Iron (Fe) is an essential metal in biological processes, which maintains a homeostasis in the human body. Divalent metal transporter 1 (DMT1) has been known as an iron transporting membrane protein, which is involved in the uptake Fe at the apical portion of intestinal epithelium, and may transport Fe across the membrane of acidified endosome in peripheral tissues. In this study, we studied the tissue distribution of DMT1 in the Fe supplemented (FeS) diet fed rats, and the regulation of DMT1 expression by depleting body Fe. Sprague-Dawley rats were divided into two groups, and fed FeS (120 mg Fe/kg) diet or Fe deficient (FeD, 2∼6 mg Fe/kg) diet for 4 weeks. The evaluation of body Fe status was monitored by measuring sFe, UIBC and tissue Fe concentration. Additionally, DMT1 mRNA levels were analyzed in the peripheral tissues by using the quantitative real time RT-PCR method. In the FeS diet fed rats, the tissue Fe was maintained at a relatively high level, and DMT1 was eventually expressed in all tissues studied. DMT1 was highly expressed in the testis, kidney and spleen, while a moderate levels of DMT1 expression was detected in the brain, liver and heart. In the digestive system, the highest level of DMT1 was found in the duodenum. Feeding the FeD diet caused a reduced body weight gain and depletion of body Fe with finding of decreased sFe, increased UIBC and decreased tissue Fe concentration. The depletion of body Fe upregulated DMT1 expression in the peripheral tissue. The expression of DMT1 was very sensitive to the body Fe depletion in the small intestine, especially in the duodenum, showing dramatically higher levels in the FeD rats than those of the FeS group. In the FeD diet fed animals, the expression of DMT1 was low significantly in other tissues compared with the duodenum. The expression of DMT1, however, was 60∼120% higher in the testis, kidney and spleen, and 30∼50% higher in the lung, liver and heart, compared to the FeS diet fed rats. In summary, DMT1 expression was ubiquitous in mammalian tissue, and the level of expression was the organ-dependent. The expression of DMT1 in peripheral tissues was upregulated by depletion of body Fe. Duodenum was the most sensitive tissue among organs studied during Fe depletion, and expressed the greatest level of DMT1, while other tissues were less higher than in duodenum. This study supports that DMT1 plays a role in maintaining the body Fe level through intestinal uptake as well as homeostasis of Fe in the peripheral tissue.
An, Hye-Suk;Han, Hee-Chang;Lee, Sun-Min;Park, Taesun;Park, Kun-Koo;Kim, Ha-Won
Proceedings of the Korean Society of Applied Pharmacology
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2001.11a
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pp.102-102
/
2001
Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.
The spermiogenesis of a Korean octopus, Octopus minor, inhabiting western of Korea Sea was observed by electron microscopy . The obtained results are as follows: The spermiogenesis of Octopus miner proceeds through four stages; early- , mid- , and late-spermatid, and mature sperm. An early spermatid is a spherical cell looking light due to the low electron density. The acrosome formed from Golgi complex of the upper nucleus looks dark due to the high electron density. The extra-nuclear rod (enr) stemming from proximal centriole is transformed from round shape into oval shape, elongating to the upper nucleus. In our observation, the axoneme was being formed from distal centriole, and the manchette composed of a number of microtubules is also found around nuclear membrane. In a mid-spermatid, chromatins in the nucleus contract shaping fine threads, and the manchette is also observed around nuclear membrane. Especially, the spherical acrosome is transformed into long oval one which is tinged with a number of horizontal stripes and has the middle electron density. In a late-spermatid, chromatins in the nucleus contract thick and short. Furthermore, the mitochondrial sleeve, in which the axoneme is surrounded with mitochondria, is observed at middle piece. The axoneme has a typical structure of 9+2 and around it, 9 coarse fibers are observed. Also in the acrosome cavity of mature sperm, horizontal striation is found. However, regularly spaced processes are peculiarly observed in there. A sperm is about 390 fm long, whose head is bent a little like a banana while the acrosome region is helical. In the middle piece of sperm, $11\sim12$ mitochondria are surrounding coarse fibers that reach the main piece of tail, while nothing but 9+2 structured axoneme is found in the end piece.
Oxygen is required for many important aerobic cellular reactions, it may undergo electrontransfer reactions, which generate highly reactive membrane-toxic intermediates (reactive oxygen species, ROS), such as hydrogen peroxide, singlet oxygen, superoxide radical, hydroxyl radical, hydroperoxyl radical, hydroxy ion. Various mechanisms are available to protect cells against damage caused by oxidative free radicals, including scavenging enzyme systems such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). This antioxidant defense system is a very complex and finely tuned system consisting of enzymes capable of detoxifying oxygen radicals as well as low molecular weight antioxidants. In addition, repair and turnover processes help to minimize subcellular damage resulting from free radical attack. $H_2O_2$,one of the major ROS, is produced at a high rate as a product of normal aerobic metabolism. The primary cellular enzymatic defense systems against $H_2O_2$ are the glutathione redox cycle and catalase. From Northern blot analysis of total RNAs from cultured cell with $H_2O_2$ treatment, various results were obtained. Expression of Cu/Zn SOD decreased when cell passage increased, but the level of the Cu/Zn SOD was scarcely expressed in 35 passage.
