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Biology subjects

Yoon, J.-H.

Publications and source records attributed to Yoon, J.-H..

9 recordsLinked to original sources

Neat1 alleviates ischemic injury in skeletal muscles

Peripheral artery disease (PAD) has historically been regarded as an age-related vascular disorder; however, recent attention has shifted toward the myopathic components of the disease. Conventional interventions, such as revascularization, have had limited success in reversing muscle pathology or preventing adverse outcomes like amputation. Hypoxia-inducible factors (HIFs) are key regulators of cellular responses to ischemia, including the promotion of angiogenesis and glycolysis. While pharmacological stabilization of HIF proteins represents a promising therapeutic strategy, its efficacy is diminished in aged muscles due to their intrinsically low basal HIF expression. We hypothesized that long noncoding RNAs (lncRNAs) might enhance the hypoxic response in aged muscle through post-transcriptional regulation of HIF expression. Our study identified a long noncoding RNA Neat1, as a critical mediator of the hypoxia-induced stress response, including upregulation of Hif1. In a murine hindlimb ischemia model, Neat1 knockout mice exhibited extensive necrosis following femoral artery ligation, whereas Neat1 overexpression conferred protection against ischemic injury. Mechanistically, we found that Neat1 regulates the stability of Hif1a mRNA, as Hif1a transcript levels were significantly reduced in Neat1-deficient muscle cells. Importantly, aged muscles displayed a blunted hypoxic response due to diminished Hif1a expression--an effect that was reversed through Neat1 overexpression, resulting in improved resistance to ischemic damage. In summary, our findings highlight Neat1 as a novel regulator of muscle adaptation to hypoxia in aging. Enhancing Neat1 expression may represent a promising therapeutic strategy for improving ischemic outcomes in patients with peripheral artery disease.

cell biology↗

Spatiotemporal regulation by downstream genes of Prok2 in olfactory system: from development to function

Olfaction is important for the quality of life; however, in Kallmann syndrome (KS), defective development results in olfactory dysfunction. The mechanism underlying olfactory development, especially in the olfactory epithelium (OE), which detects olfactory signals, remains unclear. Mutations in PROK2, which encodes prokineticin-2, occur in approximately 9% of the KS patients with olfactory defects. In this study, we examined olfactory function and analyzed the causes of olfactory dysfunction based on spatiotemporal development and gene expression changes in Prok2 knockout (KO) model mice with KS. Adult Prok2 KO mice exhibited reduced ability to detect olfactory signals. Maturation of olfactory sensory neurons (OSNs) in the OE and formation of glomeruli in the olfactory bulb (OB) in adult Prok2 KO mice were disrupted, thus causing olfactory dysfunction. Molecular analysis of Prok2 KO mice during embryonic development revealed abnormal development of OB layers and reduced differentiation to mature OSNs in the OE, which caused defects in the entire olfactory system. Downstream signaling genes of Prok2, including intermediate filament genes and genes expressed in the putative OB, mediated olfactory system organization. Our findings reveal the role of Prok2 in olfactory system organization, providing insights into the mechanisms through which olfactory development defects translate to olfactory function.

developmental biology↗

Mature microRNA-binding protein QKI promotes microRNA-mediated gene silencing

Although Argonaute (AGO) proteins have been the focus of microRNA (miRNA) studies, we observed AGO-free mature miRNAs directly interacting with RNA-binding proteins, implying the sophisticated nature of fine-tuning gene regulation by miRNAs. To investigate microRNA-binding proteins (miRBPs) globally, we analyzed PAR-CLIP data sets to identify RBP quaking (QKI) as a novel miRBP for let-7b. Potential existence of AGO-free miRNAs were further verified in genetically engineered AGO-depleted human and mouse cells. We have shown that QKI serves as an auxiliary factor empowering AGO2/let-7b-mediated gene silencing. Depletion of QKI decreases interaction of AGO2 with let-7b and target mRNA, consequently controlling target mRNA decay. QKI, however, also suppresses the dissociation of let-7b from AGO2, and slows assembly of AGO2/miRNA/target mRNA complexes at the single-molecule level. We also revealed that QKI suppresses cMYC expression at post-transcriptional level, and decreases proliferation and migration of HeLa cells, demonstrating that QKI is a tumor suppressor gene by in part augmenting let-7b activity. Our data show that QKI is a new type of RBP implicated in the versatile regulation of miRNA-mediated gene silencing.

biochemistry↗

Characteristics of viable but nonculturable Vibrio parahaemolyticus induced under extended periods of cold-starvation with various NaCl concentrations

