A promoter that is inducible by paraquat and menadione, the superoxide generators, independently of soxRS has been found in front of the sufABCDSE operon in Escherichia coli. Based on the observation that SufA is a holomog of IscA that functions in the assembly of iron sulfur cluster and the sufA promoter (sufAp) contains a putative Fur-binding consensus, we investigated whether this gene is regulated by Fur, a ferric uptake regulator, When examined in several sufAp-lacZ chromosomal fusion strains, sufAp was induced by EDTA, an iron chelator and a well-known Fur-inducer, The basal level of sufA expression increased dramatically in fur mutant, suggesting repression of sufAp by Fur. The derepression in fur mutant and EDTA-induction of sufA expression required nucleotides up to -61, where a putative Fur box is located. Purified Fur protein bound to the DNA fragment containing the putative Fur box between -35 and -10 promoter elements. The regulation by Fur and menadione induction of sufAp acted independently. The rpoS mutation increased sufA induction by menadione, suggesting that the stationary sigma factor RpoS acts negatively on sufA induction.
This study was taken to examine serum components concentrations and electrophoretic patterns of female tilapia(Oreochromis niloticus) living in 0$\textperthousand$, 10$\textperthousand$, 20$\textperthousand$, and 30$\textperthousand$ salt concentrations, respectively. The results obtained in these experiments were summarized as follows. The level of albumin and total protein showed changes in each salinity, but didn't significantly(P<0.05) change in Oreochromis niloticus. The level of BUN didn't significantly(P<0.05) change. When fish were adapted from 0$\textperthousand$ to 10$\textperthousand$, 20$\textperthousand$ and 30$\textperthousand$, each calcium level in every salinity groups showed less than that of control, and didn't significantly change in 10$\textperthousand$, 20$\textperthousand$, 30$\textperthousand$ salinity. The level of calcium didn't significantly(P<0.05) change in each salinity. In 20$\textperthousand$ salinity, the level of cholesterol was at the highest peak. When fish were adapted from 0$\textperthousand$ to 10$\textperthousand$, 20$\textperthousand$ and 30$\textperthousand$, each glucose level gradually decreased. When fish were adapted from 0$\textperthousand$ to 10$\textperthousand$, 20$\textperthousand$ and 30$\textperthousand$, each glucose level gradually decreased. When fish were adapted from 0$\textperthousand$ to 10$\textperthousand$, 20$\textperthousand$ and 30$\textperthousand$. In 30$\textperthousand$ salinity, the level of alkaline phosphatase was at the highest peak. The level of serum enzyme such as SGOT and SGPT was higher in seawater-adapted group than in freshwater group. The level of phosphorus chnage significantly(P<0.05) in each salinity. Correlation coefficient between serum albumin and glucose in 0$\textperthousand$ was +0.924. Correlation coefficient between serum SGOT and SGPT of individuals in 0$\textperthousand$ was +0.917. Fraction 1 of transferrin patterns of tilapia(Oreochromis niloticus) adapted in seawater was much thicker than that of transferrin patterns of individuals adapted in freshwater. Also fraction No. a wasn't observed in some individuals adapted in freshwater. These results showed that transferrin adapted in seawater relatively increased. Slight differences, that is, showed to be observed in total iron binding capacityand iron saturatin rate between tilapia adapted in freshwater and in seawater. The increase in total iron binding capacity was attributed to a rise in transferrin pressent in the first fraction of serum protein adapted in seawater. Accordingly, the serum iron levles seemed to be related to salinity($\textperthousand$).
