BACKGROUND: Reactive oxygen species (ROS) induce lipid peroxidation and tissue damage in the isolated rabbit thoracic aorta. The aim of this study was to explore the influence of the propofol and midazolam against ROS in the isolated rabbit thoracic aortic endothelium. METHODS: Eighteen white male rabbits (weighing 2.0-2.5 kg) were used. The thoracic aorta was dissected free and cut into rings (3-4 mm) and then suspended in a organ bath filled with 10 ml Krebs solution bubbled with 5% CO2 95% O2 at 37 degrees C. Aortic rings were then equilibrated for 90 min, and a resting tension of 1.5 g was applied. The Krebs solution was changed every 15 min. Isometric tension was recorded with transducer coupled to a data acqusition system (Biopac Inc. USA) on a PC. After precontraction with norepinephrine (NE, 10(-6)M), changes in tension were measured following the cumulative administration of acetylcholine (ACh 3x10(-7), 10(-6) and 3x10(-6)M) and nitroglycerin (NTG, 10(-5)M). Data are expressed as percentage of the 10 5 M NTG-induced relaxation (ACh/NTG). The ACh/NTG, before and after electrolysis were defined as the control and the experimental groups. The aortic rings were pretreated with propofol (3x10(-5), 10(-4), 3x10(-4) and 5.7x10(-4) M, n = 8, 10, 15, 13), midazolam (10(-4)M, n = 7), catalase (1,000 U/ml, n = 12), mannitol (3x10(-4)M, n = 5) or not pretreated group (Free, n = 6). After 30 minutes, the aortic rings were exposed to ROS generated by electrolysis (DC 9 V, 20 mA, aortic rings 1 cm away from electrode) in Krebs solution for 2 minutes, which was then changed for physiologic buffered salt solution. The aortic rings were precontracted with NE and vasorelaxation was induced with ACh and NTG at the above mentioned concentrations. RESULTS: Propofol produced vasorelaxation of NE-precontracted thoracic aorta in a dose-dependent fashion in all groups of propofol (3x10(-5), 10(-4), 3x10(-4) and 5.7x10(-4)M) even after ROS attack (P < 0.05 vs control value). Catalase produced vasorelaxation after ROS attack (P < 0.05 vs control value).On the other hand, ACh-induced significant endothelium-dependent vasorelaxation were not observed in the midazolam or mannitol pretreated group or the non-pretreated group (P <0.05 vs control group). CONCLUSIONS: These findings suggest that propofol and catalase preserve ACh induced endothelium-dependent vasorelaxation and that propofol has a concentration dependent ROS scavenging effect like catalase.
Acetylcholine ; Aorta, Thoracic ; Baths ; Catalase ; Electrolysis ; Endothelium ; Hand ; Humans ; Lipid Peroxidation ; Male ; Mannitol ; Midazolam ; Nitroglycerin ; Norepinephrine ; Propofol* ; Rabbits ; Reactive Oxygen Species* ; Relaxation ; Transducers ; Vasodilation

BACKGROUND: Intracellular ionized calcium plays a central role in the transduction of external stimuli as a critical second messenger. The spectral properties of fluo-3 allows the analysis of intracellular ionized calcium level by flow cytometers. The aim of this study is to assess the performance of flow cytometer for measuring intracellular ionized calcium level using fluo-3 and to define the reference interval of intracellular ionized calcium level of lymphocytes from healthy people, and to find out the clinical implications according to various disorders. METHODS: For the analytical performance of flow cytometer on determining the concentration of intracellular ionized calcium, precision study, lowest limit of detection, analytical range, and the loading stability of fluo-3 were per foamed. Fifty-four cases of healthy people, 52 cases of renal transplant patients, and 20 cases of diabetes mellitus patients were included in this study. RESULTS: Loading effect of fluo-3 at room temperature was stable upto 5 hours. Lowest limit of detection of ionized calcium concentration was 4.34 nM at in-situ calibration procedure. Within-run and among-day intraindividual CVs of in-situ calibration procedure were 6.67% and 13.99% respectively, and of optical calibration procedure were 13.86% and 16.12% respectively. The reference interval of cytosolic free calcium level for healthy people ranged 73.54 - 155.09 nM without sexual differences. The level of intracellular ionized calcium was lowered by 36.9% on renal transplant group in comparison with healthy control group. But, level of cytosolic free calcium was Increased upto 276.0% on acute rejection group and 159.1% on diabetes mellitus group compared to control group. CONCLUSIONS: These results reveal that in-situ calibration method for intra cellular ionized calcium using flow cytometry with flue-3 can be regarded as an accurate and standardized method. Quantitation of intracellular ionized calcium level might be used as the monitoring test for early detection of acute rejection after renal transplantation.
Calcium* ; Calibration ; Cytosol ; Diabetes Mellitus ; Flow Cytometry* ; Humans* ; Kidney Transplantation ; Limit of Detection ; Lymphocytes* ; Second Messenger Systems

