Treatment of chromomycosis with oral high-dose amphotericin B.
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Amphotericin B↗
Biomedical subjects
S Iijima
Publications and source records attributed to S Iijima.
Vitamin D receptor gene polymorphism and bone mineral density in healthy Japanese women.
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Adult↗
Molecular properties, substrate specificity and regulation of DNA-dependent protein kinase from Raji Burkitt's lymphoma cells.
A double-stranded DNA-dependent protein serine/threonine kinase (DNA-PK) was purified from a nuclear extract of Raji Burkitt's lymphoma cells by a three-step column-chromatographic procedure. The main silver-stained band visualized after SDS/PAGE corresponded to an autophosphorylated polypeptide of about 350-kDa that represents the catalytic component. The existence of Ku DNA-binding protein as a regulatory component in the purified enzyme was revealed by Western blot/enzyme immunoassay and direct inhibition test with anti-Ku sera from the autoimmune patients. The DNA-PK catalyzed phosphorylation of synthetic peptides corresponding to Myc and RB proteins in a DNA-dependent manner, indicating that DNA-PK may recognize a second core-sequence motif Pro-Ser/Thr- in addition to the putative consensus sequences of -Ser/Thr-Gln. The level of enzyme activity was significantly higher in DMSO-induced G0/G1-arrested Raji cells as well as in the cells after release from DMSO than in the log-phase cells.
Amino Acid Sequence↗
New pharmacological strategies for cognitive enhancement using a rat model of age-related memory impairment.
We have developed the Stone maze paradigm for use as a rat model of memory impairment observed in normal aging and in Alzheimer's disease. Evidence produced thus far clearly implicates both the cholinergic and glutamatergic systems in acquisition performance in this complex maze task. Although results have been very inconsistent regarding the cognitive enhancing abilities of cholinomimetics for use in Alzheimer's disease, new classes of cholinesterase inhibitors may offer greater therapeutic efficacy. The use of glycine and polyamine agonists appears to be a viable strategy for positive modulation of the NMDA receptor. In addition, an approach that combines stimulation both of cholinergic and glutamatergic systems may have greater potential than agonism of either separately. Manipulation of signal transduction events might also have potential for cognitive enhancement. The influx of Ca2+ through the NMDA receptor stimulates production of NO via the action of NOS. By using NARG to block NOS activity, we have demonstrated in rats that NO production appears to influence learning in the Stone maze. We are currently exploring the age-related changes in NOS activity in specific brain regions of rats to determine if loss in the NO generating system is related to age-related memory impairment observed in the Stone maze. In addition, we are exploring pharmacological strategies for inducing NO production; however, because of the potential neurotoxicity for NO overstimulation, this strategy will present some obstacles. The identification of NO as a simple molecule serving vital physiological functions but representing potential for neurotoxicity presents an important unifying area for neurobiological investigations searching for mechanisms of normal brain aging and of age-related neuropathology, as observed in Alzheimer's disease.
Aging↗
A new runaway type episomal vector for mammalian cells based on a temperature-sensitive simian virus 40 and inducible erythropoietin production.
A runaway vector for mammalian cells was constructed from the simian virus 40 (SV40) genome with a temperature-sensitive mutation of the large T antigen and bacterial neor gene. Replication of this plasmid was repressed above 39 degrees C and vigorous DNA propagation was observed below 33 degrees C in simian CV-1 cells. The human erythropoietin gene was inserted downstream of the SV40 late promoter of the plasmid and the recombinant plasmid was introduced into CV-1 cells. By a temperature shift from 37 to 33 degrees C, the plasmid copy number increased from 5 x 10(2) to 5 x 10(3) copies per cell and the specific production rate of erythropoietin increased more than ten-fold. The bacterial-derived sequences such as the neor gene and vector pUC sequences were prone to delete but the main body of the recombinant plasmid such as SV40 and the erythropoietin-coding sequences were stably maintained at either 33 or 37 degrees C.
Animals↗
Rodent models of memory dysfunction in Alzheimer's disease and normal aging: moving beyond the cholinergic hypothesis.
