• Title/Summary/Keyword: Alternative protein

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$p19^{ras}$ Accelerates $p73{\beta}$-mediated Apoptosis through a Caspase-3 Dependent Pathway

  • Jang, Sang-Min;Kim, Jung-Woong;Choi, Kyung-Hee
    • Animal cells and systems
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    • v.13 no.4
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    • pp.399-403
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    • 2009
  • $p19^{ras}$ is an alternative splicing variant of the proto-oncogene c-H-ras pre-mRNA of $p21^{ras}$. In contrast to $p21^{ras}$, $p19^{ras}$ does not have a C-terminal CAAX motif that targets the plasma membrane and is localized to both the cytoplasm and nucleus. We found that $p19^{ras}$ activated the transcriptional activity of $p73{\beta}$ through protein-protein interactions in the nucleus. p73 is known to play an important role in cellular damage responses such as apoptosis. Although p73 is a structural and functional homologue of p53, p73-mediated apoptosis has not yet been clearly elucidated. In this study, we demonstrate that the interaction between $p19^{ras}$ and $p73{\beta}$ accelerated $p73{\beta}$-induced apoptosis through a caspase-3 dependent pathway. Treatment with DEVD-CHO, a caspase inhibitor, also strengthened $p73{\beta}$-mediated apoptosis through a caspase-3 dependent pathway. Furthermore, the enhanced transcriptional activity of endogenous $p73{\beta}$ by treatment with Taxol was amplified by $p19^{ras}$ overexpression, which markedly increased caspase-3 dependent apoptosis in the p53-null SAOS2 cancer cell line. Our findings indicate a functional linkage between $p19^{ras}$ and p73 in caspase-3 mediated apoptosis of cancer cells.

The Effect of Taping Therapy on the Expression of cFos Protein and Pain Suppression in Acute Ankle Sprain in Rats (흰쥐의 급성 발목삠에서 테이핑요법이 cFos 단백의 발현과 통증억제에 미치는 영향)

  • Choi, Suck-Jun;Byun, Sin-Kyu;Lee, Gyoung-Wan;Kim, Jae-Hyo;Yang, Seung-Bum
    • Korean Journal of Acupuncture
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    • v.34 no.1
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    • pp.1-7
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    • 2017
  • Objectives : The purpose of this study was to evaluate effects of taping therapy on recovery of behavioral symptoms and neural excitability of the lumbar spinal cord in rat model for ankle sprain. Methods : Adult Sprague-Dawley rats was used and divided into 3 experimental groups: normal group(n=6), ankle sprain(n=6), and ankle sprain with taping treatment(n=6). In order to induce ankle sprain the right ankle joint was injured with 4~5 repetitive over-flexions and over-extensions manually. The severity of joint pain was evaluated by measuring foot weight bearing force ratio(FWBRF) of the hind limb and the injury-induced edema formation by diameter of the joint following ankle sprain. The changes of neural excitability in the lumbar spinal cord was tested by observation of cFos protein expression, a metabolic marker for neural excitation. Results : Severity of ankle injury induced in this experiment coincided with Grade 1 ankle sprain. Compared with ankle sprain group, ankle sprain+taping showed a significant reductions of joint pain as well as of edema formation at the ankle joint following ankle sprain. There was significant upregulation of cFos-immunoreactive neurons in the lumbar spinal cord 24 hours after ankle sprain. In contrast, taping therapy resulted in significant inhibition of cFos-immunoreactive neurons in the lumbar spinal cord. Conclusions : Collectively, these results suggest that taping therapy may be an alternative therapeutic intervention for symptom recovery of the mild ankle sprain.

Dormancy Breaking by Hot-water Treatment of Lilium Oriental Hybrids Bulblets Regenerated in Vitro (온탕처리에 의한 오리엔탈나리 소인경의 휴면 타파)

  • Woo, Jin Ha;Nam, Hyo Hoon;Lee, Hyun Suk;Choi, Dong Jin;Oh, Wook;Kim, Kiu Weon
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.2
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    • pp.93-97
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    • 2010
  • The objective of this study was to investigate the effect of hot water and dipping times, and hot- PGR's solution on dormancy breaking of Lilium Oriental Hybrid 'Casa Blanca' and 'Siberia' bulblets regenerated in vitro. Hot-water treatments with distilled water were effective not sprouting rate but shortening of days to sprouting. While $100mg{\cdot}L^{-1}\;GA_{4+7}$ was effective in the dormancy breaking process of bulblets. The content of mitochondrial protein and fumarase activity were highest in bulblet of $100mg{\cdot}L^{-1}\;GA_{4+7}$ addition in hot water treatment. Therefore $GA_{4+7}$ addition in hot-water treatment seem to be the alternative dormancy breaking method to low-temperature treatment in Lilium Oriental Hybrid bulblet regenerated in vitro.

