This study was undertaken to compare the activity of masticatory muscle between normal occlusion and anterior openbite. 33 subjects without the experience of orthodontic treatment, missing teeth and the symptom of T. M. disorders were selected for this study: 25 subjects were normal occlusion and 8 subjects were anterior openbite. The ten items were measured from the cephalometric headplates, and EMG recordings of the anterior temporal and masseter muscle were taken at rest position and during maximum clenching at centric occlusion. All data were analyzed and processed with the computer statistical method. The following results were obtained: 1. At rest position, the muscle activities of both temporal and masseter muscle were higher in anterior openbite than in normal occlusion. 2. During maximum clenching, the muscle activities of both temporal and masseter muscle were prominently lower in anterior openbite than in normal occlusion. 3. At rest position, the temporal muscle of anterior openbite showed the highest muscle activity, but showed the lowest muscle activity during maximum clenching. 4. Anterior openbite showed closer interrelationship between facial morphology and the muscle activity, and the muscle activity was more influenced by the form of mandible than that of maxilla.
Mandible ; Masseter Muscle ; Masticatory Muscles ; Maxilla ; Open Bite* ; Temporal Muscle ; Tooth

Nitric oxide(NO) has been known to play an important role as a signal molecule in many parts of the organism as well as a cytotoxic effector molecule of the nonspecific immune response. Excessive NO has been reported to exert cytotoxic effect by direct toxicity or by reacting to superoxide radicals during ischemia-reperfusion. Therefore is strongly needed a study directly measuring NO release designed to better clarify roles of NO in ischemia-reperfusion injury and its mechanism. Male Sprague-Dawley rats were anesthetized with urethane(1g/kg) intraperitoneally, and pedicled gastrocnemius muscle flaps were elevated. The elevated flaps were subjected to 4 hours of arterial ischemia and then reperfused for 2 hours. And then NO current was measured in the gastrocnemius muscle using NO-selective microelectrode system. NO release gradually increased and then decreased in the rat gastrocnemius muscle during both ischemic and reperfusion period. The average NO releases from baseline during ischemia and reperfusion were 10405 +/- 2663 and 2513 +/- 970 picoamperes(pA), respectively. The ischemia-reperfusion caused substantial histological damage in the skeletal muscle, in which a profusion of red blood cells was observed due to extravasation of vessels, rupturing of microcirculation, and leukocyte infiltration, compared to the damage in control and ischemic rat gastrocnemius muscles. From the above results, ischemia-reperfusion injury was developed more severely during reperfusion than ischemia, and NO increased during ischemia and reperfusion as a biphasic pattern in the rat gastrocnemius muscle.
Animals ; Erythrocytes ; Humans ; Ischemia ; Leukocytes ; Male ; Microcirculation ; Microelectrodes ; Muscle, Skeletal* ; Muscles ; Nitric Oxide* ; Rats* ; Rats, Sprague-Dawley ; Reperfusion ; Reperfusion Injury ; Superoxides

PURPOSE: Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein involved in DNA repair and redox modulation. Recently, serum and urinary APE1/Ref-1 levels were reported to be increased in patients with bladder cancer. Genetic variations of APE/Ref-1 are associated with the risk of cancer. However, the effect of APE1/Ref-1 variants on its secretory activity is yet unknown. METHODS: APE1/Ref-1 variants were evaluated by DNA sequencing analysis of reverse transcription polymerase chain reaction products in coding DNA sequences (CDS) of APE1/Ref-1 in bladder tissue samples from patients with bladder cancer (n=10). Secretory activity of APE1/Ref-1 variants was evaluated with immunoblot and enzyme-linked immunosorbent assay of the culture medium supernatants. RESULTS: Four different substitution mutants (D148E, I64V/D148E, W67R/D148E, and E86G/D148E) of APE1/Ref-1 were identified in bladder cancer specimens. However, deletion mutants of APE1/Ref-1 CDS were not found. The secretory activity of the APE1/Ref-1 variants (D148E, I64V/D148E, and E86G/D148E) was increased compared to that of wild type APE1/Ref-1. Furthermore, the secretory activity in basal or hyperacetylated conditions was much higher than that in APE1/Ref-1 D148E-transfected HEK293 cells. CONCLUSIONS: Taken together, our data suggest that the increased secretory activity of D148E might contribute to increased serum levels of APE1/Ref-1 in patients with bladder cancer.
Base Sequence ; Clinical Coding ; DNA Repair ; Enzyme-Linked Immunosorbent Assay ; Genetic Variation ; HEK293 Cells ; Humans* ; Oxidation-Reduction ; Point Mutation ; Polymerase Chain Reaction ; Reverse Transcription ; Sequence Analysis, DNA ; Urinary Bladder Neoplasms* ; Urinary Bladder*

