Lactoferrin is an 80-kDa iron-binding glycoprotein that is found in high concentrations in human milk. Human lactoferrin (hLF) has several beneficial biological activities including immune system modulation and antimicrobial activity. In the present study, we devolved a method of hLF expression through introducing the hLF gene construct into Oriza sativa cv. Nakdong using the Agrobacterium-mediated transformation system. The expression of the hLF gene under the control of the rice glutelin promoter was detected in the seeds of transgenic rice plants. Transformed rice plants were selected on media containing herbicide(DL-phosphinothricin) and the integration of hLF cDNA was confirmed by Southern blot analysis. The expression of the full length hLF protein from the grains of transgenic rice plants was verified by Western blot analysis. The lactoferrin expression levels in the transformed rice grains determined by enzyme-linked immunosorbant assay accounted for approximately 1.5% of total soluble protein. Taken together, these data indicate that rice grains expressing hLF can be directly incorporated into infant formula and baby food.
The effect of meal service for home-staying, urban elderly with low income on their mineral status and prevalence of clinical symptoms was evaluated. One hundred. One hundred and eighty three subjects were assigned to meal-served(served) and non-served(non-served)groups. A meal containing approximately one half of the RDA for energy, protein, calcium and iron was served as lunch every day to served group at a welfare center. Dietary, biochemical and clinical data were collected before and after 6 months of meal service and the changes of parameters were analyzed with paired t-test. served female showed significantly increased intake of calcium. The mean hemoglobin, serum iron, TIBC and serum copper of female were significantly increase with meal service. The proportion of anemic female compared to reference data on hemoglobin, hematocrit and serum iron were 45.5$\%$, 29.1$\%$ and 16.4$\%$ respectively, but were lowered to 18.2$\%$, 7.3$\%$, and 5.5$\%$ after 6 months of meal service. served male showed significant increase in MCHC, serum copper, and urinary Na. The urinary Ca/Creatinine, which is often used as an index of bone resorption, was decreased significantly in served group. The decrease in the proportion of served women suffering from edema of ankle and diarrhea was most apparent and less women reported feeling clinical symptoms of dizziness, constipation, difficulty in hearing, and coughing after meal service. The symptoms of benumbness of hands and feet and coughing was lowered most among men after meal service. Clinical symptoms of non-served elderly did not show improvement in feeling clinical symptoms except slightly decreased frequency in coughing of female and buzzing sound in the ears and diarrhea of male.
A superoxide dismutase was purified 62-fold in seven steps to homogeneity from Methylobacillus sp. strain SK1, an obligate methanol-oxidizing bacterium, with a yield of 9.6%. The final specific activity was 4,831 units per milligram protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. The molecular weight of the native enzyme was estimated to be 44,000. Sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 23,100. The isoelectric point of the purified enzyme was found to be 4.4. Maximum activity of the enzyme was measured at pH 8. The enzyme was stable at pH range from 6 to 8 and at high temperature. The enzyme showed an absorption peak at 280 nm with a shoulder at 292 nm. Hydrogen peroxide and sodium azide, but not sodium cyanide, was found to inhibit the purified enzyme. The enzyme activity in cell-free extracts prepared from cells grown in manganese-rich medium, however, was not inhibited by hydrogen peroxide but inhibited by sodium azide. The activity in cell extracts from cells grown in iron-rich medium was found to be highly sensitive to hydrogen peroxide and sodium azide. One mol of native enzyme was found to contain 1.1 g-atom of iron and 0.7 g-atom of manganese. The N-terminal amino acid sequence of the purified enzyme was Ala-Tyr-Thr-Leu-Pro-Pro-Leu-Asn-Tyr-Ala-Tyr. The superoxide dismutase of Methylobacillus sp. strain SK1 was found to have antigenic sites identical to those of Methylobacillus glycogenes enzyme. The enzyme, however, shared no antigenic sites with Mycobacterium sp. strain JC1, Methylovorus sp. strain SS1, Methylobacterium sp. strain SY1, and Methylosinus trichosproium enzymes.
Background: New alternative types of pet foods such as raw and cooked homemadestyle diets containing human food ingredients have been introduced due to a trend of pet humanization and diversification of consumer needs. Objectives: To evaluate nutritional adequacy of new alternative types of dog foods containing human food ingredients as maintenance diets for dogs. Methods: Eleven homemade-style foods for adult dogs were purchased from online channel in Korea and analyzed to evaluate nutritional adequacy for adult dogs. Nutrients analyzed included crude protein, amino acids, crude fat, fatty acids, and minerals. Results: Crude protein and amino acids in all products satisfied Association of American Feed Control Officials (AAFCO) requirements. Crude fat in one of 11 products did not meet AAFCO requirements. The most deficient minerals were selenium (10 of 11, 90.9%), copper (five of 11, 45.5%), zinc (five of 11, 45.5%), potassium (three of 11, 27.3%), calcium (three of 11, 27.3%), iron (two of 11, 18.2%), and magnesium (one of 11, 9.1%). Six products were not in the range of the recommended Ca:P ratio in AAFCO dog food maintenance nutrient profiles. Conclusions: This study performed nutritional evaluation of raw and cooked homemadestyle foods as maintenance diets for adult dogs. Some nutritional inadequacies were observed including some minerals, Ca:P ratio, and omega-6:omega-3 fatty acid ratio, although three products (26.2%) satisfied the AAFCO standard except selenium. Overall, the data suggest a need for accurate nutritional adequacy statement for consumers based on proper methods to validate the formula.
