• Food Science of Animal Resources
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• v.30 no.4
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• pp.575-581
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• 2010

This investigation was aimed at developing a ready-to-reconstitute beverage by utilizing probiotics and whey protein hydrolysates carrying bioactive peptides. Cheddar cheese whey was ultrafiltered. The 18% protein retentate was subjected to protein hydrolysis using Neutrase. The hydrolyzed retentate was further condensed to 35% total solids and spray-dried at $75^{\circ}C$ outlet air temperature. Different levels of sugar, citric acid and stabilizer were blended for spray-dried hydrolysates. Spray-dried hydrolysate was further inoculated with different levels of probiotics grown in a whey medium and dried in fluidized-bed drier at $40^{\circ}C$ to obtain a ready-to-reconstitute beverage. Hydrolysis was greatest at an enzyme:substrate ratio of 1:25 for 3 h. Spray-dried hydrolysate reconstituted to 1% protein and blended with 15% sugar, 0.2% citric acid and 0.15% xantham gum resulted in a superior product with no sedimentation. Accordingly, sugar, citric acid and xanthum gum were dry-blended with spray-dried hydrolysates. Bifidobacterium bifidum and Lactobacillus acidophilus that was grown separately in a whey medium, blended to produce 2% spray-dried hydrolysate and dried as described above resulted in a readyto-reconstitute beverage mix. The fluidized dried product typically exhibited a probiotic count of $10^8$colony forming units (CFU)/g. However, blending of probiotic to the retentate and direct spray-drying precipitously reduced the probiotic count to $10^4$ CFU/g of powder.

• Journal of Plant Biotechnology
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• v.47 no.4
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• pp.273-282
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• 2020

In Vietnam, Celastrus hindsii Benth, a medicinal plant rich in secondary metabolites, has been used to alleviate distress caused by ulcers, tumors, and inflammation for generations. The occurrence of two phenotypes, Broad Leaf (BL) and Narrow Leaf (NL), has raised questions about the selection of appropriate varieties for conservation and crop improvement to enhance medicinal properties. This study examined molecular differences in C. hindsii by comparing protein profiles between the NL and BL types using 2D-PAGE and MS. Peptide sequences and proteins were identified by matching MS data against the MSPnr100 databases and verified using the MultiIdent tool on ExPASy and the Blast2GO software. Our results revealed notable variations in protein abundance between the NL and BL proteomes. Selected proteins were confidently identified from 12 protein spots, thereby highlighting the molecular variation between NL and BL proteomes. Upregulated proteins in BL were found to be associated with flavonoid and amino acid biosynthesis as well as nuclease metabolism, which probably attributed to the intraspecific variations. Several bioactive proteins identified in this study can have applications in cancer therapeutics. Therefore, the BL phenotype characterized by healthier external morphological features has higher levels of bioactive compounds and could be better suited for medicinal use.

• Molecules and Cells
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• v.26 no.5
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• pp.496-502
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• 2008

Cryparin, encoded as a single copy gene (Crp) of the chestnut blight fungus Cryphonectria parasitica, is the most abundant protein produced by this fungus. However, its accumulation is decreased remarkably in C. parastica strains containing the double-stranded (ds) RNA virus Cryphonectria hypovirus 1. To characterize the transcriptional regulatory element(s) for strong expression and viral regulation, promoter analysis was conducted. Serial deletion of the Crp promoter region resulted in a step-wise decrease in promoter activity, indicating a localized distribution of genetic elements in the cryparin promoter. Promoter analysis indicated two positive and a repressive cis-acting elements. Among them, the promoter region between nt -1,282 and -907 appeared to be necessary for hypoviral-mediated down-regulation. An electrophoretic mobility shift assay (EMSA) on the corresponding promoter region (-1,282/-907) indicated two regions at (-1,257/-1,158) and (-1,107/-1,008) with the characteristic AGGAGGA-N42-GAGAGGA and its inverted repeat TCCTCTC-N54-TCCTCCT, respectively, appeared to be specific binding sites for cellular factors.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.7
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• pp.3293-3297
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• 2014

Background: Pine needle oil from crude extract of pine needles has anti-tumor effects, but the effective component is not known. Methods: In the present study, compounds from a steam distillation extract of pine needles were isolated and characterized. Alpha-pinene was identified as an active anti-proliferative compound on hepatoma carcinoma BEL-7402 cells using the MTT assay. Results: Further experiments showed that ${\alpha}$-pinene inhibited BEL-7402 cells by arresting cell growth in the G2/M phase of the cell cycle, downregulating Cdc25C mRNA and protein expression, and reducing cycle dependence on kinase 1(CDK1) activity. Conclusion: Taken together, these findings indicate that ${\alpha}$-pinene may be useful as a potential anti-tumor drug.