RRM but not the Asp/Glu domain of hnRNP C1/C2 is required for splicing regulation of Ron exon 11 pre-mRNA

  • Moon, Heegyum;Jang, Ha Na;Liu, Yongchao;Choi, Namjeong;Oh, Jagyeong;Ha, Jiyeon;Kim, Hyeon Ho;Zheng, Xuexiu;Shen, Haihong
    • BMB Reports
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    • v.52 no.11
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    • pp.641-646
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    • 2019
  • The Ron proto-oncogene is a human receptor for macrophage-stimulating protein (MSP). The exclusion of exon 11 in alternative splicing generates ${\Delta}RON$ protein that is constitutively activated. Heterogenous ribonucleaoprotein (hnRNP) $C_1/C_2$ is one of the most abundant proteins in cells. In this manuscript, we showed that both hnRNP $C_1$ and $C_2$ promoted exon 11 inclusion of Ron pre-mRNA and that hnRNP $C_1$ and hnRNP $C_2$ functioned independently but not cooperatively. Moreover, hnRNP $C_1$ stimulated exon 11 splicing through intron 10 activation but not through intron 11 splicing. Furthermore, we showed that, whereas the RRM domain was required for hnRNP $C_1$ function, the Asp/Glu domain was not. In conclusion, hnRNP $C_1/C_2$ promoted exon 11 splicing independently by stimulating intron 10 splicing through RRM but not through the Asp/Glu domain.

Biosynthesis of trifolin, a bioactive flavonoid by biotransformation (생물전환으로 생리활성물질인 trifolin의 생합성)

  • Noh, Hye-Ryeong;Kang, Ju-Yeong;Kim, Bong-Gyu
    • Journal of Applied Biological Chemistry
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    • v.64 no.3
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    • pp.309-316
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    • 2021
  • Kaempferol 3-O-galactoside (Trifolin), a member of the flavonol group, has been reported to have anticancer effects against promyelocytic leukemia, histocytic lymphoma, skin melanoma and lung cancer. Trifolin has been extracted and used from several plants, but the extraction process is complicated and the final yield is low. Biotransformation is an alternative tool to produce high value-added chemicals from inexpensive compounds. To synthesis trifolin from naringenin, three genes (PeFLS and OsUGE-PhUGT) were introduced into Escherichia coli, respectively. In order to synthesis trifolin from naringenin, a co-culture fermentation system was established by optimizing the cell concentration, biotransformation temperature and medium, isopropyl-β-D-thiogalactoside (IPTG) concentration, substrate supply concentration, and recombinant protein induction time. The established optimal conditions for trifolin production were a 3:1 ratio of BL-UGTE to BL-FLS, induction of recombinant protein at 25 ℃ for 4 h after addition of 2.0 mM IPTG, biotransformation at 30 ℃, and supply of 300 μM naringenin. Through the optimized co-culture fermentation system, trifolin was biosynthesized up to 67.3 mg/L.

