• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.257-277
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• 2000

New techniques for regenerating the destructed periodontal tissue have been studied for many years. Current acceptable methods of promoting periodontal regeneration are basis of removal of diseased soft tissue, root treatment, guided tissue regeneration, graft materials, and biological mediators. Platelet Rich Plasma has been reported as a biological mediator which regulates activities of wound healing progress including cell proliferation, migration, and metabolism. The purpose of this study is to evaluate the effects of using the Platelet Rich Plasma as a regeneration promoting agent for furcation involvement defect. Five adult beagle dogs were used in this experiment. The dogs were anesthetized with Ketamin HCl(0.1 ml/kg, IV)and Xylazine hydrochloride($Rompun^{(R)}$, Bayer, 0.1 ml/kg, IM) and conventional periodontal prophylaxis were performed with ultrasonic scaler and hand instruments. With intrasulcular and crestal incision, mucoperiosteal flap was elevated. Following decortication with 1/2 high speed round bur, degree II furcation defect was made on mandibular third(P3), forth(P4) and fifth(P5) premolar, and stopping was inserted. After 4 weeks, stopping was removed, and bone graft was performed. Ca-P was grafted in P3(experimental group I), Combination of Ca-P and plasma rich platelet were grafted in P4(experimental group II), and P5 was remained at control group.Systemic antibiotics(gentamicin sulfate)and anlgesics(phenyl butazone) were administrated intramuscular for 2 weeks after surgery. Irrigation with 0.1% Chlorhexidine Gluconate around operate sites was performed during the whole experimental period except one day immediate after surgery. Soft diets were fed through the whole experiment period. After 4, 8 weeks, the animals were sacrificed by perfusion technique. Tissue block was excised including the tooth and prepared for light microscope with Gomori's trichrome staining. At 4 weeks after surgery, there were rapid osteogenesis phenomenon on the defected area of the Platelet Rich Plasma plus Ca-P BBP group and early trabeculation pattern was made with new osteoid tissue produced by activated osteoblast. Bone formation was almost completed to the fornix of furcation by 8 weeks after surgery. In conclusion, Platelet Rich Plasma can promote rapid osteogenesis during healing of periodontalregeneration.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.2
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• pp.172-178
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• 2002

The present study examined whether treatment of in vitro matured pig oocytes with calcium ionophore (A23187) could prevent polyspermic penetration in vitro. When oocytes cultured for maturation for 33, 36 or 44 h were subsequently treated with $50{\mu}M$ A23187 in medium with fetal calf serum (FCS) for 1, 2 and 3 h and then cultured for 12 h without spermatozoa, virtually no activation occurred. In the absence of FCS, however, 31-42, 45-49 and 56-64% of oocytes were activated, respectively. When oocytes treated with $50 {\mu}M$ A23187 in medium with FCS for 3 h were inseminated in vitro, the penetration rates (14-57%) were lower (p<0.01) with a higher (p<0.01) incidence (35-67%) of monospermy compared with untreated oocytes (69-80% penetration and 15-17% monospermy). However, sperm penetration was completely blocked in all oocytes treated with A23187 in the absence of FCS. When oocytes matured for 33 h were treated with different concentrations of A23187 for 3 h and inseminated in vitro, the penetration rate did not change but there was an increased incidence (p<0.05) of monospermy at $10-20{\mu}M$ and $2.5-5{\mu}M$ A23187 in the presence and absence of FCS, respectively, compared with at $0{\mu}M$ A23187. With these lower concentrations of A23187, treatment of oocytes for at least 60 and 30 min in the presence and absence of FCS, respectively, was required to increase the incidence of monospermy without reducing penetration rate. These results indicate that a high concentration ($50{\mu}M$) of A23187 in medium without FCS, but not in medium with FCS, stimulated in vitro matured pig oocytes to induce parthenogenetic activation and a complete block to sperm penetration in vitro. However, treatment of oocytes with lower concentrations of A23187 ( $10-20{\mu}M$ and $2.5-5{\mu}M$) both in the presence and absence of FCS maintained sperm penetration in vitro and increased the incidence of monospermy.

