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Identification of Bovine Pregnancy-Specific Whey Proteins using Two-Dimensional Gel Electrophoresis

  • Han, Rong-Xun;Choi, Su-Min;Kim, Myung-Youn;Quan, Yan Shi;Kim, Baek-Chul;Diao, Yun Fei;Koqani, Reza;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.32 no.4
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    • pp.255-261
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    • 2008
  • The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield-the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy-specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non-pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non-pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH $3.0{\sim}10.0$ strip, by loading a 2-mg milk protein sample. After the second-dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non-pregnant and pregnant cattle milk protein spots, using ImageMaster, this was followed by analysis with MALDI TOF-MS. Analysis of the 2-DE gel image resulted in a total of approximately $500{\sim}600$ protein spots, of which 12 spots were differentially expressed, six spots were up-regulated, and four spots were down-regulated; two spots were identified as pregnancy-specific proteins. These proteins were identified as lactoferrin, NA-DH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2-D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

Survey of Multi Residual Pesticides in Materials of Korean Traditional Herbal Tea (한방차 원료의 잔류농약 조사연구)

  • Kim, Nan-Young;Kim, Young-Sug;Kim, Myung-Gil;Jung, Hong-Rae;Kim, Yun-Sung;Kim, Han-Taek;Lee, Sun-Woo;Chae, Kyeng-Suk;Yoon, Mi-Hye
    • The Korean Journal of Pesticide Science
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    • v.16 no.1
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    • pp.28-34
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    • 2012
  • We analysed 149 samples of Korean traditional herbal tea materials. The 156 pesticides were analyzed by GC/ECD and NPD, detected pesticides were confirmed by GC-TOF/MS. Sample preparation was performed bv multi-residue analysis method of multiclass pesticides of the Korea Food Code. The residual pesticides were detected in 22 samples(14.8%), the highest detection frequency samples are lycium and jujube. Detected pesticides in Korean traditional herbal tea materials were chlorpyrifos (5 samples), chlorothalonil (3 samples), cypermethrin (3 samples), hexaconazol (3 samples) and cyhalothrin(3 samples). The pesticide types detected in Korean traditional herbal tea materials were organophosphorus(29.2%), pyrethroids(16.7%), organochlorines (12.5%) and triazoles(12.5%). The 5 samples(lycium, jujube, chrysanthemum, balloon-flower, milk vetch root) were detected pesticides below MRLs, 2 samples(cornus fruit, cnidium) were detected pesticides unnotificated MRLs.

Effect of $K^+-channel$ Blockers on the Muscarinic- and $A_1-adenosine-Receptor$ Coupled Regulation of Electrically Evoked Acetylcholine Release in the Rat Hippocampus

  • Yu, Byung-Sik;Kim, Do-Kyung;Choi, Bong-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.2
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    • pp.147-154
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    • 1998
  • It was attempted to clarify the participation of $K^+-channels$ in the post-receptor mechanisms of the muscarinic and $A_1-adenosine$ receptor- mediated control of acetylcholine (ACh) release in the present study. Slices from the rat hippocampus were equilibrated with $[^3H]$choline and the release of the labelled products was evoked by electrical stimulation (3 Hz, 5 V/cm, 2 ms, rectangular pulses), and the influence of various agents on the evoked tritium-outflow was investigated. Oxotremorine (Oxo, $0.1{\sim}10\;{\mu}M$), a muscarinic agonist, and $N^6-cyclopentyladenosine$ (CPA, $1{\sim}30\;{\mu}M$), a specific $A_1-adenosine$ agonist, decreased the ACh release in a dose-dependent manner, without affecting the basal rate of release. 4-aminopyridine (4AP), a specific A-type $K^+-channel$ blocker ($1{\sim}100\;{\mu}M$), increased the evoked ACh release in a dose-related fashion, and the basal rate of release is increased by 3 and $100\;{\mu}M$. Tetraethylammonium (TEA), a non-specific $K^+-channel$ blocker ($0.1{\sim}10\;{\mu}M$), increased the evoked ACh release in a dose-dependent manner without affecting the basal release. The effects of Oxo and CPA were not affected by $3\;{\mu}M$ 4AP co-treatment, but 10 mM TEA significantly inhibited the effects of Oxo and CPA. 4AP ($10\;{\mu}M$)- and TEA (10 mM)-induced increments of evoked ACh release were completely abolished in Ca^{2+}-free$ medium, but these were recoverd in low Ca^{2+}$ medium. And the effects of $K^+-channel$ blockers in low Ca^{2+}$ medium were inhibited by $Mg^{2+}$ (4 mM) and abolished by $0.3\;{\mu}M$ tetrodotoxin (TTX). These results suggest that the changes in TEA-sensitive potassium channel permeability and the consequent limitation of Ca^{2+}$ influx are partly involved in the presynaptic modulation of the evoked ACh-release by muscarinic and $A_1-adenosine$ receptors of the rat hippocampus.

