Detection of circulating lymphoma cells in subcutaneous panniculitis-like T-cell lymphoma.
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Aged↗
Biomedical subjects
R Osawa
Publications and source records attributed to R Osawa.
[Effect of probucol on the concentration of cyclosporin A in patients with nephrotic syndrome].
The first choice of therapy for nephrotic syndrome is steroid, and cyclosporin A(CyA) or other immunosuppressants are selected for steroid resistant or recurrent cases. Nephrotic syndrome accompanies hyperlipidemia for which HMG-CoA reductase inhibitors are mainly used. On the other hand, probucol is used in cases showing inadequate effects or some adverse reactions under treatment with HMG-CoA reductase inhibitors. Recently, we experienced several cases whose blood levels of CyA were decreased to about half that before the combined use of probucol, and concomitant administrations were discontinued. Based on these cases, we considered that the use of probucol should be prescribed in patients with nephrotic syndrome accompanying hyperlipidemia giving preference to CyA treatment. In cases of unavoidable usage of probucol, CyA dose adjustments are required on the basis of frequent CyA blood level monitoring.
Adult↗
Isolation of tannin-degrading lactobacilli from humans and fermented foods.
Lactobacilli with tannase activity were isolated from human feces and fermented foods. A PCR-based taxonomic assay revealed that the isolates belong to Lactobacillus plantarum, L. paraplantarum, and L. pentosus. Additional studies on a range of Lactobacillus species from established culture collections confirmed that this enzymatic activity is a phenotypic property common to these three species.
Animals↗
Growth of starved Escherichia coli O157 cells in selective and non-selective media.
Escherichia coli O157 strains starved in sterile deionized water (SDW) and filter-sterilized natural river water (SRW) were investigated with specific reference to their culturability in selective and non-selective media. Growth of the strains starved in both SDW and SRW were markedly suppressed with time in selective liquid media such as modified trypticase soy broth supplemented with novobiocin (mTSB+n) and modified E. coli broth supplemented with novobiocin (mEC+n). This suppression was more pronounced when incubated at 42 C than at 37 C, especially with mEC+n. By contrast, such growth suppression was seldom observed when cultured at 37 C in non-selective liquid media such as trypticase soy broth (TSB) and buffered peptone water. In mEC+n at 42 C, the non-starved cells from overnight cultures with an initial density of less than 10(3) colony-forming units (CFU)/ml grew to the density of over 10(7) CFU/ml after 24 hr incubation, whereas those starved for 6 weeks in SRW were only to maintain their initial density or died off after 24 hr incubation under the same culturing conditions. These results indicated that the isolation of starved cells of E. coli O157 from water samples would be most difficult with selective enrichment or direct plating on the selective plate media. It is thus highly recommended that a "resuscitation" of the cells with non-selective enrichment should be performed as a routine practice for maximum recovery of E. coli O157 from water systems.
Animals↗
[Isolation of Shiga toxin-producing Escherichia coli O157:H7 from processed salmon roe associated with the outbreaks in Japan, 1998, and a molecular typing of the isolates by pulsed-field gel electrophoresis].
Shiga toxin-producing Escherichia coil (STEC) O157 were isolated from processed salmon roe which had been a suspected food item in sporadic infections which occurred in Japan in 1998. A total of 45 samples of the processed salmon roe were pre-enriched in trypticase soy broth (TSB) at 36 degrees C for 6 h and novobiocin-supplemented modified EC broth (mEC-NB) at 42 degrees C for 18 h. After the pre-enrichments, the cultures were examined for possible occurrence of STEC O157, using an immunomagnetic separation (IMS) method. From the examination, a total of 84 strains of STEC O157:H7 that were positive for both stx 1 and stx 2 genes were isolated. By applying the most-probable-number technique, it was estimated that the number of STEC O157 was in the range of 0.73-1.5 per 10 g of the processed salmon roe. Subsequent analysis of the isolates by a pulsed-field gel electrophoresis (PFGE) revealed a pattern commonly seen in 82 isolates and another pattern in two isolates. Clinical isolates from 7 patients also showed an identical pattern to those of the 82 isolates and one isolate from a patient showed the other pattern identical to those of the two isolates. The isolates were found to belong to the phage type 14.
