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Biomedical subjects

K Kuroiwa

Publications and source records attributed to K Kuroiwa.

At least 19 recordsLinked to original sources

Elevation of ST2 protein levels in cerebrospinal fluid following subarachnoid hemorrhage.

OBJECTIVE: To study the mode of appearance of ST2 in cerebrospinal fluid (CSF) after subarachnoid hemorrhage (SAH). MATERIALS AND METHODS: Immunoprecipitation and subsequent immunoblotting were performed to reveal the existence of ST2 in CSF after SAH. CSF samples from 21 patients were analyzed for ST2 using an enzyme-linked immunosorbent assay system. The ST2 levels were compared between serum and CSF after SAH. The ST2 levels in CSF were measured in six patients operated with other than SAH. RESULTS: ST2 was secreted into CSF after SAH. The concentration of ST2 was the highest in the samples of the first post-operative day and declined thereafter. The patients operated with other than SAH did not show the elevation of ST2 in CSF. CONCLUSIONS: This study revealed the presence of ST2 in CSF for the first time and suggested a possibility that ST2 is related to the inflammatory reaction in the central nervous system after SAH.

Adult↗

Augmenting effect of acetic acid for acidification on bactericidal activity of hypochlorite solution.

AIMS: Bactericidal activity of chlorine solution is enhanced by weak acidification. We compared the effects of various acids on the bactericidal activity of hypochlorite solution to establish a method for safe and effective use of an acidic hypochlorite solution. METHODS AND RESULTS: The bactericidal activities of acidic hypochlorite solutions that had been adjusted to pH 5.0 with hydrochloric acid, acetic acid, citric acid, lactic acid, formic acid, phosphoric acid or sulphuric acid against Bacillus subtilis spores were compared. The acidic solutions prepared with hydrochloric acid and acetic acid showed the highest bactericidal activity, and all of the spores (5 x 106 cfu ml(-1)) were killed within 10 min. On the other hand, the solutions prepared with citric acid and lactic acid showed no bactericidal activity against any bacterial strains tested in this study despite the low pH. The amount of chlorine gas produced by the preparation using acetic acid was sixfold less than that produced from the preparation using hydrochloric acid. CONCLUSIONS: Acetic acid is the most suitable and safe acid for the preparation of an acidic hypochlorite solution. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study provide useful information for establishing a method for safe and effective use of an acidic hypochlorite solution.

Acetic Acid↗

Renal cell carcinoma with rhabdoid features: an aggressive neoplasm.

AIMS: Only a few reports on renal cell carcinoma with rhabdoid features have been published. This study was performed to investigate the clinicopathological characteristics of renal cell carcinomas with rhabdoid features. METHODS AND RESULTS: Among 253 cases of renal cell carcinoma in adults, eight cases with rhabdoid features were detected. Rhabdoid areas ranged from 10% to 90% of each of the cases. Seven of the eight cases were TNM stage III or IV, and four of the eight cases died within 8 months of surgery. Immunohistochemically, the rhabdoid areas were positive for CAM 5.2 (4/8), AE1/AE3 (6/8), epithelial membrane antigen (6/8) and vimentin (8/8), and negative for myogenetic markers (0/8). The mean MIB-1 labelling index in the rhabdoid areas was higher than that in the definite carcinomatous areas. Ultrastructurally, perinuclear whorls of intermediate filaments were demonstrated in three of the eight cases using paraffin-embedded blocks. CONCLUSIONS: The rhabdoid areas in renal cell carcinoma have histological, immunohistochemical and ultrastructural similarities to malignant rhabdoid tumours. Renal cell carcinoma with rhabdoid features is a highly aggressive neoplasm and its malignant behaviour may be due to the high cell-proliferative activity of the rhabdoid areas. Rhabdoid features in renal cell carcinoma may represent the endpoint of clonal evolution of renal cell carcinoma (especially in clear cell type cases).

Aged↗

Elevated soluble ST2 protein levels in sera of patients with asthma with an acute exacerbation.

