• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.105-109
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• 2003

Objective s: To assess the fertilizing capacity using sperm penetration assay (SPA) to predict the outcome of the in vitro fertilization-embryo transfer (IVF-ET) outcome. Materials and Methods: Semen samples were provided by 129 patients undergoing IVF. We attempted to correlate the extent of sperm penetration under enhanced SPA protocol with the results of fertilization, cleavage, preimplantation embryo development, and pregnancy. Results: Univariate analysis demonstrated a statistically significant correlation between fertilizing capacity and motility, kinetics, fertilization, cleavage and embryo development, and pregnancy rate. By logistic regression analysis, fertilizing capacity was found to be the only variable that was statistically significant with respect to pregnancy rate. Fertilizing capacity, cleavage rate and pregnant rate were significantly higher in pregnant group. However, the fertilization rates was comparable with both group. Conclusions: Lower fertilizing capacity could denote a poorer prognosis for establishing a pregnancy, even after satisfactory fertilization rate is achieved.

• Journal of Pharmaceutical Investigation
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• v.33 no.3
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• pp.157-162
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• 2003

The feasibility of [P(AA-co-PEGMM)] film as a buccal mucoadhesive patch was previously reported by estimating mucoadhesiveness and release characteristics. To find a rational penetration enhancer of [P(AA-co-PEGMM)] film containing butorphanol tartrate (Bt), penetration of Bt from [P(AA-co-PEGMM)] film which contained various additives was estimated by measuring its flux, Papp and lag tme in in vitro buccal membrane of porcine. EDTA showed almost no increase of Bt permeability, wherease SGC, STDHF and SLS increased the permeability of Bt with the order of SGC > STDHF > SLS. The rational additive concentration of SGC was 4% and its Papp and lag time were $1.93{\times}10^{-4}{\pm}4.21{\times}10^{-6},\;126.60{\pm}21.88min\;(control\;:\;Papp\;0.45{\times}10^{-4};\;lag\;time\;211.01{\pm}16.77\;min)$, respectively.

• YAKHAK HOEJI
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• v.58 no.3
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• pp.171-180
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• 2014

To enhance the in vitro permeation of lovastatin through excised hairless mouse and human cadaver skins, solubility was determined in various hydrophilic and lipophilic vehicles, and the effects of vehicles and penetration enhancers on the skin permeation from solution formulations were investigated. Solubility of lovastatin was highest in N-methyl-2-pyrrolidone (NMP) ($278.2{\pm}10.1$ mg/ml) and dimethyl sulfoxide (DMSO) ($162.2{\pm}9.7$ mg/ml). Among different pure vehicles used, NMP, DMSO, propylene glycol and isopropyl myristate provided some drug permeation ($6.9{\pm}1.1$, $5.9{\pm}1.6$, $3.0{\pm}0.5$ and $2.2{\pm}0.3{\mu}g/cm^2$ at 24 hr, respectively) through hairless mouse skin. The addition of oleic acid, linoleic acid and oleyl alcohol to DMSO showed the maximum permeation at around 5 v/v%, however, capric acid and caprylic acid had no enhancing effect. The increase of enhancer concentrations showed bell-shaped permeation rate, suggesting the presence of optimal concentration in lovastatin penetration. Increasing donor concentration from 10 mg/ml to 80 mg/ml in DMSO and a cosolvent of DMSO, NMP and DGME (3 : 3 : 4 v/v) did not show significant dose dependent permeation in both hairless mouse and human cadaver skins. The maximum lovastatin flux through human cadaver skin was found to be $0.87{\pm}0.46{\mu}g/cm^2$/hr with 5 v/v% linoleic acid and donor dose of 4 mg/0.64 $cm^2$ in the cosolvent. These results suggest that transdermal delivery of lovastatin would be feasible by establishing the optimal concentrations of donor dose and unsaturated fatty acids in appropriate vehicles.

