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At least 649 records · Page 36Linked to original sources
PubMed2000

Dye- and gamma probe-guided sentinel lymph node biopsy in breast cancer patients: using patent blue dye and technetium-99m-labeled human serum albumin.

BACKGROUND: Sentinel lymph node (SLN) biopsy is a promising method for the diagnosis of the axillary nodal status. We examined the availability of the SLN biopsy using two mapping procedures: the dye- and gamma probe-guided method, and preoperative lymphoscintigraphy by gamma camera imaging. METHODS: We enrolled 48 patients with breast cancer. Technetium-99m-labeled human serum albumin was injected into the subdermal tissue above the primary tumor or biopsy cavity, and preoperative gamma camera imaging was performed. After induction of general anesthesia, patent blue dye was injected into the peritumoral area prior to the surgical procedure. A handheld gamma-detection probe was used to assist in SLN detection. Careful dissection was performed to identify blue-stained afferent lymphatic vessels and nodes. An SLN was defined as any blue and/or radioactive node, and was excised. After SLN biopsy, axillary lymph node dissection of level I, II, and III was completed, in order to confirm the diagnostic ability of the SLN biopsy. RESULTS: Intraoperative SLN identification of axillary lesions was successful in 43 of 48 patients (90%). The dye- and gamma probe-guided method was successful in 25 patients (52%), the dye-guided method alone succeeded in 11 patients (23%), and the gamma probe-guided method alone succeeded in 7 patients (15%). Preoperative lymphoscintigraphy revealed axillary focal accumulations in 29 of 48 patients (60%). All patients who underwent successful preoperative SLN identification by lymphoscintigraphy had successful intraoperative SLN identification. A diagnostic accuracy of 95%, a sensitivity of 89%, and a specificity of 100% were achieved in the diagnosis of axillary metastasis. Internal mammary SLNs were identified in four patients intraoperatively, but we could not detect cancer metastasis in the internal mammary SLNs. CONCLUSIONS: The dye-guided and gamma probe-guided methods were complementary. Preoperative lymphoscintigraphy was useful to predict intraoperative SLN identification. Further study is necessary to assess the role of SLN biopsy of the internal mammary lymph nodes.

Adult↗
PubMed2005

Removal of Supranol Yellow 4GL by adsorption onto Cr-intercalated montmorillonite.

Cr(III)-intercalated montmorillonite was utilized as an adsorbent for the removal of the organic pollutant, Supranol Yellow 4GL, a synthetic dye used for chemical fibres. The material was prepared by the reaction of Na montmorillonite with a base-hydrolyzed solution of Cr nitrate salt (OH(-)/Cr(3+) molar ratios of 2). XRD data showed that the interlayer spacing (d(001)) of montmorillonite was increased from 12.35 to 23.06 Angstroms. The kinetics and mechanism of the adsorption of the acid dye, Supranol Yellow 4GL, on Cr(III)-intercalated montmorillonite was investigated. The equilibrium time was reached within 30 min. The process follows pseudo-second-order rate kinetics. The Langmuir isotherm described the adsorption data over the concentration range (20-160 mg/l). The separator factor R(L) revealed the favourable nature of this adsorption process. Also, the thermodynamic parameters such as DeltaS degrees, DeltaH degrees, DeltaG degrees were determined.

Adsorption↗
PubMed2007

Blue-native gel electrophoresis for the characterization of protein complexes in plants.

High-resolution protein separation procedures are an important prerequisite for proteome analyses. Classically, protein separations are based on 2D IEF/SDS-PAGE. Unfortunately, this technique only poorly recovers hydrophobic proteins, and it is not compatible with analyses of proteins in native state. Blue-native PAGE represents a powerful alternative. It is based on the careful integration of negative charge into proteins and protein complexes by the anionic wool dye Coomassie blue, and it allows protein analyses under native ("blue-native") conditions. Upon combination with SDS-PAGE for a second gel dimension, protein complexes separated by the blue-native dimension are dissected into their subunits, which form vertical rows of spots on the resulting 2D gels. 2D blue-native/SDS-PAGE ideally complements 2D IEF/SDS-PAGE in proteomics.

