• Restorative Dentistry and Endodontics
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• v.23 no.1
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• pp.316-327
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• 1998

This study was designed to examine the tissue levels of interleukin-$1{\alpha}$(IL-$1{\alpha}$), interleukin-$1{\beta}$(IL-$1{\beta}$) and tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) in inflamed human dental pulps and periapical lesions, and to determine the relationship between each cytokine and pulpal and periapical pathosis. The pulps used in this experiment, were obtained in routine endodontic treatment and the periapical lesions in periapical surgery after clinical diagnoses were performed. These specimens were divided into four groups as normal pulp group(control group, n=9), acute pulpitis group(n=g), chronic pulpitis group(n= 10) and periapical lesion group(n= 18) and stored in liquid N2. For extract preparation, tissues were finely minced with a scalpel, and the fragments were incubated in $0.5m\ell$ homogenizing buffer (0.1 mol/L potassium chloride, 0.02 mol/L TRIS; pH=7.6) for two hours and grinded with glass homogenizer. Debris was removed by centrifugation and supernatants were immediately tested with enzymelinked immunosorbent assay (ELISA, R&D Co., Minneapolis, USA). Following results were obtained; 1. The concentrations of IL-$1{\alpha}$ in all experimental groups were significantly higher than in control group(p<0.05). And the concentrations of IL-$1{\alpha}$ in periapical lesion group were somewhat higher than in two pulpitis groups, but the differences among those groups were not stastically significant (p>0.05). 2. The concentrations of IL-$1{\beta}$ in all experimental groups were significantly higher than in control group (p<0.05), and all the experimental groups expressed similar concentrations. 3. The concentrations of TNF-${\alpha}$ in all experimental groups were higher than in control group but only the differences between chronic pulpitis group and control group were statistically significant(p<0.05). And the concentrations of TNF-${\alpha}$ in acute and chronic pulpit is groups were higher than in periapical lesion group but only the differences between chronic pulpitis group and periapical lesion group were statistically significant (p<0.05). 4. There was significant correlation only between IL-$1{\alpha}$ and IL-$1{\beta}$ in periapical lesion group (p<0.05).

• Journal of The Korean Dental Society of Anesthesiology
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• v.14 no.2
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• pp.101-106
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• 2014

Background: Remifentanil, an ultra-short-acting mu-opioid receptor agonist, is unique from other opioids because of its esterase-based metabolism, minimal accumulation, and very rapid onset and offset of clinical action. Remifentanil can prevent the inflammatory response and can suppress inducible nitric oxide synthase expression in a septic mouse model. However, the effects of remifentanil on human keratinocyte and autophagy have yet to be fully elucidated during hypoxia-reoxygenation. Here we investigated whether remifentanil confers protective effect against hypoxia-reoxygenation in human keratinocyte and, if so, whether autophagy mediates this effect. Methods: The human keratinocytes were cultured under 1% oxygen tension. The cells were gassed with 94% $N_2$, and 5% $CO_2$ and incubated for 24 h at $37^{\circ}C$. To determine whether the administration of affects human keratinocytes hypoxia-reoxygenation injury, cells were then exposed to various concentrations of remifentanil (0.01, 0.1, 0.5 and 1 ng/ml) for 2 h. After remifentanil treatment, to simulate reoxygenation and recovery, the cells were reoxygenated for 12 h at $37^{\circ}C$. Control group did not receive remifentanil treatment. Normoxia group did not receive hypoxia and remifentanil treatment for 36 h. 3-MA group was treated 3-methyladenine (3-MA) for 1h before remifentanil treatment. Cell viability was measured using a quantitative colorimetric assay with MTT, showing the mitochondrial activity of living cells. Cells were stained with fluorescence and analyzed with Western blot analysis to find out any relations with activation of autophagy. Results: Prominent accumulation of autophagic specific staining MDC was observed around the nuclei in RPT group HaCaT cells. Similarly, AO staining, red fluorescent spots appeared in RPT group HaCaT cells, while the Normoxia, control and 3-MA groups showed mainly green cytoplasmic fluorescence. We here examined activation of autophagy related protein under H/R-induced cells by Western blotting analysis. Atg5, Beclin-1, LC3-II (microtubule-associated protein 1 light chain 3 form II) and p62 was elevated in RPT group cells. But they were decreased when autophagy was suppressed by 3-MA (Fig. 5). Conclusions: Although the findings of this study are limited to an in vitro interpretation, we suggest that remifentanil may have a beneficial effect in the recovery of wound from hypoxia-reoxygenation injury.

