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Biomedical subjects

S Yoshioka

Publications and source records attributed to S Yoshioka.

At least 19 recordsLinked to original sources

Purification and properties of Aspergillus niger beta-glucosidase.

Beta-glucosidase was purified from a crude cellulase preparation from Aspergillus niger by affinity chromatography on a methacrylamide-N-methylene-bis-methacrylamide copolymer bearing cellobiamine. The purified enzyme was a dimer with an isoelectric point of 4.0. The molecular mass of the enzyme was estimated to be 240 kDa by gel-permeation chromatography. The enzyme hydrolyzed specifically beta-glucosidic bonds and catalyzed transglucosylation of the beta-glucosyl group of cellobiose to yield 4-O-beta-gentiobiosylglucose in the presence of organic solvents or under neutral conditions.

Aspergillus niger

Effect of water mobility on drug hydrolysis rates in gelatin gels.

The stability of drugs incorporated in gelatin gels was studied, with a focus on the water mobility in the gels. Trichlormethiazide hydrolysis and kanamycin-catalyzed flomoxef hydrolysis in gelatin gels were chosen as models for apparent first-order and second-order hydrolysis, respectively. The mobility of water in gelatin gels was determined by NMR, ESR, and dielectric relaxation spectroscopies. The amount of bound water in the gels was determined from dielectric relaxation spectra. Spin-lattice relaxation time of water determined by 17O NMR and rotational correlation time of an ESR probe determined by an ESR probing method were useful in determining the microviscosity of the gels. The hydrolysis rate of trichlormethiazide in the gels was found to depend on the amount of free water available for the reaction, while that of flomoxef depended on the microviscosity of the gels, which reflected the mobility of water molecules. Thus the dependence of hydrolysis rates on the water mobility was influenced by the hydrolysis mechanism.

Cephalosporins

Differential diagnosis of idiopathic fibrosis from malignant lymphadenopathy with PET and F-18 fluorodeoxyglucose.

Idiopathic fibrosis and malignant lymphoma were studied with positron emission tomography (PET) using 2-deoxy-2-[F-18]fluoro-D-glucose (F-18DG). Cases of retroperitoneal fibrosis and sclerosing peritonitis showed low F-18DG uptake with the mass/muscle radioactivity (M/M) ratio 2.0 and 1.9, respectively. A case of para-aortic adenopathy from malignant lymphoma showed high F-18DG uptake with the M/M ratio 9.4. F-18DG PET study may be useful for the differential diagnosis of idiopathic fibrosis and malignant lymphadenopathy.

Aged

High serum alkaline phosphatase level of meningioma cell origin: case report and review of the literature.

A 13-year-old boy with an anaplastic meningioma at the site of the jugular foramen had an increased serum level of aklaline phosphatase (ALPase) (liver form) in the serum before surgery. Immediately after excision of the tumor, the serum ALPase level decreased dramatically. Histochemical and immunohistochemical examinations revealed ALPase activity (liver form) in the neoplastic cells. This case would be the third with clinical evidence that the increased level of serum ALPase is of neoplastic cell origin from the meningioma. The implications of ALPase in brain tumors are discussed.

Adolescent

Metabolic fate of [14C]triglyceride-entrapped lactosylceramide-bearing liposomes after intravenous injection into mouse.

