In a previous study, we fractionated crude polysaccharide (AME-CP) with macrophage-stimulating activity from a hot-water extract (AME) of Astragalus membranaceus. AME-CP contained glucose (Glc) as a main component sugar, suggesting that it might be rich in starch-like compounds (SLC). To enhance the immunostimulating activity of AME-CP by pruning SLC rarely known to contribute to activity, hydrolysate (AME-SH) was prepared by digesting with starch-related enzymes, including α-amylase and amyloglucosidase. AME-SH was found to contain substances with molecular weights ranging from 3.9 to 84.4 kDa. These substances were primarily composed of galactose, galacturonic acid, Glc, arabinose, rhamnose, and mannose. AME-SH significantly enhanced the production of macrophage-stimulating factors, including nitric oxide (NO), interleukin (IL)-6, and IL-12, in RAW 264.7 cells compared to AME-CP. Treatment of splenocytes isolated from C3H/HeN mice with AME-SH not only promoted IL-6 secretion, but also induced mitogenic activity. In addition, AME-SH promoted the secretion of hematopoietic growth factors including IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in Peyer's patch (PP) cells and stimulated bone marrow cell proliferation through these PP cells. In conclusion, hydrolysate (AME-SH) digested from AME-CP with starch-related enzymes could be used as a potential immunostimulant.
To utilize pepper (Piper nigrum) as an immunostimulatory agent, we isolated macrophage stimulating polysaccharides from pepper and investigated their macrophage activating activities. Hot-water extracts (HW) of black pepper (BP) and white pepper (WP) were prepared, and their macrophage stimulating activities were evaluated using RAW 264.7 cells. BP-HW significantly promoted the secretion of macrophage stimulating factors such as nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin (IL)-6, and IL-12 compared to WP-HW. When BP and WP-HW were fractionated into crude polysaccharides (CP) and low molecules (LM) by ethanol precipitation, BP-CP demonstrated significantly more potent activity. Furthermore, BP-CP not only induced mRNA gene expression of macrophage activation factors, but also promoted nuclear localization of p65 and c-Jun. In addition, component sugar analysis revealed that glucan-type polysaccharides in BP-CP played a crucial role in macrophage activation. Taken together, these findings suggest that black pepper has industrial applicability not only as a spice, but also as an immunostimulatory functional material.
To increase industrial applicability of Astragalus membranaceus (AM) as immunostimulating materials, hot-water extract (AME) was prepared from AM and fermented with Kimchi-lactic acid bacteria (Lactobacillus sakei & Leuconostoc mesenteroides) to prepare fermented AM-postbiotics (FAME). Although FAME prepared from AM-postbiotics did not show a significant enhancement in macrophage stimulating activity compared to non-fermented AME, crude polysaccharide (FAME-CP) fractionated by EtOH precipitation from FAME showed significantly higher macrophage stimulating activity than AME-CP. Compared to AME-CP, FAME-CP showed dramatic changes in component sugar and molecular weight distribution. FAME-CP was a polysaccharide with a major molecular weight distribution of 113.4 kDa containing Man (44.2%), Glc (19.3%), Gal (10.2%), GalA (10.2%), and Ara (7.4%) as sugar components. FAME-CP with enhanced macrophage stimulatory activity not only increased expression levels of mRNA genes encoding macrophage-activated factors (iNOS, TNF-α, MCP-1, IL-6, and COX-2), but also led the nuclear translocation of activated p65 and c-Jun. In conclusion, crude polysaccharide from AM-postbiotics fermented with lactic acid bacteria could increase industrial applicability as a functional material with enhanced immunostimulating activity than AME-CP.
To produce an intestinal immunomodulatory beverage containing Centella asiatica extract (CAE), three types of CAE-added beverage prototypes were prepared, and their immunomodulatory activities and marker compounds were analyzed. As a result of the cytotoxicity assessment, all the beverages did not show significant toxicity compared to the control group. Next, the immunomodulatory activities of the beverage prototype were evaluated using the inflammatory model of IL-1β-induced intestinal epithelial cell line. All the samples significantly reduced the production of IL-6, IL-8, and MCP-1 in a CAE concentration-dependent manner. In addition, CAE-added beverages inhibited NO, IL-6, and IL-12 production in LPS-induced RAW 264.7 cells. When the major triterpenoids, as marker compounds for the production of CAE-added beverages, were analyzed by HPLC-DAD, only asiaticoside was detected beyond the limit of quantification, while madecassoside, madecassic acid, and asiatic acid were not detected. The amounts of asiaticoside in CAE-added beverage prototypes were confirmed in No. 1 (19.39 μg/mL), 2 (19.25 μg/mL), and 3 (19.98 μg/mL). In conclusion, the results of this study suggested that CAE-added beverage prototypes induced immunomodulatory effects in the intestinal inflammatory cell line models and asiaticoside could be used as a marker compound for CAE-added beverage production.
