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PubMed1998

En bloc staining of bone under load does not improve dye diffusion into microcracks.

Microdamage accumulation in bone has been implicated in the pathogenesis of some bone fractures, and in implant loosening. Standard techniques for staining microcracks may not allow all cracks to be stained. We tested the hypothesis that crack closure in bone cortices after removal of a bending load may prevent diffusion of stain to sites of microcrack nucleation. Following cyclic loading, 26 canine femurs were divided into a group stained en bloc while applying a four point bending load, and another group stained without an applied load. No differences in number or length of microcracks were observed, indicating that crack closure does not prevent diffusion of stain to the crack location. Staining under load is unnecessary.

Animals↗
PubMed2006

Enhanced malachite green removal from aqueous solution by citric acid modified rice straw.

In this paper, rice straw was thermochemically modified with citric acid (CA) as esterifying agent. Two introduced free carboxyl groups of esterified rice straw were further loaded with sodium ion to yield potentially biodegradable cationic sorbent. In order to investigate the effect of chemical modification on the cationic dye sorption of rice straw, the removal capacities of native and modified rice straw sorbing a cationic dye (malachite green) from aqueous solution were compared. The effects of various experimental parameters (e.g. initial pH, sorbent dose, dye concentration, contact time) were investigated. For modified rice straw (MRS), the malachite green (MG) removal percentage came up to the maximum value beyond pH 4. For the 250 mg/l of MG solution, the 1.5 g/l or up of MRS could almost completely remove the dye from aqueous solution. Under the condition of 2.0 g/l sorbent used, the percentage of MG sorbed on MRS kept above 93% over a range from 100 to 500 mg/l of MG concentration. The sorption isotherms fitted the Langmuir or Freundlich models. The sorption equilibriums were reached at about 10 h. The sorption processes followed the pseudo-first-order rate kinetics. After chemical modification, the intraparticle diffusion rate constant (k(id)) was obviously increased. The results in this study indicated that MRS was an excellent sorbent for removal of MG from aqueous solution.

Adsorption↗
PubMed1997

Automated quantitation of total protein in cultured skin fibroblasts.

An automated Coomassie Blue method of measuring total protein in cultured skin fibroblasts using a random access analyzer, the Roche MIRA, was compared to a manual dye binding method using bicinchoninic acid. The automated Coomassie Blue method is in common use in many routine laboratories for measurement of total protein in urine and CSF, and was found by us to be significantly faster than a manual protein method, with an average assay time of 2 min per sample. In addition, intra- and inter-run precision were comparable for the automated and manual methods, and compared favourable with other automated methods. We recommend that where the MIRA apparatus is available, consideration be given to its use for protein quantitation of cell or tissue extracts.

Fibroblasts↗
PubMed2001

Influence of resin cement viscosity on microleakage of ceramic inlays.

OBJECTIVES: The aim of the present investigation was to evaluate the effect of the different viscosities of two resin luting cements on microleakage of ceramic inlays at dentinal margins. The effect of the width of the space between inlay and tooth, on the quality of the marginal seal was also investigated. METHODS: Mesial and distal class V cavities were prepared in 48 extracted third molars. The incisal margins of the cavities were in enamel and the cervical margins in dentin. Subsequently, Empress inlays with different cervical margin gap dimensions were fabricated. The mean cervical gap dimensions in the respective groups were as follows: group 1 (27 microm); group 2 (232 microm); group 3 (406 microm). Half the inlays in each group (16) were cemented with a low viscous resin luting cement, and half (16) with a highly viscous resin luting cement. The teeth were subjected to occlusal loading with synchronized thermal cycling in a masticatory simulator. Then, the specimens were immersed in basic fuchsin solution, and dye penetration along the cavity walls was measured. In addition, marginal adaptation was analyzed in the SEM at baseline and after loading, using a replica technique. RESULTS: With regard to dye penetration at dentinal margins, the highly viscous cement performed statistically significantly better at dentin/composite margins than the low viscous cement (p=0.0158). These findings are supported by SEM analysis. SIGNIFICANCE: It is assumed that polymerization stress within the luting cement could not be completely compensated for by larger luting spaces. Highly viscous luting cements are recommended for cementing class V inlays in larger luting spaces.

Aluminum Silicates↗
PubMed1988

A malachite green procedure for orthophosphate determination and its use in alkaline phosphatase-based enzyme immunoassay.

