Chicken embryonic stem (ES) cells have great potential and provide a powerful tool to investigate embryonic development and to manipulate genetic modification in a genome. However, very limited studies are available on the functional characterization and robust expansion of chicken ES cells compared to other species. Here, we have developed a method to generate chicken embryonic stem cell-like cells under pluripotent culture conditions. The chicken embryonic stem cell-like cells were cultivated long-term over several passages of culture without loss of pluripotency in vitro and had the specific expression of key stem cell markers. Furthermore, they showed severe changes in morphology and a significant reduction in pluripotent genes after siRNA-mediated NANOG knockdown. Collectively, these results demonstrate the efficient generation of chicken embryonic stem cell-like cells from EGK stage X blastoderm-derived singularized cells and will facilitate their potential use for various purposes, such as biobanking genetic materials and understanding stemness in the fields of animal biotechnology.

세리신은 누에고치에서 추출한 단백질로 많은 건강상의 이점을 가지고 있다. 본 연구는 화장품 소재로서 누에고치에서 유래된 세리신 표준품의 항주름 활성 및 항염증 활성을 평가하기 위해 수행되었다. 세리신의 항산화 효과는 DPPH 및 ABTS 측정법에 의해 측정되었다. 또한 대식세포인 Raw 264.7 cell에서 의 세포 생존율을 확인하였으며, lipoplyscaccharide를 이용하여 유도된 염증반응을 이용하여 세리신의 항 염증 효과를 조사하였다. 그 결과 세리신은 DPPH, ABTS에서 항산화 활성을 보였으며 세포 독성을 가지지 않은 1,000 μg/mL의 농도에서 NO를 억제하였다. 종합하여 세리신은 항산화 활성을 가지며 항노화 및 항 염증 화장품의 우수한 소재가 될 수 있음을 보여준다.

The present study was designed to investigate the antiproliferative activity and molecular mechanisms of Bibimbap in HT-29 human colorectal adenocarcinoma cells. Bibimbap extract inhibited the proliferation of HT-29 cells by 50% at a concentration of 10.1±0.17 mg/mL for 48 h. The population of live cells decreased slightly, and the morphology changed with a reduction in cell volume (pyknosis) with Bibimbap. Treatment with 5 mg/mL of Bibimbap resulted in slight cell shrinkage. Furthermore, as the Bibimbap dose increased to 10 mg/mL, these characteristics were more evident, and HT-29 cells exhibited partial detachment by staining with the DNA-binding dye Hoechst 33342. Flow cytometric analysis by Annexin V and PI double staining showed that Bibimbap increased the levels of apoptosis. Analysis of the mechanism of these events showed that Bibimbap-treated cells exhibited a mitochondria-dependent apoptotic pathway through the modulation of caspase-3, caspase-8, caspase-9, and poly-ADP ribose polymerase, as well as Bax and Bcl-2 expression in dose- and time-dependent manners. Consequently, Bibimbap exerts a significant antiproliferative effect on HT-29 human colorectal adenocarcinoma cells.

Bis(2-ethylhexyl) phthalate (DEHP) is one of the plasticizers used in the polyvinyl chloride (PVC) industry. It is known to be easily released into the environment. In this study, we investigated effects of DEHP on growth, metabolic pathway, and virulence gene expression in soil-borne bacterial plant pathogen, Pectobacterium carotovorum SCC1 using in vitro assays. As a result, DEHP at 20 μg mL-1 did not affect the growth, cell membrane permeability, or ATPase activity of P. carotovorum SCC1. However, it decreased succinyl-CoA synthase (SCS) activity in the tricarboxylic acid (TCA) cycle. Relative expression levels of virulence genes encoding pectate lyase and pectin were differentially influenced by DEHP treatment. These results suggest that biological characteristics of P. carotovorum might be influenced by DEHP in soil.

