• Molecules and Cells
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• v.27 no.5
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• pp.577-582
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• 2009

The development of a high-throughput functional genomic screening provides a novel and expeditious approach in identifying critical genes involved in specific biological processes. Here we describe a cell-based cDNA screening system to identify the transcription activators of BiP, an endoplasmic reticulum (ER) chaperone protein. BiP promoter contains the ER stress element which is commonly present in the genes involved in unfolded protein response (UPR) that regulates protein secretion in cells. Therefore, the positive regulators of BiP may also be utilized to improve the recombinant protein production through modulation of UPR. Four BiP activators, including human UDP-glucose:glycoprotein glucosyltransferase 1 (HUGT1), are identified by the cDNA screening. Overexpression of HUGT1 leads to a significant increase in the production of recombinant erythropoietin, interferon ${\gamma}$, and monoclonal antibody in HEK293 cells. Our results demonstrate that the cDNA screening for BiP activators may be effective to identify the novel BiP regulators and HUGT1 may serve as an ideal target gene for improving the recombinant protein production in mammalian cells.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.25 no.2
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• pp.126-133
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• 2015

연구목적: 이 연구는 노동부의 보건진단 명령에 의해 유리섬유 강화플라스틱(FRP)을 이용한 이중벽 탱크 제조 사업장의 적층 공정 근로자들을 대상으로 스티렌 노출 특성을 평가하기 위해 수행되었다. 연구방법: 스티렌의 주요발생원 파악을 위해 불포화 폴리에스테르 수지(UPR), 경화제, 조색제, 세척액 등의 원료 내 스티렌 함유량을 가스크로마토그래피 질량분석기(GC-MS)를 이용하여 분석하였다. FRP 적층 공정에 근무하는 작업자들을 대상으로 NIOSH 1501 공정시험법에 의해 공기 중 스티렌 노출 농도에 대한 개인노출 평가를 실시하였고, 생물학적 노출 지표로 뇨 중 만델릭산을 채취한 후 고성능액체크로마토그래피(HPLC)로 분석하였다. 또한 각 직무 특성과 단위작업 중심으로 스티렌에 대한 단시간 노출평가를 수행하였다. 연구결과: 스티렌의 함유량이 가장 많은 주요 원료는 중량비율로37%의 스티렌이 함유된 UPR이었다. 적층 공정의 FRP분무 작업자와 보조 작업자들 모두 스티렌의 8시간 가중평균 노출기준(20 ppm)을 초과하였다. 단시간 노출평가 결과 FRP분무 작업자의 경우 45.9 ppm에서 86.1 ppm 수준으로 FRP를 사용하지 않는 작업보다 통계적으로 유의하게 높은 수준이었다(P<0.01). 가장 높은 수준의 스티렌에 노출되는 단위작업은 FRP를 이용하여 1차 코팅 하는 작업으로 특별한 관리가 필요하였다. 결론: 보건진단을 위해 실시한 이중벽 탱크 제조 사업장의FRP 적층 공정 작업자의 스티렌 노출수준은 노출기준을 초과할 정도로 높은 수준이었다. 그러나 탱크를 천장에 매달고 돌리면서 적층작업을 수행하기 때문에 적절한 국소환기 시스템을 구축하는데 어려움이 있다. 따라서 적절한 방독마스크 착용으로 스티렌 노출 예방이 필요하였다.

• Membrane and Water Treatment
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• v.2 no.4
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• pp.251-266
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• 2011