DNA, Bacterial↗
Characteristics of Streptococcus pyogenes serotype M1 and M3 isolates from patients in Japan from 1981 to 1997.
Streptococcus pyogenes isolates obtained in 1981 to 1997 from patients and healthy subjects were characterized by pulsed-field gel electrophoresis (PFGE) patterns, biotyping, and the presence of spe genes encoding streptococcal pyrogenic exotoxins. Changes in the profiles were shown in the serotype M1/T1 isolates from pharyngitis over this period, but not in serotype M3/T3 isolates. The characteristics of isolates from patients with toxic shock-like syndrome (TSLS) were comparable to those of the other isolates, including those from healthy subjects. This finding suggests that further phenotypic and molecular characterization, such as investigating the genomic difference represented by the pathogenicity island, of isolates with apparently the same profiles would be necessary to determine the etiology of diseases caused by S. pyogenes, including TSLS.
Adolescent↗
[A comparison of sensitivity between direct plate culture, immunomagnetic separation and polymerase chain reaction for the isolation of enterohemorrhagic Escherichia coli O157].
Sensitivities of direct plate culture (DPC) method, immunomagnetic separation (IMS) method, and polymerase chain reaction (PCR) assay for successful detection Escherichia coli O157 in the food samples were compared. Three lots of minced beef and three lots of radish sprout, both of which were commercially retailed, were enriched with non-selective broth media at 36 degrees C for 6 h. After enrichment, the cultures of the minced beef and those of the radish sprout were found to have background microflora at ca.10(5)-10(7) CFU/ml and ca.10(8) CFU/ml, respectively. The cultures were then experimentally inoculated with E. coli O157 strains at various final concentrations ranging from ca.10 to 10(7) CFU/ml. The samples thus prepared were subjected to the above three methods to evaluate their detection limits. For the samples of minced beef, the detection limits of the DPC method was 10(2) CFU/ml whilst that of the IMS method was ca.10 CFU/ml. For the samples of radish sprout, the detection limits of the DPC method, the IMS method, and the PCR assay were ca.10(4) CFU/ml, ca.10(2) CFU/ml, and ca.10(6) CFU/ml, respectively. There results strongly suggest that the IMS method is most sensitive method for the detection of O157 from food samples among the methods currently available.
Bacteriological Techniques↗
Urea hydrolysis and suppressed production of thermostable direct hemolysin (TDH) by Vibrio parahaemolyticus associated with presence of TDH-related hemolysin genes.
A total of 18 strains of V. parahaemolyticus isolated from patients of past food poisoning cases occurring in Kanagawa Prefecture, Japan, were assayed for presence of the thermostable direct hemolysin (TDH) gene and the TDH-related hemolysin (TRH) genes (trh 1 and trh 2) with specific reference to their ability to hydrolyze urea and TDH production. A polymerase chain reaction assay revealed that all urea-hydrolyzing strains (9 strains) carried either trh 1 gene or trh 2 gene. The strains carrying the trh genes as well as the tdh gene produced TDH less by a factor of 4 to 16 than those carrying only the tdh gene, suggesting the expression of the tdh gene was suppressed by the presence of trh gene through a mechanism yet to be defined.
Bacterial Proteins↗
Digestion of staphylococcal enterotoxin by Bacillus natto.
Cooked rice contaminated with staphylococcal enterotoxin A (SEA) was mixed with 'natto', a Bacillus natto fermented soybean food, and the mixture was incubated at 37 degrees C for 1 h. Reversed passive latex agglutination (RPLA) tests performed on the mixture revealed that the RPLA titer against SEA was significantly reduced after incubation. Subsequent analytical tests showed that the SEA protein molecule was fragmented to small peptides by an extracellular protease, subtilisin, produced by B. natto. The proteolytic activity of B. natto was also found to be effective against other types of staphylococcal enterotoxins.