In 1951, Colin Russell Austin and Min Chueh Chang identified “capacitation”, a special process involving ejaculated spermatozoa in the female reproductive tract. Capacitation is a phenomenon that occurs in vivo , but almost all knowledge of capacitation has been obtained from in vitro studies. Therefore, numerous trials have been performed to establish in vitro capacitation methods for various studies on reproduction. Although a series of studies have been conducted to develop an optimal protocol for inducing capacitation, most have focused on identifying the appropriate chemical compounds to induce the capacitation of boar spermatozoa in vitro. Therefore, the purpose of this study was to identify the optimal incubation time for inducing capacitation in vitro. Duroc semen was incubated for various periods (60, 90, and 120 min) to induce capacitation. Sperm function (sperm motility, motion kinematic parameters, and capacitation status) was evaluated. The results showed that total sperm motility, rapid sperm motility, progressive sperm motility, curvilinear velocity, and average path velocity significantly decreased in a time-dependent manner. However, the capacitation status did not show any significant changes. Taken together, these results indicate that an incubation time of more than 60 min suppresses sperm motility and motion kinematic parameters. Therefore, we suggest that 60 min may be the best incubation time to induce capacitation without negative effects on sperm motility and motion kinematics in boar spermatozoa in vitro.

This study evaluated the effect of lactic acid bacteria (LAB, a mixture of Enterococcus faecium and Lactobacillus plantarum) supplementation, the storage temperature, and storage period on the fermentation characteristics and in vitro ruminal digestibility of a total mixed ration (TMR). The TMR was prepared into two groups, namely, CON (control TMR without the LAB) and ML (supplementing a mixture of E. faecium and L. plantarum in the ratio of 1% and 2% (v/w), respectively). Both groups were divided and stored at 4°C or 25°C for 3, 7, and 14 d fermentation periods. Supplementing LAB to the TMR did not affect the chemical composition of TMR except for the lactate and acetate concentration. Storage temperatures affected (p<0.05) the chemical composition of the TMR, including pH, lactate, and acetate contents. The chemical composition of TMR was also affected (p<0.05) by the storage period. During in vitro rumen fermentation study, the ML treatment showed lower (p<0.05) dry matter digestibility at 24 h incubation with a higher pH compared to the CON. There was no difference in the in vitro dry matter digestibility (IVDMD) of TMR between the CON and ML treatment however, at 24 h, ML treatment showed lower (p<0.05) IVDMD with a higher pH compared to the CON. The effects of storage temperature and period on IVDMD were not apparent at 24 h incubation. In an in vivo study using Holstein steers, supplementing LAB to the basal TMR for 60 d did not differ in the final body weight and average daily gain. Likewise, the fecal microbiota did not differ between CON and ML. However, the TMR used for the present study did include a commercial yeast in CON, whereas ML did not; therefore, results were, to some extent, compromised in examining the effect of LAB. In conclusion, storage temperature and period significantly affected the TMR quality, increasing acetate and lactate concentration. However, the actual effects of LAB supplementation were equivocal.

Because Cnidium officinale is susceptible to viral infection, this study investigated the proportion of virus elimination and plant growth after treatments with thermotherapy and chemotherapy. In the thermotherapy group, Apple stem grooving virus (ASGV) and Cnidium vein yellowing virus–2 (CnVYV-2) were significantly reduced in 33℃ treatment. However, Cnidium vein yellowing virus–1 (CnVYV-1) was not affected by any of the four temperature treatments, and Cucumber mosaic virus (CMV) did not show the tendency to eliminate viruses according to temperature treatment. In the investigation of in vitro growth characteristics, plants were not significantly affected by temperature treatments. In the chemotherapy group, 40 mg･L-1 ribavirin treatment was the most effective in eliminating viruses. Different ribavirin concentrations (0 to 40 mg･L-1) did not significantly influence the in vitro plant survival rate. ASGV was completely eliminated with combined thermotherapy and chemotherapy (T29R20 and T29R40). Although the combined thermotherapy and chemotherapy treatment did not affect in vitro plant growth, it significantly influenced root development. The results indicate that thermotherapy, chemotherapy, and the combined treatment effectively eliminated these four viruses (ASGV, CnVYV-1, CnVYV-2, and CMV) without producing any major problems in C. officinale.

