Search

Purpose: This study evaluated the biological activity of five types of mints (Peppermint, Spearmint, Chocomint, Applemint, Pineapplemint).Methods: Five types of mints were extracted with an ethanol solvent to compare the total polyphenol and total flavonoid content, scavenging activities of DPPH and ABTS radical, SOD-like activity, and inhibitory effects of elastase, tyrosinase, and α-glucosidase.Results: All five types of mint extracts were confirmed to be effective significantly in concentration. The total polyphenol content was found to be effective in the order of peppermint>spearmint>chocomint>pineapplemint>applemint, and the total flavonoid content was found to be effective in the order of peppermint> chocomint>spearmint>pineapplemint>applemint. The DPPH radical scavenging activity was found to be in the order of peppermint>spearmint>chocomint> pineapplemint>applemint, and the ABTS radical scavenging activity was found to be in the order of peppermint>chocomint>spearmint>pineapplemint>applemi nt. The SOD-like activity was found to be in the order of peppermint>chocomint> spearmint> pineapplemint>applemint. The inhibitory effects of elastase, tyrosinase, α-glucosidase were found to be in the order of peppermint>spearmint>chocomint> pineapplemint>applemint.Conclusion: As above results, all five types of mint extracts were verified to be effective significantly in concentration. In particular, peppermint indicated the highest total polyphenol, total flavonoid, DDPH and ABTS radical scavenging activity, and SOD-like activity, while chocomint displayed the highest tyrosine, elastase, and α-glucosidase inhibitory effects.

Purpose: This study aims to confirm the potential of rosemary, parsley, thyme, chive, and dill extracts as natural antimicrobial agents and incense cosmetic materials by evaluating the antioxidation effects and antimicrobial effects on the skin and related microorganisms.Methods: Rosemary, parsley, thyme, chive, and dill were extracted with 70% ethanol and used as a sample. Antioxidant effects were measured at the total polyphenol and flavonoid content, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, 2,2'-azinobis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging effects and superoxide dismutase (SOD)-like activity effect. And for Staphylococcus epidermidis, Propionibacterium acnes, Corynebacterium xerosis, Malassezia furfur, and Trichophyton mentagrophytes related to the skin, the antimicrobial effect was measured using the paper disc method.Results: Among the rosemary, parsley, thyme, chive, and dill extracts, the total polyphenol and flavonoids were found to be high in rosemary and parsley extracts. DPPH and ABS radical scavenging effects and SOD-like activity effects were confirmed to be in the order of rosemary>parsley>thyme> chives>dill. As a result of measuring the antimicrobial effects at a concentration of 5 mg/mL, S. epidermidis was confirmed to be in the order of parsley=rosemary=thyme>chive=dill, P. acnes was confirmed to be in the order of rosemary>parsley>dill>thyme=chive, C. xerosis was confirmed to be in the order of rosemary>parsley>dill>thyme=chives, M. furfur was confirmed to be in the order of rosemary>parsley>dill>chives>thyme, and T. mentagrophytes was confirmed to be in the order of rosemary>parsley>chives>dill>thyme.Conclusion: Rosemary, parsley, thyme, chive, and dill extracts were confirmed to have antioxidant and antimicrobial effects, confirming their potential as natural antimicrobial agents and incense cosmetic materials.

PDF file, 1016KB, Supplementary Figure 1. Immunohistochemical staining shows L1CAM is expressed in the invasive areas of ATC. Cases 1 through 3 have L1CAM expression in the invasive islands (red arrowhead). Cases 7 and 8 express L1CAM in the invasive fronts of ATC (black arrowhead). Supplementary Figure 2. Aberrant L1CAM expression is observed in recurrent ATC. Case 4 has negative L1CAM expression at the PTC tumor (left). Black arrowhead indicates the internal positive control at the peripheral nerve bundles that has normal L1CAM expression. However, the recurred ATC tumor exhibits positive L1CAM expression in Case 4 (right). Supplementary Figure 3. Immunohistochemical staining of cyclin D1 and p53 in ATC (Original magnification: �200). Supplementary Figure 4. L1CAM expression in primary cultured follicular cells (PCFC), primary cultured PTC (PCPTC), PTC cell lines (TPC1 and BCPAP), and ATC cell lines (FRO and 8505C). The ovarian carcinoma cell line SKOV3 is used as a positive control. Thyroglobulin is a marker of thyroid-derived cells.

PDF file, 118KB.

Purpose: Recently, research and development on natural materials are actively carried out. It was conducted to confirm the applicability of Neem and Comfrey extracts as natural preservatives and cosmetic ingredients by investigating their antioxidative, antibacterial, and antifungal activities. Methods: Both Neem and Comfrey were extracted with 70% ethyl alcohol as samples, respectively. Their antibacterial and antifungal activities for antioxidative effect and against skin-related bacteria were confirmed by the Paper Disc method. Results: Both extracts were found to be highly effective in antioxidative effects, but the Comfrey extract showed higher total polyphenols and flavonoid content, stronger scavenging capability of 1,1-Diphenyl-2-picryhydrazyl (DPPH) and 2,2'-azino-bis-3-ehtylbenzothiazoline-6- sulfonic acid (ABTS) radicals as well as more robust superoxide dismutase (SOD) like activities than the Neem extract did. After measuring antibacterial and antifungal effects by the Neem and the Comfrey extracts against Staphylococcus epidermidis (S. epidermidis ), Staphylococcus aureus (S. aureus ), Propionibacterium acnes (P. acnes ), Corynebacterium xerosis (C. xerosis ), Pityrosporum ovale (P. ovale ), Malassezia furfur (M. furfur ), Candida albicans (C. albicans ), Trichophyton rubrum (T. rubrum) and Trichophyton mentagrophytes (T. mentagrophytes ), which are related to the skin, it was confirmed that the Neem extract treatment showed clear zones in all 9 bacteria at 5 mg/mL concentration and particularly, 8, 7, and 7 mm clear zones were observed from P. acnes, C. xerosis and T. mentagrophytes , respectively. The Comfrey extract treatment also showed clear zones in all 9 bacteria. Especially, it showed 10, 8, 8, 10, and 11 mm clear zones from P. acnes , C. xerosis , P. ovale , T. rubrum and T. mentagrophytes . From these results, it was confirmed that the Comfrey extract had higher antibacterial and antifungal effects than the Neem extract had, and therefore, both the Neem and Comfrey extracts have excellent antibacterial effects against P. acnes , C. xerosis , and T. mentagrophytes among various bacteria. Conclusion: The Neem and the Comfrey extracts had great antioxidative effects as well as antibacterial and antifungal effects against 9 different skin-related bacteria. Therefore, they can be considered to be useful as natural preservatives and cosmetic ingredients.

General anesthesia induces changes in dendritic spine number and synaptic transmission in developing mice. These changes are rather disturbing, as similar changes are seen in animal models of neurodevelopmental disorders. We previously suggested that mTor-dependent upregulation of mitochondrial function may be involved in such changes. To further understand the significance of mitochondrial changes after general anesthesia during neurodevelopment, we exposed young mice to 2.5 % sevoflurane for 2 h followed by injection of rotenone, a mitochondrial complex I inhibitor. In postnatal day 17 (PND17) mice, intraperitoneal injection of rotenone not only blocked sevoflurane-induced increases in mitochondrial function, it also prevented sevoflurane-induced changes in excitatory synaptic transmission. Interestingly, similar changes were not observed in younger, neonatal mice (PND7). We next assessed whether the mitochondrial unfolded protein response (UPRmt) acted as a link between anesthetic exposure and mitochondrial function. Expression of UPRmt proteins, which help maintain protein-folding homeostasis and increase mitochondrial function, was increased 6 h after sevoflurane exposure. Our results show that a single, brief sevoflurane exposure induces age-dependent changes in mitochondrial function that constitute an important mechanism for the increase in excitatory synaptic transmission in late postnatal mice, and also suggest mitochondria and UPRmt as potential targets for preventing anesthesia toxicity.

