• 한국동물생명공학회지
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• 제36권3호
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• pp.145-153
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• 2021

The role of transient receptor potential vanilloid receptor-1 (TRPV1) has been primarily investigated in pain sensory neurons. Relatively, little research has been performed in testicular cells. TRPV1 is abundantly expressed in Leydig cells of young adult mice. This study was conducted to determine the role of the TRPV1 channel in Leydig cells. TRPV1 modulators and testosterone were treated to the mouse Leydig cell line TM3 cells for 24 h. Capsaicin, a TRPV1 activator, dose-dependently induced cell death, whereas capsazepine, a TRPV1 inhibitor, inhibited capsaicin-induced cell death. Testosterone treatment reduced capsaicin-induced cell death. High concentrations of testosterone decreased TRPV1 mRNA and protein expression levels. However, TRPV1 modulators did not affect testosterone production. These results showed that capsaicin induced cell death of Leydig cells and that testosterone reduced capsaicin-induced cell death. Our findings suggest that testosterone may regulate the survival of Leydig cells in young adult mice by decreasing the expression level of TRPV1.

• Korean Journal of Acupuncture
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• 제20권1호
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• pp.57-64
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• 2003

목적: 오가피가 mouse testis에서 유래된 Leydig 세포주에서 에탄올에 의해 유발된 아폽토시스에 미치는 영향을 조사하였다. 방법: TM3 세포주에서의 아폽토시스 변화를 관찰하기 위해서 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI), DNA fragmentation assay, 및 reverse transcription-polymerase chain reaction (RT-PCR) 방법을 이용하였다. 결과: MTT assay를 이용하여 분석한 결과 농도에 따른 세포독성의 효과가 에탄올 투여부터 관찰되었다. 또한 오가피로 전처치하고 에탄올을 처치하였을 때 세포독성이 크게 감소되었다. DAPI staining에서 오가피 투여군은 에탄올 투여군에 비해서 fragmentation이 억제되었다. RT-PCR의 분석에 의하여 caspase-3 mRNA 발현이 오가피 투여군은 알코올 투여군보다 유의성있게 억제됨을 보여주었다. 결론: TM3 Leydig 세포주에서 에탄올에 의해 유발된 아폽토시스는 전형적인 세포사멸 형태를 나타내었다. 반면에 오가피 투여군은 에탄올에 의해서 유발된 아폽토시스에서 세포보호 효과가 있음이 확인되었다.

• Molecules and Cells
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• 제27권4호
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• pp.391-396
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• 2009

Image-based, high-content screening assays demand solutions for image segmentation and cellular compartment encoding to track critical events - for example those reported by GFP fusions within mitosis, signalling pathways and protein translocations. To meet this need, a series of nuclear/cytoplasmic discriminating probes have been developed: DRAQ5$^{TM}$ and CyTRAK Orange$^{TM}$. These are spectrally compatible with GFP reporters offering new solutions in imaging and cytometry. At their most fundamental they provide a convenient fluorescent emission signature which is spectrally separated from the commonly used reporter proteins (e.g. eGFP, YFP, mRFP) and fluorescent tags such as Alexafluor 488, fluorescein and Cy2. Additionally, they do not excite in the UV and thus avoid the complications of compound UV-autofluorescence in drug discovery whilst limiting the impact of background sample autofluorescence. They provide a convenient means of stoichiometrically labelling cell nuclei in live cells without the aid of DMSO and can equally be used for fixed cells. Further developments have permitted the simultaneous and differential labelling of both nuclear and cytoplasmic compartments in live and fixed cells to clearly render the precise location of cell boundaries which may be beneficial for quantitative expression measurements, cell-cell interactions and most recently compound in vitro toxicology testing.