The domestic Pressurized Heavy Water Reactor (PWHR) nuclear power plant, Wolsong Unit 1, was permanently shut down on December 24, 2019. However, research on decommissioning has mainly focused on Pressurized Water Reactors (PWRs), with a notable absence of both domestic and international experience in the decommissioning of PHWRs. If proper business management such as radiation safety and waste is not performed, it can lead to increased business risks and costs in decommissioning. Therefore, the assessment of waste volume and cost, which provide fundamental data for the nuclear decommissioning process, is a crucial technical requirement before initiating the actual decommissioning of Wolsong Unit 1. Decommissioning radiation-contaminated structures and facilities presents significant challenges due to high radiation levels, making it difficult for workers to access these areas. Therefore, technology development should precede decommissioning process assessments and safety evaluations, facilitating the derivation of optimal decommissioning procedures and ensuring worker safety while enhancing the efficiency of decommissioning operations. In this study, we have developed a program to estimate decommissioning waste amounts for PHWRs, building upon prior research on PWR decommissioning projects while accounting for the specific design characteristics of PHWRs. To evaluate the amount of radioactive waste generated during decommissioning, we considered the characteristics of radioactive waste, disposal methods, packaging container specifications, and the criteria for the transfer of radioactive waste to disposal operators. Based on the derived algorithm, we conducted a detailed design and implemented the program. The proposed program is based on 3D modeling of the decommissioning components and the calculation of the Work Difficulty Factor (WDF), which is used to determine the time weighting factors for each task. Program users can select the cutting and packaging conditions for decommissioning components, estimate waste amount based on the chosen decommissioning method, and calculate costs using time weighting factors. It can be applied not only to PHWRs, but also to PWRs and non-nuclear fields, providing a flexible tool for optimizing decommissioning process.
Decommissioning waste is generated with various types and large quantities within a short period. Concrete, a significant building material for nuclear facilities, is one of the largest decommissioning wastes, which is mixed with aggregate, sand, and cement with water by the relevant mixing ratio. Recently, the proposed treatment method for volume reduction of radioactive concrete waste was proven up to scale-up testing using unit equipment, which involved sequentially thermomechanical and chemical treatment. According to studies, the aggregate as non-radioactive material is separated from cement components with contaminated radionuclides as less than clearance criteria, so the volume of radioactive concrete waste is decreased effectively. However, some supplementation points were presented to commercialize the process. Hence, the process requires efficiency as possible to minimize the interface parts, either by integration or rearranging the equipment. In this study, feasibility testing was performed using integrated heating and grinding equipment, to supplement the possible issue of generated powder and dust during the process. Previously, heat treatment and grinding devices were configured separately for pilot-scale testing. But some problems such as leakage and pipe blockage occurred during the transportation of generated fine powder, which caused difficulties in maintaining the equipment. For that reason, we studied to reduce the interface between the equipment by integrating and rearranging the equipment. To evaluate the thermal grinding performance, the fraction of coarse and concrete fines based on 1mm particle size was measured, and the amount of residual cement in each part was analyzed by wet analysis using 4M hydrochloric acid. The result was compared with previous studies and the thermomechanical equipment could be selected to enhance the process. Therefore, it is expected that the equipment for commercialization could be optimized and composed the process compactly by this study.
This study was aimed to isolate bacterial inoculants producing chitinase and evaluate their application effects on corn silage. Four corn silages were collected from four beef cattle farms to serve as the sources of bacterial inoculants. All isolates were tested against Fusarium graminearum head blight fungus MHGNU F132 to confirm their antifungal effects. The enzyme activities (carboxylesterase and chitinase) were also measured to isolate the bacterial inoculant. Based on the activities of anti-head blight fungus, carboxylesterase, and chitinase, L. buchneri L11-1 and L. paracasei L9-3 were subjected to silage production. Corn forage (cv. Gwangpyeongok) was ensiled into a 10 L mini silo (5 kg) in quadruplication for 90 days. A 2 × 2 factorial design consists of F. graminearum contamination at 1.0104 cfu/g (UCT (no contamination) vs. CT (contamination)) and inoculant application at 2.1 × 105 cfu/g (CON (no inoculant) vs. INO (inoculant)) used in this study. After 90 days of ensiling, the contents of CP, NDF, and ADF increased (p<0.05) by F. graminearum contamination, while IVDMD, acetate, and aerobic stability decreased (p<0.05). Meanwhile, aerobic stability decreased (p<0.05) by inoculant application. There were interaction effects (p<0.05) on IVNDFD, NH3-N, LAB, and yeast, which were highest in UCT-INO, UCT-CON, CT-INO, and CT-CON & INO, respectively. In conclusion, this study found that mold contamination could negatively impact silage quality, but isolated inoculants had limited effects on IVNDFD and yeast.
