• BMB Reports
• /
• 제34권4호
• /
• pp.316-321
• /
• 2001

Dehydroascorbate (DHA) reductase (DHAR, EC 1.8.5.1) catalyzes the reduction of DHA to reduced ascorbate (AsA) using glutathione (GSH) as the electron donor in order to maintain an appropriate level of ascorbate in plant cells. To analyze the physiological role of DHAR in environmental stress adaptation, we developed transgenic tobacco (Nicotiana tabacum cv. Xanthi) plants that express a human DHAR gene isolated from the human fetal liver cDNA library in the chloroplasts. We also investigated the DHAR activity, levels of ascorbate, and GSH. Two transgenic plants were successfully developed by Agrobacterium-mediated transformation and were confirmed by PCR and Southern blot analysis. DHAR activity and AsA content in mature leaves of transgenic plants were approximately 1.41 and 1.95 times higher than in the non-transgenic (NT) plants, respectively In addition, the content of oxidized glutathione (GSSG) in transgenic plants was approximately 2.95 times higher than in the NT plants. The ratios of AsA to DHA and GSSG to GSH were changed by overexpression of DHAR, as expected, even though the total content of ascorbate and glutathione was not significantly changed. When tobacco leaf discs were subjected to methyl viologen at $5\;{\mu}M$, $T_0$ transgenic plants showed about a 50% reduction in membrane damage compared to the NT plants.

• Applied Microscopy
• /
• 제27권3호
• /
• pp.321-332
• /
• 1997

The dermal papilla is known to playa major role in influencing the form and dynamics of the hair follicle, which probably involves regulatory substances crossing the basal lamina. But little is known about the junctions between the dermal papilla and the surrounding epithelial cells of the hair bulb, or between the connective tissue and the epithelial cells on the outside of the hair follicle. This study was performed to identify the ultrastructural differences between dermoepidermal junction of the skin and connective tissue-epithelial junctions on the outside of the hair follicle and around the dermal papilla of normal anagen hair follicles in the human fetal scalp skin. Electron microscopic findings of dermoepidermal junction in scalp skin showed that basal lamina was very irregular and undulated, and it contained many attachment plaques of hemidesmosomes with sub-basal dense plates, tonofilaments, and anchoring filaments. Also invaginations of plasma membrane of basal keratinocytes were seen. There were clear differences both on the outside of the follicle and around the dermal papilla as compared with similar junction in the skin. In particular, neither hemidesmosomes nor tonofilaments, as seen in dermoepidermal junction, were observed in the dermal papilla. Also attachment plaque, sub-basal dense plate and anchoring filaments were not observed at the junction on the outside of the follicle and the dermal papilla. There were some differences between connective tissue-epithelial junctions on the outside of the hair follicle and around the dermal papilla, ie, smoothness of basal lamina and orthogonal arrangement of collagen fibers were seen in the outside of hair follicle, but not in the dermal papilla. These results indicate that the mechanical connection between the hair follicle and the connective tissue component is much weaker than that between the corresponding components in skin, and it reflects the dynamic processes during the anagen phase of the hair follicle compared to the relatively permanent state of the epidermis.

• Proceedings of the KOSOMBE Conference
• /
• 대한의용생체공학회 1992년도 춘계학술대회
• /
• pp.27-47
• /
• 1992