BACKGROUND: Morphine has a direct action on morphine receptors in the brain and spinal cord. Intrathecally administered L-NAME, a nitric oxide synthase inhibitor, is known to have an antinociceptive effect on formalin-induced pain in animal studies. Efficacy of intrathecally administered ketorolac, a cyclooxygenase inhibitor, is somewhat controversial. The interactions of intrathecally administered morphine, ketorolac and L-NAME on formalin-induced nociception was studied. METHODS: Male Sprague-Dawley rats were implanted with chronic lumbar intrathecal catheters and were tested for paw flinch by a formalin injection. Drugs were intrathecally administered 15 min before the formalin injection, and biphasic painful behaviors were observed. We obtained the ED50 for each agent (ketorolac, L-NAME and morphine). ED50 fractions (1, 1/2 and 1/4) of drug combinations of L-NAME-ketorolac, morphine-L-NAME and ketorolac-morphine were administered. The ED50 of each combined drug was established and isobolographic analysis of the drug interactions was carried out. RESULTS: Intrathecal administration of ketorolac, L-NAME and morphine produced a dose-dependent suppression of pain behaviors in phase 2. ED50 values were 297.04micro gram for ketorolac, 207.46micro gram for L-NAME and 0.17micro gram for morphine in phase 2. Isobolographic analysis showed that the combination of intrathecal morphine and L-NAME synergistically reduced pain behaviors in phase 2. CONCLUSIONS: Intrathecally administered morphine, L-NAME and ketorolac produced a dose-dependent decrease in the number of paw flinches in both phase 1 and phase 2 on the formalin test. Morphine with L-NAME showed synergistic analgesic effects on formalin-induced pain in phase 2.
Animals ; Brain ; Catheters ; Drug Combinations ; Drug Interactions ; Formaldehyde ; Humans ; Ketorolac* ; Male ; Morphine* ; NG-Nitroarginine Methyl Ester* ; Nitric Oxide Synthase ; Nociception ; Pain Measurement ; Prostaglandin-Endoperoxide Synthases ; Rats* ; Rats, Sprague-Dawley ; Receptors, Opioid, mu ; Spinal Cord

OBJECTIVE: A number of disease-modifying anti-rheumatic drugs (DMARDs) have been shown to be more effective than placebo in the management of rheumatoid arthritis (RA). However, most course of DMARDs, except methotrexate, are discontinued after 2 or 3 years, because of toxicity, lack of efficacy or escape from control. The multi-drug resistance (MDR) is a phenomenon in which cells develop cross-resistance to many agents such as anthracyclin, vinca alkaloids and colchicine. In our hypothesis, MDR phenomenon could be implicated in acquired resistance to DMARDs in RA. We have established a mdr1 cell line and tested whether DMARDs are substrate for P-glycoprotein (P-gp). METHODS: The mdr1-cDNA was cloned into retroviral vector, and the recombinant retroviral vector was transfected into PA317 cells. The target cells, NIH3T3, were infected with recombinant retroviruses. A colony most resistant to vinblastin was selected for the following experiments; expression of mdr1 gene in NIH3T3 cells was confirmed by RT-PCR, and biological function of mdr1 gene product, P-gp, was tested using Rhodamine-123 (Rh123) efflux assay. Resistance of the target cells expression P-gp which can survive against hydroxychloroquine (HCQ) and methotrxate (MTX) were measured by MTT assay. RESULTS: RT-PCR for mdr1 gene showed successful transfer of the gene into the NIH3T3 cells. Rh123 assay revealed expression of P-gp on the selected cells as follows; Rh123 efflux activity of uninfected NIH3T3 cells was 6%, that of PLXSN was 0.2%, and that of selected cells was 44%. The 50% proliferation inhibitory capacity of the selected cells were twice for HCQ but there was no difference of that for MTX. CONCLUSION: We established a mdr1 cell line and using the cell line, HCQ was a substrate of MDR, but MTX was not related to MDR.
Antirheumatic Agents* ; Arthritis, Rheumatoid ; Cell Line ; Clone Cells ; Colchicine ; Drug Resistance, Multiple ; Hydroxychloroquine ; Methotrexate ; P-Glycoprotein ; Retroviridae ; United Nations ; Vinca Alkaloids ; Zidovudine

