Determination of alpha-amylase using a new blocked substrate (2-chloro-4-nitrophenyl 4(4)-O-beta-D-galactopyranosyl-beta-maltotetraoside)
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Carbohydrate Sequence↗
Biomedical subjects
S Teshima
Publications and source records attributed to S Teshima.
Alteration of the respiratory burst and phagocytosis of macrophages under protein malnutrition.
To understand the role of macrophages in impaired host defense under protein malnutrition (PM), we examined the activities of the respiratory burst and phagocytosis of resident peritoneal macrophages from weaning female mice fed 5% casein or 5% soy protein isolate (SPI) diet for 14 days. Resident macrophages from the low-protein diet groups released larger amounts of superoxide anion (O2-) after stimulation by phorbol 12-myristate 13-acetate (PMA) or opsonized zymosan than those from the 20% casein and 20% SPI diet groups. Activation of macrophages from protein-deficient mice in vitro with lipopolysaccharide (LPS) or macrophage colony-stimulating factor (M-CSF) under LPS-free conditions did not further enhance O2- production. In spite of the increased O2- production with opsonized zymosan, macrophages from protein-deficient mice did not show any acceleration of phagocytosis of Candida albicans in the presence of normal serum. Our results confirm that the phagocytic function of macrophages is susceptible to PM, and suggest that functional alterations of macrophages may be involved in the failure of development of a specific immune response under PM. Furthermore, the enhanced production of oxygen intermediates by macrophages may augment tissue damage under PM.
Animals↗
Prognostic significance of the overexpression of c-erbB-2 protein in adenocarcinoma of the uterine cervix.
BACKGROUND: The overexpression of c-erbB-2 protein is a prognostic marker in patients with breast cancer. Alteration of the c-erbB-2 oncogene in development of adenocarcinoma of the uterine cervix, an unfavorable gynecologic neoplasm, is unknown. METHODS: To clarify the role of the c-erbB-2 oncogene in adenocarcinoma of the uterine cervix, formalin-fixed, paraffin-embedded tissue sections from 44 cases of cervical adenocarcinoma were immunohistochemically examined for expression of c-erbB-2 protein and for c-erbB-2 gene amplification in DNA by slot blot-hybridization analysis. RESULTS: The expression of c-erbB-2 protein was detected in 34 cases (77%). Strong expression on cell membranes was detected in 11 cases (25%). Most cases with strong membrane expression of c-erbB-2 protein also showed amplification of the c-erbB-2 gene by slot blot-hybridization. Expression of the protein on cell membranes was more often seen at clinical Stage II or III (9 of 23 [39%]) than at Stage 0 or 1 (2 of 21 [9%]) (P < 0.05, Fisher's exact test). Expression of the c-erbB-2 protein was also associated with poorer prognosis of patients with cervical adenocarcinoma by comparison of survival curves (P < 0.005) and by the Cox regression model analysis. CONCLUSIONS: Overexpression of c-erbB-2 protein is associated with amplification of c-erbB-2 gene and frequently occurred in cervical adenocarcinomas of the patients with poor prognoses.
Adenocarcinoma↗
Prognostic significance of histopathological subtypes in stage I pure yolk sac tumour of the ovary.
The correlation between histological subtype [endodermal sinus (ES), polyvesicular vitelline (PV), glandular (G) and hepatoid (H) subtypes] and the prognosis of pure yolk sac tumours (YSTs) of the ovary was investigated. From 1964 to 1989, 35 patients with YSTs were treated with primary surgery and adjuvant chemotherapy. The prevalence of histological subtypes was as follows: 14 patients had a single subtype, either ES (12) or G (2); 12 patients had two subtypes, ES+G (4), ES+PV (3), ES+H (4) or G+H (1); six patients had three subtypes, ES+P+H (4) or ES+G+H (2); and three patients had all four subtypes. Multivariate analysis showed that important predictors were FIGO stage, chemotherapeutic regimen and residual tumour size. However, for stage I, multivariate analysis showed that the histological subtype was a superior predictor to the subclassification of FIGO stage I, age or chemotherapeutic regimen (P = 0.03). Kaplan-Meier analysis showed that YSTs composed of an admixture of three or four subtypes was associated with a better prognosis than those composed of one or two subtypes (P < 0.01), other variables being constant.
Adolescent↗
An autopsy case of extraskeletal Ewing's sarcoma followed up for 24 years.
