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The effect of various non-depolarizing neuromuscular blocking agents (gallamine, pancuronium, vecuronium, d-tubocurarine, metocurine, atracurium and pipecuronium) on [³H] acetylcholine release in the response to field electrical stimulation was investigated in vitro in preparations of the guinea pig right atrium. In this preparation, atropine enhanced and oxotremorine, a muscarinic agonist, reduced the release of [³H] acetylcholine. Atropine reversed the inhibitory effect of oxotremorine in a concentration dependent manner, indicating that there is negative feedback modulation of acetylcholine release from the vagal nerve. While pancuronium, gallamine and atracurium enhanced the release of [³H] acetylcholine, d-tubocurarine, metocurine, vecuronium and pipecuronium did not affect it. Pancuronium and gallamine also reduced the inhibitory effect of oxotremorine and the K_d value of pancuronium for muscarinic receptors located on cholinergic nerve terminals was 2.31 μM. These findings indicate that pancuronium and gallamine enhanced the release of acetylcholine from the atrial parasympathetic nerve, probably by inhibiting presynaptic muscarinic receptors.

We report a method of endoscopic retrograde sphincterotomy in patients in whom the optimal viewing of the papilla of Vater is hardly obtained. Percutaneous transhepatic cholangiodrainage (PTCD)-tube is placed under ultrasonographic guidance. PTCD-tube coming out of the papilla of Vater is observed by the endoscope and the guide wire inside the shpincterotome is inserted into the PTCD-tube. Sphincterotome is advanced into the common bile duct by the guidance of the guide wire and PTCD-tube. Sphincterotomy is performed in a usual fashion. Two patients with previous history of gastrectomy underwent this procedure with successful results. This method should be tried when usual method of EST is difficult and unsuccessful.

The axial length of the eye, the radius of the anterior corneal surface, and the diameter of the cornea were measured on 40 eyes of 20 New Zealand White rabbits. The ultrasonographic measurements of the axial length of the eye showed a mean value of 15.12 ± 0.51mm (15.33 ± 0.50mm in males, 14.96 ± 0.46mm in females). The mean corneal radius was 7.26 ± 0.26mm (7.18 ± 0.9mm in males, 7.32 ± 0.33 mm in females); the mean value of the difference in corneal curvature between two principle meridians was 0.21 ± 0.12mm; and the mean horizontal and vertical diameters of the cornea were 13.41 ± 0.34mm and 13.02 ± 0.30mm, respectively. Although the diameter of the cornea of rabbits was bigger than that of humans (average, horizontal 11.75mm, vertical 10.55mm), the axial length of the eye was 0.7 times that of humans (23-24mm), and the radius of corneal curvature was relatively smaller than that in humans (7.7-7.8mm).

We studied deformity of the proximal end of the femur following open reduction using the wide exposure method for developmental dislocation of the hip. We reviewed radiographs of 22 children with unilateral dislocation of the hip who had undergone open reduction between one and three years of age. Ages at final examination ranged from 14 to 21 years. None of the patients in this study had undergone any additional surgery. Avascular necrosis of the femoral head was not observed in any patients. Radiographic measurements were observed when the children were 3, 6, 9, and 12 years old, and at the final examination. Coxa magna and valga were the factors associated with poor results. Coxa valga had occurred at 6 to 12 years of age, but rapidly improved thereafter. Throughout the course of treatment the epiphysis-femoral neck angle was within the normal range. The fusion of the epiphyseal growth plates on both sides occurred simultaneously. The coxa valga was due to valgus of the femoral neck, and not to valgus head tilt. These results suggest that, provided the mechanics of the joint have been properly corrected as in our operation, the biomechanics of the hip will improve until growth ceases, but that it may take more than ten years to attain normality.

 We studied the clinical efficacy of roxithromycin (RXM) administered at the daily dosage of one tablet (150mg) for 3 months in 30 patients with chronic sinusitis. The effectiveness of this drug was evaluated on a four-point scale. Subjective and objective symptoms disappeared or decreased markedly, especially postnasal drip and nature of discharge in 80 percent or more of the patients. All symptoms significantly decreased (P < 0.001; headache P < 0.05), except for the sensation of foul odor. Symptoms improved even in those cases in which Haemophilus influenzae was detected. It is suggested that RXM produces some clinically beneficial effect through an immunological and or anti-inflammatory mechanisms in addition to its antibiotic effect.

