Epstein-Barr virus (EBV) is a potent inducer of polyclonal B lymphocyte proliferation and a tool for the establishment of human B lymphoblastoid cell lines (B-LCLs), which have proven useful for several human immunologic applications. B-LCLs serve as efficient antibody-producing cells and antigen-presenting cells. In spite of these advantages, the cloning efficiency of B-LCLs is less than 1%. In order to generate clones of B-LCLs, we cultured B-LCLs with and without canine stromal cell line, DO64, as feeder cell which was immortalized by transduction of retrovirus encoding E6 and E7 of the human papilloma virus type 16 (HPV-16), which was defined to produce various cytokines including stem cell factor (SCF) and interleukin- 6 (IL-6). After 3 weeks of B-LCLs cultured with DO64, 8.3% and 37.5% in 1 cell and 3 cells per well were efficiently cloned, respectively. There was no significant effect on growing of 8-LCLs without DO64 cells and on high concentration of FBS. The cloning efficiency of B-LCLs transduced by retrovirus cultured with and without DO64 cells was 4.2% and 0% in 3 cells per well, respectively, while that of stable transfectant 33.3% and 8.3% in 1 cell per well, respectively. Our results suggest that the use of DO64 cells as feeder cells might permit the cloning of B-LCLs. This efficient cloning of B-LCLs could be used for the convenient source of autologous antigen-presenting cells expressing foreign antigen for the study of human immune responses in vitro, and for a variety of additional purposes, such as the production of human monoclonal antibodies.
Antibodies, Monoclonal ; Antibody-Producing Cells ; Antigen-Presenting Cells ; Cell Line* ; Clone Cells* ; Cloning, Organism* ; Cytokines ; Feeder Cells* ; Herpesvirus 4, Human ; Humans* ; Lymphocytes ; Papilloma ; Retroviridae ; Stem Cell Factor ; Stromal Cells*

We define acute respiratory failure(ARF) as present whenever the ratio of arterial oxygen tension (PaO2) and inspired oxygen concentrarion(FiO2) is below the normal predicted range for the patient's age and/or the arterial carbon dioxide tension(PaCO2) is above 45 in the absence of respiratory compention for metabolic alkalosis. Adult respiratory distress syndrome(ARDS) which is associated with shock, trauma, infection, inhalation of toxic gas, aspiration of gastric content and drugs etc, first received wide-spread attention in 1967. ARDS is a descriptive term that is characterized by a combination of refractory hypoxemia and severly decreased lung compliance. Numerous specific incidents or illnesses may be complicated by, or associated with ARDS. Early diagnosis and improvement in physiological therapy including PEEP therapy have been successful in treating the early and mild episodes of ARDS. This report describes three cases of ARF following anesthesia and reviews the literature.
Adult ; Alkalosis ; Anesthesia* ; Anoxia ; Carbon Dioxide ; Early Diagnosis ; Humans ; Inhalation ; Lung Compliance ; Oxygen ; Respiratory Insufficiency* ; Shock

The effects of nifedipine, a dihydropyridine Ca2+ antagonist, on the eleetrically-evoked twitch response, train-of-four and tetanic stimulation were studied in the isolated rat hemidiaphragm preparation. Nifedipine, in concentrations ranging from 3 to 100 uM, increased the electrically-evoked (nerve stimulation, 0.1 Hz, 0.5 ms, 10 V) twitch response and train-of-four ratio in a dose-relat- ed fashion, and the potentiating effects were inhibited by d-tubocurarine preteratment. The effect of nifedipine was not affected by reducing the extracellular Ca2+ concentration from 2.5 mM to 1.25 mM. In cases of the direct(muscle, 0.1 Hz, 5 ms, 10 V) stimulation, nifedipine increased the twitch response in a dose-dependent manner, but the amplitudes were smaller than those in indirect stimulation. Nifedipine 30 uM potentiated the contractile response induced by 70 mM KC1 and caffeine(10 mM)-induced isometric contractile responses were markedly potentiated by nifedipine treatmeat. Nifedipine 70 upotentiated the effect of l mM caffeine on the electrically-evoked twitch response and the potentiating effect was also seen in reverse treatment. On the basis of these findings, the result of present study suggests that the potentiating contractile response by nifedipine is mediated by two distinctive mechanisms. One is the acetylcholine release from presynaptic nerve terminal and the other may be due to the releases of Ca2+ in sarcoplasmic reticulum.
Acetylcholine ; Animals ; Caffeine ; Nifedipine* ; Rats* ; Sarcoplasmic Reticulum ; Tubocurarine

