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*

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

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

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

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

We had a 52-year-old male presented for a surgery for old fracture of the 7th cervical vertebra. Generalized muscle rigidity and high fever with tachycardia was revealed 25 minutes after succinylcholine and enflurane administration. Then the body temperature was risen from 37.8 degrees C to 40.5 degrees C within 20 minutes and the arterial blood gas analysis showed a severe respiratory and metabolic acidosis. Under the suspicion of malignant hyperthermia, all anesthetics were discontinued, refrigerated intravenous solutions were started, and gastric lavage with cold saline was initiated together with packing the body with ice. Oral dantrolene 125mg, the only drug we could obtain, was administered through Levins tube after lavage. Serial serum CPK level were 4414 IU/L on third day and 74 IU/L on seventh day postoperatively. The postoperative course was uneventful and the patient could be walked after fifth day postoperatively and discharged without any sequelae.
Acidosis ; Anesthesia ; Anesthesia, General* ; Anesthetics ; Blood Gas Analysis ; Body Temperature ; Dantrolene ; Enflurane ; Fever ; Gastric Lavage ; Humans ; Ice ; Male ; Malignant Hyperthermia ; Middle Aged ; Muscle Rigidity ; Pharmacology ; Spine ; Succinylcholine ; Tachycardia ; Therapeutic Irrigation

We designed a study to determine if the tracheal tube cuff inflation in the oropharynx improves the success rate of blind nasotracheal intubation in normal, paralyzed patients because of lacking of controlled study about it. In prospective, randomized fashion, 100 ASA I or II patients undergoing elective oral surgery were studied. The trachea was intubated once keeping the tracheal tube cuff deflated throughout the maneuver and once using the technique of tracheal tube cuff inflation in the oropbarynx. A maximum of two attempts was allowed for each technique. If the first attempt was failed, the second attempt was tried with an addition of application of thyroid cartilage compression in each technique. Witb the tracheal tube cuff inflated, the success rate was significantly higher than the cuff-deflated technique(p<0.05). A application of thyroid cartilage compression increased the success rate of the blind nasotracheal intubation in each technique, but it was more useful in the cuff inflation technique(p<0.05). Time taken to intubate the trachea was longer in the cuff inflation technique. We suggest that, in normal paralyzed patients, the tracheal tube cuff inflation in the oropharynx increases the success rate of blind nasotracheal intubation.
Humans ; Inflation, Economic* ; Intubation* ; Oropharynx* ; Prospective Studies ; Surgery, Oral ; Thyroid Cartilage ; Trachea