• Parasites, Hosts and Diseases
• /
• v.27 no.2
• /
• pp.87-100
• /
• 1989

Eimeria tenella, an intracellular protozoan parasite infecting the epithelial cells of the ceca of chickens, causes severe diarrhea and bleeding that can lead its host to death. It is of interest that 2. tenezla first penetrate into the mucosal intraepithelial Iymphocytes (IEL) before they parasitize crypt or villous epithelial cells. This in vitro study was undertaken to know whether the penetration of E. tenella into such a lymphoid cell is a beneficial step for the parasite survival and development. Three sequential experiments were performed. First, the in vitro established bovine kidney cell line, MDBK cells, were evaluated for use as host cells for E. tenella, through morphological observation. Second, the degree of parasite development and multiplication in MDBK cells was quantitatively assayed using radioisotope labelled uracil ($^3H-uracil$) . Third, the E. tenella sporozoites viability was assayed after preincubation of them with thicken spleen cells. E. tenella oocysts obtained from the ceca of the infected chickens were used for the source of the sporozoites. Spleen cells (I) obtained from normal chickens (FP strain) were preincubated with the sporozoites (T) at the E:T ratio of 100:1, 50:1 or 25:1 for 4 or 12 hours, and then the mixture was inoculated into the MDBK cell monolayer. Morphologically the infected MDBK cells revealed active schisogonic cycle of E. tenella in 3~4 days, which was characterized by the appearance of trophozoites, and immature and mature schizonts containing merogoites. The 3H-uracil uptake by E. tenella increased gradually in the MDBK cells, which made a plateau after 48~60 hours, and decreased thereafter. The uptake amount of $^3H-uracil$ depended not only upon the inoculum sixte of the sporozoites but also on the degree of time delay (preincubation; sporozoites only) from excystation to inoculation into MDBK cells. The 3H-uracil uptake became lower as the preincubation time was prolonged. In comparison, after preincubation of sporozoites with spleen cells for 4 or 12 hours, the 3H-uracil uptake was significantly increased compared with that of control group. From the results, it was inferred that, although the penetration of E. tenella sporozoites into the lymphoid cells such as IEL is not an essential step, it should be at least a beneficial one for the survival and development of sporozoites in the chicken intestine.

• The Journal of the Acoustical Society of Korea
• /
• v.39 no.1
• /
• pp.16-23
• /
• 2020

In High-Intensity Focused Ultrasound (HIFU) treatment, effective localization of HIFU focus is important for developing a safe treatment plan. While Magnetic Resonance Imaging guided HIFU (MRIgHIFU) can visualize the ultrasound path during the treatment for localizing HIFU focus, it is challenging in ultrasound imaging guided HIFU (USIgHIFU). In the present study, a real-time ultrasound beam visualization technique capable of localizing HIFU focus is presented for USIgHIFU. In the proposed method, a short pulse, with the same center frequency of an imaging ultrasound transducer below the regulated acoustic intensity (i.e., Ispta < 720 mW/㎠), was transmitted through a HIFU transducer whereupon backscattered signals were received by the imaging transducer. To visualize the HIFU beam path, the backscattered signals underwent dynamic receive focusing and subsequent echo processing. From in vitro experiments with bovine serum albumin gel phantoms, the HIFU beam path was clearly depicted with low acoustic intensity (i.e., Ispta of 94.8 mW/㎠) and the HIFU focus was successfully localized before any damages were produced. This result indicates that the proposed ultrasound beam path visualization method can be used for localizing the HIFU focus in real time while minimizing unwanted tissue damage in USIgHIFU treatment.