Purpose: Using extracts from different parts of Zingiber officinale , we aimed to evaluate its antibacterial effect on skin flora and its deodorization effect on ammonia to determine its potential as an antimicrobial ingredient and functional herbal cosmetic ingredient. Methods: The antibacterial effect on skin flora was examined using the paper disc method and the deodorization effect on ammonia was examined by the gas detection method. Results: After evaluating the antibacterial effects of extracts from different parts (leaves, stems, and roots) of Zingiber officinale , antibacterial activity was observed against Staphylococcus aureus (S. aureus ) at a concentration of 5 mg/mL. The leaf, stem, and root extracts had 3 mm, 3 mm, and 2mm clear zones, respectively. For the antibacterial effect against Staphylococcus epidermidis (S. epidermidis ), there were 8 mm, 5 mm, and 4mm clear zones for the leaf, stem, and root extracts, respectively. For Propionibacterium acnes (P. acnes ), a bacterium that causes acne, there were 4 mm, 3 mm, and 1mm clear zones for the leaf, stem, and root extracts, respectively. The results for Pityrosporum ovale (P. ovale ) showed 6 mm, 5 mm, and 4 mm clear zones for the leaf, stem, and root extracts, respectively. For Malassezia furfur (M. furfu ), there were 4 mm, 3 mm, and 2 mm clear zones for the leaf, stem, and root extract, respectively. For Candida albicans (C. albicans ), there were 4 mm, 3 mm, and 2mm clear zones for the leaf, stem, and root extracts, respectively. Finally, after evaluating the antibacterial effect against Trichophyton mentagrophytes (T. mentagrophytes ) at a concentration of 5 mg/mL, we found 6 mm, 4 mm, and 3 mm clear zones for the leaf, stem, and root extracts, respectively. After evaluating the deodorization effects of Zingiber officinale extracts on ammonia, at 120 min, the deodorization effect was 98%, 95%, and 91% for the leaf, stem, and root extracts, respectively. Conclusion: Among the three extracts of Zingiber officinale , the leaf extract had the highest antibacterial effect against the seven types of skin flora as well as the highest deodorization effect, thus confirming its potential as an antibacterial ingredient and functional herbal cosmetic ingredient.

Mitochondrial oxidative phosphorylation (OxPhos) is essential for energy production and survival. However, the tissue-specific and systemic metabolic effects of OxPhos function in adipocytes remain incompletely understood.We used adipocyte-specific Crif1 (also known as Gadd45gip1) knockout (AdKO) mice with decreased adipocyte OxPhos function. AdKO mice fed a normal chow or high-fat diet were evaluated for glucose homeostasis, weight gain and energy expenditure (EE). RNA sequencing of adipose tissues was used to identify the key mitokines affected in AdKO mice, which included fibroblast growth factor 21 (FGF21) and growth differentiation factor 15 (GDF15). For in vitro analysis, doxycycline was used to pharmacologically decrease OxPhos in 3T3L1 adipocytes. To identify the effects of GDF15 and FGF21 on the metabolic phenotype of AdKO mice, we generated AdKO mice with global Gdf15 knockout (AdGKO) or global Fgf21 knockout (AdFKO).Under high-fat diet conditions, AdKO mice were resistant to weight gain and exhibited higher EE and improved glucose tolerance. In vitro pharmacological and in vivo genetic inhibition of OxPhos in adipocytes significantly upregulated mitochondrial unfolded protein response-related genes and secretion of mitokines such as GDF15 and FGF21. We evaluated the metabolic phenotypes of AdGKO and AdFKO mice, revealing that GDF15 and FGF21 differentially regulated energy homeostasis in AdKO mice. Both mitokines had beneficial effects on obesity and insulin resistance in the context of decreased adipocyte OxPhos, but only GDF15 regulated EE in AdKO mice.The present study demonstrated that the adipose tissue adaptive mitochondrial stress response affected systemic energy homeostasis via cell-autonomous and non-cell-autonomous pathways. We identified novel roles for adipose OxPhos and adipo-mitokines in the regulation of systemic glucose homeostasis and EE, which facilitated adaptation of an organism to local mitochondrial stress.

Purpose: This study tests the anti-oxidant effect of mint, cinnamon and their mixed extract and their antibacterial effects on the skin resident flora. This study assessed its stability, and confirmed the adequacy of the component as a material for cosmetics for the scalp by manufacturing a shampoo containing the mixed extract. Methods: Mint and cinnamon were extracted by 70% with ethanol. The mixed extract was prepared by mixing mint and cinnamon by 1:1. As for the antioxidant effects, the followings were measured: DPPH radical scavenging activity, total polyphenol and flavonoid contents, and ABTS radical scavenging activity. The antibacterial effects on the seven types of bacteria in the extract (by paper disc) were analyzed. Finally, a shampoo containing the mixed extract from mint and cinnamon was manufactured and stored for 12 weeks. Its temperature and pH change were measured to assess the stability. Results: Regarding the DPPH radical scavenging activity, the mint extract was 78.63%, cinnamon was 86.25%, and mixed extract was 89.25%. The total polyphenol contents were 80.22±1.03, 101.20±1.64, and 135.27±1.23 μg/mL, and the total flavonoid contents were 26.12±1.01, 56.31±1.13, and 72.65±1.13 μg/mL. As a result of measuring the ABTS radical scavenging activity, the mint extract was 72.20, cinnamon extract 88.12%, and the total mixed extract was 90.21%. The antibacterial effect was confirmed on all bacteria. In the mint extract, the antibacterial effect was high in P. ovale , M. furfur , T. mentagrophytes , and in the cinnamon extract, the effect was high in P. ovale . In the mixed extract, the effect was high in P. ovale and M. furfur . A shampoo containing the mixed extract was manufactured, and its stability and pH change were measured at different temperatures (25℃, 40℃ and 60℃). In the end, the stability was confirmed. Conclusion: The mixed extract of mint and cinnamon had the higher anti-oxidant and antibacterial effects than mint and cinnamon extracts. The measurement of the temperature stability and pH change in the manufactured shampoo showed constant pH and temperature with no significant change, thus confirming the stability of the shampoo. If additional studies can be conducted in the future, the extract can be commercialized as an ingredient for cosmetics for the scalp.

Purpose: The antioxidant effects and antibacterial effects on 7 types of dermatitis of Codonopsis lanceolata (C. lanceolata ) ethanol and water extracts were investigated to study its potential as an ingredient of antibacterial cosmetics for skin and functional cosmetics for hair by testing its deodorization effects on ammonia. Methods: The total polyphenol and flavonoid contents, electron donating ability, ABTS radical scavenging activity of C. lanceolata extracts were investigated for its antioxidant effects and the antibacterial effects of c. lanceolata extract on 7 different germs causing dermatitis were explored with the paper disc method. In addition, a gas detection method was employed to test the deodorizing effects on ammonia. Results: As a result of the antioxidant effects test, the total polyphenol contents of C. lanceolata ethanol and water extracts were 88.86±1.64 and 76.46±1.23 μg/ mL, respectively, and the total flavonoid contents were measured 78.29±1.13 and 60.02±1.35μg/mL, respectively. As for the 2,2-diphenylpicrylhydrazyl (DPPH) scavenging activity was 78.78% and 62.12%. The test on 2,2'-Azinobis [3-ethylbenzothiazoline-6-sulfonic acid]-diammonium salt (ABTS) radical scavenging activity with was 80.12% and 72.24%. For the 7 dermatitis germs, C. lanceolata ethanol extract and water extract showed antibacterial effects at the concentration of 5 mg/mL. For the deodorization effects of C. lanceolata extract on ammonia, ethanol extract revealed 98% of deodorizing effects after 30 min. Conclusion: C. lanceolata ethanol extract had greater antioxidant effects and antibacterial effects on the 7 dermatitis germs than water extract; with higher deodorizing effects, it is considered suitable for making antibacterial cosmetics for skin and functional products for hair.

Cognitive decline is observed in aging and neurodegenerative diseases, including Alzheimer&rsquo, s disease (AD) and dementia. Intracellular energy produced via mitochondrial respiration is used in the regulation of synaptic plasticity and structure, including dendritic spine length and density, as well as for the release of neurotrophic factors involved in learning and memory. To date, a few synthetic agents for improving mitochondrial function have been developed for overcoming cognitive impairment. However, no natural compounds that modulate synaptic plasticity by directly targeting mitochondria have been developed. Here, we demonstrate that a mixture of Schisandra chinensis extract (SCE) and ascorbic acid (AA) improved cognitive function and induced synaptic plasticity-regulating proteins by enhancing mitochondrial respiration. Treatment of embryonic mouse hippocampal mHippoE-14 cells with a 4:1 mixture of SCE and AA increased basal oxygen consumption rate. We found that mice injected with the SCE-AA mixture showed enhanced learning and memory and recognition ability. We further observed that injection of the SCE-AA mixture in mice significantly increased expression of postsynaptic density protein 95 (PSD95), an increase that was correlated with enhanced brain-derived neurotrophic factor (BDNF) expression. These results demonstrate that a mixture of SCE and AA improves mitochondrial function and memory, suggesting that this natural compound mixture could be used to alleviate AD and aging-associated memory decline.

