Effects of the Addition of Beta2-agonist Tulobuterol Patches to Inhaled Corticosteroid in Patients with Asthma

ABSTRACTBackgroundWhether the additive effects of the tulobuterol patch (TP), the world’s first transdermal beta2-agonist preparation, are useful in asthma patients receiving inhaled corticosteroid (ICS) is unclear. To examine the add-on effects of TP on bronchial hyperresponsiveness and reduction of the percentage of sputum eosinophils, and to compare add-on effects of TP, slow-release theophylline (SRT), and a leukotriene receptor antagonist (LTRA) in patients with asthma receiving ICS.MethodsStudy 1: We randomly allocated 24 patients with asthma receiving ICS alone in equal numbers to either control treatment (ICS alone at conventional doses) or TP treatment (ICS at conventional doses plus TP at 2 mg/day). Following a 2-week observation period, patients received the allocated drug regimens for 4 weeks. Methacholine challenge test and measurement of percentage of eosinophils in hypertonic saline- induced sputum were performed before and after the treatment period. Study 2: We compared add-on effects of TP, SRT, and LTRA in 65 patients with asthma receiving ICS alone, using spirometry and peak expiratory flow (PEF). Participants in these studies had experienced decrease in morning PEF to <80% of the predicted value at least twice a week.ResultsStudy 1: In the TP group, improvement of bronchial hyperresponsiveness and decrease in percentage of sputum eosinophils both indicated a statistically significant difference (p < 0.01, and p < 0.05, respectively). These findings were not observed in the control group. Study 2: forced expiratory volume in 1 second (FEV1) and PEF markedly increased after treatment with TP compared with treatment with SRT or LTRA.ConclusionsThese findings suggest that TP can be used as a long-term add-on controller for patients with asthma receiving ICS

Function of Monocytes in the Retired Workers of the Okunojima Poison Gas Factory

Monocyte function of poison gas workers was determined and the following results were obtained. 1) No difference in cytostatic activity could be observed between poison gas workers and their controls. 2) Phagocytic activity of poison gas workers was slightly depressed when compared with that of the controls, but the difference was not significant. By duration of work, it was observed that the group with duration of work exceeding five years had a significantly lower value when compared with the group with duration of work being less than two years. 3) Chemotactic activity of poison gas workers tended to be depressed when compared with that of the controls. The activity tended to be more depressed the longer the duration of work. 4) A significant positive correlation was observed between cytostatic activity and phagocytic activity

A Novel Method for Screening Monoclonal Antibodies Reacting with Antigenic Determinants on Soluble Antigens; A Reversed Indirect-Enzyme Linked Immunosorbent Assay(RI-ELISA)

A novel screening method was established to select new monoclonal antibodies which react with unknown antigenic determinants on molecules bearing antigen determinants reactive with established monoclonal antibodies. This new method is a sandwich assay termed "reversed indirect-enzyme linked immunosorbent assay" (RI-ELISA). Goat antimouse immunoglobulin antibodies are used as the primary immobilized antibody in this assay. They allow the non-purified monoclonal antibodies contained in hybridoma culture supernatants to bind to the microtest plate for enzyme immunoassay (EIA plate) much more efficiently than in the usual sandwich assay where the non-purified monoclonal antibodies are adsorbed directly to the polystyrene surface. The antigen solution is then reacted with the monoclonal antibodies and thereafter enzyme labeled monoclonal antibody with known specificity is added. Therefore, if the hybridoma culture supernatant contains monoclonal antibodies which were bound to the EIA plate and react with antigenic determinants on the soluble molecules which have antigen determinants recognized by the enzyme labeled antibody, the enzyme labeled antibodies will bind to induce an enzymatic reaction. The most important technical consideration in the RI-ELISA is the inhibition of direct binding of the enzyme labeled monoclonal antibodies to free sites remaining in the immobilized goat anti-mouse immunoglobulins antibodies. This problem could be effectively overcome by using normal mouse serum as blocking substance. These studies indicate that the RI-ELISA may be a useful screening method for selecting new monoclonal antibodies which react with unknown antigenic determinants on soluble molecules

