• Title/Summary/Keyword: 14-3-3 isoforms

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The Clinicopathological and Prognostic Impact of 14-3-3 Protein Isoforms Expression in Human Cholangiocarcinoma by Immunohistochemistry

  • Wu, Qiao;Liu, Chang-Zheng;Tao, Lian-Yuan;Yu, Lan;Liu, Wei;Chen, Song-Sen;He, Xiao-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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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.

14-3-3-Associated Proteins in Helicobacter pylori-Infected Gastric Epithelial Cells (Helicobacterpylori에 감염된 위상피세포에서 14-3-3 결합 단백질의 변화)

  • Chung, Hae-Yun
    • The Korean Journal of Food And Nutrition
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    • v.24 no.2
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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.

Dexamethasone Downregulates Expressions of 14-3-3β and γ-Isoforms in Mice with Eosinophilic Meningitis Caused by Angiostrongylus cantonensis Infection

  • Tsai, Hung-Chin;Chen, Yu-Hsin;Yen, Chuan-Min;Chung, Li-Yu;Wann, Shue-Ren;Lee, Susan Shin-Jung;Chen, Yao-Shen
    • Parasites, Hosts and Diseases
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    • v.57 no.3
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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.

Relationship Among Apolopoprotein E Phenotypes, Dietary Fat, Serum Lipoprotein Concentrations and Erythrocyte Membrane Fatty Acid Composition in Young Healthy Women (건강한 젊은 여성의 Apolipoprotein E 의 다형성 , 식이 지방 섭취, 혈청 지단백 농도와 적혈구 막의 지방산 조성과의 관계)

  • 박선민
    • Journal of Nutrition and Health
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    • v.30 no.8
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    • pp.936-951
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    • 1997
  • As fat consumption has increased in Korea , serum cholesterol concentrations have risen and the prevalence of atherosclerosis increased. The aim of the present study was to investigate the association among apolipoprotein E (Apo E) phenotypes, dietary fat consumption, plasma lipoprotein levels and fatty acid compositions of red blood cells in young healthy women. Seventy-one percent of participants had Apo E3/3, 14.9 percent had ApoE3/2 , 11.8% had Apo E4/3 and 1.2% had Apo E4.2. The subjects daily consumed approximately 1760kcal containing 29 energy % of fat. The ratio of polyunsaturated fat to saturated fat in a diet was 0. 73 . There were no significant differences of nutrient intakes according to Apo E phenotypes. Total cholesterol concentrations of subjects averaged approximately 160mg/dl , but 12 percent of them had over 200mg/dl, which is higher than the range recommended by the National Cholesterol Deucation Program. Notably, most subjects with 210mg/dl had Apo E4 isoforms. Subjects with Apo E4 isoforms had significantly higher total and LDL-cholesterol concentrations than those with Apo E3/2. Also subjects carrying Apo E4 isoforms tended to accumulated more fat in the body. Their BMI , WHR and arm fat area appeared to be how can arm fat area be greater no you mean grater, please check work vsage than subjects with Apo E3 isoforms, but not significantly. Fat intakes slightly influenced serum cholesterol levels. Myristic aicd intakes were positively correlated to serum total and LDL-choelsterol levels. Polyunsaturated fatty acid intakes were negatively correlated to serum total choelsterollevels. The results of 소냐 study suggest that , people with Apo E4 isoforms need to prevent the raising of serum total and LDL -cholesterol concentrations by reducing calorie and fat intakes and by maintaining a normal weight.

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R-type Calcium Channel Isoform in Rat Dorsal Root Ganglion Neurons

  • Fang, Zhi;Hwang, Jae-Hong;Kim, Joong-Soo;Jung, Sung-Jun;Oh, Seog-Bae
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.1
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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.

Analysis of Myosin Heavy Chain Isoforms from Longissimus Thoracis Muscle of Hanwoo Steer by Electrophoresis and LC-MS/MS

  • Kim, Gap-Don
    • Food Science of Animal Resources
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    • v.34 no.5
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    • pp.656-664
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    • 2014
  • The purpose of this study was to analyze myosin heavy chain (MHC) isoforms in bovine longissimus thoracis (LT) muscle by liquid chromatography (LC) and mass spectrometry (MS). LT muscles taken from Hanwoo (Korean native cattle) steer (n=3) used to separate myosin bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The peptide queries were obtained from the myosin bands by LC-MS/MS analysis following in-gel digestion with trypsin. A total of 33 and 43 queries were identified as common and unique peptides, respectively, of MHC isoforms (individual ions scores >43 indicate identity or extensive homology, p<0.05). MHC-1 (IIx), -2 (IIa), -4 (IIb), and -7 (slow/I) were identified based on the Mowse score (5118, 3951, 2526, and 2541 for MHC-1, -2, -4, and -7, respectively). However, more analysis is needed to confirm the expression of MHC-4 in bovine LT muscle because any query identified as a unique peptide of MHC-4 was not found. The queries that were identified as unique peptides could be used as peptide markers to confirm MHC-1 (14 queries), -2 (8 queries), and -7 (21 queries) in bovine LT muscle; no query identified as a unique peptide of MHC-4 was found. LC-MS/MS analysis is a useful approach to study MHC isoforms at the protein level.

