Kaempferia parviflora rhizome extract exerts anti-obesity effect in high-fat diet-induced obese C57BL/6N mice.

Kaempferia parviflora (KP) rhizome, also called black ginger, has been used as a herbal medicine for many centuries. This current study was aimed at exploring whether KP rhizome extract (KPE) had anti-obesity effects and the mechanism involved. Five-week-old C57BL/6N male mice were allocated into five groups for 8-week feeding with control diet (CD), high-fat diet (HFD), HFD + 150 mg/kg body weight (BW)/day KPE (HFD+K150), HFD + 300 mg/kg BW/day KPE (HFD+K300), and HFD + 600 mg/kg BW/day KPE (HFD+K600). KPE decreased BW, body fat mass, adipose tissue weight, adipocyte size, and serum levels of glucose, triglycerides, cholesterol, insulin, and leptin in HFD-induced obese C57BL/6N mice. KPE inhibited adipogenesis by decreasing CCAAT/enhancer binding protein α, peroxisome proliferator-activated receptor γ, sterol regulatory element-binding protein-1c, acetyl-CoA carboxylase 1, ATP-citrate lyase, and fatty acid synthase mRNA expression. KPE improved lipolysis by increasing carnitine palmitoyl transferase 1 and hormone-sensitive lipase mRNA expression. These results suggest that KPE may have inhibited HFD-induced obesity by regulating several pathways involved in decreasing adipogenesis and enhancing lipolysis. Thus, the results suggest that KPE (or KP) may be applicable as an anti-obesity agent.

Estimated dietary vitamin D intake and major vitamin D food sources of Koreans: based on the Korea National Health and Nutrition Examination Survey 2016-2019.

BACKGROUND/OBJECTIVES: The purpose of this study was to establish a database (DB) of foods containing vitamin D that were investigated in the Korea National Health and Nutrition Examination Survey (KNHANES), to estimate the dietary vitamin D intake, to evaluate the dietary adequacy of this intake, and to identify the major food sources of Koreans that contain vitamin D. SUBJECTS/METHODS: This study used data from the KNHANES 2016-2019. Individuals aged ≥ 1 year who participated in the nutrition survey (n = 28,418) were included. The dietary intake was assessed by the 24-h dietary recall method and individual dietary vitamin D intake was estimated using a newly established vitamin D DB. Dietary adequacy was evaluated by comparing the dietary intake of the participants with adequate intake (AI) as defined by Dietary Reference Intakes for Koreans (KDRIs) 2020. RESULTS: The average dietary vitamin D intake for all the subjects was 3.13 µg/d, which was 33.1% of AI. Dietary vitamin D intake was lower in rural residents, the elderly, and those with low income. The major food groups that contributed to the total dietary vitamin D intake were fish and shellfish (61.59%), eggs (17.75%), meat (8.03%), milk (4.25%), legumes (3.93%), and grains (3.84%). The top 10 individual food items that contributed to the total vitamin D intake were eggs (17.44%), squid (8.5%), eels (7.44%), salmon (5.35%), mackerel (5.27%), anchovies (4.65%), yellow croakers (4.58%), pork meat (4.47%), soymilk (4.46%), and skipjack tuna (3.80%). CONCLUSION: These results show that the mean dietary vitamin D intake of Koreans is lower than the reference AI level. Nutritional policies need to be put in place to increase the vitamin D intake of Koreans in the future. In addition, comprehensive research on all the sources of vitamin D, including intake of supplements and biosynthesis in the skin, is required.

Dietary zinc intake and sources among Koreans: findings from the Korea National Health and Nutrition Examination Survey 2016-2019.