Previous studies have reported that ST2 is preferentially expressed on Th2 cells and plays a critical part in controlling airway inflammation in murine models of asthma. However, the clinical role of ST2 in patients with bronchial asthma remains unclear. In our study, we examined 56 patients with atopic asthma in a nonattack phase and 200 nonatopic normal volunteers for healthy control, and analyzed the relationship of their serum ST2 levels to asthma severity, pulmonary function, and laboratory data. Of the 56 patients with atopic asthma, 30 exhibited asthmatic exacerbation, and their serum ST2 levels were also analyzed. The serum ST2 levels were low, but a statistical difference was found between patients with nonattack asthma and the healthy control group (p < 0.05). We also found a differential rise of serum ST2 level that correlates well with the severity of asthma exacerbation. Furthermore, the serum ST2 levels during asthma exacerbation statistically correlated with the percentage of predicted peak expiratory flow (r = -0.634, p = 0.004) and Pa(CO(2)) (r = 0.516, p = 0.003). These results suggest that soluble human ST2 protein in sera may be related to Th2-mediated allergic inflammation inducing acute exacerbation in patients with atopic asthma.

Acute Disease↗

Identification of human ST2 protein in the sera of patients with autoimmune diseases.

Soluble human ST2 protein (IL1RL1-a) in the sera of patients with various autoimmune diseases was identified by a newly developed procedure using specific monoclonal antibodies. After immunoprecipitation and subsequent immunoblotting, a glycosylated protein of about 60 kDa was detected in the sera of SLE patients, but not in the sera of healthy controls. The experiments using gel filtration and SDS-PAGE under a nonreducing condition indicated the existence of the ST2 multimer in serum. The mobility of the natural protein was slower than that of the recombinant human ST2 protein produced by COS7 cells in SDS-PAGE, suggesting a difference of glycosylation between humans and monkeys. The identification of the natural human ST2 protein should be important both to fundamental researches and the further clarification of the clinical implications of the ST2 protein.

Animals↗

Cell proliferative activity and expression of cell-cell adhesion factors (E-cadherin, alpha-, beta-, and gamma-catenin, and p120) in sarcomatoid renal cell carcinoma.

BACKGROUND AND OBJECTIVES: Sarcomatoid RCC (renal cell carcinoma) is an uncommon, but not rare, neoplasm which has been shown to have a much worse prognosis than common RCC. The current study was designed to investigate the association of proliferative activity and cell-cell adhesion molecules with sarcomatoid RCC. METHODS: Proliferative activity (Ki-67 labeling index) and expression of cell-cell adhesion associated molecules (E-cadherin, alpha-, beta-, and gamma-catenin, and p120) were examined using immunohistochemical techniques in 11 cases of sarcomatoid RCC. RESULTS: In six patients with sarcomatous component more than 50%, five were died within 24 month after diagnosis. The expression of these molecules within the carcinomatous and sarcomatous components of sarcomatoid RCC was compared. The mean Ki-67 labeling index in the sarcomatous components (12.6%) is statistically higher than in the carcinomatous components (3.7%) (P < 0.05). The expressions of E-cadherin, and alpha-, beta-, and gamma-catenin were statistically decreased in the sarcomatous components compared to the carcinomatous components. However, no differences were observed regarding p120 immunostaining. CONCLUSIONS: The findings of the current study suggest that the range of the sarcomatous component may be a prognostic factor in RCC, and the malignant behavior of sarcomatoid RCC is due to high cell proliferative activity and decreased expressions of E-cadherin and alpha-, beta-, and gamma-catenin in the sarcomatous component.

Aged↗

Presence of a genistein-responsive inhibitory mechanism on interleukin-1alpha-induced NF-kappaB activation.

