말뼈추출물은 다양한 골질환의 예방과 치료에 탁월한 효능이 있다고 이전에 보고되었다. 하지만 말뼈 추출물의 다른 약리학적 효능에 대해서는 아직 자세히 밝혀지지 않고있다. 본 연구에서는 말뼈추출물이 중요한 항산화 인자인 hemeoxygenase-1(HO-1)의 발현을 상승시킬 수 있는지, 만약 발현이 증가한다 면 HO-1의 상향 조절이 대식세포에서 항염증 효과를 매개할 수 있는지에 관하여 조사하였다. 이를 위 해서 nitric oxide(NO) 농도측정, 세포 생존능 측정, DPPH 라디칼 소거능 검사를 시행하였다. 또한 염 증성 사이토카인 유전자 발현과 단백질 발현을 측정하기 위해 real time PCR과 Western blotting을 시행하였다. 말뼈추출물은 lipopolysaccharide(LPS, 0.1μg/ml)로 자극한 대식세포주인 RAW264.7 세포 에서 어떠한 세포독성 없이 NO의 생성을 유의성 있게 억제하였으며 inducible nitric oxide(iNOS)와 cyclooxygenase 2(COX-2)의 발현을 억제하였다. 뿐만 아니라 말뼈 추출물은 염증성 사이토카인인 tumor necrosis factor(TNF)-α와 interleukin(IL)-1β의 발현을 억제하였으며 ERK, JNK 및 p-38 MAPK의 단백질 인산화를 억제하였다. 그리고 말뼈추출물은 HO-1과 NF-E2-related factor-2 (Nrf-2) 의 발현을 증가시켰고 이것은 말뼈추출물이 가지고 있는 항 염증효과를 매개할 수 있는 것으 로 보인다. 즉, 말뼈추출물이 HO-1의 발현을 상향 조절한 반면 ERK1/2의 신호전달 경로에 손상을 주 는 것으로 확인되었으며 이러한 말뼈추출물의 효과가 최종적으로 세포손상과 세포의 과산화 자극으로부 터 세포를 보호 할 수 있는 것으로 사료된다.

대반하[大半夏: Pinellia tripartita(Blume) Schott]는 천남성과에 속하는 다년생 초본식물로써 중국, 일본, 우리나라를 포함한 아시아 전역에 분포하며 꽃과 전초가 아름다워 최근 원예용으로도 많이 이용 되고 있으나 체계적인 대량번식 체계가 구축되어 있지 않다. 본 연구에서는 조직배양을 통해 증식된 대 반하 기내 식물의 토양조성별 괴경 생육을 조사하여 비교하였다. 기내 증식된 대반하 괴경의 크기에 따 라 직경이 1cm 이상인 경우 TypeⅠ, 1cm 이하를 TypeⅡ로 나누어 6종의 조합상토에 파종하여 괴경의 비대와 생육을 조사하였다. 괴경의 비대와 전초의 생육변화를 확인하기 위하여 초장, 잎 수, 마른 잎 수, 괴경 수, 괴경 크기, 생체중 및 건물중을 8주간 변화를 측정한 결과, Type I와 II 모두 코이어 68.0%, 피트모스 14.7%, 펄라이트 3.0%, 버미큘라이트 7.0% 및 제오라이트 7.0%로 구성된 조합상토 B에서 가장 우수한 생육을 보였다. 특히 TypeⅠ은 조합상토 B에서 괴경 크기와 생체중이 각각 45%와 101%를 보였으며, 이는 생육이 가장 좋지 못했던 처리구인 조합상토 E(코이어 14.3%, 피트모스 14.3%, 펄라이트 42.9%, 버미큘라이트 14.3% 및 제오라이트 14.3%)와 약 2배의 차이를 보였다. 이러한 연구 결과는 종자번식이 어려운 대반하의 기내 대량증식을 통한 육묘에 있어 안정적인 토양 순화와 대규모 배양묘 생산을 위한 중요한 기초자료로 활용할 수 있을 것으로 사료된다.

