• Journal of Plant Biotechnology
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• v.48 no.3
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• pp.156-164
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• 2021

Spirodela polyrhiza, from the Lemnaceae family, are small aquatic plants that offer an alternative plant-based system for the expression of recombinant proteins. However, no turion transformation protocol has been established in this species. In this study, we exploited a pB7YWG2 vector harboring the eYFP gene that encodes enhanced yellow fluorescent protein (eYFP), which has been extensively used as a reporter and marker to visualize recombinant protein localization in plants. We adopted Agrobacterium tumefaciens-mediated turion transformation via vacuum infiltration to deliver the eYFP gene to turions, special vegetative forms produced by duckweeds to endure harsh conditions. Transgenic turions regenerated several duckweed fronds that exhibited yellow fluorescent emissions under a fluorescence microscope. Western blotting verified the expression of the eYFP protein. To the best of our knowledge, this is the first report of an efficient protocol for generating transgenic S. polyrhiza expressing eYFP via Agrobacterium tumefaciens-mediated turion transformation. The ability of turions to withstand harsh conditions increases the portability and versatility of transgenic duckweeds, favoring their use in the further development of therapeutic compounds in plants.

• Journal of Dairy Science and Biotechnology
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• v.41 no.1
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• pp.34-43
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• 2023

Hydrolysis of whey-derived proteins using lactic acid bacteria (LAB) utilizes the mass culture method and fermentation of LAB to produce effective bioactive peptides. Whey protein has the biological potential of its precursors, but the active fragments may not be released depending on the hydrolysis method. As an alternative to these problems, the nutritional and bioactive functionality of the hydrolysis method have been reported to be improved using LAB for whey protein. Peptide fractions were obtained using a sample fast protein liquid chromatography device. Antioxidant activity was verified for each of the five fractions obtained. In vitro cell experiments showed no cytotoxicity and inhibited nitric oxide production. Cytokine (IL [interleukin]-1α, IL-6, tumor necrosis factor-α) production was significantly lower than that of lipopolysaccharides (+). As a result of checking the amino acid content ratio of the fractions selected through the AccQ-Tag system, 17 types of amino acids were identified, and the content of isoleucine, an essential amino acid, was the highest. These properties show their applicability for the production of functional products utilizing dietary supplements and milk. It can be presented as an efficient method in terms of product functionality in the production of uniform-quality whey-derived peptides.

• Journal of Periodontal and Implant Science
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• v.45 no.2
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• pp.69-75
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• 2015

Purpose: Salivary fluid formation is primarily driven by Ca2+-activated, apical efflux of chloride into the lumen of the salivary acinus. The anoctamin1 protein is an anion channel with properties resembling the endogenous calcium-activated chloride channels. In order to better understand the role of anoctamin proteins in salivary exocrine secretion, the expression of the ten members of the anoctamin gene family in the mouse submandibular gland was studied. Methods: Total RNA extracted from mouse submandibular salivary glands was reverse transcribed using primer pairs to amplify the full-length coding regions of each anoctamin gene and was subcloned into plasmid vectors for DNA sequencing. Alternative splice variants were also screened by polymerase chain reaction using primer pairs that amplified six overlapping regions of the complementary DNA of each anoctamin gene, spanning multiple exons. Results: Multiple anoctamin transcripts were found in the mouse submandibular salivary gland, including full-length transcripts of anoctamin1, anoctamin3, anoctamin4, anoctamin5, anoctamin6, anoctamin9, and anoctamin10. Exon-skipping splicing in the N-terminal exons of the anoctamins1, anoctamin5, and anoctamin6 genes resulted in multiple alternative splice variants. No expression of anoctamin2, anoctamin7, or anoctamin8 was found. Conclusions: The predominant anoctamin transcript expressed in the mouse submandibular gland is anoctamin1ac. The chloride channel protein produced by anoctamin1ac is likely responsible for the $Ca^{2+}$-activated chloride efflux, which is the rate-limiting step in salivary exocrine secretion.

