• Parasites, Hosts and Diseases
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• 제57권3호
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• pp.249-256
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• 2019

Steroids are commonly used in patients with eosinophilic meningitis caused by A. cantonensis infections. The mechanism steroids act on eosinophilic meningitis remains unclear. In this mouse experiments, expressions of 14-3-3 isoform ${\beta}$ and ${\gamma}$ proteins significantly increased in the CSF 2-3 weeks after the infection, but not increasedin the dexamethasone-treated group. Expression of 14-3-3 ${\beta}$, ${\gamma}$, ${\varepsilon}$, and ${\theta}$ isoforms increased in brain meninges over the 3-week period after infection and decreased due to dexamethasone treatment. In conclusion, administration of dexamethasone in mice with eosinophilic meningitis decreased expressions of 14-3-3 isoform proteins in the CSF and in brain meninges.

• 한국식품영양학회지
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• 제24권2호
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• pp.258-267
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• 2011

14-3-3 is a highly conserved, ubiquitously expressed protein family. It associates with diverse cellular proteins through its specific phosphoserine/phosphothreonine-binding activity and thus contributes to the regulation of crucial cellular processes such as metabolism, signal transduction, cell-cycle control, apoptosis, protein trafficking, transcription and stress responses. This study aims to determine changes in levels of 14-3-3 isoforms and 14-3-3 - associated proteins in Helicobacter pylori(H. pylori)-infected gastric epithelial AGS cells. AGS cells were stimulated with H. pylori(NCTC 11637) at the ratio of 300:1(bacterium:cell). Western blot analysis revealed that 14-3-3 $\sigma$ was elevated at 3 hr after H. pylori treatment. Other isoforms were not significantly affected by H. pylori infection. Using immunoprecipitation to 14-3-3 $\sigma$, followed by proteomic analysis, we found that S phase kinase associated protein isoform 2 bound to 14-3-3 $\sigma$ has increased. In contrast, three proteins (DEAD-box polypeptide 3, heterogeneous nuclear ribonucleoprotein H2 and WD repeat-containing protein isoform 1) bound to 14-3-3 decreased by H. pylori infection. Our results suggest that 14-3-3 may play an important regulatory role in H. pylori-induced signal transduction in gastric epithelial cells.

• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1253-1259
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• 2012

The 14-3-3 proteins are highly conserved, ubiquitous molecules involved in a variety of biologic phenomena, such as cell cycle control, and apoptosis. However, their expression in cholangiocarcinoma has not been previously characterized. In this paper, immunohistochemistry using specific anti-14-3-3 monoclonal antibodies was performed on formalin-fixed;, paraffin embedded archival tissue from 86 patients of cholangiocarcinoma. We also examined the correlation between expression and survival rate and clinicopathologic factors such as tumor location, tumor size, pathologic differentiation, lymphatic permeation, lymph node metastasis, and tumor stage. Positive 14-3-3 proteins expression was observed for 6 isoforms (${\beta}$, ${\sigma}$, ${\gamma}$, ${\theta}$, ${\delta}$, ${\eta}$) of these proteins in 86 patients of cholangiocarcinoma. ${\beta}$ and ${\sigma}$ isoform immunoreactivity was correlated with lymph node metastasis, tumor stage and patients' survival rate. In addition, ${\delta}$ isoform immunoreactivity showed trends with tumor location, tumor size, pathologic differentiation and tumor stage, while the ${\theta}$ isoform was correlated with pathologic differentiation. These results indicated that upregulated expression of some isoforms of 14-3-3 may be a common mechanism for evading apoptosis in cholangiocarcinoma, so that targeting 14-3-3 may be a novel promising strategy for the treatment of this tumor.

• The Korean Journal of Physiology and Pharmacology
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• 제14권1호
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• pp.45-49
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• 2010

R-type $Ca_v2.3$ high voltage-activated $Ca^{2+}$ channels in peripheral sensory neurons contribute to pain transmission. Recently we have demonstrated that, among the six $Ca_v2.3$ isoforms ($Ca_v2.3a{\sim}Ca_v2.3e$), the $Ca_v2.3e$ isoform is primarily expressed in trigeminal ganglion (TG) nociceptive neurons. In the present study, we further investigated expression patterns of $Ca_v2.3$ isoforms in the dorsal root ganglion (DRG) neurons. As in TG neurons, whole tissue RT-PCR analyses revealed the presence of two isoforms, $Ca_v2.3a$ and $Ca_v2.3e$, in DRG neurons. Single-cell RT-PCR detected the expression of $Ca_v2.3e$ mRNA in 20% (n=14/70) of DRG neurons, relative to $Ca_v2.3a$ expression in 2.8% (n=2/70) of DRG neurons. $Ca_v2.3e$ mRNA was mainly detected in small-sized neurons (n=12/14), but in only a few medium-sized neurons (n=2/14) and not in large-sized neurons, indicating the prominence of $Ca_v2.3e$ in nociceptive DRG neurons. Moreover, $Ca_v2.3e$ was preferentially expressed in tyrosine-kinase A (trkA)-positive, isolectin B4 (IB4)-negative and transient receptor potential vanilloid 1 (TRPV1)-positive neurons. These results suggest that $Ca_v2.3e$ may be the main R-type $Ca^{2+}$ channel isoform in nociceptive DRG neurons and thereby a potential target for pain treatment, not only in the trigeminal system but also in the spinal system.

