Electron density measurement of cesium seeded negative ion source by surface wave probe.

Electron density measurements of a large-scaled negative ion source were carried out with a surface wave probe. By comparison of the electron densities determined with the surface wave probe and a Langmuir probe, it was confirmed that the surface wave probe is highly available for diagnostic of the electron density in H(-) ion sources. In addition, it was found that the ratio of the electron density to the H(-) ion density dramatically decreases with increase of a bias voltage and the H(-) ions become dominant negative particles at the bias voltage of more than 6 V.

Spatial distribution of the charged particles and potentials during beam extraction in a negative-ion source.

We report on the characteristics of the electronegative plasma in a large-scale hydrogen negative ion (H(-)) source. The measurement has been made with a time-resolved Langmuir probe installed in the beam extraction region. The H(-) density is monitored with a cavity ring-down system to identify the electrons in the negative charges. The electron-saturation current decreases rapidly after starting to seed Cs, and ion-ion plasma is observed in the extraction region. The H(-) density steps down during the beam extraction and the electron density jumps up correspondingly. The time integral of the decreasing H(-) charge density agrees well with the electron charge collected with the probe. The agreement of the charges is interpreted to indicate that the H(-) density decreasing at the beam extraction is compensated by the electrons diffusing from the driver region. In the plasmas with very low electron density, the pre-sheath of the extraction field penetrates deeply inside the plasmas. That is because the shielding length in those plasmas is longer than that in the usual electron-ion plasmas, and furthermore the electrons are suppressed to diffuse to the extraction region due to the strong magnetic field.

Characteristics of 80 keV positive ion source for Large Helical Device.

An additional beamline, BL5, equipped with four positive ion sources will be installed on Large Helical Device (LHD) in 2010. The performance of an ion source which generates 80 keV deuterium and 60 keV hydrogen beams was investigated. The structure of the ion source is based on that of a BL4 ion source on LHD. The main differences between the ion sources for the BL4 and BL5 are the acceleration voltages and the materials of plasma electrodes: copper and molybdenum, respectively. The molybdenum plasma electrode for BL5 has better performance than the copper plasma electrode of BL4. The integrated performance of the ion source for BL5 reached a value equivalent to approximately 58 A in the beam current of hydrogen positive ion at 60 keV in the beam energy.

Beamlet characteristics in the accelerator with multislot grounded grid.

Characteristics of multibeamlets are investigated by means of beamlet monitoring technique. The beamlets are extracted from an accelerator with multislot grounded grid and the profiles are observed as infrared images of temperature distributions on a cold isostatic pressed graphite plate exposed by H-beamlets. The optimal horizontal and vertical divergence angles of single beamlet are estimated at 4.1 and 6.1 mrad, respectively.

Spectroscopic observations of beam and source plasma light and testing Cs-deposition monitor in the large area negative ion source for LHD-NBI.

In the large area negative ion source for the LHD negative-ion-(H(-))-based neutral beam system, (I) we used the spectrometer to measure caesium lines in the source plasma during beam shots. (II) With Doppler-shifted measurements, the H(alpha) line at three different locations along the beam as well as the spectrum profile for cases of different plasma grid areas. (III) Caesium deposition monitor with a high speed shutter was tested to measure the weight of the deposited Cs layer. In the observation, cleaner spectra of Doppler-shifted H(alpha) line with only a small level of background light were obtained at a new observation port which viewed the blueshifted light in the drift region after the accelerator of a LHD ion source. Both the amounts of Cs I (852 nm, neutral Cs(0)) and Cs II (522 nm, Cs(+)) in the source plasma light rose sharply when beam acceleration began, and continued rising during a 10 s pulse. It was thought that this was because the cesium was evaporated/sputtered from the source back plate by the back-streaming positive ions. Cs deposition rate to the crystal sensor measured by adjusting the shutter open time was evaluated to be 2.9 nanograms/s cm(2) for preliminary testing. More neutral Cs tended to be evolved in the source after arc discharge. Much Cs could be consumed in a high rate-pulsed operation (such as LHD source).

Neutral beam injection with an improved accelerator for LHD.

The beam profiles, port-through, rates and injection powers obtained with an improved accelerator with the multislot grounded grid are described. The accelerator has a combination of a steering grid with racetrack shaped aperture and multislot grounded grid to improve the beam optics. The optimal beam optics is obtained at the voltage ratio of 16.5-16.8, and the profiles are well fit by superposing multibeamlets with the divergent angles of 5.0 and 7.2 mrad along the direction parallel to the long and short axes of the slots of grounded grid. By adopting the racetrack shaped steering grid, the port-through rate increases from 34% to 38%, and the maximum injection power reaches 6 MW/187 keV.

