Cell-free culture supernatant of Lactobacillus curvatus Wikim38 inhibits RANKL-induced osteoclast differentiation and ameliorates bone loss in ovariectomized mice.

This study was conducted to investigate the inhibitory effects of the cell-free culture supernatant of Lactobacillus curvatus Wikim 38 (LC38-CS) on RANKL-induced osteoclast differentiation and bone loss in a mice model of ovariectomy-induced post-menopausal osteoporosis. LC38-CS inhibited the RANKL-induced differentiation of bone marrow-derived macrophages (BMDMs) into osteoclasts in a dose-dependent manner. F-actin ring formation and bone resorption were also reduced by LC38-CS treatment of RANKL-treated BMDMs. In addition, LC38-CS decreased the RANKL-induced activation of the TRAF6/NF-κB/MAPKs axis at the early stage and the expression of osteoclastogenesis-related genes in BMDMs treated with RANKL. PRMT1 and ADMA levels, new biomarkers for osteoclastogenesis, were decreased by LC38-CS treatment. The administration of LC38-CS increased bone volume and bone mineral density in ovariectomized mice in µ-CT analysis. These findings suggest that LC38-CS inhibited RANKL-induced osteoclast differentiation by the downregulation of molecular mechanisms and exerted anti-osteoporotic effects.

Effects of dietary Achyranthes japonica extract supplementation on the growth performance, total tract digestibility, cecal microflora, excreta noxious gas emission, and meat quality of broiler chickens.

The present study was investigated the effects of dietary Achyranthes japonica extract (AJE) supplementation on the growth performance, total tract digestibility, cecal microflora, excreta noxious gas emission, breast meat quality, and organ weight in broiler chickens. In total, 640 Ross × Ross male broiler chickens (1-day-old) were randomly distributed into 4 dietary treatments with 10 replicate cages (16 birds/replicate) per treatment group for 5 wk. The dietary treatments included a control basal diet without AJE, and diets with 0.025, 0.05, or 0.1% AJE. Body weight gain, feed intake, and feed conversion improved linearly with the supplementation of AJE over the experimental period (days 1 to 35) (P < 0.05). Dietary AJE supplementation caused a significant increase in the apparent total tract digestibility of dry matter and nitrogen (linear, P < 0.05). The cecal Lactobacillus, E. coli, and Salmonella counts were linearly affected with increasing dietary AJE supplementation (P < 0.05). With increasing levels of AJE, excreta ammonia gas concentration showed a linear decrease (P < 0.05). The breast muscle weight linearly increased, along with a decrease in the abdominal fat weight, in treatment groups fed with AJE (P < 0.05). These results indicate that dietary addition with increasing AJE linearly improved growth performance, total tract digestibility, cecal microflora, excreta ammonia gas emission, and abdominal fat weight in broiler chickens.

The effects of betaine supplementation in diets containing different levels of crude protein and methionine on the growth performance, blood components, total tract nutrient digestibility, excreta noxious gas emission, and meat quality of the broiler chickens.

The effects of betaine supplementation on growth performance, blood components, nutrient digestibility, excreta noxious gas emission, and meat quality of broiler chickens were examined using different dietary crude protein (CP) and methionine (Met) levels. A total of 768 Ross 308 broiler chickens were allotted to four treatments, with 12 replications of each treatment conducted over 6 wk. Treatments were factorially designed, with 2 levels of CP [Starter: CP 21% (low Met) and 23% (high Met); Finisher: CP 18% (low Met) and 20% (high Met)] and 2 levels of betaine supplementation (0 and 0.12%). Body weight gain and feed conversion improved significantly as dietary levels of protein increased (P < 0.05), but the results for betaine supplementation differed. The concentrations of serum total protein, albumin, and glutathione peroxidase (GPx) were elevated by either the supplementary betaine or the CP (P < 0.05). In addition, serum albumin concentration significantly increased in groups fed low CP amounts and betaine 0.12% compared with groups fed low CP only (P < 0.05). Total tract digestibility of nitrogen in broilers fed high CP amounts or 0.12% betaine, was observed to be greater than that in groups fed low CP amounts or no betaine treatment (P < 0.05). Supplemental betaine affected excreta ammonia gas emission, and hydrogen sulfide concentrations decreased significantly in low CP-fed groups (P < 0.05). Breast meat quality and relative organ weights were not influenced by CP levels or dietary betaine supplementation. These results suggest that betaine does not increase productivity, but may affect serum total protein, albumin, GPx, excreta ammonia emission, and nitrogen digestibility in broiler chickens. In addition, betaine supplementation is more effective in increasing serum albumin concentration when it was added in low CP (low Met) diets.