Role of Organic Spices in the Preservation of Traditionally Fermented Kunun-zaki

  • Williana, N. Mokoshe;Babasola, A. Osopale;Cajethan, O. Ezeamagu;Fapohunda, Stephen O.
    • Microbiology and Biotechnology Letters
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    • v.49 no.2
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    • pp.192-200
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    • 2021
  • Kunun-zaki, produced by submerged fermentation of a combination of millet and sorghum, is a popular beverage in Northern Nigeria. Owing to the nature of the process involved in its production, kunun-zaki is highly susceptible to contamination by food spoilage microorganisms, leading to inconsistent quality and short shelf-life. In this study, we investigated various food spices, including cinnamon, garlic, and nutmeg, as potential preservatives that could be used to extend kunun-zaki shelf-life. Kunun-zaki varieties were fermented with each of these spices mentioned above and subjected to bacterial, nutritional, sensory, and quality maintenance assessments (using a twelve-member sensory panel to evaluate the organoleptic properties of kunun-zaki). Bacterial counts in the final products ranged between 105-7 CFU/ml. We identified two bacterial genera, Weissella and Enterococcus, based on partial 16S rRNA gene amplicon sequencing. Three amino acids, namely leucine, aspartate, and glutamate, were abundant in all kunun-zaki varieties, while the total essential amino acid content was above 39%, suggesting that kunun-zaki could potentially be considered as a protein-rich food source both for infants and adults. The kunun-zaki products were also rich in carbohydrates, crude proteins, ash, crude fiber, and fat, with contents estimated as 81-84, 8-11, 0.8-4.0, 2.9-3.58, and 5.1-6.3%, respectively. However, this nutritional content depreciated rapidly after 24 h of storage, except for kunun-zaki fermented with garlic, which its crude protein and fat content was maintained for up to 48 h. Our results revealed that organic spices increased the nutritional content of the kunun-zaki varieties and could be potentially be used as natural preservatives for enhancing the kunun-zaki shelf-life. However, garlic might be considered a better alternative based on our preliminary investigation. The presence of the isolated microorganisms in the analyzed kunun-zaki samples should be highlighted to raise awareness on the possible health hazards that could arise from poor handling and processing techniques.

The alternative approach of low temperature-long time cooking on bovine semitendinosus meat quality

  • Ismail, Ishamri;Hwang, Young-Hwa;Bakhsh, Allah;Joo, Seon-Tea
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.2
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    • pp.282-289
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    • 2019
  • Objective: This study aimed to elucidate whether innovative sous vide treatment has a significant influence on the beef semitendinosus muscle as compared to common sous vide treatment and traditional cooking. Methods: The innovative sous vide treatments were cooked at $45^{\circ}C$ and $65^{\circ}C$ for 6 h (SV45-65), common sous vide treatment at $45^{\circ}C$ and $65^{\circ}C$ for 3 h (SV45 and SV65) and traditional cooking at $75^{\circ}C$ for 30 min (CON75). Water loss and cooking loss, as well as the physical properties (color and shear force) and chemical properties (protein and collagen solubility) of the treated meat, were investigated. Results: The results obtained indicated that the innovative sous vide with double thermal treatment (SV45-65) and cooked with air presence (CON75) resulted to lower $a^*$ and higher $b^*$ values, respectively. The water loss and cooking loss increased when temperature increased from $45^{\circ}C$ to $65^{\circ}C$, and lower water loss was recorded in SV45 and CON75. These samples presented higher water content and revealed strong correlation to protein solubility. Warner-Bratzler shear force (SF) analysis showed the marked interaction between cooking temperature and time. Sample cooked at a high temperature (CON75) and a long period (SV45-65) showed a significantly lower value of SF than sample SV65 (p<0.05). Interestingly, there was no difference in SF values between SV45-65 and CON75. Conclusion: The innovative sous vide treatment with double thermal effect appears an attractive cooking method as compared to common sous vide and traditional cooking method, as it has a potential for improving tenderness values of cooked beef semitendinosus muscle.

Effect of fermented spent instant coffee grounds on milk productivity and blood profiles of lactating dairy cows

  • Choi, Yongjun;Rim, Jongsu;Lee, Honggu;Kwon, Hyunchul;Na, Youngjun;Lee, Sangrak
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.1007-1014
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    • 2019
  • Objective: This study was conducted to evaluate the fermentation characteristics under low mesophilic temperature of spent instant coffee ground (SICG) and to estimate the effect of fermented SICG (FSICG) as alternative feed ingredient on milk productivity of dairy cows. Methods: In the fermentation trial, fermentation of SICG was performed to investigate changes in characteristics using the microbial mixture (Lactobacillus plantarum, Saccharomyces cerevisiae, and Bacillus subtilis = 1:1:1) for 21 days at $20^{\circ}C$ under anaerobic conditions. Molasses was added at 5% of dry mass. In the animal trial, eighteen Holstein Friesian cows were used to evaluate the nutritive value of the FSICG which was fermented for 14 days under the same condition as the fermentation trial. Results: In the fermentation trial, the dry matter (DM) and organic matter content linearly decreased with fermentation time (p<0.001 and p = 0.008, respectively). The acid detergent insoluble nitrogen content linearly decreased with fermentation time (p = 0.037). The microorganism counts linearly increased for Lactobacillus plantarum, Saccharomyces cerevisiae, and Bacillus subtilis across fermentation time (p<0.001). In the animal trial, the DM intake of the control and FSICG treatment were not significantly different, as were milk yield, 4% fat corrected milk, fat-protein corrected milk, and feed to milk conversion content. Fat, protein, lactose, non-fat solids, milk urea nitrogen, and somatic cell counts were also not significantly different in milk composition between treatments. Conclusion: FSICG should be considered a sufficient substitute for cottonseed as a feed component, and 5% DM of a dietary FSICG level was appropriate for dairy cow diets.

