• Korean Journal of Poultry Science
• /
• v.40 no.3
• /
• pp.171-178
• /
• 2013

The purpose of this study was to evaluate the sperm viability, normal acrosome and mitochondrial activity in the frozen-thawed fowl semen by different cryoprotectants. The experiment was carried out on 10 sexually adult roosters of Ogye. The semen was collected twice a week and pooled semen was diluted 1:1 EK extender containing no cryoprotectant at $5^{\circ}C$. After equilibration for 30 minutes, diluted chicken semen was diluted 1:1 extender containing either 7% dimethylacetamide (DMA), 7% dimethylformamide (DMF) or 7.5% methylacetamide (MA) at final concentration and was put in 0.5 mL plastic straws and frozen for 30 minutes by exposure to liquid nitrogen vapor 4 cm above the surface of liquid nitrogen, followed by plunging into liquid nitrogen. Frozen semen was thawed in water bath at $5^{\circ}C$ for 2 minutes. For cytometric analysis, the frozen-thawed semen was diluted with EK extender to a final concentration of 90 million spermatozoa per mL. Sperm membrane integrity was evaluated as SYBR-14 and propidium iodide (PI). Acrosome integrity was assessed with fluorescein isothiocyanate-labeled PSA and PI. The percentage of mitochondrial function was estimated by using Rhodamine123 (R123) and PI. In conclusion, freezing rooster semen by using 7% DMF as cryoprotectant was significantly highest in rates of survival and mitochondrial function while its rate of damage of acrosome was significantly lowest. As a result, DMF is the cryoprotectant that has the lowest influences on sperm membranes and acrosome integrity. Therefore it could be used for freezing method of animal genetic conservation method for poultry diversity.

• Journal of Korean Ophthalmic Optics Society
• /
• v.20 no.2
• /
• pp.141-150
• /
• 2015

Purpose: In the present study, a difference in tear volume between the cornea and the rigid gas permeable (RGP) lens relative to corneal shape and corneal astigmatism was investigated by the alignment fitting status of spherical and aspherical RGP lenses. Methods: Spherical and aspherical RGP lenses were fitted with alignment in 77 subjects (135 eyes) who were in their 20~30s. Tear volume stained with fluorescein was qualitatively analyzed by dividing cornea into center, mid-peripheral and peripheral parts. Results: For the spherical RGP lens fitting, tear volume differences were found in each part in all corneal types. For the aspherical RGP lens fitting, tear volume differences were in each corneal part in symmetric bow tie- and asymmetric bow tie-type corneas. However, the tear was equally distributed from the center to the peripheral part in round- and oval-type corneas. In the group with corneal astigmatism lower than 1.25 D, tear volume between center and peripheral parts, and mid-peripheral and peripheral parts, was different when a spherical RGP lens was fitted. However, tear volume in each part was not different in the group with corneal astigmatism over 1.50 D. Moreover, the tear volumes of the central and mid-peripheral parts were proportionally increased with increasing corneal astigmatism in both spherical and aspherical RGP lenses. Furthermore, aspherical RGP lenses showed greater increments than spherical RGP lenses. Conclusions: The results revealed that the difference in tear volume between aspherical RGP lens and cornea was less than spherical RGP lens, and the difference in tear volume varied according to corneal shape and astigmatism. In addition, the method of measuring relative tear volume between RGP lens and cornea that was established in the present study can be used to evaluate tear volume between contact lens and cornea.

• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.3
• /
• pp.311-318
• /
• 1997

Bypassing acrosome reaction and fusion process in intracytoplasmic sperm injection(ICSI), most of injected spermatozoa still contain intact acrosome contents and plasma membrane. It Is not known yet what acrosome contents and plasma membrane of spermatozoa have effect on the development of embryo. For further understanding of fertilization process after ICSI, we studied the time of pronucleus formation, disappearance and first cleavage in human zygote, and pregnancy rate in relation to acrosome reaction rate of spermatozoa after ICSI. Seventy cycles undergoing ICSI program were randomly selected. Sperm suspension from 38 cycles were treated 50% human follicular fluid(hFF) for 3 hours in order to induce acrosome reaction, others were not treated as control. Acrosome reaction in hFF treated and non-treated group was assessed by fluorescein isothiocyanate(FITC)-conjugated Arachis hypogea(PNA) and Pisum sativum agglutinin(PSA). Oocytes were classified into 'good' and 'poor' according to their morphology. After ICSI, fertilization of oocytes were assessed by detection of two pronuclei at 16 hours. The pronuclei disappearance and first cleavage of zygotes were observed at 24 hours, and then embryos were transferred to uterus after culture for 72 hours. The rate of acrosome reaction of spermatozoa in hFF treated group was significantly higher than that in control(p<0.01). Fertilization rates of good oocytes were not different both control and hFF treated group(81.3%(174/206) vs. 72.1%(102/130)). But, in poor oocytes, the fertilization rates in hFF treated group(72.1%(149/183)) were increased compared than those of control group (63.6%(98/140), p<0.01). In either good or poor oocytes, the rates of pronuclei disappearance in hFF treated-spermatozoa injected oocytes were higher than control (59.1%(103/174), 56.4%(84/149) vs. 32.4%(33/102), 37.8%(37/98), p<0.01). Also, the rates of thirst cleavage were increased in hFF treated group (31%(54/174), 24.1%(36/149)) compared than those of control group (10.8%(11/102), 13.2%(13/98), p<0.01). The pregnancy rates of hFF treated group (42.1%(16/38)) were slightly higher than control group (28.1%(9/32), p>0.05). But, the pregnancy rate of group which possessed more than one cleavaged zygote at 24 hours was higher than group which did not (45.2%(19/42) vs. 21.4%(6/28), p<0.05). From these results, the development of zygotes were faster in higher acrosome reacted sperm group than lower acrosome reacted sperm group after ICSI. Our results may be explained that acrosomal membrane and plasma membrane are easily detached from spermatozoa in acrosome reacted spermatozoa compared with acrosome intact sperm in the cytoplasm of oocyte during pronuclear formation. We conclude that the injection of acrosome reacted spermatozoa will increase the pregnancy rate as they can induce fast embryonic development in ICSI.

