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T Akao

Publications and source records attributed to T Akao.

At least 37 records · Page 2Linked to original sources
PubMed2000

Occurrence of Bacillus thuringiensis in fresh waters of Japan.

Bacillus thuringiensis was recovered at a relatively high frequency from both running and still fresh waters in natural environments of Kyushu, Japan. Of 107 water samples examined, 53 (49.5%) contained this organism. The frequency of B. thuringiensis colonies was 4.4% among 4414 colonies of the Bacillus cereus/B. thuringiensis group. The density of this bacterium in fresh waters averaged 0.45 cfu/ml. Serologically, B. thuringiensis isolates were assigned to 26 H serotypes. Of these, H14/36 (H serovar israelensis/malaysiensis) was the predominant, followed by the serotypes H3abc (kurstaki), H27 (mexicanensis), H3ad (sumiyoshiensis), and H35 (seoulensis). Of 195 isolates, 52 (26.7%) exhibited larvicidal activity against aquatic Diptera; 21 killed Culex pipiens molestus (Culicidae) only, and 31 were active on both the culicine mosquito and the moth-fly, Clogmia albipunctata (Psychodidae). The Diptera-toxic isolates produced spherical or irregularly pointed parasporal inclusions.

Animals↗
PubMed2000

In vitro cytotoxicity of non-cyt inclusion proteins of a Bacillus thuringiensis isolate against human cells, including cancer cells.

A soil isolate designated 90-F-45-14, belonging to Bacillus thuringiensis serovar dakota (H15), was examined for characterization of in vitro cytotoxicity, associated with parasporal inclusion proteins, against human cells. When activated with proteolytic processing, inclusion proteins of the isolate 90-F-45-14 exhibited a moderate cytotoxicity against the human uterus cervix cancer cells (HeLa) with an EC(50) value of 60.8 microg ml(-1), while showing extremely high activities on the human leukaemic T cells (MOLT-4) and the normal T cells with EC(50) values of 0.27 and 0.20 microg ml(-1), respectively. Anti-leukaemic cell activity of the 90-F-45-14 proteins was eight to nine times greater than that of the B. thuringiensis serovar israelensis proteins containing the Cyt1 protein, a broad-spectrum cytolysin. The cytopathy by the 90-F-45-14 proteins was characterized by marked cell-ballooning, while the israelensis proteins induced early breakdown of the cells due to cytolysis. Inclusions of the isolate consisted of five major polypeptides of 170, 103, 73, 40 and 32 kDa. A 100% homology was observed in the sequence of 15 N-terminal amino acids between the proteins of 170 and 103 kDa. There was no N-terminal sequence homology between 90-F-45-14 proteins and the existing Cry/Cyt proteins of B. thuringiensis. Proteolytic processing by proteinase K yielded several proteins with molecular masses ranging from 40 to 28 kDa.

Amino Acid Sequence↗
PubMed2000

The clinical utility of measuring total PSA, PSA density, gamma-seminoprotein and gamma-seminoprotein/total PSA in prostate cancer prediction.

BACKGROUND: To evaluate whether serum total prostate-specific antigen (PSA), PSA density (serum total PSA level divided by prostate volume), gamma-seminoprotein and gamma-seminoprotein/total PSA ratio could predict prostate cancer (PCa) prior to biopsy. METHODS: A total of 316 consecutive patients who had undergone transrectal prostate biopsy and/or transurethral resection were examined. The prostate volume was determined by transrectal ultrasonography (TRUS) and the ability of the above-mentioned four variables to distinguish PCa from benign prostatic hyperplasia (BPH) was evaluated. RESULTS: PCa was detected in 61 cases. Receiver-operating characteristic (ROC) analysis revealed that both the PSA density and serum total PSA were the most useful predictors of PCa among the four variables. For the patients with a serum total PSA level of 4.1-10.0 ng/ml, PSA density was significantly more accurate than total PSA (p < 0.005). An optimum PSA density value of 0.18 was chosen as a cutoff because it showed the highest sum of sensitivity and specificity, 92 and 54%, respectively. Using this PSA density cutoff, the number of biopsies could have been reduced to 57 from 63% when compared with a PSA density of 0.15. CONCLUSIONS: PSA density was significantly more accurate than other variables in predicting PCa. To avoid unnecessary biopsies, the PSA density cutoff value of 0.18 would be recommendable for determining a prostate biopsy for Japanese males with a serum total PSA level of 4.1-10.0 ng/ml.