목적: 더덕 껍질 추출물의 생리활성 및 화장품 소재로서의 가능성과 산업적 활용가치를 확인하고자 하였다. 방법: 더덕 껍질 열수 그리고 에탄올 추출물을 이용하여 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical 소거활성 및 총 폴리페놀, 플라보노이드 함량, superoxide dismutase (SOD) 유사활성을 조사하여 항산화 효과를 확인하고, HaCaT 세포에서 H2O2에 대한 세포 보호 효과를 측정 하였다. LPS로 염증반응이 유도된 RAW 264.7 세포를 통해 nitric oxide (NO) 생성 억제 효과를 Griess의 방법으로 조사하였다. 결 과: 항산화 효과 측정 결과, DPPH 소거활성은 500 μg/mL에서 열수 추출물은 74.93%, 에탄올 추출물은 85.25% 소거능이 확인 되 었으며, 열수 및 에탄올 추출물의 총 폴리페놀 함량은 각각 70.22, 75.27 μg/mL 이고, 총 플라보노이드 함량은 16.12, 22.65 μg/ mL으로 확인되었다. SOD 유사활성 측정 결과, 500 μg/mL에서 열수 추출물은 62.20%, 에탄올 추출물은 69.93%의 활성이 확인되 었다. HaCaT 세포에서 H2O2에 대한 세포 보호 효과 측정결과, 100μ g/mL 농도에서 열수 및 에탄올 추출물은 각각 30, 36%의 보호 효과가 확인 되었다. Lipopolysaccharides (LPS)로 염증반응이 유도된 RAW 264.7 세포를 통해 NO 저해 효과 측정 결과, 100 μg/ mL 농도에서 열수 및 에탄올 추출물의 NO 생성량은 각각 14.57 μM, 11.45 μM로 감소하였다. 결론: 더덕 껍질 에탄올 추출물은 열수 추출물보다 항산화 효과 및 세포 보호 효과 그리고 항염증 효과가 더 우수하였으며, 천연 기능성 화장품 소재로서의 가능성이 있다고 판단된다.

목적: 아로니아 잎 추출물의 화장품 기능성 소재로서 활용 가능성을 알아보기 위하여 항산화, 미백, 항염증, collagen 합성량을 측 정하였다. 방법: 아로니아 잎을 에탄올, 열수 추출하여 시료로 사용하였다. 1,1-diphenyl-2-picrylhydrazyl (DPPH) 소거능과 총 폴리페놀 함량, 총 플라보노이드 함량을 측정하였으며, B16F10 mouse melanoma 세포주를 사용하여 미백효과를 측정하였다. 마 우스의 RAW 264.7 대식세포에 lipopolysaccharide (LPS)를 처리하여 nitric oxide (NO) 억제 효과를 확인 하였으며, human dermal fibroblast (HDF)에서 ELISA kit를 이용하여 procollagen 합성량을 측정하였다. 결과: 아로니아 잎 에탄올, 열수 추출물의 총 폴리페 놀 함량은 167.46±1.03, 79.04±1.01 μg/mL으로 확인되었으며, 총 플라보노이드 함량은 105.15±1.22, 52.21±1.13 μg/mL로 나타났다. DPPH radical 소거능은 아로니아 에탄올 추출물의 효과가 높게 확인되었다. B16F10 세포에서 아로니아 잎의 tyrosinase 저해활성 및 melanin 저해율의 경우, 에탄올 추출물이 100 μg/mL 농도에서 21%, 24%의 저해 효과가 확인되었으며, 열수 추출 물은 18%, 18%의 저해 효과가 확인되었다. α-melanocyte-stimulating hormone (α-MSH) 유도에 의한 tyrosinase 저해활성 및 melanin 저해율도 아로니아 에탄올 추출물의 효과가 더 높게 확인되었다. 아로니아 잎의 항염증 효과는 에탄올 추출물의 NO 생성 억제능이 더 우수하였다. 아로나이 잎 에탄올, 열수 추출물의 200 μg/mL 농도에서 각각 58%, 41%의 procollagen 합성이 증가하여 양성대조군 보다는 낮은 procollagen 합성이 확인 되었지만 HDF 세포에서 procollagen 합성을 증가시킴으로써 주름개선 효과를 가 지는 것을 확인 할 수 있었다. 결론: 이상의 결과를 통해 아로니아 잎은 에탄올 추출물이 열수 추출물 보다 항산화, 미백 그리고 항 염증 및 collagen 합성 효과가 더 우수하였으며, 기능성 화장품 소재로서의 활용 가능하리라고 생각된다.

The derangement of tyrosine hydroxylase (TH) activity reduces dopamine synthesis and is implicated in the pathogenesis of Parkinson's disease. However, the extracellular modulator and intracellular regulatory mechanisms of TH have yet to be identified. Recently, high-mobility group box 1 (HMGB1) was reported to be actively secreted from glial cells and is regarded as a mediator of dopaminergic neuronal loss. However, the mechanism for how HMGB1 affects TH expression, particularly through the receptor for advanced glycation endproducts (RAGE), has not yet been investigated. We found that recombinant HMGB1 (rHMGB1) upregulates TH mRNA expression via simultaneous activation of JNK phosphorylation, and this induction of TH expression is blocked by inhibitors of RAGE and JNK. To investigate how TH expression levels change through the HMGB1-RAGE axis as a result of MPP+ toxicity, we co-treated SN4741 dopaminergic cells with MPP+ and rHMGB1. rHMGB1 blocked the reduction of TH mRNA following MPP+ treatment without altering cell survival rates. Our results suggest that HMGB1 upregulates TH expression to maintain dopaminergic neuronal function via activating RAGE, which is dependent on JNK phosphorylation.

Movement defects in obesity are associated with peripheral muscle defects, arthritis, and dysfunction of motor control by the brain. Although movement functionality is negatively correlated with obesity, the brain regions and downstream signaling pathways associated with movement defects in obesity are unclear. A dopaminergic neuronal pathway from the substantia nigra (SN) to the striatum is responsible for regulating grip strength and motor initiation through tyrosine hydroxylase (TH) activity-dependent dopamine release. We found that mice fed a high-fat diet exhibited decreased movement in open-field tests and an increase in missteps in a vertical grid test compared with normally fed mice. This motor abnormality was associated with a significant reduction of TH in the SN and striatum. We further found that phosphorylation of c-Jun N-terminal kinase (JNK), which modulates TH expression in the SN and striatum, was decreased under excess-energy conditions. Our findings suggest that high calorie intake impairs motor function through JNK-dependent dysregulation of TH in the SN and striatum.

Recent studies revealed that the inhibition of mitochondrial oxidative phosphorylation (OXPHOS) is coupled with the mitochondrial unfolded protein response, thereby stimulating the secretion of non-cell autonomous factors, which may control systemic energy metabolism and longevity. However, the nature and roles of non-cell autonomous factors induced in adipose tissue in response to reduced OXPHOS function remain to be clarified in mammals. CR6-interacting factor 1 (CRIF1) is an essential mitoribosomal protein for the intramitochondrial production of mtDNA-encoded OXPHOS subunits. Deficiency of CRIF1 impairs the proper formation of the OXPHOS complex, resulting in reduced function. To determine which secretory factors are induced in response to reduced mitochondrial OXPHOS function, we analyzed gene expression datasets in Crif1-depleted mouse embryonic fibroblasts. Crif1 deficiency preferentially increased the expression of angiopoietin-like 6 (Angptl6) and did not affect other members of the ANGPTL family. Moreover, treatment with mitochondrial OXPHOS inhibitors increased the expression of Angptl6 in cultured adipocytes. To confirm Angptl6 induction in vivo, we generated a murine model of reduced mitochondrial OXPHOS function using adipose tissue-specific Crif1-deficient mice and verified the upregulation of Angptl6 and fibroblast growth factor 21 (Fgf21) in white adipose tissue. Treatment with recombinant ANGPTL6 protein increased oxygen consumption and Pparα expression through the extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in cultured adipocytes. Furthermore, the ANGPTL6-mediated increase in Pparα expression resulted in increased FGF21 expression, thereby promoting β-oxidation. In conclusion, mitochondrial OXPHOS function governs the expression of ANGPTL6, which is an essential factor for FGF21 production in adipose tissue and cultured adipocytes.

