• Title/Summary/Keyword: pH stimulation

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LAMP-3 (Lysosome-Associated Membrane Protein 3) Promotes the Intracellular Proliferation of Salmonella typhimurium

  • Lee, Eun-Ju;Park, Kwan-Sik;Jeon, In-Sook;Cho, Jae-Woon;Lee, Sang-Jeon;Choy, Hyun E.;Song, Ki-Duk;Lee, Hak-Kyo;Choi, Joong-Kook
    • Molecules and Cells
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    • v.39 no.7
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    • pp.566-572
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    • 2016
  • Lysosomes are cellular organelles containing diverse classes of catabolic enzymes that are implicated in diverse cellular processes including phagocytosis, autophagy, lipid transport, and aging. Lysosome-associated membrane proteins (LAMP-1 and LAMP-2) are major glycoproteins important for maintaining lysosomal integrity, pH, and catabolism. LAMP-1 and LAMP-2 are constitutively expressed in Salmonella-infected cells and are recruited to Salmonella-containing vacuoles (SCVs) as well as Salmonella- induced filaments (Sifs) that promote the survival and proliferation of the Salmonella. LAMP-3, also known as DC-LAMP/CD208, is a member of the LAMP family of proteins, but its role during Salmonella infection remains unclear. DNA microarray analysis identified LAMP-3 as one of the genes responding to LPS stimulation in THP-1 macrophage cells. Subsequent analyses reveal that LPS and Salmonella induced the expression of LAMP-3 at both the transcriptional and translational levels. Confocal Super resolution N-SIM imaging revealed that LAMP-3, like LAMP-2, shifts its localization from the cell surface to alongside Salmonella. Knockdown of LAMP-3 by specific siRNAs decreased the number of Salmonella recovered from the infected cells. Therefore, we conclude that LAMP-3 is induced by Salmonella infection and recruited to the Salmonella pathogen for intracellular proliferation.

Effect of Millimeter Waves on Quality Characteristics of Cheonggukjang (밀리미터파 처리가 청국장의 이화학적 특성에 미치는 영향)

  • Seo, Dong-Ho;Kim, Mi-Seon;Kum, Jun-Seok
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.126-131
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    • 2017
  • Millimeter waves are electromagnetic waves with frequencies of 30-300 GHz (wavelength 1-10 mm), and millimeter wave stimulation affects microorganism growth. The present study stimulated Bacillus subtilis with 60 and 70 GHz millimeter waves during cheonggukjang fermentation and characterized the effects on cheonggukjang quality. Cheonggukjang treated with millimeter wave irradiation showed no significant difference in total bacterial count but generated only 5.52-5.92% viscous substance. Irradiation with 60 GHz millimeter waves yielded bright and intense color values relative to 70 GHz millimeter waves. Examination of the amylase activity and reducing sugar content of finished cheonggukjang revealed that irradiation at 70 GHz inhibited amylase activity in cheonggukjang. Furthermore, irradiation at 70 GHz increased protease activity, whereas irradiation at 60 GHz inhibited the activity. Moreover, the amino acid content changed with millimeter wave irradiation.

Effects of Handling and Transport Vehicle on Stress and Carcass Quality of Market Pigs (출하취급과 차량형태가 출하돈의 스트레스와 도체품질에 미치는 영향)

  • Kim, Doo-Hwan;Song, Jun-Ik;Jeon, Jung-Hwan
    • Journal of Animal Environmental Science
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    • v.17 no.2
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    • pp.101-106
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    • 2011
  • One hundred and forty-four cross-bred market pigs weighing approximately 110 kg were randomly divided into four groups in a 2 (handling stress; minimal vs stimulated) ${\times}$ 2 (transport vehicle type; enclosed box vs conventional open) factorial arrangement of treatments. The stimulated handling stress group received overally rough handling including electric prod stimulation during loading, transport and lairage at least once at each step. All the animals received 3-h lairage prior to slaughter. Blood and longissimus dorsi muscle (LM) samples were taken at slaughter and after overnight chilling of the carcass, respectively. Plasma concentrations of stress indicators glucose, cortisol, creatine kinase and lactate dehydrogenase were greater in the stimulated vs minimal handling stress group. There were no interactions between the handling stress and transport vehicle type in their effects on these blood variables. The incidence of pale, soft and exudative (PSE) carcass and drip loss of LM, 24-h postmortem LM pH and color including the lightness and redness were not affected by the handling stress and transport vehicle type. In conclusion, results suggest that rough handling inflicts a stimulated stress on the animal, which is manifested by increased blood concentrations of stress indicators, and therefore should be avoided for animal welfare.