A novel tumor-based epithelial-to-mesenchymal transition score that associates with prognosis and metastasis in patients with Stage II/III colorectal cancer.

It is increasingly appreciated that host factors within the tumor center and microenvironment play a key role in dictating colorectal cancer (CRC) outcomes. As a result, the metastatic process has now been defined as a result of epithelial-mesenchymal transition (EMT). Establishment of the role of EMT within the tumor center and its effect on the tumor microenvironment would be beneficial for prognosis and therapeutic intervention in CRC. The present study assessed five immunohistochemical EMT markers within the tumor center on a 185 Stage II/III CRC patient tissue microarray. In 185 patients with CRC, cytoplasmic snail (HR 1.94 95% confidence interval [CI] 1.15-3.29, p = 0.012) and a novel combined EMT score (HR 3.86 95% CI 2.17-6.86, p < 0.001) were associated with decreased cancer-specific survival. The combined EMT score was also associated with increased tumor budding (p = 0.046), and systemic inflammation (p = 0.007), as well as decreased memory T-cells within the stroma (p = 0.030) and at the invasive margin (p = 0.035). Furthermore, the combined EMT score was associated with cancer-specific survival independent of TNM-stage (HR 4.12 95% CI 2.30-7.39, p < 0.001). In conclusion, a novel combined EMT score stratifies patient's survival in Stage II/III CRC and associates with key factors of tumor metastasis. Therefore, the combined EMT score could be used to identify patients at risk of micrometastases and who may benefit from standard adjuvant therapy, potentially in combination with EMT blockade.

Effects of a protease and essential oils on growth performance, blood cell profiles, nutrient retention, ileal microbiota, excreta gas emission, and breast meat quality in broiler chicks.

A total of 360 Ross male broiler chicks (39.8 ± 1.8 g) were used in a five week experiment to determine the effect of a protease and essential oils (EO) on growth performance, blood cell profile, nutrient retention, ileal microbiota, excreta gas emission, and breast meat quality in broiler chicks. Broiler chicks were randomly allotted to four dietary treatments with 15 birds/cage and six cages/treatment. Experimental treatments were arranged as a 2 × 2 factorial with two levels of protease (0 and 0.02%) and two levels of EO (0 and 0.03%). For days 8 to 21 and overall, body weight gain and the feed conversion ratio were better in broilers fed diets supplemented with protease (P < 0.05) than in those fed diets without protease supplementation. Protease and/or EO increased (P < 0.05) the total tract retention of dry matter, nitrogen, or gross energy, and decreased the excreta ammonia gas emission. In addition, there was a significant interaction between the protease and EO on total tract retention of nitrogen and excreta ammonia gas emission (P < 0.05). The density of ileal Lactobacillus increased and Escherichia coli decreased in broilers (P < 0.05) by the addition of EO to the diet. There were no significant differences in the measurements of breast meat quality and organ weight of broilers fed diets with protease or EO. In conclusion, diets with a combination of a protease and EO improved total tract retention of nitrogen and excreta ammonia gas emission in growing broiler chicks.

Effect of dietary Spirulina (Arthrospira) platensis on the growth performance, antioxidant enzyme activity, nutrient digestibility, cecal microflora, excreta noxious gas emission, and breast meat quality of broiler chickens.

This study examined the effects of dietary Spirulina (Arthrospira) platensis supplementation on growth performance, antioxidant enzyme activity, nutrient digestibility, cecal microflora, excreta noxious gas emission, organ weight and breast meat quality in broiler chickens. In total, 800 Ross 308 male broiler chickens (1-d-old) were randomly divided into 5 dietary treatments with 10 replicate cages (16 birds/replicate) per treatment for 5 wk. The dietary treatments were a control basal diet without Spirulina or with 0.25, 0.5, 0.75, or 1.0% Spirulina. Body weight gain, feed conversion, and/or European production efficiency index improved linearly with supplementation of Spirulina during d 8 to 21, 22 to 35, and overall d 1 to 35 (P < 0.05). Dietary Spirulina supplementation caused a significant increase in the serum enzyme activity of superoxide dismutase and glutathione peroxidase (linear, P < 0.05). Apparent total tract digestibility of dry matter and nitrogen showed a linear increase in Spirulina supplementation (P < 0.05). Cecal Lactobacillus count linearly increased and excreta ammonia gas emission linearly decreased, as dietary Spirulina supplementation increased (P < 0.05). There were no significant effects on relative organ weight and breast meat quality of broilers fed with Spirulina diets; however, 7 d drip loss linearly decreased in treatment groups fed with Spirulina (P < 0.05). These results indicate that adding Spirulina to the diet of broilers can improve antioxidant enzyme activity, dry matter and nitrogen digestibility, cecal Lactobacillus population, excreta ammonia gas emission, and 7 d drip loss of breast meat. In addition, dietary inclusion of 1.0% Spirulina powder might provide a good alternative to improve broiler chicken production.

