These studies were done to find out any difference, ultrastructural, physical or chemical, between the shells of diapausing and non-diapausing eggs of the silkworm, Bombyx mori L. 1. From the electron-microscopic observation, the egg shells have four distinctive layers. In addition to the four layers, the shells in the diapausing eggs has another layer with low electron density on its surface. 2. The permeability of the egg shell to hydrochloride was much lower in diapausing egg than in non-diapausing egg. Also the permeability changed in the opposite directions with the egg age: the diapausing eggs decreased while non-diapausing ones increased. 3. The permeability increased when the diapausing egg shell was treated with HCl. When they were treated with ether, however, the increase in permeability was much smaller. It seems there was an ether soluble material involved in the content of the egg shell. 4. The diapausing eggs were also much more resistant to desiccation than the non-diapausing ones. The former, when treated with HCl or chilling, became less resistant to desiccation. 5. The positive histochemical response of the egg shell to PAS-Alcian blue and protein stainings suggests presence of abundant proteins and carbohydrates in the egg shell. On the other hand, the staining response to lipid was more positive in the inner layers than in the outer layer of the shell. 6. The egg shell adhesives seems to be mucopolysaccharides produced by colleterial glands, since the oviposited eggs showed a positive responses to carbohydrate and negative to lipid-staining chemicals, but not the mature oocytes in the ovarioles. 7. There were two bands on the electrophoretic pattern of the SH proteins extracted from the egg shells both in the diapausing egg and non-diapausing one: a slow moving major component and a fast moving minor one. However, the electrophoretic mobility showed a difference in the minor components between them. It is evident that the fast moving minor one of non-diapausing egg ran a little further than that of diapausing egg. 8. In amino acids analysis, no significant differences were found in their composition between diapausing and non-diapausing egg and SH proteins contain relatively more glycine and less cystine.
Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.
Purpose: Concerns about accelerated aging are regularly being discussed. This study analyzed the dietary status and nutritional intake of college students who are about to enter their 30s and 40s. We further suggest ways to eat healthily. Methods: Totally, 67 students attending Daegu Catholic University were provided with a three-day meal record and analyzed. Results: The average demographics of the subjects were as follows: age 23.2 ± 2.0 years, height 165.1 ± 9.0 cm, weight 65.4 ± 13.6 kg, and BMI 23.5 ± 3.9 kg/m2. Overall, 76.3% of the subjects skipped breakfast. The food intake evaluation according to the food group intake pattern and the recommended number of servings determined that 38.3% of the subjects did not consume fruits and dairy products (GMVFDS = 111001), and both men and women lacked all food groups. Analysis of the nutrient intake state revealed lesser nutrient intake than the estimated energy need. Evaluation of the %RDA found that consumption of phosphorus was above the recommended amount, whereas all other nutrients were below the recommended amount. Men had a higher intake rate of phosphorus (p < 0.01), iron (p < 0.001), and sodium (p < 0.05) than women, whereas the intake rate of dietary fiber was higher in women (p < 0.05). Evaluation of the food intake by 22 food categories revealed that intake of regular grains was more in men than in women (p < 0.05) whereas women consumed significantly more mixed grains (p< 0.01). In protein foods, men consumed more meat (p < 0.01), while women consumed more eggs (p < 0.05) and beans (p < 0.05). Evaluating the fruit intake, juices were more frequently consumed by men than by women (p < 0.05). No differences were obtained in food intake and nutrient intake status according to obesity. Conclusion: Based on these results, there is an urgent requirement for attention and support for university cafeterias in order to induce changes in the eating habits of college students. Among other initiatives, this can be achieved by providing diet improvement programs and menus that consider food preferences.