목적: 감초 추출물에 대하여 생리활성 및 화장품 소재로서의 가능성과 산업적 활용가치를 확인하고자 하였다. 방법: 국내산 감초를 70% 에탄올 및 열수 추출하여, 항산화, 산화적 스트레스에 대한 HaCaT 세포의 보호 효과, 항염 효능을 분석하고, β-hexosaminidase의 억제량을 측정하여 항알레르기 효능을 확인하였다. 또한 피부 상재균에 대한 항균 활성도 측정하였다. 결과: 항산화력을 비교한 결과, 70% 에탄올 추출물이 열수 추출물보다 우수하였으며, 감초 추출물을 이용한 세포 실험을 진행한 결과 HaCaT, RAW 264.7, RBL-2H3 세포에서 유의한 세포독성은 나타나지 않았다. Hydrogen peroxide (H2O2)에 의해 유발되는 산화적 스트레스에 대한 HaCaT 세포의 보호 효과를 확인한 결과, 70% 에탄올 추출물 100 μg/mL의 농도에서 28%의 보호 효과가 확인되었다. RAW 264.7 세포에 대한 감초 추출물의 항염 효과를 확인한 결과, 저농도에서도 nitric oxide (NO) 생성이 억제되었다. β-Hexosaminidase의 분비를 측정한 결과, 감초 에탄올 추출물과 열수 추출물은 농도 의존적으로 면역세포의 탈과립을 억제하였다. Staphylococcus aureus (S. aureus ), Staphylococcus epidermidis (S. epidermidis ), Propionibacterium acnes (P.acnes ) 균에서도 감초 추출물은 농도 의존적으로 항균 활성이 확인되었다. 결론: 본 연구는 감초 추출물의 기능성 화장품 소재로 사용 시 민감성 화장품 소재로서 사용하는 것을 제안하며 향후 임상적 활용 가능성을 판단하기 위하여 동물, 인체시험을 통한 감초 추출물의 효능 검증 연구가 체계적으로 진행되어야 할 것이다.

Cerebral endothelial cells (ECs) require junctional proteins to maintain blood–brain barrier (BBB) integrity, restricting toxic substances and controlling peripheral immune cells with a higher concentration of mitochondria than ECs of peripheral capillaries. The mechanism underlying BBB disruption by defective mitochondrial oxidative phosphorylation (OxPhos) is unclear in a mitochondria-related gene-targeted animal model. To assess the role of EC mitochondrial OxPhos function in the maintenance of the BBB, we developed an EC-specific CR6-interactin factor1 ( Crif1) deletion mouse. We clearly observed defects in motor behavior, uncompacted myelin and leukocyte infiltration caused by BBB maturation and disruption in this mice. Furthermore, we investigated the alteration in the actin cytoskeleton, which interacts with junctional proteins to support BBB integrity. Loss of Crif1 led to reorganization of the actin cytoskeleton and a decrease in tight junction-associated protein expression through an ATP production defect in vitro and in vivo. Based on these results, we suggest that mitochondrial OxPhos is important for the maturation and maintenance of BBB integrity by supplying ATP to cerebral ECs.

Primary refractory acute myeloid leukemia (AML) and early recurrence of leukemic cells are among the most difficult hurdles to overcome in the treatment of AML. Moreover, uncertainties surrounding the molecular mechanism underlying refractory AML pose a challenge when it comes to developing novel therapeutic drugs. However, accumulating evidence suggests a contribution of phosphatase and tensin homolog (PTEN)/protein kinase B (AKT) signaling to the development of refractory AML. To assess PTEN/AKT signaling in AML, two types of AML cell lines were evaluated, namely control HL60 cells and KG1α cells, a refractory AML cell line that is resistant to idarubicin and cytarabine (AraC) treatment. Changes in the expression level of glycolysis‑ and mitochondrial oxidative phosphorylation‑related genes and proteins were evaluated by reverse transcription‑quantitative polymerase chain reaction and western blot analyses, respectively. The mitochondrial oxygen consumption and extracellular acidification rates were measured using an XF24 analyzer. CCK8 assay and Annexin V/PI staining were used to analyze cell viability and cellular apoptosis, respectively. The PTEN protein was found to be depleted, whereas AKT phosphorylation levels were elevated in KG1α cells compared with HL60 cells. These changes were associated with increased expression of glucose transporter 1 and hexokinase 2, and increased lactate production. AKT inhibition decreased the proliferation of KG1α cells and decreased extracellular acidification without affecting HL60 cells. Notably, AKT inhibition increased the susceptibility of KG1α cells to chemotherapy with idarubicin and AraC. Taken together, the findings of the present study indicate that activation of AKT by PTEN deficiency sustains the refractory AML status through enhancement of glycolysis and mitochondrial respiration, effects that may be rescued by inhibiting AKT activity.

Mild cognitive impairment (MCI) is considered as a transitional stage between aging and Alzheimer&rsquo, s disease. In the present study, we examined the protective effect of Schisandra chinensis (SC) and Ribes fasciculatum (RF) on neuronal cell death in vitro and scopolamine-induced cognitive impairment in Sprague Dawley&reg, rats in vivo. A mixture of SC and RF extracts (SC+RF) significantly protected against hydrogen peroxide-induced PC12 neuronal cell death. The neuroprotective effect of SC+RF on scopolamine-induced memory impairment in rats was evaluated using the passive avoidance test and the Morris water maze test. In the passive avoidance test, SC+RF-treated rats showed an increased latency to escape, compared to the scopolamine-treated rats. Moreover, SC+RF treatment significantly reduced escape latency in water maze test, compared to treatment with scopolamine alone. To verify the long-term memory, we performed probe test of water maze test. As a result, rat treated with SC+RF spent more time in the target quadrant. Consistent with enhancement of memory function, the brain derived neurotrophic factor (BDNF) and its downstream molecules (pERK, pATK, and pCREB) are increased in SC+RF treatment in hippocampal area compared with scopolamine treated group. These results suggest that a mixture of SC and RF extracts may be a good therapeutic candidate for preventing mild cognitive impairment.

Current therapeutics for Parkinson&rsquo, s disease (PD) are only effective in providing relief of symptoms such as rigidity, tremors and bradykinesia, and do not exert disease-modifying effects by directly modulating mitochondrial function. Here, we investigated auraptene (AUR) as a potent therapeutic reagent that specifically protects neurotoxin-induced reduction of mitochondrial respiration and inhibits reactive oxygen species (ROS) generation. Further, we explored the mechanism and potency of AUR in protecting dopaminergic neurons. Treatment with AUR significantly increased the viability of substantia nigra (SN)-derived SN4741 embryonic dopaminergic neuronal cells and reduced rotenone-induced mitochondrial ROS production. By inducing antioxidant enzymes AUR treatment also increased oxygen consumption rate. These results indicate that AUR exerts a protective effect against rotenone-induced mitochondrial oxidative damage. We further assessed AUR effects in vivo, investigating tyrosine hydroxylase (TH) expression in the striatum and substantia nigra of MPTP-induced PD model mice and behavioral changes after injection of AUR. AUR treatment improved movement, consistent with the observed increase in the number of dopaminergic neurons in the substantia nigra. These results demonstrate that AUR targets dual pathogenic mechanisms, enhancing mitochondrial respiration and attenuating ROS production, suggesting that the preventative potential of this natural compound could lead to improvement in PD-related neurobiological changes.