Cell Fractionation↗
PubMed1985

Qualitative analysis of Coomassie-blue-stained proteins from normal prostate, benign prostatic hypertrophy, or adenocarcinoma of the prostate, separated by two-dimensional protein electrophoresis.

Samples from nine normal prostates (N), ten benign hypertrophic prostates (BPH), and eight adenocarcinomas of the prostate (ACP), obtained from the National Prostatic Cancer Project were analyzed by two-dimensional (2D)-protein electrophoresis. Qualitative analysis of Coomassie-blue-stained acidic proteins between pI 3.5 and 6.8 yielded these results: The total number of enumerated proteins was 136 (N), 107 (BPH), and 147 (ACP). Ninety-two denoted proteins were deemed common to the three kinds of samples, three were considered present in N and BPH, 7 in BPH and ACP, and 28 in N and ACP. Thirteen, five, and 20 proteins were detected only in N, BPH, or ACP. These proteins cannot be considered "unique" to their respective samples, since amounts of protein that do not exceed the minimum sensitivity of the dye may be present in the other samples. The similarity in the patterns of proteins from these three types of prostate samples is consistent with the following hypothesis: The distribution of proteins from histologically ambiguous prostate samples will correlate more closely with these protein profiles than with those associated with cancers originating from other organs.

Adenocarcinoma↗
PubMed1990

Quantitation of proteins bound to polyvinylidene difluoride membranes by elution of coomassie brilliant blue R-250.

A rapid and simple method for the quantitation of stained proteins bound to polyvinylidene difluoride (PVDF) membranes via the elution of Coomassie brilliant blue R-250 is described. A mixture of standard proteins was resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotted onto PVDF membranes. Spectrophotometric analysis of dye eluted from protein bands in the range of 0.5-10 micrograms gave a linear change in the absorbance at 595 nm. Maximal absorbance readings were attained following 5 min of dye elution, and the readings remained unchanged for elution times up to 60 min. The method requires no unusual reagents or equipment, is suitable for the analysis of multiple samples, and does not consume the protein in the process of quantitation. This technique provides a useful means for the quantitation of proteins bound to PVDF membranes prior to amino acid sequence determination, immunological analysis, or other biochemical characterizations.

1-Propanol↗
PubMed1986

Gold staining in cellulose acetate membranes.

A novel method for revealing proteins after electrophoresis on cellulose acetate is described, based on adsorption of mixed gold-Tween 20 micelles onto TCA-fixed protein bands. The sensitivity is 2-300 times higher than conventional Coomassie Blue staining and ca. 10 fold higher than silver coloring, detecting barely 1 ng protein/mm2 gel. Acidic and ammoniacal silver dyeing, based on ionic silver, perform very poorly on cellulose matrices, while a new variant, colloidal silver, stains protein bands on a clear background, but with a sensitivity ca. 10 times lower than micellar gold. Urines, cerebrospinal fluid and other biological liquids can be revealed without resorting to a concentration step prior to electrophoresis. The method is simple to perform and does not require any destaining step.

Blood Proteins↗
PubMedDate not supplied

The sentinel node biopsy is a new standard of care for patients with early breast cancer.

Sentinel lymph node biopsy (SLNB) is a simple technique which uses subdermal or peritumoral injection of vital blue dye and/or both substances together to identify the first lymph node(s) draining the primary tumor. It has been shown to accurately predict axillary node status in patients with clinically node negative breast cancer. The SLNB is emerging as a new standard of care in patients with early breast cancer. However, certain criteria should be fulfilled for its safe application.

Axilla↗
PubMedDate not supplied

Marginal leakage of Class II glass-ionomer-silver restorations, with and without posterior composite coverage: an in vitro study.