• 한국발생생물학회지:발생과생식
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• 제2권2호
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• pp.165-171
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• 1998

p62는 임파구에 특이적으로 발현하는 단백질 티로신 키나제인 p56$^{lck}$의 SH2 doamin과 결합하는 세포질 단백질로서 두 단백질의 결합에는 지금까지 알려진 바와 다르게 인산화된 티로신이 필요없다. p62는 기능이 다른 여러 조직에서 공통적으로 발현되며 유비퀴틴, 단백질 키나제 C 이성질체 둥 다양한 단백질과 결합하는 것이 알려져 있다. 이와 같은 현상으로 p62가 다양한 생물학적 기능을 수행할 수 있음을 예측할 수 있으나 그 자세한 기작은 잘 알려져 있지 않다. 본 연구에서는 p62가 T-세포에 특이적으로 발현하는 14-3-3 $ au$ 이성질체와 결합하는 것을 확인하였으며, p62를 인위적으로 T-세포에 다량으로 발현시키면 세포예정사 (apoptosis)의 시작이 지연되는 현상을 조사하였다. 이때 세포사멸과정에서 전형적으로 나타나는 DNA 절단현상 (DNA fragmentaion)과 poly (ADP-ribose) polymerase의 분해가 지연됨을 알 수 있었다. 최근 14-3-3 단백질이 임파구에서 세포예정사를 촉진시키는 기능을 가진 Bad와 결합함으로써 세포의 생존 신호 전달에 중요한 역할을 한다는 것이 보고된 바 있다. 따라서 본 연구의 결과는 T-세포의 활성으로 일어나는 사멸예정사 과정 중에 p62와 14-3-3 단백질에 의해 수행되는 조절 기작이 있음을 시사하고 있다.다.

• IMMUNE NETWORK
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• 제14권3호
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• pp.123-127
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• 2014

Interleukin-32 (IL-32) is a cytokine inducing crucial inflammatory cytokines such as tumor necrosis factor-${\alpha}(TNF{\alpha})$ and IL-6 and its expression is elevated in various inflammatory autoimmune diseases, certain cancers, as well as viral infections. IL-32 gene was first cloned from activated T cells, however IL-32 expression was also found in other immune cells and non-immune cells. IL-32 gene was identified in most mammals except rodents. It is transcribed as multiple-spliced variants in the absence of a specific activity of each isoform. IL-32 has been studied mostly in clinical fields such as infection, autoimmune, cancer, vascular disease, and pulmonary diseases. It is difficult to investigate the precise role of IL-32 in vivo due to the absence of IL-32 gene in mouse. The lack of mouse IL-32 gene restricts in vivo studies and restrains further development of IL-32 research in clinical applications although IL-32 new cytokine getting a spotlight as an immune regulatory molecule processing important roles in autoimmune, infection, and cancer. In this review, we discuss the regulation and function of IL-32 in inflammatory bowel diseases and rheumatoid arthritis.

• The Korean Journal of Physiology and Pharmacology
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• 제24권1호
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• pp.81-88
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• 2020

Regulator of calcineurin 1 (RCAN1) can be induced by an intracellular calcium increase and oxidative stress, which are characteristic features of temporal lobe epilepsy. Thus, we investigated the spatiotemporal expression and cellular localization of RCAN1 protein and mRNA in the mouse hippocampus after pilocarpine-induced status epilepticus (SE). Male C57BL/6 mice were given pilocarpine hydrochloride (280 mg/kg, i.p.) and allowed to develop 2 h of SE. Then the animals were given diazepam (10 mg/kg, i.p.) to stop the seizures and sacrificed at 1, 3, 7, 14, or 28 day after SE. Cresyl violet staining showed that pilocarpine-induced SE resulted in cell death in the CA1 and CA3 subfields of the hippocampus from 3 day after SE. RCAN1 immunoreactivity showed that RCAN1 was mainly expressed in neurons in the shammanipulated hippocampi. At 1 day after SE, RCAN1 expression became detected in hippocampal neuropils. However, RCAN1 signals were markedly enhanced in cells with stellate morphology at 3 and 7 day after SE, which were confirmed to be reactive astrocytes, but not microglia by double immunofluorescence. In addition, realtime reverse transcriptase-polymerase chain reaction showed a significant upregulation of RCAN1 isoform 4 (RCAN1-4) mRNA in the SE-induced hippocampi. Finally, in situ hybridization with immunohistochemistry revealed astrocytic expression of RCAN1-4 after SE. These results demonstrate astrocytic upregulation of RCAN1 and RCAN1-4 in the mouse hippocampus in the acute and subacute phases of epileptogenesis, providing foundational information for the potential role of RCAN1 in reactive astrocytes during epileptogenesis.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권3호
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• pp.200-208
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• 2006