• Journal of Dental Anesthesia and Pain Medicine
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• v.16 no.3
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• pp.175-184
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• 2016

Background: This study investigated the effect of remifentanil pretreatment on Cos-7 cells exposed to oxidative stress, and the influence of remifentanil on intracellular autophagy and apoptotic cell death. Methods: Cells were divided into 4 groups: (1) Control: non-pretreated cells were incubated in normoxia (5% $CO_2$, 21% $O_2$, and 74% $N_2$). (2) $H_2O_2$: non-pretreated cells were exposed to $H_2O_2$ for 24 h. (3) RPC+$H_2O_2$: cells pretreated with remifentanil were exposed to $H_2O_2$ for 24 h. (4) 3-MA+RPC+$H_2O_2$: cells pretreated with 3-Methyladenine (3-MA) and remifentanil were exposed to $H_2O_2$ for 24 h. We determined the cell viability of each group using an MTT assay. Hoechst staining and FACS analysis of Cos-7 cells were performed to observe the effect of remifentanil on apoptosis. Autophagy activation was determined by fluorescence microscopy, MDC staining, and AO staining. The expression of autophagy-related proteins was observed using western blotting. Results: Remifentanil pretreatment increased the viability of Cos-7 cells exposed to oxidative stress. Hoechst staining and FACS analysis revealed that oxidative stress-dependent apoptosis was suppressed by the pretreatment. Additionally, fluorescence microscopy showed that remifentanil pretreatment led to autophagy-induction in Cos-7 cells, and the expression of autophagy-related proteins was increased in the RPC+$H_2O_2$ group. Conclusions: The study showed that remifentanil pretreatment stimulated autophagy and increased viability in an oxidative stress model of Cos-7 cells. Therefore, we suggest that apoptosis was activated upon oxidative stress, and remifentanil preconditioning increased the survival rate of the cells by activating autophagy.

• Journal of The Korean Society of Grassland and Forage Science
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• v.38 no.4
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• pp.320-324
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• 2018

In this study, we examined effect of sperm penetration of oocytes after in vitro fertilization (IVF) with cauda epididymal spermatozoa in Hanwoo bull after feeding of timothy hay. One testicle with epididymides was castrated from one Hanwoo bull (14 months of age) and spermatozoa recovered from cauda epididymis and cryopreserved. As control, frozen Hanwoo semen was used. Matured cumulus oocyte complexes were co-incubated with frozen-thawed cauda epididymal spermatozoa for 12 or 18 hours. After IVF, presumptive zygotes were cultured in modified synthetic oviductal fluid. In experiment 1, we examined sperm penetration rate at 12 hours of IVF with epididymal sperm. Total penetration rate among cauda epididymis and control was similar(mean${\pm}$standard error, cauda epididymis and control vs. $49.7{\pm}11.3$ and $54.4{\pm}12.8%$). In experiment 2, cleavage and blastocyst developmental rate were evaluated at day 2 and day 8 after IVF for 18 hours. Cleavage rate among cauda epididymis and control was similar(cauda epididymis and control vs. $81.2{\pm}3.4$ and $82.7{\pm}2.5%$). However, blastocyst developmental rate of cauda epididymis group was significantly higher than that of control group(cauda epididymis and control vs. $24.4{\pm}1.6$ and $12.2{\pm}2.8%$, p<0.05). In conclusion, cauda epididymal spermatozoa in Hanwoo bull has high embryo developmental competence and can be used as an alternative to ejaculated frozen sperm in vitro.

• Journal of Animal Science and Technology
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• v.63 no.6
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• pp.1376-1385
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• 2021

Foot-and-mouth disease, one of the most contagious diseases in cloven-hoofed animals, causes significant economic losses. The pathogenesis of foot-and-mouth disease virus (FMDV) infection is known to differ with age of the animals. In this study, we aimed to reveal the difference in immunological response in the initial stage of FMDV infection between piglets and adult pigs. Peripheral blood mononuclear cells (PBMCs) were isolated from 3 piglets (8 weeks old) and 3 pigs (35 weeks old) that were not vaccinated against FMDV. O-type FMDV (2 × 10² median tissue culture infectious dose) was inoculated into porcine PBMCs and the cells were incubated at 37.0℃ under 5% CO₂ for various time periods (0, 1, 3, 6, 12, 24, and 48 h). The total RNA was obtained from the FMDV-inoculated PBMCs after each time point, and the virus titer was investigated in these RNA samples. Furthermore, dynamics of mRNA expression of the six tested cytokines (interferon [IFN]-α, IFN-γ, interleukin [IL]-6, IL-8, IL-10, and tumor necrosis factor [TNF]-α) in FMDV-inoculated porcine PBMCs were evaluated by time-series analysis to determine the differences, if any, based on the age of the pigs. The PBMCs of piglets contained the highest quantity of FMDV mRNA at 6 hours post-inoculation (hpi), and the PBMCs of pigs had the highest quantity of FMDV mRNA at 3 hpi. The mean cycle threshold-value in the PBMCs steadily decreased after the peak time point in the piglets and pigs (6 and 3 hpi, respectively). The dynamics of mRNA expression of all cytokines except TNF-α showed age-dependent differences in FMDV-inoculated PBMCs. The mRNA expression of most cytokines was more pronounced in the piglets than in the pigs, implying that the immune response against FMDV showed an age-dependent difference in pigs. In conclusion, within 48 hpi, the 8-week-old piglets responded more rapidly and were more sensitive to FMDV infection than the 35-week-old pigs, which could be associated with the difference in the pathogenesis of FMDV infection among the pigs. These results provide valuable insights into the mechanisms underlying the age-dependent differences in immune response in pigs against FMDV infection.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.8 no.2
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• pp.53-61
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• 2022