• Molecules and Cells
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• 제38권12호
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• pp.1037-1043
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• 2015

The TAZ activator 2-butyl-5-methyl-6-(pyridine-3-yl)-3-[2'-(1H-tetrazole-5-yl)-biphenyl-4-ylmethyl]-3H-imidazo[4,5-b]pyridine] (TM-25659) inhibits adipocyte differentiation by interacting with peroxisome proliferator-activated receptor gamma. 1 TM-25659 was previously shown to decrease weight gain in a high fat (HF) diet-induced obesity (DIO) mouse model. However, the fundamental mechanisms underlying the effects of TM-25659 remain unknown. Therefore, we investigated the effects of TM-25659 on skeletal muscle functions in C2 myotubes and C57BL/6J mice. We studied the molecular mechanisms underlying the contribution of TM-25659 to palmitate (PA)-induced insulin resistance in C2 myotubes. TM-25659 improved PA-induced insulin resistance and inflammation in C2 myotubes. In addition, TM-25659 increased FGF21 mRNA expression, protein levels, and FGF21 secretion in C2 myotubes via activation of GCN2 pathways (GCN2-$phosphoelF2{\alpha}$-ATF4 and FGF21). This beneficial effect of TM-25659 was diminished by FGF21 siRNA. C57BL/6J mice were fed a HF diet for 30 weeks. The HF-diet group was randomly divided into two groups for the next 14 days: the HF-diet and HF-diet + TM-25659 groups. The HF diet + TM-25659-treated mice showed improvements in their fasting blood glucose levels, insulin sensitivity, insulin-stimulated Akt phosphorylation, and inflammation, but neither body weight nor food intake was affected. The HF diet + TM-25659-treated mice also exhibited increased expression of both FGF21 mRNA and protein. These data indicate that TM-25659 may be beneficial for treating insulin resistance by inducing FGF21 in models of PA-induced insulin resistance and HF diet-induced insulin resistance.

• 대한본초학회지
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• 제26권1호
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• pp.133-138
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• 2011

Objectives : The purpose of this study was to estimate the antioxidant activity of Sesami Semen Nigrum extract (SSN) on mouse Leydig cells, TM3. Methods : Cell viability assays were performed. The protective effects of SSN against hydrogen peroxide-induced oxidative stress in Leydig cells were examined by measuring cell viability. Lipid peroxidation levels and antioxidant enzyme concentrations such as SOD and catalase were measured. Results : Cell viability of Leydig cells increased with SSN concentration. Cell viability of Leydig cells was 136.66% when SSN concentration was $50{\mu}g/ml$. Cell viability of the hydrogen peroxide group was statistically decreased (p<0.01) compared with the control group. Antioxidant effect of SSN was measured and the protective effect of SSN concentration were 5, 10, $50{\mu}g/ml$. LPO were decreased significantly at 5, $50{\mu}g/ml$ of SSN concentrations. SOD activity was increased at 1, 10, $50{\mu}g/ml$ of SSN concentrations. Catalase activity was significantly increased at 123.7, 133.3 and 131.9 units/mg protein when SSN concentrations were 5, 10 and $50{\mu}g/ml$, respectively. Conclusions : In conclusion, Sesami Semen Nigrum extract has antioxidant activities in Leydig cells against oxidative stress.

• 한국안광학회지
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• 제12권2호
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• pp.67-78
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• 2007

시판중인 렌즈 보존액 중 가장 흔히 사용 또는 유통되고 있는 CLIWELL$^{TM}$에 대한 안점막자극성 시험을 식품의약품 안전청 고시에 따라 Draize법을 이용하여 그 자극성의 정도를 평가하고, 눈에 가해질 수 있는 잠재적인 독성을 평가하기 위하여, 배출물에 대한 도말표본 상에서 염증세포 및 비 염증성 유상피세포가 차지하는 비율과 함께 점안 14일 후 안구 중, 각막, 홍채, 망막 및 공막의 조직병리학적 관찰을 실시한 결과 다음과 같은 결과를 얻었다. 비 세척군에서는 점안 후 1, 2 및 3일에 국한하여, 각막과 결막에 경미한 자극이 관찰되어, 이 기간 동안의 평균 안점막 자극지수 MIOI가 각각 4.50, 1.67 및 0.67로 관찰되었으며, 세척군에서는 점안 후 1 및 2일에 국한하여, 결막에서 경미한 자극성이 인정되었고, 이때 MIOI는 각각 0.67로 관찰되었다. 따라서 CLIWELL$^{TM}$은 세척군 및 비 세척군 모두에서 MIOI가 모든 관찰 기간에서 4.50이하로 관찰되었으며, IAOI 역시 점안 1일 후 4.50으로 관찰되어 무자극성 물질로 판단되었다. 한편 배출물에 대한 염증성 세포의 비율은 무처치 대조군과 비교하여 점안 1일 후 유의성 있는 염증세포 비율의 증가가 비세척군에 국한하여 인정된 이외에 의미 있는 변화는 인정되지 않았다. 또한 안구의 조직병리학적 검사에서 각막, 홍채, 망막 및 공막 모두에서 무처치 대조군과 비교하여 의미 있는 변화는 인정되지 않았다.

