• 대한소아치과학회지
• /
• 제28권4호
• /
• pp.709-727
• /
• 2001

세포자기사(programmed cell death)는 배자발생과정에서 중요한 역할을 하는 정상적인 생리적현상으로 알려져왔다. 생쥐 배자에서 세포자기사에 관한 많은 연구가 있었지만 초기 형태발생기동안의 전반적이고 구체적인 세포자기사 양상에 관한 보고는 없었다. 이에 본 연구에서는 발생 4.5일$\sim$11.5일째의 Balb/c생쥐배자에서 생쥐발생초기의 세포자기사가 일어나는 양상을 알아보았다. 세포자기사가 일어나는 양상은 배자 조직절편과 온전한 배자에 in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) 반응을 시켜 알아보았다. 주머니포배시기 (발생 4.5일째)에는 세포자기사 과정에 있는 세포가 극히 드물게 속세포명이에서 관찰되었다. Egg cylinder시기 초기(발생 $5.0\sim5.5$일째)에는 소수의 세포자기사 세포가 배자외배엽과 배자내배엽, 원시양막공간에서 나타났다. Egg cylinder시기 후반부(발생 $5.5\sim6.5$일)에는 세포자기사 세포가 배자외배엽, 배자내배엽, 원시양막공간 뿐만 아니라 바깥배자외배엽과 바깥배자내배엽 에서도 관찰되었다. Streak 시기 (발생 $6.75\sim7.75$일)에는 많은 세포자기사 세포가 ectoplacental cone부위에서 관찰된 반면에, 배자바깥 부위에서는 융모막과 바깥배자내배엽에서 매우 적은 수의 세포자기사 세포가 관찰되었고, 배자부위에서는 소수의 세포자기사 세포가 무작위적으로 배자외배엽과 중배엽, 그리고 배자내배엽 부위에서 관찰되었다. 체절형성기 초기 (발생 $8.0\sim8.5$일)에는 많은 수의 세포자기사 세포가 신경주름(neural fold)의 가장 앞쪽(머리쪽)에 주로 분포하는 것이 관찰되었다. 체절형성기 중기(발생 $9.0\sim9.5$일)에 귀기원판(otic placode)부위에서 처음으로 세포자기사 세포가 관찰되었고, 적은 수의 세포자기사 세포들이 또한 눈기원판(optic placode)과 아가미궁에서도 관찰되기 시작하였다. 발생 $9.5\sim9.75$일째에 크게 세가지 흐름의 세포자기사 세포들이 머리쪽 부위에서 관찰되었다. 발생 10.5일째에는 세포자기사 세포들이 발생중인 눈 부위와 안쪽코돌기와 바깥쪽코돌기 그리고 상악돌기가 만나는 부위, 아가미궁의 가쪽부위, 양쪽 하악돌기가 만나는 중앙부위, 그리고 팔다리싹의 꼭대기외배엽능선에 국한되어 관찰되었다. 발생 11.5일째에는 세포자기사 세포가 발생중인 팔다리부위와 꼬리부위를 제외한 나머지 부위에서는 현저히 감소되어 나타났다. 본 연구에서는 생쥐 발생초기 동안에 나타나는 세포자기사 세포가 발생시기에 따라 어느 부위에 발현되는 가는 알아보았는데, 이러한 연구 결과는 배자의 형태발생을 연구하는데 기초적인 자료로 활용될 수 있을 것으로 사료된다.

