• Title/Summary/Keyword: Fluorescence effect

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Sinus augmentation with poly(ε) caprolactone-β tricalcium phosphate scaffolds, mesenchymal stem cells and platelet rich plasma for one-stage dental implantation in minipigs

  • Jeong-Hun Nam;Akram Abdo Almansoori;Oh-Jun Kwon;Young-Kwon Seo;Bongju Kim;Young-Kyun Kim;Jong-Ho Lee;KangMi Pang
    • Journal of Periodontal and Implant Science
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    • v.53 no.3
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    • pp.218-232
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    • 2023
  • Purpose: This study evaluated the efficacy of a tube-shaped poly(ε) caprolactone - β tricalcium phosphate (PCL-TCP) scaffold with the incorporation of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) and platelet-rich plasma (PRP) for bone regeneration in the procedure of single-stage sinus augmentation and dental implantation in minipigs. Methods: Implants were placed in the bilateral sides of the maxillary sinuses of 5 minipigs and allocated to a PCL-TCP+hUCMSCs+PRP group (n=5), a PCL-TCP+PRP group (n=5), and a PCL-TCP-only group (n=6). After 12 weeks, bone regeneration was evaluated with soft X-rays, micro-computed tomography, fluorescence microscopy, and histomorphometric analysis. Results: Four implants failed (2 each in the PCL-TCP+hUCMSCs+PRP and PCLTCP+hUCMSC groups). An analysis of the grayscale levels and bone-implant contact ratio showed significantly higher mean values in the PCL-TCP+hUCMSCs+PRP than in the PCL-TCP group (P=0.045 and P=0.016, respectively). In fluoromicroscopic images, new bone formation around the outer surfaces of the scaffolds was observed in the PCLTCP+hUCMSCs+PRP group, suggesting a tenting effect of the specially designed scaffolds. Bone regeneration at the scaffold-implant interfaces was observed in all 3 groups. Conclusions: Using a tube-shaped, honeycombed PCL-TCP scaffold with hUCMSCs and PRP may serve to enhance bone formation and dental implants' osseointegration in the procedure of simultaneous sinus lifting and dental implantation.

The Effect of Photosynthesis, Stomatal Conductivity, Thermotolerance and Growth on Foliar Fertilization of Carbonated Water at Lettuce Hydroponic Cultivation (상추 양액재배 시 탄산수 엽면시비가 광합성, 기공전도, 내서성, 생육에 미치는 영향)

  • Woo, Y.H.;Kim, D.E.;Lee, J.W.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.21 no.1
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    • pp.115-122
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    • 2019
  • Foliar fertilization of carbonated water during lettuce hydroponic cultivation was increased photosynthetic rate and stomatal conductance as higher carbon dioxide concentration of carbonated water The higher the carbon dioxide concentration in the carbonated water was better growth of lettuce. However, the carbon dioxide concentration of 500 ppm and 700 ppm in the carbonated water was increased the tip-burn occurrence, and the yield was higher in the 300 ppm. the carbon dioxide concentration of 300 ppm in the carbonated water was lower in the fresh weight but increased yield resulted in the lower of the tip-burn occurrence The high temperature limits for growth were 32℃ in the control, 33℃ in the 300ppm and 34℃ in the 500 ppm according to analyze chlorophyll fluorescent Fo. The high temperature tolerance in lettuce increased approximately 4℃ by foliar fertilization treatments of carbonated water under this experiment conditions. Also the activity of SOD(superoxide dismutase), the antioxidant enzyme, was higher with high carbon dioxide concentration of the carbonated water.

