Hak Yeong Kim, Kain Seo, Hong Jin Jeon, Unjoo Lee, and Hyosang LeeMol. Cells 2017; 40(8): 523-532 https://doi.org/10.14348/molcells.2017.0153
Abstract : Functional near-infrared spectroscopy (fNIRS) is a noninvasive optical imaging technique that indirectly assesses neuronal activity by measuring changes in oxygenated and deoxygenated hemoglobin in tissues using near-infrared light. fNIRS has been used not only to investigate cortical activity in healthy human subjects and animals but also to reveal abnormalities in brain function in patients suffering from neurological and psychiatric disorders and in animals that exhibit disease conditions. Because of its safety, quietness, resistance to motion artifacts, and portability, fNIRS has become a tool to complement conventional imaging techniques in measuring hemodynamic responses while a subject performs diverse cognitive and behavioral tasks in test settings that are more ecologically relevant and involve social interaction. In this review, we introduce the basic principles of fNIRS and discuss the application of this technique in human and animal studies.
Moon-Soo Kim, and Anu Ganesh KiniMol. Cells 2017; 40(8): 533-541 https://doi.org/10.14348/molcells.2017.0139
Abstract : Engineered DNA-binding domains provide a powerful technology for numerous biomedical studies due to their ability to recognize specific DNA sequences. Zinc fingers (ZF) are one of the most common DNA-binding domains and have been extensively studied for a variety of applications, such as gene regulation, genome engineering and diagnostics. Another novel DNA-binding domain known as a transcriptional activator-like effector (TALE) has been more recently discovered, which has a previously undescribed DNA-binding mode. Due to their modular architecture and flexibility, TALEs have been rapidly developed into artificial gene targeting reagents. Here, we describe the methods used to design these DNA-binding proteins and their key applications in biomedical research.
Bo Long, Tian-Yi Gan, Rong-Cheng Zhang, and Yu-Hui ZhangMol. Cells 2017; 40(8): 542-549 https://doi.org/10.14348/molcells.2017.0012
Abstract : Cardiomyocyte apoptosis is initiated by various cellular insults and accumulated cardiomyocyte apoptosis leads to the pathogenesis of heart failure. Excessive reactive oxygen species (ROS) provoke apoptotic cascades. Manganese superoxide dismutase (MnSOD) is an important antioxidant enzyme that converts cellular ROS into harmless products. In this study, we demonstrate that MnSOD is down-regulated upon hydrogen peroxide treatment or ischemia/reperfusion (I/R) injury. Enhanced expression of MnSOD attenuates cardiomyocyte apoptosis and myocardial infarction induced by I/R injury. Further, we show that miR-23a directly regulates the expression of MnSOD. miR-23a regulates cardiomyocyte apoptosis by suppressing the expression of MnSOD. Our study reveals a novel model regulating cardiomyocyte apoptosis which is composed of miR-23a and MnSOD. Our study provides a new method to tackling apoptosis related cardiac diseases.
Sun-Yi Hyun, Ji-Hye Lee, Kyung-Jung Kang, and Young-Joo JangMol. Cells 2017; 40(8): 550-557 https://doi.org/10.14348/molcells.2017.0019
Abstract : The periodontal ligament (PDL) is the connective tissue between tooth root and alveolar bone containing mesenchymal stem cells (MSC). It has been suggested that human periodontal ligament stem cells (hPDLSCs) differentiate into osteo/cementoblast and ligament progenitor cells. The periodontitis is a representative oral disease where the PDL tissue is collapsed, and regeneration of this tissue is important in periodontitis therapy. Fibroblast growth factor-2 (FGF-2) stimulates proliferation and differentiation of fibroblastic MSCs into various cell lineages. We evaluated the dose efficacy of FGF-2 for cytodifferentiation of hPDLSCs into ligament progenitor. The fibrous morphology was highly stimulated even at low FGF-2 concentrations, and the expression of teno/ligamentogenic markers, scleraxis and tenomodulin in hPDLSCs increased in a dose dependent manner of FGF-2. In contrast, expression of the osteo/cementogenic markers decreased, suggesting that FGF-2 might induce and maintain the ligamentogenic potential of hPDLSCs. Although the stimulation of tenocytic maturation by TGF-β1 was diminished by FGF-2, the inhibition of the expression of early ligamentogenic marker by TGF-β1 was redeemed by FGF-2 treatment. The stimulating effect of BMPs on osteo/cementogenesis was apparently suppressed by FGF-2. These results indicate that FGF-2 predominantly differentiates the hPDLSCs into teno/ligamentogenesis, and has an antagonistic effect on the hard tissue differentiation induced by BMP-2 and BMP-4.
