• 한국발생생물학회지:발생과생식
• /
• 제21권2호
• /
• pp.121-130
• /
• 2017

4-Nonylphenol (NP) is a surfactant that is a well-known and widespread estrogenic endocrine disrupting chemical (EDC). Although it has been known that the affinity of NP to ERs is low, it has been suggested that low-dose NP has toxicity. In the present study, the endocrine disrupting effects on reproduction, and the weight of gonads, epididymis, and uterus were evaluated with the chronic lower-dose NP exposing. This study was designed by following the OECD test guideline 443 and subjected to a complete necropsy. In male, NP had an effect on the weight of the testis and epididymis in both $F_0$ and $F_1$. In females, NP decreased the weight of ovary and uterus in $F_0$ but not in pre-pubertal $F_1$ pubs. Fertility of male and female in $F_0$ or $F_1$ was no related with NP administration. The number of caudal-epididymal sperm by body weight (BW) was not different between groups in both $F_0$ and $F_1$. Besides, the difference of the sperm number between generations was not detected. The number of ovulated oocytes was similar between groups in $F_0$, but significantly decreased in NP 50 group of $F_1$. The litter size and sex ratios of offspring in $F_1$ and $F_2$ were not different. The accumulated mating rate and gestation period were not affected by the NP administration. Those results shows that chronic lower-dose NP administration has an effect of endocrine disruptor on the weight of gonads and epididymis of $F_0$ and $F_1$ but not in reproduction. Based on the results, it is suggested that chronic lower-dose NP exposing causes endocrine disruption in the weight of gonad and epididymis but not in the reproductive ability of next generations.

• BMB Reports
• /
• 제35권3호
• /
• pp.313-319
• /
• 2002

N-Acetyl-$\beta$-D-hexosaminidase ($\beta$-HexNAc'ase) (EC 3.2.1.52) was purified from rice seeds (Oryza sative L. var. Dongjin) using ammonium sulfate (80%) precipitation, Sephadex G-150, CM-Sephadex, and DEAE-Sephadex chromatography, sequentially. The activities were separated into 7 fractions($F_1-F_7$) by CM-Sephadex chromatography. Among them, F6 was further purified to homogeneity with a 13.0% yield and 123.3 purification-fold. The molecular mass was estimated to be about 52 kDa on SDS-PAGE and 37.4 kDa on Sephacryl S-300 gel filtration. The enzyme catalyzed the hydrolysis of both p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GlcNAc) and p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GalNAc) as substrates, which are typical properties of $\beta$-HexNAc'ase. The ratio of the pNP-GlcNAc'ase activity to the pNP-GalNAc'ase activity was 4.0. However, it could not hydrolyze chitin, chitosan, pNP-$\beta$-glucopyranoside, or pNP-$\beta$-glucopyranoside. The enzyme showed $K_m$, $V_{max}$ and $K_{cat}$ for pNP-GlcNAc of 1.65 mM, $79.49\;mM\;min^{-1}$, and $4.79{\times}10^6\;min^{-1}$, respectively. The comparison of kinetic values for pNP-GlcNAc and pNP-GalNAc revealed that the two enzyme activities are associated with a single binding site. The purified enzyme exhibited optimum pH and temperature for pNP-GlcNAc of 5.0 and $50^{\circ}C$, respectively. The enzyme activity for pNP-GlcNAc was stable at pH 5.0-5.5 and $20-40^{\circ}C$. The enzyme activity was completely inhibited at a concentration of 0.1 mM $HgCl_$ and $AgNO_3$, suggesting that the intact thiol group is essential for activity. Chloramine T completely inhibited the activity, indicating the possible involvement of methionines in the mechanism of the enzyme.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• 제33권3호
• /
• pp.191-198
• /
• 2007