A long-term (24 years) follow-up case of extraskeletal Ewing's sarcoma is reported. The light microscopic examination showed features hardly indistinguishable from Ewing's sarcoma of the bone, that is, the tumor cells were diffusely arranged and uniform in size and shape, and possessed glycogen in the cytoplasm. Homer-Wright rosettes were found only in the autopsy material. An immunohistochemical study using a neural marker (neuron-specific enolase) demonstrated positive staining in most tumor cells. An ultrastructural study revealed intracytoplasmic glycogen particles and incomplete neural characters as follows: the cytoplasmic processes resembled neural processes without neurosecretory granules, microtubules or neurofilaments. These findings suggest that this case finally acquired an incomplete neural character with repeated recurrence. This tumor was diagnosed extraskeletal Ewing's sarcoma, but in future it may be categorized as primitive neuroectodermal tumor (PNET).
Adult↗
[Heterochronic occurrence of bilateral torsion of appendix testis a case report].
An 8-year-old boy who had undergone excision of the left appendix testis for torsion of the left appendix testis about one and a half years previously was brought to our department on February 18, 1991 because of right scrotal pain of 4 days' duration. Palpation revealed induration with tenderness of the superior portion of the right testicle. A scrotal ultrasonographic tomogram revealed a shadow probably representing an enlarged appendix testis. The patient was diagnosed as having torsion of the right appendix testis. There was refractory pain, and there was possibility of reactive epididymitis. So the patient underwent excision of the right appendix testis, two right appendices testis, which had enlarged to 5 mm in diameter were found, histopathological examination revealed slight bleeding and marked edema of the stroma, which may have been caused by torsion of the appendix testis. Scrotal pain subsided postoperatively. Bilateral torsion of the appendix testis is very uncommon, and our case is the 14th case reported. Torsion of the appendix testis occasionally develops. Torsion of the appendix testis occasionally develops bilaterally. When a small, tender mass at the superior pole of testis, torsion of the appendix testis should be considered.
Child↗
Identification of basophils by immunohistochemistry in the airways of post-mortem cases of fatal asthma.
There is increasing evidence for the role of basophils in the pathogenesis of bronchial asthma. To examine the presence of basophils in the airways of patients with fatal asthma by immunohistochemistry, we stained lung tissues from four post-mortem cases who had died from severe asthmatic attacks and four controls with a monoclonal antibody raised against tryptase (AA-1) and anti-IgE. Mast cells and basophils were identified in the bronchioles as AA-1- and anti-IgE-positive cells, and anti-IgE-positive cells, respectively. Airway mast cells were found beneath the basement membrane, near blood vessels in the submucosa, and adjacent to the submucosal glands, and scattered throughout the muscle bundles. There was a significant increase of mast cells in the asthma group compared with the control group (203.5+/-84.6/mm2, mean+/-s.d. vs 37.7+/-8.7/mm2, P<0.05, n=4). In contrast, basophils were observed in the airway lumen, in the bronchial epithelium and in the submucosa. The number of basophils in the bronchioles was 81.8+/-55.5/mm2 (n = 4); however, basophils were not found at all in the airways of the control group. Although eosinophils, B lymphocytes and macrophages bear low affinity IgE receptors and could react with anti-IgE, the location of these cells in the close sections did not correspond closely with basophils. The presence of basophils in lung tissues obtained from fatal asthma patients supports the view that basophils play a role in the pathogenesis of bronchial asthma.
Adult↗
Activation of bone marrow eosinophils in asthma.
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Asthma↗
[Tumor recurrence and grade and/or stage progression in low grade tumor. Analysis of 154 patients with superficial transitional cell carcinoma of the bladder].
The clinicopathological findings of 154 patients with initially superficial (stage pTa or pT1) transitional cell carcinoma in the bladder were analyzed to study risk factors for tumor recurrence or progression in grade and/or stage of the low grade tumor. The number, size, grade and stage of tumor at the first presentation were the significant predictors for the first tumor recurrence. However, only the tumor grade and size at the first recurrence were the predictors for the second recurrence. In 19 (33%) of 57 patients with recurrence, low grade superficial tumor progressed in tumor grade and/or muscle invasion. The progression of low grade tumor correlated with short tumor free intervals and tumor size. Grade and/or stage progression was observed in 14% of the tumor of 1 cm or less in diameter, whereas in 42% of the tumor more than 1 cm in diameter (Chi-square test: p < 0.25). These results indicate that tumor grade and size are important predictors for recurrences. Short tumor free intervals and sizes of tumors in the second recurrence are significant predictors for grade and/or stage progression in low grade superficial cancer.