In vitro digestibility and protein digestibility corrected amino acid scores (PDCAAS) were investigated to verify the availability of protein in various Rhizopus oligosporus fermented products of domestic soybean (Glycine max L.) cultivars. Danbaegkong (DBK), Daepung (DP), Daewonkong (DWK), Saedanbaek (SDB), Seonyu (SY), and Cheongja4ho (CJ4) were used as raw samples, which were fermented using commercially available Rhizopus oligosporus for 48 h. All cultivars showed increased crude protein content after fermentation. The crude protein content of DBK and SDB was significantly higher than that of the other samples (55.12% in DBK and 54.22% in SDB) (p<0.001). CJ4 had the highest alanine content of 28.88 mg/g (p<0.001), and no significant difference in cysteine content was detected among the cultivars. In most of the fermented samples, the in vitro digestibility was 0.9 or higher, indicating high protein in the fermented samples. However, it is considered that restrictions on digestion are low. In DWK, the amino acid content and PDCAAS, which together indicate protein quality, were 0.917 and 0.855, respectively, confirming that it was the best cultivar to provide the raw material for fermentation. In conclusion Rhizopus oligosporus fermented soybean products can be considered a prospective source of protein with high utility value.

In this study, probiotic properties such as acid resistance, bile tolerance, intestinal adhesion and fat accumulation inhibition of lactic acid bacteria isolated from kimchi were investigated in order to develop probiotics effective anti-adipogenic effects. Among the 120 types of lactic acid bacteria isolated from kimchi, 42 types of lactic acid bacteria being to acid resistance, 35 types of lactic acid bacteria being to bile resistance, and 24 types of lactic acid bacteria being to both acid resistance and bile resistance were selected. These 24 types were tested for the intestinal adhesion ability which is an essential factor of probiotics’ function and L. rhamnosus GG was used as a positive control in this test. 10 types of lactic acid bacteria having the intestinal adhesion ability were selected. The selected 10 types of lactic acid bacteria were treated in 3T3-L1 cells and intracellular fat was stained with Oil Red O reagent to measure fat accumulation. As a result, in the group treated with lactic acid bacteria OS-15, 48, 102 and 120, fat accumulation was significantly decreased compared with the non-treated group. And the strain OS-48 showed the greatest inhibitory effect on fat accumulation. The lactic acid bacterium OS-48 was identified as Lactiplantibacillus plantarum by analysis based on 16S rRNA sequencing and was named Lactiplantibacillus plantarum OS-48. Therefore, based on the results of this study, L. plantarum OS-48 is expected to be very useful as probiotics with anti-obesity effect in the food industry such as fermented food and functional food.

본 연구는 생후 12개월령의 염소를 사용하여 앞다리, 뒷 다리, 등심 및 갈비 부위로 분할하여 in vitro 소화실험을 통해 부위별 단백질 가수분해도 및 아미노산 조성을 조사 하였다. 이 때, 소고기 및 돼지고기의 분할육을 이용하여 염소고기와 비교, 분석하였다. 염소고기 분할육 중 뒷다리 (8.32%) 및 갈비(8.32%)가 가장 높게 단백질 가수분해도가 나타났으며, 염소고기의 갈비 부위는 갈비 분할육 중 가장 높은 단백질 가수분해율을 보였던 돼지고기(8.57%)와 유의 차가 없었다 (P>0.05). In vitro 소화 전에는 염소고기 분할 육 중 등심에서 글리신(11.03%)이, 앞다리에서 글루타민 (53.44%)이 다른 고기 종류 및 분할육들에 비해 유의적으 로 높은 비율로 포함된 것이 확인되었다(P<0.05). In vitro 소화 후에는 염소고기 갈비 부위에서 라이신(17.54%)이 가 장 높은 비율로 포함된 것으로 확인되었으며, 소 갈비 부 위보다 유의적으로 높았다(P<0.05). 본 연구는 염소고기 분 할육의 단백질 가수분해도 및 아미노산 조성을 제공하며 단백질 소화양상 및 생체 이용률을 평가하기 위한 기초 자 료로써 활용되어질 수 있을 것으로 사료된다.

