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Biomedical subjects

K Tokita

Publications and source records attributed to K Tokita.

At least 19 recordsLinked to original sources

Accumulation of cyclic ADP-ribose measured by a specific radioimmunoassay in differentiated human leukemic HL-60 cells with all-trans-retinoic acid.

Cyclic adenosine diphosphoribose (cADPR) is a novel candidate for the mediator of Ca2+ release from intracellular Ca2+ stores. The formation of this cyclic nucleotide is catalyzed by not only Aplysia ADP-ribosyl cyclase but also an ecto-form enzyme of NAD+ glycohydrolase (NADase), which was previously identified as all-trans-retinoic acid (RA)-inducible CD38 in human leukemic HL-60 cells. In the present study, we developed a radioimmunoassay specific for cADPR, by which more than 100 fmol of cADPR could be detected without any interference by other nucleotides. The possible involvement of CD38 in the formation of cellular cADPR was investigated with the radioimmunoassay method. A marked increase in cellular cADPR was accompanied by all-trans-RA-induced differentiation of HL-60 cells. Moreover, a high level of cellular cADPR was observed in other leukemic cell lines, in which CD38 mRNA was expressed. Thus, CD38, which was initially identified as an NADase, appeared to be responsible for the formation of cellular cADPR.

ADP-ribosyl Cyclase

Enzyme properties of Aplysia ADP-ribosyl cyclase: comparison with NAD glycohydrolase of CD38 antigen.

An ecto-enzyme of NAD glycohydrolase (NADase) induced by retinoic acid in HL-60 cells is attributed to the molecule of CD38 antigen [Kontani, K., Nishina, H., Ohoka, Y., Takahashi, K., and Katada, T. (1993) J. Biol. Chem. 268, 16895-16898]. CD38 antigen has an amino acid sequence homologous to Aplysia ADP-ribosyl cyclase which generates cyclic adenosine diphosphoribose (cADPR) and nicotinamide (NA) from beta-NAD+. On the basis of this sequence homology, we compared enzyme properties between CD38 NADase expressed as a fusion protein in Escherichia coli and ADP-ribosyl cyclase purified from the ovotestis of Aplysia kurodai. 1) beta-NAD+ analogs, nicotinamide 1, N6-ethenoadenine dinucleotide, and nicotinamide hypoxanthine dinucleotide, did not serve as good substrates for the ADP-ribosyl cyclase, suggesting that the intact adenine ring of beta-NAD+ was required for the cyclase-catalyzed reaction. On the other hand, CD38 NADase utilized the NAD analogs to form ADP-ribose and NA. 2) Kinetic analyses of the ADP-ribosyl cyclase reaction revealed that NA was first released from the substrate (beta-NAD+)-enzyme complex, followed by the release of another product, cADPR, which was capable of interacting with the free enzyme. 3) The enzyme reaction catalyzed by the ADP-ribosyl cyclase was fully reversible; beta-NAD+ could be formed from cADPR and NA with a velocity similar to that observed in the degradation of beta-NAD+. However, CD38 NADase did not catalyze the reverse reaction to form beta-NAD+ from ADP-ribopase and NA. 4) The CD38 NADase activity was, but the ADP-ribosyl cyclase activity was not, inhibited by dithiothreitol.(ABSTRACT TRUNCATED AT 250 WORDS)

ADP-ribosyl Cyclase

Effects of serine protease and deoxyribonuclease on intravascular tumor cell arrest in rat blood-borne lung metastasis.

Effects of alpha-chymotrypsin and deoxyribonuclease I (DNase I) on the early phase of blood-borne lung metastasis in rats were studied using confocal laser scanning microscopy and AH-109A cells labeled either with carboxyfluorescein diacetate succinimyl ester or with 51Cr. Intravenous administration of alpha-chymotrypsin or DNase I appeared to enhance or inhibit, respectively, the tumor cell arrest in lung microvasculature as a reflection of reduction or promotion of the tumor cell clearance in the microvasculature. These results suggest that the effects of alpha-chymotrypsin and DNase I are related to induction of tumor cell aggregation and disaggregation in the blood stream, respectively.