Malignant lymphoma was induced in Japanese (JWY), New Zealand (NZY) and Dutch (DUY) white rabbits by oral spray of cell-free pellets of culture fluid (crude virus fraction) of Ts-B6 cells (cynomolgus monkey B-lymphoblastoid cells harboring Epstein Barr virus-related simian herpesvirus or Cyno-EBV). Nine of 11 inoculated rabbits developed malignant lymphomas within 42-160 days after oral inoculation (JWY, 2/3; NZY, 5/6; DUY, 2/2). In contrast, none of the control rabbits inoculated in the same fashion with B95-8 (EBV-producing marmoset cell line) cell-free pellets developed malignant lymphoma. Most rabbits showed increased anti-VCA IgG and anti-EA-DR IgG antibody titers after inoculation by oral spray of Ts-B6 cell-free pellets. EBV-encoded RNA-1 was revealed in the tumor cells by in situ hybridization. EBV DNA was detected in the rabbit peripheral blood leukocytes (PBL) by polymerase chain reaction; the earliest positive result was obtained only two days after oral inoculation. These data suggest that orally administered Cyno-EBV in Ts-B6 cells infects PBL and then induces malignant lymphoma in rabbits. The availability of this animal model promises to clarify the role of EBV in human lymphoma and provides a means for studying prophylactic and therapeutic regimens.

A retrospective study on postoperative complications and factors affecting prognosis was performed on elderly patients with gastric cancer. We studied the correlation of age, pathological depth, preoperative laboratory data, physical status, duration of surgery, volume of blood loss, blood transfusion, curability, and extent of lymph node dissection to postoperative complications and prognosis in 47 patients with gastric cancer over 80 years old. Preoperative function of lung and liver frequently showed abnormal data. Postoperative complications were noted in 47% of patients, especially in the pulmonary system, liver and heart. Curability and extent of lymph node dissection were the significant factor affecting survival. Some mortalities caused by initial malignancy were recognized in the conservative lymph node dissection in the stage I . The incidence of postoperative complications was not significantly different according to extent of lymph node dissection. Blood transfusion was the only significant factor for the incidence of postoperative complication. The most frequent cause of death was the initial malignancy. We recommend that a low grade lymph node dissection should not be readily chosen for elderly patients in early cases.

 The aim of the study was to investigate whether Na⁺ channels play a role in the twitch component of the response of the isolated frog rectus abdominis to Ca²⁺-free Ringer solution with 0.2 mM Na₂EDTA by using tetrodotoxin and some other well known drugs that exhibit a blocking action on Na⁺ channels. In the presence of 5 x 10^-7 M tetrodotoxin, the twitch component, measured isotonically, disappeared. Although 10^-7 M d-tubocurarine was found to be ineffective, a complete blockage of twitch amplitude was observed at 5 x 10^-6 M concentration of the drug. The inhibitory action of d-tubocurarine on twitch response was not antagonized by 10^-6 and 10^-5 M carbachol. Propranolol (10^-6 - 10^-5 M), lidocaine (2 x 10^-6 - 10^-5 M), quinine (10^-6 - 2 x 10^-5 M) and quinidine (10^-6 - 2 x 10^-5 M) inhibited maximal twitch amplitude in a concentration dependent manner. These findings strongly suggest that activation of tetrodotoxin sensitive Na⁺ channel may play a primary role at twitch generation during exposure of the frog rectus abdominis to Ca²⁺-free Ringer solution with Na₂ EDTA.