Adult↗
[Portal blood levels of 5-FU administered intraperitoneally].
Portal blood levels of 5-FU were measured after intraperitoneal administration of 15 mg/kg of 5-FU in two patients who had a bypass operation for unresectable pancreatic cancer. The portal concentration of 5-FU increased rapidly. Tmax was 8-11 minutes and Cmax was 29-31 micrograms/ml. The peripheral blood levels followed the portal levels. Also, AUC of the portal levels seemed to be higher than the peripheral levels. The direct instillation of 5-FU into the peritoneal cavity might offer an alternative means of achieving higher regional drug levels, such as intraperitoneal and intraportal levels, without enhanced systemic toxicity.
Fluorouracil↗
High incidence of p53 gene mutation in human ovarian cancer and its association with nuclear accumulation of p53 protein and tumor DNA aneuploidy.
Using the polymerase chain reaction and single-strand conformation polymorphism analysis, p53 gene mutations were examined in 24 cases of ovarian tumor including 14 ovarian carcinomas and 2 borderline cases of common epithelial type, 7 germ cell tumors, and one stromal tumor. Abnormal bands indicating mutations were detected in 12 (50%) of the cases examined, being present most frequently in common "epithelial" ovarian carcinoma (71%, 10/14). One case each of squamous cell carcinoma originating in a dermoid cyst and anaplastic dysgerminoma were positive for mutation. Direct sequencing confirmed 12 mutations and revealed G-->A and G-->C nucleotide changes in 5 and 3 cases (42% and 25%), respectively. The mutation was localized at the CpG site of the gene in 3 cases. Immunohistochemical examination of p53 protein in 21 cases and DNA flow-cytometrical analysis in 17 cases were also performed. Nuclear accumulation of the p53 protein and DNA aneuploidy pattern were detected in 11 (52%) and 9 (53%) cases, respectively. These were significantly correlated with p53 gene mutation (P < 0.01 and P < 0.05, respectively; Fisher's exact test). Neither mutation of the p53 gene, nuclear accumulation of p53 protein nor DNA aneuploidy was detected in borderline cases of common "epithelial" type, typical dysgerminoma and immature teratoma. These results suggest that p53 gene mutation, nuclear accumulation of the protein and the DNA aneuploidy pattern are events occurring almost simultaneously in the progression of ovarian tumors, and that p53 abnormalities seem to be correlated with a high grade of malignancy.
Aneuploidy↗
Enzymatic synthesis of 2-chloro-4-nitrophenyl 4,6-O-3-ketobutylidene beta-maltopentaoside, a substrate for alpha-amylase.
A transglycosylation reaction with 2-chloro-4-nitrophenyl beta-maltoside as an acceptor was done with 4,6-O-3-ketobutylidene maltopentaose and Bacillus macerans cyclodextrin glucanotransferase in an aqueous solution containing 50% n-propanol, and there were two main transglycosylation products. They were identified as 2-chloro-4-nitrophenyl 4,6-O-3-ketobutylidene beta-maltopentaoside and 2-chloro-4-nitrophenyl 4,6-O-3-ketobutylidene beta-maltohexaoside, and their yields were 30% and 21% respectively on the basis of the decrease of 4,6-O-3-ketobutylidene maltopentaose. For the production of 2-chloro-4-nitrophenyl 4,6-O-3-ketobutylidene beta-maltopentaoside at high substrates concentrations, the addition of n-propanol in this reaction not only increased the solubility of 2-chloro-4-nitrophenyl beta-maltoside sufficiently but also suppressed side reactions.
1-Propanol↗
[A case report: occurrence of bladder carcinoma in the defunctionalised bladder in a patient after operation of left ureteral carcinoma].
An eighty year-old woman was diagnosed as left ureteral carcinoma and right dysfunctional kidney in 1984. Left partial ureterectomy, partial cystectomy, and left ureterocutaneostomy were performed and her bladder was left behind after the operation. In 1990 (72 months after the operation), she was aware of bleeding from the urethra. A bladder wash was performed, demonstrating malignant cells on cytological examination, and cystoscopy revealed a papillary tumor in the defunctionalized bladder. Total cystectomy was performed and the histological findings of the surgical specimen showed transitional cell carcinoma, grade 2, pTaNoMo. This case shows the need for periodic examinations of the defunctionalized bladder, because renal pelvic and ureteral carcinoma tends to recur in the bladder within two years after operation and because tumors rarely developed in the defunctionalized bladder.
Adenocarcinoma↗
Rapid and sensitive method for erythropoietin determination in serum.
