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PubMed1999

[Identification of sentinel lymph node in breast cancer: experience from the Institut Curie].

Sentinel lymph node (SN) biopsy is a recently developed, minimally invasive technique for staging the axilla in breast cancer. This new procedure of selective lymphadenectomy has been the subject of several studies, but there is not currently a consensus of opinion to define which is the best method of identification. At the Institut Curie since 1996, we have been using the Patent blue dye. The current series present the result of 122 patients with T1, T2, N0 or N1a breast cancer consecutively operated between december 1997 and august 1998. Sentinel nodes were identified in 107 out of 122 (87.7%) and accurately predicted axillary nodal status in 104 out of 107 (97.1%) cases. Three out of 35 node positive patients would have been missed with sentinel node biopsy alone, for a false negative rate of 8.5%. In all 3 cases, one lymph node presented with a micrometastases. In 15 cases out of 35 with metastatic axillary nodes, the only positive node was the SN (43%). The encouraging results of this study shows that it is possible to identify, in a large number of cases, the sentinel node by means of Patent blue dye only. This article detailed the technique used and reviews the literature concerning other methods of identification.

Adult↗
PubMed2006

Sentinel node detection with the blue dye technique in early cervical cancer.

OBJECTIVE: The aim of this study was to investigate the feasibility of sentinel node detection with the blue dye technique in early cervical cancer. METHODS: In a retrospective study conducted between January 2000 and February 2005, 47 women with early cervical cancer (6 patients FIGO Stage I A, 38 patients FIGO Stage I B, 2 patients FIGO Stage II A, 1 patient FIGO Stage II B) who underwent class II-III radical hysterectomy with pelvic lymphadenectomy were identified. Prior to surgery 1 ml of blue dye (lymphazurin 1%) was injected into the four quadrants of the cervix. RESULTS: The detection rate for sentinel nodes was 83% (39/47 patients). The median number of sentinel lymph nodes per patient was two. Nine patients had positive sentinel nodes. In one patient the sentinel lymph node procedure revealed to be false-negative. Positive predictive value and specificity were both 100%. The sensitivity and negative predictive value were 90% and 97%, respectively. CONCLUSIONS: Sentinel node detection has become a main field of interest in gynecological oncology. Our detection rate and sensitivity rate using the blue dye technique in cervical cancer are comparable to those in previously published data. However, recent data on a combined radioactively labeled albumin and blue dye technique show even more promising results. The clinical validity of the combined techniques must be evaluated prospectively in larger studies.

Adenocarcinoma↗
PubMed1994

[Protein determination by binding with the dye Coomassie brilliant blue G-250].

Possible applications of the Coomassie brilliant blue G-250 dye method for protein assay are reviewed. The physico-chemical and optical properties of the dye are considered in detail. The data on the protein-binding mechanism are generalized. Modifications of the Coomassie blue G-250 quantitative protein assays are critically considered. The applications of the method in medicine, biochemistry and biotechnology are discussed.

Animals↗
PubMed2006

Investigation of polymerisation shrinkage strain, associated cuspal movement and microleakage of MOD cavities restored incrementally with resin-based composite using an LED light curing unit.

OBJECTIVES: To investigate the polymerisation shrinkage strain, associated cuspal movement, degree of conversion (DC) and cervical gingival microleakage of mesio-occlusal-distal (MOD) cavities restored with four resin-based composite (RBC) filling materials placed incrementally using a light emitting diode (LED) light curing unit (LCU). METHODS: Standardised extensive MOD cavity preparations on extracted teeth were performed on 40 sound upper premolar teeth. Restoration of the teeth involved the placement of RBCs in eight increments with the appropriate bonding system before irradiation using an LED LCU. Buccal and palatal cusp deflections at each stage of polymerisation were recorded using a twin channel deflection measuring gauge. Following restoration, the teeth were thermocycled, immersed in a 0.2% basic fuchsin dye for 24 h, sagittally sectioned and examined for cervical microleakage. The DC was determined using a Fourier transform infra-red (FT-IR) spectrometer. RESULTS: No significantly difference (P=0.677) in cuspal movement was recorded for Z100 (13.1+/-3.2 microm) compared with Filtek Z250 (8.4+/-3.5 microm), P60 (7.3+/-3.8 microm) and Admira (6.7+/-2.7 microm). The LED LCU deflections were compared with a halogen LCU used in a conventional (Fleming GJP, Hall D, Shorthall ACC, Burke FJT. Cuspal movement and microleakage in premolar teeth restored with posterior filling materials of varying reported volumetric shrinkage values. Journal of Dentistry, 2005;33:139-146) and soft-start mode (Fleming GJP, Cara RR, Palin WM, Burke FJT. Cuspal movement and microleakage in premolar teeth restored with posterior filling materials cured using 'soft-start' polymerization. Dental Materials, 2006, , in press) and a significant reduction in cuspal movement was identified for curing type and material type (P<0.001 and P=0.002, respectively). No significant differences were noted between the four RBC materials investigated when the DC or microleakage scores were examined for the LED LCU. SIGNIFICANCE: It would appear that irradiation of RBCs using the LED LCU offered a significant reduction in associated cuspal movement in large MOD cavities. However, the microleakage scores following polymerisation were significantly increased with dye penetration into the pulp chamber from the axial wall evident in teeth restored with the LED LCU.