Interleukin 1 (IL-1) is known to activate the signal transduction machinery, including the transcription factor, nuclear factor kappa B (NF-kappaB). The activation mechanism of NF-kappaB has been studied intensively, while the negative regulatory mechanisms of NF-kappaB remain to be clarified. In the present study, we found that genistein, a tyrosine kinase inhibitor, augmented IL-1alpha-dependent NF-kappaB activation, suggesting the presence of a tyrosine kinase mediating a suppression signal on NF-kappaB. As determined by luciferase reporter gene assay using kappaB-responsive element, genistein enhanced IL-1alpha-induced NF-kappaB activation. Although genistein failed to increase luciferase activity at 1 and 3 h after IL-1alpha stimulation, it induced prolonged activation beginning at 6 h after the initial stimulation. We next examined whether genistein augmented the DNA-binding activity of NF-kappaB, using electrophoretic mobility shift assay. In the case of the control experiment, the binding of NF- kappaB to the kappaB-responsive element peaked at 30 min after IL-1alpha stimulation, and decreased thereafter. In contrast, treatment with genistein maintained the maximum binding activity for at least 2 h after stimulation. Moreover, genistein enhanced the IL-1alpha-dependent degradation of IkappaBalpha. Taken together, our results indicate that genistein augments IkappaB degradation, resulting in continuous NF-kappaB activation. This suggests the possibility that tyrosine kinase negatively regulates NF-kappaB.

Base Sequence↗

Cloning and characterization of a chitosanase gene from the koji mold Aspergillus oryzae strain IAM 2660.

A genomic copy of the gene coding for chitosanase (csnA) was isolated from Aspergillus oryzae IAM 2660. A. oryzae csnA contains an open reading frame that encodes a polypeptide of 245 amino acids with a calculated molecular mass of 26,500 Da. The deduced amino acid sequence of A. oryzae csnA indicates extensive similarities to those of other fungal chitosanases.

Amino Acid Sequence↗

The cloning and nucleotide sequence of human ST2L cDNA.

The ST2 gene is a member of the IL-1 receptor family and is hypothesized to be involved in helper T cell function, but its functional ligand and physiological role remain unknown. We have cloned the human ST2L cDNA that encodes a distinct type of membrane-bound ST2 protein. The predicted 556-amino-acid sequence showed 67% identity to the mouse ST2L protein. The human ST2 gene (IL1RL1) contains 13 exons and spans 40 kb in length. Its exon-intron organization was elucidated from a registered human genomic sequence derived from chromosome 2q, which contains three other genes belonging to the IL-1 receptor family in an approximately 202-kb genomic region. The tissue distribution of ST2 expression was examined by RT-PCR, and the soluble form (ST2, IL1RL1-a) and ST2L (IL1RL1-b) appear to be expressed differentially. We also established stable transfectants of a human glioblastoma cell line, T98G, that express human ST2L constitutively, and we confirmed cell-surface expression of human ST2L protein on the transfectants.

Amino Acid Sequence↗

Distribution of Pb, Cd and Ba in soils and plants of two contaminated sites.

Evaluation of metal accumulation in soils and plants is of environmental importance due to their health effects on humans and other biota. Soil material and plant tissue were collected along transects in two heavily contaminated facilities, a Superfund site and a lead-acid battery dump, and analyzed for metal content. Soil lead (Pb), cadmium (Cd) and barium (Ba) concentrations for the Superfund site averaged 55,480, 8.5 and 132.3 mg/kg, respectively. Soil Pb occurred primarily in the carbonate, sulfide/residual and organic chemical fractions (41.6, 28.6 and 16.7%, respectively). Soil Pb, Cd and Ba concentrations for the dump site averaged 29,400, 3.9 and 1130 mg/kg, respectively. Soil Pb occurred mostly in the organic and carbonate fractions as 48.5 and 42.5%, respectively. Pb uptake in the two sites ranged from non-detectable (Agrostemma githago, Plantago rugelii, Alliaria officinalis shoots), to 1800 mg/kg (Agrostemma githago root). Cd uptake was maximal in Taraxacum officinale at 15.4 mg/kg (Superfund site). In the majority > or =65%) of the plants studied, root Pb and Cd content was higher than that for the shoots. Tissue Pb correlated slightly with exchangeable and soluble soil Pb; however, tissue Cd was poorly correlated with soil Cd species. None of the sampled plants accumulated measurable amounts of Ba. Those plants that removed most Pb and Cd were predominantly herbaceous species, some of which produce sufficient biomass to be practical for phytoremediation technologies. Growth chamber studies demonstrated the ability of T. officinale and Ambrosia artemisiifolia to successfully remove soil Pb and Cd during repeated croppings. Tissue Pb was correlated with exchangeable soil Pb at r(2)=0.68 in Ambrosia artemisiifolia.

