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Biomedical subjects

T Beppu

Publications and source records attributed to T Beppu.

At least 19 recordsLinked to original sources

Apatite-forming ability of alginate fibers treated with calcium hydroxide solution.

Calcium alginate fibers were prepared by extruding an aqueous sodium alginate solution into an aqueous calcium chloride solution. The fibers were treated with a saturated aqueous calcium hydroxide solution for various periods and their apatite-forming ability was examined in a simulated body fluid (SBF). The calcium alginate fibers were treated with the aqueous calcium hydroxide solution for periods longer than five days formed apatite on their surfaces in SBF, and their apatite-forming ability improved with increasing calcium hydroxide treatment time. The amount of calcium ions released from the fibers also increased with increasing calcium hydroxide treatment time, resulting in acceleration of nucleation and growth of apatite on the fiber surfaces. The resultant apatite-alginate fiber composite is expected to be useful as a flexible bioactive bone-repairing material.

Alginates↗

Effective extracellular trehalose production by Cellulosimicrobium cellulans.

A bacterium isolated from a petal of Casa Blanca Lily (ST26 strain) produced a marked amount of extracellular trehalose (alpha- d-glucopyranosyl-[1,1]-alpha- d-glucopyranose) in culture medium containing glucose. 16S rDNA-based phylogeny showed that ST26 belongs to, or is related to, Cellulosimicrobium cellulans, a close relative of Cellulomonas spp. Various Cellulomonas strains obtained from culture collections also showed extracellular trehalose productivity, suggesting that trehalose production is a common property of this bacterial genus. ST26 accumulated trehalose in medium supplied with glucose but not with sucrose, glycerol or maltose. Effective extracellular trehalose production by ST26 was achieved by supplying 0.5-1% ammonium sulfate and 0.5-1% CaCO(3). The addition of CaCO(3) adjusted the pH of the culture to around 5.0. The optimized culture conditions yielded trehalose from glucose at a conversion rate of 61%. The addition of ammonium sulfate greatly reduced the dry cell weight of ST26 and intracellular content of trehalose, which suggests that the addition of ammonium sulfate makes ST26 cells leak trehalose into the medium. ST26 effectively propagated in minimal medium containing trehalose as a sole carbon source, which suggests that trehalose serves as a carbohydrate reserve of this organism.

Ammonium Sulfate↗

Antiapoptotic effect of haem oxygenase-1 induced by nitric oxide in experimental solid tumour.

Induction of haem oxygenase-1 (HO-1) may provide an important protective effect for cells against oxidative stress. Here, we investigated the mechanism of cytoprotection of HO-1 in solid tumour with a focus on the antiapoptotic activity of HO-1. Treatment of rat hepatoma AH136B cells with the HO inhibitor zinc protoporphyrin IX (ZnPP IX) or tin protoporphyrin IX resulted in extensive apoptotic changes of tumour cells both in vivo and in vitro. Caspase-3 activity of the ZnPP IX-treated hepatoma cells increased significantly. Moreover, ZnPP IX-induced apoptosis was completely inhibited by simultaneous incubation with a specific caspase-3 inhibitor and was partially abrogated by bilirubin, a reaction product of HO. In vivo ZnPP IX treatment did not affect nitric oxide (NO) production and tumour blood flow. Western blot analyses showed that HO-1 expression in AH136B cells was strongly upregulated by NO donors, for example, S-nitroso-N-acetyl penicillamine and propylamine NONOate in vitro; conversely, it was remarkably reduced in vivo by pharmacological blockade of NOS. We conclude that HO-1 may function in antiapoptotic defense of the tumour, and thus it may have important protective and beneficial effects for tumour cells against oxidative stress induced by NO, which is produced in excess during solid tumour growth in vivo.

Animals↗

Apatite-forming ability of carboxyl group-containing polymer gels in a simulated body fluid.

Carboxymethylated chitin, gellan gum, and curdlan gels were soaked in a simulated body fluid (SBF) having ion concentrations nearly equal to those of human blood plasma. Some of the gels had been soaked in a saturated Ca(OH)(2) solution, while others had not. The carboxymethylated chitin and gellan gum gels have carboxyl groups, while the curdlan gel has hydroxyl groups. None of the gels formed apatite on their surfaces in the SBF when they had not been subjected to the Ca(OH)(2) treatment, whereas the carboxymethylated chitin and gellan gum gels formed apatite on their surfaces when they had been subjected to the Ca(OH)(2) treatment. The curdlan gel did not form an apatite deposit even after the Ca(OH)(2) treatment. Apatite formation on the carboxymethylated chitin and gellan gum gels was attributed to the catalytic effect of their carboxyl groups for apatite nucleation, and acceleration of apatite nucleation from released Ca(2+) ions. This result provides a guiding principle for obtaining apatite-organic polymer fiber composites. This composite is expected to have an analogous structure to that of natural bone.