Objective: This study investigated the effects of feeding anthocyanin-rich black cane treated with ferrous sulfate and molasses on animal performance, rumen fermentation, microbial composition, blood biochemical indices, and carcass characteristics in meat goats. Methods: Thirty-two Thai-native×Anglo-Nubian crossbred male goats (14.47±2.3 kg) were divided equally into two groups (n = 16) to investigate the effect of feeding diet containing 50% untreated anthocyanin-rich black cane silage (BS) vs diet containing anthocyaninrich black cane silage treated with 0.03% ferrous sulfate and 4% molasses (TBS) on average daily gain (ADG) and dry matter intake (DMI). At the end of 90 d feeding trial, the goats were slaughtered to determine blood biochemical indices, rumen fermentation, microbial composition, and carcass characteristics differences between the two dietary groups. Results: Goats fed the TBS diet had greater ADG and ADG to DMI ratio (p<0.05). TBS diet did not affect rumen fluid pH; however, goats in the TBS group had lower rumen ammonia N levels (p<0.05) and higher total volatile fatty acid concentrations (p<0.05). Goats in the TBS group had a higher (p<0.05) concentration of Ruminococcus albus but a lower (p<0.05) concentration of methanogenic bacteria. The TBS diet also resulted in lower (p<0.05) thiobarbituric acid-reactive substances concentration but higher (p<0.05) total antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase concentrations in blood plasma, while having no effect on plasma protein, glucose, lipid, immunoglobin G, alanine transaminase, and aspartate aminotransferase. Meat from goats fed the TBS diet contained more intramuscular fat (p<0.05) and was more tender (p<0.05). Conclusion: In comparison to goats fed a diet containing 50% untreated anthocyanin-rich black cane silage, feeding a diet containing 50% anthocyanin-rich black cane silage treated with 0.03% ferrous sulfate and 4% molasses improved rumen fermentation and reduced oxidative stress, resulting in higher growth and more tender meat.
The function of the [4Fe-4S] cluster containing iron (Fe-) protein in nitrogenase catalysis is to serve as the nucleotide-dependent electron donor to the MoFe protein which contains the sites for substrate binding and reduction. The ability of the Fe protein to function in this manner is dependent on its ability to adopt the appropriate conformation for productive interaction with the MoFe protein and on its ability to change redox potentials to provide the driving force required for electron transfer. The MgADP-bound (or off) conformational state of the nitrogenase Fe protein structure described reveals mechanisms for long-range communication from the nucleotide-binding sites to control affinity of association with the MoFe protein component. Two pathways, termed switches I and II, appear to be integral to this nucleotide signal transduction mechanism. In addition, the structure of the MgADP bound Fe protein provides the basis for the changes in the biophysical properties of the [4Fe-4S] observed when Fe protein binds nucleotides. The structures of the nitrogenase Fe protein with defined amino acid substitutions in the nucleotide dependent signal transduction pathways of the Switch I and Switch II have been determined by X-ray diffraction methods. These two pathways have been also implicated by site directed mutagenesis studies, structural analysis and analogies to other proteins that utilize similar nucleotide dependent signal transduction pathways. We have examined the validity of the assignment of these pathways in linking the signals generated by MgATP binding and hydrolysis to macromolecular complex formation and intermolecular electron transfer. The results provide a structural basis for the observed biophysical and biochemical properties of the Fe protein variants and interactions within the nitrogenase Fe protein-MoFe protein complex.
In order to find the feasibility of feeding raw or processed cotton (Gossypium) seed meal (CSM), 30 male crossbred lambs were randomly assigned to 5 dietary treatments with 6 animals each. Lambs were fed each of the isonitrogenous and isocaloric concentrate mixtures containing 30% deoiled groundnut cake (DGNC) (reference diet), 40% raw, 45 minute cooked, 1% $Ca(OH)_2$ or iron (1 part free gossypol:0.3 parts iron) treated CSM (replacing about 50% nitrogen moiety of reference concentrate mixture). The concentrate mixture was fed to meet 80% of CP requirements (NRC, 1985) along with ad libitum maize (Zea mays) hay for 180 days. Incorporation of raw or processed CSM did not affect the palatability of diets as evidenced by higher (p<0.01) or comparable overall daily intakes of DM and protein (CP and Digestible CP) per kg $W^{0.75}$ by lambs in comparison to reference group. Intakes of DE and ME increased (p<0.01) due to inclusion $Ca(OH)_2$ treated CSM in diets, while the intakes were lower (p<0.01) on iron treated CSM based diet when compared with reference diet. The intakes of DM, CP, TDN, DE and ME by reference and experimental lambs were higher than the requirements stipulated by NRC (1985). The growth rate was highest (p<0.01) in lambs fed on diets with cooked CSM followed by those fed raw or $Ca(OH)_2$ treated CSM based diets. The growth of lambs fed on iron treated CSM diets was depressed (p<0.01) in comparison to that of other experimental lambs probably due to increased iron intake (889 mg/kg DM of diet) which was much higher than the toxic levels of 500 ppm (NRC, 1980). The lambs on various diets utilized DM with similar efficiency while the utilization of energy (TDN, DE and ME) was more efficient (p<0.01) when the diets contained raw or cooked CSM in comparison to that of reference diet. The lambs fed raw and $Ca(OH)_2$ treated CSM incorporated diets utilized DCP with higher (p<0.01) efficiency than the lambs fed reference diet. The feed cost per kg weight gain was lower (p<0.01) on raw, cooked and $Ca(OH)_2$ treated CSM based diets in comparison to reference diet. Cooking of CSM for 45 minutes further reduced the feed cost of weight gain.