Animals

Mutant genes of gramicidin S synthetase 1 defective in phenylalanine racemization have the same sequence as the wild gene.

Mutant grs1 genes were cloned and sequenced from the Bacillus brevis Nagano BI-4, C-3, E-1, and E-2 strains, which produce defective gramicidin S synthetase 1 (GS1), lacking racemase activity. Surprisingly, these mutant genes had entirely the same sequence as that of the wild type gene. These mutant strains also produce defective gramicidin S synthetase 2 (GS2), lacking 4'-phosphopantetheine, a prosthetic group of this enzyme. The participation of this group in phenylalanine racemization is suggested.

Amino Acid Isomerases

Topology of cytochrome b558 in neutrophil membrane analyzed by anti-peptide antibodies and proteolysis.

Cytochrome b558 is an important constituent of the superoxide-generating system in neutrophils and B lymphocytes. In this paper, the topology of the cytochrome in human neutrophil membrane was studied using antibodies raised in rabbits against synthetic peptides corresponding to various regions in the large and small subunits of the cytochrome. The antibodies recognized the cytochrome subunits in immunoblots and the cytochrome in situ. An antibody raised against residues 150-172 in the large subunit (anti-L123) bound to intact neutrophils, indicating that this region is exposed to the outside of the cells. In contrast, antibodies raised against any of the carboxyl-terminal regions of the large and small subunits (anti-LC and anti-SC, respectively) or the amino-terminal region of the small subunit (anti-SN), bound to neutrophils only after the cells were made permeable by freezing and thawing. The region close to the carboxyl terminus of the large subunit was digested by extracellularly added papain and, as a result, an 18-kDa carboxyl-terminal fragment was detected. Thus the carboxyl-terminal region of the large subunit is cytoplasmic and/or buried in the membrane, and the region around residues 369-398 is exposed on the cell surface. In contrast to the large subunits, the small subunit in neutrophils was resistant to any of the proteinases tested, although the subunit in membrane or Triton-solubilized preparation was digestible with papain. These results indicate that the large subunit of cytochrome b558 is a transmembrane protein with at least two regions exposed on the cell surface and that the carboxyl terminus of this subunit and both termini of the small subunit are exposed to the cytoplasmic side.

Amino Acid Sequence

Small cell lung cancer with extensive cutaneous and gastric metastases.

A case of a 60-year-old Japanese woman with small cell lung cancer involving skin and stomach is reported. She was diagnosed as primary small cell lung cancer accompanied by extensive cutaneous metastases. Three months after the last chemotherapy, she complained of nausea and vomiting. Brain CT scan showed no evidence of central nervous system involvement. Upper gastrointestinal study and upper gastrointestinal fiberscopy revealed multiple metastatic gastric tumors. Skin and stomach are uncommon metastatic sites for any malignancy. Furthermore, only a few cases with gastric metastasis could be diagnosed during their lifetime.

Carcinoma, Small Cell

Transrectal ultrasound-guided interstitial radiation therapy for localized prostate cancer.

The use of interstitial implants for the treatment of low-stage prostate cancer using transrectal ultrasound guidance is evaluated in 80 patients. This outpatient procedure involves the placement of needles through a template and into the prostate. Ultrasound guidance is used to place the needles into a preselected location. The needles are loaded with a radioactive source. In this study Palladium-103 was utilized. This technique allows accurate and complete seeding of the prostate. There was a 50 percent or greater decrease in prostate size in all of the patients who were implanted. Prostate-specific antigen (PSA) levels became normal or decreased by more than 50 percent in 97 percent of the patients. Most patients experienced urethral irritative symptoms which lasted up to five months, but none of the patients experienced rectal symptoms lasting longer than a month. The mean follow-up is 11.8 months which is too brief to ascertain the effectiveness of this therapy. The method appears to be safe and may represent an alternative to external beam irradiation.

Antigens, Neoplasm

The nucleotide sequence for a proline-activating domain of gramicidin S synthetase 2 gene from Bacillus brevis.