Vegetative pyoderma gangrenosum in chronic renal failure.

We report the case of a 58-year-old woman with painful recurrent erythematous lesions on both legs of 6 months' duration. The patient had been treated with haemodialysis for chronic renal failure for the previous 10 years. Physical examination revealed pea-sized erythematous nodules with sinus formation and discharge of bloody pus, which yielded neither bacterial nor fungal cells in culture. These lesions healed leaving cribriform scars, after bed rest, topical disinfection and systemic and in some areas intralesional corticosteroids. Cases of pyoderma gangrenosum are usually classified as one of four clinical variants: ulcerative, pustular, bullous and vegetative. Our case might correspond to a vegetative form in view of the development of sinuses, neutrophilic abscesses and palisading granuloma, even though no ulceration was observed.

A case of pencil core granuloma with an unusual temporal profile.

The patient is a 47-year-old female with a bluish tumor resembling malignant melanoma at the macroscopic level on the medial aspect of her left big toe. The patient had crushed a pencil with this toe about 30 years previously and, since then, a bluish lesion had been present. About 15 years ago, the lesion had suddenly increased in size over the course of a few months. Subsequently, the size of the lesion had not changed noticeably. Ultrasonography, but not magnetic resonance imaging was a helpful preoperative examination to distinguish the lesion from malignant melanoma. The excised lesion contained a piece of material that resembled pencil lead and bluish mud. X-ray microanalysis of the lead-like material revealed that its composition was similar to that of pencil lead. Histologic examination showed features of foreign-body reaction, except for necrotic change and few histiocytes.

The complete amino acid substitutions at position 131 that are positively involved in cold adaptation of subtilisin BPN'.

To ascertain whether position 131 of a mesophilic protease, subtilisin BPN', is a potential critical site for cold adaptation as screened by evolutionary engineering (S. Taguchi, A. Ozaki, and H. Momose, Appl. Environ. Microbiol. 64:492-495, 1998), a full set of subtilisin BPN' mutants with mutations at position 131 was constructed by site-saturation mutagenesis. All mutated enzymes were measured for specific activity at 10 degrees C by the quantitative titer microplate assay system using polyclonal antibody against subtilisin BPN' and a synthetic chromogenic substrate. All the mutants exhibited proteolytic activities almost the same as or higher than that of the wild-type enzyme, suggesting that position 131 may be important for cold adaptation. In comparison with the wild type, purified mutants G131F, G131R, G131M, and G131W were found to acquire proteolytic activities (k(cat)/K(m)) at 10 degrees C that were 150, 94, 84, and 50% higher, respectively. In particular, for the G131F mutant, temperature dependency in enzyme activity was shown by an increase in k(cat) and a decrease in K(m). All of these amino acid substitution mutants, G131F, G131R, G131M, and G131W, acquired increased proteolytic activities at 10 degrees C for three different synthetic peptide substrates but no increase in caseinolytic activity. Furthermore, they all conferred thermolability on the enzyme to differing extents in terms of the half-life of enzyme inactivation at 60 degrees C. No significant correlation was found between the amino acids preferred for cold adaptation surveyed here and those present at position 131 of subtilisin of psychrophilic cells naturally occurring in cold environments. Based on these findings, position 131 is a contributor in artificial evolution for acquiring a cold-active character and may not be related to physiological requirements for subtilisin-producing cells living in cold environments. Therefore, saturation mutagenesis would be effective in achieving rapid improvement in protein properties via evolutionary engineering.

Quantitative assay system for specific enzyme activity using antibody: the case of protease, subtilisin BPN'.

Assay on a high-quality microtiter plate was found to allow for quantitative analysis of bacterial serine protease, subtilisin BPN', and its mutant enzymes which had been genetically engineered to be adapted to low-temperatures (Taguchi et al., 1998. Appl. Environ. Microbiol. 64, 492-495), by using polyclonal antibody against subtilisin BPN'. The use of polyclonal antibody was crucial in normalizing the number of various different enzyme molecules in culture supernatant samples of recombinant strains of Bacillus subtilis, giving rise to the performance of specific activity assay of the enzymes. Relative activity of each mutant subtilisin BPN' to wild-type enzyme was estimated by monitoring the increased value of absorbance caused by enzymatic hydrolysis of a chromogenic substrate. The relative activity in each enzyme estimated by this method showed good coincidence with that estimated by kinetic parameters, kcat/K(m) of the purified enzymes. We termed the system as 'ABEA' (antibody-bound enzyme assay). The efficient ABEA system developed here would be useful for the determination of specific activity of other enzymes of interest and provide us versatile applications in the field of evolutionary engineering.

[A case report of mild from of phenylketonuria].

