• Clinical and Experimental Pediatrics
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• v.51 no.8
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• pp.879-885
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• 2008

Purpose : The human lung fibroblast may act as an immunomodulatory cell by providing pro-inflammatory cytokines and chemokines, which are important in airway remodeling. Vascular endothelial growth factor (VEGF) induces mucosal edema and angiogenesis. Thymus and activation regulated chemokine (TARC) induces selective migration of T helper 2 cells. We investigated whether human lung fibroblasts produced VEGF and TARC, and the effects were augmented with the co-culture of fibroblasts and human bronchial smooth muscle cells (HBSMC), and whether dexamethasone can inhibit the proliferation and the release of VEGF in lung fibroblasts. Methods : Human lung fibroblasts were cultured with and without HBSMC, growth-arrested in serum-deprived medium, and pretreated with dexamethasone for 16 hours. After 24-hour stimulation with platelet derived growth factor-BB (PDGF-BB) and/or transforming growth factor-${\beta}$ (TGF-${\beta}$), culture supernatant was harvested for assays of VEGF and TARC. Cell proliferation was assayed using BrdU cell proliferation ELISA kit. Results : 1) The release of VEGF was significantly increased after stimulation with TGF-${\beta}$, and its release was augmented when co-stimulated with PDGF and TGF-${\beta}$. 2) VEGF release induced by PDGF or TGF-${\beta}$ was inhibited by dexamethasone. 3) There was no synergistic effect on the release of VEGF when human lung fibroblasts were co-cultured with HBSMC. 4) Dexamethasone did not suppress human lung fibroblasts proliferations. 5) Neither TGF-${\beta}$ nor PDGF induced TARC release from lung fibroblasts. Conclusion : Human lung fibroblasts may modulate airway remodeling by release of VEGF, but they have no synergistic effects when co-cultured with HBSMC. Dexamethasone suppresses VEGF release, not proliferation of lung fibroblast.

• Restorative Dentistry and Endodontics
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• v.20 no.1
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• pp.328-341
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• 1995

I. Shear Bond Strength to Air-dried and Remoistened Dentin.. The effect of air-drying and remoistening of acid-conditioned dentin before priming with the primer of All-Bond 2(BISCO. INC., U. S. A.) on shear bond strength(SBS) was investigated. Ninty freshly extracted sound human molars were divided at random into 9 groups of 10 teeth each. SBSs were meaured for acid-conditioned and non-conditioned dentin to which the primer and bonding agent of All-Bond 2 and composite resin(Z-100, 3M Dental Products, U. S. A.) were applied. The following values(Mean${\pm}$ SD, MPa) were obtained for the groups conditioned with 10% phosphoric acid for 15 seconds: Group l(blot dried) $6.7{\pm}4.1$ ; Group 2(10 seconds dried) $16.1{\pm}5.3$ ; Group 3(20 seconds dried) $15.4{\pm}4.8$ ; Group 4(30 seconds dried) $15.2{\pm}6.3$ ; Group 5(10 seconds dried/remoistened) $26.4{\pm}2.6$ ; Group 6(20 seconds dired/remositened) $22.2{\pm}2.7$ ; Group 7(30 seconds dried/remoistened) $21.5{\pm}4.1$. For the non-conditioned groups the values were: Group 8 (blot dried) $13.3{\pm}2.6$ ; Group 9(10 seconds dried) $12.9{\pm}3.5$. The data were analyzed using ANOVA. In the acid-conditioned groups, mean values of SBS for the air-dried specimens(Grps. 2, 3 and 4) and the 20 and 30 seconds dried/remoistened specimens (Grps. 6 and 7) were significantly lower than that of blot dried specimens.(p<0.05) The value for 10 seconds dried/remoistened specimens (Grp. 5), however, was not statistically different compared to that of blot dried specimens.(p>0.05) In the non-conditined groups, there was no statistical difference between blot dried and 10 seconds dried specimens.(p>0.05) The results suggest that the acid-conditioned dentin surface is more vulnerable to dentin bonding when it is air-dried or even remoistened after long period of drying. II. Shear bond stengh to the moistened and primed enamel. The effect of moistening and priming of enamel compared to the air-drying of enamel on the shear bond strength of enamel bonding agent was investigated. The experiment was divided into 4 groups each containing 10 caries-free maxillary incisor teeth. Shear bond strength values were measured for the primed and non-primed enamel to which All-Bond 2 and Z-100 were applied. The following values(MPa) were obtained for the primed groups pretreated with 32 % phosphoric acid for 15 seconds. : Group 1 (10 seconds dried) $29.8{\pm}2.2$ ; Group 2(moistened) $26.8{\pm}5.4$. For the non-primed groups the values were: Group 3(10 seconds dried/primed) $27.6{\pm}5.0$ ; Group 4(mostened/primed) $28.2{\pm}3.5$. The data were subjected to statistical analysis using ANOVA. The results showed that mean shear bond strengths among the experimental groups were not statistically different. (p>0.05) Conclusively, It is suggested that the bonding ability to enamel is not decreased by the moistening and priming of the enamel.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.3
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• pp.366-372
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• 2006