• Journal of Plant Biotechnology
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• v.7 no.2
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• pp.97-103
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• 2005

This study was carried out to obtain basic information for gene manipulation in potent edible tobacco (Nicotiana tabacum cv. TI 516). N. tabacum cv. TI 516 is a plant for a possible candidate to use as an edible vaccine, since it contains a low level of nicotine. The effective plant regeneration system through leaf disc culture was achieved using a MS basal medium supplemented with 0.1 mg $1^{-1}$ NAA and 0.5 mg $1^{-1}$ BA. In order to transform the N. tabacum cv. TI 516 with the green fluorescent protein (GFP) gene, Agrobacterium tumefaciens LBA 4404 containing the GFP gene was used. Genomic PCR confirmed the integration of the GFP gene into nuclear genome of transgenic plants. Expression of the GFP gene was identified in callus, apical meristem and root tissue of transgenic N. tabacum cv. TI 516 plants using fluorescence microscopy. Western blot analysis revealed the expression of GFP protein in the transgenic edible tobacco plants. The amount of GFP protein detected in the transgenic tobacco plants was approximately 0.16% of the total soluble plant protein (TSP), which was determined by ELISA.

• Animal Bioscience
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• v.35 no.2_spc
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• pp.317-331
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• 2022

The aim of the present investigation is to determine the nutritional composition of various insects and their potential uses as alternative protein sources in animal diets. The feeding industry requires production systems that use accessible resources, such as feed resources, and concentrates on the potential impacts on production yield and nutritional quality. Invertebrate insects, such as black soldier flies, grasshoppers, mealworms, housefly larvae, and crickets, have been used as human food and as feed for nonruminants and aqua culture while for ruminants their use has been limited. Insects can be mass-produced, participating in a circular economy that minimizes or eliminates food- and feed-waste through bioconversion. Although the model for formula-scale production of insects as feed for domestic animals has been explored for a number of years, significant production and transformation to being a conventional protein resource remains to be deeply investigated. This review will focus on the nutritional composition of various insects and their potential use as alternative protein sources, as well as their potential use to promote and support sustainable animal production. Furthermore, nutritional compositions, such as high protein, lauric acid omega 6, and omega 3, and bioactive compounds, such as chitin, are of great potential use for animal feeding.

• Journal of Life Science
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• v.33 no.1
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• pp.64-72
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• 2023

Despite several advances in identification of cardiac transcription factors, there are still needs to find new bioactive molecules that promote cardiomyogenesis from stem cells to highly efficient myocardial differentiation. We analyzed Illumina expression microarray data of mouse embryonic stem cells (mESCs)-derived cardiomyocytes. 276 genes were upregulated (≥ 4fold) in mESCs-derived cardiomyocytes compared undifferentiated ESCs. Secreted phosphoprotein 2 (Spp2) is one of candidates and is known to inhibit bone morphogenetic protein 2 (BMP2) signal transduction as a pseudoreceptor for BMP2. However, its function in cardiomyogenesis is unknown. We confirmed that Spp2 expression increased during the differentiation into functional cardiomyocytes using mESCs, TC-1/Kh2 and E14. Interestingly, Spp2 secretion transiently increased 3 days after formation of embryoid bodies (EBs), indicating that the extracellular secretion of Spp2 is involved in the differentiation of ESCs into cardiomyocytes. To characterize Spp2, we performed experiments using the C2C12 mouse myoblast cell line, which has the property of shifting the differentiation pathway from myoblastic to osteoblastic by treatment with BMP2. Similar to the differentiation of ESCs, transcription of Spp2 increased as C2C12 myoblasts differentiated into myotubes. In particular, Spp2 secretion increased dramatically in the early stage of differentiation. Furthermore, treatment with Spp2-Flag recombinant protein promoted the differentiation of C2C12 myoblasts into myotubes. Taken together, we suggest a novel bioactive protein Spp2 that differentiates ESCs into cardiomyocytes. This may be useful for understanding the molecular pathways of cardiomyogenesis and for experimental or clinical promotion of stem cell therapy for ischemic heart diseases.