Caesalpinia sappan (C. sappan) is a medicinal plant used for promoting blood circulation and removing stasis. During a screening procedure on medicinal plants, the ethylacetate extract of the lignum of C. sappan (CLE) showed inhibitory activity on arginase which has recently been reported as a novel therapeutic target for the treatment of cardiovascular diseases such as atherosclerosis. CLE inhibited arginase II activity prepared from kidney lysate in a dose-dependent manner. In HUVECs, inhibition of arginase activity by CLE reciprocally increased NOx production through enhancement of eNOS dimer stability without any significant changes in the protein levels of eNOS and arginase II expression. Furthermore, CLE-dependent arginase inhibition resulted in increase of NO generation and decrease of superoxide production on endothelium of isolated mice aorta. These results indicate that CLE augments NO production on endothelium through inhibition of arginase activity, and may imply their usefulness for the treatment of cardiovascular diseases associated with endothelial dysfunction.
Animals ; Aorta ; Arginase ; Atherosclerosis ; Blood Circulation ; Caesalpinia ; Cardiovascular Diseases ; Endothelium ; Kidney ; Mass Screening ; Mice ; Nitric Oxide ; Nitric Oxide Synthase Type III ; Plants, Medicinal ; Superoxides

The effects of pH on K+ currents were investigated in single smooth muscle cells isolated from the thoracic aorta of Wistar-Kyoto rats. Whole-cell K+ currents were recorded in the conventional configuration of the voltage-clamp technique. Pinacidil (10muM) activated the whole-cell current and the pinacidil-activated current was completely inhibited by glibenclamide (10muM), an inhibitor of ATP-sensitive K+ channel (KATP channel). Pinacidil-activated current was reversed at near the K+ equilibrium potential. This current was time- and voltage-independent and reduced by elevating intracellular ATP. Pinacidil-activated current was reduced by lowering the external pH. However, alteration of internal pH has controversial effects on pinacidil-activated current. When the single cell was dialyzed with 0.1 mM ATP, alteration of internal pH had no effect on pinacidil-activated K+ current. In the contrast, when the single cell was dialyzed with 3 mM ATP, pinacidil-activated current was increased by lowering internal pH. Our results suggest that K+ channel activated by pinacidil may be KATP channel and internal H+ may reduce the inhibitory effect of ATP on KATP channel.
Adenosine Triphosphate ; Animals ; Aorta, Thoracic ; Glyburide ; Hydrogen-Ion Concentration* ; Muscle, Smooth* ; Muscle, Smooth, Vascular ; Myocytes, Smooth Muscle* ; Patch-Clamp Techniques ; Pinacidil ; Rats*

Using the patch-clamp technique, we investigated the alteration of 4-aminopyridine(4-AP)-sensitive, voltage-dependent K+ channel (KV) in the mesenteric arterial smooth muscle cell (MASMC) of renovascular hypertensive model, one-kidney one-clip Goldblatt hypertensive rat (GBH). To isolate KV current, internal pipette solution contained 5 mM ATP and 10 mM EGTA. Under these condition, MASMC was depolarized by 4-AP, but charybdotoxin did not affect membrane potential. Membrane potential of hypertensive cell (- 40.3 +/- 3.2 mV) was reduced when compared to that of normotensive cell (-59.5 +/- 2.8 mV). Outward K+ current of hypertensive cell was significantly reduced when compared to normotensive cell. At 60 mV, the outward currents were 19.10 +/- 1.91 and 14.06 +/- 1.05 pA/pF in normotensive cell and hypertensive cell respectively. 4-AP-sensitive K+ current was also smaller in hypertensive cell (4.28 +/- 0.38 pA/pF) than in normotensive cell (7.65 +/- 0.52 pA/pF). The values of half activation voltage (V1/2) and slope factor (k1) as well as the values of half inactivation voltage (V1/2) and slope factor (k1) were virtually similar between GBH and NTR. These results suggest that the decrease of 4-AP-sensitive K+ current contributes to a depolarization of membrane potential, which leads to development of vascular tone in GBH.
Adenosine Triphosphate ; Animals ; Charybdotoxin ; Egtazic Acid ; Membrane Potentials ; Muscle, Smooth* ; Myocytes, Smooth Muscle* ; Patch-Clamp Techniques ; Rats*

Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein that plays a central role in the cellular response to DNA damage and redox regulation against oxidative stress. APE1/Ref-1 functions in the DNA base excision repair pathway, the redox regulation of several transcription factors, and the control of intracellular redox status through the inhibition of reactive oxygen species (ROS) production. APE1/Ref-1 is predominantly localized in the nucleus; however, its subcellular localization is dynamically regulated and it may be found in the mitochondria or elsewhere in the cytoplasm. Studies have identified a nuclear localization signal and a mitochondrial target sequence in APE1/Ref-1, as well as the involvement of the nuclear export system, as determinants of APE1/Ref-1 subcellular distribution. Recently, it was shown that APE1/Ref-1 is secreted in response to hyperacetylation at specific lysine residues. Additionally, post-translational modifications such as phosphorylation, S-nitrosation, and ubiquitination appear to play a role in fine-tuning the activities and subcellular localization of APE1/Ref-1. In this review, we will introduce the multifunctional role of APE1/Ref-1 and its potential usefulness as a therapeutic target in cancer and cardiovascular disease.
Active Transport, Cell Nucleus ; Biomarkers ; Cardiovascular Diseases ; Cytoplasm ; DNA ; DNA Damage ; DNA Repair ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; Lysine ; Mitochondria ; Nuclear Localization Signals ; Oxidation-Reduction ; Oxidative Stress ; Phosphorylation ; Protein Processing, Post-Translational ; Reactive Oxygen Species ; Transcription Factors ; Ubiquitin ; Ubiquitination

PURPOSE: Peroxynitrite plays a critical role in vascular pathophysiology by increasing arginase activity and decreasing endothelial nitric oxide synthase (eNOS) activity. Therefore, the aims of this study were to investigate whether arginase inhibition and L-arginine supplement could restore peroxynitrite-induced endothelial dysfunction and determine the involved mechanism. MATERIALS AND METHODS: Human umbilical vein endothelial cells (HUVECs) were treated with SIN-1, a peroxynitrite generator, and arginase activity, nitrite/nitrate production, and expression levels of proteins were measured. eNOS activation was evaluated via Western blot and dimer blot analysis. We also tested nitric oxide (NO) and reactive oxygen species (ROS) production and performed a vascular tension assay. RESULTS: SIN-1 treatment increased arginase activity in a time- and dose-dependent manner and reciprocally decreased nitrite/nitrate production that was prevented by peroxynitrite scavenger in HUVECs. Furthermore, SIN-1 induced an increase in the expression level of arginase I and II, though not in eNOS protein. The decreased eNOS phosphorylation at Ser1177 and the increased at Thr495 by SIN-1 were restored with arginase inhibitor and L-arginine. The changed eNOS phosphorylation was consistent in the stability of eNOS dimers. SIN-1 decreased NO production and increased ROS generation in the aortic endothelium, all of which was reversed by arginase inhibitor or L-arginine. N(G)-Nitro-L-arginine methyl ester (L-NAME) prevented SIN-1-induced ROS generation. In the vascular tension assay, SIN-1 enhanced vasoconstrictor responses to U46619 and attenuated vasorelaxant responses to acetylcholine that were reversed by arginase inhibition. CONCLUSION: These findings may explain the beneficial effect of arginase inhibition and L-arginine supplement on endothelial dysfunction under redox imbalance-dependent pathophysiological conditions.
15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid ; Acetylcholine ; Arginase* ; Arginine ; Blotting, Western ; Endothelium ; Human Umbilical Vein Endothelial Cells ; NG-Nitroarginine Methyl Ester ; Nitric Oxide ; Nitric Oxide Synthase Type III* ; Oxidation-Reduction ; Peroxynitrous Acid ; Phosphorylation* ; Reactive Oxygen Species