BACKGROUND/OBJECTIVES: Zinc is an essential trace mineral which is important for the growth and development of the human body and immunological and neurological functions. Inadequate zinc intake may cause zinc deficiency with its adverse consequences. In this study, we aimed to estimate the dietary zinc intake levels and sources among Koreans. SUBJECTS/METHODS: For this secondary analysis, we obtained data from the Korea National Health and Nutrition Examination Survey (KNHANES) 2016-2019. Individuals aged ≥ 1 yr who had completed a 24-h recall were included. The dietary zinc intake of each individual was calculated by applying data from a newly developed zinc content database to the KNHANES raw data. We also compared the extracted data with the sex-, age-specific reference values suggested in the Korean Dietary Reference Intakes 2020. The prevalence of adequate zinc intake was then evaluated by the proportion of the individuals who met the estimated average requirement (EAR). RESULTS: The mean zinc intake of Koreans aged ≥ 1 yr and adults aged ≥ 19 yrs were 10.2 and 10.4 mg/day, equivalent to 147.4% and 140.8% of the EAR, respectively. Approximately 2 in 3 Koreans met the EAR for zinc, but the zinc intake differed slightly among the different age and sex groups. In children aged 1-2 yrs, 2 out of 5 exceeded the upper level of intake, and nearly half of the younger adults (19-29 yrs) and the elders (≥ 75 yrs) did not meet the EAR. The major contributing food groups were grains (38.9%), meats (20.4%), and vegetables (11.1%). The top 5 food contributors to zinc intake were rice, beef, pork, egg, and baechu kimchi, which accounted for half of the dietary intake. CONCLUSION: The mean zinc intake among Koreans was above the recommended level, but 1 in 3 Koreans had inadequate zinc intake and some children were at risk of excessive zinc intake. Our study included zinc intake from diet only, thus to better understand zinc status, further research to include intake from dietary supplements is needed.

Magnesium intake and dietary sources among Koreans: findings from the Korea National Health and Nutrition Examination Survey 2016-2019.

BACKGROUND/OBJECTIVES: Magnesium is an essential nutrient for human health. However, inadequate intake is commonly reported worldwide. Along with reduced consumption of vegetables and fruits and increased consumption of refined or processed foods, inadequate magnesium intake is increasingly reported as a serious problem. This study aimed to assess magnesium intake, its dietary sources, and the adequacy of magnesium intake in Korean populations. SUBJECTS/METHODS: Data was obtained from the Korea National Health and Nutrition Examination Survey 2016-2019 and included individuals aged ≥1 yr who had participated in a nutrition survey (n=28,418). Dietary intake was assessed by 24-h recall, and dietary magnesium intake was estimated using a newly established magnesium database. Diet adequacy was evaluated by comparing dietary intake with the estimated average requirement (EAR) suggested in the Korean Dietary Reference Intakes 2020. RESULTS: The mean dietary magnesium intake of Koreans aged ≥1 yr was 300.4 mg/d, which was equivalent to 119.8% of the EAR. The prevalence of individuals whose magnesium intake met the EAR was 56.8%. Inadequate intake was observed more in females, adolescents and young adults aged 12-29 yrs, elders aged ≥65 yrs, and individuals with low income. About four-fifths of the daily magnesium came from plant-based foods, and the major food groups contributing to magnesium intake were grains (28.3%), vegetables (17.6%), and meats (8.4%). The top 5 individual foods that contributed to magnesium intake were rice, Baechu (Korean cabbage) kimchi, tofu, pork, and milk. However, the contribution of plant foods and individual contributing food items differed slightly by sex and age groups. CONCLUSIONS: This study found that the mean dietary magnesium intake among Koreans was above the recommended intake, whereas nearly one in 2 Koreans had inadequate magnesium intake. To better understand the status of magnesium intake, further research is required, which includes the intake of dietary supplements.

Immunostimulatory activity of hydrolyzed and fermented Platycodon grandiflorum extract occurs via the MAPK and NF-κB signaling pathway in RAW 264.7 cells.