Interaction of Imipramine and $K^+$ Channel Blockers on Detrusor Muscle Strips Isolated from Canine Urinary Bladder (개의 적출방광 평활근에서 Imipramine과 $K^+$ 통로 봉쇄제와의 상호작용)

  • Huh, Joun-Young;Choi, Eun-Mee;Choi, Hyoung-Chul;Ha, Jeoung-Hee;Lee, Kwang-Youn;Kim, Won-Joon
    • The Korean Journal of Pharmacology
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    • v.31 no.2
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    • pp.195-206
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    • 1995
  • The study was undertaken to examine the possibility of the involvement of $K^+$ channels in the mechanism of relaxant-action of imipramine on the isolated canine detrusor muscle strips. Canine urinary bladder were isolated, and smooth muscle strips of 15 mm long and 2 mm wide from the mid-portion of anterior wall were made in the Tyrode solution of $0{\sim}4^{\circ}C$. The strips were prepared for isometric myography in Biancani's isolated muscle chamber containing 1 ml of Tyrode solution, which was maintained with pH 7.4 by aeration with $95%\;O_2/5%CO_2\;at\;37^{\circ}C$. RP 52891, a non-specific $K^+$ channel opener, concentration-dependently suppressed the spontaneous phasic contractions of the detrusor strips. Imipramine, a tricyclic antidepressant, also reduced the spontaneous contractions in a concentration-dependent manner. RP 52891 was more potent than imipramine(p<0.05), and Imipramine was more efficient than RP 52891(p<0.05).Procaine, a voltage-dependent $K^+$ channel blocker, glibenclamide, an ATP-dependent $K^+$ channel blocker, and apamin, a calcium-dependent $K^+$ channel blocker antagonized the relaxant effect of RP 52891, but not of imipramine. Imipramine reduced the electric field stimulation (EFS) -induced contractions concentration-dependently. None of the $K^+$ channel blockers employed for this study, procaine, glibenclamide or apamin antagonized the inhibitory action of imipramine on the EFS-induced contraction. These results suggest that in canine detrusor, the $K^+$ channels of the characteristics of voltage-dependent, ATP-dependent and/or calcium-dependent are exist, and the inhibitory action of imipramine on the contractility of the detrusor is independent from the $K^+$ channels.

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Untersuchungen iiber Blutenknospenbildung bei Ribes nigrum L. (리베스(Ribes nigrum L. var. 'Rosenthals Langtraubige Schwarze') 식물의 화아분화에 관한 연구)

  • Yang, Deok Cho
    • Journal of Plant Biology
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    • v.27 no.3
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    • pp.179-189
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    • 1984
  • Durch vergleichende Untersuchungen von Ribes nigrum L. var. 'Rosenthals Langtraubige Schwarze' zwischen $prim\"{a}rem$ und $sekund\"{a}rem$ Jugendstadium (P.J. und S.J.) eine vertiefte Einsicht in die beiden zeitlich unterschielich langen $Entwicklungsabl\"{a}ufe$ zu bekommen und auf diese Weise bessere Ansatzpunke $f\"{u}r$ die Regulierung der $Bl\"{u}tc-nknospeninduktion$ zu gewinnen, war das Ziel vorliegenden Untersuchungen. Diese Untersuchungen $f\"{u}hrten$ zu den flogenden Ergebnissen: Die Dauer der Jugendphase von Ribes nigrum L. betrug bei den P.J.-Pflanzen ca. 7 Monate, bei den S.J.-Pflanzen ca. 5 Monate. Die $Bl\"{u}hreife$-(Ripeness to flower)-setzte bei den P.J.-Pflanzen erst beim 20. Nodium ein, bei den S.J.-Pflanzen bereits beim 5. Nodium. Das $f\"{u}hrte$ dazu, da${\beta}$ bei gleicher Nodienzahl die P.J.-Pflanzen weniger $Bl\"{u}te-nknospen$ angesetzt hatten als die S.J.-Pflanzen. Die erste $Bl\"{u}tenknospendifferenzierung$ setzte bei den P.J.-Pflanzen in der oberen Triebmitte etwa am 30. Nodium ein, beiden S.J.-Pflanzen bereits in der unteren Tricbmitte etwa am 20. Nodium. Bie den P.J.-Pflanzen waren die untersten 20. Nodien immer steril, $w\"{a}hrend$ bei den S.J.-Pflanzen $h\"{a}uflgsten$ an dem ersten Nodium $\"{u}ber$ der Basis fertil waren. In allen Versuchen hat sich gezeigt, da${\beta}$ der Wachstumsabschlu${\beta}$ -(Bildung einer Terminal Knospe)-, der bei Ribes nigrum L. durch Kurztag eingeleitet wird, bei den P.J.-Pflanzen langsammer als bei den S.J.-Pflanzen erfolgt. Dies $f\"{u}hrt$ dazu, da${\beta}$ die Pflanzen der $sekund\"{a}ren$ Jugendphase auf die photokybernetischen Stimulation empfindlicher reagieren als die der $prim\"{a}ren$ Jugendphase. Alle untersuchten Versuchsdaten $f\"{u}hren$ zu dem Schlu${\beta}$, da${\beta}$ die Wurzelgibberelline $(GA_n)$ keinen Einflu${\beta}$ auf die $Jugendsterilit\"{a}t$ ($Basissterilit\"{a}t$) haben. Aus den Untersuchungen geht hervor, da${\beta}$ die induktive Ma${\beta}$nahme zur Beschleunigung der $Bl\"{u}tenknospenbildung$ $f\"{u}r$ eine $Z\"{u}chtungsselektion$ erst dann eingesetzt werden sollen, wenn die P.J.-Pflanzen mehr als 20 Nodien ausgebildet haben.

