• 한국동물학회지
• /
• 제27권3호
• /
• pp.127-136
• /
• 1984

Xenopus 수정란을 중력에 대해 새로운 방향으로 기울이거나 연속회전시키는 일련의 방법을 통하여 배/복측 극성 (dorsal/ventral polarity) 형성에 관계되는 다음과 같은 사실을 알게 되었다. (1) 수정 후 난을 $15^\\circ, 30^\\circ, 45^\\circ 및 60^\\circ$ 각도로 기울인 채 발생시키면 원구배순부는 보통중력의 반대쪽에 형성되었으며, $15^\\circ에서 60^\\circ$로 기울이는 각도를 증가시킬수록 변경되는 율도 증가하였다. (2) 자외선을 조사한 수정란을 제 1 분열전 $15^\\circ, 30^\\circ, 45^\\circ, 60^\\circ$등의 각도를 유지한 상태에서 발생시킨 결과 자외선에 의한 축형성의 발생결함이 회복되었으며 이때 기울이는 각도가 클수록 그 회복율도 증가하였다. (3) 여러 속도의 정속회전대 (clinostat)상에서 수정란을 발생시킬 경우 비교적 저속인 $0.45 \\sim 9.0$ rph에서는 원구배순부의 형성 장소가 난의 회전방향에 의존하였으나, $40 \\sim 360$ rph에서는 무작위한 방향으로 나타났다. 한편 정속회전대를 경험한 embryo는 대부분 정상적인 발생을 보였으나 축형성상의 결함을 보이는 개체도 아울러 관찰되었다. 이상과 같은 결과들을 배/복측 극성 결정기작과 연관지어 논의하였다.

• Biomolecules & Therapeutics
• /
• 제26권4호
• /
• pp.389-398
• /
• 2018

p-Cresol, found at high concentrations in the serum of chronic kidney failure patients, is known to cause cell senescence and other complications in different parts of the body. p-Cresol is thought to mediate cytotoxic effects through the induction of autophagy response. However, toxic effects of p-cresol on mesenchymal stem cells have not been elucidated. Thus, we aimed to investigate whether p-cresol induces senescence of mesenchymal stem cells, and whether melatonin can ameliorate abnormal autophagy response caused by p-cresol. We found that p-cresol concentration-dependently reduced proliferation of mesenchymal stem cells. Pretreatment with melatonin prevented pro-senescence effects of p-cresol on mesenchymal stem cells. We found that by inducing phosphorylation of Akt and activating the Akt signaling pathway, melatonin enhanced catalase activity and thereby inhibited the accumulation of reactive oxygen species induced by p-cresol in mesenchymal stem cells, ultimately preventing abnormal activation of autophagy. Furthermore, preincubation with melatonin counteracted other pro-senescence changes caused by p-cresol, such as the increase in total 5'-AMP-activated protein kinase expression and decrease in the level of phosphorylated mechanistic target of rapamycin. Ultimately, we discovered that melatonin restored the expression of senescence marker protein 30, which is normally suppressed because of the induction of the autophagy pathway in chronic kidney failure patients by p-cresol. Our findings suggest that stem cell senescence in patients with chronic kidney failure could be potentially rescued by the administration of melatonin, which grants this hormone a novel therapeutic role.

• 해양환경안전학회지
• /
• 제18권4호
• /
• pp.331-335
• /
• 2012

2012년 1월 13일 이탈리아 국적 초대형 여객선 코스타 콘코디아호가 부분 침몰되었다. 당시 해당 선박은 4,252명의 여객과 승무원이 승선하고 있었으며, 사고 며칠 후 30여명이 사망하였으며 사고 24시간 이후 두명의 한국인을 포함한 이탈리아 선원들이 극적으로 구조되었다. 2012년 6월 18일까지(본 논문 발표일 기준) 총 32명의 사망자와 64명의 부상자 등 총 100여명의 사상자가 발생한 해사안전 문제의 치유할 수 없는 사고가 발생되었고 현재까지 사고에 대한 조사가 여전히 진행 중이다. 본 연구에서는 이번 초대형 이탈리아 여객선 코스타 콘코디아호의 재난 경과 보고서로써 사고의 문제점, 원인, 향후 대책방안 등에 관하여 조사 분석을 실시하였다. 본 연구의 하나의 주된 결과로써 향후 이러한 초대형 여객 운반선의 경우 국제 해사 안전적 관점에서 혁신적이고 지능적인 선박 퇴선 시스템이 구축되어야 할 것으로 분석되었다.