An improved procedure for phosphate determination based on a highly colored complex of phosphomolybdate and malachite green is described. All necessary reagents are combined in one concentrated solution, making the assay sensitive and convenient. The procedure is based on the finding that the dye is easily soluble and stable in the presence of 6 N acid. The addition of Tween 20 is required to stabilize the dye-phosphomolybdate complex at phosphate concentrations above 10 microM. The time of color development at 25 degrees C is about 3 min. The procedure was adopted to measure alkaline phosphate activity in heterogeneous enzyme immunoassay with rho-nitrophenyl phosphate and pyrophosphate as substrates. In both cases, a 4-fold increase in sensitivity in terms of absorbance readings was obtained compared to the standard method based on rho-nitrophenol measurement. In visual analysis, the gain in sensitivity was as high as 20-fold, due to contrast color change (yellow to greenish blue).

Alkaline Phosphatase↗
PubMedDate not supplied

Sentinel node study in early breast cancer.

Since October 1997 60 patients with early breast cancer (T <3 cm) were studied. All patients underwent lymphoscintigraphy with two types of colloid: the first (17 pts) with a particle size <1,000 nm; the second (43 pts) with a particle size <80 nm. The standard procedure consists of injection, on the day before surgery, of 70 MBq of the smaller nanocolloid in 0.4 cc saline divided over four sites, around the lesion or subdermally around the surgical scar. We utilize a low-energy, high-resolution LFOV camera for scintigraphy and a probe specific for the sentinel node during surgery. In 56/60 patients (93.3%) lymphoscintigraphy showed the sentinel node (SN). In two cases the SN was not detected presumably because of lymphatic interruption by an old surgical scar; in the other two cases the sites of injection were too close to the SN, thus masking it. In five cases (9%) the SN was not visualized with the surgical probe but in two of these drainage to the internal mammary chain was observed. The apparently lower sensitivity of intraoperative localization was due to the extra-axillary lymphatic drainage or to the vicinity of the SN to the primary lesion. The SN proved to be metastatic in 12 cases. No false-negative SNs were found. In five cases (10%) the radiolabeled lymph node was the only node containing tumor cells (micrometastases): this result depends on the combined use of hematoxylin-eosin and rapid cytokeratin staining. The application of blue dye was useful for easier identification of the SN but did not allow detection of more SNs. Our preliminary results are extremely encouraging. Considering that at the early stages of breast cancer the likelihood of lymph node metastases is low (20% in our series) and no false negative were reported in this study, we conclude that with SN biopsy axillary lymph node dissection can be avoided, making surgery less aggressive but maintaining accuracy.

Adult↗
PubMed1995

Fluid uptake from the renal medulla into the ascending vasa recta in anaesthetized rats.

1. We have investigated fluid movements between superficial ascending vasa recta (AVR) and the interstitium of exposed papillae of the renal medullae in 15-day-old Sprague-Dawley rats anaesthetized with Hypnorm and Hypnovel. 2. Using a development of the red cell micro-occlusion technique, fluid filtration and reabsorption rates per unit area of vessel wall (Jv/A) were determined in 54 single perfused AVR at known microvascular pressures (Pc). The relation between Jv/A and Pc was non-linear suggesting hydraulic permeabilities (Lp) of 50-100 x 10(-7) cm s-1 cmH2O-1 when Pc was between 0-10 cmH2O and 150-200 x 10(-7) cm s-1 cmH2O-1 when Pc was 10-15 cmH2O. 3. Rates of fluid reabsorption into the AVR estimated by a densitometric technique in a further fourteen vessels were consistent with Lp values of 50-100 x 10(-7) cm s-1 cmH2O-1 when Pc was -2 to 0 cmH2O. 4. The effective oncotic pressures of perfusates containing bovine serum albumin (BSA) were consistent with minimum values for the reflection coefficients of the walls of the AVR to BSA of between 0.59 and 0.72. 5. The concentration of native serum albumin in the papillary interstitial fluid was 9.1 +/- 0.6 mg ml-1 (mean +/- S.E.M., n = 16, from 9 rats), which is approximately 25% of the plasma level. 6. After their microinjection into the medullary interstitium, Patent Blue V and Evans Blue-albumin cleared within 1 min. There was no evidence of preferential movement of either dye towards the base of the exposed renal medulla. 7. Because Lp of the AVR is high, mean pressures of only approximately 3 cmH2O are necessary to account for the total clearance of fluid from the medullary interstitium into the AVR. From published data and from our own observations, it appears that differences in hydrostatic and oncotic pressure across the walls of the AVR are more than sufficient to provide this driving force. The possibility of the clearance of protein from the interstitium into the AVR is discussed.

