• 대한미생물학회지
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• 제22권3호
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• pp.233-240
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• 1987

The amniotic fluid provides a medium in which the fetus can readily move, cushions him against possible injury and helps him maintain an even temperature. Besides above mentioned functions, investigators reported that human amniotic fluid contains host-resistance factors which prevent bacteria from producing infectious disease and this activity shows difference among human racial groups or bacterial genera, species and strains. 40 amniotic fluid specimens from Korean women in their second and third trimesters of pregnancy were examined for inhibiting the growth of Escherichia coli. And various factors which might affect bacterial growth inhibiting activity such as pH, initial inoculum size, concentration of amniotic fluid, and heat resistance, were also tested using a strongly inhibitory amniotic fluid specimen. Finally plate diffusion tests were carried out using other strongly inhibitory amniotic fluid. The following results were obtained: 1. Of the 40 fluid samples examined, 18 specimens(45%) had inhibitory activity and samples from women in their second trimester of pregnanancy showed non-inhibitory activity(2 specimens). 2. The pH of the fluids varied between 7.43 and 8.33. There was no correlation between pH and inhibitory activity. 3. No. 19 amniotic fluid showed bacteriostatic activity after 24 hours incubation when an inoculum of $10^2$ organisms per milliliter was used, but non-inhibitory with an inoculum of $10^3$ and $10^4$ bacteria per milliliter. 4. The content of amniotic fluid in culture media influenced E. coli growth. At 90 percent, E. coli was inhibited growth but at 10 percent and 50 percent. 5. Inhibitory activity of No. 19 amniotic fluid was retained after heating to $50^{\circ}C$ for 30 minutes or 100^{\circ}C$ for 30 minutes. 6. Plate diffusion tests with No. 27 amniotic fluid showed that 0.7ml amniotic fluid gave clear zone of growth inhibition around the central well but 0.2ml and 0.1ml amniotic fluids were not.

• Advances in pediatric surgery
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• 제15권1호
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• pp.1-10
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• 2009

최근에 양수에 존재하는 세포들은 다양한 세포분화의 능력을 가지고 있으며 세포 치료와 조직공학의 세포원으로서의 가능성이 높다는 연구 결과가 많이 보고되고 있다. 그러나 양수 내에 어떤 종류의 세포가 정상적으로 존재하는 지에 대해서는 제한적으로 알려져 있다. 본 연구에서는 사람의 양수 내에 호흡기세포를 분리 배양하는 것을 목표로 하였다. 임신 17주에서 20주 사이의 산모 10명에서 각각 5 mL의 양수를 획득하여 small airway growth medium (SAGM) 배양액에서 계대 배양을 시행하였으며 type II alveolar cells이 존재하는 지에 대하여 면역형광염색 및 RT-PCR을 시행하였다. 배양된 세포는 광학현미경 상 상피세포와 유사한 다면체의 모양이었으며 계대 배양 시에 변화 없이 동일한 형체를 보였다. 배양된 세포는 면역형광염색 상 type II alveolar cell의 특이표지자인 surfactant protein C (SPC) 및 TTF-1 protein에 대해서는 양성이었으나 CD 31 및 vimentin에 대해서는 음성이었으며, RT-PCR 상 SPC mRNA 가 발현되었다. 이상의 결과로 사람의 양수를 특이 배양액에 배양하면 호흡기세포를 분리, 확인할 수 있음을 알았으며 이러한 결과가 향후 양수세포의 추가적인 연구에 활용될 수 있다고 생각된다.

• Archives of Pharmacal Research
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• 제20권3호
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• pp.218-224
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• 1997

Platelet-activating factor (PAF) acetylhydrolase, which removes the acetyl moiety at the sn-2 position, has been found in human amniotic fluid. We purified this enzyme by ammonium sulfate precipitation, and sequential use of DEAE-Sepharose CL-6B, hydroxyapatite, chelating-Sepharose, and Mono Q column chromatographies. This enzyme exhibited broad pH optima and was unaffected by EDTA. Partially purified enzyme had a molecular weight of approximately 34 kDa on SDS-PAGE. In addition, the enzyme activity was inhibited by either diisopropylfluorophosphate(DFP) or p-bromophenacylbromide (p-BPB), suggesting that this enzyme possesses active serine and histidine residues. The enzyme showed similar activity towards PAF and oxidatively modified phosphatidylcholine, but didn't hydrolyze phosphatidylcholine or phosphatidylethanolamine with a long chain fatty acyl group at sn-2 position.