Apatites↗

Ixodid tick species recovered from domestic dogs in Japan.

The species of ixodid ticks (Acari: Ixodidae) recovered from domestic dogs in Japan between September to November 2000 and April to June 2001 were identified. A total of 4122 ticks, including 1624 larvae, 1200 nymphs, 1016 females and 282 males were removed from 1221 dogs during these periods. Haemaphysalis longicornis (Neumann) was the most frequently found (40.3% of dogs), followed by H. flava (Neumann) (16.1% of dogs), Rhipicephalus sanguineus (Latreille) (4.8% of dogs) and Ixodes ovatus (Neumann) (4.1% of dogs). Small numbers of H. hystricis (Supino), H. campanulata (Warburton), H. japonica (Warburton), H. ias (Nakamura and Yajima), I. persulcatus (Schulze), I. nipponensis (Kitaoka and Saito) and Amblyomma testudinarium (Koch) were also recovered. In the spring sample, a total of 1408 ticks (78 larvae, 411 nymphs, 792 adult females and 127 adult males) were recovered from 570 dogs. The autumn sample included a larger proportion of larval stage and fewer adult ticks (1546 larvae, 789 nymphs, 224 adult females and 155 adult males). Haemaphysalis longicornis, H. flava and I. ovatus showed a wide geographical distribution from northern to southern Japan, whereas R. sanguineus were mainly distributed in the subtropical Okinawa prefecture with a few exceptions. Dogs in rural areas more frequently carried H. longicornis, H. flava and I. ovatus than dogs in urban or suburban areas, whereas R. sanguineus was more associated with the dogs in urban/suburban areas. Exposure to a garden was significantly associated with R. sanguineus and exposure to woodland was significantly associated with H. flava and I. ovatus. This is the first systematic survey of canine ticks in Japan.

Animals↗

Development of a membrane dialysis bioreactor and its application to a large-scale culture of a symbiotic bacterium, Symbiobacterium thermophilum.

A simple membrane dialysis bioreactor was developed for a large-scale axenic culture of Symbiobacterium thermophilum, a symbiotic thermophile that requires co-cultivation with an associating thermophilic Bacillus strain S for normal growth. The bioreactor consisted of an outer- and an inner-coaxial cylindrical compartment bordered across a dialyzing membrane, which enabled a 1 l-scale dialysis culture with exchange of low molecular metabolites between the two compartments to be performed. Using the bioreactor, growth characteristics of S. thermophilum and Bacillus strain S were assessed under two medium conditions. The growth of S. thermophilum was measured by quantitative PCR because the bacterium formed no visible colonies and gave abnormally low turbidity. In medium containing 2% tryptone peptone, S. thermophilum proliferated up to 4x10(7) cells/ml, and strict dependence on the co-culture with Bacillus strain S was observed. On the other hand, medium containing 0.5% yeast extract not only facilitated the growth of S. thermophilum in the co-culture (6x10(7) cells/ml), but also allowed limited pure growth independent of Bacillus strain S (1x10(7) cells/ml), implying that some component of yeast extract can partially replace the growth requirement of S. thermophilum supplied by Bacillus strain S. Both the oxidative redox potential values and the cell morphology in the independently growing culture suggested the occurrence of marked unbalanced growth possibly caused by significant metabolic changes. The bioreactor is applicable to the analyses of culturing characteristics in symbiotic systems between free-living microorganisms.

Bioreactors↗

Brain PET and technetium-99m-ECD SPECT imaging in Lhermitte-Duclos disease.

Two patients with Lhermitte-Duclos disease were evaluated by brain positron emission tomography (PET) and technetium-99m-ethyl cysteinate dimer (99mTc-ECD) single-photon emission computed tomography (SPECT). In the lesions in both patients, hyperperfusion was detected on cerebral blood flow images obtained by PET, and hyperactivity by standard 99mTc-ECD SPECT. Dynamic 99mTc-ECD SPECT images demonstrated a plateau of activity in each lesion. These findings suggest that lesions in Lhermitte-Duclos disease have a retention mechanism for 99mTc-ECD equivalent to that of normal neural tissue.

Adolescent↗

Double cancers in the common bile duct: molecular genetic findings with an analysis of LOH.