• Radiation Oncology Journal
• /
• 제21권4호
• /
• pp.315-321
• /
• 2003

목적: 발육 중인 백서 태아 대뇌 피질에서 방사선조사에 의한 아포토시스 반응에 대하여 알아보고자 하였다 대상 및 방법: 임신 백서의 태령 17 내지 19일(E17$\~$E19)에 선형가속기의 X-ray를 이용하여 어미 쥐의 복부에 방사선을 조사하였다. 방사선량에 따른 아포토시스를 보기 위하여 1, 2, 3, 4 Gy를 전후 조사면으로 방사선조사를 시행하고 5시간 후 백서 태아의 뇌를 획득하였다. 시간 경과에 따른 아포토시스를 보기 위하여 E7$\~$E19에 2 Gy의 방사선을 조사한 후 1, 3, 6, 12, 24시간에 각각 백서 태아의 뇌를 획득하였다. 대조군은 방사선을 조사하지 않은 임신 쥐를 같은 조건하에서 사육하여 각 군당 3마리씩 할당하였다. 아포토시스 세포는 면역조직화학적 방법(TUNEL, In situ 707-mediated dUTP nick end labeling)으로 염색하여 관찰하였다 결과: 백서 태아 대뇌 피질에서 TUNEL 양성인 세포는 광학현미경하에서 전형적인 아포토시스의 형태학적 특징을 나타내었다 대조군에서는 대뇌 피질의 전 층에서 TUNEL 양성세포를 거의 발견할 수 없었다. 1 Gy의 방사선 조사 후 5시간에 대뇌피질 전 층에서 미약하게 관찰되었으나 2 Gy에는 전 층에서 1 Gy 경우보다 더 증가하였고 그중 뇌실대와 중간대에서 피질대보다 더 많이 나타났다. 조사된 1$\~$4 Gy 범위에서 방사선량이 증가할수록 TUNEL양성세포가 더욱 증가하는 것을 볼 수 있었다. 2 Gy의 방사선 조사 후 3시간부터 TUNEL양성세포가 관찰 되기 시작하여 방사선 조사 후 6시간에 최고점을 이루었으며 이는 24시간까지 지속되었다. 결론: 발육 중인 백서 태아 대뇌피질에서 방사선에 의한 아포토시스의 전형적인 형태학적 특징을 관찰할 수 있었다. 아포토시스는 뇌실대와 중간대에서 피질대보다 더 많이 나타났으며 이는 줄기세포와 초기의 분화세포에서 방사선조사에 대한 감수성이 더욱 민감함을 시사하였다.

• Journal of Korean Neurosurgical Society
• /
• 제63권6호
• /
• pp.821-826
• /
• 2020

Objective : Hyperostosis in meningiomas can be present in 4.5% to 44% of cases. Radical resection should include aggressive removal of invaded bone. It is not clear however to what extent bone removal should be carried to achieve pathologically free margins, especially that in many cases, there is a T2 hyperintense signal that extends beyond the hyperostotic bone. In this study we try to investigate the perimeter of tumour cells outside the visible nidus of hyperostotic bone and to what extent they are present outside this nidus. This would serve as an initial step for setting guidelines on dealing with hyperostosis in meningioma surgery. Methods : This is a prospective case series that included 14 patients with convexity meningiomas and hyperostosis during the period from March 2017 to August 2018 in two university hospitals. Patients demographics, clinical, imaging characteristics, intraoperative and postoperative data were collected and analysed. In all cases, all visible abnormal bone was excised bearing in mind to also include the hyperintense diploe in magnetic resonance imaging (MRI) T2 weighted images after careful preoperative assessment. To examine bony tumour invasion, five marked bone biopsies were taken from the craniotomy flap for histopathological examinations. These include one from the centre of hyperostotic nidus and the other four from the corners at a 2-cm distance from the margin of the nidus. Results : Our study included five males (35.7%) and nine females (64.3%) with a mean age of 43.75 years (33-55). Tumor site was parietal in seven cases (50%), fronto-parietal in three cases (21.4%), parieto-occipital in two cases (14.2%), frontal region in one case and bicoronal (midline) in one case. Tumour pathology revealed a World Health Organization (WHO) grade I in seven cases (50%), atypical meningioma (WHO II) in five cases (35.7%) and anaplastic meningioma (WHO III) in two cases (14.2%). In all grade I and II meningiomas, bone biopsies harvested from the nidus revealed infiltration with tumour cells while all other bone biopsies from the four corners (2 cm from nidus) were free. In cases of anaplastic meningiomas, all five biopsies were positive for tumour cells. Conclusion : Removal of the gross epicentre of hyperostotic bone with the surrounding 2 cm is adequate to ensure radical excision and free bone margins in grade I and II meningiomas. Hyperintense signal change in MRI T2 weighted images, even beyond visible hypersototic areas, doesn't necessarily represent tumour invasion.