목적: 본 연구에서는 갈대, 함초, 매실 에탄올 추출물의 항산화 효과, 항알레르기 효과를 확인하여 기능성 소재 및 화장품 소재로의개발 가능성을 확인하고자 하였다. 방법: 갈대, 함초, 매실을 70% 주정 에탄올로 추출하였으며, 각각 추출물의 산화적 스트레스 요인에 대한 인간 피부세포유래인HaCaT 세포주의 보호효과를 측정하였다. 항알레르기 효과를 확인하고자 알레르기 반응을 나타내는 주요 세포인 비만세포 RBL- 2H3 세포주를 이용하여 알레르기 반응 시 세포의 탈과립 지표로 알려진 β-hexosaminidase 분비 억제를 관찰하였다. 결과: 활성산소에 의한 산화적 스트레스로부터 세포를 보호할 수 있는 항산화력을 평가한 결과 세 가지 추출물 중 갈대 에올탄 추출물의경우 200 μg/mL의 농도에서 가장 높은 37%의 보호효과를 나타내었다. β-Hexosaminidase 분비에 대한 갈대, 함초, 매실 에탄올추출물의 억제 효능을 확인하였다. 추출물을 처리하지 않은 대조군과 비교하였을 때, 갈대, 함초, 매실 에탄올 추출물은 각각 200 μg/mL의 농도에서 각각 53.6%, 49.8%, 30.2%의 효과를 나타내었고, 세 가지의 추출물 중 갈대의 효과가 가장 우수했다. 결론: 갈대, 함초, 매실 에탄올 추출물은 세포에 대한 독성을 갖지 않으면서 산화적 스트레스에 대한 항산화와 가려움증을억 제하는효과가 있음을 확인하였기 때문에 기능성 소재 및 화장품 소재로 개발 가능성이 있을 것으로 판단된다.

Long-term imatinib treatment induces drug-resistant chronic myeloid leukemia (CML) cells harboring T315I gate keeper mutation of breakpoint cluster region (BCR)-ABL oncogenic kinase. However, although cell proliferation is coupled with cellular energy status in CML carcinogenesis, the metabolic characteristics of T315I-mutant CML cells have never been investigated. Here, we analyzed cell proliferation activities and metabolic phenotypes, including cell proliferation, oxygen consumption, lactate production, and redox state in the KBM5 (imatinib-sensitive) and KBM5-T315I (imatinib-resistant) CML cell lines. Interestingly, KBM5-T315I cells showed decreased cell proliferation, lactate production, fatty acid synthesis, ROS production, and down regulation of mRNA expression related to ROS scavengers, such as SOD2, catalase, GCLm, and GPx1. Taken together, our data demonstrate that the lower growth ability of KBM5-T315I CML cells might be related to the decreased expression of glycolysis-related genes and ROS levels, and this will be used to identify therapeutic targets for imatinib resistance in CML.

Paraquat (PQ), an herbicide considered an environmental contributor to the development of Parkinson’s disease (PD), induces dopaminergic neuronal loss through reactive oxygen species (ROS) production and oxidative stress by mitochondrial complex I. Most patients with PQ-induced PD are affected by chronic exposure and require a preventive strategy for modulation of disease progression. To identify drugs that are effective in preventing PD, we screened more than 1000 drugs that are currently used in clinics and in studies employing PQ-treated cells. Of these, chloramphenicol (CP) showed the most powerful inhibitory effect. Pretreatment with CP increased the viability of PQ-treated SN4741 dopaminergic neuronal cells and rat primary cultured dopaminergic neurons compared with control cells treated with PQ only. CP pretreatment also reduced PQ-induced ROS production, implying that mitochondrial complex I is a target of CP. This effect of CP reflected downregulation of the mitochondrial complex I subunit ND1 and diminished PQ recycling, a major mechanism of ROS production, and resulted in the prevention of cell loss. Notably, these effects of CP were not observed in rotenone-pretreated SN4741 cells and Rho-negative cells, in which mitochondrial function is defective. Consistent with these results, CP pretreatment of MPTP-treated PD model mice also ameliorated dopaminergic neuronal cell loss. Our findings indicate that the inhibition of mitochondrial complex I with CP protects dopaminergic neurons and may provide a strategy for preventing neurotoxin-induced PD.

This work was supported by grants from the NIH (R01DK083567 to YBK), the American Diabetes Association (1-09-RA-87 to YBK), the American Heart Association (12GRANT12040170 to YBK), the East Carolina University Start-up fund (to HH), the National Research Foundation (NRF-2014M3A9D8034464 to M Shong, NRF-2016R1A2B3010373 to KSP, NRF-2015R1C1A1A02037164 to SHL), the National Research Foundation of Korea (2013M3C7A1056024 to MSK), and the Korean Diabetes Association (to JAS, 2017). In addition, structural funding for the Center for Neuroscience and Cell Biology, University of Coimbra, NMR facility is supported by FEDER-PT2020 (UID/BIA/04004/2013 and CENTRO-07-CT62-FEDER-002012) and by the Portuguese Foundation for Science and Technology (FCT) through grants PTDC/CVT-NUT/2851/2014, PTDC/BIM-MET/4265/2014, and RECI/QEQ-QFI/0168/2012. ISL is a recipient of an FCT fellowship from Portugal (SFRH/BD/71021/2010), and MCK is a recipient of a postdoctoral fellowship award from the American Diabetes Association (1-17-PDF-146). GDB is supported by the European Union's Horizon 2020 Research and Innovation programme under Marie Sklodowska-Curie Grant Agreement 722619. We thank Farhad Danesh for CA-ROCK1-knockin mice; Huseyin Ozkan, Ivan Viegas, Cristina Barosa, Hyun Cheol Rho, Xuemei Ma, Yao Yang, and Alexander Banks for technical help; and Barbara Kahn, Tony Hollenberg, Sonia Najjar, and Terry Flier for helpful discussion. Obesity is a major risk factor for developing nonalcoholic fatty liver disease (NAFLD). NAFLD is the most common form of chronic liver disease and is closely associated with insulin resistance, ultimately leading to cirrhosis and hepatocellular carcinoma. However, knowledge of the intracellular regulators of obesity-linked fatty liver disease remains incomplete. Here we showed that hepatic Rho-kinase 1 (ROCK1) drives obesity-induced steatosis in mice through stimulation of de novo lipogenesis. Mice lacking ROCK1 in the liver were resistant to diet-induced obesity owing to increased energy expenditure and thermogenic gene expression. Constitutive expression of hepatic ROCK1 was sufficient to promote adiposity, insulin resistance, and hepatic lipid accumulation in mice fed a high-fat diet. Correspondingly, liver-specific ROCK1 deletion prevented the development of severe hepatic steatosis and reduced hyperglycemia in obese diabetic (ob/ob) mice. Of pathophysiological significance, hepatic ROCK1 was markedly upregulated in humans with fatty liver disease and correlated with risk factors clustering around NAFLD and insulin resistance. Mechanistically, we found that hepatic ROCK1 suppresses AMPK activity and a ROCK1/AMPK pathway is necessary to mediate cannabinoid-induced lipogenesis in the liver. Furthermore, treatment with metformin, the most widely used antidiabetes drug, reduced hepatic lipid accumulation by inactivating ROCK1, resulting in activation of AMPK downstream signaling. Taken together, our findings establish a ROCK1/AMPK signaling axis that regulates de novo lipogenesis, providing a unique target for treating obesity-related metabolic disorders such as NAFLD. publishersversion published

The identification of large numbers of genetic mutations in immature myeloid cells has made it difficult to identify specific targets for acute myeloid leukemia (AML) therapy. Although current pharmacological targets for controlling cancer are focused on identifying genetic mutations, it is hard to develop the specific drugs to achieve complete remission due to complex and variable genetic mutations. To overcome the failure of the genetic mutation theory, the present study targeted mitochondrial metabolism as a strategy for inducing anti‑leukemic activity, based on evidence that AML cells have an abnormally high amount of mitochondria and that somatic mutations can alter metabolic flux in cancer. It was found that L‑deprenyl, which is clinically available for the treatment of Parkinson's disease, exerts anti‑mitochondria activity in KG‑1α cells, as assessed by detection of oxygen consumption rate (OCR) and extracellular acidification (ECAR) using XF analyzer, respectively. Using a luciferase assay for detecting adenosine triphosphate (ATP) content, it was found that suppression of mitochondrial activity led to ATP depletion and was associated with potent cytotoxic activity. L‑deprenyl is known to target monoamine oxidase‑B (MAO‑B) on the outer membrane of mitochondria, therefore, the activity of MAO‑A and ‑B was measured based on the fluorometric detection of H2O2 produced by the enzyme reaction. Notably, MAO‑A and -B activity was low in AML cells and the present findings suggested that the anticancer effect of L‑deprenyl was independent of MAO‑B. Change of mitochondrial respiration‑ and glycolysis‑related gene expression levels were measured by reverse transcription‑quantitative polymerase chain reaction. Consistent with the aforementioned results, treatment with L‑deprenyl reduced the mRNA level of mitochondrial respiration‑ and glycolysis‑related genes. Collectively, the present results identify L‑deprenyl as a novel candidate for the treatment of AML through inhibition of mitochondrial respiration.