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In Vitro Development of Bovine Nuclear Transfer Embryos Reconstructed with Fetal Fibroblasts (태아 섬유아세포로 재구성된 핵치환 소 수정란의 체외발달)

  • Koo, D.B.;Choi, Y.H.;Park, J.S.;Kim, H.N.;Kang, Y.K.;Lee, C.S.;Han, Y.M.;Park, H.D.;Lee, K.K.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.407-417
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    • 2000
  • The present study was to examine effects of various electrical stimulus treatments used for electro-fusion on the preimplantation development of bovine nuclear transfer (NT) embryos with fetal fibroblast cells. Fetal fibroblast cells were isolated from one fetus at day 45 of gestation in Holstein cow, and passaged 3 to 4 times before being transferred into enucleated oocytes. Single fibroblast cells were individually placed into the perivitelline space of enucleated oocytes by using a micromanipulator. At first, the fusion and developmental rates of reconstructed oocytes were compared between different electric stimulation conditions. When fusion of the reconstructed oocyte was induced by different electric pulse periods (15, 30 and 45 $\mu$sec) at a DC pulse of 1.8 kV/cm, 15 (45.5%, 120/264) or 30 $\mu$ sec group (43.9%, 106/241) showed a higher fusion rate than 45 $\mu$sec group (23.2%, 58/250, P<0.05). However, no difference was detected in the development rate of the fused oocytes to blastocysts between groups. Next experiment was to examine the effects of different electrical field strengths (1.5, 1.8 and 2.1 kV/cm) for 15 $\mu$sec at electrofusion on in vitro development of the NT embryos. As results, there was no difference in the fusion and developmental rates of the NT embryos between electrical strength (P>0.05). Finally, developmental competence of bovine NT embryos with somatic cells was compared with IVF-derived embryos. Of enucleated oocytes fused with fibroblast cells, 27.4% (75/274) developed to the blastocyst stage, which is similar to that (24.5%, 58/237) of IVF-derived embryos. However, mean nuclei number of NT blastocysts was smaller than that of IVF-derived blastocysts. Thus, we have established an optimal condition (1.8 kV/cm, 15 $\mu$sec) for electric fusion of bovine NT oocytes with somatic cells. The present study indicates that bovine reconstructed embryos with somatic cells normally develop to blastocyst stage in vitro, although having smaller nuclei numbers of blastocysts as compared to IVF-derived embryos.

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Longitudinal flowcytometric measurement of respiratory burst activity of neutrophils in patients with pneumonia (폐렴경과 중 순환 호중구의 Respiratory Burst 활성도 변화)

