• Title/Summary/Keyword: SLC5A1

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Heterologous Expression of Human SLC1A5v2 as a Functional Glutamine Transporter in Escherichia coli

  • E Young Kim;Ji Won Park;Ok Bin Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.37-42
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    • 2023
  • Neutral and non-essential amino acid, glutamine (Gln), plays an essential role in supplying nitrogen to all the amino acids and nucleotides in the mammalian body. Gln is also the most important carbon source that provides intermediates for gluconeogenesis and fatty acid synthesis and supplements the tricarboxylic acid cycle in fast-growing cancer cells. Among the known 14 Gln transporter genes, soluted carrier family 1 member 5 (SLC1A5) has been reported to be closely associated with cancer cell growth. Three variants (v1, v2, and v3) have been derived from SLC1A5. Here, we established a heterologous gene expression system for the active form of human SLC1A5 variant-2 (hSLC1A5v2) in Escherichia coli. v2 is the smallest variant that has not yet been studied. Four expression systems were investigated: pBAD, pCold, pET, and pQE. We also addressed the problem of codon usage bias. Although pCold and pET overexpressed hSLC1A5v2 in E. coli, they were functionally inactive. hSLC1A5v2 using the pBAD system was able to catalyze the successful transport of Gln, even if it was not highly expressed. Initial activity of hSLC1A5v2 for [14C] Gln uptake in E. coli reached up to 6.73 μmole·min-1·gDW-1 when the cell was induced with 80 mM L-arabinose. In this study, we demonstrated a heterologous expression system for the human membrane protein, SLC1A5, in E. coli. Our results can be used for the functional comparison of SLC1A5 variants (v1, v2, and v3) in future studies, to facilitae the developement of SLC1A5 inhibitors as effective anticancer drugs.

Ginsenoside Rg5 promotes wound healing in diabetes by reducing the negative regulation of SLC7A11 on the efferocytosis of dendritic cells

  • Wei Xia;Zongdong Zhu;Song Xiang;Yi Yang
    • Journal of Ginseng Research
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    • v.47 no.6
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    • pp.784-794
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    • 2023
  • Background: ginsenoside Rg5 is a rare ginsenoside with known hypoglycemic effects in diabetic mice. This study aimed to explore the effects of ginsenoside Rg5 on skin wound-healing in the Leprdb/db mutant (db/db) mice (C57BL/KsJ background) model and the underlying mechanisms. Methods: Seven-week-old male C57BL/6J, SLC7A11-knockout (KO), the littermate wild-type (WT), and db/db mice were used for in vivo and ex vivo studies. Results: Ginsenoside Rg5 provided through oral gavage in db/db mice significantly alleviated the abundance of apoptotic cells in the wound areas and facilitated skin wound healing. 50 μM ginsenoside Rg5 treatment nearly doubled the efferocytotic capability of bone marrow-derived dendritic cells (BMDCs) from db/db mice. It also reduced NF-κB p65 and SLC7A11 expression in the wounded areas of db/db mice dose-dependently. Ginsenoside Rg5 physically interacted with SLC7A11 and suppressed the cystine uptake and glutamate secretion of BMDCs from db/db and SLC7A11-WT mice but not in BMDCs from SLC7A11-KO mice. In BMDCs and conventional type 1 dendritic cells (cDC1s), ginsenoside Rg5 reduced their glycose storage and enhanced anaerobic glycolysis. Glycogen phosphorylase inhibitor CP-91149 almost abolished the effect of ginsenoside Rg5 on promoting efferocytosis. Conclusion: ginsenoside Rg5 can suppress the expression of SLC7A11 and inhibit its activity via physical binding. These effects collectively alleviate the negative regulations of SLC7A11 on anaerobic glycolysis, which fuels the efferocytosis of dendritic cells. Therefore, ginsenoside Rg5 has a potential adjuvant therapeutic reagent to support patients with wound-healing problems, such as diabetic foot ulcers.

