• Title/Summary/Keyword: protein tissues

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Cloning and Expression Analysis of a Novel Mouse Zinc Finger Protein Gene Znf313 Abundantly Expressed in Testis

  • Li, Na;Sun, Huaqin;Wu, Qiaqing;Tao, Dachang;Zhang, Sizhong;Ma, Yongxin
    • BMB Reports
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    • v.40 no.2
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    • pp.270-276
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    • 2007
  • We have cloned a novel mouse zinc finger protein gene Znf313 by rapid amplification of cDNA ends (RACE) according to the homologue of human ZNF313 gene. The cDNA is 2,163 base pairs (bp) in length and encodes a 229 amino acids (aa) protein with a $C_3HC_4$ ring finger domain and three $C_2H_2$ domains. 89% and 93% nucleotide (nt) and aa sequence identity is observed with its human homologue. Revealed by Northern blot and RT-PCR, full mRNA consists of 2.16 kb and widely expresses in tissues as a single transcript, most abundantly in heart, liver, kidney and testis. The expression of Znf313 in testis is detected in all development stages. Western blot analysis also reveals that Znf313 is expressed in the tissues. Immunohistochemical staining and subcellular localization demonstrate that Znf313 is expressed both in the cytoplasm and nucleus whereas predominantly localized in the nucleus. Present data suggests that Znf313 gene might play a fundamental role in gene transcription and regulation in organism and relates to spermatogenesis.

Gene Expression of Heart and Adipocyte Fatty Acid-binding Protein in Chickens by FQ-RT-PCR

  • Tu, Yunjie;Su, Yijun;Wang, Kehua;Zhang, Xueyu;Tong, Haibing;Gao, Yushi
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.8
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    • pp.987-992
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    • 2010
  • This study was to detect the expression of heart fatty acid-binding protein (H-FABP) and adipocyte fatty acid-binding protein (A-FABP) gene mRNA in different tissues of Rugao and Luyuan chickens at 56 d and 120 d by real-time fluorescence quantitative reverse transcription polymerase-chain reaction (FQ-RT-PCR). The primers were designed according to the sequences of HFABP, A-FABP and GAPDH genes in Gallus gallus, which were used as target genes and internal reference gene, respectively. The levels of H-FABP and A-FABP gene expression were detected by SYBR Green I FQ-RT-PCR. The relative H-FABP and A-FABP gene mRNA expression level was calculated with 2-$^{{\Delta}Ct}$. Melting curve analysis showed a single peak of three genes. Intramuscular fat (IMF) content in breast muscle and leg muscle of the two chicken breeds at 120 d was higher than at 56 d. IMF content in breast muscle and leg muscle at 56 d and 120 d in Luyuan was significantly higher than in Rugao, however, abdominal fat of Luyuan was significantly lower than that of Rugao. The relative H-FABP gene mRNA expression level in cardiac muscle was the highest in both chicken breeds. The relative H-FABP and A-FABP gene expression of different tissues in Luyuan was higher than in Rugao. H-FABP gene mRNA expression had a negative effect on IMF of leg and breast muscles, and was significantly negatively correlated with IMF content. The relative A-FABP gene mRNA level in abdominal fat was higher than in liver. The A-FABP gene mRNA was not expressed in leg, breast and cardiac muscles. A-FABP gene mRNA expression level was significantly positively correlated with abdominal fat and had a significant effect on abdominal fat but not IMF content.

Changes in Biochemical Composition of the Digestive Gland of the Female Purple Shell, Rapana venosa, in Relation to the Ovarian Developmental Phases