• Radiation Oncology Journal
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• v.17 no.2
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• pp.113-119
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• 1999

Purpose : To evaluate possible acute toxicity and early response of concurrent radiation therapy and low dose daily cisplatin as a radiosensitizer in patients with locally advanced uterine cervical carcinomas. Materials and Method : From December 1996 to January 1999, 38 previously untreated Patients with locally advanced squamous cell carcinoma of the uterine cervix (from stage IIB to stage IIIB) were treated at Inha University Hospital. All patients underwent standard pretreatment staging Procedures after the initial evaluation by gynecologists and radiation oncologists. Sixteen Patients with huge cervical mass (>4 cm) were submitted to the group treated with concurrent radiation therapy and low dose daily cisplatin while the remainder was treated with radiation therapy alone. Radiation therapy consisted of 4500 cGy external beam irradiation to whole pelvis (midline block after 3000 cGy), 900$\~$1000 cGy boost to involved parametrium, and high dose-rate intracavitary brachytherapy (a total dose of 3000$\~$3500 cGy/500 cGy per fraction to point A, twice per week). In the group treated with low dose cisplatin concurrently, 10 mg of daily intravenous cisplatin was given from the 1st day of radiation therapy to the 20th day of radiation therapy. Acute toxicity was measured according to expanded common toxicity criteria of the NCI (C) Clinical Trials. Early response data were analyzed at minimum 4 weeks' follow-up after completion of the treatment protocol. Results: Hematolgic toxici쇼 was more prominent in patients treated with radiation therapy and cisplatin. Six of 16 patients (37.5$\~$) treated with radiation therapy and cisplatin and one of 22 patients (4.5$\~$) treated with radiation therapy alone experienced grade 3 leukopenia. In Fisher's exact test, there was statistically significant difference between two groups regarding leukopenia (P=0.030). There was no apparent difference in the frequency of gastrointestinal and genitourinary toxicity between two groups (P=0.066). Three of 16 patients (18.7$\~$) treated with radiation therapy and cisplatin and two of 22 patients (9.1$\~$) treated with radiation therapy alone experienced more than 5 kg weight loss during the treatment. There was no statistically significant difference on weight loss between two groups (P=0.63). Two patients on each group were not evaluable for the early response because of incomplete treatment. The complete response rate at four weeks' follow-up was 80$\~$(16/20) for the radiation therapy alone group and 78$\~$ (11/14) for the radiation therapy and cisplatin group. There was no statistically significant difference in early response between two treatment groups (P=0.126). Conclusion : This study led to the conclusion that the hematologic toxicity from the treatment with concurrent radiation therapy and low dose daily cisplatin seems to be more prominent than that from the treatment of radiation therapy alone. There was no grade 4 hematologic toxicity or mortality in both groups. The hematologic toxicity in both treatment groups seems to be well managable modically. Since the risk factors were not balanced between two treatment groups, the direct comparison of early response of both groups was not possible. However, preliminary results regarding early response for patients with bulky cervical tumor mass treated with radiation therapy and low dose daily cisplatin was encouraging. Longer follow-up is necessary to evaluate the survival data. A phase III study is needed to evaluate the efficacy of concurrent daily low dose cisplatin with radiation therapy in bulky cervical cancer.

• The Korean Journal of Physiology
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• v.18 no.1
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• pp.19-35
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• 1984