Aged↗
PubMed2000

Parasporin, a human leukemic cell-recognizing parasporal protein of Bacillus thuringiensis.

An unusual property, human leukemic cell-recognizing activity, associated with parasporal inclusions of a noninsecticidal Bacillus thuringiensis soil isolate was investigated, and a protein (named parasporin in this study) responsible for the activity was cloned. The parasporin, encoded by a gene 2,169 bp long, was a polypeptide of 723 amino acid residues with a predicted molecular weight of 81, 045. The sequence of parasporin contained the five conserved blocks commonly found in B. thuringiensis Cry proteins; however, only very low homologies (<25%) between parasporin and the existing classes of Cry and Cyt proteins were detected. Parasporin exhibited cytocidal activity only when degraded by proteases into smaller molecules of 40 to 60 kDa. Trypsin and proteinase K activated parasporin, while chymotrypsin did not. The activated parasporin showed strong cytocidal activity against human leukemic T cells (MOLT-4) and human uterus cervix cancer cells (HeLa) but not against normal T cells.

Amino Acid Sequence↗
PubMed2000

Baicalin, the predominant flavone glucuronide of scutellariae radix, is absorbed from the rat gastrointestinal tract as the aglycone and restored to its original form.

When baicalin was orally administered to conventional rats, it was detected in their plasma for 24 h after administration, but baicalein, the aglycone of baicalin, was not detected. However, when baicalin was given to germ-free rats, only a small amount of baicalin was detected in their plasma within 2 h after the administration, its AUC0-lim (the area under the concentration-time curve from 0 to last determination time) being 12.0% of that in conventional rats. Subsequently, a considerable amount (55.1 +/- 6.2%) of baicalin was recovered from the gastrointestinal tract even 4 h after administration. When baicalein was orally administered to conventional rats, however, baicalin appeared rapidly in their plasma at an AUC0-lim value similar to that obtained after oral administration of baicalin, despite the absence of baicalein in plasma. When intestinal absorption was evaluated by the rat jejunal loop method, baicalein was absorbed readily, but only traces of baicalin were absorbed. Moreover, in conventional rats a small amount (13.4 +/- 3.1%) of baicalin and an appreciable amount (21.9 +/- 3.4%) of baicalein were recovered from the gastrointestinal tract even 4 h after oral administration of baicalin, but only a small amount (3.93 +/- 1.43%) of baicalein was detected in the intestinal tract 1 h after administration of baicalein. Baicalin was transformed to baicalein readily by the rat gastric and caecal contents. When baicalin was administered orally to conventional rats, an appreciable amount of baicalein was recovered in their gastrointestinal tracts. Moreover, baicalein was efficiently conjugated to baicalin in rat intestinal and hepatic microsomes. These results indicate that baicalin itself is poorly absorbed from the rat gut, but is hydrolysed to baicalein by intestinal bacteria and then restored to its original form from the absorbed baicalein in the body.

Administration, Oral↗
PubMed2000

Effects of glycyrrhizin and glycyrrhetic acid on the growth, glycyrrhizin beta-D-glucuronidase and 3 beta-hydroxysteroid dehydrogenase of human intestinal bacteria.