• Journal of Crop Science and Biotechnology
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• v.11 no.2
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• pp.127-134
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• 2008

Tobamoviruses are the major viral pathogens of tomato and bell pepper. The preliminary results showed that Acibenzolar-Smethyl(ASM; S-methylbenzo(1,2,3) thiadiazole-7-carbothiate) pre-treatment to tomato and tobacco plants reduces the concentration of Tomato mosaic tobamovirus(ToMV) and Tobacco mosaic tobamovirus(TMV) in tomato and bell pepper seedlings, respectively. Pre-treatment of the indicator plant(Nicotiana glutinosa) with the ASM followed by challenge inoculation with tobamoviruses produced a reduced number and size of local lesions(67 and 79% protection over control to TMV and ToMV inoculation, respectively). In order to understand the mechanism of resistance the gene expression profiles of antiviral genes was examined. RT-PCR products showed higher expression of two viral resistance genes viz., alternative oxidase(AOX) and RNA dependent RNA polymerase(RdRp) in the upper leaves of the ASM-treated tomato plants challenge inoculation with ToMV. Further, the viral concentration was also quantified in the upper leaves by reverse transcription PCR using specific primer for movement protein of ToMV, as well as ELISA by using antisera against tobamoviruses. The results provided additional evidence that ASM pre-treatment reduced the viral movement to upper leaves. The results suggest that expressions of viral resistance genes in the host are the key component in the resistance against ToMV in the inducer-treated tomato plants.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.571-577
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• 1990

Alkali extract obtained from mycelia of Lentinus edodes IY105 was shown to have potent anticomplementary activity and alternative complementary activity in vitro. It was also shown to activate reticuloendothelial system of ICR mice in the carbon clearance. Amount of carbohydrates and protein of the extract were 17% and 42% respectively. It was found that carbohydrates were consisted of 5 monosaccharides and protein was consisted of 16 amino acids. Antitumor activity was observed in the alkali extract treated group of sarcoma 180 bearing ICR mice.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.1
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• pp.89-93
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• 2002

A total of six crossbred barrows ($Duroc{\times}Landrace{\times}Large$ White, $44.17{\pm}1.94kg$ BW) were housed conducted to evaluate apparent fecal digestibilities of Brown Rice (BR) as an alternative energy source in growing pigs. Pigs were housed individually on metabolism crate on the basis of body weight. Four treatments contained: 1) 100% of corn-soybean meal (C100; Control diet), 2) 75% of corn-soybean meal diet plus 25% of corn meal (C25), 3) 100% of brown rice-soybean meal diet (BR100), 4) 75% of brown rice-soybean meal diet plus 25% of brown rice meal (BR25). Brown rice has an excellent gross energy and crude protein composition compared to corn. The BR used had 3,801 kcal of gross energy/kg, 8.0% crude protein, 2.6% of ether extract, 0.035% calcium and 0.35% total phosphorus. The best digestibilities of energy (87.75%), DM (81.71%) and CP (78.57%) were observed in BR 100 group and the worst were found in Corn 25 group. The nutrient digestibility was not significantly different in most nutrients. Through this experiment, BR appeared a good alternative energy source that can replace corn yellow to 100% in growing pigs. Therefore, the price relationship between corn and BR may provide an excellent opportunity for pork producers to use BR in order to reduce feed costs provided that diet has been balanced for digestible amino acids.

• Animal Bioscience
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• v.36 no.7
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• pp.1091-1100
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• 2023

Objective: The present study is to examine the effect of high inclusion of co-products in pig diets (referred to as an alternative diet) during the finishing stage on pig growth performance, meat quality and boar taint compounds. Methods: Growing pigs were fed an alternative diet made with distillers dried grains with solubles (25%), canola meal (20%), and wheat middling (15%) or a control diet based on barley and soybean meal to investigate the impact of co-products on pig performance and meat quality. Sixteen female and sixteen entire male Duroc×(Large White×Landrace) pigs (22.6±2.07 kg, body weight±standard error) were equally allocated to the diets. Results: Pigs fed the alternative diet had a lower feed intake; however, growth rate and feed conversion efficiency were unaffected by diet. A diet by sex interaction was found for gain:feed whereby males fed the alternative diet had the best feed conversion (p<0.01). Pork from pigs fed the alternative diet had lower a^* and Chroma and protein % (p<0.05), while other meat quality characteristics were unaffected. The alternative diet reduced backfat skatole levels (p<0.001). Conclusion: A diet containing high inclusion levels of co-products can be fed to pigs during the finishing stage without detrimental effects on pig performance or meat quality and with the potential to enhance pork flavour. This finding suggests a solution to increase the sustainable development of pig production.