This study was undertaken to examine the induction of VBNC states of Vibrio parahaemolyticus under prolonged cold-starvation with various NaCl concentrations and their responsive characteristics to maintain cell viability. V. parahaemolyticus entered the viable but nonculturable (VBNC) state in artificial sea water at 4{degrees}C within 80 day and persisted in the VBNC state for 150 days. During cold-starvation, bacterial cells were used to estimate their cell functions, including cytotoxicity, fatty acid composition, membrane potential, and morphology. Cytotoxic effect of V. parahaemolyticus cells against animal cell lines was decreased to below 50% after 80 days. VBNC V. parahaemolyticus cells showed decreasing levels of palmitic, vaccenic, and hexadecenoic acid on membrane, concomitantly with the formation of empty gaps between the cytoplasmic and outer membrane, in comparison with those of the pure cultures. Starvation at 4{degrees}C for 30 days resulted in a high increase in N-phenyl-1-napthylamine intensity within V. parahaemolyticus cells. Membrane potential and cellular composition were strongly affected by increasing NaCl contents of the microcosms after its evolution into the VBNC state. VBNC V. parahaemolyticus cells may undergo selected physiological changes such as the modulation of membrane potential and re-arrangement of cellular composition.

microbiology↗

Increased capillary stalling is associated with endothelial glycocalyx loss in subcortical vascular dementia

Proper regulation and patency of cerebral microcirculation is crucial for maintaining a healthy brain. Capillary stalling, i.e., the brief interruption of microcirculation mainly by leukocytes, has been observed in several diseases and contributes to disease pathogenesis or progression. However, the underpinning mechanism for leukocyte capillary plugging remains elusive. Therefore, we investigated the mechanism of capillary stalling in mice during the development of subcortical vascular dementia (SVaD), the most common type of vascular dementia characterized by impaired microcirculation and associated pathological features. Longitudinal optical coherence tomography angiography showed increased number of stalled segments as the disease progressed, while two-photon microscopy indicated a less extensive endothelial glycocalyx (EG) in the stalled segments. We also found that increased gliosis and blood-brain barrier leakage were correlated with the increased number of stalled segments. Based on the above, we conclude that EG potentially mediates capillary stalling and can be a therapeutic target of SVaD.

neuroscience↗

Differential TM4SF5-mediated SIRT1 modulation and signaling for chronic liver disease

Here we show the roles of transmembrane 4 L six family member 5 (TM4SF5) in the progression of nonalcoholic steatosis (or NAFL) to steatohepatitis (NASH). The overexpression of TM4SF5 caused nonalcoholic steatosis and NASH in an age-dependent manner. Initially, TM4SF5-positive hepatocytes and livers exhibited lipid accumulation, decreased SIRT1, increased SREBPs levels, and inactive STAT3 via SOCS1/3 upregulation. In older animals, TM4SF5 under an inflammatory environment increased SIRT1 expression and STAT3 activity with no significant change to SOCSs and SREBPs levels, leading to active STAT3-mediated fibrotic extracellular matrix (ECM) production. Liver tissues from clinical human patients with NAFL or NASH also showed such a TM4SF5-SIRT1-STAT3-ECM relationship correlated with fibrosis score and age. Ligand-independent and TM4SF5-mediated STAT3 activity led to collagen I and laminins/laminin {gamma}2 expression in hepatic stellate cells and hepatocytes, respectively. Laminin {gamma}2 suppression abolished CCl4-mediated liver damage and ECM production and reduced SIRT1 and active-STAT3, but did not alter SREBP1 or SOCSs levels. These findings suggest that TM4SF5, CCL20, SIRT1, and/or laminin {gamma}2 may be promising therapeutic targets against liver disease.

cell biology↗

SMAD3 Determines Conventional versus Plasmacytoid Dendritic Cell Fates

Transforming growth factor (TGF)-{beta} plays crucial roles in differentiation of dendritic cells (DC). However, molecular mechanisms how TGF-{beta} regulates DC differentiation remain largely unknown. Here, we show that selective repression of one of the TGF-{beta} receptor-regulated SMADs (R-SMADs), SMAD3 directs conventional DC (cDC) differentiation, whereas maintenance of SMAD3 is indispensable for plasmacytoid DC (pDC) differentiation. Expression of SMAD3 was specifically downregulated in CD115+ common DC progenitor (CDP), pre-cDCs and cDCs. SMAD3 deficient mice showed a significant reduction in pre-pDCs and pDCs with increased CDP, pre-cDCs and cDCs. SMAD3 upregulated the pDC-related genes: SPI-B, E2-2 and IKAROS, while it repressed FLT3 and the cDC-related genes: IRF4 and ID2. STAT3 and a SMAD transcriptional co-repressor, c-SKI repressed SMAD3 for cDC differentiation, whereas canonical SMAD-mediated TGF-{beta} signalling maintained SMAD3 for pDC differentiation. Thus, SMAD3 is the pivotal determinant to bifurcate cDC and pDC differentiation in the steady-state condition.