• YAKHAK HOEJI
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• v.44 no.6
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• pp.595-600
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• 2000

Transdermal delivery of ketorolac tromethamine, a potent non-narcotic analgesic, through human cadaver skin was investigated in vitro. A mixture of ethanol/water (40/60) containing 0, 1, 3, 5, and 8 (w/v)% L-menthol were used as a vehicle and penetration enhancer respectively. The permeation of ketorolac through human cadaver skin from saturated drug solution was evaluated at $37^{\circ}C$ with modified Franz diffusion cell. The in vitro skin flux and lag time were $1.23\;{\pm}\;0.11\;{\mu}g/cm^2{\cdot}hr$ and $5.56\;{\pm}\;0.34\;hr$, respectively. The cumulative amount of penetrated ketorolac containing L-menthol in ethanol/water (40/60) binary system was increased by the following order; 3%, 5%, 8%, 1%, 0%, and the lag time was decresed by the following order; 3%, 5%, 8%, 0%, 1%. The results suggested that a potential use of 3% L-methol is an effective penetration enhancer of ketorolac tromethamine through the human cadaver skin.

• Reproductive and Developmental Biology
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• v.31 no.2
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• pp.115-120
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• 2007

This study was conducted to examine the effect of several saccharides on the induction of capacitation and acrosome reaction (AR) and to examine the effects of mono and polysaccharides on the penetration activity of boar spermatozoa. Spermatozoa were inseminated in medium with fucose, galactose and mannose as monosaccharide, and fucoicIan. galactan and marman as polysaccharide. The penetration rates were significantly (p<0.05) lower in medium with galactose (40.6%), mannose (38.1%), fucose (41.6%) and fucoidan (36.6%) compared with control (56.7%). The rates of AR were increased (40.7 to 59.8%) by the preincubation periods prolonged from 0 to 4 hr (p<0.05). Similar tendencies were observed in AR when spermatozoa were treated with monosaccharides, but not significantly differ among the groups treated with different time of preincubation with some exception of galactose. When spermatozoa were treated with polysaccharides, the rates of AR were significantly (p<0.05) increased by preincubation time prolonged from 0 to 4 hr with an exception of fucoidan. In conclusion, the present study suggests that penetration rate of spermatozoa is higher in presence of polysaccharides than monosaccharides. Also, it may resume that the comparing to control, the all saccharides (L-fucose, D-galactose, D-mannose, fucoidan. galactan and mannan)-treated groups slightly increase the AR pattern as preincubation time prolonged.

• Korean Journal of Animal Reproduction
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• v.20 no.1
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• pp.1-8
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• 1996

The objective of this study was to determine effects of oviductal fluid on the sperm penetration and subsequent in vitro development of porcine oocytes. The addition of oviductal fluid to the fertilization medium decreased sperm pen etration and the mean number of spermatozoa in penetrated eggs. The number of spermatozoa firmly bound to zona pellucida was also decreased in the presence of oviductal fluid. Chlortetracy cline (CTC) fluorescence patterns were used to determine incidence of capacitation and acrosome reaction. The proportion of capacitated a and acrosome free spermatozoa increased when spermatozoa were exposed for 1.5 and 3 h to oviductal fluid. These results suggest that the factor(s) in secretion from the oviduct reduces polyspermic fertilization and the number of spermatozoa that will penetrate porcine oocytes. The reduction of polyspermic penetration by oviductal secretions may be due to a reduced number of spermatozoa in the fertilization me-dium into an intact acrosome.

• Korean Journal of Animal Reproduction
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• v.27 no.4
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• pp.275-280
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• 2003

This study was carried out to investigate the effect of cysteamine addition during in vitro maturation, fertilization and culture of porcine oocytes. Oocytes were matured for the first 22 h in mTCM -199 media supplemented with or without 150 $\mu$M cysteamine. They then were matured for an additional 22 h in mTCM-199 media without hormones supplemented with or without 150 $\mu$M cysteamine. When cumulus-oocyte complexes (COCs) were matured in the mTCM-199 media supplemented with cysteamine, the rates of GVBD and maturation (metaphase II) were enhanced as compared to the media without the addition of cysteamine. Also, when COCs were matured in the mTCM-199 media supplemented with cysteamine, the rates of sperm penetration, male pronucleus formation, cleavage and blastocyst formation after in vitro fertilization were enhanced as compared to the media without the addition of cysteamine. In conclusion, it was suggested that oocytes matured for the first 22 h in mTCM-199 media supplemented with 150 $\mu$M cysteamine increased the rates of metaphase II, sperm penetration, male pronucleus and blastocyst formation were higher as compared to the media without addition of cysteamine.

