• Reproductive and Developmental Biology
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• v.35 no.2
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• pp.185-190
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• 2011

To evaluate the effect of spermatozoa culture on glycosidase activity of frozen-thawed spermatozoa in human, the spermatozoa were treated experimentally and assayed for activities of ${\alpha}$-L-fucosidase, ${\alpha}$-D-mannosidase, ${\beta}$-D-galactosidase and N-acetyl-${\beta}$-D-glucosaminidase (${\beta}$-GlcNAc'ase). The ${\beta}$-GlcNAc'ase activity was at least two-folds higher than other glycosidases regardless of spermatozoa incubation (p<0.05). The spermatozoa motility was decreased with incubation periods, but no effects by different glycosidases on the changes of spermatozoa motility during the various periods of incubation. In all glycosidases, the spermatozoa-zona binding rates in spermatozoa without incubation were higher than in spermatozoa incubated for 2 h (p<0.05). ${\beta}$-GlcNAc'ase is present mainly in the plasma membrane of spermatozoa frozen-thawed in human. It was also shown that the glycosidase activity was increased in all glycosidases in spite of lower sperm-zona binding by spermatozoa incubation.

• Toxicological Research
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• v.16 no.1
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• pp.47-52
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• 2000

To evaluate the renal toxicity of the antitumor agent, 1-(N-methyl) piperazinyl-3-phenyl-isoquinoline(CWJ-$\alpha$-5), rats were terated with CWJ-$\alpha$-5 (acute : 100mg/kg, i.p., single and subacute : 10mg/kr, i.p., daily for 7 days). The changes in the body weights, water consumption, kidney weights and urine volume after and during the treatment were observed. The concentrations of urinary creatinine, the activities of N-acetyl-$\beta$-D-glucosaminidase (NAG), alanine aminopeptidase (AAP), $\gamma$-glutamyl transpeptidase ($\gamma$-GT) and lactate dehydrogenase (LDH) in 24 hr urine were also determined. The body weight and water consumption were decreased after the acute and subacute administration. However, the excretion of urine was not changed except the 1 day after the acute treatment. The excretion of creatinine was significantly decreased from 1 day after acute administration and continuously decreased. Also the excretion of creatinine was decreased during subacute administration. However, the protein excretion did not changed in both treatment. Those indicate that CWJ-$\alpha$-5 might decrease the metabolic rate of muscle. The urinary activities of NAG, AAP, $\gamma$-GT, and LDH were significantly affected by the drug treatment. The urinary activities of NAG, AAP and $\gamma$-GT were significantly increased 1 and 3 days after the acute administration and then returned to the control value. However, the urinary activities of LDH were increased 7 days after acute treatment. During subacute treatment, the urinary activities of $\gamma$-GT were not changed. However, the urinary activities of NAG, AAP and LDH were only significantly increased after the third administration. These results indicate that either the high acute dose or the subacute administration with low dose of the compound might induce a temporal damage in the kidney cells.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.25-25
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• 2003

본 연구는 돼지 동결-융해 정자의 배양에 의한 체외수정능력과 glycosidase activity의 관계를 검토하였으며, 또한 돼지난자의 투명대내에서 발견된 당잔기에 대한 정자의 glycosidase 특이성을 확인하기 위하여 $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase 및 N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase)의 activity를 분석하였다. 그 결과 glycosidase activity는 동결정자의 융해 후 배양하지 않았을 때보다 2시간 배양했을 때 더 높게 나타났다. $\beta$-GlcNAc'ase의 activity는 정자 배양 유무에 관계없이 다른 glycosidase 처리시보다 최소한 2배 이상 높게 나타났다. 또한 첨체반응이 유기된 정자의 비율은 glycosidase ($\alpha$-D-mannosidase; P<0.05)에 의해 영향을 받았으며 정자를 배양하지 않은 경우보다는 배양된 정자에서 높게 나타났다. 그러나 배양시간에 따른 정자의 생존성에 대해 glycosidase의 종류에 따른 유의차는 인정되지 않았다. 한편 투명대내 정자의 접착과 침입에 대한 또 다른 실험에서, 서로 다른 glycosidase가 첨가된 배양액내에서 수정된 정자가 배양시간이 길어짐에 따라 정자의 침입율은 낮아졌다($\beta$-GlcNAc'ase; P<0.05). 투명대내의 정자접착 정도는 glycosidase의 첨가시에 무첨가시보다 접착정도가 더 높았으며, 가장 높은 접착율은 $\beta$-GlcNAc'ase첨가시 나타났다. 또한 모든 glycosidase 처리시 2시간 배양한 정자보다는 배양하지 않은 정자에서 투명대에 대한 접착정도가 높게 나타났으며, $\alpha$-D-mannosidase의 처리시 유의적인 차이를 보였다(P<0.05). 본 연구의 결과, $\beta$-GlcNAc'ase가 주로 돼지정자의 원형질막내에 존재하는 것으로 추측되며, 배양된 정자에 의한 투명대 접착정도와 침입율이 낮았음에도 불구하고 glycosidase activity가 증가하는 것으로 나타났다