The p53 which is well known as tumor suppressor gene is located at 17p13. p53 is a sequence-specific DNA binding transcription factor that responds to certain cytotoxic stresses, such as DNA damage, by enhancing the transcription of genes that regulate cell-cycle progression as well as programmed cell death. The p63 gene that is located at 3q27-29, is recognized members of the p53 family, and responsible for the transcription of 6 isoforms. Three isoforms ($TAp63{\alpha}$, $TAp63{\beta}$, $TAp63{\gamma}$) contain an N-terminal transactivation (TA) domain and can induce apoptosis. The other 3 isoforms (${\Delta}Np63{\alpha}$, ${\Delta}Np63{\beta}$, ${\Delta}Np63{\gamma}$) lack the TA domain and may function in a dominant-negative fashion by inhibiting the transactivation functions of p53 and TAp63 proteins, and thus act as oncoproteins. A number of studies have investigated the role of p63 in human squamous cell carcinomas from different organs. Only a few studies have examined ${\Delta}Np63$ isoform in oral squamous cell carcinoma including normal epithelium. This study aimed to evaluate expression of ${\Delta}Np63$ isoform in human oral squamous cell carcinoma tissue and normal mucosa. The 3 cases of well differenciated oral squamous cell carcinoma specimen including adjacent normal mucosa were examined, and immunohistochemical study with monoclonal antibody(4A4) and tumor cell apoptosis analysis with Transmission Electon Microscopy were studied. And, RT-PCR analysis was done for expression of ${\Delta}Np63$ isoform. The results were as followed. 1. Normal gingiva showed the restricted p63 expression in basal cell layer. 2. Well differentiated squamous cell carcinoma showed mainly p63 expression in overall area of malignancy, especially in basal cell layer to adjacent stromal tissue. 3. Tumor cells around keratinized area with no p63 expression disclosed less micro-organelle in decreased size cytoplasm and severe chromatin margination with nuclear destruction that means apoptosis. 4. Comparison of mRNA expression of ${\Delta}Np63$ isoform by RT-PCR showed variable expression of ${\Delta}Np63$ isoform, but ${\Delta}Np63{\alpha}$ was most highly expressed in all 3 tumor specimen. From theses results, it should be suggested that ${\Delta}Np63$ isoform expression in well differentiated squamous cell carcinoma was closely related to tumor oncogenesis, expecially overexpression of ${\Delta}Np63{\alpha}$ is a most important factor in tumor genesis of oral squamous cell carcinoma.

• Environmental Analysis Health and Toxicology
• /
• 제20권3호
• /
• pp.195-203
• /
• 2005

Cytochrome P45O 17$\alpha$-hydroxylase (CYPI 7) and cytorhrome P45O aromata.ie (CYPI 9) are key steroidogenic enzymes in androgen and estrogen synthesis. ThiL study evaluated the effects of nonylphenol (NP) on CYP17 and CYP19 expression in the ovary of Sprague-Dawley rats. All female rats were administered orally with the vehicle (control, corn oil), diethylstilbestrol (DES, 5.0 $\mu$g/kg) and NP (50, 100, or 200 mg/kg/day), which was startinB when they were weaned at 21 days of age for 20 days. Twenty four hours after final dose, the animals were anelthetized with ether. Significant decreases in the uterus (wet weight) were observed with 5.0 $\mu$g/kg/day DES (78$\%$, of control) and 200 mg/kg/day NP (62$\%$ of control), respectively Additionally, ovarian weight was significantly decreased with 5.0 $\mu$g/kg/day DES (63$\%$ of control) and 200 mg/kg/day NP (72$\%$ of control). The serum estradiol levels were sligHtly lower in DES and high dose NP treatment groups, but the 74 levels were not affected by DES and NP. The expression of the ovarian CYP19 gene increased with low doses (50 and 100 mg/kg/day) of NP. while DES and high dose oi NP (200 mg/kg/day) did not affect on the CYP19 mRNA levels. In contrast to the CYP19 gene, the CYP17 gene expreLsion level was significantly down-regulated by the DES and 200 mg/ks/day NP. This result suggestE that NP inhibits ovarian estrogen synthelis by supprelsing CYP17 mRNA efprelsion, And different mechanisml might exist for the expression of Lteroidogenic CYP17 and CYP19 genes in the ovary of Sprague-Dawley rats in response to NP.

