• The Journal of Advanced Prosthodontics
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• v.3 no.4
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• pp.186-189
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• 2011

PURPOSE. Porcelain fused to metal (PFM) crowns provide the best treatment option for teeth that have a large or defective restoration. More than 20% of teeth with PFM crowns or bridges require non-surgical root canal treatment (NSRCT). This may be due to the effect of restorative procedures and the possible leakage of bacteria and or their by-products, which leads to the demise of the tooth pulp. Thus, this study was planned to compare the ability of the restorative materials to seal perforated PFM specimens. MATERIALS AND METHODS. The study evaluates the ability of amalgam, composite or compomer restorative materials to close perforated PFM specimen's in-vitro. Ninety PFM specimens were constructed using Ni-Cr alloys and feldspathic porcelain, and then they were divided into 3 groups: amalgam (A), composite + Exite adhesive bond (B) and compomer + Syntac adhesive bond (C). All the PFM samples were embedded in an acrylic block to provide complete sealing of the hole from the bottom side. After the aging period, each group was further divided into 3 equal subgroups according to the thermocycling period (one week for 70 cycles, one month for 300 cycles and three months for 900 cycles). Each subgroup was put into containers containing dye (Pelikan INK), one maintained at $5^{\circ}C$ and the other at $55^{\circ}C$, each cycle for 30 sec time. The data obtained was analyzed by SPSS, 2006 using one way ANOVA test and student t-test and significant difference level at (P<.01). RESULTS. The depth of dye penetration was measured at the interfaces of PFM and filling materials using Co-ordinate Vernier Microscope. The lowest levels of the dye penetration for the three groups, as well as subgroups were during the first week. The values of dye leakage had significantly increased by time intervals in subgroups A and C. CONCLUSION. It was seen that amalgam showed higher leakage than composite while compomer showed the lowest level of leakage.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.23-29
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• 2000

Objective: Zona pellucida (ZP) has been thought to be the barrier of egg to sperm penetration before and after fertilization. The phenomenon of ZP hardening has been considered as a post-fertilization event until now, and it is generally accepted that it is caused by the secretory products of cortical granules released during the cortical reaction. Hardening of ZP could occur "spontaneously" in mammalian oocytes in standard culture conditions, and that it is probably not a consequence of cortical reaction. The purpose of our study was to investigate the effect of human amniotic fluid (HAF) on nuclear maturation (NM) and fertilization ability of mouse immature oocytes. Methods: HAF was obtained from patients undergoing amniocentesis at $16{\sim}20$ weeks of gestation. HAF from five to ten patients was centrifuged and the supernatants was pooled. Cumulusenclosed mouse immature oocytes were incubated in the medium containing HAF, and examined to confirm NM and fertilization. Female ICR mice (about 3 weeks old) were stimulated with 7.5 IU PMSG. Immature oocytes were isolated at $48{\sim}52$ hrs post PMSG injection and cultured in TCM-199 supplemented with 20% HAF for 18 hrs. FBS was used as a control for the examination. Matured oocytes (MII) were fertilized with sperms collected from the epididymis of male mice (over 10 weeks old). Fertilization was in conducted T6 medium containing 15 mg/ml BSA, and confirmed at 6 hrs post-insemination. Fertilization rate was assessed in zona-intact or zona-free oocytes (denuded by trypsin). Evaluation of NM and fertilization was carried out by rapid staining method. ZP hardening was evaluated by incubating cumulus cell-free mature oocytes in 0.001% chymotrypsin at $37^{\circ}C$ for 10 min. Results: There was no significant difference between the effects of HAF (86.6%) and FBS (87.7%) supplements on NM of immature oocytes. When maturation medium was supplemented with HAF, total fertilization rates (7%) were significantly lower (p<0.01) than that of FBS (85.1%). In HAF group, fertilization rate was increased (p<0.01) in zona-free oocytes (7% versus 100%). The resistance of mouse oocyte ZP to digestion by chymotrypsin after maturation in vitro was significantly higher (p<0.01) in HAF group (86.7%) than in FBS (6.7%). To culture oocytes in FBS were very effective in preventing ZP hardening. However cultured oocytes in HAF showed high rate of ZP hardening (p<0.01). Conclusions: These results suggest that HAF can be used as a supplement for the NM of mouse immature oocytes in vitro. However, HAF induces spontaneous hardening of ZP of mouse immaure oocytes during maturation in vitro.