We investigated the distribution and fate of liposomes after their intravenous injection into a mouse. Liposomes were composed of dimyristoylphosphatidylcholine, cholesterol and dicetylphosphate (7:2:1, molar ratio) with or without lactosylceramide. They were characterized as small unilamellar vesicles, approximately 100 nm in diameter, using gel-exclusion chromatography on a Sephacryl S-1000, freeze-fracture electron microscope and dynamic light scattering method. Liposomes were very stable in serum as seen by the results of leakage of the entrapped marker and electrophoresis experiments. We demonstrated that liposomes were internalized by way of an endocytotic process via coated vesicles detected in the electron microscope. The increase in liver uptake of lactosylceramide-bearing liposomes was mostly accounted for by enhanced uptake in the parenchymal cells, while uptake by non-parenchymal cells was only slightly increased. This observation supported the notion that a galactose-specific receptor was involved in liver uptake of lactosylceramide liposomes. The lactosylceramide-bearing liposomes were preferentially recovered in the liver and were found to first be predominantly localized in the mitochondria-lysosomal fraction. They were then decomposed by lysosomal enzymes, and the hydrolyzed components were reincorporated into membrane phospholipids in the microsomal fraction. At the same time, a rapid and reversible exchange of phosphatidylcholine between microsomes and mitochondria was demonstrated.

Absorption

Effect of radiation therapy against intracranial hemangiopericytoma.

Seven cases of intracranial hemangiopericytoma were studied retrospectively to investigate the efficacy of radiation therapy. Tumor response evaluated by computed tomography and magnetic resonance imaging was obvious after 20-30 Gy irradiation. The total reduction rate was 80-90% and continued as long as 5-7 months after treatment. In five patients receiving radiation therapy before radical removal, the tumors were easily removed without massive hemorrhage. Histological inspection of specimens after irradiation showed a significant disappearance of tumor cells. Pyknosis frequently occurred in endothelial cells, and proliferating vessels with hyalinoid degeneration were also seen. Reticulin fibers between tumor cells were fewer, split, or absent. Preoperative radiation therapy is useful in the treatment of hemangiopericytoma involving considerable surgical risk. Postoperative radiation therapy should be given even if removal is complete.

Adolescent

[Brain metastases from Wilms' tumor without systemic involvement--a case report and review].

A 3-year-old boy presented with multiple brain metastases 21 months after the resection of stage II Wilms' tumor. Metastasis to other organs was not found. He was treated by total removal of a large metastatic tumor in the left temporal lobe and post-operative radiotherapy and chemotherapy. He has been in complete remission for 20 months after surgery. Cerebral metastasis from Wilms' tumor without systemic metastases is very rare. It is speculated that brain metastases occurred in this patient because most of the anticancer agents used in the primary treatment for Wilms' tumor were not able to cross the blood brain barrier.

Antineoplastic Agents

[Urodynamic study on urinary disturbance after therapy of uterine cancer].

It is well known that urinary disturbance often appears after radical hysterectomy for uterine cancer and is aggravated by additional radiation therapy. The aim of this study is to elucidate the pathogenesis and to establish the treatment method of urinary disturbance after therapy for uterine cancer. Forty-five patients with urinary disturbance and ten normal controls were subjected to this study. Changes in clinical symptoms and findings in the Urodynamic study (UDS) in 12 severe cases were investigated before and after treatment with beta 2-stimulant (Mabuterol HCL). Clinical symptoms in cases treated by radiation therapy alone were rare and mild without any pad exchanges, and appeared 5 years after treatment for uterine cancer. Findings of UDS in these cases were mild low compliance of detrusor at maximum desire to void (Cmdv) and mild low bladder volume at maximum desire to void (Vmdv). In cases treated by radical hysterectomy alone, Cmdv decreased immediatelly after the operation and then maximum urethral closure pressure (cPura) gradually decreased. Concerning the cases treated by radical hysterectomy and radiation, severe low Cmdv and severe low Vmdv appeared 5 years after the treatment for uterine cancer in almost all cases, and low cPura appeared immediately after the operation in half of the cases. Treatment with beta 2-stimulant significantly improved urinary frequency, voided volume and urinary incontinence. In UDS findings, Vmdv and Cmdv were significantly improved by the treatment with beta 2-stimulant. The functional profile length and cPura value were not significantly changed. In uroflowmetry, the maximum flow rate and average flow rate were significantly improved by the treatment with beta 2-stimulant.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic beta-Agonists

Characterization of CS-045, a new oral antidiabetic agent, II. Effects on glycemic control and pancreatic islet structure at a late stage of the diabetic syndrome in C57BL/KsJ-db/db mice.

