• Research in Plant Disease
• /
• v.22 no.3
• /
• pp.178-183
• /
• 2016

Soybean yellow mottle mosaic virus (SYMMV) is a new emerging plant virus detected in soybean (Glycine max) in Korea. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid detection of SYMMV has been developed. In this study, we have designed primers (SYMM-F3/B3/FIP/BIP) specific to sequences from the coat protein gene of SYMMV genome. Sensitivity analysis showed that RT-LAMP was 10 to 100 times more sensitive than reverse transcription polymerase chain reaction (RT-PCR). The optimal reaction condition of RT-LAMP was determined at $65^{\circ}C$ for 50 minutes. The result indicates that RT-LAMP assay does not require special equipment and long time for SYMMV detection. Therefore, it can be an alternative detection method of RT-PCR in laboratory.

• Molecules and Cells
• /
• v.23 no.2
• /
• pp.161-169
• /
• 2007

We identified two alternatively spliced variants of the peroxisomal targeting signal 1 (PTS1) receptor protein Pex5ps in monocot (rice, wheat, and barley) but not in dicot (Arabidopsis and tobacco) plants. We characterized the molecular and functional differences between the rice (Oryza sativa) Pex5 splicing variants OsPex5pL and OsPex5pS. There is only a single-copy of OsPEX5 in the rice genome and RT-PCR analysis points to alternative splicing of the transcripts. Putative light-responsive cis-elements were identified in the 5' region flanking OsPEX5L and Northern blot analysis demonstrated that this region affected light-dependent expression of OsPEX5 transcription. Using the pex5-deficient yeast mutant Scpex5, we showed that OsPex5pL and OsPex5pS are able to restore translocation of a model PTS1 protein (GFP-SKL) into peroxisomes. OsPex5pL and OsPex5pS formed homo-complexes via specific interaction domains, and interacted with each other and OsPex14p to form hetero-complexes. Although overexpression of OsPex5pL in the Arabidopsis pex5 mutant (Atpex5) rescued the mutant phenotype, overexpression of OsPex5pS only resulted in partial recovery.

• Journal of Applied Biological Chemistry
• /
• v.58 no.3
• /
• pp.219-226
• /
• 2015

The anticancer activity of a methanolic extract from lemon leaves (MLL) was assessed in MCF-7-SC human breast cancer stem cells. MLL induced apoptosis in MCF-7-SC, as evidenced by increased apoptotic body formation, sub-G1 cell population, annexin V-positive cells, Bax/Bcl-2 ratio, as well as proteolytic activation of caspase-9 and caspase-3, and degradation of poly (ADP-ribose) polymerase (PARP) protein. Concomitantly, MLL induced the formation of acidic vesicular organelles, increased LC3-II accumulation, and reduced the activation of Akt, mTOR, and p70S6K, suggesting that MLL initiates an autophagic progression in MCF-7-SC via the Akt/mTOR pathway. Epithelial-mesenchymal transition (EMT), a critical step in the acquisition of the metastatic state, is an attractive target for therapeutic interventions directed against tumor metastasis. At low concentrations, MLL induced anti-metastatic effects on MCF-7-SC by inhibiting the EMT process. Exposure to MLL also led to an increase in the epithelial marker E-cadherin, but decreased protein levels of the mesenchymal markers Snail and Slug. Collectively, this study provides evidence that lemon leaves possess cytotoxicity and anti-metastatic properties. Therefore, MLL may prove to be beneficial as a medicinal plant for alternative novel anticancer drugs and nutraceutical products.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.29 no.1
• /
• pp.31-36
• /
• 2009

This study was conducted to find out an alternative way of rapid and accurate analysis of forage quality. Near reflectance infrared spectroscopy (NIRS) was used to evaluate the possibility of forage analysis and collect 258 samples such as barley for whole crop silage, forage corn and sudangrass from 2002 to 2007. The samples were analyzed for CP (crude protein), CF (crude fiber), ADF (acid detergent fiber), NDF (neutral detergent fiber) and IVTD (in vitro true digestibility), and also scanned using NIRSystem with wavelength from $400{\sim}2,400nm$. Multiple linear regression was used with wet analysis data for developing the calibration model and validate unknown samples. The important index In this experiment was SEC and SEP $r^2$ for CF, CP, NDF, ADF and IVTD in calibration set were 0.70, 0.86, 0.94, 0.94 and 0.89, also 0.47, 0.39, 0.89, 0.90 and 0.61 in validation sample, respectively. The results of this experiment indicates that NIRS was reliable analytical method to assess forage quality, specially in CF, ADF and IVTD, sample should be included for respective forage samples to get accurate result. More robust calibrations can be made to cover every forage samples if added representative sample set.