본 연구의 목표는 S. boulardii 유래 세포벽 추출물 (CWSB)의 첨가가 반추위 발효에 미치는 영향을 평가하는 것이다. 본 연구의 in vitro 발효 실험은 CWSB의 첨가용 량이 다른 2개의 실험으로 구성되었다. 실험 1에서는 대 조구(CON, 시험사료)와 CWSB0.05(시험사료+CWSB 0.5g/kg 첨가), CWSB0.075(시험사료+CWSB 0.75g/kg 첨 가)의 처리구로 구성되었고, 실험 2의 처리구는 대조구 (CON, 시험사료)와 CWSB0.5(시험사료+ CWSB 5g/kg 첨 가), CWSB1.0(시험사료+CWSB 10g/kg 첨가)의 처리구로 구성되었다. In vitro 배양 이후 3, 6, 12, 24, 48h 동안 발 생한 가스 발생량 및 48h 배양 이후 발효 성상을 분석하 였다. 실험 1에서는 CWSB0.05 처리구에서 유의적으로 높 았던 Vmax(p<0.01)를 제외한 모든 반추위 발효성상에서 처 리구간 유의적 차이가 관찰되지 않았고, 이에 따라 사료 내 0.5%, 1%의 고용량 첨가 수준을 설정한 실험 2를 수행 하였다. 실험 2에서의 CWSB1.0 처리구는 대조구에 비해 총 VFA 발생량이 유의적으로 높았으나 전체 배양시간 내 가스 발생량 및 건물 소화율, 섬유소 소화율은 대조구와 유의적인 차이를 나타내지 않았다. 본 연구 결과를 종합 해 볼 때, CWSB의 첨가수준은 0.5% 이상에서 반추위 발 효 개선효과를 나타낼 것으로 추정된다.

반추위 미생물체 단백질의 아미노산 조성이 한우의 에 너지원 사료에 따라 차이가 있는지를 조사하고자 본 연구 가 수행되었다. 본 실험은 국내 한우 비육우 사육에 주로 이용되는 에너지원 사료인 옥수수(T1), 생미강(T2), 소맥 (T3), 소맥피(T4)를 이용하여 in vitro 반추위 발효실험을 진행하여 발효 24시간 후 배양액의 발효 성상, 미생물체 단백질 합성량(MPS), 미생물의 아미노산 조성 및 미생물 균총 변화를 분석하였다. 배양액의 pH는 T4가 6.02로 다 른 처리구들에 비해서 가장 높았으며, T1은 4.84로 가장 낮았다(p<0.05). Total VFA 농도는 T2가 96.80mM로 다른 처리구들에 비해서 가장 높았고, T1이 53.35mM로 가장 낮았다(p<0.05). 미생물체 단백질 합성량은 T3가 222.50g/L로 다른 처리구들에 비해서 높았으며(p<0.05), T1과 T4는 유의적인 차이는 없었지만 T1이 가장 낮았다. DGGE band 분석결과 모든 처리구는 57.5% 이하의 유사도를 나타내었으며, 급여되는 사료에 따라서 미생물 군집 이 다르게 나타났다. 미생물 내 총 아미노산 함량은 T3와 T4가 다른 처리구보다 높았다(p<0.05). 미생물의 아미노산 조성 중 대부분의 필수 아미노산은 T4가 다른 처리구들에 비해서 낮았다. 하지만 methionine은 다른 필수 아미노산 과 다르게 T4에서 가장 높았다(p<0.05). 곡류 사료 위주의 급여 시 결핍되기 쉬운 lysine은 모든 처리구에서 미생물 의 아미노산 조성이 8.05~8.98%의 범위에 있었다. 따라서, 본 연구에서 반추 가축 급여 에너지 사료원에 따른 반추 위 내 미생물 발효특성과 미생물 군집의 차이가 미생물의 아미노산 조성을 변화시킨다는 것을 확인하였다.