This study aimed to increase the targeting ability against PSMA in cell therapy using metabolic glycoengineering and biorthogonal chemistry and to visualize cell trafficking using PET imaging. Cellular membranes of THP-1 cells were decorated with azide(-N₃) using Ac₄ManNAz by metabolic glycoengineering. Engineered THP-1 cells were conjugated with DBCO-bearing fluorophore (ADIBO-Cy5.5) for 1 h at different concentrations and analyzed by confocal fluorescence microscopy and flow cytometry. For PSAM ligand conjugation to THP-1 cells, Ac₄ManNAz treated THP-1 cells were incubated with DBCO-PSMA ligand (ADIBO-GUL) at a final concentration with 100 µM for 1 h. To evaluate the effect on cell recognition, PSMA ligand conjugated THP-1 cells(as effectors) were co-cultured with PSMA positive 22RV1 (as target cells) at 3 : 1 a effector-to-target cell (E/T) ratio. The interaction between THP-1 and 22RV1 was monitored by confocal fluorescence microscopy. For preparing the radiolabeled THP-1, the cells were treated at the activity of ~ 740 kBq of [⁸⁹Zr]Zr(oxinate)₄/5 × 10⁶ cells. Radiolabeled cells were analyzed for determination of cell-associated radioactivity by gamma counting and viability using MTS assay. In the cytotoxicity assay, THP-1 cells did not have any cytotoxicity even when the Ac₄ManNAz concentration was 100 µM. In confocal microscopy and flow cytometry, THP-1 cells were efficiently labeled ADIBO-Cy5.5 in a dose-dependent manner, and the dose of 100 µM was the optimal concentration for the following experiments. The clusters of PSMA ligand-conjugated THP-1 cells and 22RV1 cells were identified, indicating cell-cell recognition over the cell surface between two types of cells. Cell radiolabeling efficiency was 54.5 ± 17.8%. THP-1 labeled with 0.09 ± 0.03 Bq/cell showed no significant cytotoxicity compared to unlabeled THP-1 up to 7 days. We successfully demonstrated that Ac₄ManNAz treated cells were efficiently conjugated with ADIBO-GUL for preparing the PSMA-targeting cells, and [⁸⁹Zr]Zr(oxinate)₄ could be used to label cells without toxicity. It suggested that PSMA-ligand conjugated cell therapy could be improved cell targeting and be monitored by PET imaging.

• Korean Journal of Clinical Laboratory Science
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• v.56 no.1
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• pp.52-58
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• 2024

Photodynamic therapy (PDT) activates intracellular oxygen using a photosensitizer activated by light of a specific wavelength and is a potential means of treating wound infections caused by antibiotic-resistant bacteria. Pseudomonas aeruginosa (P. aeruginosa) is typically non-pathogenic in healthy individuals but can induce severe illnesses like sepsis in the immunocompromised. Antibiotics have been conventionally used to treat P. aeruginosa infections, but increasing antibiotic resistance caused by drug misuse poses a growing challenge to the management of these infections. This study aimed to investigate the ability of PDT using photosensitizers (PhotoMed, Methyl pheophorbide A, or Radachlorin^®) and a diode laser to inhibit P. aeruginosa. Suspensions of P. aeruginosa and a photosensitizer were inoculated into Petri dishes and incubated for 30 minutes. Samples were then irradiated with the laser at 3 J/cm², and after incubation, colony areas were measured. P. aeruginosa killing rates were 79.65% for PhotoMed, 47.36% for Methyl pheophorbide A, and 40.91% for Radachlorin^®. This study shows that PDT using a diode laser and a photosensitizer constitutes an effective practical therapeutic approach for inhibiting P. aeruginosa.