• The Korean Journal of Pharmacology
• /
• v.9 no.1
• /
• pp.59-63
• /
• 1973

Although numerous drugs are available for the treatment of superficial fungi infections of skin, and yet the clinical effects of most of such drugs are not satisfactory. In the hope of searching for effective drugs for superficial fungi infections, the authors observed fungistatic effects of Pyrethri Flos, a common herb in Korea, with water extract (PFWE), ethanol extract (PFEE), and methanol extract (PFME) from Pyrethrum cinerariaefolium V. In in vitro studies, the spores of fungi were inoculated on Sabouraud's glucose agar media which contained Pyrethri Flos extracts in each concentrations of $500\;{\mu}g/ml$, $1,000\;{\mu}g/ml$ and $5,000\;{\mu}g/ml$ respectively, and the growth of the fungi was observed for 3 weeks. The species of the fungi used in these experiments were Epidermophyton floccosum, Microsporum canis, Microsporum nanum, Microsporum gypseum, Microsporum cookei, Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans and Trichophyton verrucosum. The results of these studies were as follows; 1. The growth of M. nanum & T. rubrum was slightly inhibited by PFWE $500\;{\mu}g/ml$, and the growth of M. nanum, M. cookei & T. rubrum was slightly inhibited by PFWE $1,000\;{\mu}g/ml$. The growth of E. floccosum, M. gypseum & M. cookei was slightly inhibited, however the growth of M. canis, M. nanum, T. mentagrophytes, T. rubrum & T. tonsurans was significantly inhibited by PFWE $5,000\;{\mu}g/ml$. With $500\;{\mu}g/ml$ of PFEE, the growth of M. canis, M. nanum, T. mentagrophytes, T. rubrum & T. tonsurans was significantly inhibited, and moderate inhibition of M. cookei growth and slight inhibition E. floccosum & M. gypseum were observed. The growth of M. canis, M. nanum, T. mentagrophytes, T. rubrum & T. tonsurans was significantly inhibited, and the growth of E. floccosum, M. gypseum & M. cookei was moderately inhibited by PFEE $1,000\;{\mu}g/ml$. Significant inhibitions of the growth of E. floccosum, M. canis, M. nanum, M. gypseum, M. cookei, T. mentagrophytes, T. rubrum & T. tonsurans were observed by PFEE $5,000\;{\mu}g/ml$. 3. The growth of E. floccosum & M. cookei was moderately inhibited, and the growth of M. canis, M. nanum, M. gypseum, M. mentagrophytes, T. rubrum & T. tonsurans was significantly inhibited by PFME $500\;{\mu}g/ml$. But the growth of E. floccosum, M. canis, M. nanum, M. gypseum, M. cookei, T. mentagrophytes, T, rubrum & T. tonsurans was significantly inhibited, and the growth of T. verrucosum was slightly inhibited in both PFME $1,000\;{\mu}g/ml$ and $5,000\;{\mu}g/ml$.

• The Korean Journal of Zoology
• /
• v.30 no.1
• /
• pp.1-9
• /
• 1987

Cyclic AMP (cAMP) was known to play a key role in the regulation of cumulus expansion and oocyte maturation of mammalian cumulus-oocyte complexes (COC's) in vivo and in vitro. The present experiments were conducted to know how intracellular level of cAMP in these cells is controlled. Intracellular cAMP level was modulated by culturing mouse CGC's with an adenylate cyclase stimulator, forskolin, a phosphodiesterase inhibitor, 3-isobutyl-1-methyixanthine (IBMX), human chorionic gonadotrophin (HCG), or follicle stimulating hormone (FSH). The rate of cumulus expansion and germinal vesicle break-down (GVBD) was checked after culture and used as a biological end point. Forskolin in the medium began to stimulate the expansion of the complexes at 1 nM and induced maximum expansion (80~90%) at 0 1~10 $\mu$M. The expansion rate was reduced to 60% when forskolin concentration was increased to 100 $\mu$M. Oocyte GVBD occurred normally (75~82%) in the presence of 10 $\mu$M of forskolin, but partial suppression was appeared at 100 pM of the drug (40%). IBMX also stimulated the expansion from the concentration of 0.01 pM and induced full expansion (81~89%) between the concentration of 1-1000 $\mu$M. Meiotic resumption was occurred normally under 10 $\mu$M of IBMX, but suppressed drastically from the concentration of 100 $\mu$M. The minimum exposing time to hormone or drugs required to trigger cumulus expansion was two minutes with HCG, 15~30 minutes with FSH and fors kolin, and two hours with IBMX. The data presented here seemed to imply that intracellular cAMP level in cumulus cells is regulated by both adenylate cyclase and phosphodiesterase and cumulus expansion is induced by a peak of cAMP while meiotic arrest is maintained by continuous presence of cAMP.

• Journal of Korean Ophthalmic Optics Society
• /
• v.12 no.2
• /
• pp.79-86
• /
• 2007

The aim of the present study was to determine mechanisms of corneal epithelial cell apoptosis in vitro following exposure to anti-FAS and anti-FAS ligand antibody and during infection with mycoplasma sp.. A cultured human corneal epithelial(HCE) cell line was treated with anti-FAS antibody or anti-FAS ligand antibody for 2 and 4 days. The original cell line was found to be contaminated by mycoplasma removal agent(MRA) was used to eliminate the bacterium from the cell line. MRA($0.5{\mu}{\ell}$ tissue culture medium) was added to the cell line and incubated for 1 week. The cell line underwent multiple passages in media not contaminating MRA and cells were grown to 50-80% confluency on coverslips and stained using the Hoechst stain provided in the kit to ensure mycoplasma removal. Apoptosis experiments were performed before and after mycoplasma removal. The apoptotic index of anti-FAS and anti-FAS ligand antibody on mycoplasma contaminated cell line was studied using Hoechst 33342 staining and Annexin V-FITC and Propidium Iodide Staining. In conclusion, anti-FAS antibody induces apoptosis in HCE cells in a time and concentration-dependent mechanism. Cell lines contaminated with mycoplasma have an incresed susceptibility to FAS induced apoptosis.