Purpose: This study aimed to describe the desperate situation where the clinician should make decisions to further manage patients having experienced adverse drug reaction (ADR) to lamotrigine that is indicated to not easily controlled neuropsychiatric diseases. Methods: A descriptive analysis was done by thoroughly reviewing medical records of patients who were reported to have ADR to lamotrigine in a regional drug-safety center between 2010 and 2018. Results: Eighty-four cases of lamotrigine-related ADRs occurred in 80 patients. Skin lesions were most commonly observed in 70 cases (83.3%) and 14 cases (16.7%) had severe ADRs. Sixty-three subjects (78.8%) discontinued lamotrigine, while 17 (21.3%) continued it.At the time of discontinuation, 30.0% were prescribed aromatic antiepileptic drugs. Among 4 subjects who were eventually prescribed lamotrigine again after a period of discontinuation, 3 (75.0%) experienced its recurrence. Among patients who had taken alternative medications, the incidence of ADRs was higher in those being prescribed aromatic antiepileptic drugs than in the others being prescribed other than aromatic antiepileptic drugs (P = 0.013). Regarding the control of underlying diseases, as many as 65 (86.7%) and 68 (90.7%) failed to reach maintaining the resolved state from 6 months and 12 months after the substitution, respectively. Conclusion Patients can be easily trapped between the recurrence of ADRs and the treatment failure to a certain drug like lamotrigine, in which we can hardly find a reasonable alternative to manage them.

Ketamine hydrochloride is a phencyclidine derivatives and dissociative anesthetics. Ketamine induce the pulmonary vasoconetrietion in vivo. This study was designed to deter- mine the direct effect of the ketamine on the rabbit pulmonary artery in vitro. Isolated pulmonary artery was precontracted with norepinephrine (NE) 10-7M in the 20 ml organ bath. Concentration of ketamine was gradually increased 10-5M, 10 4M and 10-3M at 10 minutes intervals. I divided forty three experimental speeimens into 5 groups : pulmonary artery with and without endothelium, pretreated with indomethacin, nitrow-L-arginine methyl ester(L-NAME) and methylene blue. The results were as follows : 1. Norepinephrine precontracted pulmonary arterial tone wss significantly decreased by ketamine(10-3M), and the relaxing percent were 81.0 19.3, 60.6 55.4 (Mean S.D.) in unrubbed and rubbed endothelium, respectively (p<0.05). 2. The changes of vascular tone in denuding and intact groups were not significantly different. 3. Vasorelaxation induced by ketamine was not related with nitric oxide(NO) synthase, cyclooxygenase and soluble guanylate cyclase. Ketamine induce relaxation of the rabbit pulmonary artery, especially at 10-3 M concentra- tion. The relaxing effect was not related with endothelium presence, nitric oxide synthase, cyclooxygenase and soluble guanylate cyclase pathways. This data suggest that the relaxing effect of ketamine was not associated with endothelium, Nitric oxide, prostacyclin and cyclic guanosinemonophosphate.
Anesthetics, Dissociative ; Arteries ; Baths ; Endothelium ; Epoprostenol ; Guanylate Cyclase ; Indomethacin ; Ketamine* ; Methylene Blue ; NG-Nitroarginine Methyl Ester ; Nitric Oxide ; Nitric Oxide Synthase ; Norepinephrine ; Phencyclidine ; Prostaglandin-Endoperoxide Synthases ; Pulmonary Artery ; Relaxation ; Vasodilation