Ethanol Extract of Smilax glabra Induces Apoptotic Cell Death in Human YD10B Oral Squamous Cell Carcinoma Cells

  • Young Sun Hwang
    • Journal of dental hygiene science
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    • v.23 no.3
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    • pp.216-224
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    • 2023
  • Background: Smilax glabra has various pharmacological activities and is widely used as a herbal medicine. Although the incidence of oral cancer is low, the recurrence rate is high, and the 5-year survival rate is poor. It is necessary to search for anticancer drugs that increase the effect of cancer chemotherapy on heterogeneous oral tissues and reduce the side effects on normal cells. This study aimed to investigate the effects and mechanism of ethanol extract of Smilax glabra (EESG) as an anticancer drug for oral cancer. Methods: Smilax glabra root components extracted with 70% ethanol were used to analyze their effects on cancer cells. A 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay was performed for cytotoxicity analysis. Flow cytometry was performed to determine the cell cycle phase distribution. To observe apoptotic cells, terminal deoxynucleotidyl transferase dUTP nick end labeling and γH2AX were detected by fluorescence microscope. The protein levels of cleaved PARP and caspase were analyzed using western blotting. The activation of procaspase-3 was confirmed by measuring caspase-3 activity. Results: EESG was no cytotoxic to normal gingival fibroblast but was high in YD10B oral squamous cell carcinoma (OSCC) cells. EESG treatment increased the subdiploid DNA content of YD10B cells by assessing DNA content distribution. Chromatin condensation and DNA strand breaks increased in YD10B cells treated with EESG. EESG-treated YD10B cells had high Annexin V and low propidium iodide levels, confirming that early apoptosis was induced. In addition, increased levels of γH2AX foci, a marker of DNA damage, were observed in the nuclei of EESG-treated YD10B cells. The EESG-treated YD10B cells also exhibited decreased procaspase-3 and procaspase-9 levels, increased PARP cleavage and caspase-3 activity. Conclusion: These results indicate that EESG inhibited cancer cell proliferation by inducing apoptosis in YD10B OSCC cells.

A rare ginsenoside compound K (CK) induces apoptosis for breast cancer cells

  • Seun Eui Kim;Myoung-Hoon Lee;Hye-Myoung Jang;Wan-Taek Im;Joontaik Lee;Sang-Hwan Kim;Gwang Joo Jeon
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.3
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    • pp.167-176
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    • 2023
  • Background: A breast cancer is the second leading cause of cancer death in women worldwide and among different types of breast cancers, triple-negative breast cancer (TNBC) has a poor prognosis. Methods: We investigated the potential of ginsenoside compound K (CK), an active ingredient in the bio-transformed ginsenoside, to be used as a therapeutic ingredient by examining the effects of CK on cell proliferation, apoptosis, and cancer-related gene expressions in breast cancer cells. Results: From the results of treating MCF-7, an ER and PR-positive breast cancer cells, and MDA-MB-231 (TNBC) with CK at a concentration of 0-100 µM, the half maximal inhibitory concentration (IC50) values for each cell were 52.17 µM and 29.88 µM, respectively. And also, it was confirmed that cell migration was inhibited above the IC50 concentration. In addition, fluorescence analysis of Apoptosis/Necrosis showed that CK induced apoptosis rather than necrosis of breast cancer cells. Through qPCR, it was confirmed that the expression of genes related to apoptosis and cell cycle arrest was increased in CK-treated breast cancer cells, and it acted more effectively on TNBC. However, the expression of genes related to tumor invasion and metastasis is also increased, so it is necessary to consider the timing of application of CK as a potential therapeutic anticancer compound. Conclusions: CK showed a stronger inhibitory effect in TNBC with poor prognosis but considering the high tumor invasion and metastasis-related gene expression, the timing of application of CK should be considered.

Ginsenoside F2 Restrains Hepatic Steatosis and Inflammation by Altering the Binding Affinity of Liver X Receptor Coregulators