Study of Genetic Effects of Sulphur Mustard Gas on Former Workers of Ohkunojima Poison Gas Factory and Their Offspring

General health examination and one-dimensional electrophoretic examination to detect mutations at the protein level were conducted in order to elucidate potential genetic effects of sulphur mustard gas on children of the former workers of Ohkunojima Poison Gas Factory. In the general examination, no disease which was definitely considered to be caused by genetic effects was observed, and no examination values obtained for the children proved to be significantly abnormal compared with those for their parents. Blood samples from 456 children were electrophoretically examined for 30 protein systems. A total of 36 protein variants were detected in 10 protein systems, and the frequency of variants was 2.63 per 1,000 tests. Family study was completed for 32 of these variants, all of which were confirmed to be genetic variants. In 29,868 locus tests, mutation occurred in germ cells of parents could not be detected. Among 36 variants, two PGM2 variants and one GPI variant were detected for the first time in this study

Serum and Urinary Type IV Collagen Concentrations in the Assessment of Diabetic Microangiopathy

We investigated the role of measurement of serum and urinary type IV collagen (IV-C) levels in monitoring diabetic microangiopathy. Furthermore, we compared these levels in diabetic nephropathy and non-diabetic renal disease (NDRD). A one-step sandwich enzyme immunoassay was used to measure IV-C levels in 82 diabetic patients, 33 NDRD patients and 20 healthy non-diabetic control subjects. The diabetic patients were classified into four groups according to urinary albumin I creatinine index CACI) (mg/g) and serum creatinine (s-Cr) (mg/dl) : normoalbuminuria (ACI300, s-Crl.99 mg/dl). Serum and urinary IV-C levels were significantly elevated even in diabetic patients without clinical evidence of microangiopathy compared with control subjects (p<0.05 and p<0.01, respectively). Both levels were significantly higher in normoalbuminuric patients than in the control subjects, and in patients with microalbuminuria, albuminuria or renal insufficiency than in normoalbuminuric patients, with significant differences between these groups (serum and urinary IV-C, both p<0.0001 by ANOVA). Urinary IV-C and albumin levels were significantly correlated, even in normo- and microalbuminuric patients (r = 0.55, p<0.0001). Serum IV-C in normoalbuminuric patients rose significantly as the degree of retinopathy progressed from background to proliferative stages (p<0.05). Neither serum nor urinary IV-C levels were influenced by glycemic control. Albuminuric diabetic patients (with and without renal insufficiency) had significantly higher levels of serum IV-C compared with those in proteinuric NDRD patients (p<0.005), though there was no significant difference in the urinary IV-C level. However, the urinary IV-CI albumin ratio was significantly higher in albuminuric diabetic patients than in proteinuric NDRD patients, even after adjusting for s-Cr and creatinine clearance (p<0.0001). In conclusion, we suggest that measured serum and urinary IV-C concentrations may serve as new markers for monitoring the development and progression of diabetic microangiopathy, particularly nephropathy. Furthermore, the measurement of serum IV-C concentrations and urinary IV-CI albumin ratios in diabetic patients may allow diabetic nephropathy and non-diabetic renal disease to be differentiated.A part of this work was presented at the 36th annual meeting of the Japan Diabetes Society (Sendai, Japan, May 13-15, 1993), and another part has been accepted for presentation at the 15th International Diabetes Federation Congress (Kobe, Japan, November 6-11, 1994)

Production of Interleukin 2 in Human Peripheral Blood Lymphocytes: Optimal Condition for its Culture