This study is to investigate the physicochemical properties of differently pretreated chestnut starches during starch isolation and to examine their gelatinization properties by both heat and alkali treatments. One kind is starch A made by alkali method from peeled chestnut. The other is starch B made from chestnut with the outer layer. The results are as follows. Starch A has higher water binding capacity of 86.9% than starch B with 80.66%. Swelling powers of both starch A and B increased rapidly from $60^{\circ}C\;to\;80^{\circ}C$ in both, and since then it has changed a bit. Both began to show their solubility at $60^{\circ}C$ and increased continuously as the temperature went up. Starch A has higher swelling power and solubility than starch B. In iodine reaction, starch A has higher ${\lambda}max$ and absorbance at ${\lambda}max$ than starch B. X-ray diffraction patterns showed that starch A is type $C_b$ and that starch B is type B. Starch B has higher relative crystallinity of 37.0% than starch A with 36.2%. The results by differential scanning calorimetry revealed that starch A gelatinized from $66.95^{\circ}C$ to $77.5^{\circ}C$ and its enthalpy is 2.04 cal/g. And starch B gelatinized from $67.09^{\circ}C\;to\;77.5^{\circ}C$, and its enthalpy is 2.29 cal/g. Amylograms of chestnut starch at 6.5% concentration indicated that starch B needs higher onset temperature when beginning to gelatinize than starch A does. But starch A shows much higher peak viscosity, breakdown and setback than starch B does. Starch A shows higher viscosity, gel volume, and optical transmittance in gelatinization properties by alkali than starch B does.

• Analytical Science and Technology
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• v.25 no.1
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• pp.76-82
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• 2012

The determination and confirmation of dutasteride in human serum was performed by a liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Beclomethasone as an internal standard (I.S.) was added to the serum and the mixed sample was pretreated by liquid-liquid extraction (LLE) with methyl tert-butyl ether (MTBE). The mass transitions of dutasteride and I.S. monitored in multiple reaction monitoring (MRM) were m/z 529.6${\rightarrow}$461.5 and m/z 409.3${\rightarrow}$391.2, respectively, and the retention times were 6.45 and 5.46 min, respectively. The calibration curve was linear in the concentration range of 0.5~30.0 ng/mL ($R^2$= 0.9999) and the limit of quantitation (LOQ) was found to be 0.5 ng/mL. The recovery of dutasteride was shown to be 66~72%. The intra-day assay precision and accuracy were in the range 3.5~5.5% and 85.7~89.9%, respectively, and the interday assay precision and accuracy were in the range 4.2~5.8% and 90.8~95.8%, respectively.