Proceedings of the Korean Biophysical Society Conference
/
2003.06a
/
pp.70-70
/
2003
Active sites and substrate bindings of 1-aminoxyclopropane-1-carboxylate oxidase (MD-ACO1) catalyzing the oxidative conversion of ACC to ethylene have been determined based on site-directed mutagenesis and comparative modeling methods. Molecular modeling based on the crystal structure of Isopenicillin N synthase (IPNS) provided MD-ACO1 structure. MD-ACO1 protein folds into a compact jelly roll shape, consisting of 9 ${\alpha}$-helices, 10 ${\beta}$-strands and several long loops. The MD-ACO1/ACC/Fe(II)/Ascorbate complex conformation was determined from automated docking program, AUTODOCK. The MD-ACO1/Fell complex model was consistent with well known binding motif information (HIS177-ASP179-HIS234). The cosubstrate, ascorbate is placed between iron binding pocket and Arg244 of MD-ACO1 enzyme, supporting the critical role of Arg244 for generating reaction product. These findings are strongly supported by previous biochemical data as well as site-directed mutagenesis data. The structure of enzyme/substrate suggests the structural mechanism for the biochemical role as well as substrate specificity of MD-ACO1 enzyme.
Ovotransferrin (OTF), an egg protein known as transferrin family protein, possess strong antimicrobial and antioxidant activity. This is because OTF has two iron binding sites, so it has a strong metal chelating ability. The present study aimed to evaluate the improved immune-enhancing activities of OTF hydrolysates produced using bromelain, pancreatin, and papain. The effects of OTF hydrolysates on the production and secretion of pro-inflammatory mediators in RAW 264.7 macrophages were confirmed. The production of nitric oxide (NO) was evaluated using Griess reagent and the expression of inducible nitric oxide synthase (iNOS) were evaluated using quantitative real-time polymerase chain reaction (PCR). And the production of pro-inflammatory cytokines (tumor necrosis factor [TNF]-α and interleukin [IL]-6) and the phagocytic activity of macrophages were evaluated using an ELISA assay and neutral red uptake assay, respectively. All OTF hydrolysates enhanced NO production by increasing iNOS mRNA expression. Treating RAW 264.7 macrophages with OTF hydrolysates increased the production of pro-inflammatory cytokines and the phagocytic activity. The production of NO and pro-inflammatory cytokines induced by OTF hydrolysates was inhibited by the addition of specific mitogen-activated protein kinase (MAPK) inhibitors. In conclusion, results indicated that all OTF hydrolysates activated RAW 264.7 macrophages by activating MAPK signaling pathway.
Effects of dietary ${\delta}$-aminolevulinic acid (ALA) supplementation on serum iron status, blood characteristics, egg production and quality were examined in laying hens in an 8-week feeding trail. Two hundred and forty (Hy-line brown, 40-week-old) layers were randomly assigned to four dietary treatments with ten replications (six layers in adjacent three cages). Dietary treatments included: 1) CON (basal diet), 2) ALA1 (CON+ALA 5 ppm), 3) ALA2 (CON+ALA 10 ppm) and 4) ALA3 (CON+ALA 15 ppm). All nutrient levels of diets were formulated to meet or exceed NRC (1994) recommendations for laying hens. During the entire experimental period, differences of serum iron concentration and total iron binding capacity (TIBC) were significantly increased in ALA1 supplemented treatment (quadratic effect, p<0.05). The difference of total protein between 8 and 0 weeks was significantly higher in ALA2 treatment than CON treatment (quadratic effect, p<0.05). No significant effects were observed on hemoglobin, WBC, RBC, lymphocyte and albumin concentrations. Egg production and egg weight were not influenced by the ALA supplementation. Egg yolk index was also significantly higher in ALA3 treatment than CON treatment at the end of 4 and 8 weeks (linear effect, p<0.05). Haugh unit was increased in ALA3 treatment compared to CON and ALA1 treatments at the end of 8 weeks (linear effect, p<0.05). However, egg shell thickness, breaking strength and yolk color unit were not affected by the ALA supplementation. In conclusion, dietary ALA supplementation at a level of 5 ppm can affect iron concentration in serum while higher levels (10 or 15 ppm) have some beneficial influences on blood profiles and egg quality.