Since the first report of Drury and $Szent-Gy{\ddot{o}}rgyi$ in 1929, the inhibitory influences of adenosine on the heart have repeatedly been described by many investigators. These studies have shown that adenosine and adenine nucleotides have overall depressant effects, similar to those of acetylcholine. Heart beats become slow and weak. It is also well known that adenosine is a potent endogenous coronary vasodilator. Many investigations on the working mechanisms of adenosine have been focused mainly on the effects of the coronary blood flow. However, the cellular mechanisms underlying the inhibitory action of adenosine on sinus node are not well understood yet. Thus, this study was undertaken to examine the behavior of rabbit SA node under influence of adenosine. In these series of experiments three kinds of preparations were used: whole atrial pair, left atrial strip, and isolated SA node preparations. The electrical activity of SA node was recorded with conventional glass microelectrodes 30 to 50 $M{\Omega}$. The preparations were superfused with bicarbonate-buffered Tyrode solution of pH 7.35 and aerated with a gas mixture of $3%\;CO_2-97%\;O_2$ at $35^{\circ}C$. In whole atrial pair, adenosine suppressed sinoatrial rhythm in a dose-dependent manner. Effect of adenosine on atrial rate appeared at the concentration of $10^{-5}M$ and was enhanced in parallel with the increase in adenosine concentration. Inhibitory action of adenosine on pacemaker activity was more prominent in the preparation pretreated with norepinephrine, which can steepen the slope of pacemaker potential by increasing permeability of $Ca^{+2}$. Calcium ions in perfusate slowly produced a marked change in sinoatrial rhythm. Elevation of the calcium concentration from 0.3 to 8 mM increased the atrial rate from 132 to 174 beats/min, but over 10 mM $Ca^{+2}$ decreased. The inhibitory effect of adenosine on sinoatrial rhythm developed very rapidly. Atrial rate was recovered promptly from the adenosine-induced suppression by the addition of norepinephrine, but extra $Ca^{+2}$ was less suitable to restore the suppression of atrial rate. Adenosine suppressed also atrial contractility in the same dosage range that restricted pacemaker activity, even in the reserpinized preparation. In isolated SA node preparation, spontaneous firing rate of SA node at $35^{\circ}C$(mean{\pm}SEM, n=16) was $154{\pm}3.3\;beats/min. The parameters of action potentials were: maximum diastolic potential(MDP), $-73{\pm}1.7\;mV: overshoot(OS), $9{\pm}1.4\;mV: slope of pacemaker potential(SPP), $94{\pm}3.0\;mV/sec. Adenosine suppressed the firing rate of SA node in a dose-dependent manner. This inhibitory effect appeared at the concentration of $10^{-6}M$ and was in parallel with the increase in adenosine concentration. Changes in action potential by adenosine were dose-dependent increase of MDP and decrease of SPP until $10^{-4}M$. Above this concentration, however, the amplitude of action potential decreased markedly due to the simultaneous decrease of both MDP and OS. All these effects of adenosine were not affected by pretreatment of atropine and propranolol. Lowering extra $Ca^{2+}$ irom 2 mM to 0.3 mM resulted in a marked decrease of OS and SPP, but almost no change of MDP. However, increase of perfusate $Ca^{2+}$ from 2 mM to 6 or 8 mM produced a prominent decrease of MDP and a slight increase of OS and SPP. Dipyridamole(DPM), which is known to block the adenosine transport across the cell membrane, definately potentiated the action of adenosine. The results of this experiment suggest that adenosine suppressed pacemaker activity and atrial contractility simultaneously and directly, by decreasing $Ca^{2+}-permeability$ of nodal and atrial cell membranes.

• The Korean Journal of Pain
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• v.13 no.2
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• pp.149-155
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• 2000

Background: Pulsed radiofrequency (RF) lesioning is a painless procedure and causes no neurodestruction and neuritis-like reaction are common following conventional RF lesioning. There is little data about the effect of pulsed RF especially with regard to its suitability for the treatment of acute pain. The possibility of a placebo effect cannot be ruled out because a double-blind study was not performed in previous studies. There is also no neuropathologic study about pulsed RF. Methods: The rats were anesthetized with sodium pentobarbital (40 mg/kg, i.p.; supplemented as necessary). The common sciatic nerve was exposed by blunt dissection through biceps femoris. Pulsed RF was administered to the common sciatic nerve using a 30 ms/s pulse with for 120 seconds. The temperature reached was no more than $42^{\circ}C$. Analgesia was determined using hot-plate assay shortly and, 3 days and 1 week before, and 2 weeks after operation. Lesions were examined with LM (light microscope) and EM (electron microscope) 2 weeks later. Results: There were no differences in response latencies between the control and experimental group. There were many vacuoles with hyaline bodies in the Schwann cell cytoplasm rather than axon in LM and larger electron dense bodies. No changes were found in the axon or unmyelinated fibers. Only small changes were found in the sheaths of myelinated fibers and Schwann cells. Conclusions: We therefore do think that any analgesic effect of pulsed RF is not a result of block of neural conduction. But rather than it can be attributed to others factors. It was also ineffective as a treatment for acute pain such as that caused by the hot-plate test.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.2
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• pp.204-213
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• 2008

