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[Medical emergencies following dermatological injections and infusions].

Emergency situations following appropriately administered injections and infusions are an uncommon dermatologic problem. Embolia cutis medicamentosa is an uncommon complication following intramuscular injection which leads to infarct-like cutaneous necrosis. It may develop after gluteal injection of corticosteroids, anti-inflammatory agents and antibiotics and has rarely been described following venous sclerotherapy with polidocanol. Hoigné syndrome is a pseudoanaphylactic or pseudoallergic reaction following intramuscular injection of procaine penicillin, with neuropsychiatric problems developing immediately after the injection. Subacute forms following intramuscular or oral administration of structurally-related antibiotics. The intraoperative use of patent blue dye has a 1-2% risk of allergic reactions. The patient must be monitored carefully following injection for the onset the life-threatening immediate reaction. Extravasation of cytostatic agents is a critical iatrogenic problem in oncology with an incidence of up to 6.5%. Every oncologic department should have an understanding of necrotic risk of the various agents and an emergency kit with instructions for immediate steps, also including agent-specific antidotes as available.

Anaphylaxis↗

Factors affecting failed localisation and false-negative rates of sentinel node biopsy in breast cancer--results of the ALMANAC validation phase.

BACKGROUND: Despite the widespread application of sentinel lymph node biopsy (SLNB) for early stage breast cancer, there is a wide variation in reported test performance characteristics. A major aim of this prospective multicentre validation study was to quantify detection and false-negative rates of SLNB and evaluate factors influencing them. METHODS: Eight-hundred and fourty-two patients with clinically node-negative breast cancer underwent SLNB according to a standardised protocol that used a combination of radiopharmaceutical 99mTc-albumin colloid and Patent Blue V dye. SLNB was followed by standard axillary treatment at the same operation in all patients. RESULTS: Sentinel lymph nodes (SLNs) were identified in 803 (96.1%) of 836 evaluable cases. The median number of SLNs removed per patient was 2 (range 1-9). There were 19 false negatives, resulting in a sensitivity of 263/282 (93.3%) and accuracy 782/803 (97.6%). SLNs were successfully identified by blue dye in 698 (85.6%), by isotope in 698 (85.6%), and by the combination of blue dye and isotope in 782 (96.0%) of 815 patients. Among 276 node positive patients, one or more positive SLNs were identified by blue dye in 251 (90.9%), by isotope in 246 (89.1%) and by the combination of blue dye and gamma probe in 258 (93.5%). Obesity, tumor location other than upper outer quadrant and non-visualisation of SLNs on the pre-operative lymphoscintiscan were significantly associated with failed localisation (p<0.001, p=0.008, p<0.001, respectively). The false-negative rate in patients with grade 3 tumors was 9.6%, compared with 4.7% in those with grade 2 tumors (p=0.022). The false-negative rate in patients who had one SLN harvested was 10.1%, compared with 1.1% in those who had multiple SLNs (three or more) removed (p=0.010). CONCLUSION: SLNB can accurately determine whether axillary metastases are present in patients with early stage breast cancer with clinically negative axillary nodes. Both success and accuracy of SLNB are optimised by the combined use of blue dye and isotope. SLNB success decreases with increasing body mass, tumor location other than the upper outer quadrant and non-visualisation of hot nodes on the pre-operative lymphoscintiscan. This study demonstrates reduction in the predictive value of a negative SLNB in grade 3 tumors.

Adult↗

Quantitative determination of phospholipids using the dyes Victoria blue R and B.

Different phospholipids, except the choline-containing phospholipids phosphatidylcholine, lysophosphatidylcholine, and sphingomyelin, formed complexes with the dye Victoria blue R, which selectively partitioned into the chloroform phase of chloroform/ethylene glycol/glycerol biphasic solvent system, and were quantitatively estimated at 590 nm. Considerable amounts of water, alcohols, nonlipid phosphates, neutral lipids, free fatty acids, and some detergents did not interfere with the formation of phospholipid-dye complexes. This special advantage of the method described allowed combined phospholipid extraction and estimation procedures in one test tube. Because of its high sensitivity (about 24.00 OD units/mumol of phosphatidic acid and about 10.25 OD units/mumol of other phospholipids), specificity, and simplicity, the proposed phospholipid assay appears to be very useful for rapid analyses of lipid extracts as well as TLC spots or suspensions of biological materials, as demonstrated for membranes and cells of Micrococcus lysodeikticus. The applicability of the dye Victoria blue B to the quantitative determination of phospholipids, except phosphatidylcholine, lysophosphatidylcholine, and sphingomyelin, at 605 nm using chloroform/ethylene glycol/glycerol/water and pentane (hexane)/ethyl acetate/isopropanol/water biphasic solvent systems with similar sensitivities and of sodium dodecyl sulfate in the pentane-containing system with high sensitivity (22.96 OD units/mumol) is also shown. The adaptation of this phospholipid assay to the determination of phospholipases C and D and to the differential quantitation of choline-containing phospholipids using additional phospholipid estimation techniques is discussed.