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.153-161
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• 2003

This study has evaluated effect of the spermatozoa incubation on the glycosidase activity and fertilizing ability in vitro in the pig. To identify sperm glycosidases specific for sugar residues found in the zona pellucida of pig oocytes, the spermatozoa were treated experimentally and assayed for activities of $\alpha$-L-fucosidase, $\alpha$-D-mannosidase, $\beta$-D-galactosidase and N-acetyl-$\beta$-D-glucosaminidase ($\beta$-GlcNAc'ase). The glycosidases activity were higher in spermatozoa incubated for 2h than without incubation. The $\beta$-GlcNAc'ase activity was at least two-fold higher than other glycosidase regardless of spermatozoa incubation. In the same glycosidases, the activity had a tendency to increase as time of spermatozoa incubation was prolonged, but there were no differences in spermatozoa incubated during the various periods (4~24h). The percentages of spermatozoa that reached acrosome reaction were affected by glycosidases in the medium (P<0.05, for mannosidase), and were higher in spermatozoa with that than without incubation. On the other hand, the spermatozoa motility were decreased with incubation periods, but no effects by different glycosidases on the change of sperm motility during the various periods of incubation. In other experiment, the binding and penetration of pig spermatozoa were tested with oocytes matured in vitro in the presence of various glycosidase. The penetration rates were decreased with incubation of spermatozoa when oocytes were inseminated in medium with different glycosidases. These rates were higher in spermatozoa non-incubated than with incubation for 2h (P<0.05 for GlcNAc'ase; P<0.01 for control group). The sperm-zona binding rate in control group were higherthan in medium with glycosidases. In addition, the highest binding rate were obtained in medium with GlcNAc'ase. In all glycosidases, the sperm-zona binding rate in spermatozoa without incubation were higher than incubation for 2h. The significant differences were obtained in spermatozoa treated with $\alpha$-D-mannosidase (P<0.05). These results suggest that $\beta$-GlcNAc'ase is present mainly in the plasma membrane of pig spermatozoa. It was also shown that the glycosidase activity were increased in all glycosidases in spite of low sperm-zona binding rate and penetration rates by spermatozoa incubation.

• Clinical and Experimental Pediatrics
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• v.46 no.10
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• pp.977-982
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• 2003

Purpose : This study aimed to examine the excretion of various urinary proteins in children with a history of urinary tract infection(UTI), with or without vesicoureteral reflux(VUR) or reflux nephropathy, and to identify means of predicting the severity of VUR or the presence of reflux nephropathy as indicated by these markers, and to know how these markers are changed after resolution of VUR. Methods : We studied 30 children with previous UTI, without VUR and renal scarring(group I), 12 children with VUR, without evidence of renal scarring(group II), and 34 children with VUR and renal scarring(group III). 24-hour or 12-hour urine ${\beta}_2$ microglobulin(${\beta}_2$ MG), microalbumin and N-acetyl-${\beta}$-D-glucosaminidase(NAG) were measured in each child. Urinary protein excretions were analyzed according to the degree of VUR(mild VUR : a grade reflux I-III, severe VUR : a grade reflux IV-V). Cases of bilateral VUR were graded by the higher grade of reflux detected. A total of 46 children with primary VUR were followed. Among these patients, VUR was completely resolved in 16 children. Voiding cystourethrography(VCUG) and DMSA scan were performed every year. Values for urinary markers were estimated every year. Results : 24 or 12 hour urine microalbumin and NAG excretions were significantly increased in group III compared to group I(microalbumin : $27.7{\pm}26.0mg/gCr$ vs $15.0{\pm}10.7mg/gCr$, P<0.05, NAG : $15.2{\pm}18.7U/gCr$ vs $3.4{\pm}2.2U/gCr$, P<0.05). Urinary ${\beta}_2$ MG excretions were not significantly different between groups. Urinary NAG excretions were elevated in the group of children with severe VUR compared to mild VUR($26.8{\pm}27.1U/gCr$ vs $7.6{\pm}3.8U/gCr$, P<0.05). After resolution of VUR, urinary microalbumin and NAG excretions were decreased(P<0.05). Conclusion : Urinary microalbumin and NAG may be useful clinical indicators to predict the presence of reflux nephropathy and the resolution of VUR. Especially, urinary NAG excretions may be used as a possible method to predict the severity of VUR.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.43-43
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• 2001