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG) has long been known as a medicinal herb effective in various diseases, including bronchitis and asthma, but is still more widely used for food. Fermentation methods are being applied to increase the pharmacological composition of PG extracts and commercialize them with high added value. This study examines the hydrolyzed and fermented PG extract (HFPGE) fermented with Lactobacillus casei in RAW 264.7 cells, and investigates the effect of amplifying the immune and the probable molecular mechanism. MATERIALS/METHODS: HFPGE's total phenolic, flavonoid, saponin, and platycodin D contents were analyzed by colorimetric analysis or high-performance liquid chromatography. Cell viability was measured by the MTT assay. Phagocytic activity was analyzed by a phagocytosis assay kit, nitric oxide (NO) production by a Griess reagent system, and cytokines by enzyme-linked immunosorbent assay kits. The mRNA expressions of inducible nitric oxide synthase (iNOS) and cytokines were analyzed by reverse transcription-polymerase chain reaction, whereas MAPK and nuclear factor (NF)-κB activation were analyzed by Western blots. RESULTS: Compared to PGE, HFPGE was determined to contain 13.76 times and 6.69 times higher contents of crude saponin and platycodin D, respectively. HFPGE promoted cell proliferation and phagocytosis in RAW 264.7 cells and regulated the NO production and iNOS expression. Treatment with HFPGE also resulted in increased production of interleukin (IL)-1ß, IL-6, tumor necrosis factor (TNF)-α, C-X-C motif chemokine ligand10, granulocyte-colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and monocyte chemoattractant protein-1, and the mRNA expressions of these cytokines. HFPGE also resulted in significantly increasing the phosphorylation of NF-κB p65, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. CONCLUSIONS: Taken together, our results imply that fermentation and hydrolysis result in the extraction of more active ingredients of PG. Furthermore, we determined that HFPGE exerts immunostimulatory activity via the MAPK and NF-κB signaling pathways.

Dietary intake and major source foods of vitamin E among Koreans: findings of the Korea National Health and Nutrition Examination Survey 2016-2019.

BACKGROUND/OBJECTIVES: Vitamin E is essential for health, and although vitamin E deficiency seems rare in humans, studies on estimates of dietary intake are lacking. This study aimed to estimate dietary vitamin E intake, evaluate dietary adequacy of vitamin E, and detail major food sources of vitamin E in the Korean population. SUBJECTS/METHODS: This study used data from the Korea National Health and Nutrition Examination Survey (KNHANES) 2016-2019. Individuals aged ≥ 1 year that participated in a nutrition survey (n = 28,418) were included. Dietary intake was assessed by 24-h recall and individual dietary vitamin E intake was estimated using a newly established vitamin E database. Dietary adequacy was evaluated by comparing dietary intake with adequate intake (AI) as defined by Korean Dietary Reference Intakes 2020. RESULTS: For all study subjects, mean daily total vitamin E intake was 7.00 mg α-tocopherol equivalents, which was 61.6% of AI. The proportion of individuals that consumed vitamin E at above the AI was 12.9%. Inadequate intake was observed more in females, older individuals, rural residents, and those with a low income. Mean daily intakes of tocopherol (α-, ß-, γ-, and δ-forms) and tocotrienol were 6.02, 0.30, 6.19, 1.63, and 1.61 mg, respectively. The major food groups that contributed to total dietary vitamin E intake were grains (22.3%), seasonings (17.0%), vegetables (15.3%), and fish, and shellfish (7.4%). The top 5 individual food items that contributed to total vitamin E intake were baechu kimchi, red pepper powder, eggs, soybean oil, and rice. CONCLUSIONS: This study shows that mean dietary vitamin E intake by Koreans did not meet the reference adequate intake value. To better understand the status of vitamin E intake, further research is needed that considers intake from dietary supplements.

Aqueous extract of Petasites japonicus leaves promotes osteoblast differentiation via up-regulation of Runx2 and Osterix in MC3T3-E1 cells.

BACKGROUND/OBJECTIVES: Petasites japonicus Maxim (P. japonicus) has been used as an edible and medicinal plant and contains many bioactive compounds. The purpose of this study is to investigate the effect of P. japonicus on osteogenesis. MATERIALS/METHODS: The leaves and stems of P. japonicus were separated and extracted with hot water or ethanol, respectively. The total phenolic compound and total polyphenol contents of each extract were measured, and alkaline phosphatase (ALP) activity of each extract was evaluated to determine their effect on bone metabolism. To investigate the effect on osteoblast differentiation of the aqueous extract of P. japonicus leaves (AL), which produced the highest ALP activity among the tested extracts, collagen content was measured using the Sirius Red staining method, mineralization using the Alizarin Red S staining method, and osteocalcin production through enzyme-linked immunosorbent assay analysis. Also, real-time reverse transcription polymerase chain reaction was performed to investigate the mRNA expression levels of Runt-related transcriptional factor 2 (Runx2) and Osterix. RESULTS: Among the 4 P. japonicus extracts, AL had the highest values in all of the following measures: total phenolic compounds, total polyphenols, and ALP activity, which is a major biomarker of osteoblast differentiation. The AL-treated MC3T3-E1 cells showed significant increases in induced osteoblast differentiation, collagen synthesis, mineralization, and osteocalcin production. In addition, mRNA expressions of Runx2 and Osterix, transcription factors that regulate osteoblast differentiation, were significantly increased. CONCLUSIONS: These results suggest that AL can regulate osteoblasts differentiation, at least in part through Runx2 and Osterix. Therefore, it is highly likely that P. japonicus will be useful as an alternate therapeutic for the prevention and treatment of osteoporosis.