• The Journal of Korean Obstetrics and Gynecology
• /
• v.27 no.2
• /
• pp.46-58
• /
• 2014

Objectives: This study was designed to investigate intestinal immune system activation and anti-metastatic effect on cancer cells by orally administered extracts of Boyanghwano-tang. Methods: To observe immunomodulating effects of Boyanghwano-tang on Peyer's patch cells, we measured cytokines GM-CSF, IL-4. In addition to observing effects of Boyanghwano-tang on hematopoiesis, we measured proliferation of bone marrow cells mediated by Peyer's patch cells in vitro. IgA induction activated in intestinal content and serum was measured to observe the effect of orally administered Boyanghwano-tang on mucosal immune system. After administering ovalbumin (OVA) with Boyanghwano-tang, Proliferation of Peyer's patch cell was measured to investigate gut immunostimulatory effect. Anti-metastatic experiments were conducted in vivo mouse model by using colon 26-M3.1 carcinoma cell. Results: The amounts of GM-CSF and IL-4 in the culture supernatant of Peyer's patch cells were significantly increased compared to the control group. The proliferation of bone marrow cell was significantly up-regualted with Boyanghwano-tang. These results indicate that oral administration of Boyanghwano-tang enhances the secretion of hematopoietic growth factors such as GM-CSF and IL-4 from Peyer's patch cells, and these cytokines also act on modulator of bone marrow cell proliferation. After orally administering OVA with Boyanghwano-tang, IgA induction and Proliferation of peyer's patch cell was up-regulated with Boyanghwano-tang. These results means orally administered Boyanghwano-tang activates intestinal immune system and has an inhibitory effect on tumor metastasis. In addition, We found that orally administered Boyanghwano-tang significantly inhibited tumor metastasis in vivo. Conclusions: Orally administered Boyanghwano-tang appears to have considerable activity on the anti-metastasis by activation of immune system.

• Asian-Australasian Journal of Animal Sciences
• /
• v.22 no.7
• /
• pp.993-1004
• /
• 2009

Three experiments were conducted to determine the effects of non-ionic surfactant (NIS) supplementation on ruminal fermentation, nutrient digestibility and performance of beef steers fed high-roughage diets. The objective of experiment 1 was to investigate the effects of NIS supplementation on in vitro ruminal fermentation of cultures administered with corn and barley as grain substrate and rice straw and timothy hay as roughage substrate. The in vivo ruminal fermentation, nitrogen balance and digestibility of nutrients were also examined with steers fed a high-roughage diet in experiment 2. The aim of experiment 3 was to determine the responses to NIS of growing steers fed a high-roughage diet. In experiment 1, ammonia nitrogen concentration for NIS supplementation was higher (p<0.05) than for the control with all substrates. However, concentrations of total volatile fatty acid (VFA), acetate, butyrate and valerate of the incubated roughage substrates, rice straw and timothy hay, were higher (p<0.05) for NIS supplementation than for the control whereas VFA concentrations in the cultures of corn and barley were unaffected. These results indicated that effects of NIS on ruminal fermentation are diet dependent, specifically on roughage sources. In experiment 2, ruminal pH of steers supplemented with NIS was lower (p<0.05) than the control. Ruminal concentrations of ammonia nitrogen, acetate, total VFA and urinary concentrations of purine derivatives were increased (p<0.05) by NIS supplementation. In experiment 3, supplementation of NIS increased (p<0.05) intakes of total feed and corn silage, average daily gain, and feed efficiency of growing steers although they varied depending on supplementation level. Due to the roughage-specific feature of NIS effects, NIS appears to enhance ruminal fermentation of fibrous parts of feeds and, consequently, performance of steers fed a high-roughage diet.

• International Journal of Oral Biology
• /
• v.33 no.4
• /
• pp.179-186
• /
• 2008