  • Lee, Jae Myung;Lee, Jong Min;Kim, Dong Gyu;Choi, Jeong Eun;Mo, Eun Kyung;Park, Myung Jae;Lee, Myung Goo;Hyun, In Gyu;Jung, Ki-Suck;Park, Chan Jeoung
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.5
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    • pp.728-735
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    • 1996
  • Background : Recognition and ingestion of opsonized microorganisms by neutrophils induces the burst of oxidative metabolic activity. Products of the respiratory burst activity provide powerful oxygen dependent killing mechanism. Measurement of respiratory burst activity has been a major indicator of the functional capacity of neutrophils. We determined the respiratory burst activity of neutrophils in patients with pneumonia and observed the changes during the clinical course of pneumonia. Methods: The EDTA blood was drawn from 24 normal controls and same numbers of pneumonia patients. The respiratory burst activity(with the production of $H_2O_2$ which changes nonfluorescent DCF-DA to green fluorescent DCF) in the non-stimulated state and the stimulated state with fMLP and PMA of neutrophils was measured by flowcytometry at day 1, 3, 5, 7 and 9 of admission. Results: The respiratory burst activity of neutrophils was mildly increased by stimulation with fMLP. But there was no statistical significance between normal control and patients with pneumonia. The respiratory burst activity of neutrophils was markedly increased by stimulation with PMA in both groups. There was a significant difference in response to PMA between normal control and patients with pneumonia. The production of hydrogen peroxide from neutrophils was decreased during early course of pneumonia and it was recuperated gradually to normal level in 9 days. Conclusion : Hydrogen peroxide production from neutrophils was suppressed during early course of pneumonia and restored after treatment. It is suggested that the production of oxygen radical in response to PMA stimulation from each neutrophils is decreased rather than increased during the early course of pneumonia.

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The Role of Adenosine Receptors on Acetylcholine Release in the Rat Striatum

  • Kim, Do-Kyung;Kim, Hyeon-A;Choi, Bong-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.1-12
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    • 1997
  • As it has been reported that the depolarization induced acetylcholine (ACh) release is modulated by activation of presynaptic $A_1$ adenosine heteroreceptor and various evidence suggest that indicate the $A_2$ adenosine receptor is present in the striatum, this study was undertaken to delineate the role of adenosine receptors on the striatal ACh release. Slices from the rat striatum were equilibrated with $[^3H]$choline and then the release amount of the labelled product, $[^3H]$ACh, which was evoked by electrical stimulation (rectangular pulses, 3 Hz, 2 ms, 24 mA, $5\;Vcm^{-1}$, 2 min), was measured, and the influence of various agents on the evoked tritium outflow was investigated. And also, quantitative receptor autoradiography and drug-receptor binding assay were performed in order to confirm the presence and characteristics of $A_1$ and $A_2$ adenosine receptors in the rat striatum. Adenosine $(10{sim}100\;{mu}M)$ and $N^6$-cyclopentyladenosine (CPA, $1{sim}100\;{mu}M)$ decreased the $[^3H]$ACh release in a dose-dependent manner without changing the basal rate of release in the rat striatum. The reducing effects of ACh release by adenosine and CPA were abolished by 8-cyclopentyl-1,3-dipropy-Ixanthine (DPCPX, 2 ${mu}M$), a selective $A_1$, adenosine receptor antagonist, treatment. The effect of adenosine was potentiated markedly by 3,7-dimethyl-1-propargylxanthine (DMPX, 10 ${mu}M$), a specific $A_2$ adenosine receptor antagonist. 2-P-(2-carboxyethyl)phenethylamimo-5'-N- ethylcarboxamidoadenosine hydrochloride (CGS-21680C), in concentrations ranging from 0.01 to 10 ${mu}M$, a recently introduced potent $A_2$ adenosine receptor agonist, increased the $[^3H]$ACh release in a dose related fashion without changing the basal rate of release. These effects were completely abolished by DMPX $(10\;{mu}M)$. In autoradiograrhy experiments, $[^3H]$2-chloro-$N^6$-cyclopentyladenosine ($[^3H]$ CCPA) bindings were highly localized in the hippocampus and the cerebral cortex. Additionally, lower levels of binding were found in the striatum. However, $[^3H]$CGS-21680C bindings were highly localized in the striatal region with the greatest density of binding found in the caudate nucleus and putamen. Lower levels of binding were also found in the nucleus accumbens and olfactory tubercle. In drug-receptor binding assay, binding of $[^3H]$ CCPA to $A_1$ adenosine receptors of rat striatal membranes was inhibited by CPA ($K_i$ = 1.6 nM) and N-ethylcarboxamidoadenosine (NECA, $K_i$ = 12.9 nM), but not by CGS-21680C ($K_i$ = 2609.2 nM) and DMPX ($K_i$ = 19,386 nM). In contrast, $[^3H]$CGS-21680C binding to $A_2$ denosine receptors was inhibited by CGS-21680C ($K_i$ = 47.6 nM) and NECA ($K_i$ = 44.9 nM), but not by CPA ($K_i$ = 2099.2 nM) and DPCPX ($K_i$ = 19,207 nM). The results presented here suggest that both types of $A_1$ and $A_2$ adenosine heteroreceptors exist and play an important role in ACh release in the rat striatal cholinergic neurons.