This study evaluated the effect of fertilization-promoting peptide (FPP) on fertilizing ability and glycosidase activity in vitro of spermatozoa frozen-thawed in pig, Using chlortetracycline fluorescence analysis, the various glycosidase analyses and the oocyte penetration test, we have obtained evidence that FPP can promote the fertilizing ability and glycosidase activity of frozen-thawed spermatozoa in vitro. When frozen-thawed spermatozoa was washed with different concentrations of FPP, there were significantly (P<0.05) more acrosome-reacted in medium with 100 nM than 0, 50, 200 and 400 nM. The penetration rates were also highest in medium containing with 100 nM FPP (P<0.05). On the other hand, the $\beta$-N-acetylglucosaminidase activity was at least twofold higher than other glycosidase. In same glycosidase, however, there were no difference in medium with different concentrations of FPP In another experiment, spermatozoa preincubated in medium with or without FPP for 0, 1, 2, 3 and 4 h were inseminated with oocytes matured in vitro. The percentages of spermatozoa that reached acrosome reaction were affected by preincubation and were higher in medium with that than without FPP. When oocytes were inseminated with spermatozoa preincubated in medium with and without FPP during the different periods, however, penetration rates were decreased with preincubation periods of spermatozoa. On the other hand, when the sperm-oocyte were cultured for 4, 8, 12, 16, 20 and 24 h, the penetration rates were higher in spermatozoa preincubated with that than without FPP and had a tendency to increase as time of culture periods. However, The activities of $\alpha$-fucosidase, $\alpha$ -mannosidase, $\beta$-galactosidase and N-acetyl- $\beta$-D-glucosaminidase were higher in medium with that than without FPP regardless of periods of sperm preincubation and sperm-oocyte culture. These results suggest that FPP may play a positive role in promoting of sperm function and glycosidase activity in vitro in pig.

• Toxicological Research
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• v.32 no.1
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• pp.57-64
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• 2016

Recently several studies reported that the renal toxicity of lead (Pb) and cadmium (Cd) may exist in even a low level exposure. In terms of the deterioration of tubular function, it affects the loss of divalent metals and leads to other complications, so renal tubular effect of heavy metals should be well managed. Considering the exposure to heavy metals in reality, it is hard to find the case that human is exposed to only one heavy metal. We designed a cross-sectional study using Korean Research Project on the Integrated Exposure Assessment (KRIEFS) data to investigate the renal effects of multiple metal exposure in general population. We used blood Pb and urinary Cd as exposure measures, and urinary N-acetyl-${\beta}$-D-glucosaminidase (NAG) and ${\beta}_2$-microglobulin (${\beta}_2$-MG) as renal tubular impairment outcome. We conducted linear regression to identify the association between each heavy metal and urinary NAG and ${\beta}_2$-MG. And then, we conducted linear regression including the interaction term. Of 1953 adults in KRIEFS (2010~2011), the geometric mean of blood Pb and urinary Cd concentration was $2.21{\mu}g/dL$ (geometric $SD=1.49{\mu}g/dL$) and $1.08{\mu}g/g\;cr$ (geometric $SD=1.98{\mu}g/g\;cr$), respectively. In urinary Cd, the strength of the association was also high after adjusting (urinary NAG: ${\beta}=0.44$, p < 0.001; urinary ${\beta}_2$-MG: ${\beta}=0.13$, p = 0.002). Finally, we identified the positive interactions for the two renal biomarkers. The interaction effect of the two heavy metals of ${\beta}_2$-MG was greater than that of NAG. It is very important in public health perspective if the low level exposure to multiple heavy metals has an interaction effect on kidney. More epidemiological studies for the interaction and toxicological studies on the mechanism are needed.

