Expression of the Na⁺-HCO₃⁻cotransporter and its role in pH_iregulation in guinea pig salivary glands

Li, Jingchao,Koo, Na-Youn,Cho, Ik-Hyun,Kwon, Tae-Hwan,Choi, Se-Young,Lee, Sung J.,Oh, Seog B.,Kim, Joong-Soo,Park, Kyungpyo American Physiological Society 2006 American journal of physiology, Gastrointestinal a Vol.291 No.6

<P>Patterns of salivary HCO3<SUP>−</SUP>secretion vary and depend on species and gland types. However, the identities of the transporters involved in HCO3<SUP>−</SUP>transport and the underlying mechanism of intracellular pH (pHi) regulation in salivary glands still remain unclear. In this study, we examined the expression of the Na<SUP>+</SUP>-HCO3<SUP>−</SUP>cotransporter (NBC) and its role in pHiregulation in guinea pig salivary glands, which can serve as an experimental model to study HCO3<SUP>−</SUP>transport in human salivary glands. RT-PCR, immunohistochemistry, and pHimeasurements from BCECF-AM-loaded cells were performed. The amiloride-sensitive Na<SUP>+</SUP>/H<SUP>+</SUP>exchanger (NHE) played a putative role in pHiregulation in salivary acinar cells and also appeared to be involved in regulation in salivary ducts. In addition to NHE, NBC also played a role in pHiregulation in both acini and ducts. In the parotid gland, NBC1 was functionally expressed in the basolateral membrane (BLM) of acinar cells and the luminal membrane (LM) of ducts. In the submandibular gland, NBC1 was expressed only in the BLM of ducts. NBC1 expressed in these two types of salivary glands takes up HCO3<SUP>−</SUP>and is involved in pHiregulation. Although NBC3 immunoreactivity was also detected in submandibular gland acinar cells and in the ducts of both glands, it is unlikely that NBC3 plays any role in pHiregulation. We conclude that NBC1 is functionally expressed and plays a role in pHiregulation in guinea pig salivary glands but that its localization and role are different depending on the type of salivary glands.</P>

Larvicidal and repellent activities of 33 plant extracts against two mosquitoes as Culex pipiens and Aedes albopictus

Dae-Hyun Yoo,Jun-Won Park,Hyun Kyung Kim,Hyun-Na Koo,Gil-Hah Kim 한국응용곤충학회 2014 한국응용곤충학회 학술대회논문집 Vol.2014 No.04

The larvicidal and repellent activities of 33 plant extracts against two mosquitoes as Culex pipiens and Aedes albopictus were examined using direct contact application for larva and a patch test for adult. Chrysanthemum zawadskii var. alpinum, Cnidium officinale, Ginkgo biloba, Magnolia kobus, and Magnolia denudate at 1,000 ppm caused 100% mortality to Cx. pipiens larva within 24 hr. Ailanthus altissima, Dryopteris crassirhizoma, Houttuynia cordata, Mentha arvensis, Phyllostachys nigra, and Parthenocissus tricuspidata showed over 90% mortality to Cx. pipiens. C. zawadskii var. alpinum, C. officinale, G. biloba, M. kobus, M. denudate, and P. nigra gave 100% mortality at 1,000 ppm to Ae. albopictus. Acorus gramineus, Campanula takesimana, and D. crassirhizoma, showed 97.8%, 94.5%, and 94.4% mortality to Ae. albopictus at 24 hr, respectively. Five plant extracts (C. zawadskii var. alpinum, C. officinale, G. biloba, M. kobus, and M. denudate) showed 100% mortality both Cx. pipiens and Ae. albopictus. However, A. altissima gave 90% larvicidal activity to Cx. pipiens, whereas 7.8% mortality to Ae. albopictus. Several plant extracts which have highly larvicidal activities, were determined repellency against two mosquitoes species.

Insecticidal Activity of Dryopteris crassirhizoma Nakai Against Culex pipiens, Aedes albopictus and Anopheles sinensis

Dae Hyun Yoo,Hyun Kyung Kim,Hyun-Na Koo,Gil-Hah Kim 한국응용곤충학회 2014 한국응용곤충학회 학술대회논문집 Vol.2014 No.10

The insecticidal activities of materialsderived from Dryopteris crassirhizoma Nakai against third instar larvae of three species mosquitoes (Culex pipiens, Aedes albopictus and Anopheles sinnensis) were evaluated using a direct contact mortality bioassay. The methanol extracts of D. crassirhizoma showed 100%, 87.8% and 100% larvicidal activity at 1,000 ppm against Cx. pipiens, Ae. albopictus and An. sinensis, respectively. Hexane fraction showed 100% larvicidal activity three species mosquitoes at 500 ppm after 24 hrs. Purification of the biologically active constituents from the hexane extraction with larvicidal activity was done using silica gel column chromatography. H1 and H3 fractions gave 100% mortality to Cx. pipiens, Ae. albopictus and An. sinensis at 100 ppm. H1 fraction separated with methanol to give a H111 through centrifugation. Fraction of the biologically active constituents from the H3 fraction with larvicidal activity was done using methanol. H31 fraction was determined 100% mortality to Cx. Pipiens, Ae. albopictus and An. sinensis at 50 ppm, respectively. Two fractions were analyzed C14H22O (H111, MW206.0) and C11H14O4 (H31, MW210.08) by GC and GC-MS. D. crassirhizoma derived compounds described herein could be useful for managing filed populations as larvicide of Cx. pipiens, Ae. albopictus and An. sinensis.