Toxicodendron vernicifluum Stokes extract inhibits solid tumor growth and lung metastasis of 4T1 murine mammary carcinoma cells in BALB/c mice.

Toxicodendron vernicifluum Stokes has long been used as a food supplement and traditional herbal medicine in East Asia. We applied a new extraction method to produce Toxicodendron vernicifluum Stokes extract (TVSE), that doesn't contain urushiol (an allergenic toxin) but dose have higher levels of some flavonoids such as fustin and fisetin. This study was conducted to investigate the anticancer effects of TVSE in an in vivo system. Fifty BALB/c mice were acclimated for one week and then injected with 4T1 murine mammary carcinoma cells in mammary fat pads. After 7 days, the mice were randomly divided into 5 groups, and orally administered with 0, 50, 100, 200 or 400 mg of TVSE/kg body weight (BW)/day for 20 days. TVSE reduced tumor volume and weight dose-dependently. The expression of Ki67 was significantly reduced and the number of TUNEL-positive apoptotic cells was significantly increased in the TVSE-treated group over 100 mg/kg BW/day. While tumor nodules were not found in the liver, but only in lungs, the number of tumor nodules was reduced in a dose-dependent manner in the TVSE treated groups compared to the control group. In breast tumors, expression of platelet endothelial cell adhesion molecule (PECAM-1) and vascular endothelial growth factor (VEGF) was reduced by TVSE treatment. TVSE treatment significantly suppressed mRNA expression in tumors of matrix metalloproteinase (MMP)-2, tissue inhibitor of metalloproteinase (TIMP)-1, urokinase-type plasminogen activator (uPA), intercellular adhesion molecule (ICAM)-1, and vascular cell adhesion molecule (VCAM)-1 while increasing plasminogen activator inhibitor (PAI)-1. These results suggest that TVSE is potentially beneficial for the suppression of breast cancer growth and its-associated lung metastasis.

Gynostemma Pentaphyllum Extract Ameliorates High-Fat Diet-Induced Obesity in C57BL/6N Mice by Upregulating SIRT1.

Gynostemma pentaphyllum is widely used in Asia as a herbal medicine to treat type 2 diabetes, dyslipidemia, and inflammation. Here, we investigated the anti-obesity effect and underlying mechanism of G. pentaphyllum extract (GPE) enriched in gypenoside L, gypenoside LI, and ginsenoside Rg3 and obtained using a novel extraction method. Five-week-old male C57BL/6N mice were fed a control diet (CD), high-fat diet (HFD), HFD + 100 mg/kg body weight (BW)/day GPE (GPE 100), HFD + 300 mg/kg BW/day GPE (GPE 300), or HFD + 30 mg/kg BW/day Orlistat (Orlistat 30) for 8 weeks. The HFD-fed mice showed significant increases in body weight, fat mass, white adipose tissue, and adipocyte hypertrophy compared to the CD group; but GPE inhibited those increases. GPE reduced serum levels of triglyceride, total cholesterol, and LDL-cholesterol, without affecting HDL-cholesterol. GPE significantly increased AMPK activation and suppressed adipogenesis by decreasing the mRNA expression of CCAAT/enhancer binding protein-α (C/EBPα), peroxisome proliferator-activated receptor-γ (PPARγ), sterol regulatory element-binding protein-1c (SREBP1c), PPARγ coactivator-1α, fatty acid synthase (FAS), adipocyte protein 2 (AP2), and sirtuin 1 (SIRT1) and by increasing that of carnitine palmitoyltransferase (CPT1) and hormone- sensitive lipase (HSL). This study demonstrated the ameliorative effect of GPE on obesity and elucidated the underlying molecular mechanism.