Ulva compressa Linnaeus (UCL) is a green algae seaweed that performs photosynthesis and is used as a food material in some Asian regions including Korea. It is known to be the dominant species in copper ion-contaminated seas, and many studies on copper ion resistant mechanisms have been reported. UCL is known to have an excellent antioxidant effect, but limited information is available regarding its other physiological activities. In this study, we investigated the anticancer activity of 30% prethanol extracts of Ulva compressa Linnaeus (30% PeUCL) and the underlying mechanisms of its activity on human FaDu hypopharyngeal squamous carcinoma cells. The 30% PeUCL extracts suppressed FaDu cell viability without affecting normal cells (L929), as determined by MTT and viability assays. Furthermore, the 30% PeUCL extracts induced apoptosis, as determined by DAPI staining. The 30% PeUCL extracts inhibited colony formation effectively as well as wound-healing of FaDu cells, even at noncytotoxic concentrations. In addition, 30% PeUCL extracts induced apoptosis significantly through proteolytic cleavage of caspase-3, -7, and -9, and poly (ADP-ribose) polymerase, and by downregulation of Bcl-2 and upregulation of Bax in FaDu cells, as determined by Western blot analysis. Collectively, these results suggest that the inhibitory effect of 30% PeUCL extracts on the growth of oral cancer cells, colony formation and wound-healing may be mediated by caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, 30% PeUCL extracts can be administered as a natural chemotherapeutic drug for the treatment of human oral cancers.

Currently, there is no treatment to reverse or cure heart failure caused by ischemic heart disease and myocardial infarction despite the remarkable advances in modern medicine. In addition, there is a lack of evidence regarding the existence of stem cells involved in the proliferation and regeneration of cardiomyocytes in adult hearts. As an alternative solution to overcome this problem, protocols for differentiating human pluripotent stem cell (hPSC) into cardiomyocyte have been established, which further led to the development of cell therapy in major leading countries in this field. Recently, clinical studies have confirmed the safety of hPSC-derived cardiac progenitor cells (CPCs). Although several institutions have shown progress in their research on cell therapy using hPSC-derived cardiomyocytes, the functions of cardiomyocytes used for transplantation remain to be those of immature cardiomyocytes, which poses a risk of graft-induced arrhythmias in the early stage of transplantation. Over the last decade, research aimed at achieving maturation of immature cardiomyocytes, showing same characteristics as those of mature cardiomyocytes, has been actively conducted using various approaches at leading research institutes worldwide. However, challenges remain in technological development for effective generation of mature cardiomyocytes with the same properties as those present in the adult hearts. Therefore, in this review, we provide an overview of the technological development status for maturation methods of hPSC-derived cardiomyocytes and present a direction for future development of maturation techniques.

The present study was conducted to examine the effect of soybean silage as a crude protein supplement for corn silage in the diet of Hanwoo steers. The first experiment was conducted to evaluate the effect of replacing corn silage with soybean silage at different levels on rumen fermentation characteristics in vitro. Commercially-purchased corn silage was replaced with 0, 4, 8, or 12% of soybean silage. Half gram of the substrate was added to 50 mL of buffer and rumen fluid from Hanwoo cows, and then incubated at 39°C for 0, 3, 6, 12, 24, and 48 h. At 24 h, the pH of the control (corn silage only) was lower (p<0.05) than that of soybeansupplemented silages, and the pH numerically increased along with increasing proportions of soybean silage. Other rumen parameters, including gas production, ammonia nitrogen, and total volatile fatty acids, were variable. However, they tended to increase with increasing proportions of soybean silage. In the second experiment, 60 Hanwoo steers were allocated to one of three dietary treatments, namely, CON (concentrate with Italian ryegrass), CS (concentrate with corn silage), CS4% (concentrate with corn silage and 4% of soybean silage). Animals were offered experimental diets for 110 days during the growing period and then finished with typified beef diets that were commercially available to evaluate the effect of soybean silage on animal performance and meat quality. With the soybean silage, the weight gain and feed efficiency of the animal were more significant than those of the other treatments during the growing period (p<0.05). However, the dietary treatments had little effect on meat quality except for meat color. In conclusion, corn silage mixed with soybean silage even at a lower level provided a greater ruminal environment and animal performances, particularly with increased carcass weight and feed efficiency during growing period.