High-mobility group box 1 (HMGB1) is actively secreted from inflammatory cells and acts via a non-cell-autonomous mechanism to play an important role in mediating cell proliferation and migration. The HMGB1-RAGE (receptor for advanced glycation end products) axis upregulates tyrosine hydroxylase (TH) expression in response to extracellular insults in dopaminergic neurons in vitro, but little is known about HMGB1 in modulation of dopaminergic neurons in vivo. Here, using immunohistochemistry, we show that HMGB1 and RAGE expression are higher in the nigral area of MPTP (methyl-4-phenyl-1,2,3,6-tetrahydropyridine)-treated mice, a toxin-induced Parkinsonian mouse model, compared with saline-treated controls. HMGB1 was predominantly localized to astrocytes and may affect neighboring dopaminergic neurons in the MPTP mouse model, owing to co-localization of RAGE in these TH-positive cells. In addition, MPTP induced a decrease in TH expression, an effect that was potentiated by inhibition of c-Jun N-terminal kinase (JNK) or RAGE. Moreover, stereotaxic injection of recombinant HMGB1 attenuated the MPTP-induced reduction of TH in a Parkinsonian mouse model. Collectively, our results suggest that an increase of HMGB1, released from astrocytes, upregulates TH expression in an acute MPTP-induced Parkinsonian mouse model, thereby maintaining dopaminergic neuronal functions.

Renal cell carcinoma (RCC) progression resulting from the uncontrolled migration and enhanced angiogenesis is an obstacle to effective therapeutic intervention. Tumor metabolism has distinctive feature called Warburg effect, which enhances the aerobic glycolysis rapidly supplying the energy for migration of tumor. To manipulate this metabolic change characteristic of aggressive tumors, we utilized the citrus extract, auraptene, known as a mitochondrial inhibitor, testing its anticancer effects against the RCC4 cell line. We found that auraptene impaired RCC4 cell motility through reduction of mitochondrial respiration and glycolytic pathway-related genes. It also strongly disrupted VEGF-induced angiogenesis in vitro and in vivo. Hypoxia-inducible factor 1a (HIF-1a), a key regulator of cancer metabolism, migration and angiogenesis that is stably expressed in RCCs by virtue of a genetic mutation in the von Hippel-Lindau (VHL) tumor-suppressor protein, was impeded by auraptene, which blocked HIF-1a translation initiation without causing cytotoxicity. We suggest that blockade HIF-1a and reforming energy metabolism with auraptene is an effective approach for suspension RCC progression.

Hypothalamic regulation of appetite governs whole-body energy balance. Satiety is regulated by endocrine factors, including leptin, and impaired leptin induction causes obesity. Angiopoietin-like growth factor (AGF) promotes energy expenditure in the periphery, and systemic reconstitution of AGF antagonizes obesity. However, whether hypothalamic AGF plays a role in controlling food intake remains unknown. Here, we demonstrated that AGF is expressed in proopiomelanocortin (POMC)-positive neurons located in the arcuate nucleus (ARC) of the hypothalamus. AGF expression was stimulated by leptin-induced STAT3 phosphorylation. Notably, intracerebroventricular injection of AGF significantly reduced food intake by stimulating phosphorylation of CREB in the POMC and increasing α-melanocyte-stimulating hormone (α-MSH) content in the hypothalamus. We also found that hypothalamic injection of AGF significantly suppressed food intake and decreased body weight in high-fat-diet-induced obese mice, which exhibit leptin insensitivity. Collectively, our findings demonstrate that hypothalamic AGF provokes the anorectic melanocortin pathway and mediates leptin signaling to prevent obesity. Funding: This work was supported by the National Research Foundation of South of Korea (NRF) grant funded by the Ministry of Science, ICT & Future Planning (MSIP) (2017R1E1A1A01075126, 2014R1A6A1029617, 2016R1A2B4010398, 2016R1D1A1B03932766, 2017R1A5A201538, 2017R1A6A3A11029367) and research fund of Chungnam National University Declaration of Interest: The authors have declared that no conflict of interests existed. Ethical Approval: Animal experiments were approved by the Institutional Animal Care and Use Committee of Chungnam National University (ethical approval number, CNU-00912). The hypothalamus was obtained from postmortem brains of otherwise healthy individuals at the University of Miyazaki, Japan, with the approval of institutional review board (approval number, C-0037; May 11, 2018). Written informed consent was acquired from each outpatient, and the study was conducted according to provisions of the Declaration of Helsinki.

Background & Aims Nonalcoholic steatohepatitis (NASH) is associated with cirrhosis and hepatocellular carcinoma. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) play key roles in the development of the disease. However, the therapeutic target of NASH has not been fully defined and new treatments are needed. We investigated the protective effects of the antioxidant indole-derived NecroX-7 in a NASH mouse model using leptin-deficient ob/ob and methionine- and choline-deficient (MCD) diet-fed ob/ob mice. Methods Six-week-old male mice were divided into three groups: ob/+ mice, ob/ob mice treated with vehicle and ob/ob mice treated daily with NecroX-7 (20 mg/kg) for 4 weeks. To study the effects of NecroX-7 in a fibrosis model, NASH was induced by feeding ob/ob mice an MCD diet. The effects of NecroX-7 on NASH progression were evaluated using biochemical, histological and molecular markers. Results NecroX-7-treated ob/ob mice had a marked decrease in serum aspartate aminotransferase and alanine transaminase compared with vehicle-treated controls. Interestingly, hepatic steatosis and lipid peroxidation were significantly improved by NecroX-7 treatment. NecroX-7 inhibited tert-butylhydroperoxide- and H2O2-induced mitochondrial ROS/RNS in primary hepatocytes and attenuated mitochondrial dysfunction in vitro and in vivo. Furthermore, NecroX-7-treated mice exhibited fewer infiltrating macrophages and reduced hepatic tumour necrosis factor-alpha expression. Hepatic fibrosis in MCD-fed ob/ob mice was significantly decreased by NecroX-7 treatment. Conclusions NecroX-7 treatment improved hepatic steatosis and fibrosis in murine NASH models. These effects occurred through the suppression of whole-cell ROS/RNS and inflammatory responses and suggest that NecroX-7 has a potential therapeutic benefit in steatohepatitis.

T-helper type 2 (Th2) cytokines, including interleukin (IL)-13 and IL-4, produced in adipose tissue, are critical regulators of intra-adipose and systemic lipid and glucose metabolism. Furthermore, IL-13 is a potential therapy for insulin resistance in obese mouse models. Here, we examined mediators produced by adipocytes that are responsible for regulating systemic glucose homeostasis in response to Th2 cytokines. We used RNA sequencing data analysis of cultured adipocytes to screen factors secreted in response to recombinant IL-13. Recombinant IL-13 induced expression of growth differentiation factor 15 (GDF15) via the Janus kinase-activated STAT6 pathway. In vivo administration of α-galactosylceramide or IL-33 increased IL-4 and IL-13 production, thereby increasing GDF15 levels in adipose tissue and in plasma of mice; however, these responses were abrogated in STAT6 knockout mice. Moreover, administration of recombinant IL-13 to wild-type mice fed a high-fat diet (HFD) improved glucose intolerance; this was not the case for GDF15 knockout mice fed the HFD. Taken together, these data suggest that GDF15 is required for IL-13–induced improvement of glucose intolerance in mice fed an HFD. Thus, beneficial effects of Th2 cytokines on systemic glucose metabolism and insulin sensitivity are mediated by GDF15. These findings open up a potential pharmacological route for reversing insulin resistance associated with obesity.