Shin, Kyung-Ok;Ha, Seo-Yeong;Shin, Seong-Beom;Kim, Jeong-Yeon;Yang, Ming
The Korean Journal of Food And Nutrition
/
v.34
no.6
/
pp.593-603
/
2021
In this study, Korean Hallabong produced in Jeju Island and coffee were grafted to prepare coffee containing Hallabong extract and the nutritional components were analyzed. As the amount of Hallabong extract increased, the water content and total polyphenol content increased. However, the crude flour, crude protein, and total flavonoid content decreased significantly. The selenium content per 100 g was 91.28 mg in the 1% Hallabong group, and the iron content was 6.84 mg in the 3% Hallabong group. As the content of Hallabong extract in coffee increased, the L-value (brightness) and b-value (yellowness) increased, but the a-value (redness) showed a tendency to decrease. In the case of DPPH(2,2-Diphenyl-1-picrylhydrazyl) radical scavenging activity, the group containing 9% of Hallabong extract showed the highest value at 47.20 μmol/g of TEAC. In particular, the ABTS(2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonate)) and DPPH radical scavenging activity were significantly increased from coffee powder containing 6% or more of Hallabong extract(p<0.05). The caffeine content decreased as the amount of Hallabong extract added to coffee increased. Therefore, when making powder coffee with Hallabong extract added, it is recommended to set the content of Hallabong extract to 6%.
Lactoferrin is an 80-kDa iron-binding glycoprotein known to exert many biological activities, such as facilitating iron absorption and having antimicrobial and anti-inflammatory effects. Rice can be a useful target for edible food plants to introduce human lactoferrin, because it has lower allergenicity and is likely to be safer than microorganisms or transgenic animals. A cDNA fragment encoding human lactoferrin (HLF) driven by the maize polyubiquitin promoter, along with herbicide resistance gene (bar) driven by CaMV 35S promoter, was introduced into rice (Oryza sativa L. cv. Dong Jin) using the Agrobacterium -mediated transformation system. Putative transformants were initially selected on the medium containing bialaphos. The stable integration of the bar and HLF genes into transgenic rice plants was further confirmed through polymerase chain reaction (PCR) and Southern blot analyses. The expression of the full length HLF protein from various tissues such as grains and young leaves of transgenic rice was verified by Western blot analysis. Analysis of progeny also demonstrated that introduced genes were stably inherited to the next generation at the Mendelian fashion.
This study examined the effects of excess intake of calcium (Ca) and iron (Fe) supplement on bone loss, nephrocalcinosis and renal function in osteoporotic model rats. Seven-week-old female rats were first fed a Ca-deficient diet for four weeks after ovariectomy operation, and then one of nine experimental diets for additional eight weeks, containing three levels of Ca, normal (0.5%) or high (1.5%) or excess (2.5%) and three levels of Fe, normal (35ppm) or high (210ppm) or excess (350ppm). The osteoporotic model rats showed a remarkable increase in body weight, serum alkaline phosphatase (ALP) and decrease in breaking force, Ca, P, Mg contents of femur. Serum Ca concentration was not significantly affected by dietary Ca and Fe levles. Liver Ca content increased in rats fed a high-and excess-Ca diet. Kidney Ca content and microscopic Ca deposition remarkably increased in osteoporotic model rats compared to control group, and showed a tendency to decrease in rats fed a excess-Ca diet. Breaking force of femur increased with increasing dietary Ca levels, but Ca, P contents of femur and serum ALP were not significantly affected by dietary Ca and Fe levels. Serum total protein decreased in rats fed a excess-Ca diet, BUN increased in rats fed a excess-Ca diet, while serum uric acid and creatinine were not significantly affected by dietary Ca levels. Urinary creatinine, GFR increased in rats fed a high-and excess-Ca, diet, and GFR was highest in rats fed a excess-Ca/excess-Fe diet. These results suggest that excess intake of Ca may increase breaking force of femur, but not increase mineral contents of femur, and decrease kidney function. Furthermore, excess intake of Fe and Ca concurrently may aggravate kidney function leading to potential health problems in ovariectomized osteoporotic model rats.
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