• Reproductive and Developmental Biology
• /
• v.30 no.1
• /
• pp.47-52
• /
• 2006

This study was conducted to examine whether the parthenogenetic mouse embryonic stem (P-mES) cells can differentiate into functional cardiomyocytes in vitro similar to (mES) cells. p-mES04 and IVF-derived mES03 cells were cultured by suspension culture for 4 days. The formed embryoid bodies (EBs) were treated with 0.75% dimethyl-sulfoxide (DMSO) for further 4 days (4-/4+), and then plated onto gelatin coated culture dish. The appearance of contracting cardiomyocytes from the P-mES04 and mES03 cells was examined for 30 days. The highest cumulative frequency was detected at days 13 (69.83%) and 22 (61.3%), respectively. By immunocytochemistry, beating P-mES04 cells were positively stained with muscle specific anti-sarcomeric a-actinin Ab and cardiac specific anti-cardiac troponin I Ab similar to contracted mES03 cells. When the expression of cardiac muscle-specific genes was analyzed by RT-PCR, beating P-mES04 cells were expressed cardiac specific L-type calcium channel, a1C, cardiac myosin heavy chain a, cardiac muscle heavy polypeptide $7{\beta}$, GATA binding protein 4 and atrial natriuretic factor, but not expressed skeletal muscle specific L-type calcium channel, a1S, which was similar to male adult heart cells and mES03-derived beating cardiomyocytes. The result demonstrates that the P-mES cells can be used as an alternative for the study on the characteristic analysis of in vitro cardiomyocyte differentiation from the ES cells.

• Korean Journal of Acupuncture
• /
• v.32 no.3
• /
• pp.108-115
• /
• 2015

Objectives : Acupuncture is frequently used as an alternative therapy for Parkinson's disease(PD) in Korea. Using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced Parkinson's disease mouse model, the present study investigated a possible role of acupuncture stimulation at GB34 in suppressing dopaminergic neuronal death and regulating the phosphorylation of protein kinase B(Akt) in substantia nigra(SN) and striatum(ST). Methods : Eight-week-old male C57BL/6 mice were administered intraperitoneally with 30 mg/kg of MPTP at 24-h intervals for 5 days. Acupuncture stimulation at GB34 or SI3 was performed once a day for 12 days consecutively from the first MPTP injection. After the last acupuncture stimulation, pole test was performed to assess the effect of the acupuncture stimulations. Dopaminergic neuronal survival in the SN and the ST, dopamine transporter( DAT) and caspase-3 expression in the ST were evaluated by immunohistochemistry. The phosphorylations of Akt in the SN and the ST were measured by Western blotting. Results : MPTP administration caused behavioral impairment and dopaminergic neuronal death in the SN and the ST. It also decreased DAT expression and increased caspase-3 expression in the ST. Acupuncture stimulation at GB34 alleviated these MPTP-induced impairments. Moreover, MPTP suppressed Akt phosphorylation in the SN and the ST, whereas acupuncture stimulation at GB34 alleviated the phosphorylation in the SN. Conclusions : These results indicate that acupuncture stimulation at GB34 can inhibit MPTP-induced dopaminergic neuronal death and alleviate the Akt phosphorylation in the SN, suggesting a possible role for acupuncture in the treatment of PD.

• Korean Journal of Optics and Photonics
• /
• v.20 no.6
• /
• pp.346-353
• /
• 2009

Glucose and protein in urine are among the important substances for urine analysis and have generally been measured based on a reagent strip test. In this study, these two substances were measured using mid-infrared absorption spectroscopy. Samples were prepared from a commercial synthetic urine product. Glucose and albumin were added as well as red blood cells, which are expected to create the most spectroscopic interference of any substance. Concentrations of these substances were varied independently. Optimal wavelength regions were determined from a partial least squares regression analysis (glucose 980 - 1150/cm, albumin 1400 - 1570/cm). Interference by other substances increased the differences between measured and predicted values. Albumin measurement in particular weres heavily influenced by the presence of glucose and red blood cells. Depending on the inference by other substances, measurement errors were 29.85${\sim}$45.19 mg/dl for a glucose level between 0 and 1000 mg/dl and 14.0${\sim}$93.11 mg/dl for an albumin level of 0 ${\sim}$ 500 mg/dl. Our study proposes an alternative to the chemical test-strip analysis, which shows only discrete concentration levels.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.31 no.4
• /
• pp.409-414
• /
• 2011