The aims of this in vitro investigation were: 1) to assess marginal leakage around three types of class II restorations, using: a) a composite resin with a glass ionomer lining; b) a glass ionomer cerment with composite resin coverage; c) a glass ionomer cermet without composite resin coverage and 2) to study the effect of thermocycling on marginal defects, by means of radiographs and SEM micrographs of the margins. Fifty-eight class II cavities were prepared in extracted primary molars. The teeth were divided into three groups and restored as follows: Group A--Ketac Bond (liner) and P-30; Group B--Ketac Silver and P-30(sandwich); Group C--Ketac Silver only. The restored teeth were thermocycled and marginal leakage was assessed from the degree of dye penetration on the sections. Dye penetration at the occlusal margins increased in the sequence A less than B less than C. The differences between group C and Groups A and B were statistically significant. Severe penetration of the dye was observed at the cervical margins with no statistically differences between the groups. Deterioration of margins due to thermocycling was observed for all groups, but these defects were not evident on the radiographs. Ketac Silver with and without composite coverage did not prevent marginal leakage when utilized in class II restorations in vitro.

Cermet Cements↗
PubMed1986

The annual cycle of Erinaceus europaeus L. as a model for a further study of cytochemical heterogeneity in Purkinje neuron nuclei.

The cytochemical heterogeneity of nuclei in Purkinje cells, which reflects differences in metabolic states within the population, has been studied by comparing cerebellar hemispheres of the hedgehog during periods of activity and hibernation. Cerebellar granule cells and hepatocytes served as controls for diploid cells. Three different cytochemical stains (Feulgen reaction, Propidium iodide and Hoechst 33342 fluorochromes) were used in order to evaluate by microdensitometry and microfluorometry how much the heterogeneity is dependent on the variations in DNA content and degree of chromatin compaction. In the active period, Feulgen-DNA values of Purkinje cells were more widely dispersed than in control cells. Some values (18-40% of the population) exceeded (hyperdiploid) the diploid value. With Propidium iodide and Hoechst 33342 (under conditions for quantitative evaluation of DNA), a few values also exceeded the maximum value for control cells. We conclude that there is nuclear heterogeneity (diploid-hyperdiploid) in the Purkinje neuron population during normal activity. The percentage of hyperdiploid nuclei changes according to the technique used; in particular, the comparison of the different stains indicates that a different chromatin compaction is mainly responsible for hyperdiploid Feulgen-DNA values, but in a few hyperdiploid cells there is an actual surplus of DNA. In view of the modalities of detection of DNA by the different cytochemical approaches utilized, this may be mainly single-stranded DNA. During hibernation nuclear heterogeneity is still present in Purkinje neurons, but with a shift of the distribution towards lower values; hyperdiploid nuclei were not detected with all the procedures used. Moreover, when Propidium iodide is used at the concentration suitable for qualitative studies, the dye intercalation is lower than during activity. Data suggest that both decreased DNA and increased chromatin compaction occur in Purkinje cells, thus pointing to a lower metabolic activity of these neurons in hibernating animals. The changes in diploid granule cells were always less than those in Purkinje neurons and could be ascribed to a further increase in chromatin compaction only, not requiring changes in DNA content.

Animals↗
PubMed2004

Sentinel lymph node detection in early cervical cancer with combination 99mTc phytate and patent blue.