• 한국발생생물학회지:발생과생식
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• 제16권2호
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• pp.95-100
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• 2012

Bisphenol A(BPA)는 약한 estrogen 활성도를 보이는 화학물질로서 착상 전 배아나 태아에 영향을 미쳐, 출생 후 그들의 신체발달에 변화를 초래한다고 알려져 있다. 본 연구는 인체에서 임신 중기에 양수 내 BPA 농도를 측정하였으며, 측정된 농도에 따라 임신결과에 영향을 미치는지 여부를 알아보고자 하였다. 임신 15주에서 20주 사이에 유전학적 적응증으로 양수천자 검사를 시행 받았던 120명의 임신부 양수를 연구대상으로 ELISA 방법으로 농도를 측정하였다. 양수천자의 가장 많은 적응증은 35세 이상의 고령 임신부였다. BPA는 모든 양수에서 검출이 가능하였으며, 그 농도는 최저치 0.89 ng/mL부터 최고치 37.13 ng/mL까지의 분포를 보였고, 평균치는 7.24 ng/mL이었다. 양수 내 BPA 농도가 여러 임신결과들과 관련성이 있는지를 알아보기 위해 임신부 나이 35세 이상 대 미만, 남자 태아 대 여자 태아, 분만 당시 재태 연령 37주 이상 대 미만, 출생 당시 신생아 체중 2.5 kg 이상 대 미만으로 각각 나누어, 두 군간에 농도를 비교한 결과, 통계학적으로 유의한 차이는 없었다. 본 연구는 한국인 임신부를 대상으로 최초로 양수 내 BPA 농도에 관한 보고이다. 본 연구결과 BPA 농도는 임신부 나이나 태아성별에 따라 차이를 보이지 않았으며, 조산이나 저체중아의 발생과도 관련이 없음을 알 수 있었다. 향후 산전 BPA 노출이 배발생에 영향을 미치는지에 대한 연구가 필요하다고 사료된다.

• 한국발생생물학회지:발생과생식
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• 제22권3호
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• pp.263-273
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• 2018

Aquaporin (AQP) 3, a facilitated transporter of water and glycerol, expresses in placenta and fetal membranes, but the detailed localization and function of AQP3 in placenta remain unclear. To elucidate a role of AQP3 in placenta, we defined the expression and cellular localization of AQP3 in placenta and fetal membranes, and investigated the structural and functional differences between wild-type and AQP3 null mice. Gestational sacs were removed during mid-gestational period and amniotic fluid was aspirated for measurements of volume and composition. Fetuses with attached placenta and fetal membranes were weighed and processed for histological assessment. AQP3 strongly expressed in basolateral membrane of visceral yolk sac cells of fetal membrane, the syncytiotrophoblasts of the labyrinthine placenta and fetal nucleated red blood cell membrane. Mice lacking AQP3 did not exhibit a significant defect in differentiation of trophoblast stem cells and normal placentation. However, AQP3 null fetuses were smaller than their control litter mates in spite of a decrease in litter size. The total amniotic fluid volume per gestational sac was reduced, but the amniotic fluid-to-fetal weight ratio was increased in AQP3 null mice compared with wild-type mice. Glycerol, free fatty acid and triglyceride levels in amniotic fluid of AQP3 null mice were significantly reduced, whereas lactate level increased when compared to those of wild-type mice. These results suggest a role for AQP3 in supplying nutrients from yolk sac and maternal blood to developing fetus by facilitating transport of glycerol in addition to water, and its implication for the fetal growth in utero.

• 한국발생생물학회지:발생과생식
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• 제20권1호
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• pp.63-71
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• 2016

Human embryonic stem cells (hESCs) have been routinely cultured on mouse embryonic fibroblast feeder layers with a medium containing animal materials. For clinical application of hESCs, animal-derived products from the animal feeder cells, animal substrates such as gelatin or Matrigel and animal serum are strictly to be eliminated in the culture system. In this study, we performed that SNUhES32 and H1 were cultured on human amniotic fluid cells (hAFCs) with KO-SR XenoFree and a humanized substrate. All of hESCs were relatively well propagated on hAFCs feeders with xeno-free conditions and they expressed pluripotent stem cell markers, alkaline phosphatase, SSEA-4, TRA1-60, TRA1-81, Oct-4, and Nanog like hESCs cultured on STO or human foreskin fibroblast feeders. In addition, we observed the expression of nonhuman N-glycolylneuraminic acid (Neu5GC) molecules by flow cytometry, which was xenotransplantation components of contamination in hESCs cultured on animal feeder conditions, was not detected in this xeno-free condition. In conclusion, SNUhES32 and H1 could be maintained on hAFCs for humanized culture conditions, therefore, we suggested that new xeno-free conditions for clinical grade hESCs culture will be useful data in future clinical studies.

• 약학회지
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• 제41권2호
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• pp.212-219
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• 1997

Multiple forms of extracellular phospholipase $A_2$ have been detected in human amniotic fluid (HAF). When HAF was subjected to heparin-Sepharose column chromatography, phospholipase $A_2$ activity was detected in both heparin-non binding and binding fraction. The activity of heparin-non binding fraction was further purified by sequential uses of column chromatographies on butyl-Toy-opearl 650M and DEAE-Sephacel. DEAE-Sephacel fraction contained three different phospholipase $A_2$ activities (Peak I, II, III). The molecular weight of DEAE-Sephacel fraction phospholipase $A_2$ determined by SDS-PAGE were about 52KDa (Peak I). Peak II, III required micromolar $Ca^{2+}$ ion for its maximum activity, but Peak I enzyme showed calcium independent phospholipase $A_2$ activity and showed broad range of pH (6.0~10.0) optimum. All these enzymes were not recognized by a monoclonal antibody raised against phospholipase $A_2$ from human synovial fluid. These results suggest that HAF might contain multiple forms of extracellular phospholipase $A_2$, which may neither belong to the 14KDa group II phospholipase $A_2$ family nor cytosolic phospholipase $A_2$.