Bicuspid↗
PubMed2001

Self-oligomerization and protein aggregation of alpha-synuclein in the presence of Coomassie Brilliant Blue.

alpha-Synuclein has been implicated in various neurodegenerative disorders, including Parkinson's and Alzheimer's diseases, by its participation in abnormal protein depositions. As the protein has been suggested to play a significant role in the formation of the deposits which might be responsible for neurodegeneration, there is a strong demand to screen for alpha-synuclein-interactive small molecules. In this report, Coomassie Brilliant Blue (CBB) interaction of alpha-synuclein has been investigated with respect to induction of protein self-oligomerization in the presence of the chemical coupling reagent N-(ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline. Both CBB-G and CBB-R, which differ by only two methyl groups, induced the self-oligomerization of alpha-synuclein in a biphasic manner with optimal dye concentrations of 250 microM and 150 microM, respectively. The protein aggregates of alpha-synuclein induced by the dyes in the absence of the coupling reagent were analysed by electron microscopy. Whereas CBB-G induced formation of protein aggregates with a worm-like structure, CBB-R induced clear fibrilization of alpha-synuclein on a background of granular structures. CBB-R interacted with alpha-synuclein approximately twice as effectively as CBB-G (dissociation constants 0.63 microM and 1.37 microM, respectively). These dye interactions were independent from the acidic C-terminus of alpha-synuclein, which was reminiscent of the Alphabeta25-35 interaction of alpha-synuclein. However, the metal-catalysed oxidative self-oligomerization of alpha-synuclein in the presence of Cu2+/H2O2, which was augmented synergistically by Alphabeta25-35, was not affected by the dyes. This indicates that the dye binding site is also distinctive from the Alphabeta25-35 interaction site on alpha-synuclein. These biochemically specific interactions between alpha-synuclein and the dyes indicate that alpha-synuclein-interactive small molecules could provide a tool with which to approach development of diagnostic, preventive, or therapeutic strategies for various alpha-synuclein-related neurodegenerative disorders.

Binding Sites↗
PubMed1994

Chromophore-assisted laser inactivation of proteins is mediated by the photogeneration of free radicals.

Chromophore-assisted laser inactivation (CALI) is a technique that selectively inactivates proteins of interest to elucidate their in vivo functions. This method has application to a wide array of biological questions and an understanding of its mechanism is required for its judicious application. We report here that CALI is not mediated by photoinduced thermal denaturation but by photogenerated free radicals. Thermal diffusion calculations suggest that the temperature changes resulting from CALI are too small to cause thermal denaturation, and Arrhenius plots of CALI are inconsistent with a photothermal mechanism. CALI shows an energy dose reciprocity above a threshold and can be inhibited by free-radical quenchers, thus demonstrating a photochemical mechanism of protein inactivation. The type of quenchers that are effective in inhibiting CALI indicates that the active species is a hydrogen abstractor which is not derived from molecular oxygen. We suggest that the active free-radical species is the hydroxyl radical and its very short lifetime explains the spatial specificity of CALI such that half-maximal damage is effected within 15 A from the dye moiety and no significant damage occurs at 34 A. The data are consistent with free-radical formation resulting from a sequential two-photon process.

Coloring Agents↗
PubMed1980

Cytochemical properties of the conventional Schiff reagent fortified with Tris-buffer.