• Journal of Pharmaceutical Investigation
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• v.36 no.5
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• pp.321-329
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• 2006

Albuterol, a selective ${\beta}_2$-adrenergic receptor stimulant, has been introduced as a potent bronchodilator for patients with bronchial asthma, chronic obstructive bronchial disease, chronic bronchitis and pulmonary emphysema. The percutaneous permeation of albuterol sulfate was investigated in hairless mouse skin in vitro with and without pretreatment with enhancers. The enhancing effects of ethanol and various penetration enhancers such as terpenes, non-ionic surfactants, pyrrolidones, and fatty acids on the permeation of albuterol sulfate were evaluated using Franz diffusion cells. Among terpenes studied, 1,8-cineole was the most effective enhancer, which increased the permeability of albuterol sulfate approximately 33-fold compared with the control without enhancer pretrement, followed by d-limonene with enhancement ratio of 21.79. 2-Pyrrolidone-5-carboxylic acid increased the permeability of albuterol sulfate approximately 5.5-fold compared with the control. Other pyrrolidones tested showed only slight permeability enhancing effect with enhancement ratio less than 2.8. Nonionic surfactants showed moderate enhancing effects. Lauric acid increased the permeability of albuterol sulfate approximately 30-fold with decreasing the lag time from 2.85 to 0.64 hr. Oleic acid and linoleic acid showed enhancement ratio of 24.55 and 22.91, respectively. These findings would allow a more rational approach for designing formulations for the transdermal delivery of albuterol sulfate and similar drugs.

• Journal of Veterinary Clinics
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• v.35 no.2
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• pp.39-45
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• 2018

We evaluated the effects of green tea extract (GTE) supplementation at different dilution steps on boar sperm freezing and in vitro fertilization. Sperm intracellular hydrogen peroxide ($H_2O_2$), motility, viability, acrosome integrity and morphology were determined. In addition, sperm IVF parameters (penetration and monospermy) and glutathione (GSH) levels of presumptive zygotes (PZs) were evaluated. Semen was diluted in lactose egg yolk (LEY) and cooled at $5^{\circ}C$ for 3 h (first dilution step) and then diluted in LEY with 9% glycerol and maintained at $5^{\circ}C$ for 30 min (second dilution step). Four experimental groups were compared: first and second dilution steps without GTE (control), first dilution step with GTE (Step 1), second dilution step with GTE (Step 2) and first and second dilution step with GTE (Step 1+2). The spermatozoa were frozen in nitrogen vapor. Higher sperm motility, viability and acrosome integrity after thawing were observed in Step 1, Step 2 and Step 1+2 groups compared with the control (P < 0.05). Lower $H_2O_2$ level was observed in Step 1+2 compared with control and Step 1 (P < 0.05). For IVF, matured oocytes were co-cultured with spermatozoa frozen according to the experimental groups. GSH levels of PZs were significantly higher in Step 2 and Step 1+2 than in control and Step 1 (P < 0.05) without a significant difference in IVF parameters. In conclusion, supplementation with GTE in both first and second dilution steps during the freezing process resulted in better boar sperm cryopreservation and might be beneficial for further embryo development.

