Changes in contents of free sugars, amino acids, and fatty acids of legumes were analyzed for each phase of in vitro digestion. In addition, contents of resistant starch in raw and digested pulses were compared. Soybeans, kidney beans, cowpeas, and chickpeas were analyzed. An in vitro digestion model was used to analyze contents of nutrients using LC-MS and GC-MS. Stachyose in kidneybean, cowpea, and chickpea increased as the digestion phase progressed. In four types of legumes, raffinose slightly decreased or showed no significant difference between the Oral phase and the BBMV phase. Content of glucose, a monosaccharide, increased during the BBMV phase. During the digestion phase, levels of free amino acids and free fatty acids also increased. Content of resistant starch was reduced compared to that in the raw material. It was 0.01g/100 g food in soybean, 1.06 g/100 g food in red kidney bean, 0.77g/ 100g food in cowpea, and 0.76 g/100 g food in chickpea. It was confirmed that nutrients in the in vitro digestion model were liberated at each digestion phase with changes in the content of resistant starch. These results are expected to be used as fundamental data for obtaining bioavailability of nutrients.

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.

In vivo 측정법은 비용이 많이 들고 연구기간이 길며 동물복지 측면에서 여러 문제점이 있다. 이에 대한 대안으로 지난 수십년간 in vitro 소화율 측정법이 개발되어 왔다. 본 총설에서는 대표적인 단위동물로써 사양되고 있는 돼지의 소화기관 특징 및 소화작용에 대한 논하였다. 또한, 최근에 개발된 daisy II incubator 를 이용한 in vitro 방법을 통해 돼지에서 주요 곡류사료로 이용되고 있는 옥수수, 쌀, 소맥 및 대맥의 회장 및 전분 건물소화율을 측정하였다.

Mammalian fetal ovaries contains numerous primordial germ cells, however fewer ones can yield mature oocytes due to apoptosis and follicle atresia. Successful in vitro reconstitution of primordial germ cells has recently had a significant effect in the field of assisted reproductive technologies. However, the regulatory mechanisms underlying oogenesis remain unknown and recapitulation of oogenesis in vitro remains unachieved. Therefore, development of methods for obtaining mature oocytes by culturing the fetal ovaries in vitro could contribute to clarify these mechanisms. We adapt an in vitro system for culturing mouse fetal ovaries that support successful follicle assembly and improve oocyte growth and maturation. Ovarian tissues from 12.5 days postcoitum (dpc) fetal mice were cultured in vitro and the matured oocytes were differentiated from primordial germ cells after a 31 days culture period. Our results demonstrate that mouse fetal germ cells are able to form primordial follicles with artificial ovarian cells, and that oocytes within the growing follicles are able to mature normally in vitro. Taken together, this in vitro culture system is expected to aid in the development of new strategies to identify the reasons behind failure of follicle assembly and offer a platform for innovative research into preservation of female germ cells and conservation of endangered species.

귀리, 콩, 감자, 밀에서 유래한 다양한 식이섬유를 밀가루 대신 10-40%까지 대체함에 따른 수화능, 동적점탄성, 용매흡착능 및 in vitro starch digestion을 비교하였다. 귀리, 콩, 감자, 밀에서 유래한 다양한 식이섬유를 식이섬유의 대체비율이 증가할수록 수분흡착능과 팽윤력은 증가한 반면, 수분용해도는 감소하였다. 밀가루 대신 콩과 감자 식이섬유를 대체함에 따라 동적점탄성은 증가하였으나, 귀리와 밀 식이섬유 대체에서는 오히려 감소하는 경향을 보였다. 식이섬유 대체에 따른 밀가루의 용매흡착능 변화를 분석한 결과, 낮은 대체량의 귀리 식이섬유와 높은 대체량의 콩과 감자 식이섬유가 활용 가능성을 보였다. 원료별 식이섬유 대체 시 밀가루 겔의 전분소화 패턴은 식이섬유 대체비율이 증가함에 따라 glucose 방출은 감소하였으나, 원료별 특성에 따른 유의적인 차이는 없었다. 특히, 밀식이섬유를 제외한 귀리, 콩, 감자 식이섬유는 모두 RDS감소와 RS증가에 따른 pGI 저하를 보여 실제 식품소재로 활용 시 가공적성을 유지할 수 있다면 전분소화지연효과를 기대할 수 있음을 확인할 수 있었다. 한편, 식이섬유의 원료에 따른 식이섬유 조성과 전분소화 관련 특성 간 상관관계를 분석한 결과, 전분소화를 지연시키는 효과는 수용성 식이섬유보다는 불용성과 총식이섬유 함량이 중요한 인자임을 알 수 있었다.

Royal jelly (RJ) is exclusive food that is secreted from the hypopharyngeal and mandibular glands of worker honeybees, and it is well known to be a necessary for the growth of the queen honeybee Although fresh royal jelly have been demonstrated to enhance wound healing, the wound healing effects of water soluble royal jelly (WSRJ) have not been elucidated. We investigated whether WSRJ promotes the migration, attachment, and proliferation of human dermal fibroblasts (HDFs) during in vitro wound healing. HDFs were treated with 1-5ug/ml WSRJ and RJ for up to 24hr following wound formation. Cell migration was assessed by measuring recovery from wound margin, while cell attachment and proliferation were determined by MTT assay. By observing the numbers of cell attached, we confirmed that not only WSRJ but also RJ did not affect on the initial cell adhesion. WSRJ (5 ug/ml) enhanced cell migration rate approximately 84.3% in HDFs at 24hr, whereas RJ (5 ug/ml) increased cell migration rate 71.3% in HDFs at 24hr, which is similar to cell migration rate of WSRJ 1 ug/ml (73.7%). In cell proliferation assays, WSRJ induced an increase in the number of HDFs, compared with control and RJ. In conclusion, WSRJ promotes cell migration with increased cell proliferation in an in vitro wound healing model.