Jo, Seung-Hyun;Kwon, Suk-Yoon;Park, Doo-Sang;Yang, Kyoung-Sil;Kim, Jae-Whune;Lee, Ki-Teak;Kwak, Sang-Soo;Lee, Haeng-Soon
Biotechnology and Bioprocess Engineering:BBE
/
v.11
no.5
/
pp.442-448
/
2006
Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human, including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption. We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven by an oxidative stress-inducible SWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf) was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification.
This study was designed to investigate the effects of iron supplementation and nutrition education on the iron status and anemia of high school girls. The subjects resided in Ulsan city in Korea and were already diagnosed as having anemia or iron deficiency. Over a period of three months, one iron tablet (80 mg Fe as ferrous sulfate/day) was administered to the iron deficient subjects and two tablets (160 mg Fe as ferrous sulfate/day) were administered to the anemia subjects. The average height and weight of anemia subjects were 161.24 $\pm$ 4.50 cm and 50.87 $\pm$ 5.86 kg, respectively. The average BMI (kg/$m^2$ )was 19.58 $\pm$ 2.03 and the PIBW(percent ideal body weight) were 92.52 $\pm$ 9.84%. Except for vitamin A and vitamin C intakes, the intake levels of all other nutrients were below the RDA. Total calorie intakes of anemia subjects were 73.5% of RDA. The iron intakes of subjects from food were 69. 1% of RDA and the Ca intakes were 59.1% of RDA. The basal hemoglobin(Hb) concentration of anemia subjects averaged 10.77 $\pm$ 1.33 g/dl, and this increased significantly (p < 0.001) to 12.12 $\pm$ 1.08 g/dl, after iron supplementation. The basal ferritin, and transferrin saturations {TS (%)}of anemia subjects were 12.51 $\pm$ 15.19 ng/$m\ell$ and 8.43 $\pm$ 7.56%, respectively, and these significantly increased to 20.59 $\pm$ 22.39 ng/$m\ell$ and 15.56 $\pm$ 12.87%, respectively. The level of total iron binding protein (TIBC) significantly decreased from the initial 486.80 $\pm$ 70.16 $\mu\textrm{g}$/dl to 417.86 $\pm$ 67.73 $\mu\textrm{g}$/dl (p < 0.001) after iron supplementation. For the iron deficiency subjects, the ferritin, iron and TS(%) levels were increased significantly (p < 0.001) and the TIBC levels were significantly (p <0.001) decreased after iron supplementation. Anemia symptoms such as 'Feeling blue (p<0.05)', 'Decreased ability to concentrate (p<0.001)' and 'Poor memory (p<0.05)' improved significantly after iron supplementation in the anemia subjects. The number of tablets administered was positively correlated with changes in serum hemoglobin (t=0.194, p< 0.01), serum ferritin (t=0.181, p<0.01), TS(%) (t=0.141, p<0.05), and hematocrit (t=0.254, p<0.01), and was negatively correlated with changes in TIBC (t=-0.143. p<0.05) and red cell distribution width (RDW, t=-0.140, p<0.05). In conclusion, daily iron supplementation was effective in improving the iron status and reducing symptoms of anemia in high school girls. (Korean J Nutrition 35 (9) : 943~951,2002)
Solute carrier 40A1 (SLC40A1) encodes ferroportin, which is the only known transmembrane protein that exports elemental iron from mammalian cells and is essential for iron homeostasis. Mutations in SLC40A1 are associated with iron-overload disorders. In addition to ferroportin diseases, SLC40A1 expression is downregulated in various cancer types. Despite the clinical significance of the SLC40A1 transporter, only a few studies have investigated genetic variants in SLC40A1. The present study was performed to identify genetic variations in the SLC40A1 promoter and functionally characterize each variant using in vitro assays. We investigated four haplotypes and five variants in the SLC40A1 promoter. We observed that haplotype 3 (H3) had significantly lower promoter activity than H1, whereas the activity of H4 was significantly higher than that of H1. Luciferase activity of H2 was comparable to that of H1. In addition, four variants of SLC40A1, c.-1355G>C, c.-662C>T, c.-98G>C, and c.-8C>G, showed significantly increased luciferase activity compared to the wild type (WT), whereas c.-750G>A showed significantly decreased luciferase activity compared to the WT. Three transcription factors, cAMP response element-binding protein-1 (CREB-1), chicken ovalbumin upstream promoter transcription factor 1, and hepatic leukemia factor (HLF), were predicted to bind to the promoter regions of SLC40A1 near c.-662C>T, c.-98G>C, and c.-8C>G, respectively. Among these, CREB1 and HLF bound more strongly to the variant sequences than to the WT and functioned as activators of SLC40A1 transcription. Collectively, our findings indicate that the two SLC40A1 promoter haplotypes affect SLC40A1 transcription, which is regulated by CREB-1 and HLF.