Insecticidal activity of Curcuma longa L. against Culex pipiens and Aedes albopictus

Jun-Won Prak,Dae-Hyun Yoo,Hyun Kyung Kim,Hyun-Na Koo,Gil-Hah Kim 한국응용곤충학회 2014 한국응용곤충학회 학술대회논문집 Vol.2014 No.04

The insecticidal activities of materials derived from the rhizomes of turmeric, Curcuma longa L., against third instar larva of Culex pipiens and Aedes albopictus were evaluated using a direct contact mortality bioassay. Curcuma longa L. hexane extraction showed 100% larvicidal activity both two mosquitoes species at 1,000 ppm after treated 24 h. Purification of the biologically active constituents from the hexane extraction with larvicidal activity was done using silica gel column chromatography. H1 fraction gave 100% mortality to C. pipiens and A. albopictus at 100 ppm. H12 fraction was determined 100% and 87.8% larvicidal activity to C. pipiens and A. albopictus at 50 ppm, respectively. H12 fraction was analyzed as the sesquiterpene, ar-turmerone (C15H20O) and turmerone(C15H22O) by GC and GC-MS.

Sequence analysis and expression of three major glycoproteins of Aujeszky’s disease virus NYJ strain in Bm5 Cells

Hyun Na Koo,Jeong Mi Oh,Jae Young Choi,Kwang Sik Lee,Jong Yul Roh,Yeon Ho Je,Byung Rae Jin,Sung Sik Yoo,Jae Su Kim,Young In Kim,In Joong Yoon,Soo Dong Woo 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.10

Aujeszky’s disease (AD), also called pseudorabies, is an infectious viral disease, caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. AD affects many countries and regions in the world, causing important economic losses, mainly due to international trade restrictions. In this study, to determine the characteristics of the Aujeszky’s disease virus (ADV), NYJ strain, which was isolated from the serum of an infected pig in 1987, we investigated the nucleotide sequence and expression of the glycoproteins gB, gC, and gD using the bBpGOZA system. We found that the glycoproteins gB, gC, and gD of NYJ consisted of 2751 bp, 1443 bp, and 1203 bp, respectively. Comparison of the NYJ with the other strains revealed nucleotide sequence identity ranging from 91.tito 99.0%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The NYJ strain was formed a distinct branch with high bootstrap support. The expression of glycoprotein gD in insect cells was characterized by SDS-PAGE and Western blotting with an anti-ADV polyclonal antibody. Glycoprotein gD of approximately 45 kDa was detected. The results of this study have implications for both the taxonomy of ADV and vaccine development.

Ginseng radix induces apoptosis in HL-60 cells and its mechanism as little relation with TNF-α production

Koo, Hyun-Na,Shin, Soon-Shik,Park, Jin-Han,Kim, Na-Hyung,Hong, Seung-Heon,Kim, Hyung-Min Kyung Hee Oriental Medicine Research Center 2004 Oriental pharmacy and experimental medicine Vol.4 No.3

Ginseng radix, the root of Panax ginseng C. A. Meyer (Araliaceae), is a medicinal plant used world-widely and has been reported to have various biological effects. To investigate the effects of Ginseng radix on HL-60 cell apoptosis, MTT assay, DNA fragmentation assay and flow cytometry were performed on HL-60 cells. Cells were treated with Ginseng radix at different concentrations $(10^{-4},\;10^{-3}\;and\;10^{-2};\;dilution\;rate)$. Ginseng radix significantly induced cells apoptosis with a time- and dose-dependent manner. To determine whether Ginseng radix-induced apoptosis is due to increase of tumor necrosis factor $(TNF-{\alpha})$ secretion, enzyme-linked immunosorbent assay was performed on HL-60 cells. Unexpectedly, Ginseng radix $(96\;{\pm}\;5\;pg/ml)$ significantly decreased the $TNF-{\alpha}$ secretion compared with control $(174\;{\pm}\;14\;pg/ml)$. Furthermore, Ginseng radix with $rIFN-{\gamma}$ synergistically increased nitric oxide production in mouse peritoneal macrophages. Taken together, our data indicate that Ginseng radix induce apoptosis on HL-60 cells without increase of $TNF-{\alpha}$ secretion and could be used for a supplementary remedy of cancer.

