• Journal of Microbiology and Biotechnology
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• v.22 no.11
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• pp.1575-1579
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• 2012

Paenibacillus polymyxa is known to be a plant-growth-promoting rhizobacterium. The present study describes a quantitative polymerase chain reaction (qPCR) assay for the specific detection and quantitation of P. polymyxa using a primer pair based on the sequence of a membrane-fusion protein for the amplification of a 268 bp DNA fragment. This study reports that the qPCR-based method is applicable for the rapid and sensitive detection of P. polymyxa and can be used as an alternative method for agricultural soil monitoring.

• Journal of Chest Surgery
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• v.21 no.4
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• pp.613-618
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• 1988

The exposure of blood to foreign materials can cause the denaturation of plasma protein components such as immunoglobulins and complements. And those phenomena increase the morbidity and mortality after intracardiac operations through the cardiopulmonary bypass. From April, 1987 to September, 1987, we had observed the serial changes of plasma total protein IgG, IgA, IgM, complements[C3, C4] in bubble oxygenator group[n=5] and membrane oxygenator group[n=5]. Statistically significant difference between two groups were present in total protein and C3. We conclude that using membrane oxygenator in long extracorporeal circulation can reduce the activation of alternative pathway of complement system, and which can reduce post-perfusion complications of the lung though we can`t prove it in mass populations.

• Biomolecules & Therapeutics
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• v.20 no.3
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• pp.340-345
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• 2012

Sarsasapogenin (SAR) is a steroidal sapogenin that is used as starting material for the industrial synthesis of steroids. It has various pharmacological benefits, such as antitumor and antidepressant activities. Since its effect on melanin biosynthesis has not been reported, we used murine melanocyte melan-a cells to investigate whether SAR influences melanogenesis. In this study, SAR significantly increased the melanin content of the melan-a cells from 1 to 10 ${\mu}M$. Based on an enzymatic activity assay using melan-a cell lysate, SAR had no effect on tyrosinase and DOPAchrome tautomerase activities. It also did not affect the protein expression of tyrosinase-related protein 1 and DOPAchrome tautomerase. However, protein levels of tyrosinase and microphthalmia-associated transcription factor were strongly stimulated by treatment with SAR. Therefore, our reports suggest that SAR treatment may induce melanogenesis through the stimulation of tyrosinase and microphthalmia-associated transcription factor expression in melan-a cells.

• Environmental Engineering Research
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• v.20 no.2
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• pp.163-169
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• 2015

In this study, we attempted to solubilize protein in slaughter blood (SB) using ultrasonic technology. The application of ultrasonic technology can make enzymatic degradation of SB more effective, which has no comparable alternative for treatment. The SB was homogenized by grinding it for 10 minutes at 10,000 rpm as a pretreatment for preventing its clotting, and then ultrasonic treatment was attempted to solubilize protein in SB. To maximize the efficiency of ultrasonic treatment for SB, the optimum condition of ultrasonic frequency (UF) was determined to be 20 kHz. To optimize the operation conditions of ultrasonification with 20 kHz of frequency, we used response surface methodology (RSM) based on ultrasonic density (UD) and ultrasonification time (UT). The solubilization rate (SR) of protein (%) was calculated to be $101.304-19.4205X_1+0.0398X_2+7.9411X_1{^2}+0.0001X_2{^2}+0.0455X_1X_2$. From the results of the RSM study, the optimum conditions of UD and UT were determined at 0.5 W/mL and 22 minutes, respectively, and SB treated under these conditions was estimated to have a 95% SR. Also, experimentally, a 95.53% SR was observed under same conditions, accurately reflecting the theoretical prediction of 95%.

• International Journal of Industrial Entomology and Biomaterials
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• v.32 no.2
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• pp.49-53
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• 2016

Bone morphogenetic proteins (BMPs) are essential growth factors for bone formation, skeletal development and bone regeneration. The BMP-2/7 heterodimer is known to have remarkable effects on osteogenic induction that are even stronger than the BMP-2 or BMP-7 homodimers. We designed a recombinant human BMP-2/7 (rhBMP-2/7) heterodimer protein with four glycine residues between BMP-2 and BMP-7 protein to facilitate free bond rotation of domains. The Baculovirus Expression Vector System (BEVS) is routinely used to produce recombinant proteins in the milligram scale. In this study, the BEVS was used to express the rhBMP-2/7 protein whrer the recombinant baculovirus was recovered in the host Sf9 cells. To confirm the biological activity of rhBMP-2/7 protein secreted from the BEVS as an osteogenic differentiation and induction factor, we measured the BMP-induced ALP activity. rhBMP-2/7 could be used as an alternative to BMPs to overcome limitations like short half-life and requirement for high concentrations. Furthermore, rhBMP-2/7 may be an efficient tool for various application studies such as bone regeneration and skeletal development.

• 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.