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Identification of Viral RNA by Light Microscopic in situ Hybridization (광학현미경 In Situ Hybridization에 의한 Viral RNA 증명)

  • 최원기;주경웅;김석홍
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.249-255
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    • 1996
  • In this paper, a in situ hybridization(ISH) has been used to investigate the yield of viral RNA expression from each organ tissues. It is studied to establish a rapidly, specific diagnostic method detecting rabbit haemorrhagic disease virus(RHDV) RNA in 10% formalin-fixed, paraffin-em-bedded tissues of naturally RHDV-infected rabbits using oligonucleotide probe to be made by RHDV total sequences. Biotin was used as the oligonucleotide probe marker. in situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method. All ISH procedure of RHDV were completed to Mi-croProbe$^{TM}$ capillary action system within 1-2 hours. In this report, RHDV was distributed widely in the cytoplasm of liver cell and the cortex of kidney but lung tissue and medulla of kidney were showed to positive reaction at locally. Although not entirely free of technical limitations, nucleic acid identification holds advantages over other diagnostic tests, including exquisite sensitivity, specificity, interchangeability and speed. It is expected that, in the immediate future viral nucleic acid detection will be a prominent part of the methods used in histopathology.

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Identification of Novel Alternatively Spliced Transcripts of RBMS3 in Skeletal Muscle with Correlations to Insulin Action in vivo

  • Lee, Yong-Ho;Tokraks, Stephen;Nair, Saraswathy;Bogardus, Clifton;Permana, Paska A.
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.301-307
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    • 2009
  • Whole-body insulin resistance results largely from impaired insulin-stimulated glucose disposal in skeletal muscle. Our previous studies using differential display and quantitative real-time RT-PCR have shown that a novel cDNA band (DD23) had a higher level of expression in insulin resistant skeletal muscle and it was correlated with whole-body insulin action, independent of age, sex, and percent body fat. In this study, we cloned and characterized DD23. The DD23 sequence is part of the 3'UTR region of the RNA binding motif, single stranded interacting protein (RBMS3). We have cloned the full length cDNA for RBMS3 and identified two splice variants. These variants named DD23-L and DD23-S have 15 and 14 exons respectively and differ from RBMS3 in the 3'UTR significantly. Northern blot analyses showed that an ~8.8 kb mRNA transcript of DD23 was predominantly expressed in skeletal muscle and to a lesser extent in placenta, but not in heart, brain, lung, liver, or kidney, unlike RBMS3. Elevated expression levels of these novel alternatively spliced variants of RBMS3 in skeletal muscle may play a role in whole body insulin resistance.

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Mechanism of Enzymatic Hydrolysis of Raw Corn Starch by Purified Glucoamylase of $\alpha$-Amylase in an Agitated Bead Reaction System (Glucoamylase 및$\alpha$-Amylase의 분쇄마찰매체 효소반응계에서의 생전분 효소분해 Mechanism)

  • 박동찬;이용현
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.260-267
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    • 1990
  • The mechanism of enzymatic hydrolysis of raw corn starch by the purified glucoamylase and a - amylase in an agitated bead reaction system was studied by investigating the changes of sugar profiles produced by each enzyme, the granular structure of raw corn starch, the amount of enzyme adsorption on residual starch, and the amylose content in residual raw starch. The sugar profiles produced by the action of exo-type glucoamylase or endo-type $\alpha$ -amylase in an agitated bead system were not recognizably differed with those produced in reaction system without bead. Without enzyme the intergenic microcrystalline structure of starch granule was not changed by the simple mechanical impact of solid media, but it was cleaved. However, starch granule was fragment into large number of small particles by the synergistic action of enzyme and attrition-milling media, identified to be the major saccharification enhancing mechanism along with the increased amount of enzyme adsorption. The amylose content decreased more readily in an agitated bead reaction system, especially by $\alpha$ -amylase.