Physiological strain plays an important role in maintaining the normal function and metabolism of bone cells. It is well known that the mineral content of astronauts' bones decreases during spaceflight. Thus, gravity is one of the important factors in the muscloskeletal system. The vector-free horizontal clinostat has been used to simulate conditions of microgravity for examining such effects on cells in culture. We analyzed the effects of simulated microgravity using a horizontal clinostat on cultured osteoblast-like cells (HuO9 cell line). Total cellular protein, which was measured as an indication of cell proliferation, was not significantly inhibited under simulated microgravity conditions. No morphological changes were detected under microgravity conditions by phase-contrast microscopy. However, the alkaline phosphatase (ALP) activity and osteocalcin production of the HuO9 cells decreased significantly under microgravity conditions. Our data indicate that simulated microgravity directly inhibits some differentiation phenotypes and some functions of osteoblasts. On the other hand, the addition of 1,25-dihydroxy-vitamin D₃ (1,25-(OH)₂-D₃) increased ALP activity under simulated microgravity conditions, although the total activity of ALP in the cells treated with 1,25-(OH)₂-D₃ was still lower under simulated microgravity conditions than that in the control cells. However, the cells under simulated microgravity conditions showed a greater enhancement of ALP activity by treatment with 1,25-(OH)₂-D₃.

Our aim was to study the effect of iron supplementation on the following aspects of erythrocyte metabolism in experimental hyperthyroidism: glutathione (GSH) levels, glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) activities. Hyperthyroidism induced by L-thyroxine administrations significantly raised erythrocyte GSH, GSH-Px and SOD levels of the rats (P < 0.001). Likewise, we observed that iron supplementation induced significant rises in erythrocyte GSH, GSH-Px and SOD levels (P < 0.001) as compared with the control group. The erythrocyte GSH, GSH-Px and SOD levels of hyperthyroidism-induced iron-supplemented animals were significantly higher when compared with either the iron-supplemented group (P < 0.001) or the only L-thyroxine-administered hyperthyroid group (P < 0.001, P < 0.05, P < 0.01, respectively). The results of this study show that L-thyroxine administration and/or iron supplementation increases GSH, GSH-Px and SOD levels of erythrocytes.

We studied 92 patients with allergic rhinitis in Syodoshima, Japan, during the pollen season between April and June to evaluate the cross-reactivity to different antigens, including pollen from the olive tree (Olea europaea) and orchard grass (Dactylis glomerata). Olive tree pollen was present in the atmosphere for 23 days, from May 19 to June 12, 1994. Specific IgE antibodies for olive tree pollen antigen were present in 21 (26.9%) of the 78 patients with allergic rhinitis. Nine (24.3%) of the 37 patients with allergic rhinitis exhibited positive skin reactivity to an extract of olive tree pollen. Fifteen (88.2 %) of the 17 patients who had IgE reactivity in their sera to olive tree pollen antigen demonstrated allergic reactions to an extract of olive tree pollen. Specific IgE antibodies for orchard grass pollen antigen were present in 43 (48.3%) of the 89 patients with allergic rhinitis and 20 (95.2%) of the 21 patients who had IgE reactivity in their sera to olive tree pollen antigen. The inhibition test using the CAP System revealed that the reactivity of the IgE antibody specific for olive tree pollen antigen was inhibited dose-dependently by an extract of orchard grass pollen. These findings show that there is a reaction in some patients with grass (Gramineae) pollinosis that might be induced by olive tree pollen.

We have established an Adriamycin (ADM) -resistant small cell lung cancer (SCLC) cell line, SBC-3/ADM100, which shows multifactorial mechanisms of resistance to ADM, such as overexpression of P-glycoprotein, an enhanced detoxifying system and a decrease in topoisomerase II activity. In the present study, we confirmed that SBC-3/ADM 100 showed collateral sensitivity to methotrexate and TNP-351, a new antifolate, though this cell line showed a typical multidrug resistance (MDR) pattern. We also demonstrated a faster uptake and higher accumulation (1.3-fold) of TNP-351 in the SBC-3/ADM100 cells than those in the parent SBC-3 cells. These results explain one of the mechanisms for collateral sensitivity in the resistant cells. Furthermore, this cell line was found to have no cross-resistance to edatrexate and minimal cross-resistance to trimetrexate, 254-S (cisplatin analog), 5-fluorouracil and 4-hydroperoxyifosfamide. These drugs will have clinical importance in patients with SCLC who were previously treated with an ADM-containing regimen. Thus, antifolates, especially TNP-351 and edatrexate, can be expected to eradicate residual multidrug resistant SCLC cells selected by ADM.