Centella asiatica (C. asiatica) has been widely used in food, cosmetics, and pharmaceutical industry as a functional material. In a previous study, we have investigated not only pharmacological effects such as antioxidative and anti-inflammatory effects, but also analyzed various functional ingredients. In this study, triterpenoids were analyzed using HPLC-DAD to determine marker compounds among functional ingredients. When triterpenoids were analyzed, asiaticoside from C. asiatica was determined as an optimal marker compound. Next, specificity, linearity, limited of detection (LOD), limited of quantification (LOQ), precision, accuracy, and range were evaluated using HPLC-DAD to determine asiaticoside contents in C. asiatica juice and extracts. The specificity was elucidated by chromatogram and retention time using an established analytical method. The coefficient of correlation obtained was 0.9996. LOD was 4.99 μg/mL and LOQ was 15.12 μg/mL. Intra- and inter-day precision of asiaticoside were determined to be 0.48~1.68% and 0.08~1.09%, respectively. Furthermore, the recovery rate of asiaticoside was 98.88% and the analytical range of Field-70E was determined to be 0.625~10 mg/mL. As a results of evaluating ABTS, DPPH, and FRAP antioxidative effect, Field-70E showed potent antioxidant activities. Results of this study could be used as basic data for quality standardization of C. astiatica juice and extracts.
To investigate the anti-inflammatory activity of submerged culture using Cordyceps militaris mycelium, culture-including mycelia was extracted and lyophilized into postbiotics (hot-water extract; CM-HW). HW was fractionated into crude polysaccharide (CM-CP) by ethanol precipitation, and CM-CP was further dialyzed into CM-DCP by dialysis with running water using 12~14 kDa dialysis tube. When the cytotoxicity of subfractions against cells was assessed, no subfraction had a cytotoxic impact that was substantially different from the control groups. In an inflammatory model using LPS-stimulated RAW 264.7 cells, CM-DCP significantly decreased IL-6 and MCP-1 production levels compared to the LPS-control group. CM-DCP also inhibited IL-6 and IL-8 secretion in HaCaT keratinocytes stimulated with TNF-α and IFN-γ. In the meanwhile, the neutral sugar content and mannose ratio of anti-inflammatory CM-DCP were higher than the other fractions, and CM-DCP contained β-1,3/1,6-glucan of 216.1 mg/g. High pressure size exclusion chromatography revealed that CM-DCP contained molecules with a molecular weight range of 5.6 to 144.0 kDa. In conclusion, postbiotics of C. militaris mycelium significantly promoted anti-inflammatory activity, suggesting that neutral polysaccharides including Glc and Man contribute to the anti-inflammation in RAW 264.7 or HaCaT cells.
To enhance the bioavailability and bioactivities of mixed herbal medicines (RW), they were fermented with lactic-acid bacteria isolated from kimchi into postbiotics (FRW). Then, from the results of the 16s rRNA sequencing analysis, lactic acid bacteria isolated from kimchi were identified to be of two species, namely Lactobacillus sakei and Leuconostoc mesenteroides. The FRW prepared from the RW were extracted using hot water (HW) and 70% EtOH (EtOH) for comparison of their macrophage-stimulating activities. Based on a comparison of the activities of the FRW extracts, nitric oxide (NO) production of HW was significantly higher than that in EtOH. An analysis of the chemical properties of the extracts showed that HW had higher contents of neutral sugar and uronic acid than EtOH as well as contained a large amount of glucose. In addition, crude polysaccharide (CP) was prepared to enhance the macrophage-stimulating activity. The FRW-CP not only secreted immunostimulatory mediators but also increased the expression of immunostimulatory genes (iNOS, TNF-α, MCP-1, and IL-6). The fractionated FRW-CP contained about 90% neutral sugars, and these sugars were mainly composed of glucose, galacturonic acid, and arabinose. Thus, FRW prepared by fermentation of RW with kimchi lactic acid bacteria were found to be immunostimulatory modulators.