Engineers have developed new instruments that aid in diagnosis and therapy Ultrasonic imaging has provided a nondamaging method of imaging internal organs. A complex transducer emits ultrasonic waves at many angles and reconstructs a map of internal anatomy and also velocities of blood in vessels. Fast computed tomography permits reconstruction of the 3-dimensional anatomy and perfusion of the heart at 20-Hz rates. Positron emission tomography uses certain isotopes that produce positrons that react with electrons to simultaneously emit two gamma rays in opposite directions. It locates the region of origin by using a ring of discrete scintillation detectors, each in electronic coincidence with an opposing detector. In magnetic resonance imaging, the patient is placed in a very strong magnetic field. The precessing of the hydrogen atoms is perturbed by an interrogating field to yield two-dimensional images of soft tissue having exceptional clarity. As an alternative to radiology image processing, film archiving, and retrieval, picture archiving and communication systems (PACS) are being implemented. Images from computed radiography, magnetic resonance imaging (MRI), nuclear medicine, and ultrasound are digitized, transmitted, and stored in computers for retrieval at distributed work stations. In electrical impedance tomography, electrodes are placed around the thorax. 50-kHz current is injected between two electrodes and voltages are measured on all other electrodes. A computer processes the data to yield an image of the resistivity of a 2-dimensional slice of the thorax. During fetal monitoring, a corkscrew electrode is screwed into the fetal scalp to measure the fetal electrocardiogram. Correlations with uterine contractions yield information on the status of the fetus during delivery To measure cardiac output by thermodilution, cold saline is injected into the right atrium. A thermistor in the right pulmonary artery yields temperature measurements, from which we can calculate cardiac output. In impedance cardiography, we measure the changes in electrical impedance as the heart ejects blood into the arteries. Motion artifacts are large, so signal averaging is useful during monitoring. An intraarterial blood gas monitoring system permits monitoring in real time. Light is sent down optical fibers inserted into the radial artery, where it is absorbed by dyes, which reemit the light at a different wavelength. The emitted light travels up optical fibers where an external instrument determines O2, CO2, and pH. Therapeutic devices include the electrosurgical unit. A high-frequency electric arc is drawn between the knife and the tissue. The arc cuts and the heat coagulates, thus preventing blood loss. Hyperthermia has demonstrated antitumor effects in patients in whom all conventional modes of therapy have failed. Methods of raising tumor temperature include focused ultrasound, radio-frequency power through needles, or microwaves. When the heart stops pumping, we use the defibrillator to restore normal pumping. A brief, high-current pulse through the heart synchronizes all cardiac fibers to restore normal rhythm. When the cardiac rhythm is too slow, we implant the cardiac pacemaker. An electrode within the heart stimulates the cardiac muscle to contract at the normal rate. When the cardiac valves are narrowed or leak, we implant an artificial valve. Silicone rubber and Teflon are used for biocompatibility. Artificial hearts powered by pneumatic hoses have been implanted in humans. However, the quality of life gradually degrades, and death ensues. When kidney stones develop, lithotripsy is used. A spark creates a pressure wave, which is focused on the stone and fragments it. The pieces pass out normally. When kidneys fail, the blood is cleansed during hemodialysis. Urea passes through a porous membrane to a dialysate bath to lower its concentration in the blood. The blind are able to read by scanning the Optacon with their fingertips. A camera scans letters and converts them to an array of vibrating pins. The deaf are able to hear using a cochlear implant. A microphone detects sound and divides it into frequency bands. 22 electrodes within the cochlea stimulate the acoustic the acoustic nerve to provide sound patterns. For those who have lost muscle function in the limbs, researchers are implanting electrodes to stimulate the muscle. Sensors in the legs and arms feed back signals to a computer that coordinates the stimulators to provide limb motion. For those with high spinal cord injury, a puff and sip switch can control a computer and permit the disabled person operate the computer and communicate with the outside world.