• Toxicological Research
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• 제26권4호
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• pp.275-283
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• 2010

Placenta transfer study in non-human primate (NHP) is one of the crucial components in the assessment of developmental toxicity because of the similarity between NHP and humans. To establish the method to determine placenta transfer in non-human primate, toxicokinetics of valproic acid (VPA), a drug used to treat epilepsy in pregnant women, were determined in pregnant cynomolgus monkeys. After mating, pregnancy-proven females were daily administered with VPA at dose levels of 0, 20, 60 and 180 mg/kg by oral route during the organogenesis period from gestation day (GD) 20 to 50. Concentrations of VPA and its metabolite, 4-ene-VPA, in maternal plasma on GDs 20 and 50, and concentrations of VPA and 4-ene-VPA in placenta, amniotic fluid and fetus on GD 50 were analyzed using LC/MS/MS. Following single oral administration of VPA to pregnant monkeys, concentrations of VPA and 4-ene-VPA were generally quantifiable in the plasma from all treatment groups up to 4-24 hours post-dose, demonstrating that VPA was absorbed and the monkeys were systemically exposed to VPA and 4-ene-VPA. After repeated administration of VPA to the monkeys, VPA was detected in amniotic fluid, placenta and fetus from all treatment groups, demonstrating that VPA was transferred via placenta and the fetus was exposed to VPA, and the exposures were increased with increasing dose. Concentrations of 4-ene-VPA in amniotic fluid and fetus were below the limit of quantification, but small amount of 4-ene-VPA was detected in placenta. In conclusion, pregnant monkeys were exposed to VPA and 4-ene-VPA after oral administration of VPA at dose levels of 20, 60 and 180 mg/kg during the organogenesis period. VPA was transferred via placenta and the fetus was exposed to VPA with dose-dependent exposure. The metabolite, 4-ene VPA, was not detected in both amniotic fluid and fetus, but small amount of 4-ene-VPA was detected in placenta. These results demonstrated that proper procedures to investigate placenta transfer in NHP, such as mating and diagnosis of pregnancy via examining gestational sac with ultrasonography, collection of amniotic fluid, placenta and fetus after Caesarean section followed by adequate bioanalysis and toxicokinetic analysis, were established in this study using cynomolugus monkeys.

• Journal of Microbiology and Biotechnology
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• 제20권6호
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• pp.959-967
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• 2010

Down syndrome (DS) is an abnormality of the 21st chromosome that commonly occurs in children born to older women. Thus, amniotic fluid (AF) is usually collected from such women for prenatal diagnosis. This study analyzed human AF supernatants (AFS) using a mass spectrometric (MS) approach to search for candidate biomarkers of a DS pregnancy. The AFS were collected from older pregnant women at weeks 16-18 of their gestation by amniocentesis for cytogenetic analysis. The AFS from the pregnancies carrying DS (n=4) or chromosomally normal (n=6) fetuses, as revealed by the cytogenetic analysis, were then subjected to global protein profiling based on liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Affinity chromatography was also applied prior to the LC-ESI-MS/MS to minimize the masking effect of highly abundant albumin and immunoglobulin and thereby increase the diversity of the identified proteins. As a result, at least 30 new AFS proteins were identified and 44 AFS proteins were found to be differentially expressed between the DS and normal cases, where 6 of the proteins were unique to the DS cases and 11 were unique to the chromosomally normal cases. In addition, in the DS cases, 19 AFS proteins were downregulated and 8 were upregulated to varying degrees. A Western blot analysis confirmed the LC-ESI-MS/MS data, indicating that the combined detection of apolipoprotein A-II (apoA-II) and alpha-fetoprotein (AFP) could be a potential tool for diagnosing DS cases.

• 한국발생생물학회지:발생과생식
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• 제22권2호
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• pp.199-203
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• 2018

Although trisomy 16 is commonly detected in spontaneous abortions and accounts for over 30% of cases of autosomal trisomy detected after spontaneous abortion, trisomy 16 mosaicism is rarely detected by amniocentesis in the second trimester. Here, we report a case of level III trisomy 16 mosaicism (47,XX,+16[8]/46,XX[31]) diagnosed by cytogenetic analysis of independently cultured amniotic fluid cells. The female baby was delivered at full term with low birth weight and intrauterine growth retardation, and interestingly, her karyotype was normal (46,XX). Given the difficulty in predicting the outcomes of fetuses with this mosaicism, it is recommended to inform the possibility of mosaicisms including this trisomy 16 mosaicism during prenatal genetic diagnosis and genetic counseling for parents.