The peak of glycyrrhizin (GL) beta-D-glucuronidase activity for Ruminococcus sp. PO1-3 and Eubacterium sp. GLH changed to 24 h from 12 h of culture and to 12 h from 48 h, respectively, at almost the same level by the addition of 1.0 mM GL. This enzyme activity was about 20-fold higher in Eubacterium sp. GLH than in Ruminococcus sp. PO1-3. GL beta-D-glucuronidase activity of Ruminococcus sp. PO1-3 with Eubacterium sp. GLH and the intestinal flora showed a maximal peak at 12 h of culture in the presence and absence of 1.0 mM GL. This enzyme activity was about 2.5-fold higher in mixed bacteria than in intestinal flora. 3Beta-hydrosteroid dehydrogenase activity of Ruminococcus sp. PO1-3 and Ruminococcus sp. PO1-3 with Eubacterium sp. GLH was suppressed greater in the presence of GL than without GL. Also, Ruminococcus sp. PO1-3, Eubacterium sp. GLH, and a mixture of both and intestinal flora, metabolized 1.0 mM GL to glycyrrhetic acid (GA) in yields of about 10, 70, 40 and 100%, respectively, with 24 h culture. From the level of GL beta-D-glucuronidase activity, it is considered that the metabolism of GL by intestinal flora is due to both enzymatic and non-enzymatic reactions. Moreover, GA at a concentration of 1.0 mM suppressed growth of Ruminococcus sp. PO1-3, Eubacterium sp. GLH, and the mixture of both and intestinal flora, which metabolized 1.0 mM GA to a negligible amount of 3-oxo-glycyrrhetic acid, indicating the accumulation of unchanged GA. GL beta-D-glucuronidase activity of intestinal flora was enhanced by GA, which stimulated bacteria possessing particular this characteristic.

3-Hydroxysteroid Dehydrogenases↗
PubMed2000

Differences in the metabolism of glycyrrhizin, glycyrrhetic acid and glycyrrhetic acid monoglucuronide by human intestinal flora.

Glycyrrhizin (1.0 mm GL), glycyrrhetic acid (1.0 mm GA) and glycyrrhetic acid monoglucuronide (1.0 mM GAMG), as well as a combination of all components added to medium at the start of growth and at the maximal stage of intestinal flora were cultured for 24 and 12 h, respectively. GL alone enhanced GL beta-D-glucuronidase activity about 2.7- to 6.8-fold and was metabolized to between 55 and almost 100% GA. GAMG alone was metabolized to almost 100% GA by GAMG beta-D-glucuronidase activity. Intestinal flora grown to a maximal stage converted GL to about 15% GA and GAMG to about 13% GA at almost 0 h. GL in combined GL and GA was consumed about 20% at 12 h and about 100% at 24 h under different culture conditions. Metabolite GA and unchanged GA were metabolized to a negligible amount of 3-oxoglycyrrhetic acid, 3alpha-hydroxyglycyrrhetic acid (3alpha-hydroxyGA) or both by 3beta-hydroxysteroid dehydrogenase and 3alpha-hydroxyGA dehydrogenase activities. Combined GL and GAMG consumed about 90% and 100% GAMG at 24 h and 12 h, respectively, regardless of culture conditions, and the consumption of GL was non-existent or negligible. Consumption of combined GL, GA and GAMG was similar to that of both combined GL and GA and combined GL and GAMG. It was found that intestinal flora can metabolize GL alone, but does not readily metabolize GL when present among its metabolites containing GL.

Clostridium↗
PubMed2000

Competition in the metabolism of glycyrrhizin with glycyrrhetic acid mono-glucuronide by mixed Eubacterium sp. GLH and Ruminococcus sp. PO1-3.

Eubacterium sp. GLH possessing glycyrrhizin (GL) and glycyrrhetic acid mono-glucuronide (GAMG) beta-D-glucuronidases, Ruminococcus sp. PO1-3 possessing GL and GAMG beta-D-glucuronidases and 3beta-hydroxysteroid dehydrogenase and these mixed bacteria were cultured in GAM medium with and without GL, GAMG or both. GL added to Eubacterium sp. GLH accelerated the peaks of enhanced GL beta-D-glucuronidase activity and suppressed GAMG beta-D-glucuronidase activity, and GAMG delayed the peaks of the enhanced growth with GL and GAMG beta-D-glucuronidase activities. GL added to Ruminococcus sp. PO1-3 enhanced gradually the growth with GL and GAMG beta-D-glucuronidase activities, and GAMG enhanced slowly GL beta-D-glucuronidase activity and rapidly the growth with GAMG beta-D-glucuronidase activity. The metabolite glycyrrhetic acid (GA) was produced by Eubacterium sp. GLH and Ruminococcus sp. PO1-3 in larger amounts and faster from GAMG than from GL. GL (1.0 mM) and 1.0 mM GAMG added to these mixed bacteria enhanced the growth with GL and GAMG beta-D-glucuronidase activities and were metabolized almost completely to GA in culture of 2 d and 1 d, respectively. It was found that the metabolism of GAMG was faster than that of GL. GL with GAMG added to mixed Eubacterium sp. GLH and Ruminococcus sp. PO1-3 cultured for 0 and 1 d led to a lower level of these enzyme activities and the consumption of GAMG more quickly, not GL. Low GAMG beta-D-glucuronidase had the ability to hydrolyze GAMG well.

