Initial experience with sentinel node biopsy in breast cancer at the National Cancer Center Hospital East.
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BACKGROUND: Axillary lymph node dissection is an important procedure in the surgical treatment of breast cancer. Axillary lymph node dissection is still performed in over half of breast cancer patients having histologically negative nodes, regardless of the morbidity in terms of axillary pain, numbness and lymphedema. The first regional lymph nodes draining a primary tumor are the sentinel lymph nodes. Sentinel node biopsy is a promising surgical technique for predicting histological findings in the remaining axillary lymph nodes, especially in patients with clinically node-negative breast cancer, and a worldwide feasibility study is currently in progress. METHODS: Intraoperative lymphatic mapping and sentinel node biopsy were performed in the axilla by subcutaneous injection of blue dye (indigocarmine) in 88 cases of stage 0-IIIB breast cancer. Sentinel lymph nodes were identified by detecting blue-staining lymph nodes or dye-filled lymphatic tracts after total or partial mastectomy. Finally, axillary lymph node dissection was performed up to Levels I and II or more. RESULTS: Sentinel lymph nodes were successfully identified in 65 of the 88 cases (74%). In the final histological examination, the sentinel lymph nodes in 40 cases were negative, including four cases with non-sentinel-node-positive breast cancer (specificity, 100%; sensitivity, 86%). In nine (31%) of the 29 cases with histologically node-positive breast cancer, the sentinel lymph nodes were the only lymph nodes affected. Axillary lymph node status was accurately predicted in 61 (94%) of the 65 cases. CONCLUSIONS: Although it was the initial experience at the National Cancer Center Hospital East, sentinel node biopsy proved feasible and successful. This method may be a reasonable alternative to the standard axillary lymph node dissection in patients with early breast cancer. (+info)
An indigo-reducing moderate thermophile from a woad vat, Clostridium isatidis sp. nov.
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A Gram-positive, anaerobic, moderate thermophile, strain Wv6T, capable of reducing indigo dye, was isolated from a fermenting woad vat prepared essentially as in medieval Europe. Strain Wv6T formed rod-shaped cells, which occurred singly, in pairs or in chains and produced terminal oval endospores. Strain Wv6T was saccharolytic. Growth occurred at pH 5.9-9.9 (initial pH) with an optimum at 50 degrees C of pH 7.2 +/- 0.2 (constant pH). At pH 7.8, the temperature range for growth was 30-55 degrees C with the optimum at 49-52 degrees C. Comparative 16S rRNA gene sequence analysis demonstrated that the bacterium represents a hitherto unknown subline within rRNA cluster I Clostridium. Based on the results of the phylogenetic analysis and phenotypic criteria, it is proposed that the unknown moderate thermophile should be classified as Clostridium isatidis sp. nov., a new species of the genus Clostridium. The type strain of Clostridium isatidis is strain Wv6T (= NCFB 3071T). (+info)
Substrate specificity of naphthalene dioxygenase: effect of specific amino acids at the active site of the enzyme.
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The three-component naphthalene dioxygenase (NDO) enzyme system carries out the first step in the aerobic degradation of naphthalene by Pseudomonas sp. strain NCIB 9816-4. The three-dimensional structure of NDO revealed that several of the amino acids at the active site of the oxygenase are hydrophobic, which is consistent with the enzyme's preference for aromatic hydrocarbon substrates. Although NDO catalyzes cis-dihydroxylation of a wide range of substrates, it is highly regio- and enantioselective. Site-directed mutagenesis was used to determine the contributions of several active-site residues to these aspects of catalysis. Amino acid substitutions at Asn-201, Phe-202, Val-260, Trp-316, Thr-351, Trp-358, and Met-366 had little or no effect on product formation with naphthalene or biphenyl as substrates and had slight but significant effects on product formation from phenanthrene. Amino acid substitutions at Phe-352 resulted in the formation of cis-naphthalene dihydrodiol with altered stereochemistry [92 to 96% (+)-1R,2S], compared to the enantiomerically pure [>99% (+)-1R,2S] product formed by the wild-type enzyme. Substitutions at position 352 changed the site of oxidation of biphenyl and phenanthrene. Substitution of alanine for Asp-362, a ligand to the active-site iron, resulted in a completely inactive enzyme. (+info)
Effects of methylene blue, indigo carmine solution and autologous erythrocyte suspension on formation of adhesions after injection into rats.