This paper presents a cytochemical study of a modified Schiff reagent in which 250 Tris-buffer has been incorporated in addition to 5 ml of N HCL and 1.0 g of potassium metabisulphite per 100 ml of an aqueous solution of 0.5% basic fuchsin. The results indicate production of an orange-coloured dye-reagent which when used on acid hydrolysed tissue sections reveals brilliantly stained nuclei. The shelf-life of this modified Tris-Schiff reagent is also much more than that of the conventional Schiff reagent. The in vitro absorption spectra of the orange-coloured dye-reagent together with those of the modified Tris-Schiff reagent with formalin as well as the conventional Schiff reagent with formalin are also presented. Apossible explanation for the increased shelf-life has been offered.

Animals↗
PubMed2003

Identification of sentinel lymph nodes in colon cancer depends on the amount of dye injected relative to tumor size.

Recent studies have shown that the sentinel lymph node (SLN) procedure might improve staging in colon cancer. However, low SLN identification and high false negative rates have also been reported. In a two-institution study, the SLN procedure with isosulfan blue 1% was performed according to a standardized protocol in 31 patients with open resections for colon cancer. Data were collected prospectively. The database was analyzed retrospectively to determine factors contributing to a low identification rate. The SLN identification rate was 87% and the false negative rate was 50%. Successful SLN identification was significantly associated with application of higher volumes of dye relative to the tumor diameter ( p = 0.04) and more frequent tumor localization in the sigmoid colon ( p = 0.04) as compared to missing SLN identification. The tumor diameter was not significantly different in the two groups. Sentinel lymph node identification in colon cancer depends on the amount of dye injected relative to the tumor size. Application of only 1 ml of dye-the amount generally recommended in the literature-is not sufficient in large tumors.

Aged↗
PubMed2005

A colorimetric assay for the determination of 4-diphosphocytidyl-2-C-methyl-D-erythritol 4-phosphate synthase activity.

A new method for the determination of the activity of 4-diphosphocytidyl-2-C-methyl-D-erythritol 4-phosphate synthase, the enzyme catalyzing the third reaction of the 2-C-methyl-D-erythritol 4-phosphate pathway for biosynthesis of isoprenoids, is described. This is an end-point assay based on the transformation of inorganic pyrophosphate, one of the products of the reaction, to phosphate by using inorganic pyrophosphatase as auxiliary enzyme. The phosphate formed is reacted then with the dye malachite green to yield a colored product which can be determined spectrophotometrically. The method is easy to perform, sensitive, and robust and can be used in automated high-throughput screening analyses for the search of inhibitors of the enzyme.

Colorimetry↗
PubMed1979

Model studies on the acriflavine-Feulgen reaction.

The specificity and quantitative reliability of the Feulgen-acriflavine-SO2 procedure was tested on polyacrylamide model films containing DNA. Noncovalent binding of acriflavine to DNA was observed when the washing procedure, as used in the classical way, was applied. The noncovalently bound acriflavine could be removed with an extra wash in acid-ethanol. The presence of SO2 in the staining solution has been found to enhance covalent binding significantly. The absorbance of films stained by our Feulgen-acriflavine-SO2 procedure is directly proportional to that obtained by the classical Feulgen-pararosanilline-SO2 procedure. The acriflavine-Feulgen procedure has also been tested using a commercial and a purified dye. The use of purified acriflavine, compared to a commercial sample did not result in a significant difference in the maximum absorbance value of stained DNA nor in the absorption or the fluorescence emission spectra of acriflavin covalently bound to DNA.

Acriflavine↗
PubMed2002

Sentinel node biopsy as an indicator for pelvic nodes dissection in early stage cervical cancer.

The purpose of this study was to investigate the feasibility of sentinel node frozen biopsy to minimize the extensive pelvic lymph nodes dissection in early stage cervical cancer patients on the basis that the risk of skip metastasis to the paraaortic area is negligible. Twenty-six patients with early stage cervical cancer were enrolled in this study. Technetium-99m colloid albumin (Tc(99m)) was injected intradermally around the tumor for allowing preoperative lymphoscintigraphy and intraoperative hand-held gama probe detection of seninel nodes. For visual detection, isosulfan blue dye was injected into the peritumoral sites before peritoneal opening. Postoperative morbidity and negative predictive value were the endpoints of this study. The 26 patients, ranging in age from 32 to 71 yr, underwent intraoperative sentinel nodes mapping. All the patients underwent complete pelvic lymph nodes dissection including para-aortic nodes. There was one case with positive non-sentinel nodes despite the negative sentinel node by frozen biopsy (negative predictive value, 95.2%). This new technique of sentinel node mapping is safe and simple to perform. Further clinical trials using the combination of Tc(99m) and isosulfan blue dye are warranted and this technique will make a true advance for less aggressive management of patients with early stage cervical cancer.