Expressional Analysis of Glucose Transporter Isoforms in the Efferent Ductules of Male Sprague Dawley Rat during Postnatal Development

  • Seo, Hee-Jung;Son, Chan-Wok;Lee, Ki-Ho
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.211-216
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    • 2009
  • A cell frequently utilizes glucose as a fuel of energy and a major substrate of lipid and protein syntheses. A regulation of glucose movement into and out of the cells is precisely controlled by cooperative works of passive and sodium-dependent active processes. At least 13 glucose cotransporter (Slc2a, GLUT) isoforms involve in passive movement of glucose in cells. The efferent ductules (EDs) play in a number of important functions for maintenance of male fertility. In the present study, using real-time PCR analysis, we determined gene expression of five Slc2a isoforms in the EDs. In addition, we compared expression levels of these Slc2a isoforms according to postnatal development ages, 1 week, 2 weeks, 1 month, and 3 months. Results from the current study showed that expression of Slc2a1, Slc2a3, and Slc2a5 mRNAs reached the highest levels at 1 month of age, followed by a transient decrease at 3 months of age. In addition, the level of Slc2a4 mRNA reminded at steady until 1 month of age and was significantly reduced at 3 months of age, whereas the highest level of Slc2a 8 mRNA was detected at 2 weeks of age. Data from the present study indicate a differential expression of various Slc2a isoforms in the ED according to postnatal ages. Thus, it is believed that glucose movement through the epithelial cells in the ED would be regulated by the coordinated manner among Slc2a isoforms expressed at a given age.

Maximization of The Number of Follicular Oocytes Recovered from The Bovine Ovaries (소 난소로부터 회수난포란수의 극대화 방법)

  • 유형진;최승철;이상호
    • Korean Journal of Animal Reproduction
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    • v.17 no.2
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    • pp.149-157
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    • 1993
  • A new technique was established to maximize the numbers of follicular oocytes recovered from the ovaries obtained at the slaughter house. And their further developmental capacity was demonstrated. There recovery techniques were used; aspiration (ASP, control), slicing (SLC) and slicing combining aspiration (ASP+SLC). Recovered oocytes were cultured in TCM 199+15% FCS+gonadotrophins in an atmosphere of 5% CO$_2$ in air at 39$^{\circ}C$ for 24 h. The nuclear maturation was detemined with chromo-some configuration by rapid staining. And cytoplasmic maturation was examined by the formation of female pronuclei with parthenogenetic activation of the matured oocyte after 18 h of co-culture with granulosa cell monolayer. Total 1,641 bovine follicular oocytes recovered from 245 ovaries. The number of oocytcs per ovary was 1.87 in ASP, 11.05 in SLC and 7.88 in ASP+SLC, respectively. SLC would yield 5.9 folds increase, compared with ASP. The rate of maturation were 92.9% in ASP, 79.1% in SLC and 71.7% in ASP+SLC, respectively. Although the maturation rate in ASP was the highest, metaphase II oocytes per ovary in SLC was 5 times higher than that of ASP. The rates of pronuclei formation upon ethanol activation were 75% in ASP, 67% in SLC and 62.5% in ASP+SLC, respectively. The results demonstrate that it should be possible to maximize the number of the follicular oocyte from the ovary for mass production of bovine embryos. Thus the established technique may provide efficient supply of bovine embryos for biochemical and molecular study of early bovine embryos.

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Ginsenoside Rd protects cerebral endothelial cells from oxygen-glucose deprivation/reoxygenation induced pyroptosis via inhibiting SLC5A1 mediated sodium influx