Nanoparticles are widely used in various fields such as electronics, medicines and getting focus on the application in food industry for developing intelligent delivery system with bioactive ingredients or functional nutrients. Basic study on possible toxicological effect of food applicable nanoparticles is required for a practical application in food industry. In this study, size-controlled bovine serum albumin (BSA) nanoparticles were prepared by a desolvation method and their cytotoxicity was investigated. BSA nanoparticles were prepared with mean diameters as 115, 137, 159, and 299 nm, then cytotoxicity was evaluated with RAW 264.7 macrophages as in vitro model. Cell viabilities were significantly affected as increasing nanoparticle concentration. Smaller the sizes of nanoparticles, LD50 values were significantly reduced. LD50 values of BSA nanoparticles were 50, 65, 126, and 170 μg/ml, respectively. Nanoparticle was supposed to induce the apoptosis of RAW 264.7 marcrophages and underlying mechanism will be investigated in future. These findings will be used as valuable basement for nanofood development with BSA nanoparticles.

As one of the staple crops, rice has been widely applied to value-added products, giving the food industry new avenues of use. Although the quality attributes of various rice products have been reported, there is a lack of detailed information on the rheological behaviors of rice products during digestion that are related to their bioaccessibility in the human body. In this study, three rice varieties with different amylose contents were utilized to produce flours and extruded noodles. In-vitro methods simulating starch digestion processes were then established to monitor their oral-gastric-intestinal rheological behaviors. The rice flour with high amylose content exhibited lower values of water absorption index/swelling power and higher pasting parameters that were in good agreement with the Mixolab thermo-mechanical results. The extruded rice noodles showed lower cooking loss and higher hardness with increasing levels of amylose. When the in-vitro viscosities of rice flours and noodles were measured using a rotational rheometer with the custom-made starch cell, their viscosities had a tendency to decrease as the in-vitro digestion progressed. Specifically, the rice samples with high amylose content exhibited higher viscosity than those with low amylose content under the simulated oral, gastric, and intestinal conditions. Hence, this study was conducted to investigate the physicochemical and in-vitro rheological properties of rice flours and extruded noodles with different amylose content. The results provided a promising opportunity for the food industry to study in-vitro digestion of rice-based products with the advantages of being more rapid and less expensive.

The objective of this study was to determine the effect of bacterial inoculation (Lactobacillus plantarum or combo inoculant mixed with Lactobacillus plantarum and Lactobacillus buchneri) and addition of fibrolytic enzyme on chemical compositions and fermentation characteristics of whole crop barley (WCB) and triticale (TRT) silage, their ruminal in vitro fermentation, and digestibility. In TRT silage, enzyme addition significantly (p<0.01) decreased NDF content compared to no enzyme addition treatment. Organic acids such as lactate and acetate contents in WCB and TRT silages were significantly (p<0.01) higher compared to those in the control. Particularly, lactate content was the highest in L. plantarum treatment. Fibrolytic enzyme treatment on both silages had relatively higher lactic acid bacteria content, while mold content was lower in both treatments compared to that in the control. In vitro dry matter digestibility was generally improved in WCB silages. It was higher (p<0.01) in TRT with mixed treatment of L. plantarum, L. buchneri, and enzyme compared to others. In vitro ruminal acetate production was relatively higher in treatments with both enzyme and inoculant additions compared to that in the control. Therefore, the quality of silage and rumen fermentation could be improved by inoculants (L. plantarum and L. buchneri) regardless whether whole crop barley (WCB) or triticale (TRT) silage was used. Although it was found that fibrolytic enzyme addition to both silages had various quality and rumen fermentation values, further study is needed

Cryopreservation has been applied successfully in many mammalian species. Nevertheless, pig embryos, because of their greater susceptibility to cryoinjuries, have shown a reduced developmental competence. The aim of this study was to evaluate the survival status of vitrified-warmed porcine embryos. Forced blastocoele collapse (FBC) and non-FBC blastocysts are vitrified and concomitantly cultured in culture media which were supplemented with/without fetal bovine serum (FBS). Porcine vitrified-warmed embryos were examined in four different methods: group A, non- FBC without FBS; group B, non-FBC with FBS; group C, FBC without FBS; group D, FBC with FBS. After culture, differences in survival rates of blastocysts derived from vitrified-warmed porcine embryos were found in group A∼D (39.5 (A) vs 52.5 (B) and 54.8 (C) vs 66.7% (D), respectively, p<0.05). Reactive oxygen species (ROS) level of survived blastocysts was lower in group D than that of another groups (p<0.05). Moreover, total cell number of survived blastocysts was higher in group D than that of other groups (p<0.05). Otherwise, group D showed significantly lower number of apoptotic cells than other groups (2.0±1.5 vs 3.2±2.1, 2.8±1.9, and 2.7±1.6, respectively, p<0.05). Taken together, these results showed that FBS/FBC improves the developmental competence of vitrified porcine embryos by modulating intracellular levels of ROS and the apoptotic index during the vitrification/warming procedure. Therefore, we suggest that FBS and FBC are effective treatment techniques during the vitrification/warming procedures of porcine blastocysts.

Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are oocyte-specific growth factors that regulate many critical processes involved in early folliculogenesis and oocyte maturation. In this study, effects of GDF9 and BMP15 treatment during in vitro maturation of porcine oocytes upon development after parthenogenetic activation were investigated. Neither GDF, BMP15 alone nor in combination affects the number and viability of cumulus cells or the rates of oocyte maturation and blastocyst development. However, the treatment of GDF9 on porcine oocytes increased the number of trophectodermal (TE) cells of blastocysts derived from activated oocytes (P<0.05). The treatment of BMP15 increased the cell numbers of both inner cell mass (ICM) and TE cells (P<0.05). The treatment with the combination of GDF9 and BMP15 further increased the numbers of ICM and TE cells, compared with GDF9 or BMP15 treatment alone (P<0.05). In conclusion, the treatment of GDF9 or BMP15 (or both) enhanced the quality of blastocysts via the increased number of ICM and/or TE cells.

Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat β-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

In this study, to improve the in vitro development of various cells including cloned embryos, the effects that isoproterenol and melatonin have on in vitro development of porcine parthenogenetic oocytes were investigated. Parthenogenetic activation was induced with electrical stimulation, BSA and 6-DMAP treatment. 10-7 M of melatonin and isoproterenol (10-10, 10-12 and 10-14 M) were supplemented for in vitro maturation (IVM) and in vitro culture (IVC) medium, with different concentrations. When isoproterenol and melatonin were supplemented in IVM medium with different concentrations, there was no significant (P<0.05) difference of maturation rate in the treatment groups as well as in that of only melatonin. As isoproterenol and melatonin were supplemented in IVM medium with different concentrations, blastocyst rates of isoproterenol 10-12 M treatment group (37.1%) were significantly (P<0.05) higher than control group (26.0%). Isoproterenol and melatonin were supplemented in IVC medium with different concentrations, then the cleavage rate of 10-12 M isoproterenol treatment group (82.2%) was significantly (P<0.05) higher than the group that melatonin was only supplemented (70.9%). There was no difference of blastocyst rate between the treatment groups. When isoproterenol and melatonin were supplemented for IVM+IVC medium with different concentrations, the cleavage rate of 10-12 M isoproterenol treatment group (92.5%) was significantly (P<0.05) higher than the control group (82.8%) and the group that melatonin was only treated (81.6%). The blastocyst rate of 10-12 M as 45.6% was significantly (P<0.05) higher than control group (25.2%) and melatonin treatment group (31.2%). The cell number of blastocyst in 10-12 M isoproterenol treatment group 35.5±3.4 was significantly (P<0.05) highest. The results of this study showed that the development rate of IVC when both isoproterenol and melatonin were supplemented was higher than when melatonin was only supplemented. Therefore, it is concluded that isoproterenol is rather effective in the activation of melatonin. 10-7 M melatonin and 10-12 M isoproterenol were considered suitable concentration.

This study investigated the antibacterial effects of Galla rhois extract (GRE) against Campylobacter jejuni and Campylobacter coli. The minimum inhibitory concentrations (MICs) of GRE against C. jejuni and C. coli were 0.28 and 0.55 mg/mL, respectively, and the corresponding minimum bactericidal concentrations (MBCs) were 4.4 and 5.5 mg/mL. C. jejuni treated with the MIC, MBC or 2×MBC of GRE showed significant inhibition of growth compared with that of the control group during the incubation period, and no viable bacteria were detected at 24 h after incubation. C. coli treated with MIC, MBC or 2×MBC of GRE also showed inhibition of growth compared with that of the control group during the incubation period, and in the C. coli cultures treated with MBC and 2×MBC of GRE, no viable bacteria were detected at 24 h after incubation. In conclusion, GRE is a candidate antibacterial agent against C. jejuni and C. coli, and may have applications for the control of Campylobacter infection in poultry.