Chloroform↗

An assay for heparin by decrease in color yield (DECOY) of a protein-dye-binding reaction.

The interference by heparin and some related molecules with the well-known Bradford dye-binding assay for proteins is used as the basis of a rapid, sensitive method for the quantitation of these polysaccharides. Whereas the available methods for the assay of glycosaminoglycans have lacked specificity for sulfated polyanions in general and for heparin in particular, the procedure described here distinguishes among different uronic acid derivatives and, when performed in conjunction with heparinase digestion or cetylpyridinium chloride precipitation, can be used to determine heparin content of complex biological samples.

Animals↗

Effects of sodium dodecylsulphate, dye concentration and paraprotein on coomassie blue dye-binding assays for protein in urine.

Various Coomassie Blue reagents, containing either increased dye concentration or added sodium dodecylsulphate, were compared with a biuret method for the assay of total protein in urine. When immunoglobulin free light chain protein or immunoglobulin paraprotein were present, results from the Coomassie Blue methods were up to 50% lower than with the biuret method; increased dye concentration did not improve comparability substantially, but the addition of sodium dodecylsulphate reduced the bias to about 20%. When neither free light chain protein nor immunoglobulin paraprotein was present, results from the Coomassie Blue methods were only about 30% lower. The addition of sodium dodecylsulphate reduced this bias to 10%. Correlations between the biuret and the Coomassie Blue method were best when the Coomassie Blue reagent contained 40 mg/L sodium dodecylsulphate (r better than 0.98 in all groups; p less than 0.001).

Binding, Competitive↗

Differential staining of two subpopulations of Purkinje neurons in rat cerebellum with acid dyes.

We present a new method that stains differently two subpopulations of Purkinje cells in the adult rat. Deparaffinized sections of cerebella, fixed by perfusion with buffered glutaraldehyde or Bouin's fluid were stained with 0.5% light green in 50% ethanol (10-30 min). The excess dye was removed with saturated aqueous picric acid (10-30 min). At this point some Purkinje cells appeared as lightly stained neurons, while others were strongly stained. Slides were immersed in 0.5% aqueous acid fuchsin for approximately 1 min until the lightly stained neurons acquired a red color. Following immersion in 1% phosphotungstic acid, slides were rapidly dehydrated in ethanol, passed to xylene and mounted in Canada balsam. Two subpopulations of Purkinje cells differing in their protein content in somata and proximal dendrites stained differentially by this method. They occurred in all coronal and sagittal sections and in patches or stripes. Their relative proportion varied from lobule to lobule. A second staining method used potassium permanganate as the sole staining reagent. The staining reagent can be used on sections previously stained with the acid dyes. Purkinje cells appeared as subsets of brownish to deep brown stained neurons, the latter ones corresponding to green stained cells in the dichromic method. The results obtained indicated that the subpopulations reflect real differences among individual neurons and are not artifacts. The technique holds promise for identifying and localizing sub-sets of Purkinje cells differing in their protein content under normal and experimental conditions and for their further characterization by combined staining and histochemical procedures.

Animals↗

Selective destruction of protein function by chromophore-assisted laser inactivation.

Chromophore-assisted laser inactivation of protein function has been achieved. After a protein binds a specific ligand or antibody conjugated with malachite green (C.I. 42,000), it is selectively inactivated by laser irradiation at a wavelength of light absorbed by the dye but not significantly absorbed by cellular components. Ligand-bound proteins in solution and on the surfaces of cells can be denatured without other proteins in the same samples being affected. Chromophore-assisted laser inactivation can be used to study cell surface phenomena by inactivating the functions of single proteins on living cells, a molecular extension of cellular laser ablation. It has an advantage over genetics and the use of specific inhibitors in that the protein function of a single cell within the organism can be inactivated by focusing the laser beam.