This study is to investigate the status of anemia, especially iron deficiency anemia among pre-school children in rural area in Korea. The survey was conducted in Sang-dae Ri, Yusong Myon, Daedok Gun, Chung Chong Nam-Do from July 30 th to August 12th, 1968. The measurements were done of height, weight, hematologist and biochemical levels on ninety-two pre-school children, 47 male, and 45 female, one to six years of age. Hemoglobin was determined by the method of cyanmethemoglobin and hematocrit by micro hematocrit centrifuge. The determination of serum iron, iron-binding capacity was done by the method of Ramsay using bathophenanthroline and the serum albumin was determined by Biuret Reaction. The results of this study are as follows: 1) 54.4 percent of the pre-school children weighed less than 90 percent of the Korean General Standard Weight level. 2) The average hemoglobin level was $11.0{\pm}1.57gm/100ml$, 38.0 percent of the children were anemic with less than 1.0gm/100ml. Of the anemic children 60 percent were below the Korean General Standard Weight level. 3) 27.5 percent of the pre-school children were found to have below 32 percent of a hematocrit values and 28.0 percent showed less than 33 percent in M.C.H.C. These results showed that the incidence of hypochromic anemia in these pre-school children was high. 4) 37.9 percent of these children had a serum iron level less than $50{\mu}g/100ml\;and\;31.0\;percent\;had\;a\;TIBC\;above\;400{\mu}g$ while 48.3 percent showed a transferrin saturation lower than 15 percent. On the basis of these findings, it is concluded than the cause of this anemia was iron deficiency. 5) In this group there was a little evidence of low total serum protein levels. However, 10.4 percent of the children had a deficient serum albumin level, below 2.80 gm/100ml while 51.7 percent had a low level, less than 3.50gm/100ml, and 34.5 percent of the children had a low level of TIBC, less than $350{\mu}g/100ml$, and considering these facts, it is suggested that some of the anemias have a multiple causes through protein deficiency and repeated chronic infection apart from iron deficiency.
Choi, Joon Young;Kim, Sang Eun;Lee, Kyung Han;Kim, Byung-Tae
The Korean Journal of Nuclear Medicine
/
v.32
no.1
/
pp.114-119
/
1998
We present a case of a young female patient with fulminant hepatitis who showed an altered biodistribution of Ga-67, after being scanned twice at 10 month intervals. On initial scan, uplake of Ga-67 was increased in the liver, kidneys, and skeletons. Increased hepatic Ga-67 uptake may be explained by increased transferrin unbound Ga-67 that was taken up by the inflamed liver. The saturation of iron-binding proteins due to multiple transfusions may lead to increased renal and skeletal Ga-67 uptake. On follow-up scan hepatic Ga-67 uptake was markedly increased. Also increased Ga-67 uptake in the axial skeleton and normalized renal uptake were shown. The findings were consistent with iron deficiency anemia. This case demonstrates altered Ga-67 biodistribution associated with multiple transfusions, fulminant hepatitis, and iron deficiency anemia.
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