• 대한의생명과학회지
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• 제15권1호
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• pp.55-60
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• 2009

Insect cells such as Spodoptera frugiperda Clone 9 (Sf9) cells are widely chosen as the host for heterologous expression of a mammalian sugar transport protein using the baculovirus expression system. Characterization of the expressed protein is expected to include assay of its function, including its ability to transport sugars and to bind inhibitory ligands such as cytochalasin B. It is therefore very important first to establish the transport characteristics and other properties of the endogenous sugar transport proteins of the host insect cells. However, very little is known of the transport characteristics of Sf9 cells, although their ability to grow on TC-100 medium strongly suggested the presence of endogenous glucose transport system. In order to investigate the substrate and inhibitor recognition properties of the Sf9 cell transporter, the ability of pentoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. To determine the time period over which of sugar into the Sf cells was linear, the uptake of 2dGlc 0.1mM extracellular concentration was measured over periods ranging from 30 seconds to 30 minutes. The uptake was linear for at least 2 minutes at the concentration, implying that uptake made over a 1 minute time course would reflect initial rates of the sugar uptake. The data have also revealed the existence of a saturable transport system for pentose uptake by the insect cells. The transport was inhibited by D-xylose and D-ribose, although not as effective as hexoses. However, L-xylose had a little effect on 2dGlc transport in the Sf9 cells, indicating that the transport is stereoselective. Unlike the human erythrocyte-type glucose transport system, D-ribose had a somewhat greater apparent affinity for the Sf9 cell transporter than D-xylose. It is therefore concluded that Sf9 cells contain an endogenous sugar transport activity that in some aspects resembled the human erythrocyte-type counterpart, although the Sf9 and human transport systems do differ in their affinity for cytochalasin B.

• 한국미생물·생명공학회지
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• 제38권1호
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• pp.7-12
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• 2010

S. cerevisias의 치명적인 약점인 xylose 또는 arabinose가 상당부분을 차지하는 hemicellulose 기수분해물인 오탄당 발효의 극히 낮은 효율은 유전자 변형 및 대사적 흐름을 조절하여 세포 내로의 오탄당 섭취 및 활용 증가를 위한 연구가 꾸준히 진행되고 있다. S. cerevisie에서 오탄당은 육탄당보다 1-2 배 낮은 친화력을 가지고 있어, 오탄당 운반은 이를 이용한 바이오에탄올 발효에 있어서 중요한 초기 조절 단계이다. 오탄당 이용가능 S. cereivsiae에서 오탄당 운반기의 발현 관련 소수의 연구가 보고되고는 있으나 아직까지 눈에 띠는 효율 증기눈 보교되지 않았다. 최근 보고된 S. cerevisiae에서 C. intermeda 유래의 glucose/xylose의 확산을 용이하게 하는 운반기와 공동수송기의 이종발현이 처음으로 활성화 되었음이 보고 되었다. 따라서 그러므로 높은 친화력의 xylose 운반기의 발현은 이미 xylose로 부터 바이오에탄올 발효공정은 최적화되어 있지만 여전히 몇 가지 제한 요소들을 가지고 있는 S. cerevisiae 균주들의 xylsoe 발효공정 효율 향상에 큰 기여를 할 수 있을 것으로 기대된다.

• Journal of Veterinary Science
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• 제23권5호
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• pp.76.1-76.14
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• 2022

Background: Clinical dexamethasone (DEX) treatment or stress in bovines results in extensive physiological changes with prominent hyperglycemia and neutrophils dysfunction. Objectives: To elucidate the effects of DEX treatment in vivo on cellular energy status and the underlying mechanism in circulating neutrophils. Methods: We selected eight-month-old male bovines and injected DEX for 3 consecutive days (1 time/d). The levels of glucose, total protein (TP), total cholesterol (TC), and the proinflammatory cytokines interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α in blood were examined, and we then detected glycogen and adenosine triphosphate (ATP) content, phosphofructosekinase-1 (PFK1) and glucose-6-phosphate dehydrogenase (G6PDH) activity, glucose transporter (GLUT)1, GLUT4, sodium/glucose cotransporter (SGLT)1 and citrate synthase (CS) protein expression and autophagy levels in circulating neutrophils. Results: DEX injection markedly increased blood glucose, TP and TC levels, the Ca²⁺/P⁵⁺ ratio and the neutrophil/lymphocyte ratio and significantly decreased blood IL-1β, IL-6 and TNF-α levels. Particularly in neutrophils, DEX injection inhibited p65-NFκB activation and elevated glycogen and ATP contents and SGLT1, GLUT1 and GR expression while inhibiting PFK1 activity, enhancing G6PDH activity and CS expression and lowering cell autophagy levels. Conclusions: DEX induced neutrophils glucose uptake by enhancing SGLT1 and GLUT1 expression and the transformation of energy metabolism from glycolysis to pentose phosphate pathway (PPP)-tricarboxylic acid (TCA) cycle. This finding gives us a new perspective on deeper understanding of clinical anti-inflammatory effects of DEX on bovine.