Carbohydrate Sequence↗
PubMed2000

Hasty effect on the metabolism of glycyrrhizin by Eubacterium sp. GLH with Ruminococcus sp. PO1-3 and Clostridium innocuum ES24-06 of human intestinal bacteria.

Eubacterium sp. GLH with Ruminococcus sp. PO1-3 and Clostridium innocuum ES24-06 possessing enzymes involved in the metabolism of glycyrrhizin (GL) was cultured in GAM medium with and without 1.0 mM GL or 1.0 mM glycyrrhetic acid (GA). GL (1.0 mM) enhanced 3alpha-hydroxyglycyrrhetinate (3alpha-hydroxyGA) dehydrogenase activity, GA (1.0 mm) suppressed 3alpha-hydroxyGA dehydrogenase activity, GL beta-D-glucuronidase activity and the mixed bacterial growth, and GL and GA showed almost no change in a lower level of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) activity during 5 d of culture. GL (1.0 mM) and GA (1.0 mM) were metabolized to a small amount of GA and a negligible amount of 3-oxo-glycyrrhetic acid (3-oxo-GA) and 3alpha-hydroxyGA, and to a negligible amount of 3-oxo-GA, respectively, by these mixed bacteria. These amounts coincided with those of metabolites produced from 1.0 mM GL and 1.0 mM GA added to these mixed bacteria after 24 h culture. Whole bacteria and sonicated bacteria derived from the collection of these mixed bacteria reached a maximal stage and metabolized GL to a relatively large amount of GA and 3-oxo-GA, and a negligible amount of 3alpha-hydroxyGA and GA to a small amount of 3-oxo-GA and 3alpha-hydroxyGA within 180 min. GL beta-D-glucuronidase with 3beta-HSD and 3alpha-hydroxyGA dehydrogenase partially purified from each bacterium was converted GL to 3alpha-hydroxyGA, producing metabolites of about 60% after 10 min of incubation. These mixed bacteria possessed high enzyme activities could produce the metabolites of GL in under one hour under conditions.

17-Hydroxysteroid Dehydrogenases↗
PubMed2000

Characterization of the anti-cancer-cell parasporal proteins of a Bacillus thuringiensis isolate.

An unusual activity, associated with non-insecticidal and non-haemolytic parasporal inclusion proteins of a Bacillus thuringiensis soil isolate, designated 89-T-26-17, was characterized. The parasporal inclusion of this isolate was bipyramidal, rounded at both ends, containing proteins of 180, 150, 120, 100, and 88 kDa. No homologies with the Cry and Cyt proteins of B. thuringiensis were detected based on N-terminal sequences. Proteolytic processing of the inclusion proteins by proteinase K, trypsin, and chymotrypsin produced a major protein of 64 kDa exhibiting cytocidal activity against human leukaemic T cells and uterus cervix cancer (HeLa) cells. The protease-activated proteins showed no cytotoxicity to normal T cells.

Amino Acid Sequence↗
PubMed1999

Lectin activity of Bacillus thuringiensis parasporal inclusion proteins.

Parasporal inclusion proteins from a total of 151 Bacillus thuringiensis strains, consisting of 139 Japanese isolates and the type strains of 12 H serovars, were screened for haemagglutination (HA) activity against sheep erythrocytes. Of 58 B. thuringiensis strains with HA activity, nine strains exhibited high activity and the remaining 49 strains were moderately active. The strains with high HA activity were derived from phylloplanes and soils of five geographically different localities, and belonged to H serovars kurstaki and other undefined serotype(s). The HA activities in the four selected strains were generated only when alkali-solubilised parasporal inclusion proteins were proteolytically processed. Furthermore, the lectin activity of the four strains was strongly inhibited by preincubation with N-acetylgalactosamine. The lectin-producing B. thuringiensis strains were heterogeneous in other biological activities of parasporal inclusions: insecticidal activity and cytocidal action on human leukaemia T cells.