• The Korean Journal of Physiology
• /
• 제24권1호
• /
• pp.27-37
• /
• 1990

The extrafetal transfer of $Li^{+}$ in amniotic fluid was studied in 45 pregnant rabbits. LiCl solution was administered either intravenously to mother or directly into the amniotic sac and monitored the appearance and disappearance of $Li^{+}$ in the amniotic fluid, then calculated the transfer rate of $Li^{+}$ of extrafetal origin. To study the transplacental $Li^{+}$ transfer, a solution of 150 mM LiCl was infused continuously via maternal vein (initial dose: 0.7 mmol/kg, maintaining dose: 0.03 mmol/kg/min) and the $Li^{+}$ concentration was measured in maternal blood and amniotic fluid after 60 and 120 minutes of infusion. Change in the volume of aminotic fluid was determined by Congo red dilution method at the same time. Effects of duration of gestation was not considered in this study. Extrafetal transport of $Li^{+}$ into the amniotic fluid was estimated by comparing the $Li^{+}$ concentration and volume of amniotic fluid determined before and after ligating the placental vessels. Extrafetal $Li^{+}$ transport from the amniotic fluid was determined by observing the time dependent disappearance of $Li^{+}$ and Congo red in amniotic fluid after injecting 0.5 ml solution of 15 mM or 90 mM LiCl and 50 mg/ml Congo red. Following are the results obtained: 1) During infusion of LiCl through maternal vein the ratio of the aminotic $Li^{+}$/maternal plasma $Li^{+}$ increased significantly along with the increment of fetal weight. 2) The volume of amniotic fluid of larger fetuses than 20.5 gm increased significantly during administration of LiCl while that of smaller fetuses did not change. 3) After umbilical cord ligation the $Li^{+}$ concentration of amniotic fluid of larger fetuses than 20.5 gm was decreased to $59.9{\pm}10.3%$ and $56.9{\pm}42.9%$ $(mean{\pm}S.D.)$ of those of control group after 60 and 120 minutes of LiCl infusion respectively. In amniotic fluid of smaller fetuses than 20.5 gm, there was no significant difference between control and ligation groups. 4) The disappearance rate of Congo red in the amniotic fluid was $45.2{\pm}8.2%/hr$. 5) The disappearance rate of $Li^{+}$ after intraamniotic injection of LiCl depended on the amount injected. On injecting $7.5\;{\mu}mol$ LiCl, $Li^{+}$ disappeared rapidly from the amniotic fluid and the rates after 60 min and 90 min were $97.0{\pm}2.8,\;98.5{\pm}2.0%$ respectively. On injecting $45\;{\mu}mol$ LiCl, the rates were $56.0{\pm}15.4,\;78.9{\pm}14.5%$ at 60 and 90 min. 6) From the above results it was concluded: a) $Li^{+}$ transfer into the amniotic fluid increased along with the fetal growth and one half of $Li^{+}$ influx is through the extrafetal route even after the maturation of fetal kidney. b) One half of the $Li^{+}$ transfer from the amniotic fluid was through swallowing of fetus, while the remaining half was transfered rapidly through amniotic membrane, which was concentration limited.

• Archives of Pharmacal Research
• /
• 제28권4호
• /
• pp.421-432
• /
• 2005

In order to understand the renal reabsorption mechanism of neutral amino acids via amino acid transporters, we have isolated human L-type amino acid transporter 2 (hLAT2) and human T-type amino acid transporter 1 (hTAT1) in human, then, we have examined and compared the gene structures, the functional characterizations and the localization in human kidney. Northern blot analysis showed that hLAT2 mRNA was expressed at high levels in the heart, brain, placenta, kidney, spleen, prostate, testis, ovary, lymph node and the fetal liver. The hTAT1 mRNA was detected at high levels in the heart, placenta, liver, skeletal muscle, kidney, pancreas, spleen, thymus and prostate. Immunohistochemical analysis on the human kidney revealed that the hLAT2 and hTAT1 proteins coexist in the basolateral membrane of the renal proximal tubules. The hLAT2 transports all neutral amino acids and hTAT1 transports aromatic amino acids. The basolateral location of the hLAT2 and hTAT1 proteins in the renal proximal tubule as well as the amino acid transport activity of hLAT2 and hTAT1 suggests that these transporters contribute to the renal reabsorption of neutral and aromatic amino acids in the basolateral domain of epithelial proximal tubule cells, respectively. Therefore, LAT2 and TAT1 play essential roles in the reabsorption of neutral amino acids from the epithelial cells to the blood stream in the kidney. Because LAT2 and TAT1 are essential to the efficient absorption of neutral amino acids from the kidney, their defects might be involved in the pathogenesis of disorders caused by a disruption in amino acid absorption such as blue diaper syndrome.

• Journal of Korean Neurosurgical Society
• /
• 제30권3호
• /
• pp.263-271
• /
• 2001

Objective : Glioblastomas, the most common type of primary brain tumors, are highly invasive and cause massive tissue destruction at both the tumor invading edges and in areas that are not in direct contact with glioma cells. As a result, patients with high-grade gliomas are faced with a poor prognosis. Such grim statistics emphasize the need to better understand the mechanisms that underlie glioma invasion, as these may lead to the identification of novel targets in the therapy of high grade gliomas. Protein kinase C(PKC) is a family of serine/threonine kinases and an important signal transduction enzyme that conveys signals generated by ligand-receptor interaction at the cell surface to the nucleus. PKC appears to be critical in regulating many aspects of glioma biology. The purpose of this study was to assess accurately the role of PKC in the invasion regulation of human gliomas based on hypothesis that protein kinase C(PKC) is functional in the process of glial tumor cell invasion. Method : To test this hypothesis, U-87 malignant glioma cell line intracellular PKC levels were up and down regulated and their invasiveness was tested. Intracellular PKC level was characterized using PKC activity assays. Invasion assays including barrier migration and spheroid confrontation were used to study the relationship between PKC concentration and invasiveness. Result : The cell line which were treated by PKC inhibitor tamoxifen and hypericin exhibited decreased PKC activity and decreased invasive abilities dose dependently both in matrigel invasion assay and tumor spheroid fetal rat brain aggregates(FRBA) confrontation assay. However, the cell line that was treated by PKC activator 12-O-tetradecanylphorbol-13acetate(TPA) did not exhibit increases in either PKC activity or invasive ability. Conclusion : These studies suggest that PKC may be a useful molecular target for the chemotherapy of glioblastoma and other malignancies and that a therapeutic approach based on the ability of PKC inhibitors may be helpful in preventing invasion.