This work deals with the use of High Resolution Scanning Electron Microscopy (HRSEM) to verify ultrafiltration membrane selectivity at the end of the production line as well as membrane ageing. The first part of this work is focused on new membranes. It is shown that it is better to use sputtering metallization than vacuum deposition, as this latter technique entails thermal damage to the skin layer. Moreover, the impact of the metallization layer on the determination of the membrane pore size is studied and it is observed that no impact of the metallization step can be clearly defined for a metallization layer ranging from 3 to 12 nm. For example, an average pore size of 16.9 nm and a recovery rate of 6.5 % are observed for a 150 kDa cellulose acetate membrane. These results are in agreement with those given by the manufacturer: pore size ranging from 10 to 15 nm and recovery rate ranging from 5 to 10 %. The second part of this work focuses on the study of membrane ageing. A PVDF hollow fibre membrane is studied. It is shown that a 65 % decrease in the permeate flux can be linked to a decrease in the number of pores at the surface of the membrane and a decrease in the recovery rate. In conclusion, a mapping of the pores is performed for several new hollow fibre membranes used to produce drinking water, made of different materials, with different geometries and molecular weight cut-off. These results provide reference data that will help better understand the phenomena of membrane fouling and membrane ageing.

• Journal of Life Science
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• v.17 no.6 s.86
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• pp.847-858
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• 2007

The endoplasmic reticulum (ER) is an important intracellular organelle for folding and maturation of newly synthesized transmembrane and secretory proteins. The ER provides stringent quality control systems to ensure that only correctly folded proteins exit the ER and unfolded or misfolded proteins are retained and ultimately degraded. The ER has evolved stress response both signaling pathways the unfolded protein response (UPR) to cope with the accmulation of unfolded or misfolded proteins and ER overload response (EOR). Accumulating evidence suggests that, in addition to responsibility for protein processing, ER is also an important signaling compartment and a sensor of cellular stress. In this respect, production of bio-functional recombinant-proteins requires efficient functioning of the ER secretory pathway in host cells. This review briefly summarizes our understanding of the ER signaling developed in the recent years to help of the secretion capacities of recombinant cells.

• Journal of Veterinary Science
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• v.25 no.2
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• pp.21.1-21.15
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• 2024

Background: Peste des petits ruminants (PPR) is a contagious and fatal disease of sheep and goats. PPR virus (PPRV) infection induces endoplasmic reticulum (ER) stress-mediated unfolded protein response (UPR). The activation of UPR signaling pathways and their impact on apoptosis and virus replication remains controversial. Objectives: To investigate the role of PPRV-induced ER stress and the IRE1-XBP1 and IRE1-JNK pathways and their impact on apoptosis and virus replication. Methods: The cell viability and virus replication were assessed by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, immunofluorescence assay, and Western blot. The expression of ER stress biomarker GRP78, IRE1, and its downstream molecules, PPRV-N protein, and apoptosis-related proteins was detected by Western blot and quantitative reverse transcription-polymerase chain reaction, respectively. 4-Phenylbutyric acid (4-PBA) and STF-083010 were respectively used to inhibit ER stress and IRE1 signaling pathway. Results: The expression of GRP78, IRE1α, p-IRE1α, XBP1s, JNK, p-JNK, caspase-3, caspase-9, Bax and PPRV-N were significantly up-regulated in PPRV-infected cells, the expression of Bcl-2 was significantly down-regulated. Due to 4-PBA treatment, the expression of GRP78, p-IRE1α, XBP1s, p-JNK, caspase-3, caspase-9, Bax, and PPRV-N were significantly downregulated, the expression of Bcl-2 was significantly up-regulated. Moreover, in PPRV-infected cells, the expression of p-IRE1α, p-JNK, Bax, and PPRV-N was significantly decreased, and the expression of Bcl-2 was increased in the presence of STF-083010. Conclusions: PPRV infection induces ER stress and IRE1 activation, resulting in apoptosis and enhancement of virus replication through IRE1-XBP1s and IRE1-JNK pathways.