With the aging of nuclear power plants (NPPs) in 37 countries around the world, 207 out of 437 NPPs have been permanently shutdown as of August 2022 according to the IAEA. In Korea, the decommissioning of NPPs is emerging as a challenge due to the permanent shutdown of Kori Unit 1 and Wolsong Unit 1. However, there are no cases of decommissioning activities for Heavy Water Reactor (HWR) such as Wolsong Unit 1 although most of the decommissioning technologies for Light Water Reactor (LWR) such as Kori Unit 1 have been developed and there are cases of overseas decommissioning activities. This study shows the development of a decommissioning waste amount/cost/process linkage program for decommissioning Pressurized Heavy Water Reactor (PHWR), i.e. CANDU NPPs. The proposed program is an integrated management program that can derive optimal processes from an economic and safety perspective when decommissioning PHWR based on 3D modeling of the structures and digital mock-up system that links the characteristic data of PHWR, equipment and construction methods. This program can be used to simulate the nuclear decommissioning activities in a virtual space in three dimensions, and to evaluate the decommissioning operation characteristics, waste amount, cost, and exposure dose to worker. In order to verify the results, our methods for calculating optimal decommissioning quantity, which are closely related to radiological impact on workers and cost reduction during decommissioning, were compared with the methods of the foreign specialized institution (NAGRA). The optimal decommissioning quantity can be calculated by classifying the radioactivity level through MCNP modeling of waste, investigating domestic disposal containers, and selecting cutting sizes, so that costs can be reduced according to the final disposal waste reduction. As the target waste to be decommissioning for comparative study with NAGRA, the calandria in PHWR was modeled using MCNP. For packaging waste container, NAGRA selected three (P2A, P3, MOSAIK), and we selected two (P2A, P3) and compared them. It is intended to develop an integrated management program to derive the optimal process for decommissioning PHWR by linking the optimal decommissioning quantity calculation methodology with the detailed studies on exposure dose to worker, decommissioning order, difficulty of work, and cost evaluation. As a result, it is considered that it can be used not only for PHWR but also for other types of NPPs decommissioning in the future to derive optimal results such as worker safety and cost reduction.
Maca roots (Lepidium meyenii) are an important medicinal herb that have long been used by the Andes-indigenous peoples and South Americans. In Korea, recently, it has attracted attention as a health food material because of nutritional values and physiological activities. The purpose of this study was to investigate the industrial applicability of maca (red and golden varieties; R&G) as immunostimulating materials. In the macrophage stimulating assay using RAW 264.7 cells at 125~500 μg/mL of non-cytotoxicity doses, G-HW showed the most potent production of TNF-α, IL-6 and nitric oxide compared to red maca or the other extracts. The general component analysis results showed that all extracts comprised more than 90% neutral sugars with small amounts of uronic acid and protein. Meanwhile, component sugar analysis showed the difference in the content of uronic acids of cold- and hot-water extract. Additionally, the further fractionation of G-HW into crude polysaccharide (G-CP) greatly enhanced the macrophage stimulating activity, and G-CP contained macromolecules over 144 kDa, comprised mainly of glucose and galacturonic acid (51.0 and 34.9%). In conclusion, crude polysaccharide from maca showed industrial applicability as immunostimulating material, and especially golden maca showed higher macrophage stimulating activity than red maca.
Compound K (20-O-β-(D-glucopyranosyl)-20(S)-protopanaxadiol)는 진세노사이드의 활 성성분이다. Compound K는 경구 투여 후 Rb1, Rb2 및 Rc가 사람의 장내 미생물의 β-glucosidase에 의 해 생물전환 과정을 거쳐 생성된다고 알려져 있다. 본 연구는 생물전환된 인삼농축액에서 얻은 compound K를 이용해 항염증 및 독성을 조사하였다. 세포독성평가 결과, compound K는 0.001∼1 μg/mL의 농도 범위에서 유의적인 세포독성은 나타나지 않았으며, LPS로 염증이 유발된 RAW 264.7 세포에서 TNF-α, MCP-1, IL-6 및 NO의 생성을 억제하는 것으로 확인되었다. 동일 농도범위에서 TNF-α 및 IFN-γ로 염증이 유발된 HaCaT 세포는 compound K의 처리로 인해 IL-8의 생성을 감소시키는 것으로 나타났지만, IL-6의 경우 일부 농도에서 생성을 감소시켰으나, 통계적인 유의성은 나타나지 않았다. Brine shrimp를 이 용한 치사율 검정법에서 compound K의 LC50는 0.37mg/mL로 다소의 독성을 함유하고 있는 것으로 나타 났으나 compound K가 35% 고함유된 생물전환물은 LC50가 0.87mg/mL로 나타나 상대적으로 낮은 독성 을 보였다. 따라서 이 생성물은 향후 여드름 완화용 화장품 개발에 사용할 수 있는 매우 우수한 기능성 소 재가 될 수 있을 것으로 기대된다.