Journal Article↗

Construction of ELISA system to quantify human ST2 protein in sera of patients.

The human ST2 gene can be specifically induced by growth stimulation in fibroblastic cells, and can also be induced by antigen stimulation in Th2 cells. The gene encodes a soluble secreted protein, ST2, and a transmembrane protein, ST2L, which are closely related to the interleukin-1 receptor. To gain insight into the biological roles of the ST2 gene, three monoclonal antibodies (MAbs) against human ST2 gene products were obtained. To obtain these antibodies, immunization was carried out using two different immunogens: purified soluble human ST2 protein (hST2), and COS7 cells, which express the extracellular portion of human ST2L. 2A5 and FB9 MAbs were derived from the immunization with soluble hST2, and HB12 was derived from the COS7 cell immunization. All three antibodies were shown to detect native forms of the human ST2 gene products by immunoprecipitation, flow cytometry, and enzyme-linked immunosorbent assay (ELISA). In the competitive ELISA using biotinylated and nonlabelled MAbs, neither FB9 nor HB12 affected the binding of 2A5 to ST2 gene products. Based on this result, we constructed a sandwich ELISA system using 2A5 and FB9 to measure the concentration of soluble hST2 in sera. The ELISA, combined with the flow cytometry using these antibodies, will be a useful tool for elucidating the functions of human ST2 gene products in individuals.

Animals↗

Purification and characterization of chitosanase and Exo-beta-D-glucosaminidase from a Koji mold, Aspergillus oryzae IAM2660.

Chitosan-degrading activity was detected in the culture fluid of Aspergillus oryzae, A. sojae, and A. flavus among various fungal strains belonging to the genus Aspergillus. One of the strong producers, A. oryzae IAM2660 had a higher level of chitosanolytic activity when N-acetylglucosamine (GlcNAc) was used as a carbon source. Two chitosanolytic enzymes, 40 kDa and 135 kDa in molecular masses, were purified from the culture fluid of A. oryzae IAM2660. Viscosimetric assay and an analysis of reaction products by thin-layer chromatography clearly indicated the endo- and exo-type cleavage manner for the 40-kDa and 135-kDa enzymes, respectively. The 40-kDa enzyme, designated chitosanase, catalyzed a hydrolysis of glucosamine (GlcN) oligomers larger than pentamer, glycol chitosan, and chitosan with a low degree of acetylation (0-30%). The 135-kDa exo-beta-D-glucosaminidase,enzyme,named released a single GlcN residue from the GlcN oligomers and chitosan, but did not release GlcNAc residues from either GlcNAc oligomer or colloidal chitin.

Aspergillus oryzae↗

[Risk factors of anastomotic leak following operations for gastric cancer in the elderly].

The risk factors of anastomotic leak in the elderly following operations for gastric cancer were evaluated by multiple logistic model analysis. Data were taken from 705 operations over a 14-year period. The mean age of patients was 75.8 +/- 7.6 years. The significant risk factors for anastomotic leak were amounts of intra-operative bleeding and male gender. No other factors were significant, including age, preoperative associated diseases, preoperative nutritional states and postoperative complications, some of which were, however, significant factors by univariate analyses. We conclude that we should make every endeavor to lower the amount of intra-operative bleeding in order to prevent postoperative anastomotic leaks in the elderly, especially in male patients.

Adult↗

MDR1 gene overexpression and altered degree of methylation at the promoter region in bladder cancer during chemotherapeutic treatment.

