• BMB Reports
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• 제38권3호
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• pp.354-359
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• 2005

Open reading frame 29 (ha29) is a gene specific for Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearSNPV). Sequence analyses showed that the transcription factor Tfb2 motif, bromodomain and Half-A-TPR (HAT) repeat were present at aa 66-82, 4-76, 55-90 of the Ha29 protein respectively. The product of Ha29 was detected in HearSNPV-infected HzAM1 cells at 3 h post-infection. Western blot analysis using a polyclonal antibody produced by immunizing a rabbit with purified GST-Ha29 fusion protein indicates that Ha29 is an early gene. The size of Ha29 product in infected HzAM1 cells was about 25 kDa, which was larger than the presumed size of 20.4 kDa. Tunicamycin treatment of HearSNPV-infected HzAM1 cells suggested that the Ha29 protein is N-glycosylated. Fluorescent confocal laser scanning microscope examination, and Western blot analysis of purified budded virus (BVs), occlusion-derived virus (ODVs), cell nuclear and cytoplasmic fraction, showed that the Ha29 protein was localized in the nucleus. Our results suggested that ha29 of HearSNPV encodes a non-structurally functional protein that may be associated with virus gene transcription in Helicoverpa hosts.

• Biomolecules & Therapeutics
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• 제23권1호
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• pp.19-25
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• 2015

Vascular endothelial growth factor (VEGF) is an important regulator of neovascularization. Hypoxia inducible nitric oxide (NO) enhanced the expression of VEGF and thymosin beta-4 ($T{\beta}4$), actin sequestering protein. Here, we investigated whether NO-mediated VEGF expression could be regulated by $T{\beta}4$ expression in HeLa cervical cancer cells. Hypoxia inducible NO production and VEGF expression were reduced by small interference (si) RNA of $T{\beta}4$. Hypoxia response element (HRE)-luciferase activity and VEGF expression were increased by the treatment with N-(${\beta}$-D-Glucopyranosyl)-N2-acetyl-S-nitroso-D, L-penicillaminamide (SNAP-1), to generate NO, which was inhibited by the inhibition of $T{\beta}4$ expression with $T{\beta}4$-siRNA. In hypoxic condition, HRE-luciferase activity and VEGF expression were inhibited by the treatment with $N^G$-monomethyl-L-arginine (L-NMMA), an inhibitor to nitric oxide synthase (NOS), which is accompanied with a decrease in $T{\beta}4$ expression. VEGF expression inhibited by L-NMMA treatment was restored by the transfection with pCMV-$T{\beta}4$ plasmids for $T{\beta}4$ overexpression. Taken together, these results suggest that $T{\beta}4$ could be a regulator for the expression of VEGF via the maintenance of NOS activity.

• 대한수의학회지
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• 제38권3호
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• pp.581-588
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• 1998

Eight monoclonal antibodies(MAbs) against bovine coronavirus(BCV) were produced and characterized. Three MAbs(1G9, 4H12, 5C1) specific to the S glycoprotein and two HE glycoprotein-specific MAbs(2A5, 5G4) were found to neutralize the BCV in fluorescence focus neutralization(FFN) test. Two HE-specific MAbs from the neutralizing MAbs inhibited the hemagglutinating activity of the BCV. None of the N protein-specific MAbs(1C1, 5A12, 6H1) neutralized the virus infectivity. Bovine coronavirus and mouse hepatitis virus, which belong to group II coronaviruses, were differentiated from other groups of coronaviruses(porcine transmissible gastroenteritis virus, porcine epidemic diarrhea virus, canine coronavirus) by all MAbs in fluorescence antibody test(FA), but not in FFN test.

