Purpose: The balance between synthesis and degradation of proteins plays a critical role in the maintenance of skeletal muscle mass. Mitochondrial dysfunction has been closely associated with skeletal muscle atrophy caused by aging, cancer, and chemotherapy. Polygalacin D is a saponin derivative isolated from Platycodon grandiflorum (Jacq.) A. DC. This study aimed to investigate the effects of polygalacin D on myoblast differentiation and muscle atrophy in association with mitochondrial function in in vitro and in zebrafish models in vivo. Methods: C2C12 myoblasts were cultured in differentiation media containing different concentrations of polygalacin D, followed by the immunostaining of the myotubes with myosin heavy chain (MHC). The mRNA expression of markers related to myogenesis, muscle atrophy, and mitochondrial function was determined by real-time quantitative reverse transcription polymerase chain reaction. Wild type AB* zebrafish (Danio rerio) embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without polygalacin D, and immunostained to detect slow and fast types of muscle fibers. The Tg(Xla.Eef1a1:mitoEGFP) zebrafish expressing mitochondria-targeted green fluorescent protein was used to monitor mitochondrial morphology. Results: The exposure of C2C12 myotubes to 0.1 ng/mL of polygalacin D increased the formation of MHC-positive multinucleated myotubes (≥ 8 nuclei) compared with the control. Polygalacin D significantly increased the expression of MHC isoforms (Myh1, Myh2, Myh4, and Myh7) involved in myoblast differentiation while it decreased the expression of atrophic markers including muscle RING-finger protein-1 (MuRF1), mothers against decapentaplegic homolog (Smad)2, and Smad3. In addition, polygalacin D promoted peroxisome proliferator-activated receptor-gamma coactivator (Pgc1α) expression and reduced the level of mitochondrial fission regulators such as dynamin-1-like protein (Drp1) and mitochondrial fission 1 (Fis1). In a zebrafish model of FOLFIRI-induced muscle atrophy, polygalacin D improved not only mitochondrial dysfunction but also slow and fast muscle fiber atrophy. Conclusion: These results demonstrated that polygalacin D promotes myogenesis and alleviates chemotherapy-induced muscle atrophy by improving mitochondrial function. Thus, polygalacin D could be useful as nutrition support to prevent and ameliorate muscle wasting and weakness.
The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
/
v.14
no.4
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pp.205-212
/
2009
To suggest possible to bioremediation by benthic microalgae Nitzschia sp. isolated from the Jinhae Bay, the studies investigated the effects o flight quality and quantity on the growth of Nitzschia sp. and its growth kinetics for phosphate investigated. The Nitzschia sp. was cultured under blue (450 nm), yellow (590 nm) and red wavelength (650 nm) using light emitting diode (LED) and mixed wavelengths using a fluorescent lamp. The maximum specific growth rate showed the Nitzschia sp. under blue wavelength, although photoinhibition was observed above $100\;{\mu}mol\;m^{-2}\;s^{-1}$. Mixed wavelengths were also observed by decreasing the maximum cell density from high irradiances (>$100\;{\mu}mol$ photons $m^{-2}\;s^{-1}$). The compensation photon flux density ($I_0$) calculated from the mixed wavelengths equated to a depth of 4-10 m in Jinhae Bay, and was lower in the summer season than the depth due to suspended matter (ca. 4 m). Thus, the suitable depth for maximum growth of Nitzschia sp. might be extremely limited. In the growth kinetics for phosphate, half-saturation constant ($K_s$) was similar among different wavelengths, although the maximum growth rate was varied among different wavelengths. Because the $K_s$ was high than that of the phytoplankton, Nitzschia sp. might have adapted to the high nutrient concentrations, and have effective nutrient storage in the cell quota. Thus, Nitzschia sp. may be a useful species for bioremediation of the benthic layer in polluted inner bays by means of irradiated specific wavelength as blue.