Allergic diseases have rapidly increased in recent years. Mast cells play a critical role in IgE-mediated allergy responses and, therefore, closely associated with rhinitis, asthma, and atopic dermatitis. We studied antiallergic effect of Penthorum chinense extract (PCE) in vitro and in vivo. PCE inhibited the degranulation of mast cells by antigen stimulation and its effect was dose-dependent and reversible in mast cells. PCE also suppressed the production of inflammatory cytokines such as TNF-α and IL-4 by antigen in mast cells. Mechanistically, PCE inhibited the activation of Syk/LAT pathway which is a key signaling pathway for the activation of mast cells by antigen. Notably, PCE suppressed IgE-mediated allergic responses by antigen in mice. These results strongly suggest that PCE is a potential candidate for anti-allergic treatment.

Atopic dermatitis (AD) is usually caused by foods such as wheat, egg, milk, and peanuts, leading to common health problems in early childhood with complications like urtication. The aim of this study was to evaluate ethanol extracts of rice and rice snacks concentrated until the ethanol was completely eliminated and hot-air dried. In vitro analyses were carried out using murine macrophage RAW 264.7 cells. We measured cytotoxicity, nitric oxide (NO) production, and inflammatory cytokine level. The NO level of the cells exposed to lipopolysaccharide (LPS) was significantly reduced by rice and rice snack extracts. TNF-α level decreased in contrast to the LPS group, although a significant difference was not observed. On the other hand, IL-6 significantly decreased in both rice and rice snack extracts in a dose-dependent manner. The results of the present study suggest that rice and rice snack decreased NO and inflammatory cytokine levels. Therefore, rice could be useful as a raw material for relieving child atopic dermatitis caused by snacks made from wheat.

The objectives of the present study were to select an effective basic medium including its hormone and protein supplementation for IVM of oocytes of indigenous zebu cows. The ovaries of cows were collected from slaughter house and the follicular fluid was aspirated from 2 to 8 mm diameter follicles. The COCs with more than 3 cumulus cell layers and homogenous cytoplasm were selected for maturation. The oocytes were matured in media for 24 hrs at 39℃ with 5% CO2 in humidified air. The maturation of oocytes was evaluated by examining the presence of first polar body under microscope. An efficient basic medium was determined after culturing COCs in either TCM 199 or SOF medium in Experiment 1. An efficient hormone supplementation was determined after culturing COCs in either FSH or gonadotrophin supplemented TCM 199 in Experiment 2. An efficient protein supplementation was determined after culturing COCs in either FBS or Oestrous cow serum (OCS) supplemented TCM 199 in Experiment 3. The oocyte recovery rate per ovary was 3.35. The overall rate of IVM was 74.6%. The maturation rate was 75.5±3.9 and 62.2±20.2% in TCM and SOF medium, respectively (P>0.05). The maturation rate of oocytes was significantly higher (76.6±13.2%) in FSH supplemented medium than gonadotrphin supplemented counterpart (69.7±10.8%) (P<0.05). The maturation rates of oocytes were 81.7±12.9 and 85.7±12.7% in medium supplemented with FBS and OCS, respectively (P>0.05). In conclusions, both TCM 199 and SOF supplemented with either FBS or OCS, and FSH may be used as medium for IVM of indigenous zebu oocytes in Bangladesh.

Gangliosides exist in glycosphingolipid-enriched domains on the cell membrane and regulate various functions such as adhesion, differentiation, and receptor signaling. Ganglioside GM3 by ST3GAL5 enzyme provides an essential function in the biosynthesis of more complex ganglio-series gangliosides. However, the role of gangliosides GM3 in porcine oocytes during in vitro maturation and early embryo development stage has not yet understood clear. Therefore, we examined ganglioside GM3 expression patterns under apoptosis stress during maturation and preimplantation development of porcine oocytes and embryos. First, porcine oocytes cultured in the NCSU-23 medium for 44 h after H2O2 treated groups (0.01, 0.1, 1 mM). After completion of meiotic maturation, the proportion MII (44 h) was significantly different among control and the H2O2 treated groups (76.8±0.3 vs 69.1±0.4; 0.01 mM, 55.7±1.0; 0.1 mM, 38.2±1.6%; 1 mM, P<0.05). The expressions of ST3GAL5 in H2O2 treated groups were gradually decreased compared with control group. Next, changes of ST3GAL5 expression patterns were detected by using immunofluorescene (IF) staining during preimplantation development until blastocyst. As a result, we confirmed that the expressions of ST3GAL5 in cleaving embryos were gradually decreased (P<0.05) according to the early embryo development progress. Based on these results, we suggest that the ganglioside GM3 was used to the marker as pro-apoptotic factor in porcine oocyte of maturation and early embryo production in vitro, respectively. Furthermore, our findings will be helpful for better understanding the basic mechanism of gangliosides GM3 regulating in oocyte maturation and early embryonic development of porcine in vitro.