We report a 69-year-old man with double cancers in the common bile duct. One cancer was located between the superior and middle parts of the bile duct, while the other cancer was in the inferior part of the bile duct. Pylorus-preserving pancreatoduodenectomy was performed. There was no communication between the two cancers in either the mucosal layer or the subepithelial layer. On pathological examination, the upper cancer was diagnosed as poorly differentiated adenocarcinoma, while the lower one was found to be moderately differentiated adenocarcinoma. We analyzed loss of heterozygosity (LOH), using microsatellite markers on five chromosomal arms, in both the upper and the lower cancers. Both cancers showed common regions of LOH at 5q, 6q, 9p, 17p, and 18q, whereas the upper cancer showed one additional region of LOH at 8p, thus suggesting progression, due to the acquisition of the additional LOH, in the upper cancer. No LOH was observed in the region between the two cancers. The presence of one additional LOH in the upper cancer suggests that the upper cancer was a metastasis of the lower one.

Adenocarcinoma↗

A new procedure of percutaneous microwave coagulation therapy under artificial hydrothorax for patients with liver tumors in the hepatic dome.

Percutaneous microwave coagulation therapy (PMCT) has been widely used as an effective minimal invasive therapy for small liver tumors. The occurrence of a sonographic masked space due to the presence of the lung, however, has become a major obstacle to visualizing the whole tumor in the hepatic dome. To facilitate the use of PMCT for liver tumors in the hepatic dome, we developed PMCT in combination with the artificial hydrothorax method (percutaneous transdiaphragmatic MCT: PTD-MCT). Our new approach for PMCT to the hepatic tumors located in Couinaud's segments VIII or VII just under the diaphragm resulted in a successful treatment. The separation of the lung from the diaphragm by the infusion of saline into the pleural cavity enabled us not only to visualize the whole tumor in the hepatic dome to accurately target the tumor, but also helped us to avoid injuring the lung. PTD-MCT is therefore strongly recommended for the treatment of liver tumors in the hepatic dome.

Aged↗

Replacements of amino acid residues at subsites and their effects on the catalytic properties of Rhizomucor pusillus pepsin, an aspartic proteinase from Rhizomucor pusillus.

Site-directed mutagenesis was carried out to investigate the functional roles of amino acid residues of Rhizomucor pusillus pepsin (RMPP) in substrate-binding and catalysis. Mutations of two amino acid residues, E13 in the S3 subsite and N219 in the S3/S4 subsites, caused marked changes in kinetic parameters for two substrate peptides with different sequences. Further site-directed mutagenesis at E13 suggested that E13 plays a critical role in forming the correct hydrogen bond network around the active center. In the crystal structure of Rhizomucor miehei pepsin (RMMP), which is an aspartic proteinase produced by Rhizomucor miehei and shows 81% amino acid identity to RMPP, the Oepsilon atom of N219 forms a hydrogen bond with the N-H of isovaline in pepstatin A, a statine-type inhibitor, at the P3 position, suggesting that the loss of the hydrogen bond causes an unfavorable arrangement of the P3 residue. Among the mutants constructed, the E13A mutant showed a 5-fold increase in the ratio of clotting versus proteolytic activity without significant loss of clotting activity. This mutant may present a promising candidate for a useful milk coagulant.

Amino Acid Substitution↗

Percutaneous embolization of the distal pancreatic duct to treat intractable pancreatic juice fistula.

Pseudocysts and post-necrotic collections of the pancreas are sometimes treated by percutaneous drainage. In cases of post-necrotic collection, intractable pancreatic juice fistula is often formed by disruption of the main pancreatic duct in the necrotized region. We radically treated intractable pancreatic juice fistulae by selective cannulation into the distal pancreatic duct via the route for percutaneous drainage of post-necrotic collections to extinguish the exocrine function of the caudal pancreas. We performed this procedure in two patients in whom the major pancreatic duct was damaged at the body of the pancreas, which was extensively necrotic. Although mild symptoms of acute pancreatitis appeared in both patients after the first procedure, they recovered without severe side effects. Neither recurrence of pancreatic juice fistulae nor reduction of the glucose tolerance was caused by removing the exocrine function of the caudal pancreas in either patient 32 and 24 months after treatment, respectively. This method is an effective treatment modality with which to treat intractable pancreatic juice fistulae with damage of the main pancreatic duct.

Adult↗

Distribution and diversity of symbiotic thermophiles, Symbiobacterium thermophilum and related bacteria, in natural environments.

Symbiobacterium thermophilum is a tryptophanase-positive thermophile which shows normal growth only in coculture with its supporting bacteria. Analysis of the 16S rRNA gene (rDNA) indicated that the bacterium belongs to a novel phylogenetic branch at the outermost position of the gram-positive bacterial group without clustering to any other known genus. Here we describe the distribution and diversity of S. thermophilum and related bacteria in the environment. Thermostable tryptophanase activity and amplification of the specific 16S rDNA fragment were effectively employed to detect the presence of Symbiobacterium. Enrichment with kanamycin raised detection sensitivity. Mixed cultures of thermophiles containing Symbiobacterium species were frequently obtained from compost, soil, animal feces, and contents in the intestinal tracts, as well as feeds. Phylogenetic analysis and denaturing gradient gel electrophoresis of the specific 16S rDNA amplicons revealed a diversity of this group of bacteria in the environment.