The Stone maze paradigm has been developed for use as a rat model of memory impairment observed in normal aging and in Alzheimer's disease. Results from several studies have demonstrated the involvement of both cholinergic and glutamatergic systems in acquisition performance in this complex maze task. Although results of clinical studies on the cognitive enhancing abilities of cholinomimetics for treatment of memory impairment in Alzheimer's disease have been inconsistent, new classes of cholinesterase inhibitors offer greater potential for therapeutic efficacy. The physostigimine derivative, phenserine, appears to have marked efficacy for improving learning performance of aged rats or of young rats treated with scopolamine in the Stone maze. Declines in markers of glutamatergic neurotransmission in Alzheimer's disease and in normal aging suggest that pharmacological manipulation of this system might also prove beneficial for cognitive enhancement. Treatment with glycine and/or polyamine agonists is suggested as a strategy for activating the N-methyl-D-aspartate (NMDA) subtype of glutamate receptor. In addition, the use of combined pharmacological activation of cholinergic and glutamatergic systems is suggested. Manipulation of signal transduction events should also be considered as a strategy for cognitive enhancement. The influx of Ca2+ through the channel formed by the NMDA receptor stimulates the production of the oxyradical, nitric oxide (NO*), via the action of nitric oxide synthase (NOS). Compounds that inhibit NOS activity impair acquisition in the Stone maze, suggesting an involvement of NO*. Thus, strategies for inducing NO* production to enhance cognitive performance may be beneficial. Because of the potential neurotoxicity for NO*, this strategy is not straightforward. Although many new directions beyond the cholinergic hypothesis can be suggested, each has its potential benefits which must be weighed against its risks. Nonetheless, an important unifying area for neurobiological research examining mechanisms of normal brain aging and of age-related neuropathology, as observed in Alzheimer's disease, might emerge from the identification of NO* as a simple molecule serving vital physiological functions but representing potential for neurotoxicity.
Aging↗
Elevated chromosome aberration frequency after X-ray exposure of cultured fibroblasts derived from patients with porokeratosis.
Porokeratosis (PK) is a rare genetic skin disorder inherited as an autosomal dominant trait and regarded as a disease predisposing to cancer. To evaluate chromosomal radiosensitivity of PK cells, we examined chromosome aberration frequency after X-irradiation of cultured skin fibroblasts derived from PK patients and controls. Without X-ray exposure, frequencies of chromosome-type aberrations (exchanges or deletions) were not different between the patients and controls. Following X-ray irradiation, frequencies of deletions in the patient group were significantly increased, whereas those of exchanges were not elevated. No differences in chromatid-type aberration frequency were found between the patients and controls with or without exposure to X-ray. The observed radiosensitivity, though not as high as in ataxia telangiectasia (AT) cells, agrees well with the previously reported higher radiosensitivity of PK fibroblasts in survival analysis.
Analysis of Variance↗
Nutrient-induced thermogenesis (NIT) following amino acid infusion.
Nutrient-induced thermogenesis (NIT) induced by parenteral infusion of amino acid (AA) mixtures of different composition and of the same AA mixtures given via different routes (parenteral or intraportal infusion) were investigated in rats using a small animal indirect calorimeter. When 8 different AA solutions of differing composition but with the same total concentration were infused parenterally, both standard NIT (each AA is assumed to generate 3.28 kcal/g) and specific NIT (heat energy of each AA is calculated assuming that it is oxidized to carbon dioxide and water, and metabolised to urea and sulphuric acid) values of the leucine (Leu)-rich and the glycine (Gly)-rich solutions were significantly greater than those of the control solution. Removal of Leu or Gly from the respective AA solutions reversed the increase of both NIT values down to control levels. When the parenteral and portal infusion routes were used in one rat, both NIT values for parenteral infusion of the Leu-rich solution were again significantly greater than those of the control. Likewise, both NIT values for intrportal infusion of the Leu-rich solution were also significantly greater than those of the control. However, no difference in NIT values was found between parenteral and portal infusion of either solution. The result of this study indicated that Leu and Gly may be thermogenic AAs, and the thermogenic effect of Leu is not dependent upon the route of infusion.
Journal Article↗
Evaluation of the thrombolytic effect of tissue-type plasminogen activator with ultrasonic irradiation: in vitro experiment involving assay of the fibrin degradation products from the clot.