Mitochondrial dysfunction has emerged as a major contributing factor to endothelial dysfunction and vascular disease, but the key mechanisms underlying mitochondrial dysfunction-induced endothelial dysfunction remain to be elucidated. In this study, we aim at determining whether mitochondrial dysfunction in endothelial cells plays a key role in vascular disease, by examining the phenotype of endothelial-specific CR6-interacting factor 1 (CRIF1) knockout mice. We also used siRNA-mediated downregulation of CRIF1 gene in the endothelial cells to study about the in vitro pathophysiological underlying mechanisms.Downregulation of CRIF1 in endothelial cells caused disturbances of mitochondrial oxidative phosphorylation complexes and membrane potential, leading to enhanced mitochondrial reactive oxygen species production. Gene silencing of CRIF1 results in decreased SIRT1 expression along with increased endothelial nitric oxide synthase (eNOS) acetylation, leading to reduced nitric oxide production both in vitro and in vivo. Endothelium-dependent vasorelaxation of aortic rings from CRIF1 knockout (KO) mice was considerably less than in wild-type mice, and it was partially recovered by Sirt1 overexpression in CRIF1 KO mice.Our results show for the first time a relationship between mitochondrial dysfunction and impaired vascular function induced in CRIF1 deficiency conditions and also the possible underlying pathway involved.These findings indicate that CRIF1 plays an important role in maintaining mitochondrial and endothelial function through its effects on the SIRT1-eNOS pathway. Antioxid. Redox Signal. 27, 234-249.

Department of Biochemistry, Chungnam national University School of Medicine, Daejeon 301-721, KoreaReceived August 13, 2012, Accepted October 4, 2012Metabolic analysis of CR6 interacting factor 1 (Crif1) deficient mouse embryonic fibroblasts with impairedoxidative phosphorylation has been carried out using LC-MS/MS and GC-MS methods. Metabolic profiles ofthe Crif1 deficient cells were comprehensively obtained for the first time. Loss of oxidative phosphorylationfunctions in mitochondria resulted in cancer-like metabolic reprogramming with consumption of majority ofglucose carbon from up-regulated glycolysis to produce lactate, suppressed utilization of glucose carbon in theTCA cycle, increased amounts of amino acids. The changes in metabolic profile of the Crif1 deficient cells aremost probably a consequence of metabolic reprogramming to meet the needs of energy balance and anabolicprecursors in compensation for the loss of major oxidative phosphorylation functions.Key Words : CRIF1, Metabolic analysis, Mass spectrometryIntroductionCR6 interacting factor 1 (CRIF1) has been originallydiscovered as nuclear protein playing a role of cell cycleregulator, growth regulator and transcriptional cofactor.

Background Anesthesia during the synaptogenic period induces dendritic spine formation, which may affect neurodevelopment. The authors, therefore, evaluated whether changes in synaptic transmission after dendritic spine formation induced by sevoflurane were associated with long-term behavioral changes. The effects of sevoflurane on mitochondrial function were also assessed to further understand the mechanism behind spinogenesis. Methods Postnatal day 16 to 17 mice were exposed to sevoflurane (2.5% for 2 h), and synaptic transmission was measured in the medial prefrontal cortex 6 h or 5 days later. The expression of postsynaptic proteins and mitochondrial function were measured after anesthesia. Long-term behavioral changes were assessed in adult mice. Results Sevoflurane increased the expression of excitatory postsynaptic proteins in male and female mice (n = 3 to 5 per group). Sevoflurane exposure in male mice transiently increased miniature excitatory postsynaptic current frequency (control: 8.53 ± 2.87; sevoflurane: 11.09 ± 2.58) but decreased miniature inhibitory postsynaptic current frequency (control: 10.18 ± 4.66; sevoflurane: 6.88 ± 2.15). Unexpectedly, sevoflurane increased miniature inhibitory postsynaptic current frequency (control: 1.81 ± 1.11; sevoflurane: 3.56 ± 1.74) in female mice (neurons, n = 10 to 21 per group). Sevoflurane also increased mitochondrial respiration in male mice (n = 5 to 8 per group). However, such changes from anesthesia during the critical period did not induce long-term behavioral consequences. Values are presented as mean ± SD. Conclusions Sevoflurane exposure during the critical period induces mitochondrial hyperactivity and transient imbalance of excitatory/inhibitory synaptic transmission, without long-lasting behavioral consequences. Further studies are needed to confirm sexual differences and to define the role of mitochondrial activity during anesthesia-induced spine formation.

SummaryAlthough substantial progress has been made in understanding the mechanisms underlying the expression of mtDNA-encoded polypeptides, the regulatory factors involved in mitoribosome-mediated synthesis and simultaneous insertion of mitochondrial oxidative phosphorylation (OXPHOS) polypeptides into the inner membrane of mitochondria are still unclear. In the present study, disruption of the mouse Crif1 gene, which encodes a mitochondrial protein, resulted in a profound deficiency in OXPHOS caused by the disappearance of OXPHOS subunits and complexes in vivo. CRIF1 was associated with large mitoribosomal subunits that were located close to the polypeptide exit tunnel, and the elimination of CRIF1 led to both aberrant synthesis and defective insertion of mtDNA-encoded nascent OXPHOS polypeptides into the inner membrane. CRIF1 interacted with nascent OXPHOS polypeptides and molecular chaperones, e.g., Tid1. Taken together, these results suggest that CRIF1 plays a critical role in the integration of OXPHOS polypeptides into the mitochondrial membrane in mammals.

There are two causes of Parkinson's disease (PD): environmental insults and genetic mutations of PD-associated genes. Environmental insults and genetic mutations lead to mitochondrial dysfunction, and a combination of mitochondrial dysfunction and increased oxidative stress in dopaminergic neurons is thought to contribute to the pathogenesis of PD. Among the PD-associated genes, DJ-1 acts as a redox sensor for oxidative stress and has been also proposed to maintain mitochondrial complex I activity. To understand molecular functions of DJ-1 in the cell, we have generated DJ-1 null cells from the DJ-1(-/-) mouse embryos. Using these null cells, we investigated the susceptibility to an environmental toxin, paraquat, which is known to inhibit mitochondrial complex I. Interestingly, we found that DJ-1 null cells showed a resistance to paraquat-induced apoptosis, including reduced poly (ADP-ribose) polymerase and procaspase-3. Also DJ-1 null cells generated less superoxide than SN4741 cells by paraquat treatment. Consistent with the reduced paraquat sensitivity, DJ-1 null cells showed reduced complex I activity, which was partially rescued by ectopic DJ-I expression. In summary, our results suggest that DJ-1 is critical to maintain mitochondrial complex I and complex I could be a key target in interaction of paraquat toxicity and DJ-1 for giving rise to PD.

Context: The oncogenic BRAFV600E mutation results in an active structural conformation characterized by greatly elevated ERK activity. However, additional cellular effects caused by subcellular action of BRAFV600E remain to be identified. Objective: To explore these effects, differences in the subcellular localization of wild-type and mutant BRAF in thyroid cancer were investigated. Results: A significant proportion of endogenous and exogenous BRAFV600E, but not wild-type BRAF, was detected in the mitochondrial fraction, similar to other BRAF mutants including BRAFV600D, BRAFV600K, BRAFV600R, and BRAFG469A, which showed elevated kinase activity and mitochondrial localization. Induced expression of BRAFV600E suppressed the apoptotic responses against staurosporine and TNFα/cycloheximide. Interestingly, the mitochondrial localization and antiapoptotic activities of BRAFV600E were unaffected by sorafenib and U0126 suppression of MAPK kinase (MEK) and ERK activities. Similarly, although the RAF inhibitor sorafenib effectively inhibited MEK/ERK activation, it did not block the mitochondrial localization of BRAFV600E. In addition, inducible expression of BRAFV600E increased the glucose uptake rate and decreased O2 consumption, suggesting that BRAFV600E reduces mitochondrial oxidative phosphorylation, a signature feature of cancer cells. Again, these metabolic alterations resulted by BRAFV600E expression were not affected by the treatment of thyroid cells by sorafenib. Therefore, RAF and MEK inhibitors are unable to block the antiapoptotic activity of BRAFV600E or correct the high glucose uptake rate and glycolytic activity and suppressed mitochondrial oxidative phosphorylation induced by BRAFV600E. Conclusions: The mitochondrial localization observed in oncogenic BRAF mutants might be related to their altered responses to apoptotic stimuli and characteristic metabolic phenotypes found in thyroid cancer. The inability of MEK and RAF inhibitors, U0126 and sorafenib, respectively, to block the mitochondrial localization of BRAFV600E has additional therapeutic implications for BRAFV600E-positive thyroid cancers.