Several lines of evidence suggest that osteocytes play a critical role in bone remodeling. Both healthy and apoptotic osteocytes can send signals to other bone surface cells such as osteoblasts, osteoclasts, osteoclast precursors, and bone lining cells through canalicular networks. Osteocytes responding to mechanical strain may also send signals to other cells. To determine the role for osteocytes an mechanical strain in bone remodeling, we examined the effects of fluid flow shear stress on osteoclast precursor cell and osteoblast proliferation and recruitment induced by osteocytes. In addition, the effects of fluid flow shear stress on osteocyte M-CSF, RANKL, and OPG mRNA expression were also examined. MLO-Y4 cells were used as an in vitro model for osteocytes, RAW 264.7 cells and MOCP-5 cells as osteoclast precursors, and 2T3 cells as osteoblasts. MLO-Y4 cells conditioned medium (Y4-CM) was collected after 24h culture. For fluid flow experiments, MLO-Y4 cells were exposed to 2h of pulsatile fluid flow (PFF) at 2, 4, 8, $16{\pm}0.6\;dynes/cm^2$ using the Flexcell $Streamer^{TM}$ system. For proliferation assays, MOCP-5, RAW 264.7, and 2T3 cells were cultured with control media or 10-100% Y4 CM. Cells were cultured for 3d, and then cells were counted. RAW 264.7 and 2T3 cell migration was assayed using transwells with control media or 10-100% Y4-CM. M-CSF, RANKL and OPG in MLO-Y4 mRNA expression was determined by semiquantitative RT-PCR. Y4-CM increased osteoclast precursor proliferation and migration, but decreased 2T3 cell proliferation and migration. CM from MLO-Y4 cells exposed to PFF caused decreased RAW 267.4 cell proliferation and migration and 2T3 migration compared to control Y4-CM. However, Y4-CM from cells exposed to PFF had no effect on 2T3 osteoblastic cell proliferation. PFF decreased RNAKL mRNA and increased OPG mRNA in MLO-Y4 cells compared to control(without PFF). PFF had no effect on M-CSF mRNA expression in MLO-Y4 cells. These results suggest that osteocytes can regulate bone remodeling by communication with osteoclast precursors and osteoblasts and that osteocytes can communicate mechanical signals to other cells.

• Archives of Pharmacal Research
• /
• v.29 no.10
• /
• pp.859-865
• /
• 2006

With an approach to study the anti-tumor effects and mechanism of selenium compound, we investigated the anti-tumor activity and mechanism of $Na_5SeV_5O_{18}{\cdot}3H_2O$ (NaSeVO) in K562 cells. The results showed that $0.625{\sim}20\;mg/L$ NaSeVO could significantly inhibit the proliferation of K562 cells in vitro in a time- and concentration-dependent manner as determined by microculture tetrazolium (MTT) assay, the IC50 values were 14.41 (4.45-46.60) and 3.45 (2.29-5.22) mg/L after 48 hand 72 h treatment with NaSeVO respectively. In vivo experiments demonstrated that i.p. administration of 5, 10 mg/kg NaSeVO exhibited an significant inhibitory effect on the growth of transplantation tumor sarcoma 180 (S180) and hepatoma 22 (H22) in mice, with inhibition rate 26.8% and 58.4% on S180 and 31.3% and 47.4% on H22, respectively. Cell cycle studies indicated that the proportion of G0/G1 phase was increased at 2.5 mg/L while decreased at 10 mg/L after treatment for 24, 48 h. Whereas S phase was decreased at 2.5-5 mg/L and markedly increased at 10 mg/L after treatment for 48 h. After treatment for 24 h, 10 mg/L NaSeVO also markedly increased S and G2/M phases. Take together, the result clearly showed that NaSeVO markedly increased S and G2/M phases at 10 mg/L. The study of immunocytochemistry showed that the expression bcl-2 is significantly inhibited by 10 mg/L NaSeVO, and bax increased. Morphology observation also revealed typical apoptotic features. NaSeVO also significantly caused the accumulation of $Ca^{2+}$ and $Mg^{2+}$, reactive oxygen species (ROS) and the reduction of pH value and mitochondrial membrane potential in K562 cells as compared with control by confocal laser scanning microscope. These results suggest that NaSeVO has anti-tumor effects and its mechanism is attributed partially to apoptosis induced by the elevation of intracellular $Ca^{2+}$, $Mg^{2+}$ and ROS concentration, and a reduction of pH value and mitochondria membrane potential (MMP).

• Journal of Biomedical Engineering Research
• /
• v.19 no.4
• /
• pp.385-391
• /
• 1998

A new balancing method of atrial pressures balancing for the moving actuator total artificial heart(TAH) without an extra compliance chamber was developed. The asymmetric operation of the pendulous moving actuator have made it possible to compensate the left and right pump output difference by utilizing the interventricular air space as an internal compliance chamber in a pump housing. Furthermore, the balancing performance between left and right pump outputs is increased through the improvement of the flexibility of part of the polyurethane housing. However, the increase of the flexibility of the pump housing causes a little loss of the cardiac output due to the reduction of active filling property. In this paper., a good condition between the balance and pump output performance is evaluated by adjusting the air volume in the interventricular space through a series of in vitro experiments. This new pump was implanted in a sheep weighting 63kg, and it survived for 3 days and the average cardiac output during postoperative days was about 4.2 L/mim with the atrial pressures under 15 mmHg.