Kim, Jun-Hee;Kim, Mean-Hwan;Koh, Duk-su;Park, So-Jung;Kim, Soo-Jung;Nam, Joo-Hyun;Lee, Jee-Eun;Uhm, Dae-Yong;Kim, Sung-Joon
Proceedings of the Korean Biophysical Society Conference
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2003.06a
/
pp.54-54
/
2003
Prostate gland contains neuroendocrine cells (PNECs) are playing important roles in physiological and pathophysiological processes of the prostate gland. Here, we investigated the role of purinoceptors in PNECs freshly isolated from rat ventral prostate (RPNECs) that show immunoreactivity to chromogranin A. Fura-2 ratiometry revealed that ATP evokes both fast Ca$\^$2+/ influx and store Ca$\^$2+/ release in RPNECs. A whole-cell patch clamp study demonstrated fast inactivating cationic current activated by ATP or by ${\alpha}$,${\beta}$-MeATP, which was blocked by ATP-TNP. The activation of P2X inward current was tightly associated with a sharp increase in [Ca$\^$2+/]$\sub$c/. The presence of P2X1/3 subtypes were proved by RT-PCR analysis. For the stored Ca$\^$2+/ release, ATP and UTP showed similar effects, suggesting the dominant role or P2Y2 subtypes, also confirmed by RT-PCR. Both P2X (${\alpha}$,${\beta}$-MeATP) and P2Y (UTP) stimulation induced changes in the cell morphology (initial shrinkage and blob formation on the surface) reversibly. Exocytotic membrane trafficking events were monitored with the membrane-bound fluorescent dye, FM1-43 using confocal microscopy. In spite of the similar Ca$\^$2+/ responses, UTP was far less effective in triggering exocytosis than ${\alpha}$,${\beta}$ -MeATP. Since serotonin is reportedly stored in the secretory granule of PNECs, we directly examined whether the aforementioned agonists elicit release of serotonin using carbon fiber electrode-amperometry. In accordance with the results of FM1 -43 experiments, ${\alpha}$,${\beta}$-MeATP efficiently evoke serotonin secretion while not with UTP. In summary, the P2X-mediated Ca$\^$2+/ influx plays crucial roles in the exocytosis of RPNECs. Although a global increase in [Ca$\^$2+]$\sub$c/ might be related with the morphological changes, a sharp rise of [Ca$\^$2+/]$\sub$c/ in the putative sub-plasmalemmal ‘microdomains’ might be a decisive factor for the exocytosis.
It is of special interest in our membrane separation technology due to its low energy consumption and cost, relatively simple equipment, low investment and operation cost, et al. Full scale utilization of such processes can be widely utilized to the various fields. Using the difference of permeability of gas molecules between the filter layers, it is able to separate effectually pure gases from the mixed gases. In this paper, the membranes of PDMS, ${\gamma}-radiated$ PDMS, PTFE, PTFE-X are chosen to develop the predictive model for the separation of pure gases such as oxygen, nitrogen, hydrogen, and other gases from mixed gases. By utilizing the thermodynamic gas properties($\sigma$, $\varepsilon/k$) and experimental data of gas transport characteristics for different polymer membranes, it is able to develop the predictive model equation under the influence of temperature, pressure and polymer characteristics. Predictive model developed in this research showed good agreement with experimental data of gas permeability characteristics for develop four different polymer membranes. The proposed model can also be extended to the general equation for predicting the separation of gases based on the properties of polymeric membranes.
Journal of the Korean Society for Environmental Technology
/
v.19
no.6
/
pp.570-575
/
2018
The various of raw wastewater inflows to the industrial wastewater treatment plants everyday. This makes it difficult to operate the coagulation and flocculation efficiently as a pretreatment process for the industrial wastewater treatment. Additionally, it causes loads on evaporation and membrane facilities which are the posttreatment and alternative processes. For this reason, this study sampled raw wastewater before and after the NaOH injection. An experiment was conducted to compare the coagulation characteristics according to the methods of coagulant injection (single injection, simultaneous injection, consecutive injection, and inverse injection) aiming at efficient treatment of industrial wastewater. The coagulation experiment was conducted using a jar tester. The TDS removal efficiency by the coagulant injection methods increased in the order of consecutive injection (2.8 %) < single injection (3.9 %) < simultaneous injection(8.1 %) < inverse injection(9.6 %); the TOC removal efficiency grew in the order of single injection (84.3 %) < inverse injection (86.2 %) < consecutive injection (88.6 %) < simultaneous injection (89.1 %); the turbidity removal efficiency grew from single injection (99.7 %) < consecutive injection (99.8 %) ${\fallingdotseq}$ inverse injection (99.8 %) < simultaneous injection (99.9 %). These results show that the simultaneous inorganic coagulant-polymer coagulant injection method was more efficient than the consecutive injection method (inorganic coagulant ${\rightarrow}$ polymer coagulant).