• Biomolecules & Therapeutics
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• v.32 no.3
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• pp.341-348
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• 2024

Endoplasmic reticulum (ER) stress plays a crucial role in liver diseases, affecting various types of hepatic cells. While studies have focused on the link between ER stress and hepatocytes as well as hepatic stellate cells (HSCs), the precise involvement of hepatic macrophages in ER stress-induced liver injury remains poorly understood. Here, we examined the effects of ER stress on hepatic macrophages and their role in liver injury. Acute ER stress led to the accumulation and activation of hepatic macrophages, which preceded hepatocyte apoptosis. Notably, macrophage depletion mitigated liver injury induced by ER stress, underscoring their detrimental role. Mechanistic studies revealed that ER stress stimulates macrophages predominantly via the PERK signaling pathway, regardless of its canonical substrate ATF4. hnRNPA1 has been identified as a crucial mediator of PERK-driven macrophage activation, as the overexpression of hnRNPA1 effectively reduced ER stress and suppressed pro-inflammatory activation. We observed that hnRNPA1 interacts with mRNAs that encode UPR-related proteins, indicating its role in the regulation of ER stress response in macrophages. These findings illuminate the cell type-specific responses to ER stress and the significance of hepatic macrophages in ER stress-induced liver injury. Collectively, the PERK-hnRNPA1 axis has been discovered as a molecular mechanism for macrophage activation, presenting prospective therapeutic targets for inflammatory hepatic diseases such as acute liver injury.

• Journal of Life Science
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• v.24 no.10
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• pp.1039-1045
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• 2014

This study analyzed whether human chorionic gonadotropin (hCG) induces ER stress via the IRE/XBP1 pathway in mouse Leydig tumor (mLTC-1) cells. In a previous study, we demonstrated that the unfolding protein response (UPR) plays an important role in the expression of steroidogenic enzymes by modulating the ATF6 pathway, as well as ER stress-mediated apoptosis in hCG-stimulated Leydig cells. Although UPR signaling has been reported to regulate the IRE1/XBP1 pathway, it is not known whether hCG-induced ER stress in Leydig cells can activate the pathway. To investigate the activation of the IRE1/XBP1 pathway in mLTC-1 cells after hCG treatment, we performed a Western blot analysis to detect the phospho-IRE1 protein and an RT-PCR analysis to validate splicing of XBP1 mRNA. We used ER stress-activated indicator (ERAI) constructs for monitoring the activity of IRE1 and then analyzed by fluorescence microscopy and flow cytometry. The expression levels of the phospho-IRE1 protein markedly increased in response to the hCG treatment. In the mLTC-1 cells transfected with an F-XBP1-venus/F-$XBP1{\Delta}DBD$-venus construct, the hCG treatment led to the appearance of green fluorescent cells and detectable fluorescence in the nucleus and cytosol, respectively. In addition, splicing of XBP1 mRNA significantly increased after the hCG treatment. Taken together, these results indicate that hCG-induced ER stress leads to activation of the IRE1/XBP pathway in Leydig cells.

• The Korean Journal of Air & Space Law and Policy
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• v.20 no.1
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• pp.255-272
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• 2005

This study was conducted to present the adaquate guidelines necessary in decision making processes regarding air traffic management as well as airspace management in the national airspace system. It encompasses the overall advanced air traffic management initiatives, domestic air traffic congestion and resolution method as well.

• Biomedical Science Letters
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• v.13 no.2
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• pp.157-160
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• 2007

Endoplasmic reticulum (ER) plays formation of disulfide bonds and proper folding of secretory proteins. Cellular responses to ER stress enhances the stress-activated kinase pathway and the induces a lot of immediate-early genes. Among of them, the early growth response-1 (Egr-1), a transcription factor, which plays an important role in cell growth, development, differentiation, apoptosis and various types of injury. For that reason, we have tested the expression of Egr-1 against ER stress inducible drugs (tunicamycin, DTT, A23187 and BFA) to understand what kind of aspect occurred by ER stresses.

• Biomedical Science Letters
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• v.16 no.1
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• pp.65-67
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• 2010

The endoplasmic reticulum (ER) is a multifunctional intercellular organelle in which several posttranslational modification steps occurred such as protein folding, lipid biosynthesis, calcium storage and release. Perturbations that disrupt ER homeostasis lead to the misfolding of proteins in the ER lumen and up-regulation of ER signaling pathway called the unfolded protein response (UPR). Here, we have demonstrated that ageing changes the expression of ER chaperone and associated ER membrane kinases of IRE1, ATF6 and PERK.