• Animal cells and systems
• /
• 제6권3호
• /
• pp.227-232
• /
• 2002

GroESLx proteins of symbiotic X-bacteria were overproduced in Escherichia coli and their structural characteristics were assayed after simple purification. The GroESx and GroELx were heat-stable at 8$0^{\circ}C$ and 5$0^{\circ}C$, respectively. After heat-treatment, GroESx was purified by DEAE Sephadex A-50 chromatography and GroELx was purified by step- and linear sucrose density gradient ultracentrifugation. Molecular masses of GroESx and GroELx were 50-80 kDa and 800 kDa, respectively, as estimated by sucrose density gradient ultracentrifugation. In chemical cross-linking analysis, subunits of GroESx were mostly cross-linked by incubation for 3 h in 0.4% glutaralde-hyde and GroESx was found to be composed of homo-heptamer subunits. Those of GroELx were cross-linked within 10 min in 0.3% glutaraldehyde and GroELx was in two stacks of homo-heptamer subunits. On the other hand, GroESx and GroELx proteins in a solution could not be cross-linked even after incubation for 3 h in 0.5% glutaraldehyde. GroELx was stable at 4-37$^{\circ}C$. In the presence of both GroESx and ATP, GroELx$_{14}$ was stable at 37$^{\circ}C$ but not at 4$^{\circ}C$ or 24$^{\circ}C$. Thus, we confirmed the oligomeric properties of GroESx$_{7}$ and GroELx$_{14}$ and their stability to heat and in the interaction with GroESx.x.

• The Korean Journal of Physiology and Pharmacology
• /
• 제25권4호
• /
• pp.341-354
• /
• 2021

Cardamonin (CARD) is a chalconoid with anti-inflammatory and antioxidant properties, and it is present in several plants. We sought to explore whether CARD exerts any positive effects against hyperglycemia-induced testicular dysfunction caused by type 2 diabetes and aimed to identify its possible intracellular pathways. Adult male rats were subdivided into six groups: control, CARD, diabetic (DM), DM + glibenclamide (GLIB), DM + CARD and DM + GLIB + CARD. Type 2 DM induced a significant increase in blood glucose and insulin resistance, along with diminished serum insulin, testosterone and gonadotropins levels, which were associated with the impairment of key testicular androgenic enzymes and cellular redox balance. Administration of CARD at a dose of 80 mg/kg for 4 weeks effectively normalized all of these alterations, and the improvement was confirmed by epididymal sperm analysis. After treatment with CARD, the pathological changes in spermatogenic tubules were markedly improved. Significantly, CARD upregulated testicular glucose transporter-8 (GLUT-8) expression and had inhibitory effects on elevated autophagy markers and caspase-3 immunoreactive cells. Furthermore, our results revealed that CARD was able to attenuate damage via activation of Nrf2 through the p62-dependent degradation of testicular anti-Kelch-like ECH-associated protein-1 (Keap-1). In conclusion, this study suggests that CARD provides protection against diabetic stress-mediated testicular damage. The use of CARD with conventional anti-diabetic therapy was associated with improved efficacy compared with conventional therapy alone.