Effects of dietary methyl sulfonyl methane (MSM) supplementation on growth performance, nutrient digestibility, meat quality, excreta microbiota, excreta gas emission, and blood profiles in broilers.

A 29-d trial was conducted to evaluate the effects of dietary methyl sulfonyl methane (MSM) supplementation on growth performance, meat quality, nutrient digestibility, excreta microbiota, excreta gas emission, and blood profiles in broilers. A total of 816 1-day-old male Ross 308 broilers (44 ± 0.44 g) were assigned to 4 dietary treatments, composed of 12 replicates with 17 birds per replicate. The 4 treatments were: 1) CON, basal diet; 2) S1, CON + 0.05% MSM; 3) S2, CON + 0.10% MSM; 4) S3, CON + 0.20% MSM. In the current study, body weight (BW) on d 14 and 29 showed significant improvement as dietary MSM increased from 0.05% to 0.20% (P < 0.05). During d 1 to 14 and overall, higher (P < 0.05) body weight gain (BWG) and lower feed conversion ratio (FCR) were observed in broilers fed MSM diets. Between d 15 and 29, higher (P < 0.05) BWG was observed in broilers fed MSM diets. Redness (a*) was increased linearly (P < 0.05) in broilers fed MSM diets. On d 3, 5, and 7, drip loss was decreased linearly (P < 0.05) in broilers fed MSM diets. Lactobacillus and E. coli were effected linearly (P < 0.05) in broilers fed MSM diets. Alanine aminotransferase (ALT), white blood cells (WBC) and lymphocytes were improved linearly (P < 0.05) in broilers fed MSM diets. In conclusion, dietary supplementation MSM has positive effects on growth performance, meat quality, excreta microbiota, and blood profiles in broilers.

Efficacy of 1,3-diacylglycerol as a fat emulsifier in low-density diet for broilers.

An experiment was conducted to study the effects of dietary supplementation of 1,3-diacylglycerol (DAG) as an emulsifier in a low-density diet on growth performance, nutrient digestibility, blood characteristics, and meat quality in broilers. A total of 480 1-day-old male 308 Ross broiler chicks were randomly allocated into 1 of the following 5 treatments: Control, fed a basal diet (CON), basal diet minus 100 kcal ME diet (LE), T1 (LE + 0.075% 1,3-DAG), T2 (LE + 0.10% 1,3-DAG), and T3 (LE + 0.15% 1,3-DAG).The supplementation of low energy diet to broilers reduced (P < 0.05) feed intake compared to CON diet during wk 1 of the experiment. The supplementation of LE diet with 0.075, 0.10, and 0.15% 1,3-DAG linearly increased (P = 0.09) body weight gain (BWG) and decreased (P = 0.08) feed intake (FI) during wk 1. During wk 2 to 3 and overall (0 to 5 wk), there was linear increase (P < 0.05) in BWG and decrease in feed conversion ratio (FCR) in LE diet supplemented with graded levels of emulsifier. The LE diet reduced (P < 0.05) dry matter (DM) and energy digestibility (P = 0.07) compared with CON diet. Supplementing energy-reduced diet with different levels of 1,3-DAG linearly increased (P < 0.05) DM, and energy digestibility, but no significant differences were observed in blood profiles, meat quality (except drip loss) of broilers.In conclusion, DAG positively affected growth performances and nutrient digestibility in broilers. However, meat quality and serum profiles were unaffected in broilers fed an energy-reduced diet supplemented with DAG.

Epac2a-null mice exhibit obesity-prone nature more susceptible to leptin resistance.