In vitro culture of murine embryos is an important step for in vitro production systems including in vitro fertilization and generations of genetically engineered mice. M16 is widely used commercialized culture media for the murine embryos. Compared to other media such as potassium simplex optimization medium, commercial M16 (Sigma) media lacks of amino acid, glutamine and antibiotics. In the present study, we optimized M16 based embryo culture system using commercialized antibiotics-glutamine or amino acids supplements. In vivo derived murine zygote were M16 media were supplemented with commercial Penicillin-Streptomycin-Glutamine solution (PSG; Gibco) or MEM Non- Essential Amino Acids solution (NEAA; Gibco) as experimental design. Addition of PSG did not improved cleavage and blastocyst rates. On the other hand, cleavage rate is not different between control and NEAA treated group, however, blastocyst formation is significantly (P<0.05) improved in NEAA treated group. Developmental competence between PSG and NEAA treated groups were also compared. Between two groups, cleavage rate was similar. However, blastocyst formation rate is significantly improved in NEAA treated group. Taken together, beneficial effect of NEAA on murine embryos development was confirmed. Effect of antibiotics and glutamine addition to M16 media is still not clear in the study.

The present in vitro study was conducted to examine the effect of buffer solubility of eight protein feeds (coconut meal, distillers grain, sesame meal, perilla meal, soy source cake, rape seed meal, soybean meal and lupine) on the fermentation characteristics, degradability of dry matter (DM) and crude protein (CP), and methane (CH4) production by rumen microbes. Buffer extraction increased pH (P<0.05 ~ p<0.001) of the culture solution but tended to lower ammonia- N concentration for all protein feeds. Total volatile fatty acids(VFAs) and each VFAs concentrations in all incubation was decreased by buffer extraction (P<0.01 ~ P<0.001). Also, molar proportion of acetate in 1h (P<0.001), 3h (P<0.01) and 12h (P<0.05) incubations and molar proportion of propionate in 1h (P<0.001), 3h (P<0.01), 6h (P<0.05) and 12h (P<0.05) were decreased by buffer extraction. But molar proportion of butyrate in 1h (P<0.001), 3h (P<0.01) and 6h (P<0.05) were increased by buffer extraction. The in vitro effective degradability of dry matter (P<0.001) and CP (P<0.001) was decreased by buffer extraction. The methane production (P<0.01~P<0.001) in all incubation was decreased by buffer extraction. The results from in the current study might be useful for diet formulation to improve the feed efficiency of the ruminant animals without massive loss of major nutrients.

This study attempted to find an efficient method for the preparation of high-purity galactooligosaccharides (HP-GOS) using β-galactosidase and yeast fermentation. GOS prepared using Lactozym 3000L showed the greatest enhancement in total GOS of the six β-galatosidases tested. GOS alone achieved 51% conversion of initial lactose. GOS production was enhanced by fermentation with commercial yeast (Saccharomyces cerevisiae); its concentration reached 71% after 36h fermentation with 8% yeast. Component sugar analysis with HPLC indicated that HP-GOS fermented with S. cerevisiae showed significantly increased levels of 4’/6’-galactosyllactose and total GOS as well as a significantly decreased glucose level. HP-GOS facilitated the growth of Lactobacillus sp. (L. acidophilus and L. casei) and Bifidobacterium sp. (B. longum and B. bifidum). In sum, high-purity GOS has been successfully produced through both an enzymatic process and yeast fermentation. GOS encourages the growth of bacteria such as Lactobacillus and Bifidobacterium that may be beneficial to human gastrointestinal health.