Agglutination Tests↗
The tannin-degrading species Streptococcus gallolyticus and Streptococcus caprinus are subjective synonyms.
The tannin-degrading species Streptococcus gallolyticus and Streptococcus caprinus have been shown to be subjective synonyms on the basis of their levels of 16S rRNA sequence similarity (98.3%) and DNA-DNA homology (> 70%) and the phenotypes of their type strains. S. gallolyticus has nomenclatural priority according to Rule 24b(2) of the International Code of Nomenclature of Bacteria.
Animals↗
[Isolation of enterohemorrhagic Escherichia coli (O157:H7) by an immunomagnetic separation method].
Three sporadic cases of enterohemorrhagic Escherichia coli (EHEC) O157 infection which occurred in Kanagawa in 1996 were investigated. In an attempt to determine sources of the infection, a novel method of immunomagnetic separation (IMS) was employed to isolate the bacterium from feces, foods, and other associated items. In the first case, strains of EHEC O157:H7 producing Vero toxin (VT) 2 were isolated from both feces of the patient and suspected food (cattle liver) kept at a restaurant, and the strains were found to be genotypically identical through an analysis of pulsed-field gel electrophoresis (PFGE). Subsequent investigation in the meat processing store, from which the above cattle liver had been retailed to the restaurants revealed that the store was contaminated with EHEC O157:H7 producing both VT1 and VT2. In the second case, a strain isolated from the patient was EHEC O157:H7 producing both VT1 and VT2 while strains isolated from the patient's family (without apparent symptom) and the suspected facility were O137:NM producing VT2. PFGE analysis indicated that the latter two strains were genotypically identical, suggesting that the facility thus contaminated with EHEC O157 caused the infection in question. In the third case, EHEC O157:NM producing VT2 was isolated from 4 out of 7 family members including the patient, and these strains were found to be genotypically identical by subsequent PFGE analysis. Source of the infection was, however, not determined due to lack of suspected food items. In this context, four slaughterhouses in Kanagawa Prefecture were investigated for presence of EHEC O157. As a result, strains of EHEC O157:H7 producing VT1 and VT2 were isolated from the contents of cattle's distal colon and surface of the skinned carcasses. Additional attempt was also made to determine a possibility of river water being contaminated with EHEC O157. The bacterium was, however, not isolated from water samples collected from 4 major rivers in the prefecture (at 10 collecting sites). Experiments were undertaken in order to evaluate the use of IMS in isolation of EHEC O157 from food items, with different pre-enrichment media and conditions. The results indicated that pre-enrichment by trypticase soy broth at 36 degrees C for 6 h followed by inoculating onto sorbitol MacConkey agar plate containing cefixime and tellurite was most appropriate to isolate EHEC O157 strains.
Adolescent↗
Rapid screening method for identification of cholera toxin-producing Vibrio cholerae O1 and O139.
A novel method of identifying cholera enterotoxin (CT)-producing Vibrio cholerae serogroups O1 and O139 was developed. The method uses degradation of NAD as a specific biochemical marker for the CT-producing strains. The substrate NAD at a concentration of 100 mumol/liter was markedly degraded when it was incubated at 37 degrees C for 2 h with the CT-producing stains at a final cell density equivalent to that of a twofold dilution of a McFarland no. 1 standard. NAD degradation was monitored by an enzyme-amplified color development assay. Subsequent tests conducted with a total of 119 strains of V. cholerae, including both clinical and environmental isolates, confirmed a significant correlation between NAD degradation and CT production for all V. cholerae strains belonging to serogroups O1 and O139. Since 2 of 11 non-O1, non-O139 V. cholerae strains not carrying the CT gene degraded NAD, serotyping of the strains prior to the test is recommended.
Bacterial Typing Techniques↗
Highly expressed human sialyl Lewis antigen on cell surface of streptococcus gallolyticus.