Four ruminally cannulated Holstein cows averaging 43 days in milk (DIM) were used in a $4{\times}4$ Latin square to determine the effect of feeding extruded soybean, ground canola seed and whole cottonseed on ruminal fermentation and milk fatty acid profile. One hundred and twenty lactating Holstein cows, 58 (${\pm}31$) DIM, were assigned to four treatments in a completely randomized block design to study the effects of the three types of oilseeds on production parameters and milk fatty acid profile. The four diets were a control diet (CON) and three diets in which 10% extruded soybean (ESB), 5% ground canola seed (GCS) and 10% whole cottonseed (WCS) were included, respectively. Diets consisted of concentrate mix, corn silage and Chinese wild rye and were balanced to similar concentrations of CP, NDF and ADF. Ruminal fermentation results showed that ruminal fermentation parameters, dry matter intake and milk yield were not significantly affected by treatments. However, compared with the control, feeding cows with the three oilseed diets reduced C14:0 and C16:0 and elevated C18:0 and C18:1 concentrations in milk, and feeding ESB increased C18:2 and cis9, trans11 conjugated linoleic acid (CLA). Production results showed that feeding ESB tended to increase actual milk yield (30.85 kg/d vs. 29.29 kg/d) and significantly decreased milk fat percentage (3.53% vs. 4.06%) compared with CON. Milk protein (3.41%) and solid non-fat (13.27%) from cows fed WCS were significantly higher than from cows fed CON (3.24% and 12.63%, respectively). Milk urea N concentrations from cows fed the ESB (164.12 mg/L) and GCS (169.91 mg/L) were higher than cows fed CON (132.31 mg/L). However, intake of DM, 4% fat corrected milk, energy corrected milk, milk fat and protein yields, milk lactose percentage and yield, somatic cell count and body condition score were not affected by different treatments. The proportion of medium-chain fatty acid with 14 to 16 C units in milk was greatly decreased in cows fed ESB, GCS and WCS. Feeding ESB increased the concentration in milk of C18:1, C18:2, C18:3 and cis9, trans11-CLA content by 16.67%, 37.36%, 95.24%, 72.22%, respectively, feeding GCS improved C18:0 and C18:1 by 17.41% and 33.28%, respectively, and feeding WCS increased C18:0 by 31.01% compared with feeding CON. Both ruminal fermentation and production trial results indicated that supplementation of extruded soybean, ground canola seed and whole cottonseed could elevate the desirable poly- and monounsaturated fatty acid and decrease the medium chain fatty acid and saturated fatty acid content of milk fat without negative effects on ruminal fermentation and lactation performance.

• Reproductive and Developmental Biology
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• v.32 no.3
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• pp.205-209
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• 2008

This study was carried out to develop knock-in vector for EGFP (enhanced green fluorescent protein) expression in porcine $\beta$-casein locus. For construction of knock-in vector using porcine $\beta$-casein gene, we cloned the $\beta$-casein genome DNA from porcine fetal fibroblast cells, EGFP and SV40 polyA signal using PCR. The knock-in vectors consisted of a 5-kb fragment as the 5' recombination arm and a 2.7-kb fragment as the 3' recombination arm. We used the neomycin resistance gene ($neo^{r}$) as a positive selectable marker and the diphtheria toxin A (DT-A) gene as a negative selectable marker. To demonstrate EGFP expression from knock-in vector, we are transfected knock-in vector that has EGFP gene in murine mammary epithelial cell line HC11 cells with pSV2 neo plasmid. The EGFP expression was detected in HC11 cells transfected knock-in vector. This result demonstrates that this knock-in vector may be used for the development of knock-in transgenic pig.

• Korean Journal of Organic Agriculture
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• v.26 no.1
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• pp.151-161
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• 2018

This study was designed to investigate the effects of multi-enzyme on diarrhea and immune responses of weaned pigs. A total 36 weaned pigs ($5.92{\pm}0.48kg\;BW$; 28 d old) were randomly allotted to 2 dietary treatments (3 pigs/pen, 6 replicates/treatment) in a randomized complete block design. The dietary treatments were a typical diet based on corn and soybean meal (CON) and CON with 0.1% multienzyme (Multi; mixture of ${\beta}-mannanase$, xylanase, ${\alpha}-amylase$, protease, ${\beta}-glucanase$, and pectinase). Pigs were fed their respective diets for 6 wk. Frequency of diarrhea, levels of packed cell volume (PCV), white blood cells (WBC), immunoglobulins, cortisol, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), transforming growth $factor-{\beta}$ ($TGF-{\beta}$), and C-reactive protein (CRP) were measured. Multi group tended to decrease (p<0.1) diarrhea frequency than CON group during 2 wk after weaning. Lower values of PCV on d 3 (p<0.05) and d 7 (p<0.1) were found in Multi group compared with CON group. There were no significant differences on WBC number and immunoglobulin (Ig) M and A between Multi and CON groups. However, Multi group tended to increase (p<0.1) Ig G on d 7 than CON group. Moreover, Multi group showed modulated immune responses, indicated by decreased levels of cortisol (p<0.05) on d 7 and 14, $TNF-{\alpha}$ on d 3 (p<0.05) and d 7 (p<0.10), $TGF-{\beta}$ on d 2 (p<0.05) and d 7 (p<0.10), and CRP (p<0.10) on d 3 and 7 after weaning compared with CON group. Consequently, inclusion of multi-enzyme in diets for weaned pigs improved gut health and modulated immune responses of weaned pigs.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.4
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• pp.271-279
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• 2019