Cyanidin-3-O-galactoside-enriched Aronia melanocarpa extract attenuates weight gain and adipogenic pathways in high-fat diet-induced obese C57BL/6 mice.

: Aronia melanocarpa are a rich source of anthocyanins that have received considerable interest for their relations to human health. In this study, the anti-adipogenic effect of cyanidin-3-O-galactoside-enriched Aronia melanocarpa extract (AM-Ex) and its underlying mechanisms were investigated in an in vivo system. Five-week-old male C57BL/6N mice were randomly divided into five groups for 8-week feeding with a control diet (CD), a high-fat diet (HFD), or a HFD with 50 (AM-Ex 50), 100 (AM-Ex 100), or 200 AM-Ex (AM-Ex 200) mg/kg body weight/day. HFD-fed mice showed a significant increase in body weight compared to the CD group, and AM-Ex dose-dependently inhibited this weight gain. AM-Ex significantly reduced the food intake and the weight of white fat tissue, including epididymal fat, retroperitoneal fat, mesenteric fat, and inguinal fat. Treatment with AM-Ex (50 to 200 mg/kg) reduced serum levels of leptin, insulin, triglyceride, total cholesterol, and low density lipoprotein (LDL)-cholesterol. Real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed that AM-Ex suppressed adipogenesis by decreasing CCAAT/enhancer binding protein , peroxisome proliferator-activated receptor , sterol regulatory element-binding protein-1c, peroxisome proliferator-activated receptor gamma coactivator-1, acetyl-CoA carboxylase 1, ATP-citrate lyase, fatty acid synthase, and adipocyte protein 2 messenger RNA (mRNA) expressions. These results suggest that AM-Ex is potentially beneficial for the suppression of HFD-induced obesity by modulating multiple pathways associated with adipogenesis and food intake.

Sequencing and Comparative Analysis of the Chloroplast Genome of Angelica polymorpha and the Development of a Novel Indel Marker for Species Identification.

The genus Angelica (Apiaceae) comprises valuable herbal medicines. In this study, we determined the complete chloroplast (CP) genome sequence of A. polymorpha and compared it with that of Ligusticum officinale (GenBank accession no. NC039760). The CP genomes of A. polymorpha and L. officinale were 148,430 and 147,127 bp in length, respectively, with 37.6% GC content. Both CP genomes harbored 113 unique functional genes, including 79 protein-coding, four rRNA, and 30 tRNA genes. Comparative analysis of the two CP genomes revealed conserved genome structure, gene content, and gene order. However, highly variable regions, sufficient to distinguish between A. polymorpha and L. officinale, were identified in hypothetical chloroplast open reading frame1 (ycf1) and ycf2 genic regions. Nucleotide diversity (Pi) analysis indicated that ycf4⁻chloroplast envelope membrane protein (cemA) intergenic region was highly variable between the two species. Phylogenetic analysis revealed that A. polymorpha and L. officinale were well clustered at family Apiaceae. The ycf4-cemA intergenic region in A. polymorpha carried a 418 bp deletion compared with L. officinale. This region was used for the development of a novel indel marker, LYCE, which successfully discriminated between A. polymorpha and L. officinale accessions. Our results provide important taxonomic and phylogenetic information on herbal medicines and facilitate their authentication using the indel marker.

Effect of isoflavone-enriched whole soy milk powder supplementation on bone metabolism in ovariectomized mice.