Myungook Lee, Jong Il Ahn, Ah Ran Lee, Dong Woo Ko, Woo Sub Yang, Gene Lee, Ji Yeon Ahn, and Jeong Mook LimMol. Cells 2017; 40(8): 558-566 https://doi.org/10.14348/molcells.2017.0058
Abstract : Regular monitoring on experimental animal management found the fluctuation of ART outcome, which showed a necessity to explore whether superovulation treatment is responsible for such unexpected outcome. This study was subsequently conducted to examine whether superovulation treatment can preserve ultrastructural integrity and developmental competence of oocytes following oocyte activation and embryo culture. A randomized study using mouse model was designed and in vitro development (experiment 1), ultrastructural morphology (experiment 2) and functional integrity of the oocytes (experiment 3) retrieved after PMSG/hCG injection (superovulation group) or not (natural ovulation; control group) were evaluated. In experiment 1, more oocytes were retrieved following superovulation than following natural ovulation, but natural ovulation yielded higher (p < 0.0563) maturation rate than superovulation. The capacity of mature oocytes to form pronucleus and to develop into blastocysts in vitro was similar. In experiment 2, a notable (p < 0.0186) increase in mitochondrial deformity, characterized by the formation of vacuolated mitochondria, was detected in the superovulation group. Multivesicular body formation was also increased, whereas early endosome formation was significantly decreased. No obvious changes in other microorganelles, however, were detected, which included the formation and distribution of mitochondria, cortical granules, microvilli, and smooth and rough endoplasmic reticulum. In experiment 3, significant decreases in mitochondrial activity, ATP production and dextran uptake were detected in the superovulation group. In conclusion, superovulation treatment may change both maturational status and functional and ultrastuctural integrity of oocytes. Superovulation effect on preimplantation development can be discussed.
Yoon-Jin Lee, Jin-Ho Bae, Soo-A Kim, Sung-Ho Kim, Kee-Min Woo, Hae-Seon Nam, Moon-Kyun Cho, and Sang-Han LeeMol. Cells 2017; 40(8): 567-576 https://doi.org/10.14348/molcells.2017.0059
Abstract : The Na+/H+ exchanger is responsible for maintaining the acidic tumor microenvironment through its promotion of the reabsorption of extracellular Na+ and the extrusion of intracellular H+. The resultant increase in the extracellular acidity contributes to the chemoresistance of malignant tumors. In this study, the chemosensitizing effects of cariporide, a potent Na+/H+-exchange inhibitor, were evaluated in human malignant mesothelioma H-2452 cells preadapted with lactic acid. A higher basal level of phosphorylated (p)-AKT protein was found in the acid-tolerable H-2452AcT cells compared with their parental acid-sensitive H-2452 cells. When introduced in H-2452AcT cells with a concentration that shows only a slight toxicity in H-2452 cells, cariporide exhibited growth-suppressive and apoptosis-promoting activities, as demonstrated by an increase in the cells with pyknotic and fragmented nuclei, annexin V-PE(+) staining, a sub-G0/G1 peak, and a G2/M phase-transition delay in the cell cycle. Preceding these changes, a cariporide-induced p-AKT down-regulation, a p53 up-regulation, an ROS accumulation, and the depolarization of the mitochondrial-membrane potential were observed. A pretreatment with the phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002 markedly augmented the DNA damage caused by the cariporide, as indicated by a much greater extent of comet tails and a tail moment with increased levels of the p-histone H2A.X, p-ATMSer1981, p-ATRSer428, p-CHK1Ser345, and p-CHK2Thr68, as well as a series of pro-apoptotic events. The data suggest that an inhibition of the PI3K/AKT signaling is necessary to enhance the cytotoxicity toward the acid-tolerable H-2452AcT cells, and it underlines the significance of proton-pump targeting as a potential therapeutic strategy to overcome the acidic-microenvironment-associated chemotherapeutic resistance.