An enzyme-linked immunosorbent assay (ELISA) method for erythropoietin determination has been established by using two kinds of monoclonal antibodies with specific affinities to erythropoietin. Besides being rapid (2.5 h) and sensitive (detection limit 0.3 int. unit/L), the present method gives accurate results and is easy to perform. The method may be clinically applicable for discriminative diagnosis of polycythemia and analyses of various anemic conditions.
Anemia↗
Allelic loss on chromosome 17p and p53 mutations in human endometrial carcinoma of the uterus.
To understand the involvement of allelic losses and inactivation of tumor suppressor genes for the development of endometrial carcinoma of the uterus (EC), 24 cases of EC were examined for loss of heterozygosity (LOH) using a total of 57 polymorphic DNA markers covering all 23 pairs of chromosomes. LOH was observed at 27 loci on 10 different chromosomes, i.e., chromosomes 1, 3, 6, 11, 13, 15, 17, 18, 20, and 21, but was not detected at loci on chromosomes 4, 5, 7, 9, 10, 12, 14, 16, and X. It was observed only in seven of 24 cases, and the other 19 cases did not show LOH at any loci examined, including five cases of tumors with a high proportion of adenomatous hyperplasia. Among seven tumors with LOH at one or more loci, five tumors showed LOH at loci on the short arm of chromosome 17. Furthermore, mutations of the p53 gene, which is located on the short arm of chromosome 17, were detected in three of these 24 tumors by a polymerase chain reaction-single strand conformation polymorphism analysis and subsequent DNA sequencing. In two of these three tumors, p53 mutations were accompanied by the loss of wild-type p53 alleles. These results suggest that inactivation of the p53 gene is involved in the development of EC as in the case of several other types of human cancers.
Alleles↗
Determination of alpha-amylase using a new blocked substrate (3-ketobutylidene beta-2-chloro-4-nitrophenyl-maltopentaoside).
A new substrate, 3-ketobutylidene beta-2-chloro-4-nitrophenylmaltopentaoside (3KB-CNPG5), was used for the determination of alpha-amylase (EC 3.2.1.1) in serum and urine. Under this alpha-amylase assay condition, 3KB-CNPG5 is resistant to glucoamylase and alpha-glucosidase, which are auxiliary enzymes, because the 4- and 6-positions of the non-reducing-end glucose residue are modified by the 3-ketobutylidene group. The assay using 3KB-CNPG5 for alpha-amylase activity is a highly sensitive method that uses 2-chloro-4-nitrophenol (CNP) as an aglycone, and is a stable method for determination of alpha-amylase activity in biological fluids.
Chromatography, High Pressure Liquid↗
Structures of the asparagine-linked sugar chains of human chorionic gonadotropin from a patient with extragonadal germ cell tumour.
Human chorionic gonadotropin (hCG) purified from the urine of a male patient with extragonadal germ cell tumour contained four asparagine-linked sugar chains in one molecule. The sugar chains were quantitatively released from the polypeptide moiety by hydrazinolysis and recovered as oligosaccharides after N-acetylation. The oligosaccharide mixture was separated into a neutral (N) and three acidic (A1, A2 and A3) fractions by anion-exchange column chromatography. By sequential exoglycosidase digestion, methylation analysis and lectin column chromatography, the structures of these oligosaccharides were found to be the same as those of female gestational choriocarcinoma hCGs. Both contain eight kinds of sugar chains: triantennary, abnormal and normal biantennary, and monoantennary complex-type sugar chain with or without a fucosylated core portion.
Adult↗
Newly established uterine cervical carcinoma cell line with co-amplification of human papillomavirus DNA and c-myc gene.
A new human tumor cell line, NCC-c-CX-1 (CX-1), was established from a uterine cervical cancer xenografted in nude mice. This cell line harbored approximately 50 to 100 copies of human papillomavirus (HPV) type 18 DNA per haploid genome, and contained about 16-fold-amplified c-myc gene with rearrangement. These genomic alterations found in CX-1 cells were also present in both primary tumor and xenografted tumor. Histopathologically, original and xenografted tumors were poorly differentiated cancer and were characterized by neuroendocrine features such as positive neuron-specific enolase and chromogranin A by immunohistochemistry and abundant neurosecretory-type granules in the cytoplasm by electron microscopy. However, the established cell line had lost the neuroendocrine features. This cervical cancer cell line may be a useful model for studying cervical carcinogenesis, especially the interaction between HPV and c-myc oncogene.
Adult↗