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The aim of this study was to determine whether autologous erythrocyte suspension can be used as a dye for evaluation of tubal patency and whether it has any advantages over methylene blue or indigo carmine solutions. Reproductively healthy female nulliparous Wistar Albino rats (n = 30), aged 6 months, mass 165-195 g, were assigned randomly to three groups. Rats received a 1 ml i.p. injection of 5% (w/v) methylene blue solution (methylene blue group: n = 10), 5% (w/v) indigo carmine solution (indigo carmine group: n = 10) or 5% (v/v) fresh autologous erythrocyte suspension (autologous erythrocyte group: n = 10). At 4 weeks after injection, a small sterile opening was made in the peritoneal cavity of each rat. The cavity was rinsed once with TCM-199 to collect macrophages. The rinsed peritoneal contents were cultured overnight to evaluate macrophage activation. The peritoneal opening was expanded for evaluation of adhesion formation. Only one rat from the autologous erythrocyte group had intra-peritoneal adhesions (score 2), whereas all rats in the methylene blue group (score 1: n = 1; score 2: n = 4; score 3: n = 4; and score 4: n = 1) and seven rats in the indigo carmine group (score 1: n = 1; score 2: n = 2; score 3: n = 3; and score 4: n = 1) had intra-abdominal adhesions. Macrophage activity was observed in the cultured peritoneal contents collected from the methylene blue and indigo carmine groups but not from the autologous erythrocyte group. Adhesion formation could be due to macrophage activation caused by methylene blue and indigo carmine solutions. These results indicate that tubal patency can be observed by laparoscopy using autologous erythrocyte suspension. The results of this study are believed to be the first to indicate that a patient's own erythrocyte suspension could be used during observation of tubal patency by laparoscopy. However, further studies are required. (+info)
Orcein-picroindigocarmine--a new multiple stain.
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A new "orcein-picroindigocarmine staining", a colour combination of orcein, indigo carmine, and picric acid, was developed for histological applications. The new technique was tested on different human tissues. Colours ranging from red to brown, yellow, green and blue were observed in paraffine sections of tissues stained by this method. Nuclear structures in all tissues were stained dark brown to dark blue. Squamous epithelium was stained light brown with varying shades of blue in upper horny layers, whereas the ciliated epithelium was tinged blue grey. When connective tissue was stained, collagen fibrils appeared strongly blue next to elastic fibres, which took on a rust brown tinge; cellular components were all coloured brown. The matrix of hyaline cartilage was stained in different shades of blue, with the chondrocytes rust brown. Sections of bone components appeared dark blue to dark green. Skeletal muscle cells were coloured yellow and green with blue collagenous septa. The new staining is useful for distinguishing connective tissue components such as elastic fibres and collagen fibrils. It also demonstrates chondrocytes in favourable contrast to the cartilage matrix. The technique produces aesthetic staining colouring that could supplement histological investigations and provide an alternative to other staining materials. (+info)
Purification, stability, and mineralization of 3-hydroxy-2- formylbenzothiophene, a metabolite of dibenzothiophene.
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3-Hydroxy-2-formylbenzothiophene (HFBT) is a metabolite found in many bacterial cultures that degrade dibenzothiophene (DBT) via the Kodama pathway. The fate of HFBT in cultures and in the environment is unknown. In this study, HFBT was produced by a DBT-degrading bacterium and purified by sublimation. When stored in organic solvent or as a crystal, the HFBT slowly decomposed, yielding colored products. Two of these were identified as thioindigo and cis-thioindigo. The supernatant of the DBT-degrading culture contained thioindigo, which has not been reported previously as a product of DBT biodegradation. In mineral salts medium, HFBT was sufficiently stable to allow biodegradation studies with a mixed microbial culture over a 3- to 4-week period. High-performance liquid chromatography analyses showed that HFBT was removed from the medium. 2-Mercaptophenylglyoxalate, detected as benzothiophene-2,3-dione, was found in an HFBT-degrading mixed culture, and the former appears to be a metabolite of HFBT. This mixed culture also mineralized HFBT to CO2. (+info)
Cloning and expression of a Ralstonia eutropha HF39 gene mediating indigo formation in Escherichia coli.
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On complex medium Escherichia coli strains carrying hybrid plasmid pBEC/EE:11.0, pSKBEC/BE:9.0, pSKBEC/PP:3.3, or pSKBEC/PP:2.4 harboring genomic DNA of Ralstonia eutropha HF39 produced a blue pigment characterized as indigo by several chemical and spectroscopic methods. A 1,251-bp open reading frame (bec) was cloned and sequenced. The deduced amino acid sequence of bec showed only weak similarities to short-chain acyl-coenzyme A dehydrogenases, and the gene product catalyzed formation of indoxyl, a reactive preliminary stage for production of indigo. (+info)
Indirubin and indigo are potent aryl hydrocarbon receptor ligands present in human urine.
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Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates genes involved in xenobiotic metabolism, cellular proliferation, and differentiation. Numerous xenobiotic and biological compounds are known to interact with AhR, but it remains an orphan receptor, because its physiological ligand is unknown. We identified AhR ligands in human urine using a yeast AhR signaling assay and then characterized their properties. Two ligands, indirubin and indigo, were both present at average concentrations of approximately 0.2 nm in the urine of normal donors. Indirubin was also detected in fetal bovine serum and contributed half of the total AhR ligand activity. The activities of indirubin and indigo were comparable with or more potent than that of the archetypal ligand, 2,3,7,8-tetrachlorodibenzo-p-dioxin, in yeast AhR activation assays. We suggest that the endogenous levels and potencies of indirubin and indigo are such that they activate AhR-mediated signaling mechanisms in vivo. (+info)