Adult↗
PubMedDate not supplied

Penetrability of dentinal tubules in adhesive-lined cavity walls.

This study investigated the penetrability of dentinal tubules in cavity walls lined with different dentin bonding systems. Occlusal Class I cavities were prepared in 93 premolars. The cavities in the control group had an intact smear layer without a lining, while those in the experimental group were lined with Gluma CPS, Scotchbond Multi-Purpose Plus or One-Step. The penetrability of the dentinal tubules was tested with a dye (basic fuchsin) or bacteria (S faecalis) immediately after adhesive lining and after one-month storage in water at 37 degrees C. Some of the lined samples were sectioned and examined under the SEM. In some samples in the experimental group, the dye penetrated to the pulp and bacteria up to 125 microm into the dentinal tubules immediately after lining. The Kruskal Wallis ANOVA and Tukey test showed the depth of dye and bacterial penetration to be significantly less in teeth with bonding systems than those in the control group (p<0.05). However, there was no statistically significant difference between the control and experimental groups after storage in water (p>0.05). SEM examination showed that the hybrid layer and resin tags were present in the cavity walls immediately after lining but absent after storage in water. Therefore, adhesive linings under the experimental conditions were ineffective in preventing dye or bacterial penetration of the dentinal tubules.

Analysis of Variance↗
PubMed1988

A stable colorimetric assay to measure toxin elevation of inorganic phosphate in bile.

A modified assay for the measurement of nanomole amounts of Pi in 50 microliters of bile is described. The assay is based on the formation of a complex between malachite green dye and phosphomolybdate under acidic conditions. Only three simple steps are required to produce a colored complex which remains stable for at least 3 h; these steps are precipitation with acid, addition of color reagent containing dye and surfactant, and incubation at 37 degrees C. Analysis of bile samples collected from a rat given the toxin 1,1-dichloroethylene demonstrated that the assay is well suited to routine assays of biliary Pi as an endogenous indicator of aberrant hepatobiliary function.

Animals↗
PubMed1992

Quantitation of proteins separated in N, N'-1,2-dihydroxyethylenebisacrylamide-crosslinked polyacrylamide gels.

A simple and rapid method for the quantitation of proteins separated either by sodium dodecyl sulfate-electrophoresis or by isoelectric focusing in slab gels is presented. The method is based on the solubility of polyacrylamide gels crosslinked with N, N'-1, 2-dihydroxyethylenebisacrylamide (DHEBA) in periodic acid. After electrophoretic separation proteins are stained with Coomassie brilliant blue G-250. DHEBA gels show considerable swelling during the staining and destaining process but can be shrunk to their normal size in a 10% (w/v) solution of ammonium sulfate. Stained bands are cut from the gel and solubilized in periodic acid. During dissolution the dye decolorizes. Protein concentration in the solution is determined by a modified Coomassie dye-binding assay. Quantitation is linear in the range of 100 ng to 5 micrograms and not disturbed by dissolved gel. Separations in N, N'-1, 2-dihydroxyethylenebisacrylamide-crosslinked gels show qualities similar to those in normal crosslinked gels.

Acrylamides↗
PubMed2002

Activated parthenium carbon as an adsorbent for the removal of dyes and heavy metal ions from aqueous solution.

Parthenium hysterophorous (L) is a perennial weed distributed all over the country. Carbonized parthenium activated with conc. H2SO4 and ammonium persulphate was effective in the removal of dyes, heavy metals and phenols. Variation in the percentage removal of adsorbates was observed with increase in the contact time. Among the adsorbates tested, the affinity of the activated parthenium carbon was highest for Hg2+, Methylene Blue and Malachite Green.

Adsorption↗
PubMed2001

Use of (99m)Tc-labeled liposomes encapsulating blue dye for identification of the sentinel lymph node.