• The Korean Journal of Nuclear Medicine Technology
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• v.14 no.2
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• pp.104-109
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• 2010

Purpose: $[^{18}F]$Fallypride plays an effective radiotracer for the study of dopamine $D_2/D_3$ receptor occupancy, neuropsychiatric disorders and aging in humans. This tracer has the potential for clinical use, but automated labeling efficiency showed low radiochemical yields about 5~20% with relatively long labelling time of fluorine-18. In present study, we describe an improved automatic synthesis of [$^{18}F$]Fallypride using different base concentration for routine clinical use. Materials and Methods: Fully automated synthetic process of [$^{18}F$]Fallypride was perform using the TracerLab $FX_{FN}$ synthesizer under various labeling conditions and tosyl-fallypride was used as a precursor. [$^{18}F$]Fluoride was extracted with various concentration of $K_{2.2.2.}/K_2CO_3$ from $^{18}O$-enriched water trapped on the ion exchange cartridge. After azeotropic drying, the labeling reaction proceeded in $CH_3CN$ at $100^{\circ}C$ for 10 or 30 min. The reaction mixture was purified by reverse phase HPLC and collected organic solution was exchanged by tc-18 Sep-Pak for the clinically available solution. Results: The optimal labeling condition of [$^{18}F$]Fallypride in the automatic production was that 2 mg of tosyl-fallypride in acetonitrile (1 mL) was incubated at $100^{\circ}C$ for 10 min with $K_{2.2.2.}/K_2CO_3$ (11/0.8 mg). [$^{18}F$]Fallypride was obtained with high radiochemical yield about $66{\pm}1.4%$ (decay-corrected, n=28) within $51{\pm}1.2$ min including HPLC purification and solid-phase purification for the final formulation. Conclusion: [$^{18}F$]Fallypride was prepared with a significantly improved radiochemical yield with high specific activity and shorten synthetic time. In addition, this automated procedure provides the high reproducibility with no synthesis failures (n=28).

• Journal of Animal Science and Technology
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• v.55 no.1
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• pp.25-32
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• 2013

The purpose of this study was to investigate the effects of mustard, which contains allyl isothiocyanate, on ruminal fermentation and methane emission in vitro. To this end, diluted ruminal fluid(30ml) was incubated anaerobically at $39^{\circ}C$ or 6, 12, and 24 h with or without seeds or powdered mustard. Either mustard seed or powdered mustard was weighed and serially (0, 3.33, 5.00, 6.67, and 8.33 g/L) mixed with ruminal fluid. Ammonia-N was increased (P < 0.05) by mustard treatment in a dose dependent manner. Regardless of concentration or form, mustard increased (P < 0.05) total VFA content but decreased (P < 0.01) pH compared to control group. Molar proportion of acetate (A) was decreased (P < 0.05) whereas propionate (P) was increased (P < 0.05) by mustard treatment, thereby A:P ratio was decreased (P < 0.05) compared to control group. Total gas production was increased (P < 0.01) in a linear manner by mustard treatment compared to control group. There was no effect of mustard powder, except 8.33 g/L level at 6 h, on methane emission. However, at 24 h, methane emission was reduced (P < 0.05) by 4.77% and 11.54% with 6.67 g/L and 8.33 g/L of mustard seeds supplementation, respectively. Altogether, these results suggest that mustard seeds containing allyl isothiocyanate may reduce methane production without disturbing ruminal fermentation.

• Journal of Animal Science and Technology
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• v.54 no.5
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• pp.369-373
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• 2012

AMP-activated protein kinase (AMPK) maintains energy homeostasis in skeletal muscle. Nonetheless, its functional role on protein synthesis with different nutrient availability has not been elucidated. Therefore, the purpose of this study is to examine the effect of AMPK activity on protein content in C2C12 myotubes incubated with low (5 mM; LG) or high (25 mM; HG) glucose media. LG stimulated (p<0.05) AMPK and acetyl CoA carboxylase (ACC) activity compare to those in HG group. Total protein content was higher in myotubes cultured with HG than in those cultured with LG and further increased by AICAR (5-amino-1-${\beta}$-D-ribofuranosyl-imidazole-4-carboxamide). Myotubes cultured with HG showed 7.5% lower UbFL (Ubiquitin Firefly Luciferase)-to-SV40 (Simian virus40) ratio compared to those in LG. Glucose level did not change the phosphorylation level of mammalian target of rapamycin (mTOR). Interestingly, administration of AICAR significantly increased phosphorylation level of mTOR in myotubes cultured with LG but not in those with HG. Overall, this data indicate that AMPK activity and protein turnover are finely regulated in response to different glucose concentration.