle fatigue would be different between the acupuncture point with non-acupuncture point. Methods : To induce the identical fatigue, twenty subjects performed the knee extension/flexion exercise using the Biodex System 3. ST32 and ST33 as well as adjacent non-acupuncture points were selected. We measured blood lactate and analyzed the median frequency(MF) and peak torque. To obtain the information on the extracellular fluid(ECW), intracellular fluid(ICW) and cell membrane indirectly, we used the multi-frequency bioelectrical impedance analysis(MF-BIA) method. Results : MF, peak torque and blood lactate level of all measurement sites were gradually returned to normal. Re resistance of ST32 had a stronger response, but a non-acupuncture point adjacent to ST33 had a larger response up to 20 minutes post exercise. Ri resistances were similar for both acupoints and non-acupoints. The $C_m$ capacitance of ST32 had a stronger response after inducing fatigue, but ST33 had a smaller response than a non-acupuncture point adjacent to it. Conclusions : In comparison with before and after inducing fatigue, the specificity of acupuncture points was not clearly observed. Hence, we concluded that the body composition factors extraction method had the limitation as a method of finding the specificity of acupuncture points by inducing fatigue.
Kim Sang Hui;Hong Young-Sook;Lee Jinseon;Son Dae-Soon;Lim Yu-Sung;Song In-Seung;Lee Hye-Sook;Kim Do Hun;Kim Jingook;Choi Yong Soo
Journal of Chest Surgery
/
v.38
no.1
s.246
/
pp.38-43
/
2005
Matrix metalloproteinase-2 (MMP-2) is a class of proteolytic enzymes that digest collagen type IV and other components of the basement membrane. It plays a key role in the local invasion and the formation of distant metastases by various malignant tumors. The aim of this study was to evaluate the activity of MMP-2 and its significance as a prognostic marker in resected stage I non-small cell lung cancer (NSCLC). Material and Method: In this study we obtained fresh-frozen samples of tumor and non-tumor tissues from 34 patients with stage I NSCLC who underwent resection without preoperative radiotherapy or chemotherapy. After the extraction of total protein from tissue samples, MMP-2 activities were assessed by gelatin-substrate-zymography. The activities were divided into the higher or lower groups. Result: The MMP-2 activities were higher in tumor tissues than in non-tumor tissues. The MMP-2 activity of non-tumor tissues in recurrent group was higher than in non-recurrent group (p<0.01). Also the patients with higher MMP-2 activity of non-tumor tissues showed poor 5 year survival (p<0.01). Conclusion: This result indicates that the higher level of MMP-2 activity in the non-tumor tissue is associated with the recurrence and survival after the resection of stage I NSCLC. Therefore, MMP-2 activity in the non-tumor tissue could be used as a potential prognostic marker for the resected stage I-NSCLC.
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