Overexpression of the multidrug resistance 1 (MDR1) gene is closely associated with the clinical outcome of hematopoietic malignancies, but the alteration of its expression during chemotherapeutic treatment and the precise mechanism underlying MDR1 gene overexpression in solid tumors remains unclear. We determined the expression and degree of methylation at the promoter of the MDR1 gene in bladder cancer. The mRNA levels of the MDR1 gene were found to be markedly enhanced, 3.5- to 5.7-fold higher in bladder cancers after chemotherapeutic treatment than those in untreated primary tumors. The MDR1 gene was overexpressed in recurrent tumors in 89% of patients who showed rerecurrence, whereas overexpression was observed in 25% of the patients without re-recurrence. A statistically significant inverse correlation existed between MDR1 expression and the methylation of 5'CpG sites at the promoter in patients with bladder cancer after chemotherapeutic treatment, with the degree of methylation at several CpG sites, rather than other specific sites, involved in this regulation. Consistent with the increase in MDR1 expression, the frequency of patients with a hypermethylated promoter decreased to 50 and 17% after intravesical and systemic chemotherapy, respectively. Thus, overexpression of the MDR1 gene might be a prognostic marker for intravesical recurrence, whereas methylation of the promoter region negatively regulates MDR1 expression and the appearance of multidrug resistance mediated by P-glycoprotein in bladder cancers.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Presence and expression of a novel variant form of ST2 gene product in human leukemic cell line UT-7/GM.

A novel variant cDNA from the human ST2 gene other than ST2 or ST2L was identified and tentatively named ST2V. Alternative splicing inserts a new exon which leads to a change in the C-terminal portion of ST2, causing it to gain a hydrophobic tail instead of losing the third immunoglobulin-like domain. ST2V is expressed in human leukemic cell line UT-7 and its sublines UT-7/GM, UT-7/EPO, and UT-7/TPO, in addition to human helper T cell line 5C10. The amount of ST2V mRNA is greatly diminished when UT-7/GM cells are induced to differentiate into either erythroblastic or megakaryoblastic phenotypes. The possible roles of the ST2V in growth and differentiation are intriguing.

Alternative Splicing↗

Presence of ganglia within the prostatic capsule: ganglion involvement in prostatic cancer.

BACKGROUND: The presence of ganglia within the prostatic capsule (capsular ganglia) is a poorly understood phenomenon. If cancer invasion into or around ganglia is identified in a needle biopsy specimen, such a finding may be diagnosed as an extraprostatic extension. In this study, the presence of capsular ganglia was clarified. Furthermore, we discuss the significance of these ganglia in prostatic cancer patients. METHODS: The study group comprised a total of 42 patients, who had all undergone a radical prostatectomy for prostate adenocarcinoma with a relatively small volume. After surgery, the step-sectioned radical prostatectomy specimens were histologically evaluated. RESULTS: In 22 of 42 cases, capsular ganglia were recognized. With respect to the distribution of the capsular ganglia, the ganglia were most frequently observed at the posteriolateral aspect of the prostatic base. Morphologically there were no obvious differences between the capsular and periprostatic ganglia. Moreover, in 3 of 5 cases with cancer involvement into the capsular ganglia, such involvement was not predictive of extraprostatic extension. CONCLUSIONS: The presence of capsular ganglia needs to be clarified in prostatic cancer patients. Our findings therefore suggest that cancer involvement either in or around the ganglia should not be immediately interpreted as indicating an extraprostatic extension, if such a finding is recognized in a needle biopsy specimen.

Adenocarcinoma↗

MR detection of cytoplasmic fat in clear cell renal cell carcinoma utilizing chemical shift gradient-echo imaging.

We attempted to determine whether cytoplasmic fat in clear cell renal cell carcinoma (RCC) can be identified by chemical shift gradient-echo magnetic resonance imaging (CSI). CSI was performed for 22 clear cell RCCs and 30 other renal tumors (including 16 non-clear cell RCCs), all of which were surgically proven. Signal reduction in out-of-phase images of these tumors was retrospectively evaluated and compared. The signal loss ratio (SLR) was defined and calculated. Fat staining of specimens from 16 tumors was performed and correlated with SLR. SLR was significantly higher in clear cell RCCs than in non-clear cell RCCs (P < 0.001). There was a significant correlation between degree of fat staining positivity of the specimens and SLR (P < 0.01). When signal reduction in out-of-phase images suggested the diagnosis of clear cell RCC, correct diagnosis of this entity was made in resected renal tumors with sensitivity, specificity, and accuracy of 82%, 90%, and 87%, respectively. CSI can demonstrate cytoplasmic fat in clear cell RCCs, which helps to differentiate this entity from other RCCs.

Adenocarcinoma, Clear Cell↗