A fragment encoding proline-activating domain (grs 2-pro) of gramicidin S synthetase 2 (GS 2) was found in an 8.1-kilobase pairs (kb) DNA fragment of Bacillus brevis Nagano, which contained the full length of GS 1 gene (grs 1). The clones designated GS719 and GS708, which expressed gramicidin S synthetase 1, were elucidated to express immunoreactive proteins to GS 2 antibodies with approximate molecular weights of 115,000, 105,000 (GS719), and 110,000 (GS708). The partial purification of the gene products of these clones was carried out using DEAE-Sepharose CL-6B column chromatography. The immunoreactive proteins to GS 2 antibodies were separated from gramicidin S synthetase 1 protein and had specific proline-dependent ATP-32PPi exchange activity. The nucleotide sequence for the proline-activating domain in the 8.1-kb insert was determined. This fragment was 2,879 base pairs long, and encoded 959 amino acids. The calculated molecular weight of 111,671 was consistent with the apparent molecular weight of 115,000 found in SDS-PAGE of the immunoreactive products to GS 2 antibodies. The open reading frame for this protein followed grs 1 gene, though two were separated by a 73-base pair noncoding sequence, and remained open to the end.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Isomerases

An investigation of an infrared ray electronic endoscope with a laser diode light source.

An infrared ray electronic video-endoscope with a laser diode light source was used to obtain information on the submucosal area. As contrast medium we employed indocyanine green, which has a light absorption peak in the infrared range. The wavelength of the laser diode was set to 810 nm as suggested by the results of the spectrophotometric study on the reflected light at fingertips after injection of ICG (3 mg/kg). Using this system we inspected the stomach under visible light and then continuously under infrared light before and after intravenous injection of ICG (2 mg/kg). A dendritic figure with fine ramifications emerged in the early stage after intravenous injection of indocyanine green, which was not seen under visible light. These features of the figure visualized by the infrared endoscope suggest that they have a close relationship with blood vessels and blood circulation beneath the mucosal surface. This infrared electronic endoscope system provides submucosal information more readily and at a lower cost than ever before, and also the infrared lighting method of this system could be applied to almost all available electronic endoscopes by modulating infrared cutting filters, so it could become a valuable clinical tool.

Electronics, Medical

[Effect of maternally derived antibody levels on antibody responses to canine parvovirus, canine distemper virus and infectious canine hepatitis virus after vaccinations in beagle puppies].

Antibody titers against canine parvovirus (CPV), canine distemper virus (CDV) and infectious canine hepatitis virus (ICHV) in serum were measured in 6 beagle dams and their 38 puppies bred in our colony, in order to clarify the effects of maternally derived antibodies to antibody responses against the viruses after vaccinations in puppies. Correlation coefficient on antibody titers between puppies and dams were CPV: r = 0. 7935, CDV: r = 0.8194 and ICHV: r = 0.8105. Mean maternal antibody positive rates in 7-day-old puppies from their dams were CPV: 67%, CDV: 46% and ICHV: 45%. Mean half-lives of the maternal antibodies in puppies were estimated to be CPV: 13.5 days, CDV: 15.1 days and ICHV: 15.4 days. The antibody response against CPV vaccination in puppies was mainly observed in dogs being titers of less 1:5 and positivity was 39% (15/38 puppies) after 1st vaccination at 42 days after birth, and 82% (31/38 puppies) after 2nd vaccination at 70 days. That against CDV vaccination (at 56 days after birth) was seen highly in dogs being titers of less 1:10 and positivity was 53% (20/38). Also that against ICHV vaccination (at 56 days after birth) was seen frequently in dogs being titers of less 20 holds and the rate was 87% (33/38). From these results, it was estimated that the age when high antibody response against each vaccination could be expected in puppies might be CPV: between 40 and 69 days, CDV: between 32 and 92 days and ICHV: between 31 and 52 days, respectively.

Age Factors

[Chronic gastritic changes induced in rats after pyloromyotomy and per os administration of acetic acid-glycerin].