immunology↗

Physiological Characteristics of Viable-but-nonculturable Vibrio parahaemolyticus upon Prolonged Exposure to the Refrigerator Temperature

Although it has been reported that viable-but-nonculturable (VBNC) cells of Vibrio parahaemolyticus can be developed by a prolonged duration of cold-starvation there are restricted cellular characteristics available on understanding the exact mechanisms governing the entry of pathogens into the VBNC state. Therefore, this research was aimed at determining the cellular profile of VBNC cells of V. parahaemolyticus upon exposure to the refrigerator temperature. Strains of V. parahaemolyticus were incubated in artificial sea water (ASW) microcosms (pH 6) added with different amounts of NaCl at 4{degrees}C until these pathogens entered into such a dormant state. At a regular time-interval, both culturability and viability of these bacteria were enumerated, and then cellular profiling were carried out in terms of cellular membrane permeability, enzymatic activity, hydrophobicity, fatty acid composition, and morphological changes after cells of V. parahaemolyticus became the VBNC state. Three strains of V. parahaemolyticus used in this study showed that VBNC cells retained the strong virulent properties to Vero and CACO-2 cell lines, re-gained the cytotoxicity even after resuscitation, became permeabilized in terms of the outer membrane, showed lower levels of enzymatic (catalase and glutathione-S-transferase) activities, exerted the increasing hydrophobicity, and then exhibited increasing amounts of saturated fatty acids.\n\nIMPORTANCETo the current best knowledge, there are restricted information available on understanding the physiological characterization of viable-but-nonculturable cells. Most previous studies are still making a degree of efforts in discovering the causative effector causing microorganisms to be induced into the VBNC state. Herein, the present study showed that pathogenic V. parahaemolyticus can enter into the VBNC state when challenged by a certain environmental stress where higher amounts of NaCl combined with acidic pHs was artificially controlled. Importantly, it was indicated that VBNC V. parahaemolyticus maintained peculiarly different physiological characteristics. Furthermore, this study proposed a novel approach on the transient/stepwise conversion of the bacteria into the VBNC state. Specific alternative tools for measuring and controlling the incidence of VBNC pathogens on food are not established until now. In this aspect, results obtained from this study will used to provide an effective insight in determining physiological properties of viable-but-nonculturable V. parahaemolyticus.

microbiology↗

Analysis of the Resuscitation-Availability of Viable-But-Nonculturable Cells of Vibrio parahaemolyticus upon Exposure to the Refrigerator Temperature

Major pathogenic strains of Vibrio parahaemolyticus can enter into the viable-but-nonculturable (VBNC) state when subjected to environmental conditions commonly encountered during food processing. Especially, VBNC cells can be recovered to the culturable state reversibly by removing the causative stress, expressing higher levels of virulence factors. Therefore, the aim of this study was to determine if VBNC V. parahaemolyticus strains retain the resuscitation-availability upon eliminating the adverse condition, followed by the enrichment in developed resuscitation-facilitating buffers. Bacterial cells were shown to enter into the VBNC state in artificial sea water (ASW, pH 6) microcosms at 4{degrees}C within 70 days. VBNC cells were harvested, inoculated in formulated resuscitation-buffers, and then incubated at 25{degrees}C for several days. TSB (pH 8) supplemented with 3% NaCl (TSBA) exhibited the higher resuscitation-availability of VBNC cells. It was also shown that TSBA containing 10,000 U/mg/protein catalase, 2% sodium pyruvate, 20 mM MgSO4, 5 mM ethylenediaminetetraacetic acid (EDTA), and cell free supernatants extracted from the pure cultures of V. parahaemolyticus was more effective in resuscitating VBNC cells of V. parahaemolyticus, showing by 7.69-8.91 log10 CFU/ml.\n\nIMPORTANCEGenerally, higher concentrations ([≤]40%) of NaCl are used for preserving different sorts of food products from bacterial contaminations. However, it was shown from the present study that strains of V. parahaemolyticus were able to persist in maintaining the cellular viability, thereby entering into the VBNC state upon exposure to the refrigerator temperature for 80 days. Hence, the ability of VBNC V. parahaemolyticus to re-enter into the culturable state was examined, using various resuscitation buffers that were formulated in this study. VBNC cells re-gained the culturability successfully when transferred onto the resuscitation-buffer D, and then incubated at 25{degrees}C for several days. Resuscitation-facilitating agent D is consisting of antioxidizing agents, mineral, an emulsifier, and cell free supernatants from the actively growing cells of V. parahaemolyticus. It appeared that such a reversible conversion of VBNC cells to the culturable state would depend on multiple resuscitation-related channels.

microbiology↗