Previous studies show that the combined method using thrombolytic drugs and ultrasonic irradiation enhances the effect of thrombolysis in in vitro and in vivo experiments. However, the mechanism of the clot lysis was unclear. In previous studies the evaluation of the clot lysis effect was determined by the rate of decrease in clot weight or the decrease in radioisotope-labeled fibrinogen in the clot. However, it was unclear whether or not the fibrinogen in the clot was degraded. We measured quantitatively the fibrin degradation product, D-dimer (FDP-DD), produced from clots subjected to recombinant tissue-type plasminogen activator (rt-PA, 2.0 micrograms/ml), with and without ultrasonic irradiation. Two levels of continuous ultrasound were used, 300 kHz (100 V, 0.07 W/cm2) and 1 MHz (20 V, 0.4 W/cm2). The quantity of FDP-DD produced was 1.74 times at 300 kHz and 1.80 times at 1 MHz, greater than that measured in rt-PA solution without ultrasonic irradiation; clot lysis was not observed in normal saline with ultrasonic irradiation. Our experiment clarifies that the mechanism of clot lysis when subjected to a combination of drug and ultrasound leads to degradation of fibrin, allowing a quantitative measurement of the enhancement of clot lysis. A high correlation was observed between the FDP-DD produced from the clots and the rate of decrease in clot weight.
Analysis of Variance↗
Development of a thermoelectric cooling apparatus for high-voltage isoelectric focusing on a cellulose acetate membrane.
To develop an isoelectric focusing apparatus using a cellulose acetate membrane (Separax EF), we have designed a thermoelectric cooling isoelectric apparatus. This apparatus has two characteristics. Firstly, the cooling system was switched to a thermoelectric cooling system from an ice-cooling system. Secondly, the chamber lid of the electrophoretic apparatus was also devised so that samples could be applied without opening the chamber lid. With this apparatus we could perform the isoelectric focusing without worrying about room temperature and humidity in the laboratory. Applying 2000 V for an extra 5 min with our module cooling system, we achieved a much higher degree of resolution with three sheets of cellulose acetate membrane (Separax EF) overlaid for simultaneous electrophoresis. Thus, three types of information could be obtained from only one electrophoretic procedure.
Biocompatible Materials↗
[Study on the association between risk factors for osteoporosis and bone mineral density].
The relationship between bone mineral density (BMD) and risk factors for osteoporosis was investigated on 146 females (23-68 years old) in Enzan city, Yamanashi prefecture. BMD was measured by DEXA (Dual Energy X-ray Absorptiometry) method. Information regarding menstruation, health status, life-style, quantities of nutrient intake and frequencies of food intake were obtained by questionnaire. Fitness level was determined by exercise test and daily energy expenditure by calorie-counter. The results were as follows; 1) No relationship between physical activities or daily energy consumption and BMD was found. 2) No relationship between calcium intake or vitamin D intake and BMD was found, but a significant and positive correlation coefficient between intake frequency of milk products and BMD was observed. 3) Significant influences of age, BMI, presence of lumbago and food frequency score on BMD were indicated by multiple regression analysis. Food frequency score were defined as values weighted in each of three food category, obtained by factor analysis. 4) Significant influences of height decrement rate per year and number of years postmenopausal on BMD reduction rate per year were indicated by multiple regression analysis. These results suggest that these significant factors are useful in indications of risk for osteoporosis.
Adult↗
[Involvement of cytokines on muscle protein breakdown under stress: establishment of a new experimental system using C2C12 myotubes: preliminary report].
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Animals↗
Role of intracellular proteases in differentiation of L6 myoblast cells.
The activities of cathepsins B, L, and H increased by 3.0, 2.3, and 1.9 fold in differentiation of L6 myoblasts. Separating from the intrinsic inhibitor, both mu- and m-calpain activities also increased by 4.9 and 1.7 fold during differentiation. Calpeptin (10(-5)M), cell penetrating calpain inhibitor, blocked fusion of myoblast by 55% and increment of creatine phosphokinase (CPK) activity by 20.3% with inhibiting the activities of cathepsin L and mu-calpain. E-64(10(-4)M) did not block fusion of myoblasts but increment of CPK activity by 25.0% with inhibiting the activities of cathepsin L and B. Calpain may be involved in the process of myoblast fusion.
Animals↗
[Analysis of factors for the development of pulmonary tuberculosis in persons with collagenosis].