OBJECTIVE: The purpose of this study was to examine sentinel lymph node (SLN) detection in patients with early stage cervical cancer using (99m)Tc phytate and patent blue dye and to compare our method with published findings utilizing other radioisotopic tracers. PATIENTS AND METHODS: A total of 20 consecutive patients with cervical cancer scheduled for radical hysterectomy and total pelvic lymphadenectomy at our hospital underwent SLN detection study. The day before surgery, lymphoscintigraphy was performed with injection of 99m-technetium ((99m)Tc)-labeled phytate into the uterine cervix. At surgery, patients underwent lymphatic mapping with a gamma-detecting probe and patent blue injected into the same points as the phytate solution. RESULTS: At least one positive node was detected in 18 patients (90%). A total of 46 sentinel nodes were detected (mean, 2.3; range, 1-5). Most sentinel nodes were in one of the following sites: external iliac (21 nodes), obturator (15 nodes), and parametrial (7 nodes). Eleven (24%) sentinel nodes were detected only through radioactivity and two (4%) were detected only with blue dye. The sensitivity, specificity, and negative predictive value for SLN detection were all 100%. Nine published studies involving 295 patients had a summarized detection rate of 85%. Summarized sensitivity, specificity, and negative predictive value were 93%, 100%, and 99%, respectively. CONCLUSION: Combination of (99m)Tc phytate and patent blue is effective in SLN detection in early stage cervical cancer.

Adult↗
PubMed2003

Sentinel node identification and the ability to detect metastatic tumor to inguinal lymph nodes in squamous cell cancer of the vulva.

OBJECTIVES: The goal of this study was to identify one or more inguinal sentinel nodes in patients with primary squamous cell carcinoma of the vulva and to determine the ability of the sentinel node to predict metastasis to the inguinal lymphatic basin. METHODS: Techniques employing technetium-99m (Tc-99m) sulfur colloid and isosulfan blue dye were utilized to identify sentinel nodes in the inguinal lymphatic beds. Technetium-99m sulfur colloid was injected intradermally at the tumor margins 90-180 min preoperatively followed by a similar injection of isosulfan blue dye 5-10 min before the groin dissection. A handheld collimated gamma counter was employed to identify Tc-99m-labeled sentinel nodes. Lymphatic tracts that had taken up blue dye and their corresponding sentinel node were also identified and retrieved. A completion inguinal dissection was then performed. Each sentinel node was labeled as hot and blue, hot and nonblue, or cold and blue. The sentinel nodes were subjected to pathologic examination with step sections and nonsentinel nodes were evaluated in the standard fashion. RESULTS: Twenty-one patients with a median age of 79 were entered onto protocol and a total of 31 inguinal node dissections were performed. A sentinel node was identified in 31/31 (100%) groin dissections with the use of Tc-99m. Isosulfan blue dye identified a sentinel node in 19/31 (61%) groin dissections. Surgical staging revealed 7 patients with stage I disease, 5 with stage II disease, 5 with stage III disease, and 4 with stage IV disease. Lymph nodes in 9 groin dissections were found to have metastatic disease, and in 4 of these dissections, the sentinel node was the only positive node. Lymph nodes in 22 groin dissections had no evidence of metastasis. No false-negative sentinel lymph nodes were obtained (sentinel node negative and a nonsentinel node positive). CONCLUSION: Tc-99m sulfur colloid is superior to isosulfan blue dye in the detection of sentinel nodes in inguinal dissections of patients with vulvar cancer. A sentinel node dissection utilizing Tc-99m alone can identify a sentinel node in all inguinal dissections. Pathologic examination with step sections has shown the sentinel node to be an accurate predictor of metastatic disease to the inguinal nodal chain.

Carcinoma, Squamous Cell↗
PubMed1994

Chromatography of human immunoglobulin G on immobilized drimarene rubine R/K-5BL. Study of mild, efficient elution procedures.

A range of substances were screened to find eluents for human immunoglobulin G (IgG) which are retained with a strong affinity by immobilized Drimarene Rubine R/K-5BL. The strong affinity of IgG for the dye is partly due to the presence of copper in the dye. This was suggested by the effect of substances able to make coordination bonds with metals that elute the IgG and also the effect of metal stripping from the immobilized dye. Several mobile phase conditions were found that allowed desorption of retained IgG on immobilized Drimarene Rubine R/K-5BL without using a protein denaturant. A procedure was also devised for separating IgG2 from other IgG subclasses using chromatography on immobilized Rubine R/K-5BL and column development with an AMP gradient.