Sixty-seven trigger fingers of 58 idiopathic and hemodialysis patients were treated by percutaneous A1-pulley release technique. Severity of triggering was classified into five grades for treatment selection and prediction of possible results. Results were excellent in 41 fingers, good in 9, fair in 7, and poor in 10, requiring additional treatment. The results of the lower grades were better, and those of the higher grades were poor. Excellent or good results appeared to depend on the proper selection of the patients according to the grading system and confirmation of triggering disappearance just after the release. There were neither infections nor neuro-vascular deficits after treatment. Compared to conventional open release, this treatment was found to be more useful from the standpoints of ease and safety of the technique, and the patients' quick return to normal life.

We compared the transfection efficiency of four types of positively charged liposomes composed of (i) N-(α-trimethylammonioacetyl)-didodecyl-D-glutamate chloride (TMAG), dilauroylphosphatidylcholine (DLPC), and dioleoylphosphatidylethanolamine (DOPE) (1:2:2 molar ratio); (ii) 3β [N-(N′, N′-dimethylaminoethane)-carbamoyl] cholesterol (DC-Chol) and DOPE (3:2 molar ratio); (iii) dimethyldioctadecylammonium bromide (DDAB) and DOPE (1:2.2 molar ratio); (iv) N-[1-(2,3-dioleyloxy) propyl] -N,N,N-trimethylammonium chloride (DOTMA) and DOPE (1:1, w/w; lipofectin). Luciferase gene was used as a reporter gene. Among the cationic liposomes used, the liposomes composed of TMAG, DOPE and DLPC showed a much higher efficiency of plasmid DNA entrapment than the other cationic liposomes tested. In the absence of serum, the cationic multilamellar vesicles (MLV) and small unilamellar vesicles (SUV) composed of TMAG, DOPE and DLPC gave highly efficient transfection. On the other hand, MLV, dehydration-rehydration vesicles (DRV), and SUV liposomes prepared with the mixtures of DC-Chol and DOPE showed similar levels of transfection efficiency. However, the cationic liposomes composed of DDAB and DOPE showed inferior efficiency, whether in the form of DRV, SUV or MLV. The transfection efficiency of lipofectin was also low. In the presence of serum, on the other hand, a considerable (about 30-50%) amount of transfection activity was still observed at 10% fetal calf serum in the cationic MLV and SUV composed of TMAG, DOPE and DLPC. Cationic MLV, composed of TMAG, DOPE and DLPC, can transfect plasmid DNA, not only in the adherent cell lines but also in the suspension cell lines. These findings indicate that the transfection efficiency of cationic liposomes is affected by the lipid composition, the type of liposome, or the presence or absence of serum. They also indicate that the cationic liposomes containing TMAG, DOPE and DLPC are efficient vectors for gene transfer into cells.

Accurate assessment of elbow function is important to determine the total ability of the arm. The purpose of this study was to clarify the relationship between isometric muscle strength of the elbows of patients with rheumatoid arthritis (RA) and Larsen's X-ray evaluation. Fifty-six elbows of 45 RA patients aged 47 to 77 years (mean age, 63 years) were tested. Muscle strength was measured with an isometric torque-cell dynamometer. Test-retest reliability of the dynamometer was proven by measuring 12 elbows of 6 healthy young men. In RA patients, elbow flexion and extension strength decreased in proportion to increases in the severity of Larsen's grades from Grade 1 to 4. However, Grade 5 elbows had greater muscle strength than those in Grade 4. Forearm pronation and supination strength also decreased in proportion to increases in the severity of Larsen's grades from Grade 1 to 5. This quantitative study made it clear that the muscle strength of RA patients' elbows almost completely correlates to X-ray finding according to the grade of Larsen's evaluation based on X-rays. With regard to muscle strength of postoperative elbows, both flexion strength and supination strength after total elbow replacement (TER) were about two times greater than before TER, and after synovectomy it was as great as those in non-operative RA patients of Grade 2.

A case of a male patient with Kniest dysplasia is reported. The patient's growth and the development and evolution of the patient's abnormalities were tracked for a 30 year period, starting at the patient's birth. The clinical and radiographic features during this period, along with the differential diagnosis of Kniest dysplasia, are discussed. Femoral capital epiphyses and the presence of a cataract in one eye were noted from the early stages of the patient's life. The patient's final height was 165 cm. We believe this to be the first long-term follow up of this condition.