• The Korean Journal of Physiology
• /
• 제27권1호
• /
• pp.13-25
• /
• 1993

To study the regulation of amniotic fluid volume and electrolyte concentration by the Membranes surrounding the amniotic fluid, the rate of $Li^+$ disappearance from amniotic sac of expired fetuses were examined while increasing the amniotic volume and osmolarity in rabbits. After intraamniotic injection of 1 ml isosmotic saline (about 20% of the amniotic fluid volume) containing 15 mM LiCl and 0.5 g/L Censored, the time courses of $Li^+$ and Censored disappearance were determined. From there the $Li^+$ clearance through the extrafetal routes was estimated and compared with that obtained from living fetuses. The volume, $Na^+$ concentration and osmolarity of amniotic fluid were measured and their relationships with $Li^+$ disappearance were evaluated. The fellowing results were obtained: 1. The rate of disappearance from amniotic fluid of living fetuses during the first 30 minutes was strikingly higher for $Li^+$ than for Censored, suggesting that extrafetal routes exist. At 60 and 90 minutes, however, the disappearance rate of $Li^+$ was less than that of Censored, suggesting the possibility of $Li^+$ reentry through fetal urination. 2. The disappearance of $Li^+$ from the amniotic fluid of the expired fetus was substantial, although lower than that of living fetuses, throughout the experimental period. 3. The $Na^+$ concentration and the osmolarity of the amniotic fluid of expired fetus measured 30 minutes after an intraamniotic injection of isoosmotic saline showed wide variation, but thereafter they changed gradually towards the normal extracellular fluid level. 4. When the amniotic fluid was iso- or hyposmolar, the rate of $Li^+$ disappearance from the amniotic fluid of the expired fetuses showed little variation. However, when the amniotic fluid was hyperosmolar, the rate at 30 minutes was markedly lower than those of isosmotic or hyposmotic amniotic fluid. At 90 minutes, the rate of $Li^+$ disappearance in hyperosmolar fluid reached a similar level to the rate in isosmolar fluid. 5. The intraamniotic injection of 400 mOsm/L saline solution decreased the disappearance rate of $Li^+$ from expired fetuses, while the injection of mannitol into the maternal vein induced no significant change. From these results it is concluded that: 1) a significant amount of $Li^+$ may leave the amniotic fluid via filtration through the membranes surrounding the amniotic fluid, 2) during hyperosmolar challenge to amniotic fluid, osmotic bulk flow might counteract the filterable loss, and 3) $Li^+$ disappearance might continue even after the volume and osmolarity of the amniotic fluid have recovered to control values.