BACKGROUND: The exchange protein directly activated by cAMP (Epac), which is primarily involved in cAMP signaling, has been known to be essential for controlling body energy metabolism. Epac has two isoforms: Epac1 and Epac2. The function of Epac1 on obesity was unveiled using Epac1 knockout (KO) mice. However, the role of Epac2 in obesity remains unclear. METHODS: To evaluate the role of Epac2 in obesity, we used Epac2a KO mice, which is dominantly expressed in neurons and endocrine tissues. Physiological factors related to obesity were analyzed: body weight, fat mass, food intake, plasma leptin and adiponectin levels, energy expenditure, glucose tolerance, and insulin and leptin resistance. To determine the mechanism of Epac2a, mice received exogenous leptin and then hypothalamic leptin signaling was analyzed. RESULTS: Epac2a KO mice appeared to have normal glucose tolerance and insulin sensitivity until 12 weeks of age, but an early onset increase of plasma leptin levels and decrease of plasma adiponectin levels compared with wild-type mice. Acute leptin injection revealed impaired hypothalamic leptin signaling in KO mice. Consistently, KO mice fed a high-fat diet (HFD) were significantly obese, presenting greater food intake and lower energy expenditure. HFD-fed KO mice were also characterized by greater impairment of hypothalamic leptin signaling and by weaker leptin-induced decrease in food consumption compared with HFD-fed wild-type mice. In wild-type mice, acute exogenous leptin injection or chronic HFD feeding tended to induce hypothalamic Epac2a expression. CONCLUSIONS: Considering that HFD is an inducer of hypothalamic leptin resistance and that Epac2a functions in pancreatic beta cells during demands of greater work load, hypothalamic Epac2a may have a role in facilitating leptin signaling, at least in response to higher metabolic demands. Thus, our data indicate that Epac2a is critical for preventing obesity and thus Epac2a activators may be used to manage obesity and obesity-mediated metabolic disorders.

Evaluation of phosphorus adsorption capacity of sesame straw biochar on aqueous solution: influence of activation methods and pyrolysis temperatures.

The phosphorus (P) adsorption characteristic of sesame straw biochar prepared with different activation agents and pyrolysis temperatures was evaluated. Between 0.109 and 0.300 mg L(-1) in the form of inorganic phosphate was released from raw sesame straw biochar in the first 1 h. The release of phosphate was significantly enhanced from 62.6 to 168.2 mg g(-1) as the pyrolysis temperature increased. Therefore, sesame straw biochar cannot be used as an adsorbent for P removal without change in the physicochemical characteristics. To increase the P adsorption of biochar in aqueous solution, various activation agents and pyrolysis temperatures were applied. The amount of P adsorbed from aqueous solution by biochar activated using different activation agents appeared in the order ZnCl2 (9.675 mg g(-1)) > MgO (8.669 mg g(-1)) ⋙ 0.1N-HCl > 0.1N-H2SO4 > K2SO4 ≥ KOH ≥ 0.1N-H3PO4, showing ZnCl2 to be the optimum activation agent. Higher P was adsorbed by the biochar activated using ZnCl2 under different pyrolysis temperatures in the order 600 °C > 500 °C > 400 °C > 300 °C. Finally, the amount of adsorbed P by activated biochar at different ratios of biochar to ZnCl2 appeared in the order 1:3 ≒ 1:1 > 3:1. As a result, the optimum ratio of biochar to ZnCl2 and pyrolysis temperature were found to be 1:1 and 600 °C for P adsorption, respectively. The maximum P adsorption capacity by activated biochar using ZnCl2 (15,460 mg kg(-1)) was higher than that of typical biochar, as determined by the Langmuir adsorption isotherm. Therefore, the ZnCl2 activation of sesame straw biochar was suitable for the preparation of activated biochar for P adsorption.

Antioxidant Enzyme Activity and Meat Quality of Meat Type Ducks Fed with Dried Oregano (Origanum vulgare L.) Powder.

One-day-old Cherry valley meat-strain ducks were used to investigate the effect of supplemental dried oregano powder (DOP) in feed on the productivity, antioxidant enzyme activity, and breast meat quality. One hundred sixty five ducks were assigned to 5 dietary treatments for 42 days. The dietary treatment groups were control group (CON; no antibiotic, no DOP), antibiotic group (ANT; CON+0.1% Patrol), 0.1% DOP (CON+0.1% DOP), 0.5% DOP (CON+0.5% DOP), and 1.0% DOP (CON+1.0% DOP). Upon feeding, 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity of oregano extracts was higher than that of tocopherol, although it was less than that of ascorbic acid. As a result of in vivo study, DOP in the diet showed no effects on final body weight, feed intake, or feed conversion ratio. However, dietary 0.5% and 1% DOP supplementation caused a significant increase in the serum enzyme activity of superoxide dismutase (SOD) compared with CON and ANT, while glutathione peroxidase (GPx) in tissue was increased as compared to ANT (p<0.05). Cooking loss from ducks fed with DOP decreased compared with the control ducks. Thiobarbituric acid reactive substance (TBARS) values of duck breast meat at 5 d post slaughter was found to be significantly reduced in ducks whose diets were supplemented with 0.5% and 1% DOP (p<0.05). These results suggest that diets containing 0.5% and 1% DOP may beneficially affect antioxidant enzyme activity of GPx and SOD, improve meat cooking loss, and reduce TBARS values in breast meat at 5 d of storage in ducks.