Peri-implantitis (PI) is bacteria-induced inflammatory condition which affects the alveolar bone and soft tissue around implants and may result in the loss of supporting bone. Attenuation of the P. gingivalis lipopolysaccharide (LPS)-induced inflammatory response can be a new therapeutic approach in the treatment of PI. This study was conducted to evaluate the anti-inflammatory effect of 635-nm light-emitting diode (LED) irradiation over MG63 osteoblast-like cell. Scratch was made on MG63 cells with or without LPS, then 635-nm irradiated. The expression of the cyclooxygenase-2 (COX-2) proteins was evaluated with western blot. The production of the prostaglandin E2 (PGE2) and expression of the receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG) was measured with enzyme-linked immunoassay, and the cytokine profile was evaluated with the human inflammation antibody array. Wound closure effect presented in the cells treated with LPS was observed more significantly in the cells with 635-nm irradiation than the cells without irradiation. The 635-nm irradiaiton reduced LPS-induced expression of the COX-2 and production of the PGE2. Also, 635-nm irradiation affect the expression of RANKL, OPG, and proinflammatory cytokines. These results indicate that 635-nm irradiation could reduce the alveolar bone resorption induced by LPS stimulation through the inhibition of COX-2 expression and PGE2 production, the suppression of proinflammatory cytokine, and the modulation of RANKL/OPG balance in MG63 cells.

본 연구에서는 대표적인 퇴행성 신경질환인 알츠하이머성 치매에 대한 마늘 물, 100% 메탄올, 디 클로로메탄 추출물들의 acetylcholinesterase (AChE) 저해 및 신경세포 보호효과를 조사하였다. 마늘 디클로로메탄 추출물은 농도 의존적으로 AChE를 저해하는 것으로 나타났으며, IC50은 36.1 μg/mL로 나타났다. MTT reduction assay를 이용해 amyloid β protein (Aβ) 유도성 신경세포 독성에 대한 신경 세포 보호효과를 측정한 결과, 세 가지 마늘 추출물들은 대부분 40% 미만의 세포생존율을 보였고 이 결과는 Aβ 유도성 신경세포 독성보다 상대적으로 더 높은 세포독성을 보여주었다. LDH assay에 서는 마늘 물 추출물이 37%의 LDH 방출량을 나타내 200 μM의 vitamin C과 유사한 세포막손상 보 호효과를 보였다. 마지막으로 neutral red uptake assay를 실시한 결과, MTT reduction assay와 마찬가 지로 모든 마늘 추출물들에서 세포생존율의 감소를 확인하였으며 특히 디클로로메탄 추출물의 경우 현저하게 낮은 세포생존율을 나타내었다. AChE 저해활성을 갖는 마늘 디클로로메탄 추출물로부터 얻은 column fractionations에 함유된 생리활성물질을 탐색하기 위해 HPLC 분석을 실시하였으며, 마 늘 98:2 fraction의 LC-MS 분석을 통하여 allyl methyl disulfide, diallyl monosulfide, diallyl disulfide로 추정되는 물질군이 확인되었다.

Human embryonic stem (hES) cells can be induced to differentiate into tyrosine hydroxylase expressing (TH+) cells that may serve as an alternative for cell replacement therapy for Parkinson's disease (PD). To examine in vitro differentiation of hES (MB03, registered in NIH) cells into TH+ cells, hES cells were induced to differentiate according to the 4-/4+ protocol using retinoic acid (RA), ascorbic acid (AA), and/or lithium chloride (LiCl) followed by culture in N2 medium for 14 days, during which time the differentiation occurs. Immunocytochemical stainings of the cells revealed that approximately 21.1% of cells treated with RA plus AA expressed TH protein that is higher than the ratio of TH+ cells seen in any other treatment groups (RA, RA+LiCl or RA+AA+LiCl). In order to see the differentiation pattern in vivo and the ability of in vitro differentiation-induced cells in easing symptomatic motor function of PD animal model, cells (2 10 cells/2) undergone 4-/4+ protocol using RA plus AA without any further treatment were transplanted into unilateral striatum of MPTP-lesioned PD animal model (C57BL/6). Following the surgery, motor behavior of the animals was examined by measuring the retention time on an accelerating rotar-rod far next 10 weeks. No significant differences in retention time of the animals were noticed until 2 weeks post-graft; however, it increased markedly at 6 weeks and 10 weeks time point after the surgery. Immunohistochemical studies confirmed that a reasonable number of TH+ cells were found at the graft site as well as other remote sites, showing the migrating nature of embryonic stem cells. These results suggest that in viかo differentiated hES cells relieve symptomatic motor behavior of PD animal model and should be considered as a promising alternative for the treatment of PD.