  • Kyurae Kim;Myung-Ho Kim;Ji In Kang;Jong-In Baek;Byeong-Min Jeon;Ho Min Kim;Sun-Chang Kim;Won-Il Jeong
    • Journal of Ginseng Research
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    • v.48 no.1
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    • pp.89-97
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    • 2024
  • Background: Ginsenoside F2 (GF2), the protopanaxadiol-type constituent in Panax ginseng, has been reported to attenuate metabolic dysfunction-associated steatotic liver disease (MASLD). However, the mechanism of action is not fully understood. Here, this study investigates the molecular mechanism by which GF2 regulates MASLD progression through liver X receptor (LXR). Methods: To demonstrate the effect of GF2 on LXR activity, computational modeling of protein-ligand binding, Time-resolved fluorescence resonance energy transfer (TR-FRET) assay for LXR cofactor recruitment, and luciferase reporter assay were performed. LXR agonist T0901317 was used for LXR activation in hepatocytes and macrophages. MASLD was induced by high-fat diet (HFD) feeding with or without GF2 administration in WT and LXRα-/- mice. Results: Computational modeling showed that GF2 had a high affinity with LXRα. LXRE-luciferase reporter assay with amino acid substitution at the predicted ligand binding site revealed that the S264 residue of LXRα was the crucial interaction site of GF2. TR-FRET assay demonstrated that GF2 suppressed LXRα activity by favoring the binding of corepressors to LXRα while inhibiting the accessibility of coactivators. In vitro, GF2 treatments reduced T0901317-induced fat accumulation and pro-inflammatory cytokine expression in hepatocytes and macrophages, respectively. Consistently, GF2 administration ameliorated hepatic steatohepatitis and improved glucose or insulin tolerance in WT but not in LXRα-/- mice. Conclusion: GF2 alters the binding affinities of LXRα coregulators, thereby interrupting hepatic steatosis and inflammation in macrophages. Therefore, we propose that GF2 might be a potential therapeutic agent for the intervention in patients with MASLD.

Ginseng root-derived exosome-like nanoparticles protect skin from UV irradiation and oxidative stress by suppressing activator protein-1 signaling and limiting the generation of reactive oxygen species

  • Wooram Choi;Jeong Hun Cho;Sang Hee Park;Dong Seon Kim;Hwa Pyoung Lee;Donghyun Kim;Hyun Soo Kim;Ji Hye Kim;Jae Youl Cho
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.211-219
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    • 2024
  • Background: Recently, plant-derived exosome-like nanoparticles (PDENs) have been isolated, and active research was focusing on understanding their properties and functions. In this study, the characteristics and molecular properties of ginseng root-derived exosome-like nanoparticles (GrDENs) were examined in terms of skin protection. Methods: HPLC-MS protocols were used to analyze the ginsenoside contents in GrDENs. To investigate the beneficial effect of GrDENs on skin, HaCaT cells were pre-treated with GrDENs (0-2 × 109 particles/mL), and followed by UVB irradiation or H2O2 exposure. In addition, the antioxidant activity of GrDENs was measured using a fluorescence microscope or flow cytometry. Finally, molecular mechanisms were examined with immunoblotting analysis. Results: GrDENs contained detectable levels of ginsenosides (Re, Rg1, Rb1, Rf, Rg2 (S), Gyp17, Rd, C-Mc1, C-O, and F2). In UVB-irradiated HaCaT cells, GrDENs protected cells from death and reduced ROS production. GrDENs downregulated the mRNA expression of proapoptotic genes, including BAX, caspase-1, -3, -6, -7, and -8 and the ratio of cleaved caspase-8, -9, and -3 in a dose-dependent manner. In addition, GrDENs reduced the mRNA levels of aging-related genes (MMP2 and 3), proinflammatory genes (COX-2 and IL-6), and cellular senescence biomarker p21, possibly by suppressing activator protein-1 signaling. Conclusions: This study demonstrates the protective effects of GrDENs against skin damage caused by UV and oxidative stress, providing new insights into beneficial uses of ginseng. In particular, our results suggest GrDENs as a potential active ingredient in cosmeceuticals to promote skin health.

Reversine induces cell cycle arrest and apoptosis via upregulation of the Fas and DR5 signaling pathways in human colorectal cancer cells