• Biomolecules & Therapeutics
• /
• 제17권1호
• /
• pp.92-97
• /
• 2009

As nanomaterials might enter into cells and have high reactivity with intracellular structures, it is necessary to assay possible genotoxic risk of them. One of these approaches, we investigated possible genotoxic potential of gold nanoparticle (AuNP) using L5178Y cells. Four different sizes of AuNP (4, 15, 100 or 200 nm) were synthesized and the sizes and structures of AuNP were analyzed using transmission electron microscopy (TEM), scanning electron microscopy (SEM) and stability was analyzed by a UV/Vis. Spectrophotometer. Cytotoxicity was assessed by direct cell counting, and cellular location was detected by dark field microscope at 6, 24 and 48 h after treatment of AuNP. Comet assay was conducted to examine DNA damage and tumor necrosis factor (TNF)-${\alpha}$ mRNA level was assay by real-time reverse transcription polymerase chain reaction. Synthetic AuNP (4, 50, 100 and 200 nm size) had constant characteristics and stability confirmed by TEM, SEM and spectrophotometer for 10 days, respectively. Dark field microscope revealed the location of AuNP in the cytoplasm at 6, 24 and 48 h. Treatment of 4 nm AuNP induced dose and time dependent cytotoxicity, while other sizes of AuNP did not. However, Comet assay represented that treatment of 100 nm and 200 nm AuNP significantly increased DNA damage compared to vehicle control (p <0.01). Treatment of 100 nm and 200 nm AuNP significantly increased TNF-${\alpha}$ mRNA expression compared to vehicle control (p<0.05, p<0.01, respectively). Taken together, AuNP induced DNA damage in L5178Y cell, associated with induction of oxidative stress.

• 대한핵의학회지
• /
• 제31권1호
• /
• pp.108-115
• /
• 1997

NP-59 신티그라피는 부신피질의 기능장애를 선택적으로 영상화할 수 있는 장점이 있기에 다양한 부신질환의 진단에 적응이 되는바 특히 임상검사와 방사선학적 검사 결과가 서로 상이한 경우 진단에 결정적인 도움을 줄 수 있는 기능 검사법이다. 저자는 이러한 장점을 가지고 있는 NP-59 스캔을 부신질환 환자에서 시행하여 그 결과를 임상자료와 비교하여 부신피질 신티그라피의 유용성을 규명해보고자 이 연구를 시행하였다. 본 연구대상으로서 1990년 3월부터 1996년 12월까지 동아대학교병원 핵의학과에서 NP-59 스캔을 시행한 10명의 환자와 그들의 스캔 11예를 대상으로 하여 병력지에 기재된 임상자료와 스캔판독소견을 후향적으로 비교 분석하였다. 대상환자 중 쿠싱병(2예)과 부신종양에 의한 쿠싱증후군(3예)에서는 진단명과 스캔판독소견이 일치하였다. 부신의 부수종양(2예)에서는 1예는 진단명과 판독소견이 일치하였으나 1예에 있어서는 기능성 종양으로 판독되어 불일치를 보였는데 이는 subclinical Cushin-g's syndrome으로 추정되어 관찰중이다. 임상적으로 는 일차성 알도스테론증과 부신성 조모증이 의심되나 부신 CT 검사상 정상소견을 보인 환자에서 이를 확인하기 위하여 덱사메타존 억제스캔을 시행한 결과 정상소견을 보였고 이들은 각각 본태성 고혈압과 간경화증에 동반된 조모증으로 추정되어 치료 및 경과 관찰중이다. 이상의 결과로 보아 NP-59 신티그라피는 쿠싱증후군의 진단과 원인질환의 감별진단에 매우 유용하였으며 비록 대상수가 작아서 결론 내리기는 어려우나 부수종양에서 스캔소견만으로도 전이성 병변을 제외시킬 수 있으므로 결과적으로는 불필요한 생검을 피할 수 있게 하는 임상적인 유용성을 가지고 있었다.

• 한국유기농업학회지
• /
• 제23권1호
• /
• pp.91-101
• /
• 2015

'추황배' 배 과수원에 봉지종류와 수확시기를 달리하였을 때 수상의 과피흑변과 발생과 수확 후 과실품질에 어떠한 영향을 미치는 지를 규명하기 위하여 수행되었다. 본 시험은 두 가지로, 첫 번째는 네 종류의 봉지(신문지외지/황색내지, 회색외지/황색내지, 신문지외지/흑색내지, 신문지외지/적색내지) 시험이었고, 두 번째 시험은 만개 후 160일, 170일, 180일, 190일, 200일에 수확하였을 때 과피흑변과와 과실품질을 조사하였다. 신문지외지/황색내지의 봉지가 투광도와 투기도 및 수분투과도인 물리성이 향상되었고, 신문지외지/적색내지가 봉지물리성이 낮은 경향을 보였다. 신문지외지/황색내지는 수상에서의 과피흑변과 발생이 나타나지 않았고, 신문지외지/적색내지를 괘대처리 한 과실에서는 약 12.7%의 과피흑변과가 관찰되었다. 과피흑변은 주로 과실의 중앙부와 과정부에서 집중적으로 발생되었다. 신문지외지/적색내지를 이용한 유대재배는 평균과중, 경도, 산도, 그리고 착색이 향상되는 경향을 보였다. 수확시기가 늦어질수록(만개 후 190일과 200일에 수확) 과피흑변, 평균과중, 당도가 증가되었고 반대로 과실경도와 산도는 감소되었다.