  • Chung, Ee-Yung;Kim, Sung-Yeon;Park, Kwan-Ha
    • The Korean Journal of Malacology
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    • v.17 no.1
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    • pp.27-33
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    • 2001
  • The Ovarian developmental phases of the reproductive cycle of Rapana venosa can be classified into five successive stages by histological study: early active stage (September to February), late active stage (December to April), ripe stage (March to July), partially spawned stage (May to August), and recovery stage (June to September). To understand the characteristics of nutrient storage and utilization in the digestive gland cells with ovarian developmental phases, we examined the digestive gland - which is the major nutrient supply organ associated with ovarian development of the female purple shell - by biochemical methods. Total protein contents in the digestive gland tissues increased in March (late active stage) and reached the maximum in May (ripe and partially spawned stages), and then their levels sharply decreased in July (partially spawned and recovery stages). Total lipid contents in the digestive gland tissues reached the maximum in January (early active stage). Thereafter, their levels rapidly decreased from May (ripe and partially spawned stages) and reached a minimum in July (partially spawned and recovery stages). The total DNA contents did not significantly change regardless of the different developmental stages of the ovary. However, it was also found from biochemical analysis that changes in total RNA content follow the same seasonal cycling to protein. These results indicate that the digestive gland is an important energy storage and supply organ in purple shells, and that the nutrient contents of the digestive gland change in response to gonadal energy needs.

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Neuroprotective effect of the water extract of Angelicae Gigantis Radix Palva in ischemic stroke rats (허혈성 뇌졸중 흰쥐에서 당귀미 열수추출물의 혈관신생인자 발현 조절을 통한 신경세포 보호작용에 대한 연구)

  • Park, Ki-Ho;Oh, Tae-Woo;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.27 no.2
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    • pp.61-67
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    • 2012
  • Objective : Angelica Gigas Nakai is a popular oriental medicine used for the treatment of vascular diseases. The aim of this study is to investigate neuroprotective effect of the water extract of Anelicae Gigantis Radix Palva (AG) in transient middle cerebral artery occlusion (tMCAO)-induced ischemic rats via the regulation of angiogenesis-related molecules. Methods : Sprague-Dawley rats were intraperitoneally administrated with AG water extract at doses of 10, 25, 50 mg/kg body weight after tMCAO (90 min occlusion). reperfusion for 24 hr infarction volumes were measured by 2,3,5-tetrazolium chloride (TTC) staining. Brain tissues were observed neuronal cell injuries by nissl staining, and also brain-blood barrier (BBB) permeability change by evans blue. Expression of vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1) and Tie-2 receptor protein in brain tissues was determined by western blot. Results : AG water extract significantly reduced infarction volume in ischemic brains of rats, degradation of neuronal cell, BBB permeability and expression of VEGF protein dose-dependently. Ang-1 protein was increased dose-dependantly, not significantly. Conclusion : This study suggests that AG water extract shows neuroprotective effect by preventing BBB breakdown, with regulating angiogenesis factor VEGF and Ang-1.

Clinical Significance of SH2B1 Adaptor Protein Expression in Non-small Cell Lung Cancer

  • Zhang, Hang;Duan, Chao-Jun;Chen, Wei;Wang, Shao-Qiang;Zhang, Sheng-Kang;Dong, Shuo;Cheng, Yuan-Da;Zhang, Chun-Fang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2355-2362
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    • 2012
  • The SH2B1 adaptor protein is recruited to multiple ligand-activated receptor tyrosine kinases that play important role in the physiologic and pathologic features of many cancers. The purpose of this study was to assess SH2B1 expression and to explore its contribution to the non-small cell lung cancer (NSCLC). Methods: SH2B1 expression in 114 primary NSCLC tissue specimens was analyzed by immunohistochemistry and correlated with clinicopathological parameters and patients' outcome. Additionally, 15 paired NSCLC background tissues, 5 NSCLC cell lines and a normal HBE cell line were evaluated for SH2B1 expression by RT-PCR and immunoblotting, immunofluorescence being applied for the cell lines. Results: SH2B1 was found to be overexpressed in NSCLC tissues and NSCLC cell lines. More importantly, high SH2B1 expression was significantly associated with tumor grade, tumor size, clinical stage, lymph node metastasis, and recurrence respectively. Survival analysis demonstrated that patients with high SH2B1 expression had both poorer disease-free survival and overall survival than other patients. Multivariate Cox regression analysis revealed that SH2B1 overexpression was an independent prognostic factor for patients with NSCLC. Conclusions: Our findings suggest that the SH2B1 protein may contribute to the malignant progression of NSCLC and could offer a novel prognostic indicator for patients with NSCLC.