Animals↗
PubMed1999

Unique activity associated with non-insecticidal Bacillus thuringiensis parasporal inclusions: in vitro cell-killing action on human cancer cells.

Parasporal inclusion proteins from a total of 1744 Bacillus thuringiensis strains, consisting of 1700 Japanese isolates and 44 reference type strains of existing H serovars, were screened for cytocidal activity against human leukaemia T cells and haemolytic activity against sheep erythrocytes. Of 1684 B. thuringiensis strains having no haemolytic activity, 42 exhibited in vitro cytotoxicity against leukaemia T cells. These non-haemolytic but leukaemia cell-toxic strains belonged to several H-serovars including dakota, neoleonensis, shandongiensis, coreanensis and other unidentified serogroups. Purified parasporal inclusions of the three selected strains, designated 84-HS-1-11, 89-T-26-17 and 90-F-45-14, exhibited no haemolytic activity and no insecticidal activity against dipteran and lepidopteran insects, but were highly cytocidal against leukaemia T cells and other human cancer cells, showing different toxicity spectra and varied activity levels. Furthermore, the proteins from 84-HS-1-11 and 89-T-26-17 were able to discriminate between leukaemia and normal T cells, specifically killing the former cells. These findings may lead to the use of B. thuringiensis inclusion proteins for medical purposes.

Animals↗
PubMed1999

Unique synthetic peptides stimulating streptolysin S production in streptococci.

A peptide has been isolated from pronase digest of bovine serum albumin as the stimulatory factor of streptolysin S (SLS) production by Streptococcus pyogenes, and its primary structure has been deduced [Akao et al. (1992) Infect. Immun. 60, 4777-4780]. To determine the essential structure for the stimulation, a peptide (P-1) having the deduced structure, in which three peptide fragments are linked by two disulfide bonds, and shorter analogs (P-2 to P-4) of peptide P-1 were chemically synthesized. Another peptide (P-5), in which Ala is inserted between the two Cys residues in the middle peptide chain of P-1, was also synthesized. These synthetic peptides were identified by mass spectrometry and analysis of amino acid compositions. The synthetic P-1 stimulated SLS production in a dose-dependent manner. Other peptide analogs also showed remarkable stimulation of SLS production. Treatment of P-1 with performic acid resulted in loss of its stimulatory activity, indicating that disulfide bridges of the peptides are necessary for their activity on SLS production. These results suggest that the unique primary structure of three peptide chains linked by two disulfide bridges is requisite for the stimulatory effect on SLS production.

Amino Acid Sequence↗
PubMed1999

Influence of various bile acids on the metabolism of glycyrrhizin and glycyrrhetic acid by Ruminococcus sp. PO1-3 of human intestinal bacteria.

Ruminococcus sp. PO1-3, an intestinal bacterium isolated from human feces, metabolized glycyrrhizin (GL) to glycyrrhetic acid (GA) and GA to 3-oxo-glycyrrhetic acid (3-oxo-GA) and possessed GL beta-D-glucuronidase and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) involved in the metabolism of GL. This bacterial growth was enhanced by GL at a concentration of 0.4 mm and was suppressed by GA at concentration of 1.0 mM. Chenodeoxycholic acid, deoxycholic acid and lithocholic acid among the bile acids added to this bacterium suppressed the growth and GL beta-D-glucuronidase activity and 3beta-HSD activity incident to it at a concentration of 1.0 mM, while cholic acid, hyodeoxycholic acid and glycine and taurin conjugates of cholic acid, chenodeoxycholic acid, deoxycholic acid and lithocholic acid had almost no effect on this bacterium at a concentration of 0.2 to 1.0 mm. However, these enzyme activities of this sonicated bacteria were inhibited by all of these bile acids. Although each bile acid and GL added to bacteria at the same time suppressed the growth and the amount of metabolite GA by all bile acids used except cholic acid, taurocholic acid and taurodeoxycholic acid with GL, a combination of each bile acid and GA eased the growth inhibition caused by GA at a concentration of 0.2 mM and enhanced the amount of metabolite 3-oxo-GA by the glycine conjugate of bile acids with GA. GL or GA added after 6 h culture with each of these bile acids and bacteria was metabolized to a relatively large amount of GA by chenodeoxycholic acid and lithocholic acid and their glycine and taurine conjugates, glycocholic acid and taurodeoxycholic acid, or had almost no effect on the amount of metabolite 3-oxo-GA, respectively. These results showed that although GL added after the exposure to bile acid and GA and bile acid added at the same time as bacteria had different bile acid action, these conditions enhanced the amount of metabolite GA from GL and metabolite 3-oxo-GA from GA.