• Journal of Embryo Transfer
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• v.32 no.4
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• pp.337-342
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• 2017

The present study was aimed to determine the effects of green tea extract (GTE) and beta-mercaptoethanol (${\beta}-ME$) supplementation in boar sperm freezing extender on in vitro fertilization (IVF) and reactive oxygen species (ROS) and glutathione (GSH) levels of presumptive zygotes (PZs). Experimental groups were allocated into lactose egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/l in LEY) and ${\beta}-ME$ ($50{\mu}M$ in LEY). In freezing, spermatozoa extended with LEY were cooled to $5^{\circ}C$ for 3 h and then kept at $5^{\circ}C$ for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM. The final sperm concentration was $1{\times}10^8/ml$. Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. For IVF, oocytes were matured in NCSU-23 medium and co-cultured with spermatozoa following thawing at $37^{\circ}C$ for 25 sec. At 12 h following IVF, IVF parameters (sperm penetration and monospermy) were evaluated. In addition, GSH and ROS levels of PZs were determined by Cell Tracker Blue CMF2HC and DCHFDA, respectively. IVF parameters did not show any significant difference among the experimental groups. GSH and ROS levels of PZs were not significantly different between groups. In conclusion, antioxidant supplementation in boar sperm freezing could not influence IVF parameters, ROS and GSH levels of PZs.

• Journal of Embryo Transfer
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• v.17 no.3
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• pp.239-249
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• 2002

Embryos derived from pig oocytes matured in mSOF are able to develop to blastocysts after IVF. Experiment 1 evaluated the effects of two maturation media (TCM-199 vs mSOF) on maturation rate, fertilization parameters, including penetration, polyspermy, male pronuclear formation, and the mean number of sperm penetrated per oocyte. Experiment 2 and Experiments 3 examined the effects of two maturation media on zona pellucida solubility and cortical granule distribution by transmissible electron microscopy, respectively. Experiment 4 assessed the effects of two maturation media on the in vitro embryo cleavage rate and development to blastocyst. Lastly, experiment 5 examined the cell number of blastocyst. An effect of media (P<0.05) was detected for mSOF on the mean number of sperm per oocyte. In TCM group, zona digestion time (196.5$\pm$15.5 vs 131.6$\pm$20.1 before IVF, 397.5$\pm$30.3s vs 185.3$\pm$16.4s after IVF, p<0.05) was higher in TCM-199 group. No significant effects of media was observed on cortical granule distribution between two groups by TEM. An effect (P<0.05) was observed on embryo development to blastocyst (16% vs 8%) but not on cleavage rates. No significant effects of media was observed on total cell number of blastocyst. We found that the high mean number of sperm penetrated per oocyte and the weaker zona pellucida on the basis of the digestion time was shown in pig oocytes matured in mSOF, however, porcine oocyte maturation with supplemented synthetic oviduct fluid medium (mSOF) resulted in blastocyst cell numbers comparable to those observed with Tissue Culture Medium 199.

• Journal of Embryo Transfer
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• v.13 no.2
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• pp.139-145
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• 1998

In the present study, effects of interleukin-2 (IL-2), a differentiator and proliferator of T-cells, on nuclear maturation and sperm penetration of bovine oocytes was examined in a serum-free or serum-containing medium. Basic medium was used TCM-199 supplemented with 2.2g / ι sodium bicarbonate, 100 i.u. /rnl penicillin. 100$\mu$g /ml streptomycin, 0.25$\mu$g/ml Fungizone, this medium treated with FCS and IL-2. In experiment 1, we examined the effect of the addition of 0, 1, 5, 10 or 15nM /ml IL-2 to tissue culture medium (TCM-199) on nuclear maturation of oocytes Development of oocytes to the Metaphase II (M II) stage (%) was significantly (P<0.05) higher at 1, 5,10 and 15 nM /ml IL-2(54.2, 73.5, 80.0 and 69.6%, respectively) than at 0 nM /ml IL-2(35.7%). In experiment 2, we examined the effect of the addition of l0nM /ml IL-2 or 5% FCS in oocyte maturation. Nuclear maturation rates were significantly(P<0.05) higher l0nM /ml IL-2(80%) than non-treatment(35.7%) and 5% FCS(63.6%) treatment. On the other hand, there were no significant difference in the proportion of oocytes developed to the 2-cell stage after addition of IL-2 and/or FCS. These results suggest that IL-2 supports nuclear maturation of bovine immature oocytes in vitro. Serum-free maturation system using IL-2 might be useful for evaluation of various factors on oocyte maturation.