Chieun Song, Taeyoon Kim, Woo Sik Chung, and Chae Oh LimMol. Cells 2017; 40(8): 577-586 https://doi.org/10.14348/molcells.2017.0075
Abstract : Phytocystatins (PhyCYSs) are plant-specific proteinaceous inhibitors that are implicated in protein turnover and stress responses. Here, we characterized a PhyCYS from
Taewook Kim, June Hyun Park, Sang-gil Lee, Soyoung Kim, Jihyun Kim, Jungho Lee, and Chanseok ShinMol. Cells 2017; 40(8): 587-597 https://doi.org/10.14348/molcells.2017.0086
Abstract : MicroRNAs (miRNAs) are essential small RNA molecules that regulate the expression of target mRNAs in plants and animals. Here, we aimed to identify miRNAs and their putative targets in
Subash Marasini, Da-Young Chang, Jin-Hwa Jung, Su-Jung Lee, Hye Lim Cha, Haeyoung Suh-Kim, and Sung-Soo KimMol. Cells 2017; 40(8): 598-605 https://doi.org/10.14348/molcells.2017.0095
Abstract : Human mesenchymal stem cells (MSCs) are currently being evaluated as a cell-based therapy for tissue injury and degenerative diseases. Recently, several methods have been suggested to further enhance the therapeutic functions of MSCs, including genetic modifications with tissue- and/or disease-specific genes.The objective of this study was to examine the efficiency and stability of transduction using an adenoviral vector in human MSCs. Additionally, we aimed to assess the effects of transduction on the proliferation and multipotency of MSCs. The results indicate that MSCs can be transduced by adenoviruses
(A) Fluorescence-activated cell-sorting analysis of MSCs transduced with GFP-expressing adenovirus (1–200 multiplicity of infection [MOI]) and untransduced controls. (B) Microscopy images of GFP expression in MSCs transduced at 200 MOI. Scale bar = 50 μm. (C) The percentage of GFP positive cells with increasing MOI.|@|~(^,^)~|@|Transgene expression in Ad-GFP MSCs.
(A) Representative fluorescence images of transduced cells at 2 and 12 days post-transduction. GFP-positive cells display a larger cell size with flattened morphology (arrows). Scale bars = 50 μm (B-D) The total number of cells, frequency of GFP-positive cells, and mean GFP intensity were measured using flow cytometry. (E) Senescence-associated β-galactosidase activity was examined in transduced MSCs (MOI 50) and cultured for one passage. Arrows indicated positive X-gal staining. Scale bars = 50 μm.|@|~(^,^)~|@|Surface antigen expression profiles of Ad-GFP MSCs.
MSCs transduced with GFP-expressing adenovirus (MOI at 50) were examined for MSC surface marker expression at 2 days post-transduction. Isotype antibodies served as negative control.|@|~(^,^)~|@|Mesodermal differentiation potential of Ad-GFP MSCs.
(A, B) Osteogenic differentiation was assessed by alizarin red S staining to visualize calcium deposits. (C, D) Chondrogenic differentiation was assessed by Alcian blue and immunohistochemical staining to detect glycosaminoglycans and GFP expression, respectively. Dark brown colored cells in (D) indicate GFP-positive cells. (E, F) Adipogenesis was monitored for intracellular lipid accumulation by microscopy and (G, H) oil-red O staining. All photomicrographs are the merges of GFP and bright field images. Scale bars = 50 μm.|@|~(^,^)~|@|Transgene stability in Ad-GFP MSCs under non-dividing culture conditions.
Long-term transgene stability was monitored after 30 days of culturing under conditions unfavorable to cell division (i.e, high-density plating with limited passaging). GFP expression was then assessed by fluorescence microscopy. Arrows indicate representative cells.