In an attempt to evaluate the cardiotoxicity of bupivacaine, beating rate, tetrazolium MTT and lactate dehydrogenase activity were investigated in the medium containing bupivacaine for 24 hours after neonatal rat myocardial cells were cultured for 72 hours. Light and electron microscopic studies were also carried out. The results were as follows ; 1) Beating rate decreased dose-dependently, and beating cells were not observed over 10(-4) M concentration of bupivacaine. 2) MTT50 value was 0.32 ug/ml (1,000 uM). 3) The amount of lactate dehydrogenase released into the medium was 192% of control cells at 10(-3) M concentration of bupivacaine. 4. In light microscopy, myocardial cells were decreased in number dose-dependently, and showed a few cytoplasmic processes and lots of granules in cytoplasm at 10 M concentration of bupivacaine. 5. Electron microscopy of bupivacaine-treated cells showed smooth endoplasmic reticulum, destruction of mitochondria and Golgi apparatus and increase of vacuoles and dense bodies. It also showed dilatation of rough endoplasmic reticulum and loss of myofibrils. These results suggest that high concentration of bupivacaine (> or = 10(-4) M) induee remarkable toxicity on cultured rat myocardial cells.
Animals ; Bupivacaine* ; Cytoplasm ; Dilatation ; Endoplasmic Reticulum, Rough ; Endoplasmic Reticulum, Smooth ; Golgi Apparatus ; L-Lactate Dehydrogenase ; Microscopy ; Microscopy, Electron ; Mitochondria ; Myofibrils ; Rats* ; Vacuoles

Low molecular weight dextran (dextran 40) has been widely used not only as a plasma expander and blood flow improver in various diseased or shock states, but also to prevent postoperative thromboembolic complications, especially in major operations such as vascular and hip surgery. Despite the fact that the drug can be administered with few complications, it can produce serious lifethreatening anaphylactoid reactions, although this occurs very rarely. The authors retrospectively investigated 89 patients, who developed a shock state perioperatively, out of 5267 patients in the operating room from April 1975 to June 1980 at the Jeonbug National University Hospital. Eight patients out of 89 developed a shock state with flushed skin, goose skin, bronchospasm, conjunctival injection and chest pain. These symptoms and signs were thought to be due to an anaphylactoid reaction to dextran 40. During this period of time, 2023 dextran 40 units were administered, so that the incidence of dextran induced anaphylactoid reaction was up to 0.40%. This is more than 10 times as much as reported by some investigators.
Bronchial Spasm ; Chest Pain ; Dextrans* ; Hip ; Humans ; Incidence ; Jeollabuk-do ; Molecular Weight ; Operating Rooms ; Plasma ; Research Personnel ; Retrospective Studies ; Shock ; Skin

Arnold-Chiari malformation type I (ACM I) is anatomically defined as the displacement of the cerebellar tonsils below the level of the foramen magnum. Syringomyelia is a condition in which a cavity called a syrinx develops in the spinal cord and is filled with cerebrospinal fluid. Here we report the anesthetic management of a case of ACM I with associated syringomyelia scheduled for suboccipital craniectomy, cervical laminectomy and duraplasty.
Arnold-Chiari Malformation ; Displacement (Psychology) ; Foramen Magnum ; Humans ; Laminectomy ; Palatine Tonsil ; Spinal Cord ; Syringomyelia

BACKGREOUND: The sympathetic investigations during thoracic sympathectomy are essential to an adequate sympathectomy that will lead to sufficient and lasting relief of palmar hyperhidrosis. The measurement of palmar skin temperature has been used as an indicator of success of transcutaneous chemical thoracic sympathectomy. We measured intraoperative palmar skin temperature to know whether it can be used as a same purpose in the endoscopic thoracic sympathectomy under general anesthsia. METHODS: Fifteen patients (18 to 25 years old) with palmar hyperhidrosis underwent endoscopic thoracic sympathectomy under general anesthesia. The palmar skin temperature was measured with a skin probe of a thermometer applied on the both index finger tips. The palmar skin temperature was monitored continuously from the beginning of anesthesia to the complete arousal. RESULTS: The palmar skin temperature increased significantly by about 3 degrees C just after induction. There was no significant difference in the palmar skin temperature between just before sympathectomy and soon after sympathectomy during the endoscopic thoracic sympathectomy. CONCLUSIONS: Intraoperative measurement of palmar skin temperature can not indicate a definite sympathectic denervation during the endoscopic thoracic sympathectomy under general anesthesia.
Anesthesia ; Anesthesia, General ; Arousal ; Denervation ; Fingers ; Humans ; Hyperhidrosis* ; Skin Temperature* ; Skin* ; Sympathectomy* ; Thermometers