For the assay of the production of Interleukin 2 (IL-2) in human peripheral blood lymphocytes (human PBL), a study was made on the optimal condition for its culture. 1) The optimal condition for the production of IL-2 was considered to be incubation time of 24 hr, number of PBL of 1 x 106 cells/ml, and phytohemagglutinin-M (PHA-M) concentration of 1 — 5% 2) By one way mixed lymphocyte reaction (one way MLR), IL-2 activity similar in level to that by PHA-M stimulation could also be obtained. It was maximal at the 4th day of culture. 3) By PHA-P (0.06%) stimulation, IL-2 could also be produced similar to that by PHAM stimulation but it decreased in the order of Con A, PWM and PPD and hardly any production of IL-2 could be observed by LPS

Interleukin-2 Production and Lymphocyte Proliferation in Aged and Young Humans

A study was made on the effect of aging on the production of Interleukin-2 (IL-2) and lymphocyte proliferation of human peripheral blood lymphocytes. 1) In comparison with young individuals, IL-2 production tended to decrease in aged individuals, while lymphocyte proliferation showed a significant decrease. 2) A significant correlation was observed between IL-2 production and lymphocyte proliferation in both the aged and young human populations. 3) IL-2 production showed a negative correlation with Leu-2a positive rate and a positive correlation with Leu-3a/Leu-2a ratio in aged individuals

Detection of Lymphocyte Subsets by Monoclonal Antibodies in Aged and Young Humans

With the use of Leu-series monoclonal antibodies, peripheral blood lymphocyte subsets in aged and young humans were determined. 1) In comparison with young individuals, Leu-1+ cells and Leu-2a+ cells were decreased, whereas Leu-7+ cells and Leu-3a/Leu-2a were increased in aged individuals. 2) No sex difference could be observed in lymphocyte subsets. 3) PHA response of lymphocytes showed a negative correlation with Leu-2a+ cells and a positive correlation with Leu-3a/Leu-2a in aged individuals

Immune Functions of Former Poison Gas Workers I. Mitogenic response of lymphocytes and serum factors

The relation of depressed immune function to carcinogenesis has been estimated in the living body. The authors have measured the immunological parameters in former poison gas workers, a group having a high risk of carcinogenesis, for comparison with age matched normal controls and the following results were obtained. 1) With regard to serum factors, no significant difference could be demonstrated between normal controls and poison gas workers in such immunoglobulins as IgG, IgA, and IgM, in acute phase reactants such as α1-AT, α1-AG, α2-HS and C3 and in such tumor markers as CEA, ferritin, and β2-microglobulin. Furthermore, no difference could be observed in the positive rate of immune complex and in complement activity. 2) No difference could be observed between the two groups with regard to tuberculin skin reaction and number of lymphocytes, but the longer the duration of work at the poison gas factory, the more significant was the increase in those who showed negative tuberculin skin reaction. 3) In comparison with normal controls, mitogenic response to PHA showed a significant decrease in poison gas workers, but no significant difference could be seen in mitogenic response to Con A and PPD and in mixed lymphocyte reaction. 4) No significant difference could be demonstrated between the two groups in the inhibitory effects of serum on mitogenic response to PHA and Con A and on mixed lymphocyte reaction

Comparison of Various Serum Protein Values in the Japanese and the Japanese-Americans Resident in the United States

Measurements were made of various types of proteins, that is α1-antitrypsin, α1-acid glycoprotein, α2-HS glycoprotein, haptoglobin, α2-macroglobulin, transferrin, C3, IgG, IgA and lgM, in the serum of the Japanese-Americans living in Hawaii and the Japanese-Americans living in Los Angeles who are assumed to be genetically almost identical to the Japanese in Hiroshima Prefecture but are known to have a higher intake of animal fats but a lower intake of complex carbohydrates. These were compared with those of the Japanese in Hiroshima Prefecture. α2-macroglobulin values in serum of the male Japanese-Americans living in Hawaii of ages 30-39 years, 40-49 years, and 50-59 years were significantly lower than those of the residents in Hiroshima Prefecture, but no significant difference in these values could be observed between the Japanese-Americans living in Los Angeles and the Japanese in Hiroshima Prefecture. No significant difference could be observed in the values of other serum proteins in all age groups. These findings indicate that the difference in intake volume of animal fats and complex carbohydrates did not affect these serum protein values