The expression of osteonectin (ON) in osteoarthritic articular cartilage was investigated by enzyme immunohistochemistry and colloidal gold immunoelectron microscopy. A total of 96 specimens from 9 knees of 8 patients with osteoarthritis (OA) were examined. In OA cartilage, ON-positive cells varied in distribution and were not seen in all the specimens obtained from the same patient. However, in over half of the specimens (56 of 96), especially in the specimens of Mankin's grades from 4 to 9, which corresponds to relatively early stages of OA, ON was expressed in the cartilage above the calcified layer. On the other hand, ON was detected only in the calcified layer below the tidemark in normal articular cartilage. In addition, colloidal gold immunoelectron microscopy revealed ON in chondrocytes and matrix vesicles (MVs). These findings suggest that ON acts through MVs in the early stages of OA as a significant pathogenetic factor involved in intracartilage calcification, which is known to have a close relationship to the progression of OA.

We used a yeast functional assay (functional analysis of separated alleles in yeast: FASAY) to determine the p53 gene status of human cell lines maintained in our laboratory. This assay enables the researcher to score wild-type p53 expression on the basis of the ability of expressed p53 to transactivate the reporter gene HIS 3 via the p53-responsive GAL 1 promoter in Saccharomyces cerevisiae. The cell lines examined were ten hepatoma, two hepatoblastoma, three in vitro immortalized fibroblast, two osteosarcoma, a chondrosarcoma, an ovarian teratocarcinoma and a colon cancer cell line. Out of 20 cell lines, 11 cell lines had mutations in both alleles of the p53 gene, and another 8 cell lines had no mutation in the p53 gene. Thus, 55% of the cell lines examined had mutations in the p53. Interestingly, PA-1 cells had both the normal and the mutant p53 alleles, showing that FASAY is a useful method for detecting the wild-type and mutated p53 genes simultaneously. As for the three liver cell lines harboring HBsAg, there was no relationship between their p53 gene status and the presence of HBsAg. Two cell lines were normal for p53 status, while the other had a mutation of the p53 gene.

Retinal cells from chick embryos aged 7.5 days of gestation were cultured for two months in a non-adherent suspension culture dish to study the effects of growth factors and co-culture with retinal pigment epithelial cells on their differentiation. Dissociated retinal cells became cellular aggregates (multicellular spheroids) within a day, and rosettes were formed in the spheroids after 2 days. Ultrastructurally, neurons of the rosettes developed connecting cilia, ellipsoids (accumulation of mitochondria), and external limiting membrane, indicative of their differentiation into photoreceptor cells. Epidermal growth factor enhanced the expression of rhodopsin by rosette-forming neurons, while basic fibroblast growth factor induced the growth of Mueller cells at 4 weeks, and their transdifferentiation into lens-epithelial-like cells at 8 weeks. Co-culture of retinal cells with retinal pigment epithelial cells enhanced the formation of rosettes in spheroids. Multicellular spheroids formed in a dish for suspension culture would provide a convenient in vitro system to examine differentiation and transdifferentiation of the retina.

UDP-galactosyltransferase (UDP-Gal-T) is a key enzyme in the synthesis of mucus glycoprotein which plays an important role in gastric mucosal defensive mechanisms. Analysis of gastric UDP-Gal-T activity should clarify the mechanisms of the action of antiulcer drugs regarding gastric defensive factors. Here, we examined UDP-Gal-T activity in rat gastric mucosa treated with the antiulcer drugs geranylgeranylacetone (GGA) and cetraxate hydrochloride (CET). The effects of coadministration of indomethacin and exogenous administration of prostaglandins (PGs) were also studied. GGA and CET significantly increased UDP-Gal-T activity, and coadministration of indomethacin inhibited the increase of enzyme activity. UDP-Gal-T activity level with GGA was significantly higher than the control level, even in the presence of indomethacin. With CET, however, this was not the case. Among PGs, PGE1 significantly increased enzyme activity. Concomitant administration of PGE1 and GGA or CET increased UDP-Gal-T activity even with indomethacin to the levels achieved when these antiulcer drugs were administered without indomethacin. Our findings suggest that GGA and CET exert antiulcer effects by increasing mucus glycoprotein synthesis and that endogenous PG synthesis may be involved in this process. However, mechanisms not mediated by endogenous PGs may also exist in the stimulatory action of GGA on UDP-Gal-T activity.