• Journal of the Korean Applied Science and Technology
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• v.31 no.2
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• pp.167-175
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• 2014

The influences of fluorescence, scattering, and flocculation in turbid material were interpreted for the scattered fluorescence intensity and wavelength, it has been studied the molecular properties by the spectroscopy of laser induced fluorescence(LIF). The effects of optical properties in scattering media have been found by the optical parameters(${\mu}_s$, ${\mu}_a$, ${\mu}_t$). Flocculation is an important step in many solid-liquid separation processes and is widely used in Photodynamic therapy. The interactions of several colloid particles can come into play which have major effect on the flocculation and LIF process. We measured scattering and fluorescence spectra of the sample for in vitro as function of concentration from lase source to detector. The value of scattering coefficient ${\mu}_s$ is large by means of the increasing particles of scatterer. Therefore, Phorphyrin A is larger than Phorphyrin C in scattering intensity ${\mu}_s$, but Phorphyrin A is smaller than Phorphyrin C in penetration depth ${\delta}$.

• Korean Journal of Animal Reproduction
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• v.10 no.1
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• pp.91-99
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• 1986

This experiment was undertaken to examine the effects of HIS treatment on the motility and acrosome reaction of frozen bovine spermatozoa and to test their abilities to interact with zona-free hamster eggs in vitro. Also, in vitro results were compared with those of bull's fertility in AI. The frozen semen from four Holstein bulls were exposed to HIS-DM for 5 minutes after thawing and then preincubated for 60 minutes in DM prior to insemination. The hamster eggs were mounted, fixed and stained 6 hours after exposure to boving spermatozoa and examined under a phase-contrast microscope. 1. The sperm motility expressed as a mobility index dro, pp.d significantly from 60-75 to 12-24 after exposure to HIS-DM, but increased in 32 to 41 at insemination. Bull C showed a low motility index than those of the orher bulls. The percentage of acrosome reaction by staining procedure were increased by HIS-DM treatment but did not change during 7 hours incubation period in DM. 2. The overall percentage of hamster eggs interacting with bull spermatozoa was 56.3%, 58.3%, 66.6% and 70.0%, respectively. Although there was no significant difference among bulls in the penetration rate of spermatozoa into hamster eggs, high proportions of eggs interacted with spermatozoa from Bull C and D than those from Bull A and B. 3. The conception rates (60-90 day RP) resulting from AI were 62.5%, 67.5% and 70.9% for Bull A, B and C, respectively. These results were in good agreement with the invitro results that the proportions of bull sperm-egg interction were greater for Bull C than for Bull A and B.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.127-134
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• 1996

The objective of this experiment was to test the ability of the fertilization of EGF treated pig oocytes for in vitro maturation. The addition of EGF (10 ng/ml), FSH (10 ${\mu}\textrm{g}$/ml), or FBS (10%) on maturation medium of pig immature oocytes divided into four groups as follows; group 1: untreatment, group 2: EGF alone, group 3: combination of FSH and FBS, or group 4: combination of EGF, FSH, and FBS. The interactive effects of nuclear maturation rates (M II%) of EGF alone, FSH plus FBS, and EGF plus FSH added FBS treatments were significantly higher than those of non-treatments (P<0.001). The fertilization rate of EGF alone (group 2) was lower than that of 3, 4 groups, but was significantly higher than group 1 (p< 0.005). Furthermore, combination of EGF, FSH,and FBS (group 4) was higher than others (group 1. 2, 3) on male pronuclei formation as well as penetration of sperm (P<0.05). These results suggested that EGF alone decreased the ability of cytoplasmic maturation compared to nuclear maturation in pig oocytes, but a high level of cytoplasmic maturation of in vitro-matured pig oocytes can be achieved when supplemented with FSH and FBS.