• The Korean Journal of Pharmacology
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• v.32 no.1
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• pp.113-123
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• 1996

It has been well known that polyamines ensure the stability of chromatin structure and the fidelity of DNA transcription. This study was carried out to evaluate the effect of polyamines on the apoptosis of mouse thymocytes induced by dexamethasone and polyamine synthesis inhibitors. 1) In the histological death findings of thymocytes double-stained with acridine orange and ethidium bromide, the apoptotic and the necrotic fractions (AF; NF) in the control group were $9.4{\pm}4.2%$ and $4.5{\pm}5.3%$, respectively. Dexamethasone $(3\;{\times}\;10^{-8}\;M:\;DX)$ in creased AF upto $52.0{\pm}8.1%$ and did not change NF, but A23187 $(5\;{\times}\;10^{-7}\;M:\;A2)$ increased AF and NF upto $45.0{\pm}8.9%$ and $20.5{\pm}10.6%$, respectively. 2) The thymocyte viability was significantly reduced by DX, DHEA $(1\;{\times}\;10^{-4}\;M)$, A2, DFMO $(1\;{\times}\;10^{-4}\;M)$, and $MGBG\;(1\;{\times}\;10^{-4}\;M)$, respectively. It was, however, little affected by $aminoguanidine\;(1\;{\times}\;10^{-4}\;M:\;AG)$, $putrescine\;(1\;{\times}\;10^{-5}\;M:\;PT)$, $spermidine\;(1\;{\times}\;10^{-5}\;M:\;SD)$, and $spermine\;(1\;{\times}\;10^{-5}\;M:\;SM)$. 3) The genomic DNA of mouse thymocyte was markedly fragmented by DX and A2, respectively, and to a lesser extent, by DHEA, but was little affected by MGBG, DFMO, AG, and each of polyamines. 4) The DX induced reduction of thymocyte viability was moderately attenuated by DHEA, but little affected by DFMO, MGBC, and AG. However, SM significantly attenuated the viability reduction induced by A2 as well as DX. 5) The thymocyte viability reduction by MGBG and DFMO was significantly attenuated by only SM among three polyamines applied in this study. 6) The thymocyte viability redution by combined treatments of DX with DFMO and MGBG, respectively, was significantly attenuated by SM, and moderately by PT. But the viability reduction by combined treatment of DX with AG or DHEA was not affected by polyamines. These results suggest that polyamines, particularly spermine, might play the inhibitory role in thymocyte apoptosis and the inhibitory effect can be ascribed in part to the increase of polyamine uptake by thymocytes pretreated with DFMO and MGBG.

• Applied Microscopy
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• v.24 no.2
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• pp.78-92
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• 1994

Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

• Journal of Preventive Medicine and Public Health
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• v.27 no.1 s.45
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• pp.11-24
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• 1994

The studies reported here were undertaken to investigate the effects of mercury chloride on immune system of Balb/c mouse employing a flexible tier of in vitro and in vivo assays. Mercury chloride inhibited the proliferative responses of spleen cells to lipopolysaccharide, pokeweed mitogen, and phytohemagglutinin as a dose-dependent manner. This inhibitory effect was observed not only when $HgCl_2$ was added 2nd or 3rd day of 3 days culture period but also when spleen cells was pretreated with $HgCl_2$ for 2 hours. Mercury chloride, however, potentiated the production of IgM and IgG from spleen cells. During the $HgCl_2$ administration by drinking for 3 weeks, the weight gain of mice was significantly blunted than that o control group mice, while no overt signs related to mercury toxicity were noted in any mice of experimental group. There was no change in thymus and spleen weights, and in histological findings of kidney, bone marrow of femur, thymus, spleen, and popliteal lymph node after 3 weeks of mercury exposure. However, $HgCl_2$ induced a significant increase of total serum IgM, IgG including $IgG_1,\;IgG_{2a}\;and\;IgG_{2b}$, and IgE in Balb/c mice. Treatment in vivo with anti-IL-4 monoclonal antibody significantly abrogated the $HgCl_2$-induced increase in total serum IgG1 and IgE. Whereas $HgCl_2$ potentiated total serum IgM and IgG, there was no difference in total serum hemagglutinin to SRBC (Sheep Red Blood Cell) between experimental and control group mice when these mice were immunized with SRBC. All these findings observed in Balb/c mice suggest that mercury perturbates well-orchestrated regulation of immune responses before developing histopathological changes in lymphoid tissues.