UNLABELLED: Colloidal radiopharmaceuticals are commonly used in combination with blue dye for localization of the sentinel node. Liposomes are colloidal particles composed of spontaneously forming lipid spheres that can carry a wide variety of diagnostic and therapeutic agents. Conventional liposomes are poorly retained in lymph nodes (<2% of the subcutaneously injected dose). We have previously described a system for increasing the retention of liposomes in the lymph nodes by approximately 7-fold. This system is comprised of subcutaneously injected biotin-coated liposomes, followed by an adjacent injection of avidin. When the avidin moves into the lymphatic vessels, it causes aggregation of the biotin-coated liposomes that are also in the process of migrating through the lymphatic vessels. These aggregated liposomes become entrapped in the next encountered lymph node. In this study, we use this novel lymph node delivery system with liposomes that encapsulate blue dye, resulting in intense blue staining of the sentinel node. These liposomes can also be labeled with (99m)Tc, permitting scintigraphic imaging and radioguided probe localization of the sentinel node. METHODS: Liposomes coated with biotin and coencapsulating blue dye and glutathione were labeled with (99m)Tc using hexamethylpropyleneamine oxime. Rabbits were subcutaneously administered 0.3 mL (99m)Tc-biotin-liposomes containing blue dye in both hind feet, followed by a subcutaneous injection (0.3 mL) of 5 mg avidin in only one hind foot (experimental). The other hind foot served as a control. RESULTS: Labeling efficiencies (mean +/- SEM) for liposomes encapsulating blue dye were 92.1% +/- 1.9%. Necropsy at 24 h revealed that the popliteal node on the experimental leg receiving the avidin was intensely blue stained compared with virtually no blue coloration of the control node. Tissue counts of these nodes were 12.2 +/- 1.5 percentage injected dose (%ID) in the experimental node compared with 1.2 +/- 0.1 %ID in the control nodes (P<0.0001). CONCLUSION: Biotin-liposomes encapsulating blue dye can be successfully labeled with (99m)Tc, providing a convenient option for the visualization and radiolocalization of the sentinel node. This biotin-liposome/avidin system may also have potential for the delivery of therapeutic drugs and radiopharmaceuticals to lymph nodes.

Animals↗
PubMedDate not supplied

Dyes for caries detection influence sound dentin bond strength.

This study evaluated the influence of caries-detection dyes on the in vitro tensile bond strength of adhesive materials to sound dentin. Caries-free human molars were ground to expose superficial dentin. Two dyes (a commercial 0.5% basic fuchsin in propylene glycol and Cari-D-Tect) were applied to sound dentin and rinsed. Subsequently, the dentin was etched with phosphoric acid (35%) and rinsed, leaving a moist dentin surface. The adhesive (Prime & Bond 2.0) was applied in two layers and light cured. A composite (TPH Spectrum), a compomer (Dyract), and a hybrid ionomer (Advance) were used to prepare the bond-strength specimens with a 3-mm-in-diameter bonding area. Control groups were made without use of dyes. Six specimens were prepared for each group. After 24 hours in distilled water, tensile bond strength (MPa) was measured using a testing machine. Analysis of variance was used to evaluate the data. Without dyes, bond strengths of TPH Spectrum and Dyract with Prime and Bond 2.0 were similar and both values were significantly (P < 0.05) higher than that of Advance with Prime & Bond 2.0. Dyes for caries detection reduced the bond strength of TPH Spectrum and Dyract but not Advance when used with Prime and Bond 2.0.

Coloring Agents↗
PubMed1993

An improved Bradford protein assay for collagen proteins.

A modification of the protein determination method of Bradford adapted for collagen-rich samples is described. The use of Coomassie-based protein determination methods is limited by the great variation in colour yield obtained for different proteins. This is especially important in samples containing significant amounts of collagen where direct application of the methods of Lowry and Bradford results in underestimated values. Addition of small amounts of sodium dodecyl sulphate (SDS) (0.0035%) to the diluted solutions of Coomassie Brilliant Blue G used as dye reagent in the Bradford colorimetric assay caused a 4-fold increase in the colour response of three collagen proteins (Col I, III and IV) and a decrease in absorbance for various non-collagen proteins. The presence of SDS in the reagent did not result in a significant metachromatic shift of the collagen-dye complexes. This simple modification in the preparation of the reagent for the Bradford assay allows similar response curves to be obtained for collagen and non-collagen proteins, making the modified assay of potential use for protein determination in collagen-rich samples such as pancreatic extracts.

Collagen↗