  • Li, Suping;Yu, Nengwei;Xu, Fei;Yu, Liang;Yu, Qian;Fu, Jing
    • Journal of Ginseng Research
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    • v.46 no.5
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    • pp.700-709
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    • 2022
  • Background: Ginsenoside Rd is a natural compound with promising neuroprotective effects. However, the underlying mechanisms are still not well-understood. In this study, we explored whether ginsenoside Rd exerts protective effects on cerebral endothelial cells after oxygen-glucose deprivation/reoxygenation (OGD/R) treatment and its potential docking proteins related to the underlying regulations. Method: Commercially available primary human brain microvessel endothelial cells (HBMECs) were used for in vitro OGD/R studies. Cell viability, pyroptosis-associated protein expression and tight junction protein degradation were evaluated. Molecular docking proteins were predicted. Subsequent surface plasmon resonance (SPR) technology was utilized for validation. Flow cytometry was performed to quantify caspase-1 positive and PI positive (caspase-1+/PI+) pyroptotic cells. Results: Ginsenoside Rd treatment attenuated OGD/R-induced damage of blood-brain barrier (BBB) integrity in vitro. It suppressed NLRP3 inflammasome activation (increased expression of NLRP3, cleaved caspase-1, IL-1β and GSDMD-N terminal (NT)) and subsequent cellular pyroptosis (caspase-1+/PI + cells). Ginsenoside Rd interacted with SLC5A1 with a high affinity and reduced OGD/R-induced sodium influx and potassium efflux in HBMECs. Inhibiting SLC5A1 using phlorizin suppressed OGD/R-activated NLRP3 inflammasome and pyroptosis in HBMECs. Conclusion: Ginsenoside Rd protects HBMECs from OGD/R-induced injury partially via binding to SLC5A1, reducing OGD/R-induced sodium influx and potassium efflux, thereby alleviating NLRP3 inflammasome activation and pyroptosis.

Restoration of Landsat ETM+ SLC-off Gaps Using SPOT Image (SPOT 영상을 이용한 Landsat-7의 SLC-off 영상 복원)

  • Kim Hye-Jin;Yu Ki-Yun;Kim Yong-Il
    • Proceedings of the Korean Society of Surveying, Geodesy, Photogrammetry, and Cartography Conference
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    • 2006.04a
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    • pp.229-234
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    • 2006
  • On May 31, 2003. Landsat 7 experienced an anomaly causing the Scan Line Corrector(SLC) to stop functioning normally. The SLC-off causes individual scan lines to alternately overlap and then leave large gaps at the edge of the Image. A many scientists with ongoing experience using ETM+ data evaluated the scientific usability and validity of Landsat 7 products containing the SLC anomaly The best reference scene for gap-filling is the other SLC-on Landsat scene that provide same resolution, few changes, and similar data acquisition. But receiving of Landsat imagery is not stable in Korea. So SPOT image can be another alternative solution because it is a steady-state multispectral satellite image as Landsat image. In this study, we filled the SLC-off gap s of 2, 3, 4 bands using SPOT image by a local regression technique, and assigned the optimum spectral value to gaps of 1, 5, 7 bands based on a spectral adjacency. Through this process, we could restore Landsat SLC-off image and evaluated the accuracy of the results.

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Genetic Polymorphisms of SLC8A1 Are Associated with Hypertension and Left Ventricular Hypertrophy in the Korean Population (한국인에서 SLC8A1의 유전적 다형성과 고혈압 및 좌심실 비대와 연관 연구)

  • Park, Hye-Jeong;Kim, Sung-Soo;Jin, Hyun-Seok
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.286-293
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    • 2019
  • Hypertension (HTN) is one of the major chronic diseases, and HTN is defined as being in a state of continuous high blood pressure. Left ventricular hypertrophy (LVH) is a condition in which the mass of the left ventricle has increased, and HTN is a leading cause of LVH. HTN and LVH are known to be caused by the interaction of environmental factors and genetic factors. It has been reported that the polymorphisms of SLC8A1, among the genetic factors that affect high blood pressure, are related to salt sensitivity hypertension. In this study, the genetic polymorphisms of SLC8A1 were chosen based on the Korean Genome and Epidemiology data. Logistic regression analysis was then performed for HTN and LVH. Linear regression analysis was also performed for systolic blood pressure (SBP) and diastolic blood pressure (DBP). As a result, 5 SNPs showed statistically significant associations (P<0.05) with HTN, and 10 SNPs showed statistically significant associations with LVH. rs1002671 and rs9789739 showed significant correlation at the same time with HTN and LVH. These results suggest that the polymorphisms of the SLC8A1 gene are linked to the development of HTN and LVH in Koreans. We expect these results to help us understand the pathogenic mechanisms for HTN and LVH.