• 대한치과교정학회지
• /
• 제30권2호
• /
• pp.205-214
• /
• 2000

Nd-Fe-B 자석의 정자기장 (Static Magnetic Field)이 MC3T3-E1 세포의 alkaline phosphase (ALP) 활성도에 미치는 영향을 알아보기 위하여, MC3T3-El세포를 12 well 세포배양접시의 1열과 3열에 접종하고 1열과 3열의 첫째 칸 하방에 Nd-Fe-B 자석을 가하여 7일, 13일, 19일, 25일간 배양한 후 세포의 ALP 활성도를 측정하였으며, 100 mm 세포배양접시의 한 쪽 가장자리 하방에 Nd-Fe-B 자석을 가하여 7일, 13일, 19일, 25일간 배양한 후 ALP 염색을 하여 다음과 같은 결과를 얻었다. 1. 정자기장을 가한 13, 19, 25일에 각각 100 mT에서는 대조군에 비해 ALP활성도가 감소된 반면 4.6 mT와 0.5 mT에서는 대조군에 비해 ALP 활성도가 증가되었다 (P<0.01). 2. ALP염색에서 전체적으로는 19일까지 ALP가 증가되었다가 25일에 다소 감소되는 양상을 나타내었으며, 각 세포배양접시에서는 7일에는 뚜렷한 차이를 보이지 않았으나 13, 19, 25일에는 자석이 놓인 부위 (100 mT) 가 그 반대편(0.5 mT)에 비해 ALP가 감소되었음을 육안으로나 도립위상차현미경으로 관찰할 수 있었다. 이상의 결과 0.5 mIT와 4.6 mT의 낮은 자기장에서는 ALP 활성도가 증가되었으며 100 mT의 높은 자기장에서는 ALP활성도가 감소되어, Nd-Fe-B자석의 정자기장이 MC3T3-E1 세포의 ALP 활성도에 영향을 미쳐 골조직의 형성 및 개조에 영향을 미칠 수 있을 것으로 생각된다.

• Journal of Microbiology and Biotechnology
• /
• 제28권5호
• /
• pp.739-747
• /
• 2018

The low solubility and high-cost recovery of Propionibacterium acnes polyunsaturated fatty acid isomerase (PAI) are key problems in the bioproduction of high value-added conjugated linoleic acid (CLA). To improve the solubility of recombinant PAI, six chaperone proteins were coexpressed with PAI. Introduction of GroELS proteins dramatically improved the PAI solubility from 29% to 97%, with increased activity by 57.8%. Combined expression of DnaKJ-GrpE and GroELS proteins increased the activity by 11.9%. In contrast, coexpression of DnaKJ-GrpE proteins significantly reduced the activity by 57.4%. Plasmids pTf16 harboring the tig gene and pG-Tf2 containing the tig and groEL-groES genes had no visible impact on PAI expression. The lytic protein E was then introduced into the recombinant Escherichia coli to develop a cell autolysis system. A 35% activity of total intracellular PAI was released from the cytoplasm by suspending the lysed cells in distilled water. The PAI recovery was further improved to 81% by optimizing the release conditions. The lipase from Rhizopus oryzae was also expressed in E. coli, with an extracellular activity of 110.9 U/ml. By using the free PAI and lipase as catalysts, a joint system was established for producing CLA from sunflower oil. Under the optimized conditions, the maximum titer of t-10,c-12-CLA reached 9.4 g/l. This work provides an effective and low-cost strategy to improve the solubility and recovery of the recombinant intracellular PAI for further large-scale production of CLA.

• 치위생과학회지
• /
• 제21권4호
• /
• pp.243-250
• /
• 2021

Background: Endodontic sealers or their toxic components may become inflamed and lead to delayed wound healing when in direct contact with periapical tissues over an extended period. Moreover, an overfilled sealer can directly interact with adjacent tissues and may cause immediate necrosis or further resorption. Therefore, the treatment outcome conceivably depends on the endodontic sealer's biocompatibility and osteogenic potential. This study aimed to evaluate the cell viability and osteogenic effects of four different sealers in osteoblastic cells. Methods: AH Plus (resin-based sealer), Pulp Canal Sealer EWT (zinc oxide-eugenol sealer), BioRoot RCS (calcium silicate-based sealer), and Well-Root ST (MTA-based calcium silicate sealer) were mixed strictly according to the manufacturer's instructions, and dilutions of sealer extracts (1/2, 1/5 and 1/10) were determined. Cell viability was measured using the water-soluble tetrazolium-8 (WST-8) assay. Differentiation was assessed by alkaline phosphatase (ALP) activity and mineralized nodule formation by Alizarin Red S staining. Results: The cell viability of the extracts derived from the sealers excluding Well-Root ST was concentration dependent, with sealer extracts having the least viability at a 1/2 dilution. At sealer extract dilution of 1/10, the test groups showed the same survival rate as that control group, with the exception of BioRoot RCS. Among all experimental groups, BioRoot RCS showed the highest cell viability after 48 hours. The ALP activity was significantly higher in a concentration-dependent manner. Furthemore, all four materials promoted ALP activity and mineralized nodule formation compared to the control at 1/10 dilutions. Conclusion: This is the first study to highlight the differences in biological activity of these four materials. These results suggest that the composition of root canal sealers appears to alter the form of biocompatibility and osteoblastic differentiation.