Amines↗
PubMed1999

Subareolar versus peritumoral injection for location of the sentinel lymph node.

BACKGROUND: Sentinel lymph node (SLN) biopsy is fast becoming the standard for testing lymph node involvement in many institutions. However, questions remain as to the best method of injection. The authors hypothesized that a subareolar injection of material would drain to the same lymph node as a peritumoral injection, regardless of the location of the tumor. METHODS: To test this theory, 68 patients with 69 operable invasive breast carcinomas and clinically node-negative disease were enrolled in this single-institution Institutional Review Board-approved trial. Patients were injected with 1.0 mCi of technetium-99 sulfur colloid (unfiltered) in the subareolar area of the tumor-bearing breast. Each patient received an injection of 2 to 5 cc of isosulfan blue around the tumor. Radioactive SLNs were identified using a hand-held gamma detector probe. RESULTS: The average age of patients entered into this trial was 55.2 +/- 13.4 years. The average size of the tumors was 1.48 +/- 1.0 cm. Thirty-two percent of the patients had undergone previous excisional breast biopsies. Of the 69 lesions, 62 (89.9%) had SLNs located with the blue dye and 65 (94.2%) with the technetium. In four patients, the SLN was not located with either method. All blue SLNs were also radioactive. All located SLNs were in the axilla. Of the 62 patients in which the SLNs were located with both methods, an average of 1.5 +/- 0.7 SLNs were found per patient, of which 23.2% had metastatic disease. All four patients in which no SLN was located with either method had undergone prior excisional biopsies. CONCLUSIONS: The results of this study suggest that subareolar injection of technetium is as accurate as peritumoral injection of blue dye. Central injection is easy and avoids the necessity for image-guided injection of nonpalpable breast lesions. Finally, subareolar injection of technetium avoids the problem of overlap of the radioactive zone of diffusion of the injection site with the radioactive sentinel lymph node, particularly in medial and upper outer quadrant lesions.

Adult↗
PubMed1995

[Simultaneous assay of Ca(2+)-ATPase and Na+, K(+)-ATPase activities of hemolysate by malachite green colorimetric method].

Activities of Ca(2+)-ATPase and Na+, K(+)-ATPase of hemolysate were simultaneously determined by a colorimetric method with malachite green dye. In the presence of Ca2+, EGTA or ouabain, inorganic phosphate released from ATP by hypotonic saponin-treated red cell lysate was used to estimate the activities of Ca(2+)-ATPase and Na2+, K(+)-ATPase. The method is simple, sensitive and stable for at least 4 h. The effects of concentration of saponin and Ca2+, the storage of hemolysate, and the activator and inhibitor of the enzymes on activity were discussed.

Calcium-Transporting ATPases↗
PubMed2004

A prospective evaluation of radiocolloid and immunohistochemical staining in colon carcinoma lymphatic mapping.

BACKGROUND: Although the utility of lymphatic mapping (LM) and sentinel lymph node (SLN) biopsy in patients with melanoma and breast carcinoma has been well documented, this same is not true for patients with colon carcinoma. The authors previously reported a high false-negative rate for SLN biopsy in patients with colon carcinoma using isosulfan blue dye alone. The objective of the current study was to determine whether radiocolloid would increase the sensitivity of LM/SLN biopsy in patients with colon carcinoma. METHODS: The authors performed LM on 57 patients with colon carcinoma using both isosulfan blue dye and radiocolloid. The SLN(s) were identified by either their blue color or by increased radioactivity. The SLNs then underwent both routine histologic sectioning and immunohistochemical (IHC) staining for cytokeratins. RESULTS: An SLN was identified in 56 patients (98%). Radiocolloid was able to identify only 1 additional positive SLN (9%). Overall, it was found that the disease had metastasized to the lymph nodes in 22 patients, even though there was no evidence of disease in the SLN(s) in 11 of those 22 patients on routine histologic sectioning (false-negative rate, 50%; sensitivity, 50%). In five patients, IHC of the SLN was the only indicator of metastatic disease. The inclusion of IHC-positive SLNs in these calculations would decrease the false-negative rate to 17% and would increase the sensitivity of SLN biopsy to 83%. CONCLUSIONS: In the current study, the addition of radiocolloid did not increase the sensitivity of detection of positive SLN(s) compared with the use of isosulfan blue dye alone. IHC of the SLN potentially may increase the sensitivity of LM and reduce the false-negative rate. However, the long-term prognostic significance of IHC in patients with colon carcinoma remains controversial.