The physiological and biochemical role of potassium for upland crops according to recent research reports and the nutritional status of potassium in Korea were reviewed. Since physical and chemical characteristics of potassium ion are different from those of sodium, potassium can not completely be replaced by sodium and replacement must be limited to minimum possible functional area. Specific roles of potassium seem to keep fine structure of biological membranes such as thylacoid membrane of chloroplast in the most efficient form and to be allosteric effector and conformation controller of various enzymes principally in carbohydrate and protein metabolism. Potassium is essential to improve the efficiency of phoro- and oxidative- phosphorylation and involve deeply in all energy required metabolisms especially synthesis of organic matter and their translocation. Potassium has many important, physiological functions such as maintenance of osmotic pressure and optimum hydration of cell colloids, consequently uptake and translocation of water resulting in higher water use efficiency and of better subcellular environment for various physiological and biochemical activities. Potassium affects uptake and translocation of mineral nutrients and quality of products. potassium itself in products may become a quality criteria due to potassium essentiality for human beings. Potassium uptake is greatly decreased by low temperature and controlled by unknown feed back mechanism of potassium in plants. Thus the luxury absorption should be reconsidered. Total potassium content of upland soil in Korea is about 3% but the exchangeable one is about 0.3 me/100g soil. All upland crops require much potassium probably due to freezing and cold weather and also due to wet damage and drought caused by uneven rainfall pattern. In barley, potassium should be high at just before freezing and just after thawing and move into grain from heading for higher yield. Use efficiency of potassium was 27% for barley and 58% in old uplands, 46% in newly opened hilly lands for soybean. Soybean plant showed potassium deficiency symptom in various fields especially in newly opened hilly lands. Potassium criteria for normal growth appear 2% $K_2O$ and 1.0 K/(Ca+Mg) (content ratio) at flower bud initiation stage for soybean. Potassium requirement in plant was high in carrot, egg plant, chinese cabbage, red pepper, raddish and tomato. Potassium content in leaves was significantly correlated with yield in chinese cabbage. Sweet potato. greatly absorbed potassium subsequently affected potassium nutrition of the following crop. In the case of potassium deficiency, root showed the greatest difference in potassium content from that of normal indicating that deficiency damages root first. Potatoes and corn showed much higher potassium content in comparison with calcium and magnesium. Forage crops from ranges showed relatively high potassium content which was significantly and positively correlated with nitrogen, phosphorus and calcium content. Percentage of orchards (apple, pear, peach, grape, and orange) insufficient in potassium ranged from 16 to 25. The leaves and soils from the good apple and pear orchards showed higher potassium content than those from the poor ones. Critical ratio of $K_2O/(CaO+MgO)$ in mulberry leaves to escape from winter death of branch tip was 0.95. In the multiple croping system, exchangeable potassium in soils after one crop was affected by the previous crops and potassium uptake seemed to be related with soil organic matter providing soil moisture and aeration. Thus, the long term and quantitative investigation of various forms of potassium including total one are needed in relation to soil, weather and croping system. Potassium uptake and efficiency may be increased by topdressing, deep placement, slow-releasing or granular fertilizer application with the consideration of rainfall pattern. In all researches for nutritional explanation including potassium of crop yield reasonable and practicable nutritional indices will most easily be obtained through multifactor analysis.
A nutritional demand of oyster, Crassostrea gigas larva as part of research for improving of utilization of microalgae being used for the artificial oyster seed production. The change of body growth and biochemical compositions of larvae were investigated during larvae rearing in hatchery. The larvae were cultured in 60 M/T tank and fed mixture 6 different phytoplankton species, Isochrysis galbana (30%), Cheatoceros gracilis (20%), Pavlova lutheri (20%), Phaeodactylum triconutum (10%), Nannochryis oculata (10%) and Tetraselmis tetrathele (10%). The initial feeding amount was $0.3{\times}10^4cells/mL$ at three times a day to D-shaped larva and the feeding amount had been increased 30% gradually every two day since the larvae were raising. The larvae were developed from D shape to pediveliger stage for 12 days. The daily growth of shell length and hight were $5.8{\sim}30.8\;{\mu}m$ and $8.7{\sim}31.4\;{\mu}m$, respectively and weight gains were changed from D shape to pediveliger as follow: wet weight was $0.52{\sim}15.0\;{\mu}g/larva$, dry weight was $0.2{\sim}6.5\;{\mu}g/larva$, and ash free dry weight was $0.1{\sim}8.5\;{\mu}g/larva$. The larvae growth pattern shown a logarithmic phase from D shape to umbone stage but after that stage shown a exponential growth aspect. The crude protein, crude lipid and nitrogen free extract (NFE) of larvae during rearing periods were analyzed as $6.1{\sim}10.6%$, $0.6{\sim}1.1%$ and 1.0-2.7%, respectively. And the total amino acid contents of the larvae during rearing periods were in order as glutamic acid $1.26{\sim}2.24%$, aspartic acid $0.97{\sim}1.70%$, and methionine $0.12{\sim}0.33%$. Of the total fatty acid in the analyzed larvae, the saturated fatty acid (SSAFA) was decreased from 54.3% (D shaped larvae) to 17.1 % (pediveliger) as larvae development but the total mono-unsaturated fatty acid (${\Sigma}MOFA$) and Poly-unsaturated fatty acid (${\Sigma}PUFA$) were increased from 29.9% and 7.8% to 40.6% and 45.6%, respectively. By the way the each fatty acid of the larvae were composed of palmitic acid $9.89{\sim}36.95%$, oleic acid $12.17{\sim}32.29%$, linoleic acid $1.96{\sim}33.55%$, EPA $2.17{\sim}11.58%$ and DHA $1.95{\sim}4.51%$. As a result of this study, the larvae of oyster were demanded a various nutrients for healthy growth and the feeding control, expecially after umbone stage larvae are a rapidly growing time, is very important for success of artificial seed production.