• 한국가금학회지
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• 제21권4호
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• pp.257-266
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• 1994

Three protein feeding systems for egg-type pullets involving conventional step-down protein 18-15-12%), step-up protein(12-15-18%) and single-stage low protein (13-13-13%) with an iso-energy level of 2,900 ME kcal /kg were compared to examine the effect on pullet growth and subsequent laying performance. During the growing period, pullets subjected to the step-up and single-stage low protein feeding systems were lighter in body weight and consumed less feed and netabolizable energy than those on the conventional step-down protein feeding system(P<0.05). 3ut the pullets on the step-up protein diet consumed more protein, and those on the single-stage low protein diet consumed less protein than those on the step-down protein diet(P<0.05). Also, he feed cost was less in pullets on the single-stage low protein diet than in those on the other systems(P<0.05). During the laying period, sexual maturity was later in hens reared on the step-up and single-stage low protein diets than in those on the step-down protein diet(P<0.05), however, average hen-day egg production and egg weight were not significantly affected by the protein feeding systems in the growing period. Daily feed intake and feed required per egg were significantly reduced in hens on the single-stage low protein diet compared to those on conventional protein feeding system(P<0.05). It was concluded that the 13% single-stage low protein feeding system produced smaller pullets with less feed, energy, protein, and feed cost during the growing period, and hens reared on that system consumed less feed during the laying period without any impairment of production compared to the conventional rearing system.

• BMB Reports
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• 제41권4호
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• pp.294-299
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• 2008

Elevated expression of SMYD3 is a frequent genetic abnormality in several malignancies. Few studies knocking down SMYD3 expression in cervical carcinoma cells have been performed to date. In this paper, we established an inducible short hairpin RNA expression system to examine its role in maintaining the malignant phenotype of HeLa cells. After being induced by doxycycline, SMYD3 mRNA and protein expression were both reduced, and significant reductions in cell proliferation, colony formation and migration/invasion activity were observed in the SMYD3-silenced HeLa cells. The percentage of cells in sub-G1 was elevated and DNA ladder formation could be detected, indicating potent induction of apoptosis by SMYD3 knockdown. These findings imply that SMYD3 plays crucial roles in HeLa cell proliferation and migration/invasion, and that it may be a useful therapeutic target in human cervical carcinomas.

• Parasites, Hosts and Diseases
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• 제52권4호
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• pp.439-441
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• 2014

Toxoplasma gondii is the causative agent of toxoplasmosis with symptoms of congenital neurological and ocular diseases and acquired lymphadenitis, retinochoroiditis, and meningoencephalitis. Small molecules which block the activity of protein kinases were tested in in vitro culture of T. gondii to find new therapeutic drugs of safer and more effective than the combined administration of pyrimethamine and sulfadoxine that sometimes provoke lethal Stevens-Johnson syndrome. Among them, Gefitinib and Crizotinib inhibited intracellular growth of T. gondii in HeLa cells by counting the number of T. gondii per parasitophorous vacuolar membrane whereas Sunitinib did not. Gefitinib inhibited the growth of T. gondii in a dose-dependent manner over $5{\mu}M$ up to the tolerable concentration of HeLa cells and halted the division of the parasite immediately from the time point of treatment. Gefitinib inhibition suggests that tyrosine kinases of EGFR family or other homologous kinases of the parasite itself may be the target to cause the block of T. gondii growth.

• Journal of Nutrition and Health
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• 제14권4호
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• pp.151-161
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• 1981

The objective of this study was to compare the electrolytes contents in skeletal muscles of two different groups of rats, one for control fed 20% casein diet, and the other experimental group recovered from 7% casein diet as protein restriction feeding for two weeks at the beginning of this experiment. Two different comparison criteria were used in this study, one for comparison between animals at the same age groups and the other between groups weighing same body weight as the control groups. Food consumption of animals for experimental and control groups were ad libitum. Following measurement and samples were collected: body weight, five different muscles-anterior tibialis (A.T.) extensor digitorium longus (E.D.L.) soleus, plantaris, gastrocnemius-for electrolytes and protein analysis. The experimental groups showed lower body weight than that of control group. Same trend was showen in protein content in five different muscles. Magnesium and potassium content in four muscles except gastrocnemius showed lower and sodium higher in experimental groups than those in control group at fourteenth day, after recovery from protein restriction, electrolytes content change showed differently in four muscles. Magnesium and potassium contents were increased as protein content in diet. Sodium content decreased as one week intake of recovery diet started, thereafter it was rather slowly increased. Phosphorus content in gastrocnemius at the second week in experimental group was slightly lower than that of control group, and it recovered rapider while at the same body weight group it was higher in experimental group.

• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.372-379
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• 2017

The transport of lysosomal enzymes into the lysosomes depends on the phosphorylation of their chains and the binding of the phosphorylated residues to mannose-6-phosphate receptors. The efficiency of separation depends more on the phosphodiesterases (PDEs) than on the activity of the phosphorylation of mannose residues and can be determined in vitro. PDEs play important roles in regulation of the activation of lysosomes. The expression of proteins was confirmed by western blotting. All PDE4 series protein expression was reduced in high concentrations of rolipram. As a result of observing the fluorescence intensity after rolipram treatment, the lysosomal enzyme was activated at low concentrations and suppressed at high concentrations. High concentrations of rolipram recovered the original function. Antimicrobial activity was not shown in either 10 or $100{\mu}M$ concentrations of rolipram in treated HeLa cells in vitro. However, the higher anticancer activity at lower rolipram concentration was shown in lysosomal enzyme treated with $10{\mu}M$ of rolipram. The anticancer activity was confirmed through cathepsin B and D assay. Tranfection allowed examination of the relationship between PDE4 and lysosomal activity in more detail. Protein expression was confirmed to be reduced. Fluorescence intensity showed decreased activity of lysosomes and ROS in cells transfected with the antisense sequences of PDE4 A, B, C, and D. PDE4A showed anticancer activity, whereas lysosome from cells transfected with the antisense sequences of PDE4 B, C, and D had decreased anticancer activity. These results showed the PDE4 A, B, C, and D are conjunctly related with lysosomal activity.

• Biomolecules & Therapeutics
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• 제4권1호
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• pp.46-50
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• 1996

Inhibition of the growth of human cancer cells by proteins secreted from 373-L1 cells was investigated in the present study. The growth of human cancer cells was inhibited by co-culture with 373-L1 cells under 10% FBS and DME, DME, GIT and serumless medium, respectively. The conditioned medium of cultured 373-L1 cells under serumless medium was concentrated 100-fold through an ultrafiltration cell with a 10,000 molecular weight cutoff at 4$^{\circ}C$ under positive pressure using nitrogen(373-L1 EM). 373-L1 EM inhibited the growth of HeLa, Hep G 2, KHOS-Np, A43l and MCF-7 cells. 3T3-L1 EM was purified with FPLC, DEAE-ion exchange chromatography and phenyl-sepharose chromatography. The major protein of 373-L1 EM has a molecular weight of 66,000-68,000 in SDS-PAGE analysis. The results suggest that the inhibitory activity of 373-L1 EM appears to be due to some protein(m.w.66,000-68,000) secreted by 373-L1 cells.

• Asian-Australasian Journal of Animal Sciences
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• 제19권11호
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• pp.1623-1631
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• 2006

The objective of this study was to evaluate the effect of protein and energy on goats fed oil palm fronds (OPF) as roughages. Twenty-four male Saanen goats aged between 7 and 8 months and weighing $23.4{\pm}1.6kg$ were used in a $2{\times}3$ factorial design. Factors were three levels of urea (3%, 4% or 5%) and two levels of energy (low energy (LE) or high energy (HE)). On average, all parameters measured, including dry matter intake (DMI), nutrient digestibility, digestible nutrient intakes, ruminal ammonia-N ($NH_3$-N), ruminal total volatile fatty acid (total VFA) and individual VFA concentrations (mM/L), microbial N supply, P/E ratio and N retention were higher for HE compared to LE diets. Significant (p<0.05) interactions were found between levels of urea and energy for nonstructural carbohydrate (NSC) and energy (DE) digestibilities, ruminal $NH_3$-N and total VFA concentrations. HE diets had higher N absorption and retention than LE diets. Interactions between urea and energy for plasma urea nitrogen (PUN), heat production (HP), and urine and faeces N excretion were significantly lower (p<0.05) for the HE diets than those recorded for the LE diets. The results indicated that supplementation of energy enhanced utilization of urea and resulted in higher animal performance as a consequence of improved ruminal fermentation, microbial yield and N balance. However, the optimal level of urea supplementation remained at 3% in the HE diet.