• The Korean Journal of Physiology and Pharmacology
• /
• 제20권3호
• /
• pp.279-286
• /
• 2016

Caffeic acid phenethyl ester (CAPE), derived from honeybee hives, is a bioactive compound with strong antioxidant activity. This study was designed to test the neuroprotective effect of CAPE in 3-nitropropionic acid (3NP)-induced striatal neurotoxicity, a chemical model of Huntington's disease (HD). Initially, to test CAPE's antioxidant activity, a 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) antioxidant assay was employed, and CAPE showed a strong direct radical-scavenging effect. In addition, CAPE provided protection from 3NP-induced neuronal cell death in cultured striatal neurons. Based on these observations, the in vivo therapeutic potential of CAPE in 3NP-induced HD was tested. For this purpose, male C57BL/6 mice were repeatedly given 3NP to induce HD-like pathogenesis, and 30 mg/kg of CAPE or vehicle (5% dimethyl sulfoxide and 95% peanut oil) was administered daily. CAPE did not cause changes in body weight, but it reduced mortality by 29%. In addition, compared to the vehicle-treated group, robustly reduced striatal damage was observed in the CAPE-treated animals, and the 3NP-induced behavioral deficits on the rotarod test were significantly rescued after the CAPE treatment. Furthermore, immunohistochemical data showed that immunoreactivity to glial fibrillary acidic protein (GFAP) and CD45, markers for astrocyte and microglia activation, respectively, were strikingly reduced. Combined, these data unequivocally indicate that CAPE has a strong antioxidant effect and can be used as a potential therapeutic agent against HD.

• Journal of Microbiology and Biotechnology
• /
• 제25권6호
• /
• pp.782-787
• /
• 2015

In this study, we developed an assay system for missense mutations in human phenylalanine hydroxylases (hPAHs). To demonstrate the reliability of the system, eight mutant proteins (F39L, K42I, L48S, I65T, R252Q, L255V, S349L, and R408W) were expressed in a mutant strain (pah^-) of Dictyostelium discoideum Ax2 disrupted in the indigenous gene encoding PAH. The transformed pah - cells grown in FM minimal medium were measured for growth rate and PAH activity to reveal a positive correlation between them. The protein level of hPAH was also determined by western blotting to show the impact of each mutation on protein stability and catalytic activity. The result was highly compatible with the previous ones obtained from other expression systems, suggesting that Dictyostelium is a dependable alternative to other expression systems. Furthermore, we found that both the protein level and activity of S349L and R408W, which were impaired severely in protein stability, were rescued in HL5 nutrient medium. Although the responsible component(s) remains unidentified, this unexpected finding showed an important advantage of our expression system for studying unstable proteins. As an economic and stable cell-based expression system, our development will contribute to mass-screening of pharmacological chaperones for missense PAH mutations as well as to the in-depth characterization of individual mutations.

• The Korean Journal of Physiology and Pharmacology
• /
• 제18권3호
• /
• pp.249-254
• /
• 2014

The purpose of this study is to characterize the effects of KHG26792 (3-(naphthalen-2-yl(propoxy)methyl)azetidine hydrochloride), a potential skin whitening agent, on melanin synthesis and identify the underlying mechanism of action. Our data showed that KHG26792 significantly reduced melanin synthesis in a dose-dependent manner. Additionally, KHG26792 downregulated microphthalmia-associated transcription factor (MITF) and tyrosinase, the rate-limiting enzyme in melanogenesis, although tyrosinase was not inhibited directly. KHG26792 activated extracellular signal-regulated kinase (ERK), whereas an ERK pathway inhibitor, PD98059, rescued KHG26792-induced hypopigmentation. These results suggest that KHG26792 decreases melanin production via ERK activation. Moreover, the hypopigmentary effects of KHG26792 were confirmed in a pigmented skin equivalent model using Cervi cornus Colla (deer antler glue), in which the color of the pigmented artificial skin became lighter after treatment with KHG26792. In summary, our findings suggest that KHG26792 is a novel skin whitening agent.

• 대한수의학회지
• /
• 제57권2호
• /
• pp.97-104
• /
• 2017

Solanum lycopersicum, commonly known as tomato, is widely used in raw, cooked, or liquid forms because it contains nutritional compounds that are beneficial for human health, including carotenoids, lycopene, ascorbic acid, vitamins, and minerals. The tomato is perhaps the most widely studied fruit, especially with respect to its cardioprotective effects. In this study, we aimed to identify the anti-inflammatory mechanisms by which the tomato elicits its anti-inflammatory properties. We treated murine macrophage RAW 264.7 cells with a tomato ethanol extract and performed various biochemical assays including nitric oxide inhibition, cell viability, RNA extraction, expression of pro-inflammatory mediators and cytokines, and immunoblotting, as well we assessed cell survival rates. Our results have shown for the first time that a tomato ethanol extract treatment can suppress nitric oxide production in a dose-dependent manner without cytotoxicity. Moreover, it inhibits the expression of pro-inflammatory mediators and cytokines and elicits its anti-inflammatory effects via the nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$) and mitogen-activated protein kinase (MAPK) pathways. In addition, administration of tomato syrup potently rescued mice from septic shock induced by lipopolysaccharide injection. Collectively, our results elucidate details regarding the anti-inflammatory mechanisms of tomato.