We have experimentally studied whether or not repeated episodes of acute gastric mucosal lesions can induce chronic gastritic changes. Rats received pyloromyotomy and repeated per os administration of acetic acid-glycerin. Gastric mucosal lesions thus induced were studied histopathologically in cardiac glandular areas using morphometric analysis. It was found that pyloromyotomy induced fibrosis between the bottom of the gland and muscularis mucosa. On the other hand, the per os administration of acetic acid-glycerin induced hyperplastic change of mucous neck cells. They were increased in number and in size. Distribution of mucous neck cells were distended vertically to the mucosal surface and towards the bottom of the gland. Greater the amount of acetic acid-glycerin administration, stronger the changes of mucous neck cells, i.e., the distended zone of mucous neck cells spread more and more from the cardiac area. Parietal and chief cells were decreased in number in the course of the mucous neck cell hyperplasia. Branching of glands was occasionally seen in rats receiving greater amount of acetic acid-glycerin. It was concluded that there are basic structural similarity between the gastric mucosal changes induced by repeated episodes of acute gastric mucosal lesions and the change seen in human chronic gastritis. Thus acute gastric mucosal lesions can be one of the cause of chronic gastritis.

Acetates

[Cell proliferation in a triple gastric cancer--a study using the isolated organ perfusion system].

So as to study the cell proliferation in a case with triple gastric cancers, we labeled an operated whole stomach with bromodeoxyuridine (BrdU) by the isolated organ perfusion system using artificial blood. Anti-BrdU immunohistochemical staining of sections from every part of the stomach was performed, and 3 separated cancerous lesions were all diagnosed as well differentiated adenocarcinomas. The labeling index (L.I.) of the cancerous lesions were found to differ from part to part. However, the L.I. tended to be higher in the superficial mucosal areas and in the cell-dense areas of the submucosa. In the almost normal fundic glands, the labeled zone was localized in the neck of the glands. The more metaplastic gastritis was advanced, the more the labeled zone shifted irregularly toward the bottom of the glands. Whenever the interstinal metaplasia was completed, the labeled zone was localized in the bottom of the glands. This method was found useful in studying the cell proliferation in the stomach without causing any harm to the human body.

Adenocarcinoma

[A ten-year observation of malignant changes in a hyperplastic polyp of the stomach--a cellular kinetic study using bromodeoxyuridine (BrdU)].

Reported is the case of a 64 year old man with a hyperplastic polyp of the stomach that underwent malignant changes. The polyp had been followed up both endoscopically and histologically for 10 years. As it proved difficult to diagnose the malignant changes of the polyp through its endoscopic appearance, a part of polypectomized materials was studied by in vitro labeling of the S-phase cells with BrdU. Although the background hyperplastic mucosa somewhat preserved an irregular generative cell zone, the zonal distribution of the labeled cells were completely disturbed in the areas that underwent a malignant transformation, and labeled cells were often seen in the surface areas of the cancerous glands. It is important to study malignant changes in the hyperplastic polyps of the stomach from a cellular kinetic aspect.

Bromodeoxyuridine

Epidemiologic studies on schistosomiasis japonica in a newly found habitat of Oncomelania snails in Japan.

Epidemiologic studies' on schistosomiasis japonica were carried out in the Obitsu River basin of central Boso Peninsula, a newly found habitat of Oncomelania snails in Japan. Forty-five (4.1%) of 1,102 schoolchildren and 64 (10.4%) of 618 junior high school students were positive for the skin test with veronal buffered saline-extracted Schistosoma japonicum antigen. None was positive by the enzyme-linked immunosorbent assay, complement fixation test, double diffusion test in agar, and circumoval precipitin test. About one fifth of 356 adult inhabitants showed positive reactions in the enzyme-linked immunosorbent assay and complement fixation test as well as in the skin test, although only 25 individuals (7%) were positive for the circumoval precipitin test. All of 70 individuals who had shown positive reactions in at least two of the immunological tests were negative for a series of 5 stool examinations with the AMS III method. Release of S.japonicum cercariae was not observed in 15,000 O. hupensis nosophora collected in this area. Repeated examinations by dipping mice into irrigation ditches were negative. These observations, with the fact that all of 15 patients reported from nearby hospitals to have schistosome eggs in their tissues were over 45 years old, suggest that schistosomiasis has become a disease of the past in this area.

Adolescent