Absorption↗
PubMed2002

Pulmonary lymphatic mapping in dogs: use of technetium sulfur colloid and isosulfan blue for pulmonary sentinel lymph node mapping in dogs.

Lung cancer is the most frequent cause of cancer death in the United States. The pattern of regional lymph node involvement is a major prognostic factor in a patient with nonsmall cell lung cancer. The accuracy of information obtained about the lymph node status of lung cancer patients can be potentially increased by sentinel node lymphatic mapping. This technique has been well studied in melanoma and breast cancer. It may be useful in increasing the detection of micrometastases and in decreasing the morbidity from complete mediastinal lymphadenectomy. We report an animal pilot study of pulmonary lymphatic mapping. The aim of our study was to gain experience in the surgical techniques for pulmonary sentinel node lymphatic mapping in an animal model prior to its application in humans. Technetium sulfur colloid and isosulfan blue dye were injected into different lobes of the lung followed by attempts to identify the sentinel node draining that specific portion of the lung. Technetium sulfur colloid identified the sentinel node in five of six dogs within 20 min after the radiotracer was injected into the lung parenchyma. Isosulfan blue dye identified the sentinel node in three of six dogs within 5 min. Both the agents are potentially useful, but we found greater technical ease in identifying the sentinel node with technetium sulfur colloid. Two single-institution pilot studies in humans have been performed. A multicentered study to validate and further refine this technique is necessary. Advanced pathologic techniques such as immunohistochemistry and reverse transcriptase-polymerase chain reaction can be used to enhance the accuracy of staging. This may facilitate proper application of novel therapeutic strategies to improve the current dismal prognosis of this disease.

Animals↗
PubMed2003

Decolorization of malachite green and crystal violet by waterborne pathogenic mycobacteria.

Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium scrofulaceum, Mycobacterium marinum, and Mycobacterium chelonae tolerate high concentrations of the dyes malachite green and crystal violet. Cells of strains of those species decolorized (reduced) both malachite green and crystal violet. Because decolorized malachite green lacked antimicrobial activity, the resistance of these mycobacteria could be due, in part, to their ability to decolorize the dyes. Small amounts of malachite green and its reduced, decolorized product were detected in the lipid fraction of M. avium strain A5 cells grown in the presence of malachite green, suggesting that a minor component of resistance could be due to sequestering the dyes in the extensive mycobacterial cell surface lipid. The membrane fraction of M. avium strain A5 had at least a fivefold-higher specific decolorization rate than did the crude extract, suggesting that the decolorization activity is membrane associated. The malachite green-decolorizing activity of the membrane fraction of M. avium strain A5 was abolished by either boiling or proteinase exposure, suggesting that the decolorizing activity was due to a protein. Decolorization activity of membrane fractions was stimulated by ferrous ion and inhibited by dinitrophenol and metyrapone.

Anti-Infective Agents, Local↗
PubMed2004

Adsorptive removal of basic dyes from aqueous phase onto activated carbon of used tea leaves: a kinetic and thermodynamic study.

Activated carbon has been prepared from used tea leaves impregnated with H3PO4 (50%, w/v) and carbonized at 300 degrees C. Its adsorption capacity has been tested for the decolourisation of wastewater containing malachite green and methylene blue. The effect of system variables such as concentration, temperature, pH, contact time, adsorbent dosage and particle size was studied. The optimum pH range is 8-10 and dosage required is 1.2 g/L for 100% removal of both malachite green and methylene blue (100 mg/L). Break-through and exchaustive capacities for malachite green and methylene blue are 300.0 and 314.7 mg/g; 275.0 and 290.4 mg/g, respectively. The adsorption data follow Langmuir model as well as Freundlich model.

Adsorption↗
PubMed1982

Prometaphase banding of human chromosomes with basic fuchsin.

A technique is described for the production of detailed and richly contrasting G-band patterns in human prometaphase chromosomes with the aid of the triphenylmethane dye basic fuchsin. The usefulness of this method is illustrated by its application for the precise analysis of two chromosome 11 rearrangements. It is also demonstrated that high-resolution banding with basic fuchsin can reveal bands not present in the international standard idiogram of human prophase chromosomes (ISCN 1981). The technique described can also be used for easy recognition of the late replicating X chromosome, which stains darker than its early replicating homologue. A preliminary analysis of the late replicating X chromosomes in a 49,XXXXY individual suggests that the three supernumerary X chromosomes do not necessarily replicate synchronously.

Chromosome Banding↗
PubMedDate not supplied

Lymphatic mapping and sentinel lymph node biopsy in the staging of melanoma.