First Report of Powdery Mildew Caused by Golovinomyces biocellatus on Agastache rugosa in Korea.

Agastache rugosa (Fisch. & C.A. Mey.) Kuntze, known as Korean mint, is an aromatic plant in the Lamiaceae. It is widely distributed in East Asian countries and is used as a Chinese traditional medicine. In Korea, fresh leaves are commonly added to fish soups and stews (3). In November 2008, several dozen Korean mints plants growing outdoors in Gimhae City, Korea, were found to be severely infected with a powdery mildew. The same symptoms had been observed in Korean mint plots in Busan and Miryang cities from 2008 to 2013. Symptoms first appeared as thin white colonies, which subsequently developed into abundant hyphal growth on stems and both sides of the leaves. Severe disease pressure caused withering and senescence of the leaves. Voucher specimens (n = 5) were deposited in the Korea University Herbarium (KUS). Appressoria on the mycelium were nipple-shaped or nearly absent. Conidiophores were 105 to 188 × 10 to 13 µm and produced 2 to 4 immature conidia in chains with a sinuate outline, followed by 2 to 3 cells. Foot-cells of the conidiophores were straight, cylindrical, slightly constricted at the base, and 37 to 58 µm long. Conidia were hyaline, ellipsoid to barrel-shaped, measured 25 to 40 × 15 to 23 µm (length/width ratio = 1.4 to 2.1), lacked distinct fibrosin bodies, and showed reticulate wrinkling of the outer walls. Primary conidia were obconically rounded at the apex and subtruncate at the base. Germ tubes were produced at the perihilar position of conidia. No chasmothecia were observed. The structures described above were typical of the Oidium subgenus Reticuloidium anamorph of the genus Golovinomyces. The measurements and morphological characteristics were compatible with those of G. biocellatus (Ehrenb.) V.P. Heluta (1). To confirm the identification, molecular analysis of the sequence of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) of isolate KUS-F27200 was conducted. The complete ITS rDNA sequence was amplified using primers ITS5 and P3 (4). The resulting 514-bp sequence was deposited in GenBank (Accession No. KJ585415). A GenBank BLAST search of the Korean isolate sequence showed >99% similarity with the ITS sequence of many G. biocellatus isolates on plants in the Lamiaceae (e.g., Accession Nos. AB307669, AB769437, and JQ340358). Pathogenicity was confirmed by gently pressing diseased leaf onto leaves of five healthy, potted Korean mint plants. Five non-inoculated plants served as a control treatment. Inoculated plants developed symptoms after 7 days, whereas the control plants remained symptomless. The fungus present on inoculated plants was identical morphologically to that observed on the original diseased plants. The pathogenicity test was repeated with identical results. A powdery mildew on A. rugosa caused by G. biocellatus was reported from Romania (2). To our knowledge, this is the first report of powdery mildew caused by G. biocellatus on A. rugosa in Korea. The plant is mostly grown using organic farming methods with limited chemical control options. Therefore, alternative control measures should be considered. References: (1) U. Braun and R. T. A. Cook. Taxonomic Manual of the Erysiphales (Powdery Mildews), CBS Biodiversity Series No. 11. CBS, Utrecht, 2012. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., online publication, USDA ARS, retrieved 17 February 2014. (3) T. H. Kim et al. J. Sci. Food Agric. 81:569, 2001. (4) S. Takamatsu et al. Mycol. Res. 113:117, 2009.

First Report of Leaf Spot Caused by a Phoma sp. on Schisandra chinensis in Korea.