• 한국전자파학회논문지
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• 제21권8호
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• pp.886-892
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• 2010

본 논문에서는 epsilon negative(ENG) 영차 공진과 $TM_{010}$-모드를 결합하여 넓은 대역폭을 갖고, 지향성 방사 패턴을 갖는 하이브리드 안테나를 제안한다. ENG 영차 공진을 얻기 위해 버섯 구조를 이용하였고, $TM_{010}$ 모드를 얻기 위해 패치 안테나를 이용하였다. 이 두 개의 안테나 사이에 갭을 주어 하나의 동축 선로 급전 구조로 두 개의 안테나를 결합하였다. ENG ZOR 주파수는 4 GHz, $TM_{010}$의 주파수는 3.9 GHz로 설계하였다. 두 주파수가 근접해 있기 때문에 이중 공진을 형성할 수 있고, 따라서 대역폭을 확장시킬 수가 있다. ENG ZOR 안테나는 전방향성의 방사 패턴을 갖지만, ENG ZOR 주파수와 $TM_{010}$의 주파수가 매우 근접해 있기 때문에 전방향성의 방사 패턴의 널 부분을 $TM_{010}$ 모드 방사가 보상해줄 수 있어 제안된 안테나는 지향성 방사 패턴을 갖는다. 제안된 안테나는 버섯 구조의 단위 셀을 각각 4개, 3개, 2개로 줄여나가면서 3가지 종류로 설계하여 특성을 분석하였다. 각각의 제안된 안테나는 비대역폭 $4.29{\sim}4.95 %$, 이득 3.16~5.57 dBi, 효율 62.4~94.2 %의 특성을 가지고 있다.

• BMB Reports
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• 제52권11호
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• pp.665-670
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• 2019

Nucleotide-binding oligomerization domain protein 2 (NOD2), an intracellular pattern recognition receptor, plays important roles in inflammation and cell death. Previously, we have shown that NOD2 is expressed in vascular smooth muscle cells (VSMCs) and that NOD2 deficiency promotes VSMC proliferation, migration, and neointimal formation after vascular injury. However, its role in endoplasmic reticulum (ER) stress-induced cell death in VSMCs remains unclear. Thus, the objective of this study was to evaluate ER stress-induced viability of mouse primary VSMCs. NOD2 deficiency increased ER stress-induced cell death and expression levels of apoptosis mediators (cleaved caspase-3, Bax, and Bak) in VSMCs in the presence of tunicamycin (TM), an ER stress inducer. In contrast, ER stress-induced cell death and expression levels of apoptosis mediators (cleaved caspase-3, Bax, and Bak) were decreased in NOD2-overexpressed VSMCs. We found that the $IRE-1{\alpha}-XBP1$ pathway, one of unfolded protein response branches, was decreased in NOD2-deficient VSMCs and reversed in NOD2-overexpressed VSMCs in the presence of TM. Furthermore, NOD2 deficiency reduced the expression of XBP1 target genes such as GRP78, PDI-1, and Herpud1, thus improving cell survival. Taken together, these data suggest that the induction of ER stress through NOD2 expression can protect against TM-induced cell death in VSMCs. These results may contribute to a new paradigm in vascular homeostasis.