Phenotype and gene expression comparisons between imidacloprid resistant and susceptible strains of cotton aphid by various environmental pressures

Hyun-Na Koo,Hyun Kyung Kim,Gil-Hah Kim 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.04

The melon and cotton aphid Aphis gossypii Glover (Hemiptera; Aphididae) is one of the most serious pests worldwide. We surveyed insecticide susceptibility in A. gossypii field populations to 12 insecticides to examine resistance ratios. The levels of insecticide resistance were extremely high, especially to neonicotinoids. One point mutation was found in the beta1 subunit loop D region of the nicotinic acetylcholine receptor (nAChR) of the imidacloprid-resistant strain. Feeding behavior analysis using an electrical penetration graph showed that sublethal doses of imidacloprid had significant effects on the duration of phloem ingestion. In addition, higher doses of imidacloprid induced contact toxicity rather than inhibition of feeding behavior. Temperature and insecticide are two important factors that affect survival, reproduction and other physiological processes of insects. To determine interactions of temperature and insecticide treatment on susceptible and imidacloprid-resistant strains of A. gossypii, adults were exposed to three temperatures (17, 22, and 28℃) or combinations of three temperatures and imidacloprid (LC20), and the expression of several genes (heat shock protein 70, cuticle protein, cytochrome P450, and elongation factor) were analyzed. Additionally, the effect of electron beam irradiation on development, reproduction, and several gene expression of imidacloprid-resistant and -susceptible A. gossypii were compared.

Enhanced expression of the structural protein of porcine reproductive and respiratory syndrome virus (PRRSV) by SUMO fusion

( Hyun Na Koo ),( Sung Min Bae ),( Soo Dong Woo ) 한국잠사학회 2016 International Journal of Industrial Entomology Vol.32 No.2

The major structural proteins of porcine reproductive and respiratory syndrome virus (PRRSV) are derived from ORFs 4, 5, and 6. They have been considered very important to arouse the humoral and cellular immune responses against PRRSV infection and proposed to be the excellent candidate proteins in the design of PRRS bioengineering vaccine. However, the PRRSV structural proteins are produced in low levels in the infected cells because it forms insoluble protein and possesses several transmembrane regions. To overcome this problem, we fused the ORF4, ORF5, and ORF6 with SUMO (small ubiquitin-related modifier). The resulting fusion protein SUMO-ORF4, -ORF5, and -ORF6 were highly expressed in Bm5 cells. The level of protein expression using the Bombyx mori larvae was higher than that using Bm5 cells. In addition, fusion to SUMOstar, which is not processed by native SUMO proteases, significantly enhanced protein expression levels compared to SUMO fusion. This study demonstrated that SUMO or SUMOstar, when fused with PRRSV structural proteins, was able to promote its soluble expression. This may be a better method to produce PRRSV structural proteins for vaccine development.

High-level expression of the PRRSV structural proteins by SUMO fusion in Bombyx mori cells and larvae

Hyun Na Koo,Sung Min Bae,Tae Young Shin,Jae Bang Choi,Bit Na Rae Yun,Jae Young Choi,Kwang Sik Lee,Jong Yul Roh,Yeon Ho Je,Byung Rae Jin,Soo Dong Woo 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.05

The porcine reproductive and respiratory syndrome virus (PRRSV) has three major structural proteins which designated as GP4, GP5, and M. They have been considered very important to arouse the humoral and cellular immune responses against PRRSV infection and proposed to be the excellent candidate proteins in the design of PRRS bioengineering vaccine. However, the PRRSV structural proteins are produced in low levels in the infected cells because it forms insoluble protein and possesses several transmembrane regions. To overcome this problem, we fused the GP4, GP5, and M with SUMO (Small ubiquitin-related modifier), and expressed the fused gene in Bm5 cells and silkworm larvae. Expression of the proteins were analyzed by 12% SDS-PAGE and western blotting using 6xHis tag and porcine anti-PRRSV antibodies. In results, SUMO fused proteins were expressed at a high level in Bm5 cells. The levels of protein using the silkworm larvae is higher than that using Bm5 cells. The fused protein was purified by Ni-NTA affinity chromatography. This study demonstrated that SUMO, when fused with PRRSV structural proteins, was able to promote its soluble expression. This may be a better method to produce PRRSV structural proteins for vaccine development.

Molecular Characterization of ORFs 2 to 7 of Korean Porcine Reproductive and Respiratory Syndrome Virus (CA) and Its Protein Expression by Recombinant Baculoviruses

Hyun Na Koo,Jeong Mi Oh,Jae Kyung Lee,최재영,이광식,노종열,제연호,진병래,유성식,Jae Su Kim,Young In Kim,윤인중,우수동 한국미생물학회 2008 The journal of microbiology Vol.46 No.6

To determine the characteristics of the Korean porcine reproductive and respiratory syndrome virus (PRRSV), CA, which was isolated from the serum of an infected pig in 2006, we investigated the nucleotide sequence and expression of the structural ORFs (ORFs 2 to 7) using the bApGOZA system. We found that the structural ORFs 2 to 7 of CA consisted of 3188 nucleotides that were the same as those formed from VR-2332. Comparison of the CA with the other strains revealed nucleotide sequence identity ranging from 89.8 to 99.5%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The CA strain was closely related to the other North American genotype strains but formed a distinct branch with high bootstrap support. Additionally, expression levels of the PRRSV proteins in insect cells were strong or partially weak. The results of this study have implications for both the taxonomy of PRRSV and vaccine development.