Schisandra chinensis (Turcz.) Baill. is a deciduous woody vine native to northern China and the Russian Far East. Its berries have long been used in traditional Asian medicine. In Korea, S. chinensis is one of 10 major medicinal crops and, as of 2011, the production is 6,892 metric tons from 1,749 ha of cultivation area (1). During summer to autumn of 2011 and 2012, leaf spots were observed on S. chinensis (cv. Cheongsun) with disease incidence of 100% in many locations of Jangsu County, Korea. Early symptoms appeared as small, circular, and pale brown spots. Each spot increased in size, became grayish brown and necrotic, and finally developed concentric rings with a definite margin. Some spots coalesced to cover nearly half of the leaves, often becoming torn and giving a shot hole effect. The infected leaf tissue contained blackish pycnidia from which masses of conidia were released in a humid environment. The pycnidia were brown, globose to pyriform, ostiolate, and 45 to 160 µm in diameter. Conidia were hyaline, smooth, oval to ellipsoidal, aseptate or medianly 1-septate, very occasionally 2-septate, slightly constricted at the septa, 4 to 11 × 2.5 to 5 µm, and contained small oil drops. These morphological characteristics were consistent with the generic concept of Phoma (2). Three monoconidial isolates were successfully cultured by diluting conidia mass in sterile water and streaking conidia suspension on potato dextrose agar (PDA). A representative isolate was deposited in the Korean Agricultural Culture Collection (Accession No. KACC47113) and used for pathogenicity test and molecular analysis. Inoculum for a pathogenicity test was prepared by harvesting conidia from 30-day-old cultures (12-h diurnal cycle, 25°C) and a conidial suspension in water (1.1 × 107 conidia/ml) was sprayed onto leaves of three healthy seedlings (cv. Cheongsun). Three seedlings serving as controls were sprayed until runoff with sterile distilled water. The plants were separately covered with plastic bags for 48 h in a glasshouse. After 10 days, typical leaf spot symptoms developed on the leaves inoculated with the fungus. Phoma sp. was re-isolated from those lesions, confirming Koch's postulates. No symptoms were observed on controls. The pathogenicity test was conducted twice. Fungal DNA was extracted, and the complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced directly. The resulting 520-bp sequence was deposited in GenBank (Accession No. KC928322). The sequence showed over 99% similarity with many Phoma species from various substrates, but no exact matches. Phoma leaf spot of S. chinensis was once recorded in Korea without pathogenicity test and culture deposition (3). Phoma glomerata was recorded as a causal fungus of leaf spot disease on S. chinensis in China (4). The Korean isolates differ from P. glomerata in having larger conidia and are separated from it in ITS sequence data. Therefore, we tentatively place the Korean isolates as unidentified Phoma sp. To our knowledge, this is the first confirmed report of leaf spot disease caused by a Phoma sp. in Korea. References: (1) Anonymous. Statistics of Cultivation and Production of Industrial Crops in 2011. Korean Ministry for Food, Agriculture, Forestry and Fisheries. 2012. (2) M. M. Aveskamp et al. Mycologia 101:363, 2009. (3) E. J. Lee et al. Compendium of Medicinal Plant Diseases with Color Plates. Nat. Inst. Agric. Sci., Suwon, Korea. 1991. (4) X. Wang et al. Plant Dis. 96:289, 2012.

Antioxidant Activities of Achyranthes japonica Nakai Extract and Its Application to the Pork Sausages.

Influence of Achyranthes japonica Nakai Extract (AJNE) on properties of pork sausages were studied in the present investigation. AJNE was added to sausages alone or in combination with ascorbic acid to obtain a comparative analysis on properties of control and ascorbic acid added-sausages. Results showed that addition of 0.05% AJNE led to a decrease in color L* and whiteness (W), and an increase in color b* of pork sausage samples (p<0.05). Although color a* of pork sausages containing AJNE was not significantly different, ascorbic acid added-sausages were highest amongst other treatments (p<0.05). Sausages containing AJNE had lower non-heme iron values and peroxide value (POV) than control sausages (p<0.05); however, high nitrosomyoglobin content was observed in AJNE added-sausages (p<0.05). Ascorbic acid led to a decrease in residual nitrite concentration of sausages (p<0.05), but no difference was found in AJNE added-sausages. Free radical scavenging analysis showed that AJNE did not affect 1,1-diphenyl -2-picrylhydrazyl (DPPH) activity of sausages, whereas ascorbic acid added-sausages showed relatively higher activity among the samples (p<0.05). Addition of AJNE had no influence on texture properties of sausages. In sensory evaluation, AJNE treatment had significant effects on color (p<0.05), but no significant effects on aroma, flavor, springiness, juiciness, and overall acceptability. In conclusion, the addition of AJNE, as a natural supplement may offer natural antioxidants for pork sausages, and appears to be particularly effective in inducing changes in non-heme iron concentration, POV value and nitrosomyglobin content.

The effects of cranio-cervical flexion on activation of swallowing-related muscles.

We tested the effects of cranio-cervical flexion (CCF) on activation of swallowing-related muscles while swallowing liquid in a sample of 45 healthy volunteers. Activation following CCF movement was examined across two positions (supine and sitting) and, three pressure levels and two different postures were examined in each condition, respectively. When CCF was applied, activation of swallowing-related muscles was significantly increased compared to the neutral neck position, and such findings were found across both the supine and sitting positions. Also in the supine position, when the pressure level of the stabilizer was escalated, there was a significant difference in the activity of the swallowing-related muscles compared to the baseline level. In conclusion, our results suggest that CCF may be a viable method to enhance the effectiveness of swallowing-related muscles by changing neck position. When CCF is applied, the stability of the deep flexor muscles must be secured first after which superficially located muscles may better assist swallowing with less effort.