• Restorative Dentistry and Endodontics
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• 제23권1호
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• pp.366-378
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• 1998

Thermocondensation root canal filling technique have been used to fill accessary canals or to obtain homogeneous root caral fillings. But these thermocondensation technique inevitably produce heat in the canal which can be transmitted through the dentin and cementum and consequently damage periodontal ligamental cells and osteoblasts. In this study, System $B^{TM}$(Analytic technology, WA.D.S.A.) was used to evaluate the reaction of periodontal ligament tissue to "Continous Wave condensation technique" introduced by Buchanan, and the transmitted root surface temperature was measured according to measured root thicknesses. 12 Mandibular incisors of two adult dogs were used for the experiment. 6 controls were filled by lateral condensation technique with sealer.3 specimens were apically filled by Continuous Wave technique at $200^{\circ}C$ for 5 seconds and remaining 3 specimens were additionally backfilled using System $B^{TM}$(Analytic technology, WA.D.S.A.) was used to evaluate the reaction of periodontal ligament tissue to "Continous Wave condensation technique" introduced by Buchanan, and the transmitted root surface temperature was measured according to measured root thicknesses. 12 Mandibular incisors of two adult dogs were used for the experiment. 6 controls were filled by lateral condensation technique with sealer.3 specimens were apically filled by Continuous Wave technique at $200^{\circ}C$ for 5 seconds and remaining 3 specimens were additionally backfilled using System $B^{TM}$ at $100^{\circ}C$ for 20 seconds. Six weeks later, the dogs were sacrificed and the teeth stained with Hematoxylin and Eosin for histologic examination. 6 extracted human teeth were used to measure the transmitted temperature. After cutting off the crown, the canals were prepared and divided into 3 groups with root thickness of 1.5mm, 1.0mm, 0.5mm, 2 teeth in each group. Inside each root canal, System $B^{TM}$ was heated as with the temperature for the apically condensed and the back filled group, and the transmitted heat was measured on the external surface of the root. The temperature of System $B^{TM}$ heat spreader at $200^{\circ}C$ and $100^{\circ}C$ was also measured at root temperature. It can be concluded as follows: 1. In the thin area (200-$250{\mu}m$) of the root, root resorption could be seen even with heating at $200^{\circ}C$ for 5 seconds. 2. When the spreader was heated at $200^{\circ}C$ for 5 seconds and additionally at $100^{\circ}C$ for 20 seconds for backfill, all teeth showed root resorption regardless of their root thickness. 3. The transmitted external root surface temperature was higher as the root thickness decreased and as the heating time increased. In the thermocompaction technique using System $B^{TM}$, the spreader should be heated for the minimal time and used only in the apical area. The heated spreader shouldn't inserted to the binding point of the canal and backfilling should be done with other means of minimally heated gutta percha technique.

• 대한의생명과학회지
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• 제16권3호
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• pp.193-196
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• 2010

Multiplex PCR-based short tandem repeat (STR) analysis is considered as a good tool for monitoring bone marrow engraftment after sex-mismatched allogeneic transplantation and provides a sensitive and accurate assessment of the contribution of both donor and/or recipient cells in post-transplantation specimens. Forensic STR analysis and quantitative real time PCR are used to determine the proportion of donor versus recipient each contained within the total DNA. The STR markers were co-amplified in a single reaction by using commercial $PowerPlex^{(R)}$ 16 system and $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ / $Yfiler^{(R)}$ PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI $PRIS^{(R)}$ 3100 Genetic Analyzer with capillary electrophoresis. The $GeneMapper^{TM}$ ID software were used for size calling and analysis of STR profiles. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA / Y Human Male DNA Quantification Kit The intent of this study was to analyze the ratio of donor versus recipient cells in the post-transplant peripheral blood, spleen, lung and kidney specimens. Specimens were taken from the traffic accident male victim who had been engrafted from bone marrow female donor. Blood and spleen specimens displayed female donor DNA profile. Kidney specimen showed male recipient DNA profile. Interestingly, lung tissue showed mixed profiles. The findings of this study indicate that the forensic STR analysis using fluorescence labeling PCR combined with capillary electrophoresis is quick and reliable enough to assess the ratio of donor versus recipient cells and to monitor the mixed chimeric patterns.