KLK6 Promotes Growth, Migration, and Invasion of Gastric Cancer Cells

  • Zhu, Shengxing;Shi, Jihua;Zhang, Shanfeng;Li, Zhen
    • Journal of Gastric Cancer
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    • v.18 no.4
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    • pp.356-367
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    • 2018
  • Purpose: Kallikrein (KLK) proteases are hormone-like signaling molecules with critical functions in different cancers. This study investigated the expression of KLK6 in gastric cancer and its potential role in the growth, migration, and invasion of gastric cancer cells. Materials and Methods: In this study, we compared protein levels of KLK6, vascular endothelial growth factor (VEGF), and matrix metallopeptidase (MMP) 9 in normal gastric epithelial and gastric cancer cell lines by western blot. Fluorescence-activated cell sorting was employed to sort 2 clones of SGC-7901 cells with distinct KLK6 expression, namely, KLK6-high ($KLK6^{high}$) and KLK6-low ($KLK6^{low}$), which were then expanded. Lastly, immunohistochemical analysis was performed to investigate KLK6 expression in gastric cancer patients. Results: The expression levels of KLK6, VEGF, and MMP 9, were significantly higher in the gastric cancer cell lines SGC-7901, BGC-823, MKN-28, and MGC-803 than in the normal gastric epithelial cell line GES-1. Compared to $KLK6^{low}$ cells, $KLK6^{high}$ cells showed enhanced viability, colony-forming ability, migration, and invasion potential in vitro. Importantly, immunohistochemical analysis of a human gastric cancer tissue cohort revealed that the staining for KLK6, VEGF, and MMP9 was markedly stronger in the cancerous tissues than in the adjacent normal tissues. KLK6 expression also correlated with that of VEGF and MMP9 expression, as well as several key clinicopathological parameters. Conclusions: Together, these results suggest an important role for KLK6 in human gastric cancer progression.

Various expression patterns of pregnancy-associated plasma protein-A

  • Jeon, Eunjeong;Lee, Jihwan;Son, Junkyu;Kim, Doosan;Lim, Dajeong;Han, Man-Hye;Hwang, Seongsoo
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.3
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    • pp.155-161
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    • 2022
  • Pregnancy-associated plasma protein-A (PAPP-A) is known as an important biomarker for fetal abnormality during first trimester and has a pivotal role in follicle development and corpus luteum formation. And also, it is being revealed that an expression of PAPP-A in various cells and tissues such as cancer and lesion area. PAPP-A is the major IGF binding protein-4 (IGFBP-4) protease. Cleavage of IGFBP-4 results in loss of binding affinity for IGF, causing increased IGF bioavailability for proliferation, survival, and migration. Additionally, PAPP-A can be used as a promising therapeutic target for healthy longevity. Despite growing interest, almost nothing is known about how PAPP-A expression is regulated in any tissue. This review will focus on what is currently known about the zinc metalloproteinase, PAPP-A, and its role in cells and tissues. PAPP-A is expressed in proliferating cells such as fetus in uterus, granulosa cells in follicle, dermis in wound, cancer cells, and Sertoli cells in testis. They have common characteristics of proliferation faster than normal cells with stimulating IGFs action and inhibiting IGFBPs. The PAPP-A functions and expression studies in livestock have not yet been conducted much. Further studies are needed to use PAPP-A as a marker for healthy longevity in animal science.

Interaction between host cell proteins and open reading frames of porcine circovirus type 2

  • Si-Won Park;In-Byung Park;Seok-Jin Kang;Joonbeom Bae;Taehoon Chun
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.698-719
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    • 2023
  • Postweaning multisystemic wasting syndrome (PMWS) is caused by a systemic inflammation after porcine circovirus type 2 (PCV2) infection. It was one of the most economically important pathogens affecting pig production worldwide before PCV2 vaccine was first introduced in 2006. After the development of a vaccine against PCV2a type, pig farms gradually restored enormous economic losses from PMWS. However, vaccine against PCV2a type could not be fully effective against several different PCV2 genotypes (PCV2b - PCV2h). In addition, PCV2a vaccine itself could generate antigenic drift of PCV2 capsid. Therefore, PCV2 infection still threats pig industry worldwide. PCV2 infection was initially found in local tissues including reproductive, respiratory, and digestive tracks. However, PCV2 infection often leads to a systemic inflammation which can cause severe immunosuppression by depleting peripheral lymphocytes in secondary lymphoid tissues. Subsequently, a secondary infection with other microorganisms can cause PMWS. Eleven putative open reading frames (ORFs) have been predicted to encode PCV2 genome. Among them, gene products of six ORFs from ORF1 to ORF6 have been identified and characterized to estimate its functional role during PCV2 infection. Acquiring knowledge about the specific interaction between each PCV2 ORF protein and host protein might be a key to develop preventive or therapeutic tools to control PCV2 infection. In this article, we reviewed current understanding of how each ORF of PCV2 manipulates host cell signaling related to immune suppression caused by PCV2.