• IMMUNE NETWORK
• /
• 제3권4호
• /
• pp.268-275
• /
• 2003

Background: Hemopoietic cells require the constant presence of growth factors for survival in vitro and in vivo. Caspases have been known as central executors of apoptotic cell death. We have, therefore, investigated the pathways that regulate caspase activity and apoptosis using the $CD34^+$ cell line, TF-1 which requires GM-CSF for survival. Methods: Apoptosis was measured by annexin V staining and mitochondrial membrane potential was measured by DiOC6 labelling. Intracellular pH was measured using pH sensitive fluorochrome, BCECF or SNARF-1, followed by flow cytometry analysis. Caspase activation was analyzed by PARP cleavage using anti-PARP antibody. Results: Removal of GM-CSF induceed PARP cleavage, a hallmark of caspase activity, concomitant with pHi acidification and a drop in mitochondrial potential. Treatment with ZVAD, a competitive inhibitor of caspases, partially rescued cell death without affecting pHi acidification and the reduction of mitochondrial potential, suggesting that both these events act upstream of caspases. Overexpression of Bcl-2 prevented cell death induced by GM-CSF deprivation as well as pHi acidification and the reduction in mitochondrial membrane potential. In parental cells maintained with GM-CSF, EIPA, a competitive inhibitor of $Na^+/H^+$ antiporter induced apoptosis, accompanied by a drastic reduction in mitochondrial potential. In contrast, EIPA induced apoptosis in Bcl-2 transfectants without causing mitochondrial membrane depolarization. Conclusion: Taken together, our results suggest that the regulation of $H^+$fluxes, either through a mitochondriondependent or independent pathway, is central to caspase activation and apoptosis.

• Molecules and Cells
• /
• 제37권2호
• /
• pp.126-132
• /
• 2014

As a tumor suppressor homologue during mitosis, Chk2 is involved in replication checkpoints, DNA repair, and cell cycle arrest, although its functions during mouse oocyte meiosis and early embryo development remain uncertain. We investigated the functions of Chk2 during mouse oocyte maturation and early embryo development. Chk2 exhibited a dynamic localization pattern; Chk2 expression was restricted to germinal vesicles at the germinal vesicle (GV) stage, was associated with centromeres at pro-metaphase I (Pro-MI), and localized to spindle poles at metaphase I (MI). Disrupting Chk2 activity resulted in cell cycle progression defects. First, inhibitor-treated oocytes were arrested at the GV stage and failed to undergo germinal vesicle breakdown (GVBD); this could be rescued after Chk2 inhibition release. Second, Chk2 inhibition after oocyte GVBD caused MI arrest. Third, the first cleavage of early embryo development was disrupted by Chk2 inhibition. Additionally, in inhibitor-treated oocytes, checkpoint protein Bub3 expression was consistently localized at centromeres at the MI stage, which indicated that the spindle assembly checkpoint (SAC) was activated. Moreover, disrupting Chk2 activity in oocytes caused severe chromosome misalignments and spindle disruption. In inhibitor-treated oocytes, centrosome protein ${\gamma}$-tubulin and Polo-like kinase 1 (Plk1) were dissociated from spindle poles. These results indicated that Chk2 regulated cell cycle progression and spindle assembly during mouse oocyte maturation and early embryo development.

• Molecules and Cells
• /
• 제40권9호
• /
• pp.643-654
• /
• 2017

Autophagy is a degradation pathway in eukaryotic cells in which aging proteins and organelles are sequestered into double-membrane vesicles, termed autophagosomes, which fuse with vacuoles to hydrolyze cargo. The key step in autophagy is the formation of autophagosomes, which requires different kinds of vesicles, including COPII vesicles and Atg9-containing vesicles, to transport lipid double-membranes to the phagophore assembly site (PAS). In yeast, the cis-Golgi localized t-SNARE protein Sed5 plays a role in endoplasmic reticulum (ER)-Golgi and intra-Golgi vesicular transport. We report that during autophagy, sed5-1 mutant cells could not properly transport Atg8 to the PAS, resulting in multiple Atg8 dots being dispersed into the cytoplasm. Some dots were trapped in the Golgi apparatus. Sed5 regulates the anterograde trafficking of Atg9-containing vesicles to the PAS by participating in the localization of Atg23 and Atg27 to the Golgi apparatus. Furthermore, we found that overexpression of SFT1 or SFT2 (suppressor of sed5 ts) rescued the autophagy defects in sed5-1 mutant cells. Our data suggest that Sed5 plays a novel role in autophagy, by regulating the formation of Atg9-containing vesicles in the Golgi apparatus, and the genetic interaction between Sft1/2 and Sed5 is essential for autophagy.