Bacteria, Anaerobic↗
PubMed1999

Purification and characterization of glycyrrhetic acid mono-glucuronide beta-D-glucuronidase in Eubacterium sp. GLH.

Glycyrrhetic acid mono-glucuronide (GAMG), 1-(18beta-glycyrrhet-3-yl)-beta-D-glucopyranuroic acid, was hydrolyzed to glycyrrhetic acid (GA) by GAMG beta-D-glucuronidase in Eubacterium sp. GLH from human intestinal bacteria. The enzyme had an optimum pH of 5.0 and was purified from a crude extract by Butyl Toyopearl 650 S, Toyopearl HW-55 S, Hydroxyapatite and DEAE-Toyopearl 650 M column chromatography. The purified enzyme showed a specific activity of 495 nmol/min/mg protein and a single band on Coomassie brilliant blue staining and a molecular weight of about 43 kDa on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The apparent molecular weight was 49.5 kDa, as estimated by Toyopearl HW-55 S column chromatography. Also, the enzyme seemed to have a sulfhydryl group(s) in its active site with a Km value of 77 x 10(-3) M.

Chromatography, Gel↗
PubMed1998

Clonal and chronological genetic analysis of multifocal cancers of the bladder and upper urinary tract.

Recent molecular genetic studies have suggested that multifocal urothelial cancers are derived from an identical progenitor cell. However, the clonal origin of multifocal urothelial cancers of a low-grade superficial type has not been fully defined. Using microsatellite markers, we examined genetic alterations at 20 loci on eight chromosomal arms (2q, 4p, 4q, 8p, 9p, 9q, 11p, and 17p) in 87 metachronous and/or synchronous multifocal urothelial cancers, which included 84 low-grade superficial papillary tumors from 29 patients. Judging from the patterns of loss of heterozygosity, microsatellite shifts, and the subchromosomal partial deletion, multifocal tumors in at least 20 (80%) of the 25 evaluable patients were considered to be derived from a single progenitor cell, although the possibility remained that multifocal tumors in a small subset of patients might develop from distinct progenitor cells due to field cancerization. In 13 of the 20 patients, a chronological genetic analysis was available: genetic heterogeneity was detected in 3 (23%) patients, and an apparent accumulated pattern of genetic alterations was detected in only 1 (8%) patient. In the 20 patients with multifocal tumors of an identical clonal origin, discordant microsatellite alterations were observed, with significantly lower frequencies on chromosome 9 compared to those on the other chromosomes tested. The results indicate that most multifocal low-grade superficial urothelial cancers are genetically stable despite their incidence of frequent recurrence, and genetic divergence occurs in a subset of patients. This heterotopic spread and genetic divergence may occur long before the clinical manifestation of multiplicity from a single transformed cell. These data support the previous view that heterotopic spread of transformed progenitor cells and genetic divergence occur after chromosome 9 alterations in most of low-grade superficial urothelial cancers.

Chromosome Aberrations↗
PubMed1998

Unique appendages associated with spores of Bacillus cereus isolates.

Electron microscopic observations revealed the presence of a new type of large appendage on the spores of two Bacillus cereus strains isolated from phylloplanes. The appendages were thin and sword-like in shape, having the sizes of 1.5 to 2.8 microns in length and 0.03 to 0.6 micron in width. There were no core or sheath structures in these appendages. The number of appendages on a spore ranged from three to more than twenty, radiating from the swelling on one end of the exosporium. These appendages gave a unique octopus- or jellyfish-like feature to the spores.

Bacillus cereus↗