• Food Science of Animal Resources
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• v.34 no.6
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• pp.742-748
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• 2014

This study was conducted to evaluate the activities of antioxidant enzyme (glutathione peroxidase (GSH-Px)) and lysosomal enzymes (alpha-glucopyranosidase (AGP) and beta-N-acetyl-glucosaminidase (BNAG)) of the longissimus dorsi (LD) muscle from Hanwoo (Korean cattle) in three freezing conditions. Following freezing at -20, -60, and $-196^{\circ}C$ (liquid nitrogen), LD samples (48 h post-slaughter) were treated as follows: 1) freezing for 14 d, 2) 1 to 4 freeze-thaw cycles (2 d of freezing in each cycle), and 3) refrigeration ($4^{\circ}C$) for 7 d after 7 d of freezing. The control was the fresh (non-frozen) LD. Freezing treatment at all temperatures significantly (p<0.05) increased the activities of GSH-Px, AGP, and BNAG. The $-196^{\circ}C$ freezing had similar effects to the $-20^{\circ}C$ and $-60^{\circ}C$ freezing. Higher (p<0.05) enzymes activities were sustained in frozen LD even after 4 freeze-thaw cycles and even for 7 d of refrigeration after freezing. These findings suggest that freezing has remarkable effects on the activities of antioxidant enzyme and lysosomal enzymes of Hanwoo beef in any condition.

• Journal of Veterinary Clinics
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• v.7 no.2
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• pp.471-487
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• 1990

Present experiment was performed in order to establish the optimum conditions for quantitation of ${\gamma}$-GTP and AGS activities in rat urine and investigate the applicability of the these enzymes in experimental assessment of nephrotoxicity in rats. The results obtained were as follows. 1. The optimal pH of Tris-BCI buffer containing glycylglycine for determination of urinary ${\gamma}$-GTP activity was 7.6(37$^{\circ}C$). 2. The Michaelis constant of urinary ${\gamma}$-GTP ranged from 1.1 to 1.2 mmol/$\ell$. 3. The optimal pH of citrate buffer for determination of urinary AGS activity was 3.6(37$^{\circ}C$). 4. The Michaelis constant of urinary AGS ranged from 0.8 to 0.9mmo1/$\ell$. 5. Coefficient of variance for within-run imprecision of urinary ${\gamma}$-GTP ranged from 3.8 to 6.4％ and that of urinary AGS ranged from 2.5 to 4.1％. 6. There was no significant difference between gel-filtered samples and crude samples in the mean activity of urinary ${\gamma}$-GTP and the intra-individual differences by gel-filtration were either increased or decreased. Mean values of ${\gamma}$ -GTP activities in gel-filtered samples and crude samples were 1570 and 1590 U/$\ell$, repectively. 7. The mean activity of urinary AGS increased significantly after gel-filtration and all the individual urines revealed higher activities after gel-filtration. 8. ${\gamma}$-GTP and AGS activities were linear to 135 and 7U/$\ell$, respectively. 9. Urinary ${\gamma}$-GTP and AGS excretion before administration of potassium dichromate were 22.1 ${\pm}$ 11.2 and 0.5${\pm}$0.2 U/24hrsㆍkg body weight respectively and increased significantly to 102.3${\pm}$44.5 and 5.8${\pm}$3.30/24hrsㆍkg body weight respectively within 24 hours after administration. 10. BUN increased continuously from 24 hours following exposure to potassium dichromate in all 10 rats. From these findings it is concluded that the urinary ${\gamma}$-GTP and AGS excretions are early and sensitive indicators for nephrotoxicity assessment in rat.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.213-213
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• 1996

The effects of the anti-tumor agent, SDZ-y2434, on rat kidney were investigated to predict the toxicities of its derivatives and to develope less toxic derivatives. After adjusted in metabolic cages for 5 days, rats were treated SDZ-62434(acute : 25mg/kg, i.p, once and 50mg/kg, i.p., once; subacute ; 10mg/kg, i.p., daily for 7 days). Kidney weights and urine volume during the treatment were observed. Creatinine concentration, protein concentration and the activities of N-acetyl-${\beta}$-D-glucosaminidase (NAG), alanine aminopeptidase (AAP), ${\gamma}$-glutamyl transpeptidase (GGT) and lactate dehydrogenase(LDH) in 24 hr urine were also determined. The kidney weights after the acute and subacute administration didn't show any difference. Urine volume increased 5 days after the acute administration (50mg/kg) and 3 days after the subacute administration. The excretion of creatinine was increased 5 days after the acute (50mg/kg) and subacute administration. However, the protein excretion didn't show any change. NAG acivity declined 7 days after the subacute administration. AAP and GGT activites increased 3 days after the acute administration (50mg/kg) but, returned to the control value. LDH activity showed continuousely high value after the subacute administration. These results indicates that the acute administration of SDZ-62434 might damage on glomerulus and that the subacute administration might be cytotoxic to kidney cells.