BACKGROUND/OBJECTIVE: There is intense interest in soy isoflavone as a hormone replacement therapy for the prevention of postmenopausal osteoporosis. A new kind of isoflavone-enriched whole soy milk powder (I-WSM) containing more isoflavones than conventional whole soy milk powder was recently developed. The aim of this study was to investigate the effects of I-WSM on bone metabolism in ovariectomized mice. MATERIALS/METHODS: Sixty female ICR mice individually underwent ovariectomy (OVX) or a sham operation, and were randomized into six groups of 10 animals each as follows: Sham, OVX, OVX with 2% I-WSM diet, OVX with 10% I-WSM diet, OVX with 20% I-WSM diet, and OVX with 20% WSM diet. After an 8-week treatment period, bone mineral density (BMD), calcium, alkaline phosphatase (ALP), tartrate-resistant acid phosphatase (TRAP) 5b, osteocalcin (OC), procollagen 1 N-terminal propeptide (P1NP), and osteoprotegenin (OPG) were analyzed. RESULTS: BMD was significantly lower in the OVX group compared to the Sham group but was significantly higher in OVX + 10% I-WSM and OVX + 20% I-WSM groups compared to the OVX group (P < 0.05). Serum calcium concentration significantly increased in the OVX + 10% and 20% I-WSM groups. Serum ALP levels were significantly lower in the OVX + 10% and 20% I-WSM groups compared to the other experimental groups (P < 0.05). OC was significantly reduced in the OVX group compared to the Sham group (P < 0.05), but a dose-dependent increase was observed in the OVX groups supplemented with I-WSM. P1NP and OPG levels were significantly reduced, while TRAP 5b level was significantly elevated in the OVX group compared with the Sham group, which was not affected by I-WSM (P < 0.05). CONCLUSIONS: This study suggests that I-WSM supplementation in OVX mice has the effect of preventing BMD reduction and promoting bone formation. Therefore, I-WSM can be used as an effective alternative to postmenopausal osteoporosis prevention.

Objectification of the Qi Blood Yin Yang Deficiency Pattern by Using a Facial Color Analysis.

OBJECTIVES: This study aimed to assess a Qi Blood Yin Yang evaluation method systematically and objectively and to identify the correlation between the Qi Blood Yin Yang deficiency pattern (QBYYDP) and facial color. METHODS: Thirty-seven participants (17 males, 20 females) were enrolled in this study. Twenty-four (10 males, 14 females) had ages from 40 to over 60, and 13 (7 males and 6 females) were in their twenties. After sufficient rest, facial images were taken with a camera. Based on the results from a questionnaire survey, we divided the participants into five groups: the normal and the Qi-, Blood-, Yin-, and Yang-deficient groups, after which the relationships between the L, 'a', and 'b' values in the Lab color system and the characteristics of the participants in each of the deficient groups were elucidated using a facial color analysis program. RESULTS: The color analysis for Qi-deficient (QD) participants revealed that the L value was fairly decreased in comparison with the normal participants, but the 'a' and 'b' values were almost the same. A comparison between the normal and the Yang-deficient (YaD) groups revealed that the L values were somewhat lower compared to the normal group, but the 'a' and 'b' values were not statistically different. For the Yin-deficient (YiD) group, the L value was slightly lower compared to the normal group, but the 'a' and 'b' values were almost the same and the R values were slightly increased. For the Blood-deficient (BD) group, the L values were slightly increased compared to the normal group, but the 'a' and 'b' values were decreased slightly. CONCLUSION: This study obtained objective, reliable data for judging the QBYYDP by using facial images and a color analysis program. However, further study with at least 10 or more subjects in each of the deficient groups is necessary to confirm our findings.

Ethanol extract of Innotus obliquus (Chaga mushroom) induces G1 cell cycle arrest in HT-29 human colon cancer cells.

BACKGROUND/OBJECTIVES: Inonotus obliquus (I. obliquus, Chaga mushroom) has long been used as a folk medicine to treat cancer. In the present study, we examined whether or not ethanol extract of I. obliquus (EEIO) inhibits cell cycle progression in HT-29 human colon cancer cells, in addition to its mechanism of action. MATERIALS/METHODS: To examine the effects of Inonotus obliquus on the cell cycle progression and the molecular mechanism in colon cancer cells, HT-29 human colon cancer cells were cultured in the presence of 2.5 - 10 µg/mL of EEIO, and analyzed the cell cycle arrest by flow cytometry and the cell cycle controlling protein expression by Western blotting. RESULTS: Treatment cells with 2.5 - 10 µg/mL of EEIO reduced viable HT-29 cell numbers and DNA synthesis, increased the percentage of cells in G1 phase, decreased protein expression of CDK2, CDK4, and cyclin D1, increased expression of p21, p27, and p53, and inhibited phosphorylation of Rb and E2F1 expression. Among I. obliquus fractions, fraction 2 (fractionated by dichloromethane from EEIO) showed the same effect as EEIO treatment on cell proliferation and cell cycle-related protein levels. CONCLUSIONS: These results demonstrate that fraction 2 is the major fraction that induces G1 arrest and inhibits cell proliferation, suggesting I. obliquus could be used as a natural anti-cancer ingredient in the food and/or pharmaceutical industry.