Adult↗
PubMed2001

Sentinel lymph node detection for breast cancer: which patients are best suited for the patent blue dye only method of identification?

BACKGROUND: The objectives of this study were, first, to define the preoperative criteria for using solely the blue dye method and, second, to decrease its operator dependence in predicting axillary lymph node status. METHODS: Two hundred fifty-three women consecutively identified with operable breast cancer underwent sentinel lymph node (SLN) detection by the patent blue dye method followed by completion axillary lymph node dissection. A standard pathological examination was performed for all SLN. Then, a pathological color quality assessment (PCQA), which checked for the presence of the blue dye, was performed on the paraffin blocks of the nonmetastatic SLN. Six preoperative identifiable variables likely to influence the detection rate were examined. RESULTS: The surgical detection (sd) rate was 84% (213 of 253) and the PCQA rate was 73% (185 of 253). Only breast size (sd, P = .0005; PCQA, P = .0007) and body mass index < or =30 (sd, P = .005; PCQA, P = .0007) were significant for SLN identification. Multivariate analysis revealed two independent factors influencing SLN identification: breast size (sd, P = .0001; PCQA, P = .002) and the timing of injection-injection prior to lumpectomy (sd, P = .04). CONCLUSIONS: The optimal patient features for identifying the SLN by the patent blue dye method are small or medium-sized breasts, low body fat, and that the procedure is carried out prior to tumor excision. The PCQA offers a useful second assessment of the surgically removed SLN, introducing an independent element of quality control.

Adult↗
PubMed1989

Sensitivity and variability of the Bradford protein assay in the presence of detergents.

The effects of Triton X-100, sodium dodecyl sulfate (SDS), and urea on the response of Coomassie blue G to 16 different proteins and peptides of Mr 1140 to 146,000 were studied to assess the significance of protein conformation and of ionic and nonionic interactions for the dye response to individual proteins. Triton X-100 at a final concentration of 0.008% (v/v) increased the sensitivity of the Bradford assay toward all proteins of Mr 5700 or higher by an average 33%. Increases ranged from +11% with myelin basic protein to +128% with aprotinin. The relative range of absorbance of proteins and deviations from bovine serum albumin decreased by approximately 25%. Triton X-100 appears to facilitate nonionic interactions of the dye with proteins of limited capacity for ionic binding. Conformation of proteins also seemed to be of some significance because the chaotropic agent urea (0.16 M final concentration) increased sensitivity of the assay by 14%. Sensitivity of the assay was lowered by SDS (0.004% final concentration, w/v) by an average 75% from that of the control assay. The results indicate that the incorporation of low concentrations of a nonionic detergent may be useful in improving sensitivity and variability of the Bradford assay.

Detergents↗
PubMed2001

Angle-resolved second-harmonic light scattering from colloidal particles.

We report angle-resolved second-harmonic generation (SHG) measurements from suspensions of centrosymmetric micron-size polystyrene spheres with surface-adsorbed dye (malachite green). The second-harmonic scattering profiles differ qualitatively from linear light scattering profiles of the same particles. We investigated these radiation patterns using several polarization configurations and particle diameters. We introduce a simple Rayleigh-Gans-Debye model to account for the SHG scattering anisotropy. The model compares favorably with our experimental data. Our measurements suggest scattering anisotropy may be used to isolate particle nonlinear optics from other bulk nonlinear optical effects in suspension.

Anisotropy↗