Donggeun Yoo;Sohyeong Oh;Sunggi Jung;Jihong Jeong;Kwonpil Park
Korean Chemical Engineering Research
/
v.61
no.1
/
pp.58-61
/
2023
The activation process is essential for PEMFC to improve initial performance. The most commonly used activation method is a voltage change (load change) method, which may accompany degradation of the electrode catalyst if excessively performed. In many activation processes, the voltage change range is activated in a wide range from 0.4 V to OCV, and research is needed to reduce the voltage change range in order to prevent electrode catalyst degradation and shorten the activation time. Therefore, in this study, when the activation voltage range was 0.4~0.6 V, 0.4~0.8 V, and 0.4~OCV, we tried to research and develop an effective activation method by analyzing the performance and characteristics of the electrode and polymer membrane. The performance improvement was the lowest in the activation with a wide voltage range from 0.4 V to the highest OCV, and the performance decreased by 10% when activated for 56 cycles. The 0.4~0.6 V activation cycle showed the highest performance improvement up to 20% and the smallest decrease in performance due to overactivation, indicating that it is optimal method.
Song, Dong-Keun;Kater, Stanley B.;Mykles, Donald L.
The Korean Journal of Pharmacology
/
v.29
no.2
/
pp.165-174
/
1993
$Ca^{2+}$ is an important regulator of neurite elongation and growth cone movements but the mechanism(s) mediating these $Ca^{2+}-dependent$ effects is unclear. Since cytoskeletal proteins are rapidly degraded by $Ca^{2+}-dependent $ proteinases (calpains) in vitro and in vivo, we investigated whether $Ca^{2+}-induced$ pruning or regression of neuronal processes is mediated by calpains. Isolated hippocampal pyramidal-like neurons were cultured and the ability of the membrane-permeable calpain inhibitors EST (etyl (+)-(2S,3S)-3-[(S)-methyl-1-(3-methlbutylcarbamoyl)-butylcarbamoyl]2-oxiranecarboxylate) and MDL28170 (carbobenzoxyl-Val-Phe-H) to block the $Ca^{2+}$ ionophore A23187-induced suppression in neurite outgrowth was investigated. Addition of 100 nM A23187 to the culture medium resulted in a retraction of dendrites without altering axonal elongation. The addition of 300 nM A23187 to the culture medium resulted in a signiciant decrease in the rate of axonal elongation as well as a retraction of dendritic processes. Administration of EST $(5\;or\;20{\mu}M)$ to the culture medium completely blocked the pruning effect of 100 nM A23187 on dendrites and of 300 nM A23187 on axons, while EST alone did not significantly affect neurite outgrowth rate. MDL 28170 $(20\;{\mu}M)$ showed the same effect as EST in preventing ionophore-induced pruning of dendrites and axons at 100 nM and 300 nM concentrations, respectively, of A23187. EST $(20\;{\mu}M)$ did not block the A23187-induced rise of $[Ca^{2+}]_{i}$ as measured with fura-2. These results show that $Ca^{2+}-induced$ pruning of neurites in isolated hippocampal pyramidal neurons is mediated by calpains.
In this paper, the energy efficiency of wastewater utilities was evaluated to explore ways to save energy via operational measures. The correlation of each wastewater characteristic parameter to energy was assessed to find a set of parameters that explained most of the variations in energy use among utilities. The results show that increases in inflow, influent COD concentration, and ratio of advanced treatment generally increased the energy use. On the other hand, increases in load factor (influentaverage flow/design flow) reduced the energy use. In the regression analysis, the energy efficiency was highest in the A2O advanced process. On the other hand, the membrane process (among the advanced processes) and the contacted aeration process (among the secondary processes) require more efforts in saving energy. However, the data base system related to energy use must be supplemented in order for more accurate analysis of energy consumption in wastewater treatment facilities. In particular, i) electricity consumption of relay pumps and, ii) energy usage per unit process, iii) pump power usage to discharge treated wastewater in a long distance, if necessary, and iv) alternative energy production and utilization status must be recorded. By utilizing the results of the analysis conducted in this study, it is possible to quantify a level of energy savings needed and establish customized energy saving measures to achieve a certain target level for benchmarking a successful case of wastewater utilities.
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