Effects of pulsed electromagnetic field therapy on delayed-onset muscle soreness in biceps brachii.

OBJECTIVE: to compare the effects of pulsed electromagnetic field (PEMF) therapy and sham treatment on DOMS-related variables in elbow flexors at 24, 48 and 72 h after delayed onset muscle soreness (DOMS) induction exercise. DESIGN: randomized, double-blind, placebo-controlled study. SETTING: Yonsei University laboratory. PARTICIPANTS: In total, 30 healthy male college students. MAIN OUTCOME MEASURES: Muscle soreness, peak torque, median frequency (MDF) and electromechanical delay (EMD) during isometric contraction at 24, 48 and 72 h after DOMS induction exercise. RESULTS: Overall, the application of the PEMF was found to be effective in reducing the physiological deficits associated with DOMS, including improved recovery of perceived muscle soreness, MDF, and EMD during isometric contraction. Our results did not show that PEMF treatment was mechanically more effective for isometric peak torque generation compared to the sham group. CONCLUSION: this study indicates that PEMF may be useful as a modality to reduce DOMS symptoms. However, further well-designed experiments are required to determine optimal treatment dosage and duration, and to investigate the physiological and clinical mechanisms of PEMF on DOMS.

Effect of therapeutic hypothermia according to severity of sepsis in a septic rat model.

AIM OF STUDY: The effects of therapeutic hypothermia (HT) during experimental sepsis may be influenced by disease severity. We experimentally investigated the effect of therapeutic HT on varying disease severity in a septic rat model. MATERIALS AND METHODS: An adult male Sprague-Dawley rat model of intra-abdominal sepsis was used. To modify the disease severity, we used two different models; a moderate severe sepsis model (MSSM) and a severe septic shock model (SSSM). All rats were randomized to a hypothermia group (HT, 30-32°C) or a normothermia group (NT, 36-38°C) 1h after sepsis induction in each model. HT was maintained for 4h and rewarming was conducted for 2h. Survival time was recorded for up to 12h in the SSSM group and 24h in the MSSM group. Acute lung and liver injury, cytokine, and malondialdehyde (MDA) levels were investigated 7h after sepsis induction. Hemodynamic profiles were also evaluated. RESULTS: In the SSSM, there were survival benefits and reduced acute lung and liver injury with therapeutic HT. Therapeutic HT was also associated with significantly reduced levels of plasma interleukin-6 and tissue malondialdehyde (MDA) levels in the liver and lung compared with the NT group in the SSSM. There was a tendency for the mean arterial pressure to be higher in the HT group compared to the NT group in the SSSM. In MSSM, however, there was no such beneficial effect. CONCLUSION: In this rat model of severe septic shock, therapeutic HT showed beneficial effects. In contrast, therapeutic HT did not show protective effect in the moderate sepsis model.

Novel lipolytic enzymes identified from metagenomic library of deep-sea sediment.

Metagenomic library was constructed from a deep-sea sediment sample and screened for lipolytic activity. Open-reading frames of six positive clones showed only 33-58% amino acid identities to the known proteins. One of them was assigned to a new group while others were grouped into Families I and V or EstD Family. By employing a combination of approaches such as removing the signal sequence, coexpression of chaperone genes, and low temperature induction, we obtained five soluble recombinant proteins in Escherichia coli. The purified enzymes had optimum temperatures of 30-35°C and the cold-activity property. Among them, one enzyme showed lipase activity by preferentially hydrolyzing p-nitrophenyl palmitate and p-nitrophenyl stearate and high salt resistance with up to 4 M NaCl. Our research demonstrates the feasibility of developing novel lipolytic enzymes from marine environments by the combination of functional metagenomic approach and protein expression technology.