Carrier frequency of SLC26A4 mutations causing inherited deafness in the Korean population

  • Kim, Hyogyeong;Lim, Hwan-Sub;Ryu, Jae-Song;Kim, Hyun-Chul;Lee, Sanghoo;Kim, Yun-Tae;Kim, Young-Jin;Lee, Kyoung-Ryul;Park, Hong-Joon;Han, Sung-Hee
    • Journal of Genetic Medicine
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    • v.11 no.2
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    • pp.63-68
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    • 2014
  • Purpose: The mutation of the SLC26A4 gene is the second most common cause of congenital hearing loss after GJB2 mutations. It has been identified as a major cause of autosomal recessive nonsyndromic hearing loss associated with enlarged vestibular aqueduct and Pendred syndrome. Although most studies of SLC26A4 mutations have dealt with hearing-impaired patients, there are a few reports on the frequency of these mutations in the general population. The purpose of this study was to evaluate the prevalence of SLC26A4 mutations that cause inherited deafness in the general Korean population. Materials and Methods: We obtained blood samples from 144 Korean individuals with normal hearing. The samples were subjected to polymerase chain reaction to amplify the entire coding region of the SLC26A4 gene, followed by direct DNA sequencing. Results: Sequencing analysis of this gene identified 5 different variants (c.147C>G, c.225G>C, c.1723A>G, c.2168A>G, and c.2283A>G). The pathogenic mutation c.2168A>G (p.H723R) was identified in 1.39% (2/144) of the subjects with normal hearing. Conclusion: These data provide information about carrier frequency for SLC26A4 mutation-associated hearing loss and have important implications for genetic diagnostic testing for inherited deafness in the Korean population.

Location-Aware Hybrid SLC/MLC Management for Compressed Phase-Change Memory Systems (압축 기반 상변화 메모리 시스템에서 저장 위치를 고려한 하이브리드 SLC/MLC 관리 기법)

  • Park, Jaehyun;Lee, Hyung Gyu
    • IEMEK Journal of Embedded Systems and Applications
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    • v.11 no.2
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    • pp.107-116
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    • 2016
  • Density of Phase-Change Memory (PCM) devices has been doubled through the employment of multi-level cell (MLC) technology. However, this doubled-capacity comes in the expense of severe performance degradation, as compared to the conventional single-level cell (SLC) PCM. This negative effect on the performance of the MLC PCM detracts from the potential benefits of the MLC PCM. This paper introduces an efficient way of minimizing the performance degradation while maximizing the capacity benefits of the MLC PCM. To this end, we propose a location-aware hybrid management of SLC and MLC in compressed PCM main memory systems. Our trace-driven simulations using real application workloads demonstrate that the proposed technique enhances the performance and energy consumption by 45.1% and 46.5%, respectively, on the average, over the conventional technique that only uses a MLC PCM.

Case report of cerebral creatine deficiency syndrome with novel mutation of SLC6A8 gene in a male child in Bangladesh

  • Rahman, Muhammad Mizanur;Fatema, Kanij
    • Journal of Genetic Medicine
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    • v.18 no.1
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    • pp.44-47
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    • 2021
  • Cerebral creatine deficiency syndrome (CCDS) is a disorder where a defect is present in transport of creatine in the brain. Creatine plays an essential role in the energy metabolism of the brain. This is a genetic disorder, autosomal recessive or X linked, characterized by intellectual disability, speech and language delay, epilepsy, hypotonia, etc. Until recently very few number of cases have been reported. Here we report a case of 1.5-year-old boy who had epilepsy (epileptic spasm and generalized tonic clonic seizure), intellectual disability, microcephaly, hypotonia and speech delay. His magnetic resonance imaging of brain showed cortical atrophy and electroencephalography showed burst suppression pattern. The diagnosis was confirmed by clinical exome sequencing which showed novel mutation of SLC6A8+ in exon 9, suggestive of X linked recessive CCDS. The patient was then treated with glycine, L-arginine and creatine monohydrate with multiple antiepileptic drugs.