Leprosy is a granulomatous disease primarily affecting the peripheral nerves. The pathogenesis would be related to the cell-mediated response to mycobacterial antigens, metabolic and biochemical change of Schwann cell, circulating demyelinating factors and other autoimmune process. A specific nerve tissue protein, S-100 protein, has been demonstrated in normal nerves and nerve complexes. The stains of S-100 protein in dermal nerves of leprosy patients have been suggested in assessing the presence of nerve damage. We have estimated the concentration of S-100 protein in the sera of 64 leprosy patients(38 lepromatous leprosy, 26 tuberculoid leprosy) and that of 20 normal controls without neurologic disorders by ELISA. The results obtained were as follows: 1. The mean S-100 protein concentration was 0.0042ng/ml in a total of 64 leprosy patients, with 0.0062ng/ml in lepromatous type and 0.018ng/ml in tuberculoid type. The controls showed 0.0017ng/ml. 2. The analysis of age and serum S-100 protein concentration in both types showed lower value in the fifties of tuberculoid type(p<0.05). With the increase of age, mean S-100 protein concentration in both types tended to increase, but there was no significant correlation(p>0.05). 3. The analysis of duration of illness and serum S-100 protein concentration in both types showed higher value in the forties and fifties in lepromatous type(p<0.05). With the increase of duration of illness, mean S-100 protein concentration tended to increase in lepromatous type and slightly decreased in tuberculoid type, but there was no significant correlation(p>0.05). 4. The mean S-100 protein concentration of patients with neurologic symptoms was 0.0577ng/ml, in contrast with 0.0016ng/ml in patients without neurologic symptoms (p<0.05). In conclusion, the measurement of serum S-100 protein would play a potential role of a useful marker of assessing nerve damage in leprosy patients, esp, with neurologic symptoms.
Coloring Agents ; Enzyme-Linked Immunosorbent Assay ; Humans ; Leprosy* ; Leprosy, Lepromatous ; Nerve Tissue ; Nervous System Diseases ; Neurologic Manifestations ; Peripheral Nerves ; S100 Proteins*

Solar urticaria is an uncommon photodermatosis characterized by the occurrence of pruritic erythema and wheals after exposure to sunlight or artificial radiation. A 52-year-old woman presented with a month long history of pain, wheal and sometimes mild dyspnea which developed after sunlight exposure. Phototesting for UVA, UVB and visible light were all positive, which has never been reported in Korea. The patient was treated with antihistamine and antimalarial drug.
Dyspnea ; Erythema ; Female ; Humans ; Korea ; Light* ; Middle Aged ; Sunlight ; Urticaria*

Celecoxib (Celebrex(R)) is a nonsteroidal anti-inflammatory drug (NSAID) which inhibits cycloxygenase (COX)-2 selectively. It is widely used in osteoarthritis and rheumatoid arthritis patients because it causes less gastrointestinal injury than conventional NSAIDs. We report a case of erythema multiforme probably induced by this drug. A 63 year old female patient had taken celecoxib for osteoarthritis and 8 days after she developed erythema multiforme on the whole body.
Anti-Inflammatory Agents, Non-Steroidal ; Arthritis, Rheumatoid ; Drug Eruptions ; Erythema Multiforme* ; Erythema* ; Female ; Humans ; Middle Aged ; Osteoarthritis ; Celecoxib

BACKGROUND: The pathophysiological and neurochemical changes following spinal injury are not yet elucidated. This study was designed to evaluate the morphological changes of the dorsal horn of the spinal cord and profiles of pain behaviors following intraspinal injection of NMDA in rats. METHODS: Rats were randomized into three groups: a sham-operated control group and groups where the rats received 10 mM or 100 mM N-methyl-D-aspatate (NMDA) injected into their spinal dorsal horn. Following injection, hypersensitivity to cold and mechanical stimuli and excessive grooming behaviors were assessed serially for four weeks. Morphological changes of the spinal cord were evaluated four weeks after intraspinal injection. RESULTS: Few animals in the NMDA groups developed hypersensitivity to cold and mechanical stimuli. The number of groomers and the severity of excessive grooming were significantly higher in the 100 mM NMDA group than those values of the control and 10 mM NMDA groups. The size of the neck region (lamina III-IV) was significantly smaller in the 100 mM NMDA group than in the control and 10 mM NMDA groups. CONCLUSIONS: In conclusion, intraspinal injection of NMDA in rats leads to the pathological sequela in the spinal cord and to excessive grooming behavior. These results support the use of NMDA and excessive grooming behavior after excitotoxic SCI as a model to study chronic pain after SCI.
Animals ; Chronic Pain ; Cold Temperature ; Grooming ; Horns ; Hypersensitivity ; Injections, Spinal ; N-Methylaspartate ; Neck ; Pilot Projects ; Rats ; Spinal Cord ; Spinal Cord Injuries ; Spinal Injuries