The roles of adipokines, proinflammatory cytokines, and adipose tissue macrophages in obesity-associated insulin resistance have been explored in both animal and human studies. However, our current understanding of obesity-associated insulin resistance relies on studies of artificial metabolic extremes. The purpose of this study was to explore the roles of adipokines, proinflammatory cytokines, and adipose tissue macrophages in human patients with modest obesity and early metabolic dysfunction. We obtained omental adipose tissue and fasting blood samples from 51 females undergoing gynecologic surgery. We investigated serum concentrations of proinflammatory cytokines and adipokines as well as the mRNA expression of proinflammatory and macrophage phenotype markers in visceral adipose tissue using ELISA and quantitative RT-PCR. We measured adipose tissue inflammation and macrophage infiltration using immunohistochemical analysis. Serum levels of adiponectin and leptin were significantly correlated with HOMA-IR and body mass index. The levels of expression of MCP-1 and TNF-α in visceral adipose tissue were also higher in the obese group (body mass index ≥ 25). The expression of mRNA MCP-1 in visceral adipose tissue was positively correlated with body mass index (r = 0.428, p = 0.037) but not with HOMA-IR, whereas TNF-α in visceral adipose tissue was correlated with HOMA-IR (r = 0.462, p = 0.035) but not with body mass index. There was no obvious change in macrophage phenotype or macrophage infiltration in patients with modest obesity or early metabolic dysfunction. Expression of mRNA CD163/CD68 was significantly related to mitochondrial-associated genes and serum inflammatory cytokine levels of resistin and leptin. These results suggest that changes in the production of inflammatory biomolecules precede increased immune cell infiltration and induction of a macrophage phenotype switch in visceral adipose tissue. Furthermore, serum resistin and leptin have specific roles in the regulation of adipose tissue macrophages in patients with modest obesity or early metabolic dysfunction.

Among the primary brain tumors, glioblastoma multiforme (GBM) has a radical proliferation ability that complicates the therapeutic modulation of cancer progression. The majority of GBM patients have a low survival rate (

Adult thyroid dysfunction is a common endocrine disorder associated with an increased risk of cardiovascular disease and mortality. A recent epidemiologic study revealed a link between obesity and increased prevalence of hypothyroidism. It is conceivable that excessive adiposity in obesity might lead to expansion of the interfollicular adipose (IFA) depot or steatosis in thyroid follicular cells (thyroid steatosis, TS). In this study, we investigated the morphological and functional changes in thyroid glands of obese humans and animal models, diet-induced obese (DIO), ob/ob, and db/db mice. Expanded IFA depot and TS were observed in obese patients. Furthermore, DIO mice showed increased expression of lipogenesis-regulation genes, such as sterol regulatory element binding protein 1 (SREBP-1), peroxisome proliferator-activated receptor γ (PPARγ), acetyl coenzyme A carboxylase (ACC), and fatty acid synthetase (FASN) in the thyroid gland. Steatosis and ultrastructural changes, including distension of the endoplasmic reticulum (ER) and mitochondrial distortion in thyroid follicular cells, were uniformly observed in DIO mice and genetically obese mouse models, ob/ob and db/db mice. Obese mice displayed a variable degree of primary thyroid hypofunction, which was not corrected by PPARγ agonist administration. We propose that systemically increased adiposity is associated with characteristic IFA depots and TS and may cause or influence the development of primary thyroid failure.

PURPOSE NAD(P)H:Quinone Oxidoreductase 1 (NQO1) C609T missense variant (NQO1*2) and 29 basepair (bp)-insertion/deletion (I29/D) polymorphism of the NRH:Quinone Oxidoreductase 2 (NQO2) gene promoter have been proposed as predictive and prognostic factors for cancer development and progression. The purpose of this study is to investigate the relationship between NQO1/NQO2 genotype and clinico-pathological features of papillary thyroid microcarcinoma (PTMC). MATERIALS AND METHODS Genomic DNA was isolated from 243 patients; and clinical data were retrospectively analyzed. NQO1*2 and tri-allelic polymorphism of NQO2 were investigated by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis. RESULTS PTMC with NQO1*2 frequently exhibited extra-thyroidal extension as compared to PTMC with wild-type NQO1 (p=0.039). There was a significant relationship between I29/I29 homozygosity of NQO2 and lymph node metastasis (p=0.042). Multivariate analysis showed that the I29/I29 genotype was associated with an increased risk of lymph node metastasis (OR, 2.24; 95% CI, 1.10-4.56; p=0.026). CONCLUSION NQO1*2 and I29 allele of the NQO2 are associated with aggressive clinical phenotypes of PTMC, and the I29 allele represents a putative prognostic marker for PTMC.

Impaired mitochondrial oxidative phosphorylation (OXPHOS) has been proposed as an etiological mechanism underlying insulin resistance. However, the initiating organ of OXPHOS dysfunction during the development of systemic insulin resistance has yet to be identified. To determine whether adipose OXPHOS deficiency plays an etiological role in systemic insulin resistance, the metabolic phenotype of mice with OXPHOS–deficient adipose tissue was examined. Crif1 is a protein required for the intramitochondrial production of mtDNA–encoded OXPHOS subunits; therefore, Crif1 haploinsufficient deficiency in mice results in a mild, but specific, failure of OXPHOS capacity in vivo. Although adipose-specific Crif1-haploinsufficient mice showed normal growth and development, they became insulin-resistant. Crif1-silenced adipocytes showed higher expression of chemokines, the expression of which is dependent upon stress kinases and antioxidant. Accordingly, examination of adipose tissue from Crif1-haploinsufficient mice revealed increased secretion of MCP1 and TNFα, as well as marked infiltration by macrophages. These findings indicate that the OXPHOS status of adipose tissue determines its metabolic and inflammatory responses, and may cause systemic inflammation and insulin resistance., Author Summary Type 2 diabetes is one of the most challenging health problems in the 21st century. Although insulin resistance is regarded as a fundamental defect that precedes the development of type 2 diabetes, the nature and cause of insulin resistance remain unknown. Adipose tissue is an important organ that determines whole-body energy metabolism, and its dysfunction is a critical element in the development of systemic insulin resistance. Adipose mitochondrial function is suppressed in the insulin-resistant state, and increased adipose mitochondrial biogenesis is associated with the reversal of insulin resistance by a PPARγ agonist. However, despite these important observations, little is known about how mitochondrial respiratory dysfunction in white adipose tissue (WAT) causes insulin resistance. To determine whether adipose deficiency of mitochondrial respiratory capacity plays an etiological role in systemic insulin resistance, the metabolic phenotype of mice with mitochondrial OXPHOS (oxidative phosphorylation)–deficient adipose tissue was examined. Crif1 is a protein required for the translation of mtDNA–encoded OXPHOS subunits. Interestingly, mice haploinsufficient for Crif1 in adipose tissue showed reduced OXPHOS capacity and developed marked insulin resistance.

Adipocyte differentiation requires the coordinated activities of several nuclear transcription factors. Recently, mitochondria biogenesis was reported to occur during adipocyte differentiation and following treatment with thiazolidinediones in vitro and in vivo. Crif1 is a translational factor for mitochondrial DNA (mtDNA) and is important for transcription of the mitochondrial oxidative phosphorylation (OXPHOS) complex. To investigate the role of OXPHOS in adipogenesis, we analyzed adipocyte differentiation following disruption of Crif1 in vitro and in vivo. The adipose-specific Crif1 knockout mouse had a lower body weight and less fat mass than wild-type mice. Furthermore, adipocytes were smaller and had a dysplastic morphology in the adipose-specific Crif1 knockout mouse. 3T3-L1 adipocytes or adipose-derived stem cells (ADSCs) that lacked Crif1 expressed lower levels of mtDNA-encoded OXPHOS subunits, and adipocyte differentiation was disrupted. Rosiglitazone treatment did not induce adipogenesis or mitochondria biogenesis in Crif1 knockout ADSCs. These results show that mitochondrial OXPHOS and Crif1 are required for rosiglitazone- and hormone-induced adipogenesis.