PURPOSE: The purpose of this study is to understand and analyse the actual education environment of the subject, life science, and how it is taught in high school, and thereby make a realistic assessment of whether a medical life science experience program is necessary. METHODS: The test method to assess the actual curriculum was developed by Life Science teachers, medical school faculty and education specialists. The subject was divided into 4 areas consisting of 6 items each. Our survey was mailed out. The analysis consisted of frequency analysis, chi-square analysis, correlation analysis, and variance analysis using SPSS 13.0 for Windows. RESULTS: Over 90% of Life Science class teachers agreed that teaching should be done through lectures and lab experiments in parallel. However, currently the class is heavily lecture-oriented due to the lack of lab facilities, the lack of budget and the difficulty of organizing lab courses. Due to the nature of the subject, lab experiments are crucial. Therefore, it is recommended that a biomedical science experience program be included in the curriculum. This program should be offered during vacation and geared toward high school freshmen and sophomore students. CONCLUSION: This research clearly showed the need to develop a high school biomedical science experience program. In order for the program to be successful, one must take into consideration the safety of experiments, the capability of the instructors, the development of a variety of experiments, the accessibility of the location of the lab, securing interest in education at a community level and the compilation of an experience program at every educational level.
Biological Science Disciplines* ; Budgets ; Curriculum ; Education* ; Humans ; Lectures ; Postal Service ; Schools, Medical ; Specialization

Elevated plasma concentration of native low-density lipoprotein (nLDL) is associated with vascular smooth muscle cell (VSMC) activation and cardiovascular disease. We investigated the mechanisms of superoxide generation and its contribution to pathophysiological cell proliferation in response to nLDL stimulation. Lucigenin-induced chemiluminescence was used to measure nLDL-induced superoxide production in human aortic smooth muscle cells (hAoSMCs). Superoxide production was increased by nicotinamide adenine dinucleotide phosphate (NADPH) and decreased by NADPH oxidase inhibitors in nLDL-stimulated hAoSMC and hAoSMC homogenates, as well as in prepared membrane fractions. Extracellular signal-regulated kinase 1/2 (Erk1/2), protein kinase C-theta (PKCtheta) and protein kinase C-beta (PKCbeta) were phosphorylated and maximally activated within 3 min of nLDL stimulation. Phosphorylated Erk1/2 mitogen-activated protein kinase, PKCtheta and PKCbeta stimulated interactions between p47phox and p22phox; these interactions were prevented by MEK and PKC inhibitors (PD98059 and calphostin C, respectively). These inhibitors decreased nLDL-dependent superoxide production and blocked translocation of p47phox to the membrane, as shown by epifluorescence imaging and cellular fractionation experiments. Proliferation assays showed that a small interfering RNA against p47phox, as well as superoxide scavenger and NADPH oxidase inhibitors, blocked nLDL-induced hAoSMC proliferation. The nLDL stimulation in deendothelialized aortic rings from C57BL/6J mice increased dihydroethidine fluorescence and induced p47phox translocation that was blocked by PD98059 or calphostin C. Isolated aortic SMCs from p47phox-/- mice (mAoSMCs) did not respond to nLDL stimulation. Furthermore, NADPH oxidase 1 (Nox1) was responsible for superoxide generation and cell proliferation in nLDL-stimulated hAoSMCs. These data demonstrated that NADPH oxidase activation contributed to cell proliferation in nLDL-stimulated hAoSMCs.
Animals ; Aorta/*cytology ; Cell Line ; Cell Proliferation ; Cells, Cultured ; Humans ; Lipoproteins, LDL/*metabolism ; Mice, Inbred C57BL ; Mitogen-Activated Protein Kinases/metabolism ; Muscle, Smooth, Vascular/cytology ; Myocytes, Smooth Muscle/*cytology ; NADPH Oxidase/*metabolism ; Phosphorylation ; Protein Kinase C/metabolism ; Signal Transduction ; Superoxides/metabolism