Recently, the annual reduction in the incidence rate of tuberculosis has slowed in Japan. One reason for this trend is believed to be due to an increase in the number of immunocompromised hosts. In our department, we discovered 10 cases of pulmonary tuberculosis among 962 cases of collagenosis, and have analyzed the factors for the development of pulmonary tuberculosis in these patients. A total of 29 patients wer involved in the study: 22 with systemic lupus erythematosus (SLE) (active disease, 10; inactive disease, 12) and having no pulmonary complications, and seven with pulmonary tuberculosis and no concomitant diseases. Our findings were as follows: 1. Seven of the 10 patients with pulmonary tuberculosis also suffered from SLE. 2. Nine of the 10 patients had been treated with a corticosteroid or immunosuppressant. 3. Serum CH50 and erythrocyte sedimentation rate (ESR) were valuable indicators for diagnosing pulmonary tuberculosis in the patients with SLE. 4. Patients with SLE and pulmonary tuberculosis tended to show lymphopenia in peripheral blood, which was further enhanced by prolonged use of steroids. 5. Miliary tuberculosis rather than pulmonary tuberculosis tended to develop in elderly patients, and required longer hospitalization.
Adrenal Cortex Hormones↗
Expression of active human DNA ligase I in Escherichia coli cells that harbor a full-length DNA ligase I cDNA construct.
A recombinant plasmid for expression of full-length human DNA ligase I (phLig-I) was constructed in a plasmid/phage chimeric vector, pTD-T7N, which was derived from pUC118 by oligonucleotide-directed mutagenesis. The insert contained a 2757-base pair coding sequence for a whole human DNA ligase I and an extra ACC codon adjacent to the ATG initiation codon. This ACC codon was required for achieving high levels of expression of full-length DNA ligase I in Escherichia coli strain BL21. The recombinant plasmid, which was designed to exploit the T7 late promoter and the ATG initiation codon for beta-galactosidase was transfected into E. coli BL21 cells that express T7 RNA polymerase. The recombinant clone produced relatively high levels of DNA ligase I with a molecular mass of 130 kDa, as estimated by SDS-polyacrylamide gel electrophoresis. The DNA ligase was purified to near-homogeneity by the two-step column chromatographic procedure from BLphLig-I cells that had been induced with isopropyl beta-D-thiogalactoside. The specific activity, chromatographic behavior, kinetic properties, molecular mass, and antigenicity of the recombinant human DNA ligase I were indistinguishable from those of purified mammalian DNA ligase I. Metabolically labeling experiments with 32P(i) indicate that the recombinant DNA ligase I was present as an enzyme-AMP reaction intermediate, but not as a phosphoprotein, in the E. coli cells.
Amino Acid Sequence↗
Electron-energy-loss spectroscopy of carbon nanometer-size tubes.
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Journal Article↗
Immunologic significance of increased soluble CD8/CD4 molecules in patients with active systemic lupus erythematosus.
This study attempted to estimate soluble CD4(sCD4)/CD8(sCD8) molecules in active systemic lupus erythematosus (SLE) patients. Measurements were made by solid-phase enzyme-linked immunosorbent assay. sCD8 or sCD4 molecules were significantly increased in the patients as compared to control subjects. sCD8 correlated with the erythrocyte sedimentation rate. sCD4 correlated with the anti DNA antibody titer, the IgG concentration, and negatively with the complement titer. An association of these molecules with immunologic abnormalities and disease activity exists in SLE patients.
CD4 Antigens↗
Effect of indomethacin on postoperative protein metabolism after gastrectomy under total parenteral nutrition.
A randomized trial was undertaken to evaluate the effects of postoperative indomethacin (IDM) administration on protein metabolism in 20 patients who underwent an uncomplicated distal gastrectomy and were placed on post-operative total parenteral nutrition (TPN). Ten patients (the IDM group) received 50 mg of IDM every 8 h after operation up to postoperative day (POD) 4 while the other ten patients (the control group) received neither IDM nor any other non-steroidal anti-inflammatory drug, postoperatively. Though the requirement for postoperative plasma transfusion was significantly greater in the IDM group, the albumin level on POD 1 was significantly lower in this group than in the control group. The postoperative changes of C-reactive protein, retinol binding protein, and pre-albumin between the two groups showed no difference. Moreover, the urinary 3-methylhistidine excretion, N-balance, and plasma aminogram on POD 4 also demonstrated no difference. We thus concluded that post-operative IDM administration after elective surgery has no additional anti-catabolic effect on the presence of TPN.
Blood Proteins↗