Animals↗

Radicicol binding to Swo1/Hsp90 and inhibition of growth of specific temperature-sensitive cell cycle mutants of fission yeast.

A panel screening using cdc mutants of Schizosaccharomyces pombe identified radicicol as a potent growth inhibitor of certain mutants at the permissive temperature. The strains sensitive to radicicol were cdc7, cdc11, and cdc14, all of which are defective in early septum formation. Cytokinesis but not nuclear division of these mutants was inhibited by radicicol, but that of cells with the wild-type background was not. A biologically active derivative of radicicol with a biotin moiety at the C-11 position bound Swo1, an Hsp90 homologue in S. pombe. Increased Swo1 expression partially suppressed radicicol sensitivity of cdc14 and almost completely rescued morphological abnormalities in cdc14 and cdc7 cells induced by radicicol at the permissive temperature. On the other hand, the increased Swo1 expression did not restore septum formation at the nonpermissive temperature. These results suggest that Swo1, as a molecular chaperone, plays a role in stabilizing these temperature-sensitive proteins at the permissive temperature or in activating the cytokinesis signaling cascade.

Antifungal Agents↗

Characterization of mutants defective in melanogenesis and a gene for tyrosinase of Streptomyces griseus.

Nitrosoguanidine-induced melanin-negative mutants of Streptomyces griseus were classified according to their ability to produce streptomycin and A-factor, and to form aerial mycelium. A large proportion of the mutants showed deficiency in either antibiotic production and morphological development or both, suggesting close regulatory correlation between melanogenesis and morphological and physiological differentiation. The tyrosinase-encoding mel operon of S. griseus was cloned and examined for its role in melanogenesis of this organism. As in other Streptomyces homologues, the operon consisted of two open reading frames, melC1 encoding the putative cofactor and melC2 encoding the tyrosinase. Regardless of the distinct sequence similarity, introduction of the operon on plasmids failed to confer melanin production in the melanin-negative mutants, and the disruption of melC2 barely affected the melanin productivity, which indicated the presence of another enzyme involved in the melanogenesis in S. griseus. On the other hand, mel on a high-copy-number plasmid caused precocious aerial mycelium formation in Streptomyces lividans TK21 suggesting a stimulatory role of tyrosinase in morphological development.

4-Butyrolactone↗

[A case report of aspergillus granuloma in the cavernous sinus].

We encountered a case of aspergillus granuloma in the cavernous sinus originating in the left sphenoid sinus. The patient was a 56-year-old woman who complained of visual loss and cavernous sinus syndrome on the left side. Hematological data were within normal limits. Bone CT showed evidence of bone destruction in the roof of the left sphenoid sinus. MR images with gadolinium demonstrated an enhanced heterogeneous mass extending from the left sphenoid sinus to the cavernous sinus and the orbital apex. Angiography showed occlusion of the left internal carotid artery. Dexamethasone was administered intravenously every day, because the patient was diagnosed as having Tolosa-Hunt syndrome (THS). However, dexamethasone was not effective. We carried out a biopsy of the mass from the sphenoid sinus, and findings showed aspergillosis. On the 19th day after initiation of fluconazole treatment, the patient lapsed into unconsciousness and presented left hemiparesis. T2-weighted MR images showed high intensity areas in the pons and the thalamus. The patient died two days later. An autopsy revealed edema and many infarctions throughout the brain. The cavernous sinus had been replaced completely with aspergillus granuloma. Furthermore, numerous hyphae of aspergillus had adhered to the inside of the basilar artery, and caused occlusions of arteries around the brain. This case indicated the difficulty of diagnosis of THS and the hazard of over-application of steroids. In this paper, we describe the clinical and pathological behavior of aspergillosis in the cavernous sinus.

Aspergillosis↗

[Efficacy of local ablation therapy for liver metastasis from colorectal cancer--radiofrequency ablation and microwave coagulation therapy].

We evaluated the efficacy of local ablation therapy in 40 patients with liver metastases from colorectal cancer. Radiofrequency ablation (RFA) and/or microwave coagulation therapy (MCT) were used. Ablation therapies were performed in percutaneous, endoscopic, and operative procedures. The regional recurrence rate at the therapeutic sites was 15% (median follow-up period of 2.5 years). The average surgical margin in the operative ablation group was 11 mm. The cumulative 5-year survival rates were 37% in the local ablation, 41% in the hepatic resection, and 5% in the regional chemotherapy groups. Major complications occurred in only two patients (one biliary fistula and one liver abscess). Together these observations indicate that local ablation therapy is a radical and safe locoregional therapy that provides adequate local control and contributes to long survival.

Catheter Ablation↗