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Involvement of D2 Receptor on Dopamine-induced Action in Interstitial Cells of Cajal from Mouse Colonic Intestine

  • Zuoa, Dong Chuan;Shahia, Pawan Kumar;Choia, Seok;Jun, Jae-Yeoul;Park, Jong-Seong
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.218-226
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    • 2012
  • Dopamine is an enteric neurotransmitter that regulates gastrointestinal motility. This study was done to investigate whether dopamine modulates spontaneous pacemaker activity in cultured interstitial cells of Cajal (ICCs) from mouse using whole cell patch clamp technique, RT-PCR and live $Ca^{2+}$ imaging analysis. ICCs generate pacemaker inward currents at a holding potential of -70 mV and generate pacemaker potentials in current-clamp mode. Dopamine did not change the frequency and amplitude of pacemaker activity in small intestinal ICCs. On the contrary dopamine reduced the frequency and amplitude of pacemaker activity in large intestinal ICCs. RT-PCR analysis revealed that Dopamine2 and 4-receptors are expressed in c-Kit positive ICCs. Dopamine2 and 4 receptor agonists inhibited pacemaker activity in large intestinal ICCs mimicked those of dopamine. Domperidone, dopamine2 receptor antagonist, increased the frequency of pacemaker activity of large intestinal ICCs. In $Ca^{2+}$-imaging, dopamine inhibited spontaneous intracellular $Ca^{2+}$ oscillations of ICCs. These results suggest that dopamine can regulate gastrointestinal motility through modulating pacemaker activity of large intestinal ICCs and dopamine effects on ICCs are mediated by dopamine2 receptor and intracellular $Ca^{2+}$ modulation.

생물방제균 Bfacillus subtilis YB-70의 외부 Urease 유전자 도입과 길항력 증강

  • Choi, Jong-Kyu;Kim, Yong-Su;Lee, Eun-Tag;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.25 no.1
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    • pp.30-36
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    • 1997
  • To genetically breed powerful multifunctional antagonistic bacteria, the urease gene of alkalophilic Bacillus pasteurii was transferred into Bacillus subtilis YB-70 which had been selected as a powerful biocontrol agent against root-rotting fungus Fusarium solani. Urease gene was inserted into the HindIII site of pGB215-110 and designated pGU266. The plasmid pGU266 containing urease gene was introduced into the B. subtilis YB-70 by alkali cation transformation system and the urease gene was very stably expressed in the transformant of B. subtilis YB-70(pGU266). The optimal conditions for the transfomation were also evaluated. From the in vitro antibiosis tests against F. solani, the antifungal activity of B. subtilis YB-70 containing urease gene was much efficient than that of the non-transformed strain. Genetic improvement of B. subtilis YB-70 by transfer of urease gene for the efficient control seemed to be responsible for enhanced plant growth and biocontrol efficacy by combining its astibiotic action and ammonia producing ability.

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Partial Purification and Properties of Non-specific $\beta$ -fructofuranosidase Produced by Bacillus subtilis (Bacillus subtilis가 생산하는 비특이적 $\beta$-fructofuransoidase의 부분정제 및 특성)

  • 송근섭;엄태붕
    • Microbiology and Biotechnology Letters
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    • v.18 no.5
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    • pp.484-489
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    • 1990
  • An intracellular inulase ( fJ-fructofuranoside fructohydrolase, EC 3.2.1.26) from Bacillus subtilis has been partially purified and its mode of action and general properties were studied. The enzyme had an apparent molecular weight of 49,000 as estimated by gel filtration and its pI point was 5.2. Substrate concentration studies showed an apparent Km of 10 mM for sucrose and of 18 mM for raffinose. The enzyme was an acid-labile protein with a pH optimum of 6.6. The optimum temperature was 50$^{\circ}$C. The enzyme acts on straight chain oligo- and poly-fructosides of the inulin series via a exo-wise cleavage mechanism, as well as on sucrose.

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Adiponectin Level in Non-Pregnant Women, Pregnant Women without Diabetes and Pregnant Women with Diabetes

  • Jo, Yoon-Kyung;Im, Jee-Aee
    • Biomedical Science Letters
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    • v.14 no.4
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    • pp.233-238
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    • 2008
  • Gestational diabetes mellitus (GDM) is a common complication during pregnancy and one of the main causes of adverse fetal-maternal outcomes. However, the pathogenesis of GDM has not been clearly stated. Adiponectin, an adipose tissue-derived plasma protein, is involved in regulation of insulin resistance and glucose hemostasis, and thus is a key modulator of insulin action and glucose metabolism. In this study, we investigated to compare serum adiponectin levels in pregnant women with diabetes, pregnant women who are without diabetes, and non-pregnant women, and to evaluate relationship between serum adiponectin. levels and metabolic parameters. Forty-one pregnant women with diabetes, fifty-nine pregnant women without diabetes and forty non-pregnancy women were recruited. Adiponectin levels were significantly lower in pregnant women with diabetes when compared to non-pregnant women and pregnant women without diabetes. Pregnant women without diabetes at second trimester had lower adiponectin levels compared to non-pregnant women. Adiponectin was negatively correlated with BMI, fasting insulin, HOMA-IR, total cholesterol, and triglyceride. In conclusion, this study confirmed that the decreased level of adiponectin precedes the onset of abnormal glucose level during pregnancy and also normal pregnant women had lower adiponectin levels compared to non-pregnant women. This knowledge may help to identify strategies for lowering the occurrence of GDM in women who are at high risk of developing the disorder.