• Parasites, Hosts and Diseases
• /
• 제52권6호
• /
• pp.613-619
• /
• 2014

Neospora caninum (Apicomplexa; Sarcocystidae) is a protozoan that causes abortion in cattle, horses, sheep, and dogs as well as neurological and dermatological diseases in dogs. In the central nervous system of dogs infected with N. caninum, cysts were detected that exhibited gliosis and meningitis. Flavonoids are polyphenolic compounds that exhibit antibacterial, antiparasitic, antifungal, and antiviral properties. In this study, we investigated the effects of flavonoids in a well-established in vitro model of N. caninum infection in glial cell cultures. Glial cells were treated individually with 10 different flavonoids, and a subset of cultures was also infected with the NC-1 strain of N. caninum. All of the flavonoids tested induced an increase in the metabolism of glial cells and many of them increased nitrite levels in cultures infected with NC-1 compared to controls and uninfected cultures. Among the flavonoids tested, 3',4'-dihydroxyflavone, 3',4',5,7-tetrahydroxyflavone (luteolin), and 3,3',4',5,6-pentahydroxyflavone (quercetin), also inhibited parasitophorous vacuole formation. Taken together, our findings show that flavonoids modulate glial cell responses, increase NO secretion, and interfere with N. caninum infection and proliferation.

• Asian-Australasian Journal of Animal Sciences
• /
• 제24권11호
• /
• pp.1514-1524
• /
• 2011

A total of 4,752 monthly lactation records of Friesian cows during the period from 2000 to 2005 were used to estimate genetic parameters and to determine the effect of udder health on milk production traits. Three milk production traits were studied: 305-day milk yield (305-dMY), 305-day fat yield (305-dFY) and 305-day protein yield (305-dPY). Four udder health traits were studied: somatic cell count (SCC), mastitis (MAST), udder health status (UDHS) with 10 categories and udder quarter infection (UDQI) with 7 categories. Mixed model least square analysis was used to estimate the fixed effects of month and year of calving and parity (P) on different studied traits. Sire and dam within sire were included in the model as random effects. Data were analyzed using Multi-trait Derivative Free Restricted Maximum Likelihood methodology (MTDFREML) to estimate genetic parameters. Unadjusted means of 305-dMY, 305-dFY, 305-dPY and SCC were 3,936, 121, 90 kg and 453,000 cells/ml, respectively. Increasing SCC from 300,000 to 2,000,000 cells/ml increased UDQI from 5.51 to 23.2%. Losses in monthly and lactationally milk yields per cow ranged from 17 to 93 and from 135 to 991 kg, respectively. The corresponding losses in monthly and lactationally milk yields return per cow at the same level of SCC ranged from 29.8 to 163 and from 236 to 1,734 Egyptian pounds, respectively. Heritability estimates of 305-dMY, 305-dFY, 305-dPY, SCC, MAST, UDHS, UDQI were 0.31${\pm}$0.4, 0.33${\pm}$0.03, 0.35${\pm}$0.05, 0.23${\pm}$0.02, 0.14${\pm}$0.02, 0.13${\pm}$0.03, and 0.09${\pm}$0.01, respectively. All milk production traits showed slightly unfavorable negative phenotypic and genetic correlations with SCC, MAST, UDHS and UDQI. There were positive and high genetic correlations between SCC and each of MAST (0.85${\pm}$0.7), UDHS (0.87${\pm}$0.10) and UDQI (0.77${\pm}$0.06) and between MAST and each of UDHS (0.91${\pm}$0.11) and UDQI (0.83${\pm}$0.07). It could be concluded that the economic losses from mastitis and high SCC are considerable. The high genetic correlation between SCC and clinical mastitis (CM) suggest that the selection for lower SCC would help to reduce or eliminate the undesirable correlated responses of clinical mastitis associated with selection for increasing milk yield. Additionally, it is recommended also that if direct information on under health traits is not available, measures of SCC can be inclusion in a selection criteria to improve the income from dairy cows.