Relationships between Myosin Light Chain Isoforms, Muscle Fiber Characteristics, and Meat Quality Traits in Porcine Longissimus Muscle

  • Choi, Young-Min;Ryu, Youn-Chul;Lee, Sang-Hoon;Kim, Byoung-Chul
    • Food Science and Biotechnology
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    • v.14 no.5
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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.

Discovery of 14-3-3 zeta as a potential biomarker for cardiac hypertrophy

  • Joyeta Mahmud;Hien Thi My Ong;Eda Ates;Hong Seog Seo;Min-Jung Kang
    • BMB Reports
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    • v.56 no.6
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    • pp.341-346
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    • 2023
  • Acute myocardial infarction (AMI) is a multifaceted syndrome influenced by the functions of various extrinsic and intrinsic pathways and pathological processes, which can be detected in circulation using biomarkers. In this study, we investigated the secretome protein profile of induced-hypertrophy cardiomyocytes to identify next-generation biomarkers for AMI diagnosis and management. Hypertrophy was successfully induced in immortalized human cardiomyocytes (T0445) by 200 nM ET-1 and 1 μM Ang II. The protein profiles of hypertrophied cardiomyocyte secretomes were analyzed by nano-liquid chromatography with tandem mass spectrometry and differentially expressed proteins that have been identified by Ingenuity Pathway Analysis. The levels of 32 proteins increased significantly (>1.4 fold), whereas 17 proteins (<0.5 fold) showed a rapid decrease in expression. Proteomic analysis showed significant upregulation of six 14-3-3 protein isoforms in hypertrophied cardiomyocytes compared to those in control cells. Multi-reaction monitoring results of human plasma samples showed that 14-3-3 protein-zeta levels were significantly elevated in patients with AMI compared to those of healthy controls. These findings elucidated the role of 14-3-3 protein-zeta in cardiac hypertrophy and cardiovascular disorders and demonstrated its potential as a novel biomarker and therapeutic strategy.

Effects of PCB Congeners in Rodent Neuronal Cells in Culture : Effects of Chitosan (PCB 이성질체가 설치류 신경세포에 미치는 영향: 키토산의 효과)

  • Kim, Sun-Young;Lee, Hyun-Gyo
    • Environmental Analysis Health and Toxicology
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    • v.22 no.3
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    • pp.279-285
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    • 2007
  • The present study attempted to analyze the mechanism of PCB-induced neurotoxicity with respect to the PKC signaling. Since the developing neuron is particularly sensitive to PCB-induced neurotoxicity, we isolated cerebellar granule cells derived from 7-day old SD rats and grew cells in culture for additional 7 days to mimic PND-14 conditions. Only non-coplanar PCBs at a high dose showed a significant increase of total PKC activity at $[^3H]PDBu$ binding assay, indicating that non-coplanar PCBs are more neuroactive than coplanar PCBs in neuronal cells. PKC isoforms were immunoblotted with respective monoclonal antibodies. PKC-alpha and-epsilon were activated with non-coplanar PCB exposure. The result suggests that coplanar PCBs have a PKC pathway different from non-coplanar PCBs. Activation of PKC with exposure was dampened with treatment of high molecular weight of chitosan. Chilean (M.W. > 1,000 kDa) inhibited the total activity of PKC induced by the non-coplanar PCBs. Translocation of PKC isoforms was also inhibited by the high molecular weight of chitosan. The study demonstrated that non-coplanar PCBs are more potent neurotoxic congeners than coplanar PCBs and the alteration of PKC activities by PCB exposure can be blocked with the treatment of chitosan. The results suggest a potential use of chitosan as a means of nutritional intervention to prevent the harmful effects of pollutant-derived diseases.

Expression Profiling of Connexins in the Efferent Ductules of Male Rats During Postnatal Development (생후 발달과정동안 숫 백서의 Efferent Ductules에서 Connexins 발현 양상 연구)

  • Lee, K.H.;Choi, I.H.;Chung, C.S.;Chang, J.S.;Kim, J.Y.
    • Journal of Animal Science and Technology
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    • v.49 no.1
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    • pp.15-24
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    • 2007
  • The purpose of this study was to evaluate the presence and differential expression of connexin isoforms in the efferent ductules (ED) of male rat reproductive tract during postnatal development. Total RNA was isolated from the ED collected from rats at 1 week, 2 weeks, 1 month, 3 months, and 6 months of ages. Expression of six connexin mRNAs of 14 isoforms tested was detected by semi-quantitative RT-PCR. Fluctuation of mRNA levels of connexins 26 and 30 was found according to ages. A significant decrease of connexin 31.1 mRNA level was observed in the ED at 1 month of age. The highest levels of connexin 37 and 45 mRNAs were detected in the ED of early developmental period, while the expression of connexin 43 was the lowest at 1 week of age. The present study demonstrates differential regulation on expression of connexin isoforms in the rat ED in age-dependent manner.