Antidiabetic effects of CS-045 were evaluated in 5-month-old C57BL/KsJ-db/db mice (db/db). CS-045 administered for 3 weeks to diabetic db/db mice as a 0.2% food admixture improved hyperglycemia (855 +/- 25 v 298 +/- 62 mg/dL, P less than .01) and glucose intolerance, and lowered plasma triglyceride (299.6 +/- 28.7 v 76.3 +/- 20.7 mg/dL, P less than .01) and free fatty acid (FFA) levels (1.16 +/- 0.14 v 0.57 +/- 0.07 mEq/L, P less than .01). Food intake was not changed, while a small but significant increase in body weight was observed in CS-045-treated mice. Plasma insulin levels gradually increased after 5 days of CS-045 treatment, and a nonsignificant increase was observed in plasma insulin levels after 3 weeks (1.85 +/- 0.50 v 4.54 +/- 1.47 mg/mL). In contrast, the plasma glucagon levels decreased after 3 weeks of CS-045 treatment. Histological examination by aldehyde-fucshin staining demonstrated that pancreatic beta cells in CS-045-treated db/db mice were heavily regranulated, whereas most of the beta cells were extensively degranulated in nontreated db/db mice. The heavily regranulated state of beta cells was also compatible with an increase in pancreatic insulin content in CS-045-treated db/db mice. Electron microscopic analysis showed a well-developed endoplasmic reticulum and the accumulation of much amorphous structural material in the intracisternal space of beta cells from CS-045-treated db/db mice, which were suggestive of an increase in insulin synthesis. Moreover, CS-045 treatment decreased exocrine-containing islets, which was associated with the islets' degeneration process. Immunohistochemical staining of islets showed that CS-045 treatment normalized the distribution pattern of endocrine cells in the islets of db/db mice, reflected by a predominantly peripheral location of alpha and delta cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral

Potential radiopharmaceuticals labeled with titanium-45.

The potential of some compounds labeled with cyclotron-produced titanium-45 (45Ti) as radiopharmaceuticals was studied. Properties of colloid formation of 45TiOCl2 or 45TiO-phytate in vivo resulted in the highest radioactivity uptake in the rat liver, followed by the spleen, suggesting potential for imaging the reticuloendothelial system. Three 45TiO-complexes with diethylenetriaminepentaacetic acid, citric acid and human serum albumin showed the highest radioactivity levels in the blood over 6 h. The binding of the 45Ti with plasma transferrin in vitro and in vivo suggested that these compounds can be used for estimating the blood volume. Also, potential as an indicator representing the breakdown of the blood-brain barrier in the rat was demonstrated by autoradiography.

Animals

Inactivation kinetics of enzyme pharmaceuticals in aqueous solution.

The kinetics of enzyme inactivation in aqueous solution of neutral pH were studied for alpha-chymotrypsin, bromelain, and kallikrein. Inactivation of alpha-chymotrypsin and bromelain followed simple first-order kinetics, and the rate constant obtained conformed to the Arrhenius relationship. Kallikrein, however, presented more complicated kinetics of inactivation, which could be described by a kinetic expression combining a reversible and an irreversible pathway. Nonlinear regression analysis suggested that the rate constants conform reasonably well to the Arrhenius relationship. The results suggest that inactivation of enzymes in aqueous solution can be modeled even if the profile is complicated and that the inactivation rates can be predicted based on the relationship between the parameter estimates and temperature.

Bromelains

Changes of anti-HB core antibody in children with positive HB surface antigen.