• Journal of Microbiology and Biotechnology
• /
• 제25권11호
• /
• pp.1827-1834
• /
• 2015

The SMG1 lipase from Malassezia globosa is a newly found mono- and diacylglycerol (DAG) lipase that has a unique lid in the loop conformation that differs from the common alpha-helix lid. In the present study, we characterized the contribution of three residues, L103 and F104 in the lid and F278 in the rim of the binding site groove, on the function of SMG1 lipase. Site-directed mutagenesis was conducted at these sites, and each of the mutants was expressed in the yeast Pichia pastoris, purified, and characterized for their activity toward DAG and p-nitrophenol (pNP) ester. Compared with wild-type SMG1, F278A retained approximately 78% of its activity toward DAG, but only 11% activity toward pNP octanoate (pNP-C8). L103G increased its activity on pNP-C8 by approximately 2-fold, whereas F104G showed an approximate 40% decrease in pNP-C8 activity, and they both showed decreased activity on the DAG emulsion. The deletion of 103-104 retained approximately 30% of its activity toward the DAG emulsion, with an almost complete loss of pNP-C8 activity. The deletion of 103-104 showed a weaker penetration ability to a soybean phosphocholine monolayer than wild-type SMG1. Based on the modulation of the specificity and activity observed, a pNP-C8 binding model for the ester (pNP-C8, N102, and F278 form a flexible bridge) and a specific lipid-anchoring mechanism for DAG (L103 and F104 serve as "anchors" to the lipid interface) were proposed.

• 한국환경과학회지
• /
• 제20권2호
• /
• pp.231-239
• /
• 2011

Diethyl phthalate (DEP) and nonylphenol (NP) are widely spread in the natural environment as an endocrine disruption chemicals (EDs). Therefore, in this study, ultrasound (US) and ultraviolet (UVC), including $TiO_2$, as advanced oxidation processes (AOPs) were applied to a DEP and NP contaminated solution. When only the application of US, the optimum frequency for significant DEP degradation and a high rate of hydrogen peroxide ($H_2O_2$) formation was 283 kHz. We know that the main mechanism of DEP degradation is radical reaction and, NP can be affected by both of radical reaction and pyrolysis through only US (sonolysis) process and combined US+UVC (sonophotolysis) process. At combined AOPs (sonophotolysis/sonophotocatalysis) such as US+UVC and US+UVC+$TiO_2$, significant degradation of DEP and NP were observed. Enhancement effect of sonophotolysis and sonophotocatalysis system of DEP and NP were 1.68/1.38 and 0.99/1.17, respectively. From these results, combined sonophotocatalytic process could be more efficient system to obtain a significant DEP and NP degradation.

• 공업화학
• /
• 제7권1호
• /
• pp.18-25
• /
• 1996

교반소요동력은 장치의 설계상 중요할 뿐만 아니라 교반 프로세스를 평가, 검토하는 데 매우 중요하다. 특히 실험실 레벨에서 플랜트로의 스케일업은 주로 동력기준으로 행해지기 때문에 장치형상의 선택과 동력의 설정이 교반효과를 결정한다. 따라서 본 연구에서는 난류역 완전방해판 조건하에서 경사 임펠러 및 2단 패들 임펠러의 적절한 동력상관식을 제시한다. $$Np_{(pitch)}=({\theta}/90^{\circ})Np_{(90^{\circ})}$$ 또한 임펠러 사이의 거리에 따른 동력수의 추산식을 제시한다. $$Np=7.09(n_p)^{0.7}(\frac{b_(double)}{d})(\frac{H}{D})^{0.18}$$, $${n_p}^{0.7}(\frac{b_{(double)}}d)$$<2 $$Np=8.73\{(n_p)^{0.7}(\frac{b_{(double)}}{d})\}^{0.7}(\frac{H}{D})^{0.18}$$, $${n_p}^{0.7}(\frac{b_{(double)}}d)$$>2.