• Journal of Chest Surgery
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• v.37 no.1
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• pp.11-18
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• 2004

Background: We tested the effect of indomethacine and total spinal anesthesia on the improvement of placental flow during cardiopulmonary bypass on fetal lamb. Material and Method: Twenty fetuses at 120 to 150 days of gestation were subjected to bypass via trans-sternal approach with a 12 G pulmonary arterial cannula and 14 to 18 F venous cannula for 30 minutes. All ewes received general anesthesia with ketamine. In all the fetuses, no anesthetic agents were used except muscle relaxant. Ten served as a control group in which placenta was worked as an oxygenator during bypass (Control group). The remainder worked as an experimental group in which pretreatment with indomethacine and total spinal anesthesia was performed before bypass with the same extracorporeal circulation technique as control group (Experimental group). Observations were made every 10 minutes during a 30-minute bypass and 30-minute post bypass period. Result: Weights of the fetuses ranged from 2.2 to 5.2 kg. In Control group, means of arterial pressure decreased from 44.7 to 14.4 mmHg and means of Pa$CO_2$ increased from 61.9 to 129.6 mmHg at each time points during bypass. Flow rate was suboptimal (74.3 to 97.0 $m\ell$/kg/min) during bypass. All hearts fibrillated immediately after the discontinuation of bypass. On the contrary, in Experimental group, means of arterial pressure reamined higher (45.8 to 30 mmHg) during bypass (p<0.05). Means of Pa$CO_2$ were less ranging from 59.8 to 79.4 mmHg during bypass (P<0.05). Flow rates were higher (78.8 to 120.2 $m\ell$/kg/min) during bypass (p<0.05). There were slower deterioration of cardiac function after cessation of bypass. Conclusion: In this study, we demonstrated that the placental flow was increased during fetal cardiopulmonary bypass in the group pretreated with indomethacine and total spinal anesthesia. However, further studies with modifications of the bypass including a creation of more concise bypass circuit, and a use of axial pump are mandatory for the clinical application.

• Journal of Life Science
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• v.26 no.12
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• pp.1422-1430
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• 2016

This study was performed to investigate the modulatory effects of two prototypes of Panax ginseng saponin fractions, 20(S)-protopanaxadiol saponins (PDS) and 20(S)-protopanaxatriol saponins (PTS), on the induction of inflammatory mediators in lipopolysaccharide (LPS)-treated RAW264.7 murine macrophage cells. For this purpose, RAW264.7 cells were treated with LPS ($10{\mu}g/ml$) before, after, or simultaneously with PDS or PTS ($150{\mu}g/ml$), and the released level of nitric oxide (NO) and expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were evaluated. When RAW264.7 cells were treated with LPS and ginseng saponin fractions simultaneously for 24 hr, PTS, compared to PDS, more strongly attenuated the NO production induced by LPS treatment. When the cells were pretreated with LPS for 2 hr followed by PDS or PTS treatment for 24 hr, both ginseng saponins strongly reduced NO release. The pretreatment of RAW264.7 cells with PDS or PTS for 2 hr followed by LPS treatment for 24 hr significantly attenuated the LPS-induced production of NO. PTS showed stronger inhibitory potency to NO generation than PDS. Our western blot experiment showed that both PDS and PTS ($150{\mu}g/ml$) also significantly down-regulated the expressions of iNOS and COX-2 induced by LPS treatment. Our results suggest that both PDS and PTS possess strong protective effects against LPS-stimulated inflammation and that their protective effects are mediated by the suppression of NO synthesis via down-regulation of pro-inflammatory enzymes, iNOS, and COX-2 in the RAW264.7 cells.

• Journal of Embryo Transfer
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• v.28 no.1
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• pp.19-24
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• 2013

This study assesses of efficiency of oocyte recovery and in vitro development for during the non breeding season in goat. Thirty-four matured goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injections of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotropin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes were activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at $38.5^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $O_2$, 90% $N_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I+II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytes. The clavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes. There was no significant difference in oocyte quality between seasonal treatments.