We report a 19-year-old man with mild form of phenylketonuria. The diagnosis was first made when he was examined for the tremor at 19 years of age. He had not received the Guthrie's screening test for phenylketonuria in infancy. His development of speech and walking was almost normal. Action and positional tremor developed at two years of age, and psychomotor deterioration at five years. His intelligence was of borderline, and he entered the special class for the mentally retarded at elementary school and junior high school. His skin and iris were less pigmented than those of Japanese young adults, and his hair was rather reddish. He had mild action tremor. He showed mild mental retardation, and the WAIS was 46 in PIQ, 70 in VIQ and 53 in total IQ. T2-weighted MRI of the brain showed high signal of the deep white matter around the posterior conus of the lateral ventricles. EEG showed paroxysmal abnormalities. Serum aminogram disclosed a marked elevation of phenylalanine. Analyses of pteridine in the serum and urine disclosed a low ratio of neopterine/biopterine. An assay of the dehydropteridine reductase in erythrocytes was normal. These laboratory data indicated that his condition was caused by a deficiency of phenylalanine hydroxylase deficiency.

Tenascin expression and postnatal development of the human prostate.

We investigated the distribution of tenascin during postnatal development of the human prostate. Monoclonal antibody specific to human tenascin was applied to paraffin sections by the avidin-biotin-complex method to examine its localization. Infantile prostates showed two distinct zones. The inner zone had sparse acini with fibromuscular bundles. The peripheral zone had similar acini and lighter stroma as compared with inner zone. Tenascin was found diffusely but weakly. The periglandular area occasionally showed immunoreactivity. The prostates from 9- to 13-year-old subjects had a morphology similar to that of the infantile gland. The difference was increased density of the acini. Immunoreactivity was low. In the prostates from 14- to 21-year-old subjects, the acini distribution was more crowded, and the epithelial lining had become taller in both zones. Tenascin distributed preferentially in the peripheral zone during this period. Simultaneously, the percent of glandular area in the peripheral zone rose abruptly. The dynamics of tenascin expression are closely associated with the development and maturation of the gland. The distribution of tenascin during the post-puberal period may suggest its participation in the preferential of prostatic carcinoma in the peripheral zone.

Intranuclear androgen concentrations in facial skin.

The concentrations of 5 alpha-dihydrotestosterone (DHT) and testosterone were measured by radioimmunoassay (RIA) in the crude nuclear and cytoplasmic fractions of facial skin containing large sebaceous glands. The data obtained were compared with those obtained with specimens from non-target areas. The intranuclear levels of DHT and testosterone in facial skin were 0.03 +/- 0.3 pg/micrograms DNA and 0.35 +/- 0.03 pg/micrograms DNA, respectively levels in genital skin. By contrast, both androgens were below detection by RIA in non-target skin. There was no significant difference between men and women with respect to the intranuclear androgen levels in facial skin. These findings support the view that the sebaceous gland is a typical androgen target organ irrespective of sex.

Intranuclear androgen and cytosolic receptor concentrations in the axillary skin of osmidrosis.

5 alpha-Dihydrotestosterone (DHT) and testosterone were measured by radioimmunoassay in the crude nuclear and cytoplasmic fractions of the axillary skin of both male and female patients with osmidrosis and the levels compared with those of nongenital skin. The intranuclear levels of DHT were 1.44 +/- 0.22 and 1.77 +/- 0.38 pg/micrograms DNA in men and women, respectively. Those of testosterone were about 10% of DHT levels. In the skin of nontarget regions nuclear DHT was much scarcer or undetectable. Cytosolic androgen receptors in isolated apocrine glands were also measured using 3H-R1881 as a ligand. Typical androgen receptors were present in all of eight patients (KD = 1.32 +/- 0.24 X 10(-9)M, Bmax = 10.3 +/- 0.51 fmol/mg protein). Neither the intranuclear androgen concentrations nor the cytosolic androgen receptor levels were significantly different between the two sexes. These data indicate clearly that the apocrine gland of patients with osmidrosis is a typical androgen target organ, irrespective of sex, and suggest that nuclear DHT in the axillary skin of women is derived from not only testosterone but also other precursors, especially in consideration of the very low serum concentrations of testosterone in females.

Side gland of Suncus murinus as a new model of sebaceous gland: 5 alpha-reductase, androgen receptor, and nuclear androgen content in male and female animals.