Amino acids are required for protein synthesis and energy sources in all living cells. The amino acid transport system L is a major nutrient transport system that is responsible for $Na^+$-independent transport of neutral amino acids including several essential amino acids. In malignant tumors, the L-type amino acid transporter 1 (LAT1), the first isoform of system L, is highly expressed to support tumor cell growth. In the present study, the expression and functional characterization of amino acid transport system L were, therefore, investigated in Saos2 human osteogenic sarcoma cells. RT-PCR and western blot analyses have revealed that the Saos2 cells expressed the LAT1 and the L-type amino acid transporter 2 (LAT2), the second isoform of system L, together with their associating protein heavy chain of 4F2 antigen (4F2hc) in the plasma membrane, but the expression of LAT2 was very weak. The uptakes of [${14}^C$]L-leucine by Saos2 cells were $Na^+$-independent and were completely inhibited by the system L selective inhibitor, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). The affinity of [${14}^C$]L-leucine uptake and the inhibition profiles of [${14}^C$]L-leucine uptake by various amino acids in the Saos2 cells were comparable with those for the LAT1 expressed in Xenopus oocytes. The majority of [${14}^C$]L-leucine uptake is, therefore, mediated by LAT1 in the Saos2 cells. These results suggest that the transports of neutral amino acids including several essential amino acids into Saos2 human osteogenic sarcoma cells are for the most part mediated by LAT1. Therefore, the Saos2 human osteogenic sarcoma cells are excellent tools for examine the properties of LAT1. Moreover, the specific inhibition of LAT1 in tumor cells might be a new rationale for anti-tumor therapy.

• 치위생과학회지
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• 제13권3호
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• pp.296-303
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• 2013

LOX는 결합조직의 세포외기질에 존재하여 교원섬유와 탄성섬유의 교차결합을 촉진시키는 핵심적인 효소이다. 뼈세포에서 세포분화 및 세포이동과 관련된 LOX의 역할은 보고되었으나, 상아모세포 분화에서 LOX에 대한 직접적인 효과는 거의 알려지지 않았다. 이 연구에서는 인간치수세포에서 상아모세의 분화과정 동안 LOX의 역할에 대해 실험하였다. 치수세포에 저 혈청 분화유도 배지를 처리하여 0, 1, 3, 7, 14일 동안 배양하였다. 세포성장은 세포계수, 분화와 관련된 유전자들은 RT-PCR, 광물화는 alrizarin red S 염색으로 각각 실험하였다. LOX 유전자 발현은 RT-PCR로 측정하였고, LOX 효소활성은 고감도 형광분석을 시행하였다. 1. 세포성장은 저 혈청 분화유도 배지에서 시간 의존적으로 증가하였다. 2. 분화표지자인 ALP, OPN, OCN, DMP1, DSPP는 시간 의존적으로 발현되었으나 초기, 중기, 말기에 대한 차별화는 이루어지지 않았다. 3. 광물화는 3일부터 관찰되기 시작하여 14일까지 증가되었다. 4. LOX와 LOXL은 mRNA 발현이 7일까지 증가되었고, 14일에서 큰 감소를 보였다. 하지만, LOXL3와 LOXL4 mRNA는 낮은 발현을 보였고, LOXL2 mRNA는 발현되지않았다. 5. Collagen type I mRNA는 7일까지 증가하다가 14일에서 의미있게 감소를 보였고, collagen type IV는 낮은 mRNA 발현을 보였다. 6. LOX 효소 활성은 분화유도기간 중 7일에서 가장 많은 발현을 보였고, 교원질 1형 기질이 교원질 IV형 기질보다 두드러진 발현을 보였다. 이상의 결과를 종합해 볼 때, LOX isoform의 발현과 LOX 효소 활성은 인간치수세포에서 상아모세포 분화과정 동안 중요한 조절자로 사료된다.

• Food Science and Biotechnology
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• 제14권5호
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• pp.639-644
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• 2005

The aim of this study was to investigate the effect of the myosin light chain (MLC) isoforms on the muscle fiber characteristics and meat quality traits in porcine longissimus muscle. Pale, soft, exudative (PSE) samples had a lower content of essential light chain (ELC) 1S isoforms and a higher proportion of the fiber type IIB than the reddish-pink, firm, non-exudative (RFN) samples. These compositions suggest that the PSE pork has a higher glycolytic and a lower oxidative capacity than the RFN pork. Therefore, these characteristics of PSE pork might affect the metabolic rate and meat quality traits, including protein solubility. In addition, the indicator traits of the postmortem metabolic rate were related to the ELC 1F/3F ratio ($pH_{45\;min}$: r = -0.43, P < 0.001; R-value: r = 0.53, P < 0.001). These results suggest that the MLC isoform composition can affect the postmortem metabolic rate and meat quality traits.