Chemical Phenomena↗

The marginal seal of a flowable composite, an injectable resin modified glass ionomer and a compomer in primary molars--an in vitro study.

The present study was carried out to compare the marginal microleakage of some newer materials viz. a flowable composite, an injectable resin modified glass-ionomer and a compomer in Class I cavities of 30 non carious primary molars. After 0.5% basic fuchsin dye penetration and sectioning, the teeth were studied under stereomicroscope. The results obtained revealed that flowable composite showed significantly lower microleakage (p<0.05) as compared to injectable resin-modified glass ionomer and compomer. However, no significant difference was observed when injectable resin modified glass-ionomer cement was compared to compomer. This concludes that flowable composite materials adhere better to the primary teeth than resin modified glass ionomer and compomer.

Adhesiveness↗

[Preparation of nanometer TiO2 doped with upconversion luminescence agent and investigation on degradation of ethyl violet using visible light].

A novel upconversion luminescence agent 40CdF2 x 60BaF2 x 1.6Er2O3 was synthesized and the fluorescent spectrum was determined. This upconversion luminescence agent can emit five upconversion fluorescent peaks whose wavelengths are all below 387nm under the excitation of 488 nm visible light. This upconversion luminescence agent was mixed into nanometer TiO2 powder by ultrasonic dispersion and the doped nanometer TiO2 photocatalyst utilizing visible light was prepared. The doped TiO2 powder was charactered by XRD and TEM and its photocatalytic activity was checked through the photocatalytic degradation of ethyl violet dye as a model compound under the visible light irradiation emitted by three basic color lamp. Otherwise, in order to compare the photocatalytic activities the same experiment was carried out for undoped photocatalytic TiO2 powder. The degradation ratio of ethyl violet dye in the presence of doped nanometer TiO2 powder reached 99.68% under visible light irradiation at 12.0 h which was obviously higher than the corresponding degradation ratio in the presence of undoped nanometer TiO2 powder, which indicate that the upconversion luminescence agent prepared as dopant can effectively turn visible lights to ultraviolet lights which are absorbed by nanometer TiO2 particles and produce the electron-cavity pairs.

Catalysis↗

Chemical safety of U.S. Navy Fleet soda lime.

Contamination was suspected of U.S. Navy Fleet soda lime (High Performance Sodasorb) when an ammonia-like odor was reported during its use in August 1992. This material contained indicator dye and was used for carbon dioxide absorption during diving. This incident had a major impact on the U.S Navy diving program when the Navy temporarily banned use of Sodasorb and authorized Sofnolime as an interim replacement. The Naval Medical Research Institute was assigned to investigate. Testing involved sampling from the headspace (gas space) inside closed buckets and from an apparatus simulating conditions during operational diving. Volatile organic compounds were analyzed by gas chromatography and mass spectrometry; ammonia and amines were measured by infrared spectroscopy. Significant amounts of ammonia (up to 30 ppm), ethyl and diethyl amines (up to several ppm), and various aliphatic hydrocarbons (up to 60 ppm) were detected during testing of both Sodasorb and Sofnolime. Contaminants were slowly removed by gas flow and did not return. The source(s) of the ammonia and amines are unknown, although they may result from the breakdown of the indicator dye. Hydrocarbon contamination seems to result from the materials of which the bucket is constructed. Unfortunately, evaluation of potential hazards associated with this contamination is difficult, due in part to the large number of variables of operational use and the absence of appropriate exposure limits. Based on these findings, the U.S. Navy has begun to phase in, for all diving, non-indicating soda lime that will be required to meet defined contaminant limits.

Amines↗

Comparison of intradermal and subcutaneous injections in lymphatic mapping.

