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An improved direct colorimetric method for determining the concentration of urinary hippuric acid as an index of toluene exposure was described. One tenth ml of urine was diluted with 0.4 ml 0.01 M phosphate buffer H 6.9 and mixed with 0.5 ml pyridine. The mixture was layered on 0.2 ml benzenesulfonyl chloride. The reaction was started by mixing for one min with a mechanical shaker. The colored solution was allowed to stand for 30 min, diluted with 5 ml ethanol, and absorbance measured at 410 nm within 30 min after the dilution. The coefficient of variation of this method was 6% and the recovery 103% when urine contains about 0.2-0.5 mg hippuric acid per ml of urine. The concentration was linear up to 2.0 mg per ml hippuric acid in a specimen.

The chemotactic activity of granulocytes obtained by the Terumo Filtration Leucapheresis System (F.L.) was examined by the method of Boyden's chamber. The number of cells migrating through the Millipore filter was expressed as the chemotactic activity. The mean values were 117 for the F.L. and 122 in a control, in which cells were collected from the same donor blood using dextran sedimentation. The results suggested that the in vitro chemotactic function of granulocytes obtained by F.L. was within normal limits.

To investigate the role of folic acid deficiency in the pathogenesis of anemia in the elderly, hematological examinationa and assays of serum iron, vitamin B12 and folate were carried out on the 86 elderly patients admitted to a home for the aged. Means of red blood cell counts, hemoglobin levels and hematocrit were 385.3 x 10(4)/mm3, 12g/dl and 36%, respectively. These levels were lower than any other report in Japan. Anemia was detected in 23 out of 86 patients. Judging from mean corposcular volume and mean corposcular hemoglobin, most of them were normocytic and normochromic. Although low serum levels of iron and folate were rather frequently observed, the results on hematological examinations suggest that deficiency of these factors alone is not the cause of the anemia in the elderly patients. Rapid clearance of 5-methyl-tetrahydrofolic acid and increased excretion of formiminoglutamic acid after histidine loading were revealed in some of those who had subnormal serum folate levels. Therefore, supplementation of folic acid is recommended to those who had poor dietary intake.

 To determine whether the predominant infiltration with memory CD4⁺T cells in joints is specific to the local immune and inflammatory response in rheumatoid arthritis (RA), the proportions of CD45RA⁺ or CD45RO⁺ cells in the CD4⁺T cell populations in three different compartments (i.e., peripheral blood, synovial fluid, and synovial tissue) from patients with RA and osteoarthritis (OA) were compared by two-color flow-cytometric analysis. In the CD4⁺T cell population of peripheral blood, the number of CD45RO⁺ cells was relatively higher than CD45RA⁺ cells in both RA and OA patients, but their percentages did not differ from those found in healthy individuals. However, the great majority of CD4⁺T cells present in synovial fluid and synovial tissue were CD45RO-positive and CD45RA-negative in both patient groups; although CD4⁺T cells infiltrating both the disease compartments were markedly greater in RA joints, their mean percentages of CD45RO⁺ cells were not significantly different from those in OA joints. These data indicate that an accumulation of CD45RO⁺ memory CD4+T cells is a generalized phenomenon during local inflammatory responses in both RA and OA joints, and may be due mainly to the propensity of these cells to preferentially transmigrate into the inflamed joint via adhesion molecules as compared with CD45RA⁺ naive CD4⁺T cells.

To determine the extent to which recent advances in biomechanical technology have been implemented and to evaluate these new technologies, 84 unilateral above knee (A/K) amputees and their prostheses were surveyed in the Chugoku-Shikoku district of Japan, especially in regard to the types, sockets and components of A/K prostheses currently in use. Background factors such as age and sex of the A/K amputees and the period after amputation were also surveyed. Of the 84 amputees surveyed, 74 (88.1%) were over 40 years old and 40 (47.6%) were over 60 years old. There were 10 women (11.9%) and 74 men (88.1%). The period after amputation was under 25 years in 58 (69.0%) cases. Regarding the type of A/K prostheses, one-third of the prostheses was of the exoskeletal type and two-thirds were of the endoskeletal type. Although the endoskeletal type is becoming more popular recently, elderly A/K amputees tend to use the exoskeletal type. Thirty-one (36.9%) had plug-fit sockets which are preferable for those who follow the Japanese practice of sitting on the floor, especially for elderly amputees. Thirty-seven (44.0%) had a lock-knee, 27 (73.0%) of which were used by amputees over 60 years old. Seventy-three (86.9%) had a single-axis ankle which is generally considered to be the most stable ankle. Thus, the most common combination of prosthetic components for elderly A/K amputees was the plug-fit socket, lock-knee joint and single-axis ankle.

Our aim was to clarify the effect of a somatostatin analogue (octreotide) on glucose flux in conscious dogs. We monitored the effects with catheters in the portal vein, hepatic vein and femoral artery and Doppler flow probes on the portal vein and hepatic artery before and after oral glucose administration. A significant increase of portal vein plasma flow after oral glucose was completely suppressed by both 4 and 1 μg/kg octreotide. All doses of octreotide (4, 1 and 0.1 μg/kg) suppressed the glucose-induced increment of arterial glucose by dose response. Only 4 μg/kg of octreotide slightly but significantly suppressed hepatic glucose output. Marked suppression and delayed glucose absorption by the intestine was observed after 4 μg/kg of octreotide. One and 0.1 μg/kg octreotide also suppressed glucose absorption without delayed absorption. Total amounts of absorbed glucose during 3h after oral glucose were 24 ± 11% with 4 μg/kg of octreotide, 37 ± 16% with 1 μg/kg of octreotide, and 48 ± 8% with 0.1 μg/ kg of octreotide, all of which were significantly less than that of the control (73 ± 8%). Using 4 μg/kg of octreotide treatment, the liver took up only 5 ± 4% of the absorbed glucose, while the liver took up 35 ± 6% and 43 ± 9% of the absorbed glucose with 1 and 0.1 μg/kg of octreotide. These latter values were similar to that of the control value of 34 ± 4%. In conclusion, we found that octreotide administered before oral glucose had a remarkable stabilizing effect on postprandial glycemic surges. Both the direct inhibitory effect of octreotide on portal vein plasma flow and impaired glucose absorption would contribute to this decreased postprandial hyperglycemia, while its suppressive effect on other hormones, such as insulin and glucagon, did not seem to influence the reduction of hyperglycemia.

 This study was conducted to evaluate the frequency of DNA transfection into human cells following X-ray irradiation. We transfected plasmid DNA (pSV2neo) into human cells, HeLa and PA-1, by either calcium phosphate precipitation or the lipofection method immediately after irradiating the cells with various doses of X-rays. The transfection frequency was evaluated by counting the number of G418-resistant colonies. When circular plasmid DNA was used, irradiation up to a dose of 2 Gy dose-dependently increased the transfection frequency, which reached a maximum of 5 to 10-fold that of the control unirradiated cells. When linear plasmid DNA was used, the transfection frequency was 2 times higher than that of circular DNA. All five of the clones that were randomly chosen expressed the transfected neo gene. In addition, the pSV2neo gene was randomly integrated into the genomic DNA of each clone. These findings indicate that X-ray treatment can facilitate foreign DNA transfer into human cells and that radiation-induced DNA breaks may promote the insertion of foreign DNA into host DNA. The enhancement of DNA transfection with X-rays may be instrumental in practicing gene therapy.

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.