Effects of dietary combination of n-3 and n-9 fatty acids on the deposition of linoleic and arachidonic acid in broiler chicken meats.

To minimize the amount of n-6 fatty acids in broiler chicken meat, 120 Cobb × Ross male broilers were divided into 6 different groups and fed a basal corn-soybean meal diet containing 5% fat from 5 different lipid sources: 1) a commercial mix of animal and vegetable oil, 2) soybean oil and olive oil (2.5% each), 3) flaxseed oil and olive oil (2.5% each), 4) flaxseed oil, eicosapentaenoic acid (C20:5; EPA; n-3), and olive oil (2.45, 0.05, and 2.5% respectively; FEO), 5) flaxseed oil, docosahexaenoic acid (C22:6; DHA; n-3), and olive oil (2.45, 0.05, and 2.5% respectively; FDO), and 6) fish oil and olive oil (2.5% each; FHO). At 6 and 9 wk, one bird per pen (4 pens per treatment) was processed, and liver, breast, and thigh samples were collected and used for fatty acid profiles or Δ6- and Δ9-desaturase mRNA gene expression levels. The deposition of linoleic acid (C18:2; n-6) or arachidonic acid (C20:4; n-6) was decreased in breast and thigh muscles of chickens fed n-3 fatty acids for 9 wk compared with chickens fed animal and vegetable oil and soybean oil and olive oil diets (P < 0.05). The addition of EPA to the diet (FEO; P > 0.05) did not reduce the deposition of linoleic acid and arachidonic acid as much as DHA (FDO; P < 0.05), and it suppressed the expression of Δ6- and Δ9-desaturase. When EPA and DHA were blended (FHO) and supplied to broiler chickens for 9 wk, EPA and DHA combination effects were observed on the deposition of LA and arachidonic acid in breast and thigh muscles. Thereby, the addition of a mixed EPA and DHA to a broiler chicken diet may be recommendable to reduce arachidonic acid accumulation in both broiler chicken breast and thigh meats, providing a functional broiler chicken meat to consumers.

First Report of Powdery Mildew Caused by Erysiphe heraclei on Dill in Korea.

Dill (Anethum graveolens L.) is a scented herb belonging to the family Apiaceae. The plant has a long and ancient history in many countries as a culinary and medicinal herb. In October 2008, plants showing typical symptoms of powdery mildew disease were found in polythene tunnels in Icheon, Korea. Symptoms first appeared as thin white colonies, which subsequently showed abundant growth on the leaves and stems. Most diseased plantings were unmarketable and shriveled without being harvested. The damage caused by powdery mildew infections on dill has reappeared every year, with confirmation of the causal agent made again in 2011. Voucher specimens were deposited in the Korea University Herbarium (KUS). Hyphae were septate, branched, and 4 to 7 µm wide. Appressoria on the mycelium were multilobed or moderately lobed. Conidiophores were unbranched, cylindrical, 80 to 140 × 8 to 10 µm, straight or slightly flexuous in foot cells, and produced conidia singly, followed by two to three cells. Conidia were oblong elliptical to oblong, 28 to 50 × 14 to 18 µm, lacked fibrosin bodies, and produced germ tubes on the subterminal position, with angular/rectangular wrinkling of the outer walls. Primary conidia were apically conical, basally subtruncate, and generally smaller than the secondary conidia. No chasmothecia were found but the above characteristics are consistent with Erysiphe heraclei DC. (1). To confirm the identity of the causal fungus, the complete internal transcribed spacer (ITS) region of rDNA from isolate KUS-F26425 was amplified with primers ITS5 and P3 as described by Takamatsu et al. (3) and directly sequenced. The resulting 630-bp sequence was deposited in GenBank (Accession No. JQ517297). Comparison with the sequences available in the GenBank database revealed that the isolate showed >99% sequence similarity with those of E. heraclei from Pleurospermum camtschaticum (GU173850) and Daucus carota (EU371725). Pathogenicity was confirmed through inoculation by gently pressing diseased leaves onto leaves of five healthy potted dill plants. Five noninoculated plants served as controls. Plants were maintained in a greenhouse at 22 ± 2°C. Inoculated plants developed signs and symptoms after 7 days, whereas the control plants remained healthy. The fungus present on the inoculated plants was morphologically identical to that originally observed on diseased plants. Powdery mildew caused by E. heraclei on dill has been known worldwide where the plant is cultivated (2). In East Asia, however, dill powdery mildew was known only from Taiwan (4). To our knowledge, this is the first report of powdery mildew infections by E. heraclei on dill in Korea. Since cultivation of dill was only recently started on a commercial scale in Korea, powdery mildew infections pose a serious threat to safe production of this herb, especially in organic farming where chemical control would be prohibited. References: (1) U. Braun. Beih. Nova Hedw. 89:1, 1987. (2) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ January 28, 2012. (3) S. Takamatsu et al. Mycol. Res. 113:117, 2009. (4) J. G. Tsay. Trans. Mycol. Soc. Repub. China 5:1, 1990.