BACKGROUND: Sentinel node biopsy (SNB) for melanoma, with its intradermal (ID) injection, has a higher success rate than SNB for breast cancer, which is typically performed with a subcutaneous (SC) or peritumor injection. It is hypothesized that this is in part due to a slower transit time of lymphatic mapping agents through the parenchymal lymphatics of the breast. No study has investigated differences in transit time between different tissues to account for this clinical observation. The goal of the study was to compare transit time between ID and SC injections with common agents used in lymphatic mapping. METHODS: Four injection sites on five domestic pigs were used. Sites were bilateral and included cervical, forelimb, hindlimb, and flank areas. Agents included technetium sulfur colloid (Tc99, filtered and unfiltered), isosulfan blue (IB) dye, and fluorescein (FL) dye. At each site both ID and SC injections were made and the transit time to reach the sentinel node was recorded. The transit time differences were calculated per centimeter distance from the draining lymph node basin. RESULTS: Sentinel nodes were identified draining all sites and found to be hot, blue, or fluorescent (using a Wood's lamp for identification). The cervical and forelimb injection sites drained to the same cervical lymph node basin and both SC and ID injection sites drained to the same sentinel node. Similarly, the hindlimb and flank injection sites both drained to inguinal lymph node basins. The slowest transit time occurred with Tc99 injected SC and the fastest occurred with Tc99 injected ID, whereas both FL dye and IB traveled rapidly to the sentinel node whether injected SC or ID. Large differences were found using unfiltered Tc99 depending on its injection ID (2.7 s/cm +/- 0.5) vs SC (249 s/cm +/- 14.7, P = 0.008). CONCLUSIONS: Tc99 ID injections were significantly faster than SC injection. The slowest and fastest SC injection agents were unfiltered Tc99 and IB, respectively. Dermal injections provide faster transit of lymphatic agents and may improve the identification rate when applied to patients with breast cancer.

Animals↗

Two sulphonated dye compounds which compete for inositol 1,4, 5-trisphosphate binding to rat liver microsomes: effects on 5'-phosphatase activity.

The ability of heparin to interact with the Ins 1,4,5-P3 receptor is dependent on its chain length and degree of sulphation. Here we report results obtained with two sulphonated dye compounds of known structures and molecular weights below 1000, cibacron blue and Patent blue. Both compounds compete for Ins 1,4,5-P3 binding to rat liver microsomes and also inhibit Ins 1,4,5-P3 5'-phosphatase activity in the same preparation. Comparison with the effects of heparin show these to be two separate actions of the compounds.

Animals↗

Use of a compomer in Class V restoration: a microleakage study.

OBJECTIVE: The purpose of this investigation was to evaluate the microleakage of Class V lesions restored with a compomer material (Dyractflow) using 2 single-bottle bonding systems. METHOD AND MATERIALS: Sixty-four Class V cavity preparations were made on 32 sound, extracted human premolars, 1 preparation on the facial surface and 1 on the lingual surface. Teeth were then randomly divided into 2 groups. Group 1, which served as the control group, was restored as follows: group 1a, facial surface with PQ1 single-bottle bonding system plus TPH resin composite; group 1b, lingual surface with Prime & Bond NT single-bottle bonding system plus TPH. Group 2 was restored as follows: group 2a, facial surface with PQ1 plus Dyractflow compomer material; group 2b, lingual surface with Prime & Bond NT plus Dyractflow. After 24 hours of storage in water and subsequent thermocycling, each specimen was immersed in 0.2% basic fuchsin dye for 24 hours. The teeth were sectioned longitudinally and horizontally, and dye penetration at enamel and cementum margins was viewed at x30 magnification by 2 independently calibrated evaluators. Specimens were evaluated as either having leaked or not having leaked. RESULTS: No statistically significant differences in microleakage were noted between group 1 and group 2 specimens. CONCLUSION: Both single-bottle bonding systems, PQ1 and Prime & Bond NT, demonstrated equal effectiveness in reducing microleakage beneath compomer and composite restorations.

Bisphenol A-Glycidyl Methacrylate↗

Effects of patent blue on human retinal function.

BACKGROUND: Dye solutions for intraoperative staining of epiretinal membranes and the internal limiting membrane improve the visualisation of these thin structures and facilitate their removal. Lately, the application of patent blue for staining of the internal limiting membrane has been proposed as an alternative to the standard procedures during macular hole surgery. In the present study we investigated the effects of patent blue 0.48% on human retinal function. METHODS: An isolated human retina preparation was perfused with a standard solution and the electroretinogram (ERG) was recorded repeatedly. After recording of stable ERG amplitudes the nutrient solution was substituted by a commercially available patent blue solution. The duration of retinal exposure to the dye solution was varied between 15 s and 4 min. Thereupon, the preparation was reperfused with standard solution for 2 h. The percentage of b-wave reduction after exposition was calculated. RESULTS: No effects on the human ERG were seen after 15 and 30 s of dye application. Reversible reductions of the b-wave amplitude were found for an exposure period of 60 and 120 s, respectively. After 4 min of patent blue application a persistant b-wave amplitude reduction by 40% was found. CONCLUSIONS: Patent blue affects human retinal function when applied for at least 1 min. However, no irreversible effects on the human ERG were seen even after 2 min of retinal exposure to patent blue. Thus, toxic effects on retinal function after intraoperative short-term application of patent blue 0.48% appear unlikely to occur.