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Effect of Baicalein on t-Butylhydroperoxide-Induced Cell Injury in Renal Tubular Epithelial Cells

  • Soon-Bee Jung
    • Biomedical Science Letters
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    • v.9 no.4
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    • pp.189-193
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    • 2003
  • This study was undertaken to investigate the effect of baicalein, a major flavone component of Scutellaria balicalensis Georgi, on oxidant-induced cell injury in renal epithelial cells. Opossum kidney cells, an established proximal tubular epithelial cells, were used as a cell model of renal epithelial cells and t-butylhydroperoxide (tBHP) as an oxidant drug model. Cell viability was measured by MTT assay and lipid peroxidation was estimated by measuring the content of malondialdehyde, a product of lipid peroxidation. Exposure of cells to tBHP caused cell death and its effect was dose-dependent over concentration range of 0.1~1.0 mM. When cells were exposed to tBHP in the presence of various concentrations (0.1~10 $\mu$M) of baicalein, tBHP-induced cell death was prevented with a manner dependent of baicalein concentration. tBHP induced A TP depletion, which was significantly prevented by baicalein. Similarly, tBHP-induced DNA damage was prevented by baicalein. tBHP produced a marked increase in lipid peroxidation and its effect was completely inhibited by baicalein. These results indue ate that tBHP induces cell injury through a lipid peroxidation-dependent mechanism in renal epithelial cells, and baicalein prevented oxidant-induced cell injury via antioxidant action inhibiting lipid peroxidation. In addition, these results suggest that baicalein may be a candidate for development of drugs which are effective in preventing and treating renal diseases.

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The Effect of Astradix-P on the Growth of Yeast for Putrefing Soy Sauce (간장 부패균의 생육에 미치는 Astradix-P의 영향)

  • 이귀주;김상달;서정훈
    • Microbiology and Biotechnology Letters
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    • v.2 no.1
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    • pp.9-11
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    • 1974
  • This experiment was carried out to conform the antseptic effect of Astradix-P, as a yeaststatic substance on the growth of film forming yeasts, which were isolated from denatured home prepared Korean soy sauce. It resulted that Astradix-P did not give any antiseptic effect, if these yeasts were inoculated into soy sauce medium, but in the ordinary medium the yeast growth were strongly inhibited. Consquently the possiblities of the practical application of the Astradix-P into the home-preserving soy sauce was primarily remained in doubt at this moment. This result might be caused from the reason that the basic amino acids, originally existed in soy sauce, eg, arginine, histidine and lysine etc., had anticipated with the antagonistic action and thereby they made the Astradix-P inactive to these yeast, which already they have been recognized in the previous works.

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Effect of mPER1 on the Expression of HSP105 Gene in the Mouse SCN

  • Kim Han-Gyu;Bae Ki-Ho
    • Biomedical Science Letters
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    • v.12 no.1
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    • pp.53-56
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    • 2006
  • The suprachiasmatic nucleus (SCN) of the anterior hypothalamus is the circadian pacemaker entrained to the 24-hr day by environmental time cues. Major circadian genes such as mPeriod ($mPer1{\sim}3$) and mCryptochrome ($mCry1{\sim}2$) are actively transcribed by the action of CLOCK/BMAL heterodimers, and in turn, these are being suppressed by the mPER/mCRY complex. In the study, the locomotor activity rhythms of mPer1 Knockout (KO) mice are measured, and the expression profiles of Heat Shock Protein 105kDa (HSP 105) genes in the SCN were measured by in situ hybridization. In agreement with previous reports, the locomotor activity rhythm of mPer1 KO mice was much shorter than that of wildtype. In addition, the total bout of activity of mPer1 KO was less in comparison to control mice. The expression of HSP 105 in the SCN of mPer1 KO mice was ranged from CT6 to CT22, with a peak level at CT14, implying that the gene are under the control of circadian clock. However, the expression of HSP 105 in the SCN of wildtype could not be detected in our study. Further analysis will reveal the direct or indirect regulation by mPer1 on the expression in the SCN and the role of the gene in the circadian clock.

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