This study was conducted to identify the correlation of bacterial phytase ($Transphos^{(R)}$) to the calcium level in feed. Of all 21-week-old 720 HyLine brown laying hens, 2 birds of similar weight were placed on each individual cage. The experiment was conducted by $3{\times}2{\times}3$ factorial design with including 3 different levels of phytase (0, 300, and 1,000 DPU/kg), 2 different levels of calcium (3.5% and 4.0%), and 3 different levels of no NPP addition 0% (0.095 NPP), 0.5% (0.185% NPP), and 1.0% (0.275% NPP). The feeding trial maintained the ME level of 2,800 kcal/kg and 16% for crude protein. The diet was fed ad libitum and 17 hours of lighting was provided throughout the experimental period. Egg production seemed to increase, in the 300 DPU of bacterial phytase added group and the cracked egg tended to reduce in Transphos added group. The egg productivity between treatment groups did not show significant difference by dietary calcium level, whereas non NPP added group (0.095% NPP) was found to be low compared to NPP added groups (P<0.05). The highest mean egg weight and the highest daily egg mass were detected in 300 DPU phytase added group. Although the mean egg weight was significantly higher in treatment groups fed with 3.5% calcium containing feeds (P<0.05), daily egg mass was no among treatment groups. The mean egg weight and daily egg mass were the lowest in non NPP added group (0.095% NPP) compared to other treatment groups (P<0.05). The feed intake showed similar pattern regardless of the bacterial phytase and calcium levels in the diet. However, the treatment groups fed diets containing NPP level of 0.275% and 0.165% showed significantly higher feed intake than the group fed with 0.095% NPP (P<0.05). Although the feed conversion was not affected by calcium and NPP levels in the diet, the most improved result was obtained from 300 DPU phytase added group (P<0.05). The eggshell breaking strength and thickness increased as dietary calcium level increase the level of calcium increases in diet. The treatment groups fed diet containing 0.275% and 0.165% NPP revealed to show improvement in eggshell breaking strength and yolk color index compared to the NPP non added (0.095% NPP) treatment group. The result of the present study suggests that the appropriate level of microbial phytase is 300 DPU and at this level, tricalciumphosphate supplementation in feed can be reduced to 40% of NRC recommendation. Higher calcium level in feed fail to show synergistic effect by adding microbial phytase.
Kim, K.S.;Chee, K.M.;Lee, S.J.;Cho, S.K.;Kim, S.S.;Lee, W.
Korean Journal of Poultry Science
/
v.18
no.2
/
pp.97-119
/
1991
Effect of Streptococcus faecium(SF) and an antibiotic, Colistin(Col), supplemented to diets singly or in combination, on the performances and changes of intestinal population of microflora of broiler chicks studied. A total of 252, day-old chicks(Arbor Acre) of mixed sex(M:F=1:1) were alloted into six groups. A diet with no Col and SF was referred as a control diet. The basal diets were added with two levels of SF, 0.04 and 0.08%, singly or in combination with Col 10ppm Another diet was prepared by adding only Col 10 ppm. Numbers of the microorganism in diets added with SF 0.04% and 0.08% were 7$\times$10$^{4}$ and 1.4$\times$10$^{5}$ /g diet respectively The diets consisting of corn and soybean meal as major ingredients were fed for a period of seven weeks . During the feeding trial, fresh excreta were sampled at the end of every week in a sterilized condition to count microbial changes from each dietary group. Microbial changes of large intestine were also measured from nine birds sacrificed at the end of the 4th and 7th weeks each time per dietary group. Excreta from all the groups were also collected quantitatively at the end of 3rd and 6th weeks to measure digestibility of the diets, At the end of 7th week, nine birds from each group were also sacrificed to measure weight changes of gastrointestinal tracts . Average body weight gains of broilers fed the diets added with SF 0.08% (2.37kg) or SF 0. 08%+col 10ppm(2.34kg) were significantly larger than that of the control(2.18kg). The weight gains of the other groups were not statistically different from that of the control Feed/gain ratios of the supplemental groups were better than that of control (P<0.05) except that of birds fed the diet added only with SF 0.04%. Digestibilities of nutrients such as dry matter, crude protein, crude fat and total carbohydrates were not altered by the consumption of the diets added with SF and/or Col throughout the whole feeding period. As expected, the numbers of Streptococci in the excreta from birds fed diets added with SF increased significantly with a statistical difference between groups with SF 0.04% and SF 0.08% most of the time. However. addition of Colistin to the diets supplemented with SF did not give any effects on the number of the microorganism. Numbers of coliforms in the excreta were apparently reduced by feeding the diets added with SF and/or Col(P<0.05). There were, however, no additive effects observed between the two feed additives in this regard when supplementing Col to the SF diets. Distributions of intestinal microflora exhibited exactly the same pattern as those of the excreta. Length of small intestine of the birds fed diets added with SF 0.08% with or without Col 10 ppm became significantly longer with a range of about 10% than those of the birds fed diets without SF. However, the empty weight of the small inestine of the former group was lighter than that of control These changes resulted in a significant reduction in weight/unit length of the intestine of the birds fed diets supplemented with Col and SF singly or in combination. In overall conclusion, diet added with SF 0.08% appeared most effective in improving broiler performances. Colistin added at a level of 10ppm was not beneficial at all in itself or in combination with SF in terms of broiler performances or changes of intestinal microflora population. The efficacy of SF and Col could be attributed to the changes of wall thickness of the small intestine.
Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.
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