Coloring Agents↗

Quantitation of proteins on Coomassie blue-stained polyacrylamide gels based on spectrophotometric determination of electroeluted dye.

A procedure for quantitating proteins on Coomassie blue-stained polyacrylamide gels is presented. The method is based on the observations that the dye is rapidly eluted electrophoretically from stained protein bands or spots in the presence of sodium dodecyl sulfate, and that the eluted dye is nondialyzable. Protein may therefore be assayed indirectly by measuring the dye in electroeluents spectrophotometrically. Moreover, the stain elutes more rapidly than the protein, allowing separate recovery of the protein for further analysis. The assay is independent of band or spot size, and does not involve physical disruption of the gel piece or any chemical treatment harsher than the staining process itself. The technique has been applied to the contractile proteins myosin, actin, and commercially obtained standards resolved by one-dimensional electrophoresis, and to proteins in nuclear extracts of HeLa cells on two-dimensional gels.

Animals↗

Isosulfan blue causes factitious methemoglobinemia in an infant.

A 9 month-old girl was given subcutaneous isosulfan blue to outline lymphatic channels during surgery for thoracic duct ligation. Her pulse oximetry values rapidly declined to a nadir of 85%, 35 min after dye injection. Arterial blood gases revealed methemoglobinemia ([MetHb] = 6.5%). Although abnormal pulse oximetry has already been reported in association with isosulfan blue, methemoglobinemia has not previously been reported. The absorption spectrum for isosulfan blue was determined and when superimposed on that of methemoglobin it was found to have an overlying peak. Interference by the dye was postulated to have caused the abnormal methemoglobin result. The phenomenon was simulated in vitro by adding isosulfan blue to whole blood, and analysing it in the same blood gas analyser as was used for the case, as well as another for comparison. One blood gas analyser reported elevated methemoglobin concentration and the other did not. The samples were sent to a reference laboratory using a chemical method to detect methemoglobin to confirm that the elevated methemoglobin level was spurious.

Coloring Agents↗

Light microscopic distinction between elastin, pseudo-elastica (type III collagen?) AND INTERSTITIAL COLLAGEN.

Distinction between elastin and collagen in arteriosclerotic lesions is difficult because the so-called elastica stains are bound also by collagen fibers which resemble collagen of premature infants. Investigations of effects of organic solvents on dye binding led to the development of methods for selective demonstration of pseudo-elastica, and for simultaneous visualization of elastin and pseudo-elastica in contrasting colors. Paraffin sections of human autopsy material were stained with solutions of resorcin-fuchsin, orcein or aldehyde fuchsin in absolute ethanol. In other series, sections pretreated with this resorcin-fuchsin solution were counter-stained with tannic acid-phosphomolybdic acid (TP)-dye technics. Solutions of these "elastica stains" in absolute ethanol colored only pseudo-elastica; elastin, e.g. elastic membranes of aorta, remained unstained. In sections counterstained with TP-dye technics elastin was colored red; pseudo-elastica retained the purplish blue coloration imparted by resorcin-fuchsin. Other collagens were stained yellow. A review of the literature showed that until the 1920's elastin was classified as a gelatinoid of the collagen group. Elastic fibers were identified by mechanical properties, not a particular chemical composition. Hence, the elastic fibers of classical histology cannot be equated with the elastin of modern chemistry. Correlation of histochemical observations with chemical data indicates that the collagenous pseudo-elastica corresponds to [alpha1(III)]3 collagen.

Adult↗

Total protein measurement using a fiber-optic evanescent wave-based biosensor.

A novel method and instrumental system to determine the total protein concentration in a liquid sample is described. It uses a fiber optic total protein sensor (FOPS) based on the principles of fiber optic evanescent wave spectroscopy. The FOPS applies a dye-immobilized porous glass coating on a multi-mode optical fiber. The evanescent waves at the fiber optic core-cladding interface are used to monitor the protein-induced changes in the sensor element. The FOPS offers a single-step method for quantifying protein concentrations without destroying the sample. The response time and reusability of the FOPS are evaluated. This unique sensing method presents a sensitive and accurate platform for the quantification of protein.

Biosensing Techniques↗