This study was conducted to find out an alternative way of rapid and accurate analysis of forage quality. Near Infrared Reflectance Spectroscopy (NIRS) was used to evaluate the possibility of forage analysis. 175 samples consisted of Italian ryegrass, whole crop barley and pea seeded spring in 2009 were collected. The samples were analyzed for moisture, crude protein (CP), crude ash (CA), acid detergent fiber (ADF), and neutral detergent fiber (NDF), and also scanned using NIRSystem with wavelength from 400~2,500 nm. Multiple linear regression was used with wet analysis data for developing the calibration model and validated unknown samples. The important index in this experiment were SEC, SEP. The r2 value for moisture, CP, CA, ADF, and NDF in calibration set was 0.65, 0.97, 0.93, 0.99, and 0.97 and also was 0.15, 0.94, 0.96, 0.98 and 0.98 in validation set, respectively. The results of this experiment indicates that NIRS was reliable analytical method to assess forage quality for CP, CA ADF and NDF except moisture content in forage when proper samples incorporated into the equation development.

• Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
• /
• v.32 no.1
• /
• pp.15-23
• /
• 2021

Background and Objectives During speech, the vocal folds oscillate at frequencies ranging from 100-200 Hz with amplitudes of a few millimeters. Mechanical stimulation is an essential factor which affects metabolism of human vocal folds. The effect of mechanical vibration on the cellular response in the human vocal fold fibroblasts cells (hVFFs) was evaluated. Materials and Method We created a culture systemic device capable of generating vibratory stimulations at human phonation frequencies. To establish optimal cell culture condition, cellular proliferation and viability assay was examined. Quantitative real time polymerase chain reaction was used to assess extracellular matrix (ECM) related and growth factors expression on response to changes in vibratory frequency and amplitude. Western blot was used to investigate ECM and inflammation-related transcription factor activation and its related cellular signaling transduction pathway. Results The cell viability was stable with vibratory stimulation within 24 h. A statistically significant increase of ECM genes (collagen type I alpha 1 and collagen type I alpha 2) and growth factor [transforming growth factor β1 (TGF-β1) and fibroblast growth factor 1 (FGF-1)] observe under the experimental conditions. Vibratory stimulation induced transcriptional activation of NF-κB by phosphorylation of p65 subunit through cellular Mitogen-activated protein kinases activation by extracellular signal regulated kinase and p38 mitogen-activated protein kinases (MAPKs) phosphorylation on hVFFs. Conclusion This study confirmed enhancing synthesis of collagen, TGF-β1 and FGF was testified by vibratory stimulation on hVFFs. This mechanism is thought to be due to the activation of NF-κB and MAPKs. Taken together, these results demonstrate that vibratory bioreactor may be a suitable alternative to hVFFs for studying vocal folds cellular response to vibratory vocalization.

• Korean Chemical Engineering Research
• /
• v.60 no.3
• /
• pp.398-406
• /
• 2022

The outer membrane of Gram-negative bacteria is the outermost layer of cellular environment in which numerous biophysical and biochemical processes are in action sustaining viability. Advances in cell engineering enable modification of bacterial genetic information that subsequently alters membrane physiology to adapt bacteria to specific purposes. Surface display of a functional molecule on the outer membranes is one of strategies that directs host cells to respond to a specific extracellular matter or stimulus. While intracellular expression of a functional peptide or protein fused to a membrane-anchoring motif is commonly practiced for surface display, the method is not readily applicable to exogenous or large proteins inexpressible in bacteria. Chemical conjugation at reactive groups naturally occurring on the membrane might be an alternative, but often compromises fitness due to non-specific modification of essential components. Herein, we demonstrated two distinct approaches that enable site-specific decoration of the outer membrane with a fluorescent agent in Escherichia coli. An unnatural amino acid genetically incorporated in a surface-exposed peptide could act as a chemoselective handle for bioorthogonal dye labeling. A surface-displayed α-helical domain originating from a part of a selected heterodimeric coiled-coil complex could recruit and anchor a green fluorescent protein tagged with a complementary α-helical domain to the membrane surface in a site- and hetero-specific manner. These methods hold a promise as on-demand tools to confer new functionalities on the bacterial membranes.