The side gland of Suncus murinus is composed of well-developed sebaceous glands in both males and females. We measured the 5 alpha-reductase activity, androgen receptor content, and the intranuclear concentrations of testosterone and dihydrotestosterone in this side gland taking it as a new experimental model of human sebaceous glands. Lineweaver-Burk plot analysis suggested the presence of two classes of 5 alpha-reductase with different Km for testosterone in the homogenate. The enzyme activity was slightly higher in males than in females in the presence of a high concentration of testosterone. The levels of androgen receptors were approximately 40 and 30 fmol/mg protein in the cytosol and nuclei, respectively. The values did not differ significantly between males and females. The intranuclear concentration of dihydrotestosterone in the side gland was higher than that of testosterone in each sex. The intranuclear level of each of these two androgens in the female side gland was comparable to that in the male side gland despite the fact that the serum level of testosterone was much lower in the female. These data clearly indicate that the side gland is a typical target tissue for androgens in the female as well as in the male. Androgens other than testosterone may serve as precursors of dihydrotestosterone in the female side gland.

Estrogen binding component in the pilosebaceous tumor developed in Suncus murinus.

We transplanted a pilosebaceous tumor developed on the sidegland of Suncus murinus to male nude athymic (BALB/c-nu/nu) mice. This tumor can be transplanted to female hosts as well, with a lower rate of graft-taking and slower growth rate. In this study we demonstrated the presence of macromolecules which specifically bind to estrogen. Measurement of 17 beta-estradiol (E2) binding by a dextran-coated charcoal assay revealed that the number of binding sites and the dissociation constant were 22.3 +/- 4.6 fmol/mg protein and 1.4 +/- 0.24 X 10(-9) M, respectively. This binding was specific for E2 and diethylstilbestrol (DES). Sucrose gradient centrifugation of the [3H]E2-labeled cytosol yielded a sharp peak of radioactivity at 3.5S-4S under high salt conditions and a 9S peak with a shoulder at 3.5S under low salt conditions. This 3.5S shoulder was due to dissociation of [3H]E2 from the 9S peak during the centrifugation, since only the 9S peak was obtained by postlabeled density gradient analysis. An assay of the in vivo binding of [3H]E2 showed significant radioactivity in the nuclear extract from the tumor. This nuclear uptake was markedly decreased by simultaneous administration of 100-fold excess of E2. In tumor-bearing castrated nude mice, 1-100 micrograms/day of E2 did not affect tumor growth, whereas it counteracted the stimulative effect of testosterone propionate.

Transcatheter arterial embolization of vesical artery in the treatment of invasive bladder cancer.

Transcatheter arterial embolization was performed in a total of 70 patients with invasive bladder cancer from October 1980 to December 1987. It was performed as a part of multidisciplinary treatment in patients with invasive bladder cancer. Microencapsulated mitomycin C, gelatin sponge and lipiodol (iodized oil) were used as the embolic material. As a result, reduction of tumor size was noted in 56.7%, but there was no embolic materials-related significant difference. Stage reduction was noted in 72.7% of patients where pathological stages were confirmed by operation. Hemostatic effects were noted in 76.5%. The symptoms of side effects included fever, leukocytosis, urinary frequency and pain, but none of them was severe. The results of the present study indicate that embolization of the vesical artery is useful in the treatment of bladder cancer.

[Usefulness of computed tomography and ultrasonography for the early detection of renal cell carcinoma].

Eighty-three cases of renal cell carcinomas admitted to Nara Medical University and its related hospitals from August, 1962 through July, 1984 were reviewed. We have been using computed tomography (CT) and ultrasonography (US) for early detection of renal cell carcinomas since 1980. Thereafter the number of patients with low stage renal cell carcinoma was significantly increased. Furthermore 6 carcinomas were incidentally detected by CT and/or US examination for checkup of other diseases. We believe that CT and US may be valuable as a screening modality for early detection of renal cell carcinoma.

[Clinical analysis of primary vesicoureteral reflux in children].

The retrospective analysis of 64 patients between 1 month and 14 years old with 87 primary vesicoureteral units with reflux was reviewed. They were followed-up for a period of 3 to 60 months. Thirty one patients were managed conservatively by chemotherapy (group A) and 28 patients were treated surgically. As the anti-reflux operation the Politano-Leadbetter procedure was used in 27 patients and the modified Politano-Leadbetter procedure in 1 patient (group B). Five patients were not treated (group C). In 13 of the 30 renal units of group A, the vesico-ureteral reflux disappeared spontaneously and in group B the post-operative cystourethrography revealed cure of reflux in all the renal units. In 2 of the 21 renal units of group A excretory urography (IVP) revealed improvement and in 3 deterioration. Post-operative IVP revealed improvement in 13 of 39 renal units of group B and in 2 deterioration. In 3 of the 22 renal units of group A the renogram revealed improvement and in 3 deterioration. The post-operative renogram revealed improvement in 8 of the 28 renal units of group B and deterioration in renal units. Analysis was also made according to the vesicoureteral reflux grade.