To investigate the industrial availability of liquid fermentation (PL-ferment) by Phellinus linteus mycelium as a postbiotics for the inhibition of inflammation, PL-ferment was fractionated into culture supernatant (CS), hot-water extract (HW) from PL-ferment, EtOH-precipitate (CP) fractionated from HW, and the dialysate (DCP) of CP. Compared to the other fractions, DCP which is expected to contain exopolysaccharide (EPS) as the major component, significantly decreased the production of NO, IL-6, and MCP-1 in LPS-induced RAW 264.7 cells, and IL-6 and IL-8 in TNF-α and IFN-γ-induced HaCaT cells. The general component analysis results showed that no significant difference in components was observed between the fractions, whereas sugar composition analysis revealed that DCP had decreased glucose and increased mannose contents compared to the other fractions. This suggests that mannose played an important role in the anti-inflammatory activity of the active fraction, DCP. Molecular weight distribution analysis revealed that DCP was mainly composed of low-molecular-weight material-removed high-molecular-weight polysaccharides of 18–638 kDa, suggesting that EPS originated from P. linteus EPS. In conclusion, our results suggest that the DCP of P. linteus mycelium fermentation using the anti-inflammatory activity could be used industrially as postbiotic material.
To utilize Malus pruniforia Borkh. as a functional material, cold-water (CW), hot-water (HW), and 70% ethanol (EtOH) extracts were prepared, and their antioxidant and anti-inflammatory activities were compared. The antioxidant activity of the HW extract evaluated by ABTS and DPPH radical scavenging and FRAP activity was significantly effective. The total polyphenol content of the HW extract was also higher by 15.5±0.7 mg GAE/g extract compared to other extracts. The EtOH extract showed significantly decreased TNF-α (39.8%), IL-6 (65.5%), and NO (34.9%) levels in RAW 264.7 cells compared to the LPS-induced control group. The levels of IL-6 (21.1%) and IL-8 (19.3%) were significantly decreased by treatment of EtOH extract in HaCaT keratinocytes induced with TNF-α and IFN-γ. The UHPLC-MS results indicated that the EtOH extract might have chlorogenic acid and phlorizin as the major compounds. This was validated using HPLC-DAD, which showed that the EtOH extract had higher levels of chlorogenic acid and phlorizin (1,185±58 and 470±10 μg/g extract, respectively). In conclusion, the present study suggested that the anti-inflammatory activity of the EtOH extract was more effective than the CW and HW extracts, and chlorogenic acid and phlorizin could be used as indicator compounds and functional substances.
Many transcription factors are involved in directing the growth of porcine oocytes. The localization and expression level of a given transcription factor often differ at each stage of early embryonic growth, which spans from fertilization to the formation of the blastocyst. A hallmark of the blastocyst stage is the separation of the endodermal and mesodermal ectoderm. The embryo's medium and its effects are known to be crucial during early development compared to the other developmental stages, and thus require a lot of caution. Therefore, in many experiments, early development is divided into the quality of oocyte and cumulus cells and used in experiments. We thought that we were also heavily influenced by genetic reasons. Here, we examined the expression patterns of five key transcription factors (CDX2, OCT4, SOX2, NANOG, and E-CADHERIN) during porcine oocyte development whose expression patterns are controversial in the pig to the literature. Antibodies against these transcription factors were used to determine the expression and localization of them during the early development of pig embryos. These results indicate that the expressions of key transcription factors are generally similar in mouse and pig early developing embryos, but NANOG and SOX2 expression appears to show species-specific differences between pig and mouse developing embryos. This work helps us better understand how the expression patterns of transcription factors translate into developmental effects and processes, and how the expression and localization of different transcription factors can crucially impact oocyte growth and downstream developmental processes.
Pigs are considered an ideal source of human disease model due to their physiological similarities to humans. However, the low efficiency of in vitro embryo production (IVP) is still a major barrier in the production of pig offspring with gene manipulation. Despite ongoing advances in the associated technologies, the developmental capacity of IVP pig embryos is still lower than that of their in vivo counterparts, as well as IVP embryos of other species (e.g., cattle and mice). The efficiency of IVP can be influenced by many factors that affect various critical steps in the process. The previous relevant reviews have focused on the in vitro maturation system, in vitro culture conditions, in vitro fertilization medium, issues with polyspermy, the utilized technologies, etc. In this review, we concentrate on factors that have not been fully detailed in prior reviews, such as the oocyte morphology, oocyte recovery methods, denuding procedures, first polar body morphology and embryo quality.