alpha-Lipoic acid reduces matrix metalloproteinase activity in MDA-MB-231 human breast cancer cells.

alpha-Lipoic acid (LA), a naturally occurring molecule in animal and plant cells, is a potent antioxidant that reportedly exerts beneficial effects on cell proliferation and apoptosis in various cancer cell lines. However, the molecular mechanisms behind the antimetastatic property of LA are not well understood. The present study investigates the effect of LA on metastasis in a cell system. Our hypothesis is that LA inhibits metastasis via inhibition of matrix metalloproteinase (MMP) in vitro. MDA-MB-231 cells, a human breast cancer cell line, were treated with various concentrations of LA (0, 250, 500, or 1000 mumol/L) to measure metastasis, MMP activity, and mRNA expression. The viability of cells was examined by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The effect of LA on metastasis was evaluated using the motility, migration, and invasion assay in vitro. The activity and mRNA expression of MMP-2 and MMP-9 were measured. After LA treatment, cell motility and cell migration were significantly decreased (P < .05). alpha-Lipoic acid also reduced cell invasion through a Matrigel-coated chamber (P < .05). Activities of MMP-2 and MMP-9 were decreased by LA treatment in a dose-dependent manner. RT-PCR analysis confirmed the reduction in mRNA expression level of MMP-2 and MMP-9 by LA treatment. We conclude that in this cell culture model, LA treatment inhibits cancer metastasis, and this inhibition is likely due to the decrease in the activity and mRNA expression levels of MMP-2 and MMP-9 caused by LA.

Cloning and characterization of a new cold-active lipase from a deep-sea sediment metagenome.

To search for new cold-active lipases, a metagenomic library was constructed using cold-sea sediment samples at Edison Seamount and was screened for lipolytic activities by plating on a tricaprylin medium. Subsequently, a fosmid clone was selected, and the whole sequence of 36 kb insert of the fosmid clone was determined by shotgun sequencing. The sequence analysis revealed the presence of 25 open reading frames (ORF), and ORF20 (EML1) showed similarities to lipases. Phylogenetic analysis of EML1 suggested that the protein belonged to a new family of esterase/lipase together with LipG. The EML1 gene was expressed in Escherichia coli, and purified by metal-chelating chromatography. The optimum activity of the purified EML1 (rEML1) occurred at pH 8.0 and 25 degrees C, respectively, and rEML1 displayed more than 50% activity at 5 degrees C. The activation energy for the hydrolysis of olive oil was determined to be 3.28 kcal/mol, indicating that EML1 is a cold-active lipase. rEML1 preferentially hydrolyzed triacylglycerols acyl-group chains with long chain lengths of > or = 8 carbon atoms and displayed hydrolyzing activities toward various natural oil substrates. rEML1 was resistant to various detergents such as Triton X-100 and Tween 80. This study represents an example which developed a new cold-active lipase from a deep-sea sediment metagenome.

L-carnitine stimulates lipolysis via induction of the lipolytic gene expression and suppression of the adipogenic gene expression in 3T3-L1 adipocytes.

We investigated the lipolytic effects of L-carnitine in 3T3-L1 adipocytes. L-carnitine at 10-S100 nM suppressed lipid accumulation. The release of glycerol and free fatty acid into the medium was significantly increased by 1.5- and 1.7- fold, respectively, by the addition of 100 nM L-carnitine compared with the control (P < .05). The mRNA levels of hormone-sensitive lipase, carnitine palmitoyltransferase I-a, and acyl-coenzyme A oxidase, all of which participate in lipid catabolism, were increased in the presence of 100 nM L-carnitine by 2.8-, 2.2-, and 1.6-fold, respectively (P < .05). However, the expression of peroxisome proliferator-activated receptor-gamma and adipose-specific fatty acid-binding protein, which are involved in adipogenesis, were down-regulated by L-carnitine in 3T3-L1 adipocytes (P < .05). These results suggest an anti-obesity action of L-carnitine. L-carnitine may modulate lipid metabolism by stimulation of lipolysis and beta-oxidation accompanied by corresponding changes in gene expression and suppression of adipogenic gene expression.