Piperlongumine (PL) is a natural product found in long pepper (Piper longum ). The pharmacological effects of PL are well known, and it has been used for pain, hepatoprotection, and asthma in Oriental medicine. No studies have examined the effects of PL on bone tissue or bone-related diseases, including osteoporosis. The current study investigated for the first time the inhibitory effects of PL on osteoclast differentiation, bone resorption, and osteoclastogenesis-related factors in RAW264.7 macrophages stimulated by the receptor activator for nuclear factor- κB ligand (RANKL). Cytotoxicity was examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and osteoclast differentiation and bone resorption were confirmed by tartrate-resistant acid phosphatase (TRAP) staining and pit formation analysis. Osteoclast differentiation factors were confirmed by western blotting. PL exhibited toxicity in RAW264.7 macrophages, inhibiting osteoclast formation and bone resorption, in addition to inhibiting the expression of osteoclastogenesis-related factors, such as tumor necrosis factor receptor-associated factor 6 (TRAF6), c-Fos, and NFATc1, in RANKL-stimulated RAW264.7 macrophages. These findings suggest that PL is suitable for the treatment of osteoporosis, and it serves as a potential therapeutic agent for various bone diseases.

In general, the shape of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage is important roles on meiotic maturation of porcine oocyte during in vitro maturation (IVM). Then, mitochondria produce reactive oxygen species (ROS) such as superoxide from electron transport system during oocyte maturation. ROS levels on oocytes are regulated by various antioxidant enzymes in cumulus cells (CCs). However, the effect of mitochondria derived superoxide production from CCs during IVM of porcine oocyte has not been reported. Firstly, we divided groups according to large number of CCs (Grade 1: G1) and small number of CCs (Grade 2: G2). Then, we counted cumulus cells of G1 and G2 oocyte by using haemocytometer. The oocyte maturation rate was significant decreased (p < 0.05) in G2 oocytes than that of G1 oocytes. We measured mitochondria derived superoxide in G1 and G2 COCs by using Mito-SOX staining. Mitochondrial superoxide was higher in G2 COCs than G1 COCs. Then, the mRNA expression levels of antioxidant enzymes (SOD1, SOD2 and PRDX3) in G2 COCs were decreased compared to G1 COCs. To reduce mitochondria derived superoxide, we used Mito-TEMPO as mitochondrial superoxide scavenger. Oocyte maturation rates in both G1 and G2 groups treated with Mito-TEMPO were increased than that of non-treated groups. Mitochondrial superoxide was lower in G1 and G2 groups treated with Mito-TEMPO than that of non-treatment groups. The mRNA expression levels of antioxidant enzymes in G1 and G2 COCs treated with Mito-TEMPO were increased compared to non-treated groups. Based on these findings, we suggest that reduction of mitochondria derived superoxide by Mito-TEMPO assists maturation competence in porcine oocytes.

본 연구의 목적은 다층자료 매개효과의 분석 방법을 제안하는 것이다. 연구내용은 다층자료 매개효과의 통계방법 탐색, 분석절차 제안, 그리고 분석의 실례 제시 등 세 가지다. 첫째, MLM (multilevel modeling)과 MSEM (multilevel structural equation modeling) 중에서 어떤 방법이 다층자료의 매개효과 분석에 유용한지 탐색하였다. MSEM은 MLM 의 약점을 극복한 것으로서 유용한 다층 매개효과 분석방법이었다. 둘째, 다층자료 매개효과의 분석절차를 연구모형설 정, 전제조건 검토, 모형검증, 계수검증의 4단계로 전개하였다. 셋째, 매개효과 분석의 실례에 사용된 자료는 2,695명의 초중등 학생과 88명의 학급교사로 구성되었다. 분석 실례로 2층→2층→1층과 1층→1층→2층 두 가지를 제시하였다. 2층→2층→1층과 1층→1층→2층 모형은 완전매개모형이 지지되었지만, 2층→2층→1층 모형의 매개효과 계수만 95% 신뢰구간에서 유의하였다. 분석 실례에 사용된 Mplus 프로그램은 부록에 제시하였다. 연구결과를 기초로 본 연구의 의의와 제한점, 그리고 후속연구의 방향이 논의되었다.

동절기 산취(Chamecereus silvestrii) ‘희망’의 주·야 온도를 달리하며 처리한 결과 초장은 주간온도 20oC와 야간온도 15oC에서 가장 크게 나타났다. 구경은 주간온도 25oC에서 구직경과 구고의 생육이 우수하였고, 주간온도 25oC에서는 모든 야간온도에서 구직경 생육이 양호하였으며 구고는 주간온도 25oC에서는 20oC에 비해 야간온도가 25oC로 높을수록 생육이 좋았다. 주·야간온도 조합의 합산온도가 높을수록 구직경과 구고신장이 증가하는 경향을 나타났다. 색도에서 Hunter ‘a’ 값은 야간온도가 높을수록 낮아졌으나 ‘b’ 값은 높아져 야간온도가 높으면 탁도가 증가하였다. 또 한 구직경과 구고변화의 증가가 다시 시작되는 기점은 정 식 후 15일로 나타났다.