Identification of Phospholipase C Activated by $GTP{\gamma}S$ in Plasma Membrane of Oat Cell

  • Kim, Hyae-Kyeong;Park, Moon-Hwan;Chae, Quae
    • BMB Reports
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    • v.28 no.5
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    • pp.387-391
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    • 1995
  • In order to investigate whether phospholipase C (PLC) activity in oat celIs is regulated by Gprotein, we have characterized PLC in plasma membranes of oat tissues. To identify the purified plasma membrane, $K^+$-stimulated, $Mg^{2+}$-dependent ATPase activity was measured. The activity of ATPase was shown to be proportional to the concentration of membrane protein. To examine the PLC activity regulated by G-protein, we used the inside-out and outside-out plasma membrane mixture isolated from the oat cells. The plasma membrane mixture showed higher PLC activity than the one of the outside-out plasma membrane. This suggests that PLC activity is located at the cytoplasmic surface of plasma membrane. PLC activity in plasma membrane mixture was dependent on $Ca^{2+}$ with maximum activity at 100 ${\mu}m$ $Ca^{2+}$ and it was inhibited by 1 mM EGTA. Using Sep-pak $Accell^{TM}$ Plus QMA chromatography, we found that inositol 1,4,5-trisphosphate ($IP_3$) was produced in the presence of 10 ${\mu}m$ $Ca^{2+}$. The PLC activity in the membrane was enhanced by an activator of G-protein ($GTP{\gamma}S$) and not by an inhibitor ($GDP{\beta}S$). This indicates that a G-protein is involved in the activation of PLC in the plasma membrane of oat cells.

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A Study on Bioaccumulation of Heavy Metals in Mussels (Mytilus edulis) from the Onsan Coastal Zone (온산 연안에 서식하는 진주담치(Mytilus edulis)의 중금속 생물농축에 관한 연구)

  • 백수민;이인숙
    • The Korean Journal of Ecology
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    • v.21 no.3
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    • pp.217-224
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    • 1998
  • The heavy metal concentrations of seawater collected from the Onsan coastal zone in February and July 1996 and mussels(Mytilus edulis) in February 1997 were analysed. The concentrations of cadmium in seawater were in the range of 0.008-2.988 ${\mu}g/L$, while the ranges of copper and zinc concentrations were 0.08-2.55, and 0.21-35.12 ${\mu}g/L$, respectively. The metal concentrations decreased gradually with increasing distances from Daejeong stream, indicating that this stream was the major source of heavy metal input into the Onsan coastal zone. The concentrations of cadmium, copper and zinc in mussels were in the ranges of 1.40-25.09, 8.5-64.5, and 46.8-291.2 ${\mu}g/g$, respectively. The metal concentrations decreased gradually with increasing distances from Daejeong stream. Among organs of mussels, gill showed the highest concentrations of cadmium and the digestive gland showed the highest concentrations of copper and for zine the kidney showed the highest concentrations. The digestive gland and kidney revealed high proportion of cadmium in cytosolic fraction and the percentage of copper was high in the kidney and that of zine was high in the digestive gland. Metal-binding protein of mussels collected from the mouth of Daejeong stream was separated, using gel-filtration chromatography. In the kidney and gill of mussels, most of cadmium was associated with metal-binding protein. In contrast, most of the metal in the digestive gland and remaining tissues is bound to high molecular weight protein rather than metal-binding protein.

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