Many techniques have been tried to avoid the adverse effect of succinylcholine administe-red for endotracheal intubation especially wish the complication of increased IOP, hyperk-alemia, aspirationl pneumonia and post operative muscle pain, One of these is that the prior administration of a small, subparalyzing dose(15 ug/kg) of non-depolarizing muscle relaxant would shorten the onset time of an intubating dose(80 ug/kg) of muscle relaxant. Intra-venous lidocaine has bean effective in attenuating the reflex intra-ocular response to laryngoscopy Therefore, we determined the effectiveness of this drug regimen with and without intra venous lidocaine to attenuate the IOP, blood pressure and heart rate response to laryngos-copy and endotracheal intubation. Forty patients were divided intro two groups. Group l (n=20) administered saline 5 ml. Group ll(n=70) administered 2% preservative free lidocaine(1.5 mg/kg) as pretreatment drug. The results are as follows : 1) There was no statistically significant difference of intubation condition between one and another group. Among the forer patients, Grade 1,2,3,4 are 8(20%), 19(47.5%), 13(32.5%),0, in orders. 2) In the Saline Group, IOP, BP, HR increased significantly after laryngoscopy compared wiith control value. (p<0.001, P<0.05/p<0,001, p<0.005) and maintained above control values to 4~5 minutes later. 3) In the Lidocaine Group, IOP, BP, HR increased slightly after laryngoscopy compared with control value, but thege changes were not statistically significant, and decreased below control values in 2 min, 2min, 3 min after laryngoscopy each to each. From the above results, it is suggested that combined method of pretreatment of 2% lidofaine(1.5 mg/tg) and divided dose of pancuronium is valuable in general anesthesia of ophthalmic patient who need to attenuate the IOP.
Anesthesia, General ; Blood Pressure ; Heart Rate ; Humans ; Intubation ; Intubation, Intratracheal ; Laryngoscopy ; Lidocaine ; Myalgia ; Pancuronium* ; Pneumonia ; Reflex ; Succinylcholine

Atropine and glycopyrrolate have been used for the reversal of non-depolarizing neuromuscular blockade as anticholinergics. Atropine may produce the central anticholinergic effect because it is a tertiary amine which can cross the blood-brain barrier. Glycopyrrolate is a quaternary ammonium compound that does not cross the blood-brain barrier. and as a result, it has no significant central anticholinergic effect. This report compares postanesthetic arousal between glycopyrrolate-neostigmine and atropine-neostigmine mixtures. We conclude that the glycopyrrolate-neostigmine mixture group have significantly more rapid arousal than the atropine-neostigmine group.
Ammonium Compounds ; Arousal* ; Atropine ; Blood-Brain Barrier ; Cholinergic Antagonists ; Glycopyrrolate ; Neuromuscular Blockade

The effects of diltiazem and verapamil on the electrically-evoked twitch response, train-of- four and tetanic stimulation were studied in the isolated rat hemidiaphragm preparation. Diltiazem(3-150 pM) and verapamil(3-100 pM) increased the electrically-evoked(nerve stimulation, 0.1 Hz, 0.5 ms, 10 V) twitch responses in a dose-related fashion and diltiazem was more potent than verapamil. But, the large doses of diltiazem(150-300 uM) and verapamil(100-300 uM) decreased the twich responses. And the effects of diltiazem and verapamil were not effected by reducing the extracellular calcium from 2.5 to 1.25 mM. Diltiazem and verapamil decreased the train-of-four and tetanus ratio as well as the d-tubocurarine in a dose-related fashion. d-Tubocurarine, a specific nicotinic antagonist, decreased twitch response, and the potentiating twitch response of diltiazem was significantly inhibited by pretreatment of d-tubocura- rine. Furthermore, it is noteworth that the inhibitory effects of d-tubocurararine were markedly potentiated by diltiazem. In cases of the direct(muscle, 0.1 Hz, 5 ms, 10 V) stimulation, diltiazem and verapamil decreaaed the electrically-evoked twitch response with dose dependently. These results indicate that diltiazem and verapamil elicited two distinctive types of twitch response in the rat phrenic-hemidiaphragm preparation. The potentiating effect of twitch response is mediated by the acetylcholine release from the prejunctional nerve terminal and the inhibiting effect may be due to blcking influx of calcium and/or release of acetylcholine from presynaptic nerve terminals.
Acetylcholine ; Animals ; Calcium ; Diltiazem* ; Rats* ; Tetanus ; Tubocurarine ; Verapamil*