An Engineered Outer Membrane-Defective Escherichia coli Secreting Protective Antigens against Streptococcus suis via the Twin-Arginine Translocation Pathway as a Vaccine

  • Li, Wenyu;Yin, Fan;Bu, Zixuan;Liu, Yuying;Zhang, Yongqing;Chen, Xiabing;Li, Shaowen;Li, Lu;Zhou, Rui;Huang, Qi
    • Journal of Microbiology and Biotechnology
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    • v.32 no.3
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    • pp.278-286
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    • 2022
  • Live bacterial vector vaccines are one of the most promising vaccine types and have the advantages of low cost, flexibility, and good safety. Meanwhile, protein secretion systems have been reported as useful tools to facilitate the release of heterologous antigen proteins from bacterial vectors. The twin-arginine translocation (Tat) system is an important protein export system that transports fully folded proteins in a signal peptide-dependent manner. In this study, we constructed a live vector vaccine using an engineered commensal Escherichia coli strain in which amiA and amiC genes were deleted, resulting in a leaky outer membrane that allows the release of periplasmic proteins to the extracellular environment. The protective antigen proteins SLY, enolase, and Sbp against Streptococcus suis were targeted to the Tat pathway by fusing a Tat signal peptide. Our results showed that by exploiting the Tat pathway and the outer membrane-defective E. coli strain, the antigen proteins were successfully secreted. The strains secreting the antigen proteins were used to vaccinate mice. After S. suis challenge, the vaccinated group showed significantly higher survival and milder clinical symptoms compared with the vector group. Further analysis showed that the mice in the vaccinated group had lower burdens of bacteria load and slighter pathological changes. Our study reports a novel live bacterial vector vaccine that uses the Tat system and provides a new alternative for developing S. suis vaccine.

Oral administration of Jinan Red Ginseng and licorice extract mixtures ameliorates nonalcoholic steatohepatitis by modulating lipogenesis

  • Yang, Daram;Jeong, Hyuneui;Hwang, Seung-Mi;Kim, Jong-Won;Moon, Hee-Won;Lee, Ye-Eun;Oh, Hyo-Bin;Park, Chung-berm;Kim, Bumseok
    • Journal of Ginseng Research
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    • v.46 no.1
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    • pp.126-137
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    • 2022
  • Background: Nonalcoholic steatohepatitis (NASH) is one of the main chronic liver diseases. NASH is identified by lipid accumulation, inflammation, and fibrosis. Jinan Red Ginseng (JRG) and licorice have been widely used because of their anti-inflammatory and hepatoprotective effects. Hence, this study assessed JRG and licorice extract mixtures' effects on NASH progression. Methods: Palmitic acid (PA) and the western diet (WD) plus, high glucose-fructose water were used to induce in vitro and in vivo NASH. Mice were orally administered with JRG-single (JRG-S) and JRG-mixtures (JRG-M; JRG-S + licorice) at 0, 50, 100, 200 or 400 mg/kg/day once a day during the last half-period of diet feeding. Results: JRG-S and JRG-M reduced NASH-related pathologies in WD-fed mice. JRG-S and JRG-M consistently decreased the mRNA level of genes related with inflammation, fibrosis, and lipid metabolism. The treatment of JRG-S and JRG-M also diminished the SREBP-1c protein levels and the p-AMPK/AMPK ratio. The FAS protein levels were decreased by JRG-M treatment both in vivo and in vitro but not JRG-S. Conclusion: JRG-M effectively reduced lipogenesis by modulating AMPK downstream signaling. Our findings suggest that this mixture can be used as a prophylactic or therapeutic alternative for the remedy of NASH.