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Effects of $CO_2$ Enrichment on the Differentiation and Growth in tissue culture of Panax ginseng C. A. Meyer (人參(인삼) 조직배양(組織培養)에 있어 $CO_2$ 처리(處理)가 식물체(植物體) 분화(分化) 및 생장(生長)에 미치는 영향(影響))

  • Chung, Chan-Moon;Bae, Kil-Kwan;Aoki, Masatoshi
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.1
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    • pp.14-20
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    • 2000
  • This experiment was conducted to investigate the effects of length of storage period under low temperature, $CO_2$ enrichment and addition of plant growth regulators in Murashige and Skoog medium on the plant regeneration of Korean ginseng (Panax ginseng C. A. Meyer). Seeds were treated for 60 and 80 days respectively under $5^{\circ}C$ environment. 2500ppm of $CO_2$ was enriched by ventilation. Three plant growth regulators added to the medium were Indolbutyric acid, Benzyladenin and Gibberellic acid (GA3). The result indicated that : The capacity of differentiation was higher in the aged cotyledons from the seeds treated for 80 days under low temperature condition than in those treated for 60 days. $CO_2$ enrichment had stimulating effects on the growth and development of shoot primordium significantly but less effects on the formation of adventitious buds. From one zygotic embryo hundreds of plantlets were differentiated. $CO_2$ enrichment had effects on the formation of both indirect somatic embryo and direct somatic embryo. Indirect somatic embryo showed little growth and differentiation, being undifferentiated vascular stele and epicotyl. Direct somatic embryos were formed on the epidermis of backside basal part of cotyledon. Those embryos developed to whole plant having latent bud.

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The Study on the Analgesic Effect and its Cholinergic Mechanism of Electroacupuncture in the Rat Model of Collagen-induced Arthritis (Collagen 유발(誘發) 관절염(關節炎) 동물모델에 대(對)한 전침자극(電鍼刺戟)의 진통효과(鎭痛效果) 및 그 기전(機轉)에 관(關)한 연구(硏究))

  • Baek, Yong-hyeon;Hong, Seong-hun;Yang, Hyung-in;Park, Dong-suk;Choi, Do-young
    • Journal of Acupuncture Research
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    • v.21 no.2
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    • pp.115-129
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    • 2004
  • Objectives : To investigate the analgesic effect and its cholinergic mechanism of electroacupuncture(EA) in the rat model of collagen-induced arthritis(CIA). Methods : Immunization of male Sprague-Dawley rats with bovine typeII (CII) collagen emulsified in Freund's incomplete adjuvant, followed by a booster injection 14 days later, leads to development of arthritis in more than 70% of rats by 21 days postinjection. After three weeks of first immunization, EA stimulation(2 Hz, 0.07 mA, 0.3 ms) was delivered into Jogsamni($ST_{36}$) for 30 minutes. Analgesic effect was evaluated by tail flick latency(TFL). We compared the analgesic effect of EA with TFLs between pretreatment of normal saline and pretreatment of Atropine (1 mg/kg, intraperitoneal) and Neostigmine ($100{\mu}g/kg$, intraperitoneal) in CIA. Results : 1. TFLs were gradually decreased in CIA as increasing severity of arthritis. 2. Jogsamni($ST_{36}$) EA stimulation in CIA increased TFLs and the effect lasted for 60 minutes. 3. Increased TFLs with Jogsamni($ST_{36}$) EA stimulation were inhibited with pretreatment of atropine in CIA 4. Increased TFLs with Jogsamni($ST_{36}$) EA stimulation did not show an obvious synergistic effect with pretreatment of neostigmine in CIA. Conclusions: Jogsamni($ST_{36}$) EA showed analgesic effects in CIA. The analgesic effects of Jogsamni($ST_{36}$) EA were inhibited by atropine pretreatment and combined application of Jogsamni(ST36) EA and neostigmine did not show an synergistic effect. These observations suggest that intrinsic muscarinic cholinergic pathways represent an important modulating system in pain perception of inflammatory pain in CIA It is suggested that, the active mechanism of analgesic effect in EA may involve the release of acetylcholine in the spinal cord.