In recent months, we have witnessed a 'paradigm shift' in the management of intermediate-thickness melanoma. The collective experience of the recent past confirms the validity of the 'sentinel' lymph node as being the initial draining site from a specific area of skin. Furthermore, the sentinel lymph node has been confirmed as the most likely site in the regional lymph node basin to harbor occult metastatic disease. Identification of sentinel lymph nodes by visual inspection and intraoperative gamma probe detection after the peritumoral injection of Lymphazurin blue dye and technetium sulfur colloid is a reliable new technique. Staging accuracy also has improved, allowing the precise identification of patients who benefit from avoiding the morbidity of radical lymphadenectomy. The importance of accurate staging has been heightened by data demonstrating effective adjuvant therapy with recombinant interferon-alpha 2B. Precisely defining patient subsets who benefit from adjuvant high-dose interferon-alpha 2B is the current focus of clinical trials designed to maximize the enhanced staging accuracy of the novel approach of sentinel lymph node biopsy.

Biopsy, Needle↗
PubMed1988

Use of a scanning densitometer or an ELISA plate reader for measurement of nanogram amounts of protein in crude extracts from biological tissues.

Protein contents of crude extracts from plant and animal tissues can be rapidly assayed using a Coomassie blue dye-binding procedure combined with scanning densitometry. Total protein is extracted from 100 mg of fresh-frozen or dried-ground tissue using 1 ml of extraction buffer. One-microliter aliquots of standard solutions or crude extracts are spotted in rows on a suitably sized sheet of Whatman 3MM chromatography paper. The dried samples are stained with Coomassie brilliant blue R-250 (0.2%, w/v, in acidified 50% MeOH) for 20 min and rinsed twice with acidified 20% MeOH. After drying, protein concentrations are read as reflectance using a scanning densitometer and peak heights or peak areas recorded using a digital integrator. In an alternative procedure, each spot is cut from the sample sheet and the dye-protein complex eluted in 1% sodium dodecyl sulfate (SDS) using an ultrasonic cleaner. Absorbance is subsequently read in a microwell sample holder at 590 nm with an enzyme-linked immunosorbent assay plate reader. Both procedures offer distinct advantages over previously reported methods. They are significantly faster when large numbers of samples are processed, they avoid interference by chlorophyll, dithiothreitol, SDS, 2-mercaptoethanol, Nonidet P-40, and phenylmethylsulfonyl fluoride (and other protease inhibitors) and they yield marked improvements in sensitivity, providing measurements of protein concentration below 100 and 200 ng.microliter-1, respectively.

Animals↗
PubMed2000

Adhesion testing of a denture base resin with 5 casting alloys.

PURPOSE: This study compared denture base resin shear bond strengths to silicoated Au-Pd, Au-Pd-Ag, Au-Ag-Pd-Cu, high-Pd, and Ni-Cr-Be alloys used to fabricate frameworks for hybrid implant prostheses. Microleakage between alloy and resin was also compared among groups after specimen fracture. MATERIALS AND METHODS: Twelve cylindrical specimens were cast for each alloy. Each specimen was made from a ring-shaped pattern (diameter [d] = 12 mm and height = 4 mm) and machined to achieve uniform hollow centers (d = 6.5 mm). Castings were abraded with 250-micron aluminum oxide and ultrasonically cleaned in distilled water before silicoating. Denture base resin was processed to the internal surfaces of the silicoated specimens. All specimens were thermocycled (1,000 cycles) between 4 degrees C and 50 degrees C, and placed in basic fuchsin dye for a week. A punch (d = 3.8 mm) driven at a cross-head speed of 0.5 mm/min was used to push out the resin specimens. The force required to cause failure was converted to the nominal shear bond strength for each specimen, and mean shear bond strengths for the 5 groups of specimens (N = 12) were compared using one-way analysis of variance and the Tukey-Kramer HSD multiple range test (alpha = 0.05). Six of the 12 debonded resin samples for each alloy were selected at random and evaluated for dye penetration. Using an 80-square grid, the percentage of dye penetration was evaluated with an optical microscope (x25) to determine the percentage of grid area penetrated by the dye. One-way analysis of variance was used to compare the degree of microleakage among groups. RESULTS: The mean resin-alloy shear bond strengths for the Au-Pd (9.6 +/- 3.7 MPa) and Au-Ag-Pd-Cu (9.2 +/- 1.5 MPa) alloys were significantly greater than mean resin-alloy shear bond strength for the Au-Pd-Ag alloy (5.6 +/- 1.9 MPa). No other significant differences in resin-alloy shear bond strengths were noted among the alloy groups. No significant differences were noted for dye penetration into the resin specimens bonded to any of the 5 alloys. The mean grid area penetrated by the dye was 25% when results for the alloys were pooled. CONCLUSIONS: Alloy type influences the shear bond strength of a denture base resin to silicoated alloys, but no difference in bond strength was found between Au-Pd, Au-Ag-Pd-Cu, high-Pd, and Ni-Cr-Be alloys. In addition, under the conditions of this study, all groups showed a similar degree of microleakage, which penetrated approximately 25% of the bonded specimen surface area.