Seminal plasma(SP) is usually removed from semen that is to be cryopreserved. However, some reports indicate that SP has beneficial effects on spermatozoa during chilling and freezing. The purpose of this study was to determine the effect of SP on sperm survival by adding SP to the extender before cooling and freezing canine spermatozoa. In replicate experiments, ejaculates obtained from four healthy dogs(1-4 years old) of various breeds were pooled, centrifuged at $300{\times}g$ for 10 min at $25^{\circ}C$, and the supernatant of seminal plasma was decanted. Spermatozoa were suspended in egg yolk-Tris(EYT) buffer. The study comprised two experiments: [Exp 1] Sperm were suspended in EYT extender containing either 0, 20, 40, 80 or 100% SP and were slowly cooled to $4^{\circ}C$ for 2h or held at $25^{\circ}C$ as controls. Sperm concentration was adjusted to $2{\times}10^8/ml$. [Exp II] Sperm samples, each of which contained $1{\times}10^8/ml$, were assigned to nine groups to be frozen. In the first four groups, sperm in EYT containing either 20, 40, 80 or 100% SP were cooled to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol and then were frozen. The final concentrations of SP were 10, 20, 40 or 50%. In the other four groups, sperm in EYT alone were first cooled slowly to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol plus 10, 20, 40 or 50% SP and then were frozen. Spermatozoa, which chilled in EYT alone and diluted to contain final concentrations of EYT+0.6M glycerol without seminal plasma, and then frozen, was regarded as control. Spermatozoa were frozen at $25^{\circ}C/min$ of cooling rate in plastic straws that were suspended above liquid nitrogen and thawed in water at $38^{\circ}C$ for 1 min. Sperm survival was assayed by determining progressive motility and integrity of plasma and acrosome membranes. Progressive motility was determined by microscopic examination at $200{\times}$ magnification. Membrane integrity was assessed by use of a double fluorescent dye, and acrosome integrity by staining sperm with Pisum sativum agglutinin. The results of the first experiment showed that adding SP did not improve motility of spermatozoa compared to those incubated without SP regardless of temperature. The results of the second experiment showed that spermatozoa suspended in EYT+0.6M glycerol containing SP exhibited the higher progressive motility before being frozen(P<0.05). However, frozen-thawed spermatozoa that had suspended in EYT+0.6M glycerol containing SP showed the similar or lower viability(P<0.05). In summary, although seminal plasma did not affect spermatozoa that were chilled in EYT without cryoprotectant(CPA), addition of seminal plasma to EYT containing CPA did significantly improved progressive motility of canine spermatozoa that were chilled.
BACKGROUND: Hydroponics is one of the methods for evaluating plant production using the inorganic nutrient solutions, which is applied under the artificial light conditions of plant factory system. However, the application of the conventional inorganic nutrients for hydroponics caused several environmental problems: waste from culture mediums and high nitrate concentration in plants. Organic nutrients are generally irrigated as a supplementary fertilizer for plant growth promotion under field or greenhouse conditions. Hydroponic culture using organic nutrients derived from the agricultural by-products such as dumped stems, leaves or immature fruits is rarely considered in plant factory system. Effect of organic or conventional inorganic nutrient solutions on the growth and nutrient absorption pattern of green and red leaf lettuces was investigated in this experiment under fluorescent lamps (FL) and mixture Light-Emitting Diodes (LEDs). METHODS AND RESULTS: Single solution of tomatoes (TJ) and kales (K) deriving from agricultural by-products including leaves or stems and its mixed solution (mixture ration 1:1) with conventional inorganic Yamazaki (Y) were supplied for hydroponics under the plant factory system. The Yamazaki solution was considered as a control. 'Jeockchima' and 'Cheongchima' lettuce seedlings (Lactuca sativa L.) were used as plant materials. The seedlings which developed 2~3 true leaves were grown under the light qualities of FL and mixed LED lights of blue plus red plus white of 1:2:1 mixture in energy ratio for 35 days. Light intensity of the light sources was controlled at 180 μmol/㎡/s on the culture bed. The single and mixture nutrient solutions of organic and/or inorganic components which controlled at 1.5 dS/m EC and 5.8 pH were regularly irrigated by the deep flow technique (DFT) system on the culture gutters. Number of unfolded leaves of the seedlings grown under the single or mixed nutrient solutions were significantly increased compared to the conventional Y treatment. Leaf extension of 'Jeockchima' under the mixture LED radiation condition was not affected by Y and YK or YTJ mixture treatments. SPAD value in 'Jeockchima' leaves exposed by FL under the YK mixture medium was approximately 45 % higher than under conventional Y treatment. Otherwise, the maximum SPAD value in the leaves of 'Cheongchima' seedlings was shown in YK treatment under the mixture LED lights. NO3-N contents in Y treatment treated with inorganic nutrient at the end of the experiment were up to 75% declined rather than increased over 60 % in the K and TJ organic treatment. CONCLUSION: Growth of the seedlings was affected by the mixture treatments of the organic and inorganic solutions, although similar or lower dry weight was recorded than in the inorganic treatment Y under the plant factory system. Treatment Y containing the highest NO3-N content among the considered nutrients influenced growth increment of the seedlings comparing to the other nutrients. However effect of the higher NO3-N content in the seedling growth was different according to the light qualities considered in the experiment as shown in leaf expansion, pigmentation or dry weight promotion under the single or mixed nutrients.