• Biomolecules & Therapeutics
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• v.14 no.2
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• pp.90-95
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• 2006

CKD-602 exerts its antitumor effect via inhibition of topoisomerase I in cancer cells. Multicellular spheroid (MCS) and Multicellular layers (MCLs) are known as in vitro 3-dimensional models which closely represent tumor conditions in vivo. In order to investigate the potential of CKD-602 against human colorectal tumors, we evaluated the anti-proliferative activity and penetration ability of CKD-602 in MCS and MCL cultures of DLD-l human colorectal cancer cells, respectively. The maximum effects($E_{max}$) induced by CKD-602 were significantly lower in MCS compared to monolayers (48% vs 92%). With prolonged drug exposure, the $IC_{50's}$ of CKD-602 decreased to $23.5{\pm}1.0nM$ in monolayers after 24 h exposure and $42.3{\pm}1.7nM$ in MCS after 6 days, respectively. However, no further increase in effect was observed for exposure time longer than growth doubling time (Td) in both cultures. Activity of CKD-602 was significantly reduced after penetration through MCL and also with cell-free insert membrane. In conclusion, CKD-602 showed significantly decreased anti-proliferative activity in 3D cultures (MCS) of human colorectal cancer cells. Tumor penetration of CKD-602 could not be determined due to loss of activity after penetration through cell free insert membrane, which warrants further evaluation using a modified model.

• Reproductive and Developmental Biology
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• v.33 no.3
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• pp.125-131
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• 2009

The aim of this study is to investigate the effect of porcine epididymal fluid (pEF) on in vitro-maturation and subsequent fertilization of porcine follicular oocytes. Porcine cumulus-oocytes complexes retrieved from antral follicles were cultured in tissue culture medium (TCM)-l99 supplemented with pEF of different concentrations. At 48 h after culture, development of oocytes to germinal vesicle (GV) breakdown, metaphase I, anaphase-telophase I, and metaphase II were examined Significant (p<0.05) increase in the proportion of oocytes developed to MII stage was observed in oocytes cultured in pEF-containing TCM-l99 than in oocytes cultured in pEF-free TCM-199 (46.2% vs 16.7%), which was a dose-dependent manner. Subsequently, the proportion of monospermic fertilization were significantly (p<0.05) increased in oocytes cultured in the TCM supplemented with pEF than those cultured in pEF-free TCM-199 (51.0% vs 24.1%). In the second series of experiment, the percentage of MII oocytes was significantly (p<0.05) increased after exposure of oocytes to pEF during the first 22 h period of culture than after exposure of oocytes to pEF during the next 24 h of culture, while no significant difference in the percentage of monospermy was observed. The results of this study demonstrate that pEF contains at least enhancing component(s) for nuclear maturation.

• Journal of Pharmaceutical Investigation
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• v.26 no.1
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• pp.43-53
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• 1996

The objective of this study was to determine whether 4-phenylazobezyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (Pz-peptide), an enhancer of hydrophilic solute permeability in the intestine, could elevate the paracellular permeability of hydrophilic drugs across cornea and conjunctiva in the rabbit. The in-vitro penetration of hydrophilic drugs (mannitol, atenolol) and lipophilic drug (propranolol) across the rabbit cornea and conjunctiva was studied either in the presence or absence of 3 mM Pz-peptide. Drug penetration was evaluated using the modified Ussing chamber. The conjunctiva was more permeable than the cornea to all drugs. Pz-peptide showed enhanced effects on the drug transport across cornea and conjunctiva in a concentration dependent manner. Effects or ion transport inhibitor on the mannitol penetration were then investigated. Mannitol penetration was not changed by serosal addition of $100\;{\mu}M$ ouabain, suggesting that $Na^+/K^+$ ion tranporter was not involved in the Pz-peptide induced elevation of paracellular drug permeability. Furthermore, effects of Pz-peptide and EDTA on the transport of atenolol and propranolol into the ocular tissues or blood circulation after its administration into both eyes were investigated. EDTA showed enhanced effect on propranolol transport into the ocular tissues, but Pz-peptide did not show significant difference. Systemic absorption of propranolol by the addition of EDTA or Pz-peptide was not changed. On the other hand, EDTA and Pz-peptide elavated the atenolol transport into the ocular tissues. The transport of atenolol into the blood circulation was also enhanced by the addition of EDTA, but no effect was observed by the addition of Pz-peptide. The above findings suggest that Pz-peptide would be used as an paracellular pathway enahncer of hydrophilic drugs into the eye, without affecting the systemic absortion of topically applied opthalmic drugs.