Air Abrasion, Dental↗
PubMed2003

[Lymph node mapping and sentinel node detection in carcinoma of the cervix, endometrium and vulva].

OBJECTIVES: Validity of the sentinel node concept in patients with cervical, endometrial and vulvar cancer. MATERIAL AND METHODS: 47 cases of FIGO stage I and II cervical cancer, 33 cases of first clinical stage of endometrial cancer and 37 patients with FIGO stage I and II of vulvar cancer. In cervical and vulvar cancer preoperative lymphoscyntygraphy and intraoperative lymphatic mapping with blue dye and handheld gamma probe were performed. In patients with endometrial cancer intraoperative lymphatic mapping with blue dye injected into the cervix and into the uterine corpus subserously were done. In the last 10 cases radiolabeled nannocolloid were administered and the patients underwent preoperative lymphoscyntygraphy and intraoperative radio detection of sentinel node. Sentinel nodes were labeled as blue, radioactive, or blue/radioactive. RESULTS: In cervical cancer sensitivity of the dye and radiocolloid methods was 94%, specificity 100% and negative predictive value 97%. Out of 33 cases of endometrial cancer sentinel node was identified in 29 (87.87%) patients. None of women with histological negative sentinel node had metastases in the rest of lymph nodes resected. Sentinel node was detected in all cases of vulvar cancer. The status of sentinel nodes were representative for all lymph node resected. CONCLUSIONS: Concept of sentinel node may be applied first of all for vulvar cancer and also for cervical and endometrial cancer.

Endometrial Neoplasms↗
PubMed1988

Development and evaluation of an automated dye-binding assay for protein in cerebrospinal fluid.

Addition of sodium dodecyl sulfate (SDS) to Coomassie Brilliant Blue reagent equalizes the binding variability of the dye to various proteins and markedly improves the accuracy of quantification of protein in cerebrospinal fluid. In the presence of SDS, the absorption spectrum and the absorption maximum are affected by reaction time and temperature, age of the dye preparation, and protein constituent. I automated this procedure, to optimize precision, enable use of a smaller sample, decrease hands-on time, maintain consistency in the time of reading, and avoid carryover. The results (y) compared well with those of the aca (x), with a Deming de-biased regression equation of y = 0.991x + 14.1 mg/L, Sy.x = 33.4 mg/L. The within-run and between-run precision (CV) was less than 2.5% and less than 4.5%, respectively. Commonly used antibiotics, flucytosine, or amphotericin B do not interfere. This automated procedure is fast and accurate and requires only 10 microL of sample.

Autoanalysis↗
PubMed1975

[Dye-marking and hydrogen clearance in demonstrating acute ischemic myocardial areas].

After acute myocardial infarction (produced by ligation in the dog), the authors marked the borders of infarction with disulphine blue or methylene blue and determined, by means of hydrogen clearance, the blood supply in the centre and at the margin of infarction as well as at different distances from the latter. At the margin of infarction, the value for blood flow amounted to almost 17% of the value measured prior to ligation. Only at a distance of 1 cm. from the visible margin of infarction, 50 to 80% of the initial value were obtained. The extension of the area of infarction beyond the marked visible margin must be considered in surgical interventions.

Animals↗
PubMed1981

Demonstration of axial elements of sex vesicles of spermatocytes using Coomassie brilliant blue.

The axial element of sex chromosomes in the sex vesicle of rat and mouse spermatocytes has been visualized under the light microscope by the dye Coomassie brilliant blue (CBB). After staining in the CBB solution for 3-10 minutes, the axial elements appeared as darkly stained threads in the sex vesicles, whereas in controls stained with Giemsa or carbol fuchsin, the sex vesicles were usually uniformly stained. The axial elements are best seen when chromosome preparations were made by the flame drying technique. In rat spermatocytes the staining quality could be further improved by a brief treatment with trypsin solution (0.025%). The CBB staining procedure is simple and easily controllable. The results suggest that the CBB stained material is protein in nature and is more resistant to trypsin digestion than other nuclear proteins.

Animals↗