The lamina II, also called the substantia gelatinosa (SG), of the trigeminal subnucleus caudalis (Vc), is thought to play an essential role in the control of orofacial nociception. Glycine and serotonin (5-hydroxytryptamine, 5-HT) are the important neurotransmitters that have the individual parts on the modulation of nociceptive transmission. However, the electrophysiological effects of 5-HT on the glycine receptors on SG neurons of the Vc have not been well studied yet. For this reason, we applied the whole-cell patch clamp technique to explore the interaction of intracellular signal transduction between 5-HT and the glycine receptors on SG neurons of the Vc in mice. In nine of 13 neurons tested (69.2%), pretreatment with 5-HT potentiated glycine-induced current ($I_{Gly}$). Firstly, we examined with a $5-HT_1$ receptor agonist (8-OH-DPAT, $5-HT_{1/7}$ agonist, co-applied with SB-269970, $5-HT_7$ antagonist) and antagonist (WAY-100635), but $5-HT_1$ receptor agonist did not increase $I_{Gly}$ and in the presence of $5-HT_1$ antagonist, the potentiation of 5-HT on $I_{Gly}$ still happened. However, an agonist (${\alpha}$-methyl-5-HT) and antagonist (ketanserin) of the $5-HT_2$ receptor mimicked and inhibited the enhancing effect of 5-HT on $I_{Gly}$ in the SG neurons, respectively. We also verified the role of the $5-HT_7$ receptor by using a $5-HT_7$ antagonist (SB-269970) but it also did not block the enhancement of 5-HT on $I_{Gly}$. Our study demonstrated that 5-HT facilitated $I_{Gly}$ in the SG neurons of the Vc through the $5-HT_2$ receptor. The interaction between 5-HT and glycine appears to have a significant role in modulating the transmission of the nociceptive pathway.

• Journal of Microbiology and Biotechnology
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• v.31 no.7
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• pp.942-948
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• 2021

Canine influenza virus (CIV) induces acute respiratory disease in dogs. In this study, we aimed to determine the signaling pathways leading to the induction of IFN-β in a canine respiratory epithelial cell line (KU-CBE) infected with the H3N2 subtype of CIV. Small interfering RNAs (siRNAs) specific to pattern recognition receptors (PRRs) and transcription factors were used to block the IFN-β induction signals in H3N2 CIV-infected KU-CBE cells. Among the PRRs, only the TLR3 and RIG-I expression levels significantly (p < 0.001) increased in CIV-infected cells. Following transfection with siRNA specific to TLR3 (siTLR3) or RIG-I (siRIG-I), the mRNA expression levels of IFN-β significantly (p < 0.001) decreased, and the protein expression of IFN-β also decreased in infected cells. In addition, co-transfection with both siTLR3 and siRIG-I significantly reduced IRF3 (p < 0.001) and IFN-β (p < 0.001) mRNA levels. Moreover, the protein concentration of IFN-β was significantly (p < 0.01) lower in cells co-transfected with both siTLR3 and siRIG-I than in cells transfected with either siTLR3 or siRIG-I alone. Also, the antiviral protein MX1 was only expressed in KU-CBE cells infected with CIV or treated with IFN-β or IFN-α. Thus, we speculate that IFN-β further induces MX1 expression, which might suppress CIV replication. Taken together, these data indicate that TLR3 and RIG-I synergistically induce IFN-β expression via the activation of IRF3, and the produced IFN-β further induces the production of MX1, which would suppress CIV replication in CIV-infected cells.