Kim, Dong-Wook;Kim, Ji-Hyuk;Kang, Geun-Ho;Kang, Hwan-Ku;Lee, Sang-Jin;Lee, Won-Jun;Kim, Sang-Ho
Journal of Animal Science and Technology
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v.50
no.6
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pp.807-818
/
2008
This study was conducted to prove the optimum feeding method of Lactobacillus in broiler chickens by investigating the intestinal viability of ingested Lactobacillus and the effect of feeding levels and frequency of Lactobacillus on growth performance in broiler chickens. In experiment 1, A total of one hundred, 5 weeks old male broiler chickens(Abor Acre) were fed Lactobacillus reuteri avibro2 expressed green fluorescent protein(GFP) at 104cfu/g diet to investigate the retention time of ingested Lactobacillus in the intestine for 1 day. The percentage of Lactobacillus expressed GFP in intestinal contents was 26% at 1 day after fed Lactobacillus expressed GFP. The percentage of Lactobacillus expressed GFP in intestinal contents was decreased in length of time. In experiment 2, A total of four hundred eighty, 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each to prove the optimum feeding level of Lactobacillus. The treatments were control(free antibiotics), Lactobacillus reuteri avibro2 5.0×10cfu/mL, 5.0×103cfu/mL, and 5.0×105cfu/mL. The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). Feed conversion ratio was not significantly difference among the groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). Protein and fat digestibility in Lactobacillus 5.0×103cfu/mL and 5.0×105cfu/mL treated groups were significantly improved(P<0.05). No significant differences were observed on the availability of dry matter and crude ash in Lactobacillus treatments compared to those of control. In experiment 3, A total of six hundred 1-d-old male broiler chicks(Abor Acre) were randomly divided into 4 groups with 4 replicates of 30 birds each and were fed Lactobacillus reuteri avibro2 at intervals of 1, 2, 3, and 5 day for five weeks. Feeding level of Lactobacillus was 5.0×103cfu/mL The final body weight and body wight gain of Lactobacillus reuteri avibro2 5.0×103cfu/mL were the highest in all groups(P<0.05). The final body weight and body weight gain were significantly increased, when Lactobacillus was fed at intervals of 1 days, or 2 days. There were no significant differences in feed intake and feed conversion ratio among the all groups. The number of intestinal lactic acid bacteria in Lactobacillus treated groups tended to be improved or significantly increased as compared to that of control(P<0.05). No significant differences were observed on the number of coliform bacteria and Salmonella of ileum and cecum. Consequently, supplemental Lactobacillus influenced positive effects on the growth performance, nutrient availability and intestinal microflora. The optimum feeding level of Lactobacillus was 5.0×103cfu/mL, and the constant feeding of Lactobacillus was effective.
Development of protein resources as food has been a big issue especially in Southeast Asia region, and intake of protein is also insufficient in Korea. To cope with this shortage of protein resources and its improvement together with increased production of high nutritive animal protein, studies were carried out on feeding of Korean native goats. In the experiments were made absorption of carbohydrate and volatile fatty acid in miniature rumen, and absorption of amino acid in rumen as in vivo were conducted as part of studies on nutritional absorption in rumen. Those nutritional for improvement of feeding and management as described above are summarized as following. 1. According to the result of test on the nutritional absorption of native goat by means of miniature rumen method, absorption ratio of VFA measured at 0.5, 1 and 2 hours after injection of nutrition showed propionic acid 70-86%, acetic acid 74-87%, and lactic acid 76-89%. In the absorption of organic substances, ethyl alcohol of 0.5% showed 29-87% and lactic acid of 0.1M showed 12-27% of absorption ratio. 2. Result of absorption measurement in rumen from L-type free amino acid injection in the content of rumen vein showed lower rate at menthionine-free group compared to whole-egg amino acid injection in the content of rumen vein showed lower rate at methioine-free group compared to whole-egg amino acid group, and high absorption ratio was observed at methionine 3 times group and urea added group. Deficiency of methionine caused no change of the content in mucous membranes. 3. Absorption of amino acid in rumen muscular layer showed equal tendency as in the mucous membrane without exerting any influence of methionine deficiency. At the methionine3-times group, content of methionine and glutamine were increased by 14.7 and 4.4 times as compared to whole-egg amino acid group, an absorption ratio of glutamine, proline and valine were increased at urea added group. 4. In general, concentration of amino acid in rumen vein plasma was lower than in rumen mucous membrane and muscular layer. Absorption ratio of amino acid is decreased due to methionine deficiency, and tripling of methionine or urea adding caused increment of amino acid. Absorption pattern is thus varied depending on the composition of amino acid. 5. At the urea added group, content of ammonia-N, amino-N and urea were increased in rumen muscular layer. As the inside of goat's rumen was unable to clean thoroughly, investigation was made on remaining bacteria, however, variation of ammonia-N was affected by these bacterial content. 6. Variation in rumen structure by differential absorption of amino acid was observed by general microscope and fluorescent microscope. According to the result of observation in the methionine 3 times group, single cylinder epithelium of mucous membrane showed rather thin, and it was thick at urea added group though no significant differences existed among test groups in submucous membrane and muscular layer.
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