• KSBB Journal
• /
• v.18 no.1
• /
• pp.55-61
• /
• 2003

To isolate and purify the antimicrobial and antitumor agents in Xanthium strumarium L. hydrothermal extract. The crude extract was extracted in ether or ethylacetate under neutral, acidic, and alkali conditions. The antimicrobial activity of each extract was tested against 16 strains of bacteria, 2 strains of yeast, and 2 strains of fungus. The ether neutral extract (XE-N) exhibited the strongest growth inhibition upon the 8 strains of gram-positive bacteria, 6 strains of gram-negative bacteria and Cryptococcus neoformans. Fluorescein diacetate (FDA) testing of XE-N and XEA-N showed growth inhibition of the 3 strains of E. coli, S. aureus and C. albicans even at 30 ng/mL, with the exception of p. aeruginosa. XE-N-S1 and XE-N-S3 from neutral ether extract (XE-N), XE-N-S3 from the acidic ether extract (XE-A), and XEA-N-S1 from ethylacetate (XEA-N) were purified as antimicrobial and antitumor agents. However all purified compounds decomposed with the exception of XE-N-S1. The results upon the antitumor activities of the crude extract and of its purified compounds, showed that XE-N-S1 had the best antitumor activity against HeLa cells. In terms of antitumor activity against HepG2 cells, XE-N-S1 and XE-N-S3 were superior, and against HT29 cells XE-N and XE-N-Sl were good, against Saos2, NCI H522, NCI H1703, Clone M3 cells XE-N-51 was very good, and against LN CAP cells XE-N-S3 was the best. Comparing of cellular toxicities various extracts and purified compounds with the existing antitumor agents, XE-A, XEA-A and XEA-B had the lowest toxicity, and XE-B had a lower toxicity than etoposide. XE-N-S1 and XE-N-S3 showed higher toxicities than etoposide, and the toxicity of XE-A-S3 was higher than that of etoposide, and lower than that of csplatin.

• Restorative Dentistry and Endodontics
• /
• v.24 no.2
• /
• pp.310-321
• /
• 1999

In this in vitro study, confocal laser scanning microscopic morphology of dentin-resin interface and its relationship to shear bond strength were investigated after the exposed dentin surfaces were treated with 3 different kinds of dentin adhesive systems[three-step; Scotchbond Multi-Purpose Plus(SMPP), self-priming bonding resin; Single Bond(SB), self-etching primer; Clearfil Liner Bond 2(LB2)]. 52 extracted human molar teeth without caries and/or restorations. The experimental teeth were randomly divided into three groups of seventeen teeth each. In five teeth of each group, class V cavities(depth: 1.5mm) with 900 cavosurface angles were prepared at the cementoenamel junction on buccal and lingual surfaces. Bonding resins of each dentin adhesive system were mixed with rhodamine B. Primer of SMPP was mixed with fluorescein. In group 1. the exposed dentin was conditioned with etchant, applied with above primer and bonding resin of SMPP. In group 2, with etchant and self-priming bonding agent of SB. In group 3, with self-etching primer and bonding agent of LB2. After treatment with dentin adhesive systems, composite resin were applied and photocured. The experimental teeth were cut longitudinally through the center line of restoration and grounded so that about $90{\mu}m$-thick wafers of buccolingually orientated dentin were obtained. And, $70{\sim}80{\mu}m$-thick wafers sectioned horizontally, thus presenting a dentinal tubules at 900 to the cut surface of a remaining tooth, were obtained. Primer of SMPP mixed with rhodamine B was applied to these wafers. Confocal laser scanning microscopic investigations of these wafers were done within of 24 hours after treatment. To measure shear bond strength, the remaining twelve teeth of each group were grounded horizontally below the dentinoenamel junction, so that no enamel remained. After applying dentin adhesive systems on the dentin surface, composite was applied in the shape of cylinder. The cylinder was 5mm in diameter, and 2mm in thickness. Shear bond strength was measured using Instron with a cross-head speed of 0.5mm/min. It was concluded as follows ; 1. Hybrid layer of SMPP(mean: $4.56{\mu}m$) was thicker than that of any other groups. This value was not statistically significant thicker than that of SB(mean: $3.41{\mu}m$, p>0.05), and significant thicker than that of LB2(mean: $1.56{\mu}m$, p<0.05). There was a statistical difference between SB and LB2(p<0.05). 2. Although there were variations in the length of resin tag even in a sample, and in a group, most samples in SMPP and SB showed resin tags extending above $20{\mu}m$. But samples in LB2 showed resin tags of $10{\mu}m$ at best. 3. Besides primer's infiltration into demineralized peritubular dentin and dentinal tubules, fluorophore of primer was detected in the lateral branches of dentinal tubules. 4. All groups demonstrated statistically significant differences from one another(p<0.05), with shear bond strengths given in descending order as follows: SMPP(18.3MPa), SB(16.0MPa) and LB2(12.4MPa). 5. LB2 having thinnest hybrid layer($1.56{\mu}m$) showed the lowest shear bond strength(12.4MPa).