인삼은 다양한 국가에서 과거로부터 민간의약적으로 사용해 온 유용한 한약자원으로, 동북아시아와 북아메리카에서 서식하고 있다. 인삼의 학명은 Panax ginseng C. A. Meyer로 Araliaceae에 속하고 약용 부위는 곁뿌리를 제거한 뿌리 부분이다. 본 연구에서는 전세계에 분포하는 인삼의 종류와 명칭을 정리하고, 동북아시아 공정서 내 인삼의 기원들을 알아보았다. 다양한 인삼종의 전초의 형태를 비교하였고, 여러 문헌 정보를 바탕으로 각국 인삼종의 약리학적 효능을 정리하였다. 현재 한국에서 재배되고 있는 고려인삼의 품종은 약 15종이며, 이들 중 재래종(자경종), 천풍, 연풍, 금풍, 산양삼의 형태학적 특징을 비교하였다. 한약재 활용 가능한 인삼을 수확하기 위해서는 오랜 기간 재배해야 하며, 그 조건도 까다롭다. 약리학적으 로 가치가 있는 인삼종의 시장 수요를 충족하기 위한 대량증식 방법으로 기내조직배양이 활용될 수 있다. 따라서, 각 인삼종의 종자를 수집하여 크기를 측정하고 형태를 비교하였고, 종자를 횡단면으로 절단해 배의 형태를 관찰했다. 각 인삼종의 종자를 활용하여 기내배양을 진행하였고, MS+GA 1.0mg/L 조건에서 발아율은 미국삼이, 생존율은 금풍이 가장 우수한 것으로 확인 되었다. 인삼의 한 종류인 산양삼은 자연에서 성장 속도가 느리고, 재배조건이 까다롭다는 특징을 가지고 있다. 따라서, 산양삼을 수경 재배해, 해당 인삼종에서 발견되는 다양한 미생물종을 동정하였다. 기내배양을 통한 4종의 인삼종 종자 발아 실험에서도 다양한 미생물종이 확인되었고, rDNA 염기서열 분석을 통해 동정하였다. 본 연구는 한국한의학 연구원이 제공하는 전통의학정보포탈(KIOM Oasis Portal)과 농업진흥청 원예특작과학원의 인삼특작부에서 각 인삼종의 종자를 분양받아 진행되었 다. 해당 논문은 기내배양을 통한 인삼의 대량생산 체계 구축 연구의 초석이 될 것으로 사료된다.

Bulbophyllum auricomum Lindl. is a rare orchid and has flowers with an attractive fragrance. The present study investigated the tissue culture method for micropropagation. Capsules derived from artificial self-pollination were obtained for the best seed germination in MS basal medium. Plant growth regulators (1.0 mg·L-1 of BAP and 2.0 mg·L-1 of NAA) were affected by callus induction from subcultured pseudobulb explants. For the callus subculture, different natural plant extracts were tested in 11 treatment media. Among them, MS medium with 150 mL·L-1 of coconut water was generally effective in fresh weight (1.75 ± 0.08) and (3.01 ± 0.20) of callus proliferation and PLBs induction at 1 and 2 months, respectively, followed by an MS combination of 30 g·L-1 of banana and 20 g·L-1 of potato extract. The results of a comparative study of different MS mediums containing plant growth regulators with a natural extract combination and MS medium supplemented with natural extract only showed that MS medium supplemented with a combination of natural extracts (150 mL·L-1 of coconut water) and plant growth regulators (2.0 mg·L-1 of BAP and 1.0 mg·L-1 of NAA) obtained the highest shoot regeneration (3.37 ± 0.17) and (6.41 ± 0.68) after 1 month and 2 months of culturing, respectively.