  • YOUNG-LAN PARK;SANG-YOON HA;SUN-YOUNG PARK;JUNG-HO CHOI;MIN-WOO JUNG;DAE-SEONG MYUNG;HYUN-SOO KIM;YOUNG-EUN JOO
    • International Journal of Oncology
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    • v.54 no.5
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    • pp.1875-1883
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    • 2019
  • Reversine, a 2,6-diamino-substituted purine analogue, has been reported to be effective in tumor suppression via induction of cell growth arrest and apoptosis of cancer cells. However, it remains unclear whether reversine exerts anticancer effects on human colorectal cancer cells. In the present study, in vitro experiments were conducted to investigate the anticancer properties of reversine in human colorectal cancer cells. The effect of reversine on human colorectal cancer cell lines, SW480 and HCT-116, was examined using a WST-1 cell viability assay, fluorescence microscopy, flow cytometry, DNA fragmentation, small interfering RNA (siRNA) and western blotting. Reversine treatment demonstrated cytotoxic activity in human colorectal cancer cells. It also induced apoptosis by activating poly(ADP-ribose) polymerase, caspase-3, -7 and -8, and increasing the levels of the pro-apoptotic protein second mitochondria-derived activator of caspase/direct inhibitor of apoptosis-binding protein with low pI. The pan-caspase inhibitor Z-VAD-FMK attenuated these reversine-induced apoptotic effects on human colorectal cancer cells. Additionally, reversine treatment induced cell cycle arrest in the subG1 and G2/M phases via increase in levels of p21, p27 and p57, and decrease in cyclin D1 levels. The expression of Fas and death receptor 5 (DR5) signaling proteins in SW480 and HCT116 cells was upregulated by reversine treatment. Reversine-induced apoptosis and cell cycle arrest were suppressed by inhibition of Fas and DR5 expression via siRNA. In conclusion, Reversine treatment suppressed tumor progression by the inhibition of cell proliferation, induction of cell cycle arrest and induction of apoptosis via upregulation of the Fas and DR5 signaling pathways in human colorectal cancer cells. The present study indicated that reversine may be used as a novel anticancer agent in human colorectal cancer.

Effect of Exogenous Application of Salicylic Acid or Nitric Oxide on Chilling Tolerance and Disease Resistant in Pepper Seedlings (외생 살리실산과 일산화질소 처리가 고추묘의 저온 내성 및 병 저항성에 미치는 영향)

  • Park, Song-Yi;Kim, Heung-Tae;Oh, Myung-Min
    • Journal of Bio-Environment Control
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    • v.23 no.4
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    • pp.329-336
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    • 2014
  • As an abiotic stress, chilling stress is one of the major factors limiting plant growth and increasing susceptibility to pathogens. Therefore, enhancing stress tolerance in plants is an important strategy for their survival under unfavorable environmental conditions. The objective of this study was to determine the effects of the exogenous application of salicylic acid (SA) or nitric oxide (NO) on chilling tolerance in pepper seedlings. Pepper (Capsicum annuum L. 'kidaemanbal') seedlings were grown under normal growing conditions ($20/25^{\circ}C$, 15 hours photoperiod, $145{\pm}5{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, fluorescence lamps) for 23 days after transplanting. The solution (3 mL) of 1 mM SA and 0.3 mM NO with surfactant triton 0.1% were sprayed two times a week, respectively. Right after the completion of chemical application, seedlings were subjected to chilling condition at $4^{\circ}C$ for 6 hours under dark condition and then the seedlings were recovered at the normal growing conditions for 2 days. In order to assess plant tolerance against chilling stress, growth characteristics, chlorophyll fluorescence (Fv/Fm), and membrane permeability were determined after chilling stress imposition. Total phenolic concentration and antioxidant capacity were measured during the whole experimental period. Disease incidence for pepper bacterial spot and wilt was also analyzed. Pepper seedlings treated with SA or NO were maintained similar dry mass ratio, while the value in control increased caused by chilling stress suggesting relatively more water loss in control plants. Electrolyte leakage of pepper seedlings treated with SA or NO was lower than that of control 2 days after chilling treatment. Fv/Fm rapidly decreased after chilling stress in control while the value of SA or NO was maintained about 0.8. SA increased higher total phenolic concentration and antioxidant capacity than NO and control during chemical treatment. In addition, increase in total phenolic concentration was observed after chilling stress in control and NO treatment. SA had an effect on the reduction of bacterial wilt in pepper seedlings. The results from this study revealed that pre-treatment with SA or NO using foliar spray was effective in chilling tolerance and the reduction of disease incidence in pepper seedlings.