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Studies on the Suitability and Efficiency of Human Follicular Fluid as Protein Supplement in Assisted Reproductive Technology(ART);III. Effect of Human Follicular Fluid on Improvement of Pregnancy Rates in ART (생식보조시술시 단백질원으로서 인간난포액의 적합성 및 효율성에 관한 연구;III. 인간난포액이 생식보조시술시 임신율 향상에 미치는 효과)

  • Koo, J.J.;Chi, H.J.;Kim, D.H.;Kim, J.Y.;Chang, S.S.
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.1
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    • pp.103-108
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    • 1996
  • Through the previous studies(I,II), it was observed that human follicular fluid(HFF) was more effective than human fetal cord serum(HFCS) on promoting meiotic resumption of oocytes and improving embryonic development of mouse in vitro. On the basis of these results, we have gradually exchanged HFCS with HFF as protein supplement in human ART. This study was performed to investigate the efficiency of HFF on improving the pregnancy rate in ART. Oocytes were retrieved transvaginally from patients treated with pituitary suppression with GnRH-agonist and ovarian stimulation with human menopausal gonadotro-pin(HMG) and pure follicle stimulating hormone(FSH). Aspirated oocytes were rinsed and cultured in TCM-199 containing HFF, and the concentrations of HFF were adjusted to 10, 20, and 30% according to the use for insemination, embryo growth and embryo transfer, respectively. As possible as, we tried to do embryo transfer into fallopian tube to mimic the coincidence of the cell stage with the place of sojourn in vivo, so we performed various ART programs(IVF & ET; in vitro fertilization, ZIFT; zygote intra fallopian-tube transfer, ZIFT & ET) according to the tubal conditions of patients. On the while, intra cytoplasmic sperm injection(ICSI) was used to assist IVF of the patients who had shown poor standard IVF results by immunological or severe male factor. Of the 255 cycles of ART programs using HFF as protein supplement, 118 cycles were turn out to be succeeded in pregnancy(46.2%, per cycle, p<0.05), while 21 pregnancies were achieved in the 69 cycles using HFCS(30.4%). The 255 cycles using HFF were subdivided into cycles with the type of ART programs, and each pregnancy rate of the ART programs were 44.7% (IVF & ET, 76/170 cycles), 53.4%(ZIFT, 31/58 cycles) and 40.7% (ZIFT & ET, 11/27 cycles), respectively. In the 61 ICSI cycles using HFF, 28 cycles succeed in pregnancy(45.9%), while 7 pregnancies were obtained in the 17 ICSI cycles using HFCS. Also the ongoing pregnancy rate in the group using HFF(78.8%, 93/118 cycles) was higher than that in the group using HFCS(61.9%). Therefore, we found that the use of HFF as protein supplement was more suitable and effective than the use of HFCS to improve the pregnancy rate in ART.

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Culture of tissue-cyst forming strain of Toxoplama gondii and the effect of cyclic AMP and pyrimidine salvage inhibitors (Toxoplasma gondii 약독주의 배양과 그 성장에 미치는 cyclic AMP와 pyrimidine salvage 억제제의 영향)

  • 최원영;박성경
    • Parasites, Hosts and Diseases
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    • v.32 no.1
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    • pp.19-26
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    • 1994
  • An in uipo culturing to examine the cyst stage of ToxopLQsma gondii (ME49 strain) was Investigated using murine peritoneal macrophages, and we also examined the effect of CAMP or DHFR Inhibitors on the growth of bradyzoltes. For experiments ICR mice were Injected 1.p. with 1,500 brain cysts. At 1, 3, 5 and 7 days, peritoneal exudates were Isolated and then adherent peritoneal macrophages were cultured for 1,3,5 and 10 days. Growth pattern of bradyzoltes was measured by (3H)-uracil uptake assay and morphological pattern of pseudocysts formed in macrophages was observed Uth Glemsa stain. Mostly bradyzoites were observed In the macrophages extracted at 3 and S days post Infection. After 3 days in vitro, a number of pseudocysts were formed in the macrophages and the size of pseudocysts was increased during further 5 and 10 days in vitro culture. CAMP stimulated the growth of bradyzoltes when in uiuo 3 and 5 days and then in vitro 5 and 10 days conditions were applied. In case of.DHFR Inhibitors, pynmethamlne produced a linearly decremental effect with a cont.-dependent mode but methotrexate was not effective against Intracellular bradyzoltes or pseudocysts In this system. It was suggested that cyst-forming strain of T gondii (ME49 strain) could be maintained and cultivated in uitro by use of murine peritoneal macrophages. In uivo 3 and 5 days and then in uiko 5 and 10 days conditions appeared to be suitable for culturing of bradyzoltes. CAMP and pyrimethamine had an effect of stimulation and inhibition on the growth of bradyzolte, respectively.

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