• Applied Microscopy
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• v.31 no.2
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• pp.199-206
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• 2001

In 1980s, the fragmentation or subdivision of protein deposits at the periphery of protein storage vacuole was suggested as the only route of PB development in pea cotyledon cells. Since then, other independant processes such as terminal dilation , transformation and de novo development have been discussed as alternative routes for PB development, and today, these multiple mechanisms of PB development are accepted as a result of active investigations. For analysis of the protein accumulations in the ER cisternae during seed development, immunocytochemical gold labellings were applyed on the single cells separated by enzymatic digestion from cotyledon tissue. Anti-legumin labellings at the early stage, and anti-vicilin labellings at the intermediate stage were observed on the protein-filled ER. The $\alpha-Tip$, which is the ER retention protein, was labelled somewhat at late stage, and PPase, a sort of tonoplast membrane protein, was labelled at early stage.

• Kidney Research and Clinical Practice
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• v.37 no.4
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• pp.384-392
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• 2018

Background: A very low protein diet (VLPD) with ketoacid analogs of essential amino acids (KA/EAA) administration can remarkably influence protein synthesis and metabolic disturbances of patients with advanced chronic kidney disease (CKD), and may also slow the decline in renal function. Methods: A retrospective cohort study was carried out to monitor renal progression and metabolic and nutritional status among 140 patients with CKD stage III or IV. One group (n = 70) was on a low protein diet (LPD) with 0.6 g of protein intake, and another group (n = 70) was on a VLPD with 0.3 g of protein and KA/EAA supplementation of 100 mg/kg/day for 12 months. Results: At 12-month follow-up, estimated glomerular filtration rate (GFR) significantly decreased from $41.6{\pm}10.2$ to $36.4{\pm}8.8mL/min/1.73m^2$ (P < 0.001) and urine protein increased from $0.6{\pm}0.5$ to $0.9{\pm}1.1g/day$ (P = 0.017) in the LPD group, but no significant changes in estimated GFR and urine protein were found in the VLPD plus KA/EAA group. A significant mean difference in rate of change in estimated GFR ($-5.2{\pm}3.6mL/min/1.73m^2$ per year; P < 0.001) was observed between the two groups. After Cox regression analysis, treatment with VLPD plus KA/EAA significantly protected against the incidence of declining GFR > 10% annually (adjusted hazard ratio, 0.42; 95% confidence interval, 0.23-0.79; P = 0.006) and significant correlations were found between using VLPD plus KA/EEA and increased GFR. Conclusion: VLPD supplementation with KA/EAA is associated with delayed renal progression while preserving the nutritional status in the patients with CKD. Co-administration of VLPD and KA/EAA may prove an effective alternative to conservative management of CKD.

• Development and Reproduction
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• v.17 no.4
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• pp.435-440
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• 2013

Previous studies, including our own, have demonstrated that the intratesticular injection of hypertonic saline (20%) decreased serum testosterone level which was similar to the surgical castration in the rat, showing the state of chemical castration. In the present study, we further verify the efficacy of this less invasive method as an alternative of surgical orchidectomy in the andrological field. Sterilized 20% saline was directly injected into the adult male rats (750 ${\mu}l$ per testis). The tested rats were divided into 3 groups including intact group (intact), orchidectomy group (ORX) and saline injection group (SAL) after bilateral orchidectomy was performed at the same day of injection. All rats were sacrificed at 4 weeks after injection. The reproductive organs (testes, epididymis, seminal vesicles and prostates) were collected and used for DNA and protein pattern analyses. Also, patho-histological studies on the testes were performed. In contrast to the intact group, similar DNA damages of testis and seminal vesicle were appeared in ORX group and SAL group. The DNA degradations seemed to be the results of necrosis rather than apoptosis. In the protein pattern analysis, all the testing tissues exerted similar patterns in the ORX group and the SAL group compared to the those of intact group. Patho-histological studies revealed that severe degenerative changes in testicular seminiferous tubules and massive infiltration of immune cells in SAL group. The present study confirmed that direct injection of hypertonic saline into the testis caused the equivalent biochemical changes in the accessory sex organs as shown in the orchidectomized animals. These results suggest that hypertonic saline injection model could be a useful castration model which can substitute for surgical castration when its safety is secured through further study in the future.