To elucidate the clinical courses of children chronically infected with HB virus (HBV), RIA values of anti-HBc were surveyed in 88 cases with positive HBs antigen. Among 56 children with positive HBe antigen, 20 had negative, indefinite or low titers of anti-HBc, and 18 (90%) of them had no liver malfunction. Out of 30 cases with abnormal liver function tests, 28 (93%) had high titers of anti-HBc. Follow-up study for a period of over 12 months reveals that serum HBe antigen disappeared in 10 out of the 40 cases who were positive for this antigen. All of the 10 cases had liver malfunction and high levels of anti-HBc. Among 12 children with initially positive anti-HBe, five had high titers of anti-HBc. Out of 13 children who once had high levels of anti-HBc, 3 showed reduction in titers of anti-HBc during the follow-up period in accordance with decrease in activity of hepatitis. Four out of 16 who initially had HBe antigen and low titers of anti-HBc showed high titers of anti-HBc during the observation period, while only one of 33 who had HBeAg and a high titer of anti-HBc went to the low titer group of anti-HBc, though continuously positive for HBe antigen. We presume that high levels of anti-HBc indicate previous or current liver damage due to HBV infection, while low titers of anti-HBc indicate that HBV-derived liver damage has not yet occurred or that a long time has passed since the last episode of liver damage subsided.

Adolescent

Study to establish normal values for portal vein blood flow in children using a duplex ultrasound system.

Portal vein blood flow in healthy children was measured using a duplex ultrasound system consisting of a B mode linear scanner and a single Doppler transducer. Portal blood flow volume (PBFV) correlated with age, height, weight and body surface area. Increases in PBFV with age were greater in boys than girls. A correlation was noted between the diameter of the portal vein and PBFV (r = 0.89). However, the maximum portal blood flow velocity (Vmax) did not correlate with age, height, weight or body surface area. Increases in PBFV in accordance with physical development was thought to depend mainly on widening of the portal vein and not on acceleration of portal blood flow velocity. The formula for calculating PBFV by multivaliate statistical analysis is as follows: PBFV(ml/min) = 30.1 x age(yrs) - 1.07 x height(cm) = 3.31 x weight(kg). Portal vein flowmetry using a duplex ultrasound system may be useful for evaluation of the hepatic circulation in children.

Adolescent

Expression of ClpB, an analog of the ATP-dependent protease regulatory subunit in Escherichia coli, is controlled by a heat shock sigma factor (sigma 32).

Escherichia coli K-12 produces at least two ATP-dependent proteases, Lon (La) and Clp (Ti), the latter consisting of a regulatory subunit (ClpA) and a proteolytic subunit (ClpP). The gene clpB encoding an analog of ClpA had been found at 57 min on the E. coli chromosome. Cloning and examination of novel heat shock promoters led us to identify a major clpB promoter specifically controlled by a heat shock sigma factor, sigma 32 (the rpoH [= htpR] gene product). beta-Galactosidase synthesis from a PclpB-lacZ operon fusion was transiently induced upon temperature shift from 30 to 42 degrees C, and the induction depended on the rpoH function. Chromosomal clpB transcripts also increased upon temperature upshift and were totally absent in the rpoH deletion strain. In the in vitro transcription experiments, the clpB promoter was specifically recognized and transcribed by RNA polymerase-sigma 32. Nucleotide sequencing and determination of mRNA start sites permitted us to identify a major heat shock promoter located upstream of the clpB coding sequence. The results clearly indicate that clpB expression is under direct control of sigma 32. Since ClpP was recently shown to be a sigma 32-dependent heat shock protein, the present finding suggests the possibility that a potential ATP-dependent protease, ClpB-ClpP complex, plays an important role against thermal stress in E. coli.

ATP-Dependent Proteases

Comparison of 1,5-anhydroglucitol, HbA1c, and fructosamine for detection of diabetes mellitus.