Determination of tadalafil and N-desmethylsibutramine in health and dietary supplements using ultra-performance liquid chromatography (UPLC) coupled with quadrupole-time-of-flight mass spectrometry (Q-TOF MS).

The adulteration of dietary supplements with drugs is potentially dangerous for human health. In this study, a method was used to test simultaneously for the presence of three synthetic PDE-5 inhibitors (sildenafil, vardenafil and tadalafil), and sibutramine and its two major metabolites (N-desmethylsibutramine and N-didesmethylsibutramine) using ultra-performance liquid chromatography (UPLC) coupled with quadrupole-time-of-flight mass spectrometry (Q-TOF MS) in dietary supplements. This approach with UPLC/Q-TOF MS uses the high accurate mass of six compounds for identification and has a short run time. The recovery was from 87% to 113%; precision was less than 12.8%. The limit of detection and limit of quantification were from 0.4 to 2.0 µg kg(-1) and from 1.3 to 6.0 µg kg(-1), respectively. This method allows easy and fast analysis and detection of diverse adulterants.

Dietary combination effects of conjugated linoleic acid and flaxseed or fish oil on the concentration of linoleic and arachidonic acid in poultry meat.

This study was conducted to determine the effects of the combination of dietary conjugated linoleic acid (CLA) and n-3 fatty acids on the linoleic acid (C18:2n-6) and arachidonic acid (C20:4n-6) concentrations of broiler chicken breast and thigh muscles. One hundred and twenty broilers were raised to 6 wk of age. All chicks were fed a basal corn-soybean meal diet containing 5 different fat sources at an inclusion level of 2% total fat: 1) CLA, 2) flaxseed oil, 3) menhaden fish oil, 4) CLA and flaxseed oil, and 5) CLA and menhaden fish oil. Eight broilers from each treatment were processed at 4 and 6 wk of age. Breast and thigh muscle samples were collected and analyzed for total fat content and fatty acid composition. The results showed that broilers from the CLA and fish oil treatment had lower arachidonic acid concentrations in both breast and thigh muscles than those fed the flaxseed oil diet or the CLA and flaxseed oil diet (P < 0.05). The arachidonic acid concentration and n-6:n-3 ratio of breast and thigh samples from the menhaden fish oil diet were similar to those of the CLA and fish oil diet (P > 0.05), but the inclusion of linoleic acid into chicken thigh muscles of broilers fed the CLA and menhaden fish oil diet improved significantly when compared with that of the diet containing menhaden fish oil only. Thus, the combination of CLA and menhaden fish oil is recommended to reduce the concentrations of linoleic and arachidonic acids in broiler chicken breast and thigh muscles.

Suppression of inflammatory responses by celastrol, a quinone methide triterpenoid isolated from Celastrus regelii.

BACKGROUND: Celastrol, a quinone methide triterpenoid isolated from the Celastraceae family, exhibits various biological properties, including chemopreventive, antioxidant and neuroprotective effects. In this study, we showed that celastrol inhibits inflammatory reactions in macrophages and protects mice from skin inflammation. MATERIALS AND METHODS: Anti-inflammatory effects of celastrol (0-1 microM) were examined in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. To investigate the effects of celastrol (0-50 microg per mice) in vivo, activation of myeloperoxidase (MPO) and histological assessment were examined in the 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced mouse ear oedema model. RESULTS: Our in vitro experiments showed that celastrol suppressed not only LPS-stimulated generation of nitric oxide and prostaglandin E(2), but also expression of inducible nitric oxide synthase and cyclooxygenase-2 in RAW264.7 cells. Similarly, celastrol inhibited LPS-induced production of inflammatory cytokines, including tumour necrosis factor-alpha and interleukin-6. In an animal model, celastrol protected mice from TPA-induced ear oedema, possibly by inhibiting MPO activity and production of inflammatory cytokines. CONCLUSIONS: Our data suggest that celastrol inhibits the production of inflammatory mediators and is a potential target for the treatment of various inflammatory diseases.