• Applied Microscopy
• /
• 제23권1호
• /
• pp.56-76
• /
• 1993

To investigate the histogenesis of tracheal epithelium in Sprague-Dawley strain rat, the author has used the fetal rats at the 16th, 18th, 20th and 22nd prenatal day and neonatal rats at the 1st and 7th day as well as rats at age of 5, 10 and 15 weeks after birth as experimental animals. Specimens were double stained with uranyl acetate and lead citrate for electron microscopic study. The results obtained were as follows; 1. At the 16th day of gestational age, ciliated cells were found in tracheal epithelium and light and dark ciliated cells possessing numerous mitochondria and smooth endoplasmic reticulum in the cytoplasm at the 22nd day of gestational age of the rat are observed. 2. At the 16th day of gestataional age, basal cells lying upon the basement membrane and having large numbers of glycogen particles in the cytoplasm, were found and at the 22nd day of gestational age, basal cells possessing numerous polysomes in the cytoplasm were observed. 3. At the 20th gestational age of the rat, microvillous cells possessing many rough endoplasmic reticulum and mitochondria as well as microvilli protruding into the lumen were found in tracheal epithelium. 4. At the 5th week after birth brush cell having profound filamentous strands and many pinocytic vesicles in the cytoplasm, was visible in the tracheal epithelium. 5. At the 15th week after birth large proportions of tracheal epithelium were lined with ciliated cells. Cosequently it is suggested that pseudostratified ciliated columnar epithelium was differentiated at the 16th day of gestational age, in addition cytoplasmic organelles of the microvillous and basal cells were matured at the 20th and 22nd gestational age, respectively and most of the part of the tracheal epithelium was lined with ciliated cells at the 15th week after birth.

• Asian Journal of Turfgrass Science
• /
• 제25권2호
• /
• pp.133-137
• /
• 2011

Winterkill can be defined as any injury including freeze stress kill, winter desiccation, and low temperature disease to turfgrass plants that occurs during the winter period. The major damages from winterkill were low temperature kill, crown hydration, and winter desiccation. Low temperature kill is caused by air and soil temperature. Soil temperature affect more severe to turfgrass than air temperature because low soil temperature cause fetal damage to turfgrass crown. Crown hydration is a form of winter injury in which intercellular water within the plant freezes and causes physical injury to the cell membrane and wall. This is eventually resulted in dehydration of cell. Winter desiccation is the death of leaves or whole plants due to drought during the winter period. To reduce winterkill damage, cultivar selection is very important. If changing cultivar is not allowed, cold temperature hardiness needs to be increased by providing nutrients especially phosphorus and potassium in the late fall. Turf cover is effective way to reduce winterkill damage. Remaining snow is positive process to reduce winterkill damage by insulating soil temperature. The previous researches reported many materials as turf cover such as straw, polypropylene, polyester, and wood mat. Aeration and topdressing is one of the process against winterkill. Both methods are mainly conducted to reduce thickness of thatch layer. In recent, relatively new materials called black or winter topdressing sand are used to protect soil temperature from low air temperature and thaw ice crystal that may remain in soil.

• The Korean Journal of Cytopathology
• /
• 제6권2호
• /
• pp.106-115
• /
• 1995

Reactive humsn mesothelial cells were examined by immunocytochemical stain with intermediate filaments (cytokeratin [CK1, CK7, CK8, CK18, CD19), vimentin, desmin, actin), epithelial membrane antigen, carcinoembryonic antigen (CEA), MHC class II antigen (HLA-DR), LeuM-1 (CD15), $\alpha1-antitrypsin$(ACT), $\alpha1-antichymotrypsin$ (ACHT), CD68(KP-1) and FcyRIII(CD16). The mesothelial cells were isolated from patients with liver cirrhosis and pleural effusion, and short-term cultured in RPMI 1640 media containing 10% heat inactivated fetal calf serum and 1% identical supernatant fluid of the patients' transudates. The results obtained are as follows 1. The cultured-reactive mesothelial cells were positive for the protein of cytoskeleton such as cytokeratin and vimentin, but negative for desmin and actin. The resting mesothelial cells showed positive reactions for cylokeratin, but negative for vimentin, desmin and actin. 2. The primary antibodies to the cytokeratin were strongly reactive for CK1, CK8 and CK18 but negative for CK7 and CK19 in both reactive and resting mesothelial cells. 3. Resting mesothelial cells showed negative reactions for CEA, but strong positive reactions in cultured-reactive mesothelial cells. 4. The markers for the monocytes/histiocytes(CD11b, CD14, CD16, CD68, Iysozyme and $\alpha1-antitrypsin$ and $\alpha1-antichymotrypsin$) were nonreactive in resting mesothelial cells, but lysozyme and $\alpha1-antitrypsin$ were weakly reactive in reactive and proliferative mesothelial cells. 5. MHC Class II molecule(HLA-DR antigen) was negative in both resting and reactive mesothelial cells. These results suggest that the short-term cultured, reactive mesothelial cells show a newly aberrant expression of the vimentin and calcine-embryonic antigen. The reason of the aberrant expression of the intermediate filament and oncofetal antigen in reactive and proliferative mesothelial cells should be further evaluated.