Effects of Hairy Vetch and Rye Cover on Weed Occurrences and Minor Cereal Growth (피복작물처리에 따른 잡곡의 생육과 잡초발생)

  • Jeon, Seung-Ho;Lee, Se-Hun;Oh, Se-Yun;Kim, Young-Ju;Kim, Kyung-Moon;Kim, Seok-Hyun;Hwang, Jae-Bok;Yoon, Seong-Tak;Shim, Sang- In
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.2
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    • pp.134-139
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    • 2011
  • This study was conducted to evaluate the weed suppressing effects of two winter cover crops, hairy vetch and rye in foxtail millet and sorghum fields in 2010. Crop growth and development and weed occurrences in the fields were examined to know the efficiency of proposed cropping system. In hairy vetch treated plots, heading of minor cereals occurred early. The heading date was earlier by 1 day and 2 days in sorghum and foxtail millet, respectively. However, rye treatment delayed heading by 12 days and 8 days in sorghum and foxtail millet, respectively. Besides he effect of cover crop on ear emergence of crops, the Besides changed growth-related characteristics. Plant height, chlorophyll content and chlorophyll fluorescence of sorghum ere increased in hairy vetch treatment by 46.4% 88.7% and 7.9%, respectively. In foxtail millet, the characteristics ere also increased by 45.6%, 50.9% and 37.8%, respectively, s compared to control. Yields of sorghum and foxtail millet were increased by 105.1% and 135% as compared o control by hairy vetch treatment, respectively. However, he yields of cereal crops were decreased by rye cover crop treatment, the yields of sorghum and foxtail millet were decreased by 25.8% and 119.1%, respectively. Rye cover crop treatment inhibited crop growth suggesting nitrogen starvation in rye treated plots. In rye treatment, plant height, chlorophyll contents and chlorophyll fluorescence of sorghum ere slightly decreased by 7.1%, 10.8% and 10.8%, respectively, as compared to control whereas the inhibitory effects were greater in foxtail millet. Weed occurrences based n weed number in hairy vetch and rye plots were reduced n weed number in hairy vetch and rye plots were reduced y 27% and 20%, respectively. The smothering effect was weakened or disappeared after heading of crops. Weed number and dry weight in hairy vetch plot were increased by 159% and 55.2%, respectively, as compared to control. The results implied that weed suppressing of cover crops could be reduced drastically after heading of crops.

Assessment of cell adhesion, cell surface hydrophobicity, autoaggregation, and lipopolysaccharide-binding properties of live and heat-killed Lactobacillus acidophilus CBT LA1 (락토바실러스 아시도필러스 CBT LA1 생균과 사균체의 세포부착력, 자가응집력, 소수성 상호작용력, LPS 결합력에 대한 평가)

  • Shin, Joo-Hyun;Lee, Joong-Su;Seo, Jae-Gu
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.241-248
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    • 2015
  • Although studies on probiotics have been performed mostly with viable microbes, the beneficial functions of dead or heat-killed form of probiotic strains have also been examined. In this study, live and heat-killed forms of Lactobacillus acidophilus CBT LA1 were investigated in vitro and in vivo to evaluate the properties necessary for gut barrier protection. Cell surface hydrophobicity (CSH), autoaggregation (AA), cell adhesion, and lipopolysaccharide (LPS)-binding properties were evaluated. In addition, the suppressive effect on LPS-induced interleukin (IL)-8 expression was investigated in HT-29 cells. To identify optimal conditions for CBT LA1 to adhere to HT-29 cells, CBT LA1 cells were heat-treated at 80, 85, 90, 95, 100, or $121^{\circ}C$ for 10 min; cells treated at $80^{\circ}C$ for 10 min showed the highest adhesion. Heat-killed bacteria at $80^{\circ}C$ showed higher levels of LPS-binding, CSH, AA, adhesion to HT-29, and suppression of IL-8 expression than did live CBT LA1. In vivo imaging was performed to evaluate the ability of live or heat-killed CBT LA1 to remove LPS from the intestine in a rat model of infection. At 16 h after infection, fluorescence from FITC-conjugated LPS had mostly disappeared from the intestine of the rats administered with live or heat-killed CBT LA1; the effect was greater with heat-killed CBT LA1 at $80^{\circ}C$. These results suggest that heat-killed CBT LA1 as well as its live form can be applied as a pharmabiotic for protection of the gut barrier.