To evaluate the use of serum 1,5-anhydroglucitol (AG) levels in screening for diabetes mellitus, we compared the sensitivity and specificity of HbA1c, fructosamine (FA), and AG in 1620 randomly selected subjects in 11 institutions throughout Japan. Most individuals were receiving diet and/or drug therapy for diabetes. Subjects were separated into four groups based on World Health Organization criteria: nondiabetic control subjects, subjects with impaired glucose tolerance (IGT), patients with diabetes, and patients with other disorders without IGT. The overlap of AG values between each group was less than that of HbA1c or FA values. AG levels were significantly correlated with fasting plasma glucose (r = -0.627), HbA1c (r = -0.629), and FA (r = -0.590) levels. If we took 14 micrograms/ml as the normal lower limit, AG level was highly specific (93.1%), and a decreased AG level indicated diabetes mellitus (84.2% sensitivity). According to the selectivity index (sensitivity value times specificity value), AG determinations were superior to both HbA1c and FA measurements for diabetes screening. When combinations of these tests were used, only AG and HbA1c together were slightly better than AG alone. Thus, together with other advantages of AG, e.g., its wide variance with relatively fair glycemic control and the negligible influence of the sampling conditions, AG level has more potential than HbA1c or FA level as a screening criterion for diabetes.

Biomarkers

Basic studies on ursodeoxycholyl-para-aminobenzoic acid for evaluation of intestinal microflora.

A newly synthesized conjugate of ursodeoxycholic acid with para-aminobenzoic acid (PABA) was investigated to determine its suitability for evaluation of enteric bacteria. This compound, PABA-UDCA, was deconjugated by cholylglycine hydrolase to release free PABA, whereas it was completely resistant to deconjugation by pancreatic and intestinal mucosal enzymes. In bacteriologic experiments almost all the microorganisms that split glycocholic acid deconjugated PABA-UDCA. In rat experiments urinary excretions of PABA were measured during 6 h after oral administration of 10 mg PABA-UDCA (PABA-UDCA administration test). Ten control rats excreted 338.5 +/- 13.8 micrograms (mean +/- SE) of PABA; 10 rats with intestinal stagnant loop excreted more (673.6 +/- 70.2 micrograms; P less than 0.01); whereas 10 rats in each of 7 groups pretreated with oral administration of various antibiotics excreted less (P less than 0.001; polymixin B + tinidazole, 14.0 +/- 2.5 micrograms; polymixin B, 224.9 +/- 23.5 micrograms; tinidazole, 42.7 +/- 8.6 micrograms; kanamycin, 50.3 +/- 5.8 micrograms; clindamycin, 57.4 +/- 7.4 micrograms; vancomycin, 70.4 +/- 8.5 micrograms; and paromomycin, 160.4 +/- 16.4 micrograms). This result was reflected by the bacterial mean count of feces. In the PABA-UDCA administration test, after 2 months of feeding with different diets, rats with high-fiber diet (n = 10) excreted less PABA in urine (70.9 +/- 15.9 micrograms; P less than 0.001) than rats on a control diet (n = 10) and a high-protein-high-fat diet (n = 10) (288.9 +/- 34.5 micrograms and 386.7 +/- 61.2 micrograms, respectively). Fecal bacteriologic status was consistently altered. In human volunteers 250 mg PABA-UDCA was tested. Amounts of PABA excreted in urine during 6 h after dosing were 21.11 +/- 2.02 mg in controls (n = 5) and 12.20 +/- 1.01 mg in the group treated with polymixin B plus tinidazole (n = 5; P less than 0.01). No adverse effect was observed. These basic studies indicate that this compound is likely to offer a simple and rapid method for evaluation of the intestinal microorganisms without use of radioisotopes or expensive, special equipment.

4-Aminobenzoic Acid

Stability of vitamin B1 in a commercial multivitamin syrup preparation.

Accelerated stability studies of a commercial vitamin B1 syrup preparation were carried out at 50, 60, 70 and 80 degrees C. The decomposition of vitamin B1 in the preparation followed pseudo-first order reaction kinetics. Apparent decomposition rate constants conformed to the Arrhenius equation. Apparent activation energy and shelf-life at 25 degrees C were estimated to be 24.2 kcal/mol and 3.3 years, respectively.

Dosage Forms