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Animals↗
Occurrence of urease-positive Vibrio parahaemolyticus in Kanagawa, Japan, with specific reference to presence of thermostable direct hemolysin (TDH) and the TDH-related-hemolysin genes.
A total of 132 strains of V. parahaemolyticus isolated from patients and from the suspected causal food items of past food poisoning cases occurring in Kanagawa Prefecture, Japan, were examined for the ability to hydrolyze urea, with specific reference to the presence of the thermostable direct hemolysin gene (tdh) and the gene for thermostable direct hemolysin-related hemolysin (trh). Ten strains belonging to five different O-antigen serotypes were positive for urea hydrolysis (UH+), and four of these strains did not carry tdh. A total of 106 strains carried tdh, but less than 6% of them were UH+, whereas all trh-carrying strains were UH+. The evidence suggests that urea hydrolysis is not a reliable marker for identifying tdh-carrying V. parahaemolyticus strains in Japan (the Pacific Northeast) but may be a marker for trh-carrying strains.
Bacterial Proteins↗
Production of thermostable direct hemolysin by Vibrio parahaemolyticus enhanced by conjugated bile acids.
The effects of conjugated bile acids, glycocholic acid, and taurocholic acid (TC) on production of thermostable direct hemolysin (TDH) by Vibrio parahaemolyticus were determined by a reversed passive latex agglutination assay against TDH. The amount of TDH excreted in growth medium containing either glycocholic acid or taurocholic acid (5 mM/liter) was, on a per-cell basis, 4- to 16-fold greater than that excreted in medium without the bile acids. The amounts of TDH released from lysed cells grown with the bile acids (5 mM/liter) were 4- to 32-fold greater than those from lysed cells grown without, suggesting that the bile acids enhanced synthesis of TDH within bacterial cells. These data imply that the conjugated bile acids play a key role in the pathogenicity of V. parahaemolyticus.
Glycocholic Acid↗
Effects of different acidulants on growth of 'Streptococcus milleri group' strains isolated from various sites of the human body.
Growth of human clinical isolates of Streptococcus constellatus, Strep. intermedius and Strep. anginosus in HCl-, acetate and lactate acidified media was investigated. Under aerobic conditions, Strep. constellatus and Strep. anginosus were significantly more tolerant to all the acidulants than was Strep. intermedius. Under anaerobic conditions, Strep. anginosus and Strep. intermedius were significantly more tolerant to acetic acid (pH < 4.5) than Strep. constellatus.
Acetates↗
Biochemical and antigenical characterization of tannin-protein complex degrading enterobacteria isolated from koalas, Phascolarctos cinereus.
Biochemical and antigenical characteristics of tannin-protein complex degrading enterobacteria (T-PCDE) isolated from Koalas, Phascolarctos cinereus, were investigated. T-PCDE had a specific profile of characteristics, and T-PCDE was distinguished from those of 12 type strains of Enterobacteriaceae used.
Animals↗
An investigation of gram-negative tannin-protein complex degrading bacteria in fecal flora of various mammals.
Gram-negative tannin-protein complex degrading bacteria (T-PCDB) were first isolated from animals except for the koala. The occurrence of T-PCDB in feces of 15 species of mammals with different feeding habits was investigated. T-PCDB occurred in 7 of 54 horses but they could not be isolated from other mammals tested. These T-PCDB comprised less than 0.1% of the facultative anaerobic microflora in horse feces and it was much less than that previously reported in koala feces ( > 60%). A total of 7 T-PCDB fecal isolates showed a range of phenotypic diversities. They were all Gram-negative rods of various sizes and shapes including coccoidal rod. Although all produced tannase, no strain showed to have gallate decarboxylase. A total of 23 representative strains belonging to the family Pasteurellaceae were also tested for tannase production. Two strains, Haemophilus actinomycetemcomitans NIAH-10202T and Haemophilus segnis NIAH-10183T which were isolated from human oral cavity were detected positive for tannase.
Animals↗