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Ameliorating Effects of the Cognitive-Enhancing Korean Herbs on Neurotoxic-Induced Amnesia in Rats (새로운 제형의 치매치료제제의 효능연구)

  • Kim Ji Hyun;Jung Jin Yong;Chae Yoon Byung;Hahm Dae Hyun;Park Yang Jin;Lee Hye Jung;Shim Insop
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.2
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    • pp.303-310
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    • 2002
  • Ancient Korean physicians have used several oriental herbs to cure dementia and these effects were described in the Korean herbal books. Some cognitive-enhancing oriental herbs have been widely used as a herbal medicine against dementia. However, few of studies have proved their efficacy in treatment for dementia. In the present study, we investigated the effects of herbal compounds, which are mainly consisted of Uncaria sinensis, Corydalis yanhusuo and Acorus gramineus on learning and memory in Mortis water maze task and the central cholinergic system of the rats with neurotoxic medial septum lesion. In water maze test, the animals were trained to find a platform in a fixed position during 6 days and then received a 60-s probe trial in which the platform was removed from the pool on the 7th day. Ibotanic and 192 saporin lesion of the medial septum (MS) impaired the performance of maze test and degenerated choline acetyltransferase (ChAT) in the brain, which is a marker of the central cholinergic system. Daily administrations of herbal medicine (100mg/kg, p.o.) for 21 consecutive days produced significant reversals of the neurotoxic-induced deficit in learning and memory. These treatments also reduced the loss of cholinergic immunoreactive neurons in the brain induced by neurotoxin. These results demonstrated that herbal compounds ameliorated learning and memory deficits through effects on the central nervous system, partly through effect on the acetylcholine system. Our studies suggest an evidence of these herbs as treatment of Alzheimer's disease.

The Analgesic Effect and Its Neuropathologic Changes of Pulsed Radiofrequency Lesions in the Sciatic Nerve of the Rat (백서 좌골신경에 시행한 박동성 고주파술 (Pulsed Radiofrequency)이 급성 통증과 신경조직에 미치는 영향)

  • Lee, Kee-Heon;Shin, Keun-Man;Kweon, Kyoung-Seok;Jung, Bae-Hee;Lim, So-Young;Hong, Soon-Yong;Choi, Young-Hee;Park, Young-Euy
    • The Korean Journal of Pain
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    • v.13 no.2
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    • pp.149-155
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    • 2000
  • Background: Pulsed radiofrequency (RF) lesioning is a painless procedure and causes no neurodestruction and neuritis-like reaction are common following conventional RF lesioning. There is little data about the effect of pulsed RF especially with regard to its suitability for the treatment of acute pain. The possibility of a placebo effect cannot be ruled out because a double-blind study was not performed in previous studies. There is also no neuropathologic study about pulsed RF. Methods: The rats were anesthetized with sodium pentobarbital (40 mg/kg, i.p.; supplemented as necessary). The common sciatic nerve was exposed by blunt dissection through biceps femoris. Pulsed RF was administered to the common sciatic nerve using a 30 ms/s pulse with for 120 seconds. The temperature reached was no more than $42^{\circ}C$. Analgesia was determined using hot-plate assay shortly and, 3 days and 1 week before, and 2 weeks after operation. Lesions were examined with LM (light microscope) and EM (electron microscope) 2 weeks later. Results: There were no differences in response latencies between the control and experimental group. There were many vacuoles with hyaline bodies in the Schwann cell cytoplasm rather than axon in LM and larger electron dense bodies. No changes were found in the axon or unmyelinated fibers. Only small changes were found in the sheaths of myelinated fibers and Schwann cells. Conclusions: We therefore do think that any analgesic effect of pulsed RF is not a result of block of neural conduction. But rather than it can be attributed to others factors. It was also ineffective as a treatment for acute pain such as that caused by the hot-plate test.

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The Anti-inflammatory Effect of Cinnamomi Ramulus (계지의 항염 효과에 관한 연구)