• Journal of Life Science
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• v.28 no.2
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• pp.229-239
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• 2018

As customers pay more attention to choosing food that will support their health, many people in the academic and industrial world have focused on developing foods made with bioactive components. Thus, the use of bioactive components rather than synthetic materials has increased. Because there are no limits to how bioactive components can be used, customers assume they are highly reliable and healthy to consume. In the present study, imitation crab stick samples were made from Alaska Pollack with breast recovered protein from spent laying hens and silkworm cocoon powder (10 g) (T1), Alaska Pollack with breast recovered protein from spent laying hens and silkworm cocoon powder (5 g) + cordyceps powder (5 g) (T2), and Alaska Pollack with breast recovered protein from spent laying hens and cordyceps powder (5 g) + conjugated linoleic acid (CLA) (5 g) (T3). The pH and shear force increased after 2 weeks of storage in all three samples. Shear force was significantly higher in the T3 sample in comparison to the T1 and T2 samples. In meat color, redness ($a^{\ast}$) and whiteness (W) increased as the storage periods increased in all three samples, whereas yellowness ($b^{\ast}$) decreased during storage. The T2 sample was significantly higher in redness ($a^{\ast}$), yellowness ($b^{\ast}$), and deformation than the other two samples. The addition of bioactive components did not influence the texture properties in any of the samples. Lipid oxidation (thiobarbituric acid reactive substances [TBARS]) and microorganism count (total plate count [TPC]) were significantly higher in the T1 sample than the two other samples, whereas protein degradation (volatile basic nitrogen [VBN]) was higher in the T2 sample than the other samples. Total amino acid content decreased in the T1 and T3 samples as the storage period increased. Consequently, the T3 sample of Alaska Pollack with breast recovered protein from spent laying hens and cordyceps powder (5 g) + conjugated linoleic acid (CLA) was found to have the necessary functionality to be considered for use in making imitation crab sticks.

• Food Science of Animal Resources
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• v.29 no.6
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• pp.659-667
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• 2009

Casein is considered to be the main source of protein in milk; therefore, many studies have been conducted to identify casein-derived bioactive peptides and their physiological effects. Casein is inactive within the parent protein but can be liberated by various proteases and enzymatic hydrolysis during microbial fermentation and gastrointestinal digestion. Once absorbed, casein exhibits different bioavailabilities in the body. Specifically, casein-derived peptides function as angiotensin converting enzyme (ACE) inhibitor in the cardiovascular system; thus, they are expected to reduce and prevent hypertension. Additionally, casein-derived peptides behave as opioid-like peptides in the nervous system, which impacts relaxation. These peptides are also expected to modulate various aspects of immune functions. Finally, caseinophosphopeptide (CPP) and glycomacropeptide (GMP) may exhibit a number of nutritional effects such as the absorption of calcium, iron or zinc. Many studies have been conducted to evaluate casein-derived peptides due to their multifunctional properties and the results of these studies have contributed to the development of a wide variety of functional dairy products. The purpose of this paper was to review the generation of bioactive peptides, their absorption and metabolism, and their specific bioactive effects.

• Journal of Microbiology and Biotechnology
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• v.19 no.5
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• pp.502-510
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• 2009

Although the Escherichia coli heat-labile enterotoxin B subunit (LTB) has already been expressed in several different systems, including prokaryotic and eukaryotic organisms, studies regarding the synthesis of LTB into oligomeric structures of pentameric size in the budding yeast Saccharomyces cerevisiae have been limited. Therefore, this study used a functional signal peptide of the amylase 1A protein from rice to direct the yeast-expressed LTB towards the endoplasmci reticulum to oligomerize with the expected pentameric size. The expression and assembly of the recombinant LTB were confirmed in both the cell-free extract and culture media of the recombinant strain using a Western blot analysis. The binding of the LTB pentamers to intestinal epithelial cell membrane glycolipid receptors was further verified using a GM1-ganglioside enzyme-linked inmmunosorbent assay (GM1-ELISA). On the basis of the GM1-ELISA results, pentameric LTB proteins comprised approximately 0.5-2.0% of the total soluble proteins, and the maximum quantity of secreted LTB was estimated to be 3 mg/l after a 3-day cultivation period. Consequently, the synthesis of LTB monomers and their assembly into biologically active aligomers in a recombinant S. cerevisiae strain demonstrated the feasibility of using a GRAS microorganism-based adjuvant, as well as the development of carriers against mucosal disease.