  • Park Hi-Joon;Lee Ji-Suk;Lee Jae-Dong;Kim Nam-Jae;Pyo Ji-Hee;Kang Jun-Mo;Choe Il-Hwan;Kim Su-Young;Shim Bum-Sang;Lee Je-Hyun;Lim Sabina
    • The Journal of Korean Medicine
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    • v.26 no.2 s.62
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    • pp.140-151
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    • 2005
  • Objectives: Cinnamomi Ramulus (CR), the young twig of Cinnamomum loureirri nees, has been used for treating symptoms related to pain, rheumatic arthritis and inflammation in Korean herb medicine. This study was carried out to investigate the anti-inflammatory effect of CR in vivo and in vitro. Methods: Extracts of CR were prepared and the chemical components of the extracts were examined by gas chromatography-mass spectrometry (GC-MS). The extracts were administrated to the rat paw edema model induced by carrageenan to evaluate the anti-inflammatory effect of CR. The expressions of nitric oxide (NO), prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 were also quantified in lipopolysaccharide(LPS)­induced RAW 264.7 macrophages to survey the effect of CR in vitro. The main components were cinnamaldehyde and coumarin. Results: We examined the anti-inflammatory activity of the $80\%$ ethanol extract of Cinnamomi Ramulus in vivo by using carrageenan-induced rat paw edema model. Maximum inhibition of $54.91\%$ was noted at the dose of l1000mg/kg after 2 hours of drug administration in carrageenan-induced rat paw edema and this showed a potent anti-inflammatory effect. Conclusions: The results showed that Cinnamomi Ramulus suppressed dose-dependently LPS-induced NO production in RAW 264.7 macrophages and also decreased iNOS protein expression. Cinnamomi Ramulus also showed a significant inhibitory effect in LPS-induced PGE2 production and COX-2 expression.

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Assessment of VOCs Emission Characteristics from Building Materials such as Wall Paper, Paints, and Adhesives Using Small Chamber Method (소형챔버법을 이용한 건축자재 중 벽지, 페인트 및 접착제의 VOCs 방출특성 평가)

  • Lee Suk-Jo;Jang Seong-Ki;Cho Yong-Sung;Jung Kyung- Mi;Jeong Gi-Ho
    • Journal of Korean Society for Atmospheric Environment
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    • v.21 no.2
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    • pp.191-204
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    • 2005
  • Building and furnishing materials and consumer product are important sources of volatile organic compounds(VOCs) and other aldehydes in the indoor environment. Some available evidence indicates that VOCs can cause adverse health effects to the building occupants and contribute to some of the symptoms of what we call, 'Sick House Syndrome' in Korea. The aims of this study were to evaluate the efficiency of emission system and to investigate comparison of the emission characteristics of different building materials such as wall-papers, paints, and adhesives. The emission of VOCs from building materials were determined in the small chambers defining the temperature, relative humidity, and ventilation rate in this study. VOCs were sampled for 20 minutes using Tenax-TA tubes and analysed by GC-MS with thermal desorption. The stability of conditions for temperature and relative humidity in this small chamber system showed that the fluctuation of temperature was between 25.4$\pm$0.3$^{\circ}C$ and that of relative humidity was 50.2$\pm$0.6$\%$ under the airflow rate of 167 mL/min. The emission tests from building materials resulted in TVOC emission rates of 0.011 $\~$ 3.108 mg/m$^{2}$h after 7 days. The general wall-papers emitted toluene abundantly and the natural wall-papers mainly emitted n-butanol and a minor amount of alkanes compound such as n -tetradecane. The remainder consisted of toluene, m,p -xylene, and styrene. The paints mainly emitted toluene and the adhesives mainly emitted chloroform as well as toluene. As a result, this study is expected to suggest meaningful data for future studies in exposure control through selecting healthy building materials and for the establishment of guidelines for various building materials in Korea.

Astaxanthin Biosynthesis in Transgenic Arabidopsis by Using Chyb Gene Encoding β-Carotene Hydroxylase (β-Carotene Hydroxylase 관련 Chyb 유전자를 이용한 형질전환 Arabidopsis에서 Astaxanthin의 생합성)

  • Lee, Ho-Jae;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.231-237
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    • 2004
  • Oxycarotenoids are oxygenated carotenoids that perform critical roles in plants. $\beta$-Carotene hydroxylase adds hydroxyl groups to the $\beta$-rings of carotenes and has been cloned from several bacteria and plants including Arabidopsis. This study was carried out to investigate the effect of $\beta$-carotene hydroxylase gene (Chyb) on the oxycarotenoids biosynthesis in the transgenic Arabidopsis. Construct of pGCHYB containing Chyb was established onto Gateway vector system (pENTR3C gateway vector and pH2GW7 destination vector). Arabidopsis thaliana (cv. Columbia) was transformed with Agrobacterium tumerfacience GV3101 harboring pGCHYB construct driven by 35S promoter and hygromycin resistant gene. Seven hundred bases paired PCR products, indicating the presence of Chyb gene, were found in the transformants by PCR analysis using Chyb primers. Hygromycin resistance assay showed that transgenes were stably inherited to next generation. The overexpression of the Chyb gene resulted in the decrease carotenoid content. Especially, astaxanthin unusual oxycarotenoid in wild type Arabidopsis was detected in the transgenic plants. This means that decreased carotenoids might be converted into astaxanthin metabolism with the aid of silent gene in the host.